Effects of dietary calcium and phosphorus supplementation on the growth performance of juvenile spotted babylon <Emphasis Type="Italic">Babylonia areolata</Emphasis> culture in a recirculating culture system

A feeding experiment was conducted to determine the effects of dietary calcium and phosphorus, and the interaction between calcium and phosphorus, on the growth of juvenile spotted babylon, Babylonia areolata, cultured in a recirculating culture system. Nine isonitrogenous experimental diets supplemented with three levels of calcium (1, 4, and 7%) for each of three levels of phosphorus (1, 3, and 5%) were prepared using fish meal, squid meal, and shrimp meal as the main protein sources. Juveniles with an initial average body weight of 0.59 ± 0.09 g were fed to satiation once daily with one of the nine diets for 180 days. Absolute and specific growth rates were calculated for both shell length and whole wet body weight. Results showed that dietary calcium and phosphorus supplementation significantly affected the growth of juvenile spotted babylon (P < 0.05), but not survival and feed-conversion ratio. The specific growth rate in shell length (SGRL) ranged from 0.32 to 0.39% day⁻¹. No significant difference among phosphorus levels and no significant interaction between calcium and phosphorus in SGRL of the spotted babylon (P > 0.05) was found, but significant differences were observed among calcium levels, irrespective of phosphorus levels (P < 0.05). For 1 and 7% supplemental calcium, the spotted babylon had significantly higher SGRL than those fed diets supplemented with 4% calcium. However, the specific growth rate in body weight (SGRW) ranged from 0.91 to 1.19% day⁻¹ with no significant difference among calcium and phosphorus levels and no significant interaction between calcium and phosphorus (P > 0.05). Survival and feed-conversion ratio were not significantly affected by dietary calcium and phosphorus levels with ranges from 91.00 to 95.00% and 2.43 to 2.76, respectively. At the end of the experiment, shell abnormality of B. areolata was found for all feeding trials.     

Effect of dietary supplementation of brewers yeast and nucleotide singularly on growth,survival and vibriosis resistance on juveniles of the gastropod spotted babylon (<Emphasis Type="Italic">Babylonia areolata</Emphasis>)

This study was undertaken to evaluate the use of brewers yeast and nucleotides as a growth promoter and to provide vibriosis resistance for the juveniles of gastropod spotted babylon (Babylonia areolata). Juvenile spotted babylon (0.3 g initial weight) were randomly distributed at a density of 50 snails in 45-L aquaria and fed a basic diet (40% crude protein) containing two incremental levels of 1 and 2% brewers yeast and nucleotides singularly for 4 months. After the feeding trial, snails from each treatment were challenged by pathogenic bacteria Vibrio alginolyticus given by intramuscular injection and kept under observation for 5 days to record clinical signs and daily mortality rates. Results indicated that the snails fed with diets supplemented with brewers yeast or nucleotides exhibited significantly greater growth than those fed the basic diet (P < 0.05) and significantly better food conversion ratios compared to snails fed the basic diet (P < 0.05). These results indicated that dietary supplementation of brewers yeast or nucleotides, at least at the tested dosages, enhanced spotted babylon growth. Supplementing the diet with 1% brewers yeast promises to provide appropriate resistance to V. alginolyticus.     

Effects of dietary protein and lipid levels and protein to energy ratios on growth performance and feed utilization of hatchery-reared juvenile spotted babylon (<Emphasis Type="Italic">Babylonia areolata</Emphasis>)

