Ovulation and spawning induction in Caspian kutum Rutilus frisii kutum by administration of GnRHa and catecholaminergic pharmaceutical compounds with Ovaprim

The acceleration of final oocyte maturation and ovulation is recognized as a principal process in Caspian kutum Rutilus frisii kutum, a commercial and valuable species in Caspian Sea. The aim of this study was to evaluate the effects of catecholaminergic pharmaceutics which include Salbutamol (β₂‐adrenergic receptor agonist), Metoprolol (β₁‐adrenergic receptor antagonist), Clozapine and Olanzapine (the third generation of D₄ and D₂ dopamine receptor antagonists) which can be more efficient than the first and second generations, in combination with Buserelin ([D‐Ser(tBu)⁶,Pro⁹‐NEt]‐GnRHa) and Ovaprim (D‐Ala⁶,Pro⁹‐Net)‐sGnRH+Domperidone on ovulation and spawning induction in Caspian kutum. One hundred eight adult females were injected once with Buserelin Acetate (BUS) 5 μg kg^?1 BW, Ovaprim 0.5 mL kg^?1 BW, Salbutamol Sulphate (SLB) 4 mg kg^?1 BW, Olanzapine (OLZ) 5 mg kg^?1 BW, Clozapine (CZ) 12 mg kg^?1 BW and Metoprolol Tartrate (MTP) 5 mg kg^?1 BW, being divided into 12 groups: group 1, intact (negative control); group 2, Ovaprim (positive control); group 3, OLZ+SLB; group 4, OLZ+MTP; group 5, OLZ+BUS; group 6, CZ+SLB; group 7, CZ+MTP; group 8, CZ+BUS; group 9, OLZ+BUS+SLB; group 10, OLZ+BUS+MTP; group 11, CZ+BUS+SLB and group 12, CZ+BUS+MTP (N = 9). The results showed that the highest mean value in ovulation success, ovulation index, fertilization success, relative fecundity and the number of eggs belonged to Ovaprim treatment. On the other hand, spawning was successful in OLZ+BUS+SLB and CZ+BUS treatments (P < 0.05), whereas it was lower in CZ+MTP, CZ+BUS+MTP and OLZ+BUS than the other treatments. Therefore, it could be concluded that Clozapine and Olanzapine potentiated the effect of Buserelin treatment in ovulation and spawning induction, while Metoprolol blocked the stimulatory effects of GnRHa, Clozapine and Olanzapine. Salbutamol also can amplify stimulatory effect of all mentioned pharmaceutics.     

Gonadal recrudescence and induced spawning in Barbus altianalis

Gonadal recrudescence in Barbus altianalis from River Nile, Jinja (33°05E; 0°45N) Uganda was studied prior to induced spawning. Oocytes in B. altianalis were found to develop in cohorts suggesting batch spawning throughout the year with pronounced spawning activities coinciding with rainfall peaks of April and September. There was a strong positive correlation between rainfall and gonadosomatic index (r_S = 0.75, P = 0.008). B. altianalis had low fecundity (6.0 ± 2.0 per gram) and large‐sized eggs (2.97 ± 0.1 mm). Induced spawning was successful with exogenous hormones and water current. Results from this study suggested that total striping may not be appropriate, but larger eggs would confer high survival of offsprings under optimal conditions. The success registered in induced spawning of B. altianalis provides a breakthrough in seed production for multiplication and culture of this economically important fish species.     

Characterization of spaC‐type Erysipelothrix sp. isolates causing systemic disease in ornamental fish

