Embryonic and larval development of a hybrid between kelp grouper Epinephelus moara ♀ × giant grouper E. lanceolatus ♂ using cryopreserved sperm

Hybridization was used to take advantage of desirable traits in offspring. In the present study, we applied the cryopreserved Epinephelus lanceolatus sperm into interspecific hybridization with E. moara. Successful hybridization between these two species was achieved and cultured in 23–24°C seawater (34‰). There was no difference in survival rate between hybrid (E. moara ♀ × E. lanceolatus ♂) and non‐hybrid (E. moara ♀ × E. moara ♂) at 24 hrs post hatch (HPH), but less hybrid (14.35 ± 8.02%) hatched than non‐hybrid (93.60 ± 1.65%), which might be due to irregularity cell cleavage and skeletal deformities from the formation of embryonic body to the later embryonic development. Similar phenomenon was found in hybrid embryos from fertilization with fresh sperm, indicating that species variations between parents, rather than cryopreserved sperm, resulted in deformities in embryos. Mean hatch time of the hybrid was 1 hr faster than that of E. moara. The hybrid was 1.95 ± 0.06 mm in total length when newly hatched and reached 38.00 mm at 58 days post hatch (DPH), which showed faster growth than E. moara (recorded in the previous study). Considering its faster growth, E. moara × E. lanceolatus hybrid was a potential breeding production in aquaculture. Over 212,000,000 larvae have been produced and launched in the market since 2015. The results of this study also shed some lights on further comparative studies in grouper hybrid performance.     

Survival and growth of geoduck clam larvae (Panopea generosa) in flow‐through culture tanks under laboratory conditions

The geoduck clam, Panopea generosa, is a species from the west coast of Baja California, Mexico, and the optimization of seed production systems is still a limiting factor for its aquaculture. In this study, a flow‐through culture system was designed and tested in P. generosa larvae. Survival and growth was compared in triplicate 45‐L fiberglass tanks using three larval densities (5, 10 and 15 larvae/ml). A head tank kept constant the water inflow, with a daily renewal rate of 1.8× tank volume. The food (Isochrysis spp.) was dosed according to the ingestion rate of larvae and the dilution rate. Survival decreased linearly during the first 10 days and reached asymptotic values of ca. 20% (15 larvae/ml) and 50% (densities of 5 and 10 larvae/ml) afterwards. Mean shell length at the end of the experiment (243 ± 1.8 to 270 ± 0.7 μm) was not statistically different among treatments, even though a trend towards higher gross growth rate was observed in the treatment with the lowest density (9.5 μm/day) relative to the rest of the treatments (8.5 μm/day). It is concluded that P. generosa larvae can be successfully grown in flow‐through systems at maximum densities of 10 larvae/ml without significantly affecting their survival and growth rates. The system design was reliable, kept a constant water flow with reduced maintenance, and may represent an important option in the laboratory for increasing the stocking density of Panopea species during the larval phase.     

Water pH and Prochilodus lineatus larvae survival

The present study aimed to determine the influence of water pH on survival of curimbatá, Prochilodus lineatus, larvae. Forty-five 2-l glass aquaria with artificial aeration and no water exchange were stocked at a density of 10 larvae l⁻¹. The pH values used as treatments were 3.7 ± 0.0, 4.0 ± 0.0, 4.4 ± 0.2, 4.6 ± 0.2, 4.8 ± 0.2, 5.1 ± 0.2, 5.3 ± 0.2, 5.6 ± 0.2, 7.2 ± 0.2, 8.7 ± 0.4, 9.0 ± 0.4, 9.2 ± 0.4, 9.4 ± 0.6, 9.7 ± 0.5 and 10.0 ± 0.5 in a completely random experimental design with three repetitions. Water pH was maintained by the addition of NaOH or H₂SO₄ solutions. After 72 h of experiment, no survival was registered at pH below 4.6, 1.5% survival at pH 4.6, and about 50% survival at pH 9.4 and above. Survival rates between 70 and 80% were registered at pH 4.8–5.6 and at pH 7.2, whereas over 90% survival was registered at pH between 8.7 and 9.2.     

