Dietary astaxanthin augments disease resistance of Asian seabass, Lates calcarifer (Bloch, 1790), against Vibrio alginolyticus infection.

This investigation describes the impacts of dietary provisioning with astaxanthin on hemato-biochemistry, non-specific immunity, and disease resistance of the Asian seabass, Lates calcarifer, against the virulent Vibrio alginolyticus; with specific reference to dose-response associations and variations over different post-infection periods (0-, 7-, and 14-day). Triplicate groups of fish weighing 28 g, on average, were fed various diets (C, the control or astaxanthin-free; AXT50, 50 mg astaxanthin kg-1 diet; AXT100, 100 mg astaxanthin kg-1 diet; and AXT150, 150 mg astaxanthin kg-1 diet) for 90 days and subsequently challenged with V. alginolyticus at the end of the feeding period. Experimental infection unveiled that supplemented fish demonstrated significant improvements (P < 0.05) of hematological parameters (white blood cell [WBC] and red blood cell [RBC] counts, and hemoglobin and hematocrit levels) when fed diets with elevating supplemental doses of astaxanthin through distinct post-infection periods (0-, 7-, and 14-day). Furthermore, the administration of dietary astaxanthin at escalating levels markedly enhanced (P < 0.05) the serum biochemical profile (aspartate aminotransferase [AST], alanine aminotransferase [ALT], glucose, cortisol, cholesterol, and triglyceride contents) of challenged fish, resulting in better welfare. Significantly higher (P < 0.05) contents of serum total protein were observed in supplemented fish, as opposed to the control. Additionally, immunological defense mechanisms (lysozyme activity, phagocytic activity, respiratory burst activity, and total serum immunoglobulin) of challenged fish were pronouncedly elicited (P < 0.05) following the ingestion of astaxanthin. Besides, the supplementation with dietary astaxanthin significantly augmented (P < 0.05) the post-challenge survival rate of fish. Collectively, the results manifest that supplementary feeding of astaxanthin is effective in reinforcing fish immunocompetence and disease resistance against V. alginolyticus infection.

Data on PCR primer design for glucose 6-phosphate dehydrogenase gene and the effects of dietary carbohydrate levels on its expression in the liver of Malaysian mahseer (Tor tambroides).

The enzyme glucose-6-phosphate dehydrogenase (G6PD) catalyses the metabolite glucose-6-phosphate in producing NADPH during the first phase of pentose-phosphate pathway thus provides reducing power to all cells for cellular growth, antioxidant defence, and biosynthetic reactions in all living organism. The deliberate inclusion of starch as carbohydrate source in commercial feed however may affect the G6PD hepatic activity in cultured fish. We designed a set of primers to target G6PD gene in the popular Malaysian aquaculture species, Tor tambroides. For this dataset, the molecular characteristics of obtained T. tambroides G6PD (TtG6PD) nucleotide sequence was analysed using multiple alignments and phylogenetic analyses of the deduced amino acids. The set of primers obtained were then used in a study to evaluate the effect of different dietary carbohydrate inclusion levels on the hepatic TtG6PD mRNA expression of the T. tambroides fingerlings. Four groups of fish were given a dietary treatment of 15%, 20%, 25% and 30% starch at the optimal inclusion level of 23.4% for 10 weeks. The TtG6PD mRNA transcripts were measured using real-time-PCR assays and its expression normalized against ß-actin, which acts as the internal control gene. This article provides supportive data in relation between hepatic TtG6PD mRNA gene expression in T. tambroides and how it is influenced by its dietary carbohydrate intake. These data will also assist in further nutritional genomic studies of carbohydrate and energy utilization for all species in the mahseer family.

Effect of temperature on thermophilic composting of aquaculture sludge: NH3 recovery, nitrogen mass balance, and microbial community dynamics.

Development of thermophilic composting for maximizing NH3 gas recovery would enable the production of a nitrogen source which is free from pathogen/heavy metal, for the cultivation of high-value microalgae. The present study examined the effect of NH3 recovery, nitrogen mass balance, and microbial community dynamics on thermophilic composting of shrimp aquaculture sludge. The emission of NH3 gas at 60 and 70 °C was 14.7% and 15.6%, respectively, which was higher than that at 50 °C (9.0%). The nitrogen mass balance analysis revealed that higher temperatures enhanced the solubilization of non-dissolved nitrogen and liberation of NH3 gas from the produced NH4+-N. High-throughput microbial community analysis revealed the shift of the dominant bacterial group from Bacillus to Geobacillus with the rise of composting temperature. In conclusion, thermophilic composting of shrimp aquaculture sludge at 60-70 °C was the most favorable condition for enhancing NH3 gas recovery.

