Delivery of HUFA, probionts and biomedicine through bioencapsulated Artemia as a means to enhance the growth and survival and reduce the pathogenesity in shrimp Penaeus monodon postlarvae

One of the major problems in the shrimp culture industry is the difficulty in producing high-quality shrimp larvae. In larviculture, quality feeds containing a high content of highly unsaturated fatty acids (HUFA) and ingredients that stimulate stress and disease resistance are essential to produce healthy shrimp larvae. In the present study, Penaeus monodon postlarvae (PL15) were fed for 25 days on an unenriched Artemia diet (control; A) or on a diet of Artemia enriched with either HUFA-rich liver oil of the trash fish Odonus niger (B), probionts [Lactobacillus acidophilus (C1) or yeast-Saccharomyces cerevisiae (C2)] or biomedicinal herbal products (D) that have anti-stress, growth-promoting and anti-microbial characteristics. P. monodon postlarvae fed unenriched Artemia exhibited the lowest weight gain (227.9 ± 8.30 mg) and specific growth rate (9.95 ± 0.05%), while those fed the HUFA-enriched Artemia (B) exhibited the highest weight gain and specific growth rate (362.34 ± 12.56 mg and 11.77 ± 0.08%, respectively). At the end of the 25-day rearing experiment, the shrimp postlarvae (PL40) were subjected to a salinity stress study. At both low and high (0 and 50‰) salinities, the group fed the control diet (A) experienced the highest cumulative mortality indices (CMI) 935.7 ± 2.1 and 1270.7 ± 3.1, respectively. Those fed diet D showed the lowest stress-induced mortality, and CMI were reduced by 31.1 and 32.3% under conditions of low and high salinity stress, respectively. A 10-day disease challenge test was conducted with the P. monodon postlarvae (PL40–PL50) by inoculating the shrimp with the pathogen Vibrio harveyi at the rate of 10⁵–10⁷ CFU/ml in all rearing tanks. P. monodon postlarvae fed probiont-encapsulated Artemia diets (C1 and C2) exhibited the highest survival (94.3 and 82.3%, respectively) and lowest pathogen load (V. harveyi) in hepatopancreas (5.2 × 10² ± 9.0 × 10 and 4.6 × 10² ± 9.0 × 10 CFU g⁻¹, respectively) and muscle (2.0 × 10² ± 6 × 10 and 1.7 × 10² ± 8.6 × 10 CFU g⁻¹, respectively) tissues. The shrimp that were fed the unenriched Artemia (Control; A) showed the lowest survival (26.33%) and highest bacterial load in the hepatopancreas (1.0 × 10⁵ ± 5 × 10³ CFU g⁻¹) and muscle (3.6 × 10⁴ ± 6 × 10² CFU g⁻¹). The shrimp fed the herbal product (D)-enriched Artemia also exhibited enhanced survival and reduced V. harveyi load in the tissues tested compared to the control diet (A) group. The results are discussed in terms of developing a quality larval feed to produce healthy shrimp larvae.     

Immunostimulating influence of herbal biomedicines on nonspecific immunity in Grouper Epinephelus tauvina juvenile against Vibrio harveyi infection

Herbals such as Cynodon dactylon, Piper longum, Phyllanthus niruri, Tridax procumbens, and Zingiber officinalis were extracted with acetone, benzene, butanol, and petroleum ether and screened against the pathogen Vibrio harveyi isolated from the infected Grouper Epinephalus tauvina. Among the different solvent extractions screening to V. harveyi, petroleum ether extracts were suppressed significantly (P < 0.05). Equal proportions of the all-plant extracts were mixed with the artificial feeds at concentrations of 100, 200, 400, and 800 mg kg⁻¹ of diet and fed to grouper juveniles of 20 ± 2 g average weight for a period of 60 days. Every 20 days, fish juveniles were challenged with V. harveyi and the immune response was studied. The herbal diets significantly (P < 0.05) increased the survival, growth, and immune responses compared to the control group. The herbal diets were significantly improved (P < 0.01) in immune parameters such as phagocytic activity and albumin–globulin (A–G) ratio. Among the different concentrations of the herbals in the diet, the 400 mg kg⁻¹ diet was the most effective in the experiment.     

Isolation and partial characterization of bacteriophages infecting Vibrio harveyi from shrimp farm effluent water

Bacteriophage isolated from the semi-intensive culture of Pacific white leg shrimp Litopenaeus vannamei infects the luminous bacteria Vibrio harveyi. Lytic activity and lytic spectrum results revealed that the isolated phage had strong lytic activity in V. harveyi, V. parahaemolyticus, and V. vulnificus. Biofilm inhibition activity was performed against different pathogenic vibrios on high-density polyethylene (HDPE) template and the result revealed that the phage effectively inhibited the biofilm formation in V. harveyi. Spectrophotometric assay performed for lytic activity of the isolated phage in V. harveyi liquid culture showed that the phage significantly decreased the V. harveyi cell densities at different time intervals (P?<?0.05). To study the stability of phage at different temperature and pH revealed that the phage withstands the temperature ranged between 40 and 70 °C and the pH of 4 and 9 at a significant level (P?<?0.001). One-step growth curve depicted that the burst size gradually increased to a significant level and reached the maximum of 90% at 180 min (P?<?0.05). This study concluded that the isolated phage had specific activity against pathogenic V. harveyi infections.