Effects of different natural or prepared diets on growth and survival of juvenile spider crabs, <Emphasis Type="Italic">Maja brachydactyla</Emphasis> (Balss, 1922)

The effects of different diets (natural or pellets) on growth, survival, and moulting interval of juvenile spider crabs, weighing between 0.011–1.56 g and up to 17.6 mm in carapace length, were tested over a period of 90 days. During experiment I, five diets were tested: (1) frozen shrimp—Paleomonetes sp., (2) fresh mussels—Mytilus sp., (3) white fish fillets—Merlucius merlucius, (4) blue fish fillets—Sardina pilchardus, and (5) commercial crustacean pellets. Spider crabs fed fresh mussels grew larger (0.98 ± 0.69 g) and had higher growth rates (4.0 ± 0.7 %BWd⁻¹) compared to the other four diets. The crabs fed shrimp pellet and frozen shrimp grew to intermediate sizes and were smaller than the ones fed fresh mussels, but they were larger than spider crabs fed either blue or white fish fillets (0.46 ± 0.63 and 0.26 ± 0.13 g, respectively) compared to the ones fed white fish fillets (0.12 ± 0.04) and blue fish fillets (0.04 ± 0.02 g). The spider crabs fed blue fish fillets only lasted until day 60 of the experiment, after this day none of the 20 fed this diet were left. During experiment II, two diets were tested: (1) white and blue fish fillets and (2) commercial fish pellet. There were no differences in growth both in weight or carapace length (2.9 ± 1.8 and 2.1 ± 1.5 g in weight, and 18.9 ± 5.0 and 17.7 ± 3.3 mm, respectively) at the end of the experiment. Similarly, there were no differences in growth rates in weight between the two diets (1.2 ± 0.4 and 0.9 ± 0.3 %BWd⁻¹, respectively) or in carapace length (0.4 ± 0.1 and 0.4 ± 0.2 %BWd⁻¹, respectively). Fresh mussel appears to be a very good diet to culture the early stages of this species, while shrimp pellets also deliver acceptable results. On the contrary, frozen shrimp, fish fillets either from blue or white species (much higher lipid content in the blue species), and fish pellets were found to be bad diets for the culture of the early stages of M. brachydactyla.     

Optimization of growth requirements and scale‐up cultivation of freshwater algae Desmodesmus armatus using response surface methodology

In order to optimize the optimal cultivation conditions of freshwater algae Desmodesmus armatus for biomass production. In this study, the optimum source of carbon, nitrogen and intensity of light were investigated. Particularly, the variables which are affect the biomass of D. armatus was screened by the Plackett‐Burman (PB) method. Furthermore, the optimized medium composition using central composite design (CCD) of response surface method (RSM) central for D. armatus was reconstituted accordingly to have 0.93 g/L nitrate, 0.04 g/L phosphate, 0.15 g/L magnesium sulphate and 0.07 g/L bicarbonate, and optimum growth conditions of temperature at 27°C, light intensity of 108 μmol m^?2 s^?1, pH 7.00 and air flow of 0.50 L/min. After 12 days, the biomass, protein and polysaccharose content were 1.65 ± 0.15 g/L, 53.61 ± 1.25% and 6.15 ± 0.43%, respectively. Finally, the optimized conditions were applied to the outdoor 800‐L photobioreactor for scale‐up cultivation.     

Differences in uptake and killing of pathogenic and non‐pathogenic bacteria by haemocyte subpopulations of penaeid shrimp,Litopenaeus vannamei, (Boone)