Improvement of feed pellet characteristics by dietary pre-gelatinized starch and their subsequent effects on growth and physiology in tilapia.

A 9-week study was conducted to compare dietary corn starch (CS) or tapioca starch (TS), with or without being pre-gelatinized (PG), on the growth, feeding efficiencies, plasma and muscle biochemistry, intestinal short chain fatty acids (SCFA), and liver glycogen of triplicate groups of 20 red hybrid tilapia (Orecohromis sp.). Various pellet characteristics were evaluated, along with their surface and cross sectional microstructure. The PG diets had significantly higher water stability, bulk density, and protein solubility, along with a smoother surface. Tilapia fed the TS diet had lower growth than had all other tilapia, but were significantly improved when diet was pre-gelatinized. In the PG treatments, intestinal SCFA significantly decreased while plasma glucose, cholesterol and triglycerides, as well as liver glycogen, significantly increased. Fish fed the CS diet had significantly more long chain polyunsaturated fatty acid than had those fed by other treatments. Pre-gelatinization may improve fish productivity and offer greater flexibility during aquafeed production.

Population Structure of the Blue Swimmer Crab Portunus pelagicus in Coastal Areas of Malaysia Inferred from Microsatellites.

Chuan Jian Chai, Yuzine Bin Esa, Muhammad Fadhil Syukri Ismail, and Mohd. Salleh Kamarudin (2017) Portunus pelagicus, distributed throughout the Indo-West Paci c region, is one of the large and edible species of blue swimmer crabs. Increasing demand for the frozen and canned crabmeat industry worldwide has now relied mainly on P. pelagicus which in turn generates splendid income for the fisherman communities. In the present study, the population genetic structure of P. pelagicus was examined using six pairs of microsatellite loci. A total of 87 crab samples were collected from five different coastal areas of Malaysia. Genomic DNA was extracted from each sample for polymerase chain reaction (PCR) amplification and fragment analysis. Four out of six microsatellite primers revealed polymorphic loci in P. pelagicus sampled. The number of alleles per locus in P. pelagicus ranged from 14 to 34. Microsatellites analyses indicated low levels of genetic differentiation among the P. pelagicus populations. The average observed heterozygosity (HO = 0.48) obtained was lower than the standard heterozygosity found in most marine populations (H O = 0.79). The high F IS values (mean F IS = 0.4756) and low F ST values (mean F ST = 0.0413) also suggested the existence of inbreeding among different populations of P. pelagicus. In conclusion, this study was able to shed light on the population structure of P. pelagicus in coastal areas of Malaysia.

Trypsin-modulating oostatic factor (TMOF) decreased the survival, growth and digestion enzymes of Macrobrachium rosenbergii.

Trypsin-modulating oostatic factor (TMOF) is an effective mosquito larvicide, but information on its potential toxicity to non-target organisms is limited. To investigate this, triplicate groups of 10 Macrobrachium rosenbergii were exposed to 0, 10, 50 or 100 mg/L nominal TMOF concentrations for 12 days. Tail moisture, crude protein, and hepatopancreatic glycogen/histopathology were unaffected, but increasing TMOF linearly decreased survival and growth. TMOF at the lowest concentration employed significantly decreased trypsin and chymotrypsin activities.

Effects of varying dietary carbohydrate levels on growth performance, body composition and liver histology of Malaysian mahseer fingerlings (Tor tambroides).

We investigated the effects of four iso-nitrogenous (40% crude protein) and iso-caloric (17.6 kJ g(-1)) diets with different dietary carbohydrate levels (15%, 20%, 25% and 30%) on the growth performance, feed utilization efficiency, body composition and liver histology of Malaysian mahseer (Tor tambroides) fingerlings in a 10-week feeding trial. Fish (initial weight of 0.8?0.1 g; initial total length 4.2?0.1 cm) were fed twice daily at 4% body mass. Dietary carbohydrate level had significant effects (P<0.05) on weight gain, SGR (specific growth rate), FCR (feed conversion rate), PER (protein efficiency rate), survival percentage and all nutrient retention values (PRV, LRV, CRV, ERV). Protein, carbohydrate and gross energy composition of the fish body were also significantly differed (P<0.05) among treatments. Liver histology showed mild hepatic steatosis and hypertrophy for fishes receiving a higher dietary carbohydrate inclusion. In general, treatments with 20% and 25% dietary carbohydrate levels produced better growth results compared to the rest of the treatments. Using a second-order polynomial regression analysis model, the optimal dietary carbohydrate level of 23.4% was estimated for mahseer fingerlings. ?