A 120-day feeding trial was designed to determine the effects of different dietary protein and lipid levels and protein to energy ratio (P:E) on growth performance and feed utilization of hatchery-reared juvenile spotted babylon, Babylonia areolata, cultured under a flow-through seawater system. Six diets were formulated to contain three protein levels (18, 28, and 36%) and two lipid levels (10 and 15%) in a 3 × 2 factorial design with three replicates to provide six different dietary P:E ratios (50.17, 49.09, 68.50, 65.85, 88.66, and 85.36 mg protein/kcal). Each replicate was stocked with 50 snails (0.12 + 0.01 g, initial weight) and fed to satiation once daily. The results showed that survival was above 96% at the end of the feeding trial in all groups and was not affected by either dietary protein level or dietary lipid level. The highest significant (P < 0.05) growth and feed utilization were observed for juveniles fed diet with a P:E ratio of 88.66 kcal g⁻¹ diet. With respect to dietary protein and lipid levels, the highest (P < 0.05) values for growth and feed efficiency were observed for snails fed a diet containing 36% protein level and the same trend was observed for snails fed a diet with 10% lipid level. This results indicated that the diet containing 36% protein and 10% lipid level with a dietary P:E ratio of 88.66 mg protein/kcal would be suitable for optimum growth and feed utilization of B. areolata juveniles.     

Differences in fatty acid composition of egg capsules from broodstock spotted babylon, <Emphasis Type="Italic">Babylonia areolata</Emphasis>, fed a local trash fish and formulated diet under hatchery conditions

This study is the first attempt to condition broodstock Babylonia areolata using formulated diets under hatchery conditions. Samples of spotted babylon egg capsules from broodstock fed either a formulated diet or a local trash fish, carangid fish (Seleroides leptolepis) for 120 days were analyzed for proximate composition and fatty acid composition. The formulated diet contained significantly higher levels of arachidonic acid (20:4n − 6; ARA), eicosapentaenoic acid (20:5n − 3; EPA) and docosahexaenoic acid (22:6n − 3; DHA) than those of the local trash fish. The formulated diet also had significantly higher ratios of DHA/EPA and (n − 3)/(n − 6) PUFA than those of local trash fish but not for the ARA/EPA ratio. The compositions of egg capsules produced from broodstock fed formulated diet contained significantly more ARA, EPA and DHA compared to broodstock fed the local trash fish. The ARA/EPA and DHA/EPA ratios in egg capsules were significantly higher in the trash fish—fed group compared to those fed the formulated diet. However, (n − 3)/(n − 6) PUFA ratios in egg capsules produced from broodstock fed the formulated diet did not differ significantly compared to those from broodstock fed the local trash fish. The relatively low DHA/EPA, ARA/EPA and (n − 3)/(n − 6) ratios in the egg capsules produced from the formulated diet—fed broodstock B. areolata suggested that this diet is inferior, when compared to the traditional food of trash fish.     

Growth,food efficiency,and biochemical composition of juvenile spotted babylon <Emphasis Type="Italic">Babylonia areolata</Emphasis> (Link) fed on conventional trash fish and a formulated moist diet

A 120-day feeding trial was conducted to determine growth, feeding efficiency, and biochemical composition of juvenile spotted babylon Babylonia areolata (initial mean weight of 1.48 ± 0.06 g) fed on conventional trash fish and a formulated moist diet in a flow-through grow-out system. The snails readily accepted the formulated moist diet and showed good growth and survival during the trial. There were no significant differences in specific growth rate, feeding efficiency, and survival between the snails fed on trash fish or the formulated moist diet. At the end of the experiment, biochemical analysis of the snail flesh showed significant differences in flesh quality, with the snails fed on trash fish having a significantly higher crude protein compared to those fed on the formulated diet, but there was little difference in crude lipid content. Whole body tissue of snails fed on trash fish had significantly higher total amino acid, essential amino acid, and non-essential amino acid levels compared to those fed the formulated diet. This study showed that the formulated moist diet can be readily used for B. areolata growout for maximizing growth, survival, and feeding efficiency; however, further research is needed to improve the flesh quality particularly amino acid and fatty acid levels before its suitability for B. areolata is assured.     