Phagocytosis is an important function of both invertebrate and vertebrate blood cells. In this study, the phagocytic activity of haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei, (Boone), against pathogenic and non‐pathogenic particles was investigated in vitro. The haemocytes of penaeid shrimp were firstly separated by centrifugation on a continuous density gradient of iodixanol into four fractions with five subpopulations (sub), of which sub 1 (hyalinocytes) and sub 4 (semi‐granulocytes) have the main function in phagocytosis of both pathogenic and non‐pathogenic bacteria as well as fluorescent polystyrene beads. It was found that these haemocyte subpopulations engulfed virulent Vibrio campbellii and Vibrio harveyi at a higher rate than non‐virulent Escherichia coli and polystyrene beads. When these bacteria were mixed with shrimp haemocyte subpopulations and incubated for 180 min, the percentage of viable intracellular V. campbellii (25.5 ± 6.0%) recovered was significantly higher than the percentage recovered from V. harveyi (13.5 ± 1.1%). No viable intracellular E. coli was observed in this study. In contrast to V. harveyi and E. coli, V. campbellii containing endosomes did not acidify in time. Incubation of haemocyte subpopulations with the most virulent V. campbellii strain resulted in a significant drop in haemocyte viability (41.4 ± 6.3% in sub 1 and 30.2 ± 15.1% in sub 4) after 180 min post‐inoculation in comparison with the less virulent V. harveyi (84.1 ± 5.6% in sub 1 and 83.4 ± 4.1% in sub 4) and non‐virulent E. coli (92.7 ± 2.8% in sub 1 and 92.3 ± 5.6% in sub 4) and polystyrene beads (91.9 ± 1.6% in sub 1 and 84.4 ± 3.4% in sub 4). These findings may be a valuable tool for monitoring shrimp health and immunological studies.     

Spawning and rearing of Holothuria tubulosa: A new candidate for aquaculture in the Mediterranean region

Holothuria tubulosa (Gmelin, 1788) has recently shown an increased demand in Asian markets, becoming one of the intensively exploited holothurian species in the Mediterranean Sea. A risk is that over‐harvesting is likely affecting both the species' natural stocks and the benthic communities. In this scenario, sea ranching and restocking through aquaculture could assist in mitigating its overexploitation. This study is the first to demonstrate the successful artificial breeding and rearing of H. tubulosa, and its consequent potential as a new species for the Mediterranean aquaculture industry. Here we describe the spawning induction, larval development and early juvenile growth in hatchery cultures, aimed at developing a spawning and rearing protocol for this species. The trials were conducted from July to October in both 2014 and 2015. Holothuria tubulosa was induced to spawn by testing four different methods. Thermal stimulation plus thermal shock emerged as the most efficient method to obtain active and healthy gametes. Larval development in H. tubulosa progressed through five stages, reaching the juvenile stage in 27 days. Two different microalgal feeding regimens were tested for larval breeding. Under the best feeding conditions, 7% of the larvae metamorphosed into settled juveniles, adhering to artificial substrates previously conditioned with benthic biofilm. Our results indicate that H. tubulosa shows good performance in hatchery rearing during the larval phases, indicating that this species could be a new candidate for aquaculture in the Mediterranean region, both for production and restocking proposes.     

Physicochemical and kinetic characteristics of rhodanese from the liver of African catfish <Emphasis Type="Italic">Clarias gariepinus</Emphasis> Burchell in Asejire lake

Two forms of rhodanese were purified from the liver of Clarias gariepinus Burchell, designated catfish rhodanese I (cRHD I) and rhodanese II (cRHD II), by ion-exchange chromatography on a CM-Sepharose CL-6B column and gel filtration through a Sephadex G-75 column. The apparent molecular weight obtained for cRHD I and cRHD II was 34,500 ± 707 and 36,800 ± 283 Da, respectively. The subunit molecular weight determined by sodium dodecyl sulphate–polyacrylamide gel electrophoresis was 33,200 ± 283 and 35,100 ± 141 Da for cRHD I and cRHD II, respectively. Atomic absorption spectrophotometric analysis revealed that cRHD II contained a high level of iron (Fe), which presumably was responsible for the brownish colour of the preparation. In contrast, no Fe was identified in cRHD I, and its preparation was colourless. Further characterization of cRHD II gave true Michaelis–Menten constant (K _m) values of 25.40 ± 1.70 and 18.60 ± 1.68 mM for KCN and Na₂S₂O₃, respectively, an optimum pH of 6.5 and an optimum temperature of 40°C. The Arrhenius plot of the effects of temperature on the reaction rate consisted of two linear segments with a break occurring at 40°C. The apparent activation energy values from these slopes were 7.3 and 72.9 kcal/mol. Inhibition studies on the cRHD II enzyme showed that the activity of the enzyme was not affected by Mn²⁺, Co²⁺, Sn²⁺, Ni²⁺ and NH₄ ⁺, but Zn²⁺ inhibited the enzyme considerably.     

