Supplementation of heat‐inactivated Bacillus clausii DE5 in diets for grouper,Epinephelus coioides,improves feed utilization,intestinal and systemic immune responses and not growth performance

In recent years, more and more attentions have been paid to the development and application of probiotics in aquaculture, and viable probiotics have been extensively studied, while rare information was available about inactivated probiotics in aquaculture. Therefore, in this study, a feeding trial was designed to investigate the effect of dietary supplementation of heat‐inactivated probiotic Bacillus clausii DE5 on growth performance, immune response and key immune genes expression in head kidney and intestine in grouper Epinephelus coioides. Fish were fed for 60 days with control diet (C) and two experimental diets containing 1.0 × 10⁸ CFU/g live (T1) and heat‐inactivated (T2) B. clausii DE5, respectively. The probiotic treatments did not affect the final weight, weight gain (WG) and specific growth rate (SGR) of E. coioides at days 30 and 60 (p > .05), while both heat‐inactivated and live B. clausii DE5 significantly decreased the feed intake and feed conversion ratio (FCR) at day 60 (p < .05). Serum lysozyme activity and complement C3 level in the two probiotic treatments were significantly higher than those in the control (p < .05). The lysozyme activity and complement C3 level at day 60 were significantly higher than those at day 30 (p < .05), while no significant interaction effect between diet and administration date was observed. Moreover, the heat‐inactivated B. clausii DE5 significantly improved the expression of TLR5, pro‐inflammatory cytokines (IL‐8 and IL‐1β) and TGF‐β1 in head kidney and intestine (p < .05), while the live probiotic did not show any significant effect on the expression of these key immune‐related genes in head kidney and intestine. These results indicate that dietary supplementation of heat‐inactivated B. clausii DE5 effectively improved feed utilization and both the local and systemic immune responses of E. coioides.     

Effects of supplemental phytic acid on the apparent digestibility and utilization of dietary amino acids and minerals in juvenile grass carp (Ctenopharyngodon idellus)

A feeding trial was conducted for 8 weeks to evaluate the effects of supplemental phytic acid (PA) on the apparent digestibility and utilization of dietary amino acids (AAs) and minerals in juvenile grass carp. Five experimental diets consisted of graded levels of PA (0.2, 4.7, 9.5, 19.1 and 38.3 g kg^?1, named as P0, P5, P10, P20 and P40). Triplicate groups of fish (initial weight, 22.37 ± 0.16 g) were fed twice daily (08:00 and 16:00 h). The crude protein content in whole body significantly (p < .05) decreased in fish fed with 19.1 and 38.3 g PA kg^?1 diet. Supplemental PA (>4.7 g kg^?1) significantly reduced the apparent digestibility coefficient (ADC) of AAs (Asp, Thr, Ser, Glu, Gly, Ala, Cys, Val, Met, Ile, Leu, Phe, Lys, Pro, His and Arg) and the ADC of minerals (P, Ca, Mg, Zn, Cu, Fe and Mn) in grass carp. The contents of minerals (P, Ca, Mg and Zn) in whole body and bone were also found to be significantly reduced in dietary PA > 4.7 g kg^?1, while the bone ash, serum Alkp and Zn contents were found to be significantly decreased when the PA supplementation level was above 9.5 g kg^?1, and the contents of serum Ca and Mg were found to be markedly altered in higher PA‐supplemented groups. The results indicated that supplemental PA decreased the apparent digestibility and utilization of AAs and minerals, and thus reduced the feed utilization of grass carp, suggesting that the level of total PA should be below 4.7 g kg^?1 in grass carp diet.     

Successful rearing of whiteleg shrimp Litopenaeus vannamei larvae fed a desiccation‐tolerant nematode to replace Artemia

The nematode Panagrolaimus sp. was tested as live feed to replace Artemia nauplii during first larval stages of whiteleg shrimp Litopenaeus vannamei. In Trial 1, shrimp larvae were fed one of four diets from Zoea 2 to Postlarva 1 (PL1): (A) Artemia nauplii, control treatment; (NC) nematodes enriched in docosahexaenoic acid (DHA) provided by the dinoflagellate Crypthecodinium cohnii; (N) non‐enriched nematodes; and (Algae) a mixture of microalgae supplemented in C. cohnii cells. In Trial 2, shrimp were fed (A), (NC) and a different treatment (NS) with nematodes enriched in polyunsaturated fatty acids (PUFAs) provided by the commercial product S.presso^®, until Postlarva 6 (PL6). Mysis 1 larvae fed nematodes of the three dietary treatments were 300 μm longer (3.2 ± 0.3 mm) than control larvae. At PL1, control shrimp were 300 μm longer (4.5 ± 0.3 mm) than those fed DHA‐enriched or PUFAs‐enriched nematodes. No differences were observed in length and survival at PL6 between control larvae and those fed DHA‐enriched nematodes (5.1 ± 0.5 mm; 33.1%–44.4%). Shrimp fed microalgae showed a delay in development at PL1. This work is the first demonstration of Panagrolaimus sp. suitability as a complete substitute for Artemia in rearing shrimp from Zoea 2 to PL6.     