     

Influence of the antibacterial herbs, Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia on the survival, growth and bacterial load of Penaeus monodon post larvae

The indiscriminate use of antibiotics and chemicals in shrimp hatcheries has led to biomagnification and that in turn could lead to rejection of a whole consignment. The application of the bioencapsulation technique as a tool for curative treatment in shrimp larvae was investigated. Herbs having antibacterial properties such as Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia (methanolic extracts) were bioencapsulated in Artemia and fed to Penaeus monodon post larvae PL 1–25. The post larvae were reared in a medium inoculated with pathogenic bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Vibrio sp. Post larvae reared in the non-inoculated water and fed with non-enriched Artemia exhibited 90% survival, highest specific growth rate (12.43%) and reduced bacterial load. P. monodon reared in the bacterial inoculated water and fed with the non-enriched Artemia exhibited the lowest survival (10–30%), specific growth rate (8.42–9.1%) and increased bacterial load (2.86 × 10³ to 3.76 × 10⁵ cfu/g). The methanolic extracts of the herbs helped to increase survival and specific growth rate and reduced bacterial load in the P. monodon culture system. Among the three herbal extracts, P. corylifolia enriched Artemia fed post larvae showed the tendency to higher survival (>50%), growth rate (11.5 averaged) and low bacterial load (1.12 × 10⁵ cfu/g). This revised version was published online in August 2006 with corrections to the Cover Date.     

Herbal biomedicines: a new opportunity for aquaculture industry

Hormones, antibiotics, vitamins and several other chemicals have been tested in aquaculture operations for various remedies. Even though they give positive effects, they cannot be recommended due to their residual and other side effects. The alternative herbal bio-medicinal products in the aquacultural operations, that have the characteristics of growth promoting ability and tonic to improve the immune system, act as appetite stimulators. They increase consumption, induce maturation, and have antimicrobial capability and also antistress characteristics that will be of immense use in the culture of shrimps and other fin fishes without any environmental and hazardous problems. Herbal compounds such as phenolics, polyphenols, alkaloids, quinones, terpenoids, lectines and polypeptides have been shown to be very effective alternatives to antibiotics and other synthetic compounds. The present paper is presented after a careful review of more than 50 herbal plants for their biological effects such as growth promotion, immunostimulation, antistress, antibacterial, antifungal, antivirals, appetite stimulators and aphrodisiac.     

Effect of dietary soybean lecithin on growth parameters,digestive enzyme activity,antioxidative status and mucosal immune responses of common carp (Cyprinus carpio)

Common carp, Cyprinus carpio (13.54 ± 0.48 g), were fed soybean lecithin‐incorporated diets for 8 weeks. After the feeding trials, the different groups of fishes were assessed by survival, growth parameters, digestive enzymes, antioxidant enzyme activities, mucosal immune parameters, etc. The lecithin‐incorporated diets (10–30 g kg^–1) helped to increase the weight gain significantly (p < .05) compared with control groups; activity of digestive enzymes was also significantly (p < .05) higher compared with experimental groups. The antioxidant enzymes including catalase and serum superoxide dismutase were significantly (p < .05) higher in the C. carpio fed lecithin‐incorporated feed than the ones fed the control feed. The serum thiobarbituric acid‐reactive substances and serum glutathione peroxidase activity did not differ significantly (p > .05) between control and experimental groups. The positive correlation supported that antibacterial activity was significantly (p < .05) increased with increasing concentration of lecithin in the diets. Among the different mucosal immune parameters, lysozyme, protease and esterase activities were significantly (p < .05) increased with lecithin‐incorporated diets. The results of this study confirm that 20 and 30 g kg^–1 lecithin inclusion in diets helped to enhance the activities of digestive enzymes, antioxidant enzymes, antibacterial factors and mucosal immune parameters in C. carpio.     

A study on the influence of selected Indian herbal active principles on enhancing the immune system in Fenneropenaeus indicus against Vibrio harveyi infection

Five herbs such as Acalypha indica, Hygrophila spinosa, Picrorhiza kurooa, Tinospora cordifolia and Zingiber officinale were selected to screen for the in vitro immunostimulant activity against the shrimp pathogen Vibrio harveyi using different organic polar and non-polar solvents. After initial screening, the best extract of the five herbs was mixed thoroughly in equal proportion and made it an artificial immunostimulant diets (ISD) such as ISD-1(500), ISD-2 (1,000), ISD-3 (1,500) and ISD-4 (2,000) mg?kg^?1. The control diet was prepared without adding immunostimulant herbal extracts. The Indian white shrimp, Fenneropenaeus indicus having the weight of 8.0?±?1.0?g, were fed with the above diets for 60?days. After the completion of feeding trail, they were challenged with V. harveyi at the rate of 10⁷CFU/ml and found out the cumulative mortality, growth parameters, haematological and immunological parameters. The weight gain and specific growth rate were significantly (P?<?0.05) increased from the control group to experimental group. The immunostimulant active principles helped to decrease the coagulation time and improved the total haemocyte count (THC), phagocytosis, phenol oxidase (PO) activity, haemagglutinin activity and bacterial clearance.