Spatial learning-induced <Emphasis Type="Italic">egr</Emphasis>-<Emphasis Type="Italic">1</Emphasis> expression in telencephalon of gold fish <Emphasis Type="Italic">Carassius auratus</Emphasis>

The immediate-early gene (egr-1) expression was used to examine the neuron’s response in telencephalon of goldfish during spatial learning in small space. Fishes were pre-exposed in the experimental apparatus and trained to pick food from the tray in a rectangular-shaped arena. The apparatus was divided into identical compartments comprising three gates to provide different spatial tasks. After the fish learned to pass through the gate one, two more gates were introduced one by one. Fish made more number of attempts and took longer time (P < 0.05) to pass through the first gate than the gate two or three. This active learning induces the expression of egr-1 in telencephalon as established by western blot analysis. Subsequently, the fish learn quickly to cross the similar type of second and third gate and make fewer errors with a corresponding decline in the level of egr-1 expression. As the fish learned to pass through all the three gates, third gate was replaced by modified gate three. Interestingly, the level of egr-1 expression increased again, when the fish exhibit a high exploratory behavior to cross the modified gate three. The present study shows that egr-1 expression is induced in the telencephalon of goldfish while intensively acquiring geometric spatial information to pass through the gates.     

Growout of hatchery-reared juvenile spotted babylon (<Emphasis Type="Italic">Babylonia areolate</Emphasis> link 1807) to marketable size at four stocking densities in flow-through and recirculating seawater systems

Hatchery reared juvenile spotted babylon, Babylonia areolata, were raised in canvas rearing tanks at four stocking densities (100, 200, 300 and 400 ind. m^–2) in flow-through and recirculating seawater systems until they reached the marketable sizes of 100–140 snails/kg. At all stocking densities, final increments in mean length and weight of snails held in flow-through systems were higher than those in recirculating systems, there was a significant effect (p < 0.05) of culture system on final length and weight. Mean (±SE) survival of snails held at 100 ind. m^–2 in the flow-through system was 100.0 ± 0.1% but was not significantly higher than survival in any other treatment (p > 0.05). Mean survival of snails held in the recirculating system was not significantly lower than at any stocking density in the flow-through system (p > 0.05).     

Effect of combination feeding of <Emphasis Type="Italic">Nannochloropsis</Emphasis> and freshwater <Emphasis Type="Italic">Chlorella</Emphasis> on the fatty acid composition of rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> in a continuous culture

ABSTRACT:   A continuous culture of rotifer was conducted to investigate the effect of combination feeding of both a high density of Nannochloropsis oculata (N) and condensed freshwater Chlorella (FC) on the fatty acid composition of L-type rotifer Brachionus plicatilis in a continuous culture system. The algal feeding of the rotifers was carried out in three successive steps: N-feeding → N+FC-feeding → FC-feeding. The culture was conducted at 24°C and 25–27 psu in a 2000 mL bottle with 50% of water exchanged daily. The combination N+FC-feeding was effective in increasing rotifer density. The rotifers fed on N+FC (N+FC-R) had more non-polar lipids than polar ones, similar to those on N (N-R), opposite to the rotifers fed on FC (FC-R). N+FC-R contained higher levels of 16:2, 18:2n-6 (linoleic acid [LA]) and 20:2n-6, but lower levels of 18:1, 20:4n-6 (arachidonic acid), 20:5n-3 (eicosapentaenoic acid [EPA]) and 22:5n-3 (docosapentaenoic acid [DPA]) compared with N-R. Whereas N+FC-R contained higher levels of 16:1n-7, EPA and DPA, but lower levels of 16:2 and LA compared with FC-R. N+FC-R had more DPA in polar lipids than in non-polar ones. The Σn-6/Σn-3 ratio in N+FC-R was 0.9–1.0, significantly different from those in N-R (0.4) and FC-R (6.6–8.4). Therefore, it is inferred that the fatty acid profile of the N+FC-R cultured in a continuous culture system was affected by both N and FC. Also, the combination N+FC-feeding may be effective in manipulating the Σn-6/Σn-3 ratio in continuously cultured rotifers.     