Identification of facultative anaerobic bacteria isolated from the intestine of the minke whale <Emphasis Type="Italic">Balaenoptera acutorostrata</Emphasis> by 16S rRNA sequencing analysis

There are few reports in the literature about the isolation of bacteria from whale intestine. In this report, we counted colony-forming units in the feces obtained from three female common minke whales (Balaenoptera acutorostrata). The number of colony-forming units ranged from (2.2 ± 0.4) × 10⁵ to (8.9 ± 2.0) × 10⁸ per gram (wet weight) of excrement. 16S rRNA gene sequences of 141 isolates were determined. These strains were identified as Enterococcus faecalis, Enterococcus sp., Enterobacter cloacae, Enterobacter sp., Escherichia coli, Edwardsiella ictaluri or Clostridium sp. The data suggested that the facultative anaerobic population of the intestinal bacterial flora of the minke whale was similar to that of ground mammals.     

Pelagic larval duration of three amphidromous Sicydiinae gobies (Teleostei: Gobioidei) including widespread and endemic species

Sicydiinae species have an amphidromous life cycle during which they undergo a pelagic larval phase allowing them to disperse through the ocean and to recruit in distant island rivers. Hypotheses for the differences observed in dispersal abilities between species include the variation in pelagic larval duration (PLD). However, the implication of the PLD as a proxy for explaining the dispersal ability of a species is not clear in the Sicydiinae subfamily. In this study, otolith microstructure of three Sicydiinae species was analysed. One of these species, Sicyopus zosterophorum, has a widespread distribution in the West Pacific area, whereas the other two species, Smilosicyopus chloe and Akihito vanuatu, are endemic to New Caledonia and to Vanuatu, respectively. Deposition of the daily growth increments on the otoliths of S. zosterophorum was validated using an alizarin complexone time marking technique. We estimated the PLD for the three species by counting the number of growth increments from the core to the metamorphosis check mark, and it was shorter than the one of previous studies on Sicydiinae species. The PLD of the widespread species, S. zosterophorum (54.6 ± 5.6 days), was similar to those of the endemic species, S. chloe (53.6 ± 5.7 days) and A. vanuatu (55.4 ± 7.5 days). Here, we show that in contrast to the most diverse Sicydiinae genus, Sicyopterus, the PLD could not explain endemism, and we must take into account other elements to explain the differences observed in the distribution range.     

Effects of mono‐species and bi‐species microalgal diets on the growth,survival and metamorphosis of auricularia larvae of the California sea cucumber,Parastichopus californicus (Stimpson, 1857)

The effects of mono‐species and bi‐species microalgal diets on body length, survival and metamorphosis of auricularia larvae of the California sea cucumber, Parastichopus californicus, were tested in two separate laboratory feeding experiments. The first examined eight single species of microalgae – Chaetoceros calcitrans (Cc), Chaetoceros muelleri (Cm), Dunaliella tertiolecta (Dt), Isochrysis sp. (Tahitian strain: TISO) (Ti), Pavlova lutheri (Pl), Phaeodactylum tricornutum (Pt), Tetraselmis suecica (Ts) and Thallassiosira pseudonana (Tp). The second experiment took the five most effective single species from the first experiment (i.e. Cc, Cm, Dt, Pl and Ti) and presented them in all possible binary combinations. In the first experiment, Cc was determined to be the best diet on the basis of generating a significantly higher metamorphic rate (65.5 ± 4.2%, mean ± SD) than any other species and leading to one of the best survival rates (72.3 ± 7.6%) and greatest body lengths (831.6 ± 5.6 μm at day 14). In the second experiment, diets containing Cc generally produced the highest survival and metamorphic rates and greatest body lengths (except Cc/Cm) of all mixed diets, although no bi‐species treatment was ever significantly superior to a control of Cc alone. Of the various mono‐species and bi‐species diets evaluated for rearing auricularia larvae of P. californicus, Cc (either alone or mixed with other species) is recommended.     