Since 2012, low‐to‐moderate mortality associated with an Erysipelothrix sp. bacterium has been reported in ornamental fish. Histological findings have included facial cellulitis, necrotizing dermatitis and myositis, and disseminated coelomitis with abundant intralesional Gram‐positive bacterial colonies. Sixteen Erysipelothrix sp. isolates identified phenotypically as E. rhusiopathiae were recovered from diseased cyprinid and characid fish. Similar clinical and histological changes were also observed in zebrafish, Danio rerio, challenged by intracoelomic injection. The Erysipelothrix sp. isolates from ornamental fish were compared phenotypically and genetically to E. rhusiopathiae and E. tonsillarum isolates recovered from aquatic and terrestrial animals from multiple facilities. Results demonstrated that isolates from diseased fish were largely clonal and divergent from E. rhusiopathiae and E. tonsillarum isolates from normal fish skin, marine mammals and terrestrial animals. All ornamental fish isolates were PCR positive for spaC, with marked genetic divergence (<92% similarity at gyrB, <60% similarity by rep‐PCR) between the ornamental fish isolates and other Erysipelothrix spp. isolates. This study supports previous work citing the genetic variability of Erysipelothrix spp. spa types and suggests isolates from diseased ornamental fish may represent a genetically distinct species.     

Pharmacokinetics and physio‐metabolic response of single and multiple dose of fenbendazole in Labeo rohita (Hamilton, 1822) fingerlings

An experiment was carried to determine the plasma fenbendazole (FBZ) concentration and physio‐metabolic responses in juveniles of Labeo rohita (90 ± 4 g) after oral administration of single doses at 10, 20 and 50 mg, 20 mg FBZ/kg b.wt. in multiple times on 1st, 3rd and 7th day. The blood samples were collected at 0.5, 1, 2, 4, 8, 12, 24, 30, 48, 72, 96 and 120 hr, after single‐dose administration, and regularly (upto 15 day) in multiple dose. Plasma FBZ concentration was determined up to the limit of detection (LoD) of 0.09 µg/ml by HPLC. There was no parent drug detected in plasma for administration of 10 mg FBZ/kg b.wt. The drug attained the peak concentration (C_max) 1.85 and 3.09 µg/ml in plasma at 4 hr (T_max) after administration of 20 and 50mg FBZ/kg b.wt. respectively. Plasma FBZ was detectable up to 96 and 120 hr with concentration 0.09 ± 0.007 and 0.098 ± 0.006 µg/ml, respectively, after single‐dose administration of 20 and 50mg/kg b.wt. In case of multiple‐dose administration, the maximum concentration of FBZ was 1.01 ± 0.03 µg/ml on 7th day that was less than to the single dose at 50 mg/kg b.wt. However, FBZ was detected up to 11 day after multiple doses. The study revealed that the hepatic antioxidant enzymes activities like superoxide dismutase, catalase and glutathione‐S‐transferase were significantly affected by increasing FBZ in single and multiple doses. The results of the present study could reveal that single‐ or multiple‐oral administration of FBZ at 20 mg/kg b.wt. in feed as antihelminthic drug in L. rohita could be considered as the safe dose.     

Year‐round natural spawning,early development,and the effects of temperature,salinity and prey density on captive ornate goby Istigobius ornatus (Rüppell, 1830) larval survival

Natural spawning, early development and larviculture of the ornate goby Istigobius ornatus in captivity were studied for the first time. I. ornatus spawned 46 times from 31 October 2013 to 31 October 2014. Fecundity ranged from 246 to 10,214 eggs per clutch, with an average hatching rate of 77.8% ± 9.9% (M ± SEM). Fertilized eggs (1.31–1.54 × 0.46–0.50 mm in diameter) were adhesive demersal and oval‐shaped. Embryonic development lasted 84 hr at 27.5 ± 0.5°C. Newly hatched larvae [2.12 ± 0.04 mm in total length (TL)] transformed to the juvenile stage completely when TL was 7.79 mm. Effects of different water temperatures (24, 28 and 32°C) and salinities (10, 15, 20, 25, 30, 35 and 40 g/L) on per cent survivals (%) and survival activity indices (SAIs) were tested. Survival was not significantly different under different temperatures; SAIs was significantly higher at 28°C. Larvae showed the significantly higher survival and SAIs at salinities 10–30 g/L than at 35 and 40 g/L. Effect of different prey densities on survival was significantly higher in 7 days post hatch larvae fed 20 and 30 rotifers/ml. These findings could guide future programs in captive breeding technology development and commercial production of other marine ornamental gobies.     