Effect of <Emphasis Type="Italic">Cratoxylum formosum</Emphasis> on innate immune response and disease resistance against <Emphasis Type="Italic">Streptococcus agalactiae</Emphasis> in tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

The effect of aqueous extract of Cratoxylum formosum on innate immune response and disease resistance in tilapia Oreochromis niloticus was investigated. The fish were fed diets containing 0% (control), 0.5% (diet 1), 1% (diet 2), and 1.5% (diet 3) of C. formosum aqueous extract for 30 days. At the end of the experimental period, parameters of innate immune response including phagocytosis of blood leukocytes, lysozyme activity in plasma, and respiratory bust activity were examined. Feeding the fish with diet 2 and diet 3 for 20 days enhanced phagocytic activity and for 30 days stimulated lysozyme and respiratory burst activities. Diet 1 increased the phagocytic activity at 30 days, but did not affect the other measured parameters. All parameters were not significantly changed (P > 0.05) in the control group throughout the experiment. Following 30 days of feeding, fish were infected with S. agalactiae. The cumulative mortalities of bacterial-infected tilapia that were fed diet 1, diet 2, and diet 3 were 56, 12, and 10%, respectively, compared with 85% in the control group. These results indicate that the aqueous extract of C. formosum may elevate the innate immune response and enhance disease resistance in tilapia.     

Monitoring of <Emphasis Type="Italic">Francisella noatunensis</Emphasis> subsp. <Emphasis Type="Italic">orientalis</Emphasis> in farmed Nile tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>) in Brazil

Francisella noatunensis orientalis is a bacterium that causes emerging bacteriosis in Nile tilapia (Oreochromis niloticus) in many parts of the world, including Brazil. It is a non-motile, Gram-negative, strictly aerobic, facultative intracellular coccobacillus. This species of bacteria is responsible for low to high mortality in fish farms, causing economic losses for fish farmers. This study aimed to detect the presence of F. noatunensis orientalis using qPCR (real-time polymerase chain reaction) and to describe lesions caused by the bacterium in O. niloticus in Brazilian aquaculture. For this purpose, 360 fish from six fish farms (30 per farm) were sampled at two time points (n = 180 per sampling). Necropsies and histopathology were performed for lesion observation, in addition to qPCR and sequencing for detection and identification of Francisella species. Environmental data were collected using a multiparameter sonde YSI EXO2. All measured limnological variables were within the optimum range for cultivation of Nile tilapia. The major lesions present were melanization of the skin, splenomegaly, granulomas, and inflammatory cell responses. The prevalence of francisellosis varied from 0 to 86.66% between time periods and fish farms analyzed, and an outbreak was observed during the second sampling period. This study describes the prevalence of francisellosis in O. niloticus and reports that the lesions found are not exclusively associated with this bacterial disease.     

Characterization of <Emphasis Type="Italic">kiss2</Emphasis> and <Emphasis Type="Italic">kissr2</Emphasis> genes and the regulation of kisspeptin on the HPG axis in <Emphasis Type="Italic">Cynoglossus semilaevis</Emphasis>

Reproduction allows organisms to produce offspring. Animals shift from immature juveniles into mature adults and become capable of sexual reproduction during puberty, which culminates in the first spermiation and sperm hydration or ovulation. Reproduction is closely related to the precise control of the hypothalamic–pituitary–gonadal (HPG) axis. Kisspeptin peptides are considered as the important regulator of HPG axis in mammalian. However, the current understanding of kisspeptin in flatfish is not comprehensive. In this study, we cloned and analyzed the kiss2 and kissr2 genes in Cynoglossus semilaevis. Interesting alternative splicing in the 5′-untranslated regions (UTR) of the Cskissr2 gene was found. The expression profiles of Cskiss2 and Cskissr2 showed relative high messenger RNA (mRNA) levels at the late gastrula stage during embryonic development, at total length = 40 mm during early gonadal differentiation, and in the brains and gonads of all investigated tissues. These results suggested that the kisspeptin system participated in embryogenesis and in the regulation of gonadal differentiation and development. Considering that the control and regulatory mechanisms of kisspeptin in the central reproductive axis are still unclear, we documented that the intramuscular injection of kisspeptin caused different sGnRH and cGnRH mRNA levels in a dose- and tissue-dependent manner. The mRNA expressions of FSH and LH were stimulated in the ovary and were inhibited in the testis under the kisspeptin treatments. These results provided foundations for understanding the roles of kisspeptin in the neuroendocrine system in fish. The manipulation of the kisspeptin system may provide new opportunities to control the gonadal development and even reproduction in fish.     