Effect of ingredient particle sizes and dietary viscosity on digestion and faecal waste of striped catfish (Pangasianodon hypophthalmus)

The ingredients’ particle size and dietary viscosity may alter digestion, performance and faecal waste management of fish. This study aimed to assess the effect of grinding screen sizes of feed ingredients and dietary viscosity on digestibility, faecal waste and performance of striped catfish (Pangasianodon hypophthalmus, Sauvage, 1878). The experiment had a 2 × 3 factorial‐design: two feed mesh particle sizes, by grinding ingredient mixtures at two screen sizes (0.8 versus 1.0 mm); and three dietary viscosity levels, created by exchanging carboxymethylcellulose by guar gum (GG) (0, 3 and 6 g of GG/kg of diet). Six diets were assigned to 18 tanks, each connected to three faecal settling tanks. All aquaria were stocked with 20 fish (82 g per fish). After 52 experimental days, dietary viscosity negatively affected both feed digestibility and performance of striped catfish; as a result, the amount of organic matter in the culture system through faeces had increased significantly. The coarse diets significantly increased the digestibility of dry matter and carbohydrate but worsened feed conversion ratio. Increasing dietary viscosity tended to increase the viscosity and moisture content of the faeces, but significantly accelerated the faecal disintegration through the reduction of both faecal recovery and the amount of recovered faeces.     

First approach to studying the genetics of the meagre (Argyrosomus regius; Asso, 1801) using three multigene families

The meagre (Argyrosomus regius) is a commercially important species for fisheries, and aquaculture production has been increasing in recent years. However, this growing importance has not been matched by increased genetic knowledge of the species. For this reason, an initial approach has been made to understanding the genetics of the meagre, using three particular multigene families (45S and 5S ribosomal DNA and U2 snRNA gene) as both molecular and cytogenetic markers. Only one type of 5S rDNA and only one ITS‐1 (from 45S rDNA) were obtained, and these therefore could provide the basis for the development of genetic markers for this species. However, the U2 snRNA gene produced a multi‐band electrophoretic pattern, and some of these bands were demonstrated to be linked with the U1 snRNA gene. This linkage could also provide a good species‐specific marker which could be useful for tracing the evolutionary history of the Sciaenidae family. In this study, we have described for the first time the karyotype of the meagre: this is composed of 48 acrocentric chromosomes. Each of the three gene families were localized on different chromosome pairs, although the U2 snRNA gene probe was also located scattered throughout the genome of the meagre, as expected by the amplification pattern.     

Apparent digestibility coefficients of selected feed ingredients for juvenile snakehead,Ophiocephalus argus

Apparent digestibility coefficients (ADCs) of dry matter, crude protein, crude lipid, gross energy, phosphorus and amino acids in Peruvian fish meal, poultry by‐product meal, meat and bone meal, spray‐dried blood meal, hydrolysed feather meal, corn gluten meal, soybean meal, peanut meal, cottonseed meal and rapeseed meal were determined for juvenile snakehead (Ophiocephalus argus) with initial mean body weight of 78.1 g. A reference diet and test diets that consisted of a 70 : 30 mixture of the reference diet to test ingredient were used with 5 g kg^?1 Cr₂O₃ as an external indicator. Fish meal, poultry by‐product meal and corn gluten meal had higher ADCs of dry matter, crude protein, and gross energy among ingredients tested. Dry matter ADCs ranged 61.9–81.5% for animal ingredients and corn gluten meal and ranged 52.2–68.0% for soybean meal, peanut meal, cottonseed meal and rapeseed meal. Energy ADCs of ingredients followed similar trends to differences in dry matter digestibility. Protein ADCs of animal and plant ingredients ranged 73.6–92.8% and 75.3–85.6%, respectively. Amino acid ADCs generally reflected protein digestibility. Lipid ADCs were relatively high for the ingredients tested. Phosphorus ADCs of animal and plant ingredients ranged 39.5–65.2% and 38.7–57.1%, respectively.     