The use of alternative prey (crayfish, <Emphasis Type="Italic">Procambarus clarki</Emphasis>, and hake, <Emphasis Type="Italic">Merlucius gayi</Emphasis>) to culture <Emphasis Type="Italic">Octopus vulgaris</Emphasis> (Cuvier 1797)

The effects of two alternative prey (crayfish and hake) were tested on growth and survival of both juveniles and adults of Octopus vulgaris in two experiments. Octopuses fed the control (squid) were larger (3.0 ± 0.7 kg) than those fed crayfish (2.4 ± 0.6 kg) at the end of experiment I. Similarly, overall growth rates were higher for octopuses fed squid (1.7 ± 0.3 and 1.2 ± 0.2 %BW day⁻¹, respectively). Average feeding rates for the experiment were not different, being 6.5 ± 0.9 and 7.5 ± 0.9 %BW day⁻¹, respectively, for octopuses fed either squid or crayfish. Nevertheless, food conversions for the experiment were higher (42.4 ± 2.7%) for octopuses fed squid compared to the ones fed crayfish (23.9 ± 1.9 g). For experiment II, hake and crayfish were compared to squid; the final weight of octopuses fed squid, hake or crayfish was 1,183.0 ± 242.7 g, 1,175.6 ± 240.1 g and 922.3 ± 160.1 g, respectively. Overall growth rates for the experiment were 1.9 ± 0.2 %BW day⁻¹, 1.9 ± 0.3 %BW day⁻¹ and 1.1 ± 0.3 g, respectively. Final weight and growth rates were never different (P > 0.05) between octopuses fed squid and hake, but were always higher (P < 0.05) compared to the ones fed crayfish. Average feeding rates for experiment II were similar for the three diets, and of 4.6 ± 1.5, 4.2 ± 1.3 %BW day⁻¹ and 5.1 ± 0.9 %BW day⁻¹, respectively, for octopuses fed squid, hake or crayfish. Food conversions for experiment II were of 41.0 ± 9.6%, 40.5 ± 9.9% and 21.3 ± 7.4 g, respectively, for octopuses fed squid, hake or crayfish, and were always higher for octopuses fed squid and hake compared to crayfish. The results indicate that crayfish is not an adequate replacement for the usual prey to fatten octopus, even considering its much lower market price.     

Effects of genetically modified and non‐genetically modified soybeans with different heat treatments on growth and health of Cyprinidae species with different feeding habits

An 8‐week growth trial was conducted to investigate the effects of non‐genetically modified (nGM) soybean (Youchun 06‐1) and genetically modified (GM) soybean (Roundup Ready^®) with and without a heat treatment on the growth and health of three Cyprinidae species with different feeding habits (grass carp Ctenopharyngodon idellus, gibel carp Carassius auratus gibelio, and black carp Mylopharyngodon piceus; body weight: 283.0 ± 2.0 g, 60.5 ± 0.7 g, and 261.4 ± 3.1 g). Five diets (FM, nGMS, hnGMS, GMS, and hGMS) were made with fishmeal and full‐fat soybean (Youchun 06‐1, heat‐treated Youchun 06‐1, Roundup Ready^®, and heat‐treated Roundup Ready^®). The FM diet contained fishmeal as the sole protein source. The experimental diets contained full‐fat soybean meal as 60% of dietary protein. Both temperature (60, 80, 100, and 120°C) and duration (1 and 2 hr) of heating influenced trypsin inhibitor activity (TIA) and protein solubility of nGM soybean and GM soybean. The TIA in GM soybean was higher than that in nGM soybean. After heating at 120°C for 2 hr, each amino acid of soybean treatment showed a slight decline. Neither source of soybean nor heat treatment affected the growth performance, feed utilization, chemical composition of the whole fish body and muscle, and lipase or amylase activity of the three Cyprinidae species (p > 0.05). Heat treatment of soybean meal slightly increased the plasma antioxidant capacity of the three fish and plasma cholecystokinin of black carp and grass carp. The unheated soybean treatment adversely affected the height and density of black carp intestinal villi, and all the soybean treatments caused disruption of the grass carp intestinal epithelium.     