Comparison of mudcrab‐based brackishwater polyculture systems with different finfish species combinations in Sundarban,India

To compare production and economic performance of polyculture systems with different species combinations, a 210‐day trial was carried out. In the first combination (T1), milkfish (Chanos chanos) and mudcrabs (Scylla serrata), and in the second (T2), mullets (Mugil cephalus, Liza tade and Liza parsia at 0.5:0.5:0.5 ratio) and mudcrabs were stocked keeping fish and mudcrabs at 15,000 numbers per ha, respectively, in both treatments. The finfish were fed floating pellet at 2%–3% and mudcrabs were fed fresh and farm made feed at 5%–8% body weight. Growth parameters of mudcrabs were similar in both T1 (407.64 ± 105.78 g) and T2 (418.89 ± 105.24 g), with no significant differences. Among finfish, M. cephalus attained highest final body weight, 241.55 ± 26.44 g followed by milkfish, 200.46 ± 11.82 g whereas lowest growth noticed in L. parsia (63.69 ± 6.62 g). Length–weight analysis of fish indicated negatively allometric growth (b < 3) for grey mullets, parsia and milkfish while L. tade recorded perfect cube low (b = 2.99). Male mudcrabs recorded positive allometric (b = 3.3) and female crab exhibited negative allometric growth (b = 2.68). The total productivity was 4,533 and 3,694 kg/ha with mudcrab contributes 53.69% and 60.56% to the total productivity in T1 and T2 respectively. The economic analysis indicated benefit–cost ratio (BCR) of 1.57 and 1.73 in T1 and T2 respectively with 10% and 35% insignificant increase (p > 0.05) in BCR and profit per kg, respectively, in T2 compared with T1. The study elucidates polyculture of mudcrabs with finfish can be a taken up as a profitable venture for sustainable diversification of brackishwater farming in Sundarbans.     

Biofloc technology in intensive broodstock farming of the pink shrimp Farfantepenaeus duorarum: spawning performance,biochemical composition and fatty acid profile of eggs

A 45‐day trial was performed to evaluate the effect of biofloc technology (BFT) with or without fresh food (FF) supplementation during pre‐maturation period on Farfantepenaeus duorarum spawning performance, biochemical composition and fatty acid profile of eggs as compared with conventional clear‐water system (CW+FF). Females raised in biofloc and that received FF supplementation (FLOC+FF) achieved better spawning performance in terms of number of eggs per spawn (49 × 10³), number of eggs per spawn per g of spawner's body weight (2.1 × 10³) and egg size (~275 μm) as compared with CW+FF (23 × 10³, 1.1 × 10³ and 263 μm respectively), but both treatments did not vary from FLOC (P > 0.05). High spawning activity was also observed in biofloc system as compared with clear‐water system as shown in number of spawns per ablated female (2.2–3.0 versus 0.6) and percentage of females that spawn at least once (80–82 versus 25%). Biochemical composition of eggs presented no significant differences among treatments. FA profile of eggs indicated that high spawning activity performed by females in FLOC+FF treatment was reflected in lower mean levels of EPA, DHA and sum of polyunsaturated fatty acids (n‐3) and (n‐6). The better reproductive performance demonstrated by females raised in biofloc justified the application of this technology in F. duorarum broodstock.     

Conditioning,maturation and year‐round natural spawning of orange‐spotted grouper,Epinephelus coioides (Hamilton, 1822), in recirculating aquaculture system