Dietary lipid requirement of lemon fin barb hybrid.

The present study has been conducted to determine the optimal dietary requirement lipid of lemon fin barb hybrid fingerlings. Five isonitrogenous diets (35% protein) with graded lipid levels of 0%, 4%, 6%, 8% and 12% were fed to 225 lemon fin barb hybrid fingerlings (4.3 ? 0.5 g), which were randomly and equally distributed into twelve 60 l aquaria. The diets were randomly assigned to the aquaria in triplicates, and the feeding experiment was conducted for 10 weeks. Fish were fed at 5% body weight and the feeding ration was adjusted fortnightly. The weight gain, specific growth rate (SGR), protein efficiency ratio (PER) and feed conversion ratio (FCR) of fish fed 4% dietary lipid were significantly superior (P<0.05) than those fed with other diets, except for 6% lipid. Except for whole body moisture content, there were significant differences (P<0.05) between whole body proximate composition and nutrient retentions of fish fed with different test diets. The results showed that palmitic acid was the main saturated fatty acid (SFA), oleic acid was the predominant MUFA while arachidic acid and docosahexanoic were the major PUFAs in the muscle and liver tissue of fish. The n-3/n-6 ratio ranged from 0.66 to 2.65. From the results of this study, a dietary lipid of 4% was recommended for lemon fin barb hybrid. ?

Administration of Bacillus subtilis strains in the rearing water enhances the water quality, growth performance, immune response, and resistance against Vibrio harveyi infection in juvenile white shrimp, Litopenaeus vannamei.

In this study, vegetative cell suspensions of two Bacillus subtilis strains, L10 and G1 in equal proportions, was administered at two different doses 10(5) (BM5) and 10(8) (BM8) CFU ml(-1) in the rearing water of shrimp (Litopenaeus vannamei) for eight weeks. Both probiotic groups showed a significant reduction of ammonia, nitrite and nitrate ions under in vitro and in vivo conditions. In comparison to untreated control group, final weight, weight gain, specific growth rate (SGR), food conversion ratio (FCR) and digestive enzymatic activity were significantly greater in the BM5 and BM8 groups. Significant differences for survival were recorded in the BM8 group as compared to the control. Eight weeks after the start of experiment, shrimp were challenged with Vibrio harveyi. Statistical analysis revealed significant differences in shrimp survival between probiotic and control groups. Cumulative mortality of the control group was 80%, whereas cumulative mortality of the shrimp that had been given probiotics was 36.7% with MB8 and 50% with MB5. Subsequently, real-time RT-PCR was employed to determine the mRNA levels of prophenoloxidase (proPO), peroxinectin (PE), lipopolysaccharide- and ß-1,3-glucan- binding protein (LGBP) and serine protein (SP). The expression of all immune-related genes studied was only significantly up-regulated in the BM5 group compared to the BM8 and control groups. These results suggest that administration of B. subtilis strains in the rearing water confers beneficial effects for shrimp aquaculture, considering water quality, growth performance, digestive enzymatic activity, immune response and disease resistance.

Effects of Bacillus subtilis on the growth performance, digestive enzymes, immune gene expression and disease resistance of white shrimp, Litopenaeus vannamei.

We studied the effect of two probiotic Bacillus subtilis strains on the growth performance, digestive enzyme activity, immune gene expression and disease resistance of juvenile white shrimp (Litopenaeus vannamei). A mixture of two probiotic strains, L10 and G1 in equal proportions, was administered at two different doses 10(5) (BM5) and 10(8) (BM8) CFU g(-1) feed to shrimp for eight weeks. In comparison to untreated control group, final weight, weight gain and digestive enzyme activity were significantly greater in shrimp fed BM5 and BM8 diets. Significant differences for specific growth rate (SGR) and survival were recorded in shrimp fed BM8 diet as compared with the control; however, no significant differences were recorded for food conversion ratio (FCR) among all the experimental groups. Eight weeks after the start of the feeding period, shrimp were challenged with Vibrio harveyi. Statistical analysis revealed significant differences in shrimp survival between probiotic and control groups. Cumulative mortality of the control group was 63.3%, whereas cumulative mortality of the shrimp that had been given probiotics was 20.0% with BM8 and 33.3% with BM5. Subsequently, real-time PCR was employed to determine the mRNA levels of prophenoloxidase (proPO), peroxinectin (PE), lipopolysaccharide- and ß-1,3-glucan-binding protein (LGBP) and serine protein (SP). The expression of all immune-related genes studied was significantly up-regulated (P < 0.05) in the shrimp fed BM5 and BM8 diets compared to the control group. These findings demonstrate that administration of B. subtilis strains, L10 and G1, can improve growth performance and disease resistance through an enhanced immune response in shrimp.