Shrimp shell as a potential sorbent for removal of arsenic from aqueous solution

This study examined the ability of shrimp shell to remove arsenic (As) from aqueous solutions. The shells of two species of shrimp, black tiger shrimp Penaeus monodon and white shrimp Litopenaeus vannamei, were chosen to be the sorbents. Laboratory exposure experiments estimated uptake and depuration rate constants (i.e., k ₁ and k ₂) as well as the bioconcentration factors (BCF) of the shells of the two shrimps. A first-order one-compartment model was presented to describe the uptake kinetics of As in shrimp shell. The resulting k ₁, k ₂, and BCF values of black tiger shrimp were 0.034–1.722 ml/g/day, 0.007–0.345 g/g/day, and 5.08 ± 1.56 ml/g, while those for white shrimp were 0.053–0.523 ml/g/day, 0.011–0.237 g/g/day, and 3.95 ± 1.88 ml/g, respectively. The sorption capacities of black tiger shrimp shell and white shrimp shell were 1.08 × 10⁻⁴–6.66 × 10⁻³ and 1.04 × 10⁻⁴–3.26 × 10⁻³ mmol/g, respectively. The sorption capacity of shrimp shell increased with the initial As concentration in water. Shrimp shell, as a waste material, could be potentially used for the removal of As from an aqueous medium. Although the As-removal capacity of shrimp shell was lower than those of natural and chemical sorbents, using shrimp shells as sorbents is less expensive and could increase the additional value of shrimp products.     

Physiological characteristics and stress resistance of great sturgeon (<Emphasis Type="Italic">Huso huso</Emphasis>) juveniles fed with vitamins C,E, and HUFA-enriched <Emphasis Type="Italic">Artemia urmiana</Emphasis> nauplii

This study was carried out to examine the effect of Artemia urmiana nauplii enriched with HUFA, and vitamins C and E on stress tolerance, hematocrit, and biochemical parameters of great sturgeon, Huso huso juveniles. Cod liver oil (EPA 18% and DHA 12%), ascorbyl-6-palmitate and α-tocopherol acetate were used as lipid, and vitamin C and E sources, respectively. Beluga juveniles at the stage of first feeding (69.7 ± 5.9 mg body weight) were randomly divided into five treatments and three tanks were assigned to each diet. All fish groups were fed non-enriched Artemia for the initial 5 days and then fed enriched Artemia for 7 days. Juveniles were fed with Artemia enriched with HUFA + 20% vitamin C (C group); HUFA + 20% vitamin E-enriched Artemia nauplii (E group); HUFA + 20% vitamin C + 20% vitamin E (C and E group); HUFA without vitamins (HUFA) and non-enriched Artemia (control). After the period of enrichment, Juveniles were fed with Daphnia sp. from the 13th to the 40th day. At day 40, the fish were transferred directly from fresh water (0.5 ppt) to brackish water (6 ppt for 4 days and 12 ppt for 2 days) and warm water (from 27 to 33°C) to evaluate juvenile resistance to salinity and thermal shocks. Moreover, all treatments were separately exposed to freshwater in tanks with the same capacity as used for osmotic and thermal tests (as fresh water control). The addition of vitamins C, E, and C + E to HUFA significantly increased fish resistance to 12 ppt salinity and temperature stress tests, whereas survival was not significantly different among challenges at 6 ppt. There was no significant difference in the hematocrit index under stress conditions. Enrichment had significant influence on plasma Na⁺ level in the C group on the 4th day at 6 ppt. Na⁺ and Ca²⁺ concentrations in C, E, and C and E groups on the 1st day at 12 ppt, and Ca²⁺ level in E group on the 2nd day at 12 ppt were lower than the other groups. The glucose level in the C and C and E groups was lower than the other treatments on the 1st day at 12 ppt and the 2nd day at 33°C. Regardless of Artemia enrichment, plasma ions (Na⁺, K⁺, Ca²⁺, and Mg²⁺) and glucose concentrations in fish exposed to salinity stress tests were higher than fish in fresh water. Glucose concentration in plasma also increased after 2 days at 33°C. Although most of our results were not significantly different, the use of vitamins C, E, and HUFA in Artemia enrichment can improve Juveniles tolerance under stress conditions, and regardless of enrichment, these data show that beluga juveniles are partly sensitive to high salinity and temperature.     