The present experiment was aimed at studying the conditioning, maturation and natural spawning of orange‐spotted grouper, Epinephelus coioides, in a recirculatory aquaculture system (RAS). Thirty fish (n = 30; 3.35 ± 0.05 kg) were stocked in a circular tank of 125 m³ capacity fitted with an RAS for conditioning and broodstock development. After 15 days, 15 fish were implanted with 17 α methyl testosterone and letrozole at the rate of 5 mg and 0.2 mg/kg body weight, respectively, for conversion from female to male. The gonadal development started after 1 month, and by 90th day, 63.53 ± 3.78% and 2.07 ± 0.84% of the oocytes attained a size of 400–500 μm and 500–600 μm respectively. Natural spawning commenced in the RAS from 4th month onward after stocking and spawning continued round the year. The spawning pair showed courtship behaviour with a typical vertical burst of swimming just before release of gametes. The total number of eggs spawned during 1 year was 47.23 million with spawning frequency varying form 5 to 13 times per month. The association of spawning events with new moon day (lunar cycle) weakened as time progressed. The mean monthly fertilization and hatching rates varied from 77.80 ± 3.34% to 83.70 ± 1.76% and 82.80 ± 4.21% to 88.33 ± 1.39% respectively. The study proved that RAS is an efficient system that provides a stable, controllable and conducive environment for year‐round natural breeding of orange‐spotted grouper.     

An investigation on the gamete quality of Black Sea trout (Salmo trutta labrax) broodstock fed with mealworm (Tenebrio molitor)

In this study, the effect of combined feeding Black Sea trout (Salmo trutta labrax) broodstock with commercial feed and mealworm larvae (Tenebrio molitor) for 50 days before the breeding period in order to evaluate the gamete quality. While the control group was fed with commercial feed, mealworm larvae were given as an additional protein source 2 days a week and 3 days a week to the experimental groups formed from female and male individuals. In addition to the growth parameters, the number of eggs (number/individual) and the egg diameter (mm), sperm volume (ml), density of spermatozoa (×10⁹ cell/ml), total motility (%), progressive motility (%) and average curvilinear velocity (VCL, µm/sec) values were determined. At the end of the study, the highest live weight gain was found as 46.2 g in the control group of male. The egg diameters were 4.3 ± 1.8 mm and 4.5 ± 1.4 mm in the worm treatment groups fed twice and three times with mealworm respectively. As a result, it was determined that feeding fish with mealworm larvae as an addition to the commercial diet in female individuals did not affect the amount of eggs (p > .05), but the egg diameters were significantly smaller in the control group than the mealworm groups (p < .05). In spermatological characteristics, only the amount of sperm was different between the groups (p < .05); all other parameters were found to be similar to each other with no statistical differences (p > .05).     

Dietary Houttuynia cordata leaf extract and meal enhances the immunity and expression of immune genes in Labeo rohita (Hamilton, 1822)

A feeding trial was conducted to explore the effect of dietary Houttuynia cordata leaf extract (HCLE) and leaf meal (HCLM) on immunological responses and expression of interferon‐gamma (IFN‐γ) and tumour necrosis factor‐alpha (TNF‐α) gene in Labeo rohita fingerlings. Six isonitrogenous (350 g/kg CP) and isocaloric (17 MJ/kg DE) purified experimental diets were formulated with Houttuynia cordata leaf extract and leaf meal comprising control, C (0 g/kg HCLE and HCLM), E2.5 (2.5 g/kg HCLE), E5 (5 g/kg HCLE), E10 (10 g/kg HCLE), M10 (10 g/kg HCLM) and M20 (20 g/kg HCLM). Labeo rohita fingerlings (3.37 ± 0.23 g) were distributed in six experimental groups in triplicates following the complete random distribution. Fish were fed twice daily with respective experimental diets for a period of 60 days. A significantly (p < .05) lower lactate dehydrogenase, malate dehydrogenase, superoxide dismutase and catalase activities were registered in supplemented groups compared with control group, while respiratory burst and lysozyme activities were significantly (p < .05) higher in E10 group compared with other experimental groups. Haemoglobin, total leucocyte count, total erythrocyte count and haematocrit values were significantly (p < .05) higher in E10 group. The expression of IFN‐γ and TNF‐α in both the kidney and liver was significantly up‐regulated in leaf extract and meal supplemented groups with the highest expression in the fish of E10 group. Overall, these results suggest that the dietary supplementation of ethanolic extract of the Houttuynia cordata leaf at 10 g/kg level can enhance the immune response of L. rohita fingerlings.     