Influence of <Emphasis Type="Italic">Tribulus terrestris</Emphasis> on testicular enzyme in fresh water ornamental fish <Emphasis Type="Italic">Poecilia latipinna</Emphasis>

The influence of Tribulus terrestris on the activities of testicular enzyme in Poecilia latipinna was assessed in lieu of reproductive manipulation. Different concentrations of (100, 150, 200, 250, and 300 mg) Tribulus terrestris extract and of a control were tested for testicular activity of enzymes in Poecilia latipinna for 2 months. The testis and liver were homogenized separately in 0.1 mol/l potassium phosphate buffer (0.1 mol/l, pH 7.2). The crude homogenate was centrifuged, and supernatant obtained was used as an enzyme extract for determination of activities. The activities of testicular functional enzyme ALP, ACP, SDH, LDH, and G6PDH levels were changed to different extent in treated groups compared with that of the control. The total body weight and testis weight were increased with the Tribulus terrestris-treated fish (Poecilia latipinna). These results suggest that Tribulus terrestris induced the testicular enzyme activity that may aid in the male reproductive functions. It is discernible from the present study that Tribulus terrestris has the inducing effect on reproductive system of Poecilia latipinna.     

Growth and survival of grouper <Emphasis Type="Italic">Epinephelus</Emphasis><Emphasis Type="Italic">coioides</Emphasis> (Hamilton) larvae fed free-living nematode <Emphasis Type="Italic">Panagrellus redivivus</Emphasis> at first feeding

The free-living nematode, Panagrellus redivivus, was tested as live food for grouper Epinephelus coioides larvae during the first feeding stage. A series of experiments were conducted to determine the acceptability of the free-living nematodes in grouper larvae at first feeding, the optimum nematode density and the response of the larvae to nutritionally enriched nematode. All experiments were conducted in 200-L conical tanks filled with 150-L filtered seawater and stocked at 15 larvae L⁻¹. Duration of feeding experiments was up to day 21 (experiment 1) and 14 days (experiment 2 and 3). Brachionus plicatilis and Artemia (experiment 1) and Brachionus plicatilis alone (experiment 2 & 3) was used as the control treatment. Observations indicated that the grouper larvae readily fed on free-living nematodes as early as 3 days posthatching, the start of exogenous feeding. Optimum feeding density for the larvae was 75 nematodes ml⁻¹. The enrichment of cod liver oil or sunflower oil influenced the total lipids and n-3 highly unsaturated fatty acids of P. redivivus, which in turn influenced those of the grouper larvae, however, growth and survival of the larvae were not affected (P > 0.05). The results from this investigation showed that the nematode, P. redivivus, can be used as first live food for grouper larvae from the onset of exogenous feeding until they could feed on Artemia nauplii.     

Effects of dietary microencapsulated <Emphasis Type="Italic">Cymbopogon flexuosus</Emphasis> essential oil on reproductive-related parameters in male <Emphasis Type="Italic">Rhamdia quelen</Emphasis>

In aquaculture, nutrition and supplemented diets have been shown to affect broodstock reproductive performance. In this study, we investigated the effects of dietary supplementation with Cymbopogon flexuosus essential oil (CFEO) microcapsules on reproductive-related parameters in silver catfish (Rhamdia quelen) male broodfish. Adult male broodstocks were separated into three groups according to the concentrations of supplemented CFEO (0.0?=?control; 1.0 or 3.0 mL per kg of diet). After 20 days under experimental conditions, the animals were euthanized and the gonads were harvested for gonadosomatic index, sperm analysis, oxidative stress, and histopathology; testosterone levels were measured in the plasma; gene expression of prl, smtl, pomca, and pomcb was assessed in the pituitary gland by real-time PCR. The results showed no alterations on reproductive parameters in R. quelen males treated with Cymbopogon flexuosus essential oil compared to the control-diet animals. In conclusion, CFEO microcapsules supplied for 20 days in the concentrations of 1.00 or 3.00 mL per kilogram of diet did not affect the reproduction criteria evaluated in this study in male silver catfish.     