Selection and identification of non-pathogenic bacteria isolated from fermented pickles with antagonistic properties against two shrimp pathogens.

In this study, potential probiotic strains were isolated from fermented pickles based on antagonistic activity against two shrimp pathogens (Vibrio harveyi and Vibrio parahaemolyticus). Two strains L10 and G1 were identified by biochemical tests, followed by16S ribosomal RNA gene sequence analysis as Bacillus subtilis, and characterized by PCR amplification of repetitive bacterial DNA elements (Rep-PCR). Subsequently, B. subtilis L10 and G1 strains were tested for antibacterial activity under different physical conditions, including culture medium, salinity, pH and temperature using the agar well diffusion assay. Among the different culture media, LB broth was the most suitable medium for antibacterial production. Both strains showed the highest level of antibacterial activity against two pathogens at 30 °C and 1.0% NaCl. Under the pH conditions, strain G1 showed the greatest activity against V. harveyi at pH 7.3-8.0 and against V. parahaemolyticus at pH 6.0-8.0, whereas strain L10 showed the greatest activity against two pathogens at pH 7.3. The cell-free supernatants of both strains were treated with four different enzymes in order to characterize the antibacterial substances against V. harveyi. The result showed considerable reduction of antibacterial activity for both strains, indicating the proteinaceous nature of the antibacterial substances. A wide range of tolerance to NaCl, pH and temperature was also recorded for both strains. In addition, both strains showed no virulence effect in juvenile shrimp Litopenaeus vannamei. On the basis of these results and safety of strains to L. vannamei, they may be considered for future challenge experiments in shrimp as a very promising alternative to the use of antibiotics.

Optimum dietary protein requirement of Malaysian mahseer (Tor tambroides) fingerling.

The optimum dietary protein requirement of the Malaysian mahseer (Tor tambroides) fingerlings was determined in this study. In this completely randomized designed experiment, formulated diets of five levels of dietary protein (30, 35, 40, 45 and 50%) were tested on the T. tambroides fingerlings (initial body weight of 5.85 +/- 0.40 g), reared in aquarium fitted with a biofiltering system. The fingerlings were fed twice daily at 5% of biomass. The fingerling body weight and total length was taken at every two weeks. Mortality was recorded daily. The dietary protein had significant effects on the body weight gain and Specific Growth Rate (SGR) of the fingerlings. The body weight gain and SGR of fingerlings fed with the diet with the dietary protein level of 40% was significantly higher (p<0.05) than that of 30, 35 and 50%. The feed conversion ratio of the 40% dietary protein was the significantly lowest at 2.19 +/- 0.163. The dietary protein level of 40% was the most optimum for T. tambroides fingerlings.

Anthropogenic impacts on heavy metal concentrations in the coastal sediments of Dumai, Indonesia.

Concentrations of Cd, Cu, Pb, Zn, Ni and Fe were determined in the surface sediments to investigate the distributions, concentrations and the pollution status of heavy metals in Dumai coastal waters. Sediment samples from 23 stations, representing 5 different site groups of eastern, central and western Dumai and southern and northern Rupat Island, were collected in May 2005. The results showed that heavy metal concentrations (in microg/g dry weight; Fe in %) were 0.88 (0.46-1.89); 6.08 (1.61-13.84); 32.34 (14.63-84.90); 53.89 (31.49-87.11); 11.48 (7.26-19.97) and 3.01 (2.10-3.92) for Cd, Cu, Pb, Zn, Ni and Fe, respectively. Generally, metal concentrations in the coastal sediments near Dumai city center (eastern and central Dumai) which have more anthropogenic activities were higher than those at other stations. Average concentration of Cd in the eastern Dumai was slightly higher than effective range low (ERL) but still below effective range medium (ERM) value established by Long et al. (Environmental Management 19(1):81-97, 1995; Environmental Toxicology Chemistry 17(4):714-727, 1997). All other metals were still below ERL and ERM. Calculated enrichment factor (EF), especially for Cd and Pb, and the Pollution load index (PLI) value in the eastern Dumai were also higher than other sites. Cd showed higher EF when compared to other metals. Geo-accumulation indices (I(geo)) in most of the stations (all site groups) were categorized as class 1 (unpolluted to moderately polluted environment) and only Cd in Cargo Port was in class 2 (moderately polluted). Heavy metal concentrations found in the present study were comparable to other regions of the world and based on the calculated indices it can be classified as unpolluted to moderately polluted coastal environment.