Effects of dietary Nutrafito Plus on growth,haemotological parameters and total ammonia‐nitrogen excretion of juvenile striped catfish Pangasianodon hypophthalmus

A feeding trial was performed to determine the effect of a commercial source of Yucca schidigera and Quillaja saponaria extracts (Nutrafito Plus^®), in practical diets, on striped catfish Pangasianodon hypophthalmus growth, feed utilization, body composition, total ammonia‐nitrogen (TAN) excretion and haematological parameters. Four experimental diets were supplemented with Nutrafito Plus^® at 0%, 0.01%, 0.02% and 0.03% (diets: control, N01, N02 and N03 respectively). Three replicate groups of striped catfish, with initial mean weight of 1.78 ± 0.05 g, were fed one of the four diets for 12 weeks. The specific growth rate (SGR) and final weight of fish fed diet N03 was significantly higher (P < 0.05) than fish fed the other dietary treatments. The growth performance of fish fed diets N01 and N02 were not significantly different compared to fish fed the control diet. Striped catfish fed diet N03 had improved feed conversion ratio (FCR), net protein utilization (NPU) and protein efficiency ratio (PER) than the control (P < 0.05). The inclusion of the high dietary level of Yucca schidigera and Quillaja saponaria, diet N03, reduced TAN compared to all groups. Dietary inclusion of Yucca schidigera and Quillaja saponaria at all levels investigated did not affect the whole body proximate composition of the striped catfish (P > 0.05). The packed cell volume (PCV) and haemoglobin level in fish fed diet N03 was significantly higher than in the fish fed the control diet. The present study demonstrates that dietary inclusion of Yucca schidigera and Quillaja saponaria induced positive effects on growth performance and haematological parameters and decreased TAN excretion in striped catfish.     

Determination of grapefruit (Citrus paradisi) peel extract bio‐active substances and its application in Caspian white fish (Rutilus frisii kutum) diet: Growth,haemato‐biochemical parameters and intestinal morphology

In this study, total phenolic and flavonoid contents of grapefruit peel extract (GPE) were equal to 117.3 ± 0.3 µg of gallic acid/mg and 39.30 ± 0.1 µg of quercetin/mg respectively. Caspian white fish (n = 180, 4 ± 0.9 g body weight) was fed with supplemented diets, including 0, 6.25, 12.5 and 25 mg of GPE/kg for 60 days at 25 ± 1°C. The growth performance was markedly improved in fish fed with 25 mg/kg of GPE compared to others (p < 0.05). Moreover, fish fed with 25 mg/kg of GPE showed a significant increase in red blood cell (2.65·10⁶ cell/mm³), white blood cell (17.75·10³ cell/mm³), packed cell volume (48%) and haemoglobin concentrations (8.75 g/dl) compared to the control (p < 0.05). However, the highest alanine aminotransferase (140 U/L), alanine transaminase (14.5 U/L), and alkaline phosphatase (18.5 U/L) were observed in control group. Morphological analysis of intestine revealed the highest amount of villus width (8.4 µM), height (32.86 µM) and surface area (342.7 µM²) in fish fed with 25 mg/kg of GPE (p < 0.05). In conclusion, supplementing feed with GPE at 25 g/kg can improve growth performance and haemato‐biochemical parameters of Caspian white fish fry.     

Effect of Ocimum sanctum Linn. (Tulsi) extract on the immunity and survival of Labeo rohita (Hamilton) infected with Aeromonas hydrophila