In vitro oocyte maturation and blood‐circulating steroid hormones in Sterlet (Acipenser ruthenus) and Siberian sturgeon (Acipenser baeri)

The oocyte of Sterlet (Acipenser ruthenus) and Siberian sturgeon (Acipenser baeri) was evaluated after incubation with 17α, 20β‐Dihydroxyprogesterone (DHP) in artificial media. Concentration of testosterone (T), progesterone (P4) and 17‐β estradiol (E2) in blood and their relation to fertilization rate were measured during experimental period. The oocytes at the same developmental stages were incubated in SIS (based on sturgeon blood ionic composition), RM2 (Ringer solution modified for sturgeons) and L‐15 (Leibovitz medium) artificial mediums in the presence of 1 μg/ml of 17α, 20β‐Dihydroxyprogesterone for 12, 18 and 24 hr. The result of this study demonstrated that fertilization rate decreased with the increasing duration of incubation in all mediums. In the Siberian and Sterlet sturgeon, the highest fertilization rate was observed in the oocytes incubated in RM2 (0.81 ± 0.4) and SIS (0.44 ± 0.8) mediums for 12 hr respectively. In the Siberian sturgeon, fertilization rate decreased significantly (p < .05) with incubation time in all three mediums. In Starlet, fertilization rate decreased significantly (p < .05) with time and reached a minimum after 24 hr (0.16 ± 0.1) in L‐15 medium. No significant (p > .05) differences were observed between blood plasma hormones and fertilization rate. The results of this study indicated that 17α, 20β‐Dihydroxyprogesterone is essential for induction of in vitro oocyte maturation in these species.     

Evaluation of cGnRH IIa for induction spawning of two ornamental Epalzeorhynchos species

The administration of exogenous hormones for induction spawning of captive fish is a common practice throughout the finfish aquaculture industry. Induced spawning protocols for ornamental cyprinids commonly rely on sGnRH IIIa, commercially available as Ovaprim. In this study, the efficacy of an alternative spawning aid, cGnRH IIa, was evaluated relative to Ovaprim, in two commonly cultured ornamental Cyprinids: the Redtail Sharkminnow Epalzeorhynchos bicolor and the Rainbow Shark E. frenatum. Broodstock of each species were injected with either a positive control (0.5 µl/g Ovaprim), a negative control (propylene glycol) or one of three doses of cGnRH IIa (50, 100, or 200 µg/kg). Following spawning aid injection, ovulation success, fecundity, fertilization success, embryo diameter, hatch success and larval notochord length were evaluated. Ovulation success (82 ± 8%) was statistically similar to the positive control group for all experimental doses of cGnRH IIa in E. bicolor, while 50 and 100 µg/kg cGnRH IIa doses resulted in significantly higher ovulation success (100.0% and 83.0% respectively) than the positive control (17.0%) in E. frenatum. All other parameters did not vary significantly among treatments. These results indicate cGnRH IIa can be successfully used as a spawning aid in both species.     

Lack of arterial PO<Subscript>2</Subscript> downregulation in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.) during long-term normoxia and hyperoxia