Recent expansion of Northeast Atlantic and Mediterranean populations of <Emphasis Type="Italic">Melicertus</Emphasis> (<Emphasis Type="Italic">Penaeus</Emphasis>) <Emphasis Type="Italic">kerathurus</Emphasis> (Crustacea: Decapoda)

We analysed the genetic diversity of Melicertus kerathurus (Penaeidae), a commercially valuable penaeid shrimp that is distributed in the Mediterranean Sea and eastern Atlantic Ocean. We examined the polymorphism of a 494 bp DNA segment of the mitochondrial COI region in 173 individuals, sampled in nine Mediterranean and two Atlantic samples, covering the whole range of the species from the tropical waters of the Gulf of Guinea to the eastern part of the Mediterranean Sea. The mean nucleotide and haplotype diversities were π = 0.00275 and h = 0.718, respectively, for the global data set, with the highest values occurring in the African samples and the lowest in the Adriatic Sea. A clear sample differentiation was found (F _st = 0.194), but this did not reflect a geographical pattern and there were only faint traces of an Atlantic–Mediterranean subdivision. Mismatch analysis and a high significant negative value of Tajima’s D suggested that M. kerathurus is not at mutation drift-equilibrium, but underwent a recent expansion after a period of low effective sample size. A postglacial recolonisation of the Mediterranean from an Atlantic refuge could be hypothesised based on these data.     

Random sequencing of genomic DNA of <Emphasis Type="Italic">Photobacterium damselae</Emphasis> subsp. <Emphasis Type="Italic">piscicida</Emphasis>

ABSTRACT:   Random sequencing of the genomic DNA of Photobacterium damselae subsp. piscicida was conducted. The sequences were assembled into 930 contigs. The total length of these contigs accounts for 37.5% of the genome of P. damselae subsp. piscicida. The contigs contain 2055 open reading frames (ORFs) that have homology with genes in the GenBank database. Furthermore, some of the ORFs have homology with reported virulence related genes, and are classified as encoding colonization factors, exotoxin, and lipopolysaccharide (LPS). Thirty-seven ORFs have homology with colonization factors. In those colonization factors, 27, three, and four ORFs have homology with polar flagellar-related genes, capsule-related genes, and others (accessory colonization factors, superoxide dismutase (Cu-Zn), MshA biogenesis protein, and heme receptor genes), respectively. Five ORFs have homology with exotoxin-related genes (2 hemolysin, phospholipase, hyaluronidase, lysophospholipase L2 genes). Further, six ORFs have homology with LPS-related genes.     

Effects of potential probiotic <Emphasis Type="Italic">Lactococcus lactis</Emphasis> subsp. <Emphasis Type="Italic">lactis</Emphasis> on digestive enzymatic activities of live feed <Emphasis Type="Italic">Artemia franciscana</Emphasis>

In the present study, cellulase, protease, lipase and amylase activities were performed to investigate the effects of Lactococcus lactis subsp. lactis CF4MRS bioencapsulation of Artemia franciscana. Our results show that cellulase activities (total cellulase—FPase activity, exoglucanase and endoglucanase—CMCase activity) were significantly higher (P < 0.05) in A. franciscana tissue homogenates compared to those in the control group after 8 h of L. lactic bioencapsulation. Notably, an exception case was found in β-D-glucosidase activity, whereby the cellulase activity was not significantly different (P > 0.05) compared to the control group. Administrations of L. lactis at cell concentration of 10⁸ CFU mL^?1 showed considerable improvement of other important enzymatic activities such as amylase, protease and lipase in A. franciscana. The amylase/protease ratio in probiotic-treated A. franciscana was recorded at 0.343, approximately two times higher than those without probiotic administration (0.184). In contrary, amylase/lipase ratio showed half of a reduction (0.330) in L. lactis-administrated A. franciscana compared to the control (0.614). Our study suggests that important digestive enzymes, e.g., cellulase, amylase, protease and lipase, can be enhanced through bioencapsulation of A. franciscana with L. lactis subsp. lactis, which could in turn lead to further stimulation of endogenous enzymes in the fish and shrimp larvae.