A 60 days study was conducted to evaluate the efficacy of water extract of Ocimum sanctum Linn. leaf on the immune response and disease resistance of Labeo rohita fingerlings against the Aeromonas hydrophila infection. Ocimum sanctum extract was incorporated in the diets (at 0.0%, 0.05%, 0.1%, 0.2%, 0.5% and 1%) of Labeo rohita, rohu fingerlings (6.6 ± 0.013 g). After 42 days blood, plasma and serum were sampled to determine super oxide anion production, lysozyme activity, total immunoglobulin in plasma, blood glucose, serum total protein, albumin, globulin, albumin:globulin ratio, WBC, RBC, haemoglobin content. Fish were challenged with A. hydrophila after 42 days and mortalities were recorded over 18 days post infection. The results demonstrate enhanced super oxide anion production, lysozyme activity, total immunoglobulin in plasma, serum total protein, globulin, total RBC counts, total WBC counts and haemoglobin content (P < 0.05) in treatments group compared with control group. Dietary O. sanctum extracts of 0.2% showed significantly (P < 0.05) higher protection relative percentage survival (RPS 40.00 ± 5.773%) against A. hydrophila infection than control. These results indicate that O. sanctum leaf extract stimulates the immunity and makes L. rohita more resistant to bacterial infection (A. hydrophila).     

Induction of spawning in Pacific white shrimp Litopenaeus vannamei (Boone, 1931) by injection of its molt inhibiting hormone isoform II produced in E. coli

The optimization of reproductive parameters in shrimp farming continues to be a challenge for most producing countries. Although the crustacean neuropeptides have been studied extensively in the last two decades, the functions of most of these neuropeptides remained putative. Among them, molt‐inhibiting hormone isoform II (MIH II) has shown an important role in vitellogenesis. In this study, the cDNA encoding mature MIH II peptide was isolated by RT‐PCR from the L. vannamei eyestalk. The cDNA was cloned into pET28a bacterial expression vector. Recombinant MIH II was obtained in the form of insoluble inclusion bodies and purified to ~88% purity. Two doses of rMIH II and a negative control group were assayed in vivo. The stages of ovarian maturation and spawning were recorded during 72 hr post‐injection. The results showed that ovarian maturation occurred approximately in 9% and 33% of females injected with rMIH II at the doses of 300 and 600 ng/gbw respectively. Neither maturation nor spawning was detected in the negative control group. Females injected with 600 ng/gbw, which showed vitellogenic stages III and IV, spawned. These preliminary results argue that the hormone rMIH II could be a promising candidate to induce spawning in L. vannamei shrimp.     

A new potential therapeutic remedy against Aeromonas hydrophila infection in rainbow trout (Oncorhynchus mykiss) using tetra,Cotinus coggygria

Different antibiotic‐based drugs are being used for the treatment of Aeromonas hydrophila infection in rainbow trout, and several studies emphasize the use of medicinal plants as immunostimulants for prophylactic measure against Aeromoniasis disease. However, therapeutic effects of aqueous methanolic extracts of tetra (Cotinus coggygria) against A. hydrophila in rainbow trout were not investigated. Four different concentrations of tetra extract (0 [control], 4, 8 and 12 mg/100 µl) and also two different positive control groups (florfenicol and doxycycline antibiotics) were administered orally using feeding needles to individual rainbow trout, Oncorhynchus mykiss of all experimental groups twice a day after intramuscular inoculation of A. hydrophila. The study period was for 10 days. On 0th, 3rd, 7th and 10th day, blood and tissues were collected from the fish and changes in humoral immune responses, haematology and immune‐related gene expressions were determined. In the study, superoxide radical production was decreased generally in all experimental groups except in 12 mg tetra and florfenicol treatments compared to control (p < .05). Lysozyme activity was generally decreased (p < .05), or no differences were observed in all experimental groups compared to the control. Myeloperoxidase activity was significantly increased in florfenicol‐treated fish group on 7th day (p < .05). Generally, myeloperoxidase activity showed an increase in almost all tetra‐treated groups. Haematological parameters increased but were not significantly high enough in treatments. Almost all immune‐related gene expressions were significantly enhanced on 3rd and 10th day of the study. Survival rate of 53.33% was found in control group. There were no significant differences in survival between control and 4 mg tetra‐treated group (p > .05). All the other groups' survival rate was significantly increased compared to control. The highest survival rate was found in florfenicol group (80%). In 12 mg tetra‐, doxycycline‐ and 8 mg tetra‐treated groups, survival rate was recorded as 74.44%, 70% and 70%, respectively. Our results suggest that tetra methanolic extract is an effective therapeutic remedy against A. hydrophila infection in rainbow trout at the dose of 24 mg/32.34 g body weight/day.     