Regulation of arterial partial pressure of O₂ (P_aO₂) in Atlantic salmon (Salmo salar) was investigated during resting conditions in normoxic and hyperoxic water. Dorsal aorta cannulated adult Atlantic salmon (1.2–1.6 kg, n = 8) were exposed to 2 week sequential periods of normoxia [16.7 ± 1.1 kPa (mean ± SD)] and hyperoxia (34.1 ± 4.9 kPa) in individual tanks containing seawater (33.7 ± 0.2 ppt) at stable temperature conditions (8.7 ± 0.7°C) and a light regime of L:D = 12:12. Tank design and sampling procedures were optimized to provide suitable shelter and current for the fish, and to allow repeated, undisturbed sampling of blood from free-swimming fish. Fish were sampled regularly through the experimental period. P_wO₂, P_aO₂, blood ion composition (Na⁺, K⁺, Cl⁻), acid–base status (pH, PCO₂, HCO₃ ⁻), haematocrit and glucose were measured. The most frequently observed P_aO₂ values were in the range of 60–80% of P_wO₂, both during normoxia and hyperoxia, and P_aO₂ values were significantly lower during normoxia than during hyperoxia. Blood pH, PCO₂ and HCO₃ ⁻ were significantly elevated during hyperoxia, while, Na⁺, Cl⁻ and Hct were significantly lower. K⁺ and glucose showed no significant differences. This study demonstrates a lack P_aO₂ regulation in Atlantic salmon to low partial pressures, in contrast to previous reports for many aquatic gill breathing animals. Both during normoxia and hyperoxia, P_aO₂ reflects P_wO₂, and alterations in external PO₂ consequently result in proportional arterial PO₂ changes. Physiological adaptation to hyperoxia, as illustrated by changes in several blood parameters, does not include down-regulation of P_aO₂ in Atlantic salmon. The lack of P_aO₂ regulation may make Atlantic salmon vulnerable to the oxidative stress caused by increased free radical formation in hyperoxic conditions.     

Composition of ovarian fluid in endangered Caspian brown trout, <Emphasis Type="Italic">Salmo trutta caspius</Emphasis>, and its effects on spermatozoa motility and fertilizing ability compared to freshwater and a saline medium

In this study, ovarian fluid composition and its effects on the motility and fertilizing ability of sperm were studied in endangered Caspian brown trout, Salmo trutta caspius, and were compared with a saline activation medium (125 mM NaCl, 30 mM Glycine, 20 mM Tris–HCl, pH = 9.0) and freshwater as the control. The ovarian fluid was composed of sodium 164.4 ± 4.4 mM l⁻¹, potassium 1.8 ± 0.1 mM l⁻¹, calcium 0.6 ± 0.1 mM l⁻¹, magnesium 0.4 ± 0.02 mM l⁻¹, chloride 127.4 ± 5.9 mM l⁻¹, total protein 389.5 ± 89.6 mg 100 ml⁻¹, cholesterol 9.3 ± 1.2 mg dl⁻¹, and glucose 3.3 ± 0.2 mM l⁻¹. The percentage of motile spermatozoa and the duration of sperm motility were significantly higher in ovarian fluid (62 ± 3%, 74.6 ± 0.8 s) than freshwater (35 ± 4%, 44 ± 1 s), but they did not differ significantly from saline medium (56 ± 3%, 74.3 ± 0.7 s) (P > 0.05). Higher eyeing rates were observed after the activation of sperm in ovarian fluid and saline solution than freshwater when 35,000 or 350,000 spermatozoa per egg were added into the activation media. However, no significant differences were observed at higher concentrations of spermatozoa per egg (730,000) (P > 0.05). Also, this study showed that the ovarian fluid composition can be considered as a species-specific character among salmonid fishes. As a conclusion, the results of this study recommend the use of ovarian fluid or the saline solution as an activation medium in the artificial reproduction of Caspian brown trout.     

Characterization of digestive enzymes protease and alpha‐amylase activities in the thick‐lipped grey mullet (Chelon labrosus,Risso 1827)

Activities of pepsin, trypsin, chymotrypsin, alpha‐amylase and lipase, as well as their optima and stability to pH and temperature, were determined in digestive extracts of thick‐lipped grey mullet, Chelon labrosus, of three different sizes: Group 1 (45.2 ± 3.0 g), Group 2 (180.9 ± 4.2 g), and Group 3 (328.5 ± 43.3 g). SDS‐PAGE zymograms were also used to assess the role of serine proteases in the digestive tract of C. labrosus. On the other hand, possible changes in the digestive enzyme profile of C. labrosus during development were observed, with a comparatively lower pepsin activity, higher activities of alkaline proteases and alpha‐amylase and no lipase activity recorded in pre‐adult specimens. It is suggested that these variations are linked to the changes in diet composition with age, moving from a partly carnivorous to a more herbivorous feeding habit.