Evaluations of Hilyses™, fermented Saccharomyces cerevisiae,on rainbow trout (Oncorhynchus mykiss) growth performance,enzymatic activities and gastrointestinal structure

This study examined the effects of Hilyses?, fermented Saccharomyces cerevisiae (S. cerevisiae), on rainbow trout growth performance, haematological parameters, digestive enzyme activities and gastrointestinal structure. Rainbow trout (mean weight 100–110 g) were fed dietary Hilyses? (5 g kg^?1) and control diet without Hilyses? for 50 days. Results of this study demonstrated that yeast supplementation in treatment group resulted in increased feed intake, followed by improved feed conversion ratio (FCR) and growth performance. Significant increases were also observed in trypsin and amylase activities in juvenile fish fed treatment diet. Light microscopy demonstrated that both groups of fish displayed normal morphology of proximal intestine and pyloric caeca. In yeast‐treated group, higher density of the goblet cells per villus in the proximal intestine was shown. No effects on haematological parameters and carcass chemical composition were noted. It is therefore possible to use fermented S. cerevisiae supplementation to significantly improve the gastrointestinal structure and growth performance in rainbow trout.     

Effects of Neutral Phytase Supplementation on Biochemical Parameters in Grass Carp,Ctenopharyngodon idellus,and Gibel Carp,Carassius auratus gibelio,Fed Different Levels of Monocalcium Phosphate

Two 8‐wk studies were conducted to evaluate the effects of neutral phytase supplementation on hemato‐biochemical status, liver biochemical parameter, and intestinal digestive enzyme activity of grass carp, Ctenopharyngodon idellus, and gibel carp, Carassius auratus gibelio, fed with different levels of monocalcium phosphate (MCP). The control diet was prepared with 2% MCP but without phytase (P2.0). The other three experimental diets were prepared with the addition of 1.5, 1.0, and 0.5% MCP, respectively, when supplemented with 500 U/kg neutral phytase in each diet and designated as PP1.5, PP1.0, and PP0.5, respectively. The results indicated that the serum alkaline phosphatase (ALP), alanine transaminase (ALT), and aspartate transaminase (AST) activities, as well as the albumin (ALB) content were increased in grass carp (P < 0.05) and gibel carp (P > 0.05) fed with phytase‐supplemented diets. Meanwhile, the serum cholesterol, high‐density lipoprotein, and total protein contents of the two species of fish were increased in comparison to the control. In addition, dietary phytase inclusion did not significantly affect hepatic ALP, ALT, and AST activities in the two species of carp fed with different levels of MCP. Amylase activity increased in foregut and hindgut of both species when fed with the phytase‐supplemented diets while lipase activity was reduced in the foregut and hindgut in both fish. This study suggests that neutral phytase supplementation increases serum ALP, ALT, and AST activities but does not notably affect these enzyme activities in the liver of the two species of carp when fed different levels of MCP. On the other hand, amylase activity increased while lipase activity was reduced in the intestine of the species of carp fed with phytase‐supplemented diets.     

Progeny Production of the Copepods Pseudodiaptomus euryhalinus and Tisbe monozota in Monospecific and Mixed Cultures

The objective of this research was to culture the calanoid copepod Pseudodiaptomus euryhalinus, Johnson 1939, and the harpacticoid Tisbe monozota, Bowman, 1962, in monospecific and combined cultures (P. euryhalinus : T. monozota at 1:1, 2:1, and 1:2 starting ratios), and to compare the nauplii, copepodite, and adult production. Mean total production ranged from 740.3 ± 137.5 organisms/L (P. euryhalinus) to 884.3 ± 489.7 organisms/L (T. monozota) for monospecific cultures. The 1:1 ratio mixed cultures gave 780.4 ± 155.8 organisms/L, those with the 2:1 and 1:2 P. euryhalinus and T. monozota starting ratios produced 710.1 ± 195.2 and 799.7 ± 232.5 organisms/L, respectively, and there were no significant difference among treatments. All mixed cultures gave significantly lower copepodite productions than the monocultures of each species. In addition, the tendency to a decreased progeny production of the initial females of T. monozota indicates that the outcome of long‐term mixed cultures would production either a high dominance of P. euryhalinus, or monocultures of this species .