Effects of dietary Bacillus pumilus on growth performance,innate immunity and digestive enzymes of giant freshwater prawns (Macrobrachium rosenbergii)

The present study was conducted to investigate the effects of Bacillus pumilus on the growth performance, innate immunity and digestive enzymes of the giant freshwater prawn, Macrobrachium rosenbergii. Prawns (mean weight: 1.81 ± 0.01 g) were fed different levels of B. pumilus at doses of 0, 1 × 10⁷, 1 × 10⁸ and 1 × 10⁹ CFU/g to produce one control group and three experimental groups (G1, G2 and G3), respectively. After a 60‐day experimental feeding period, the results showed that the final weight, weight gain rate (WGR) and specific growth rate (SGR) were significantly higher in experimental groups than in the control (p < 0.05), whereas these improvements did not differ significantly (p > 0.05) among experimental groups. The improvements in phagocytic activity, respiratory burst (RBs) activity and the activity of catalase (CAT), nitric oxide synthase (NOS) and acid phosphatase (ACP) were significantly higher in G2 and G3 than in the control and G1 (p < 0.05). Further, the activity of CAT, NOS and ACP significantly increased with an increase in the dietary level of B. pumilus (p < 0.05). The activity of phenoloxidase (PO) in G3 was significantly higher than that in the control (p < 0.05). The activity of amylase was significantly higher in the groups G2 and G3 than that in the control group. G3 improved the activity of protease compared with the control (p < 0.05). In conclusion, the results of this study indicated that the potential use of B. pumilusat a dose of 1 × 10⁸ CFU/g feed could improve the growth, immunity and digestive enzymes of M. rosenbergii.     

Improvement of reproductive indices,lysozyme activity,and disease resistance in live‐bearing ornamental fish,Poecilia latipinna using Artemia supplementation with treated yeast cell,Saccharomyces cerevisiae

In the present study, the effects of Artemia supplemented with 2‐β‐mercapto‐ethanol (β‐ME) treated yeast cell, Saccharomyces cerevisiae, on growth and reproductive performance, lysozyme activity, and disease resistance to Aeromonas hydrophila of freshwater ornamental species, Poecilia latipinna, were investigated. Within 60 days, molly fish were fed with three treatments including commercial food (T1), un‐supplemented Artemia (T2), and Artemia supplemented with β‐ME‐treated yeast cell (at concentration of 4 × 10⁷ CFU/L of water) (T3). After the feeding period, the fish were exposed to 100 μl of a suspension (1.1 × 10⁷ cells/ml) of A. hydrophila (BCCM5/LMG3279) and the cumulative mortality rates were recorded for 12 days. No significant difference was found between survival rate and growth performance of P. latipinna except for weight gain that was higher in fish fed through Artemia supplemented with β‐ME‐treated yeast cell compared to control group. Fecundity rate was significantly improved in fish fed using T3 with the maximum amount of 49.5 ± 2.29 per female (p < 0.05). Besides, lysozyme activity was significantly increased in group 3 (p < 0.05). Moreover, lowest fish mortality was significantly observed in this treatment (p < 0.05). In addition, the number of colonies formed by yeast cell in T3 (634 × 10³ CFU/g intestine) showed significant difference with other treatments (p < 0.05). In sum, Artemia enriched with β‐ME‐treated yeast improved reproductive indices, immune responses, and resistance against A. hydrophila of P. latipinna.     

Reproductive cycle of the rock oyster,Striostrea prismatica (Gray, 1825) from two locations on the southern coast of Ecuador

The reproductive cycle of the rock oyster Striostrea prismatica was determined at two fishing areas, General Villamil and Ayangue (located on the southern coast of Ecuador), between May 2012 and April 2013. Monthly sampling campaigns were performed at both locations. The tissues were histologically examined to determine gonadal index (GI), oocyte development, follicular area coverage and sex ratio. Surface seawater temperature, salinity and chlorophyll a concentration were measured during samplings. Our results show a similar annual reproductive pattern at both locations. The GI reached maximum values during the summer. Oysters reached highest ripeness in January and February, while spawning occurred in February–March. Gametogenesis was linked to a consistently increasing follicular area (from 1.3 ± 0.7% to 85.8 ± 7.8%) and associated to surface seawater temperature. Spawning coincided with warm water temperature fluctuations and a seawater salinity decrease. No correlation was found with Chlorophyll a concentration. The sex ratio of sampled populations was 1:1, suggesting that oysters sizing more than 10 cm in shell length present a stable sex proportion in the population. The diameter of mature oocytes was significantly reduced (32.7%) during histologically preparations in comparison to fresh oocytes. Our study provides useful information of environmental factors that may control the observed gametogenesis and spawning activity of S. prismatica.     

Growth and immune response of giant freshwater prawn Macrobrachium rosenbergii (De Man) postlarvae fed diets containing Chlorella vulgaris (Beijerinck)

A 50‐day growth trial was conducted to evaluate the efficacy of Chlorella vulgaris (Beijerinck) as an ingredient in the diets of giant freshwater prawn Macrobrachium rosenbergii (De Man) postlarvae (PL30). Immune response (total haemocyte count and prophenoloxidase activity) was also assessed by subjecting postlarvae to a challenge test against Aeromonas hydrophila (Chester) for 14 days. Iso‐nitrogenous and iso‐lipidic test diets were prepared using a fishmeal‐based‐positive control diet (D0) and four basal diets with inclusion levels of 2% (D2), 4% (D4), 6% (D6) and 8% (D8) C. vulgaris. Postlarvae of M. rosenbergii were randomly stocked (mean initial body weight of 0.19 ± 0.02 g) in 30‐L tanks in three replicates per dietary treatment for evaluation of growth performance. Another set of postlarvae (mean initial body weight of 1.25 ± 0.02 g) was randomly distributed in 95‐L tanks in three replicates per dietary treatment for the assessment of immune response. Results showed that specific growth rate was significantly higher (P < 0.05) in postlarvae fed D4 and D6. Variations in values for carcass protein, lipid, moisture and ash were also evident. Postlarvae fed diets with Chlorella showed increased prophenol oxidase activity and total haemocyte counts. Moreover, survival rate after challenge with A. hydrophila was significantly increased (P < 0.05). Inclusion of C. vulgaris in diets enhanced immune response and resistance of M. rosenbergii postlarvae against A. hydrophila infection.     

Increased gonad growth of the purple sea urchin (Strongylocentrotus purpuratus) fed the giant kelp (Macrocystis pyrifera) and the sea lettuce (Ulva lactuca) enriched with nutrients

Adult (Strongylocentrotus purpuratus) purple sea urchins were fed giant kelp (Macrocystis pyrifera) and sea lettuce (Ulva lactuca) macroalgae that were unenriched and enriched with nutrients. Urchins were fed over 10 weeks (5% body weight per day) under laboratory conditions to determine the effect of macroalgae species and treatment (enriched and unenriched) on growth, gonad index and gonad quality. Significant differences were observed when urchins were fed enriched Macrocystis pyrifera (MPE). Gonad weight increased from 0.7 ± 0.1 g (mean ± SD) to 5.9 ± 1 g, and gonad index increase from 3.5 ± 1.5% to 14.9 ± 2.3% over 10 weeks. Significant differences were also noted in gonad quality, and gonad colour and firmness were better in urchins that were fed unenriched M. pyrifera (MPUE) and MPE – whereas texture did not differ between M. pyrifera treatments and unenriched U. lactuca (ULUE). With regard to taste, gonads were sweeter in MPE‐fed individuals. Enriched U. lactuca (ULE) failed to grow gonads significantly; as a result, their quality was unacceptable for the market. These results suggest that S. purpuratus experience increased gonad production and quality in short periods when fed nutrient‐enriched M. pyrifera.