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Studies on Effect of Multispecies Probiotics on Nonspecific Immunity and Resistance to WSSV of Litopenaeus Vannamei

Author: LiGuiYing
Tutor: SongXiaoLing;MaiKangSen
School: Ocean University of China
Course: Hydrobiology
Keywords: Probiotics Immune enzyme activities Digestive enzyme activities Immune-related genes Anti - WSSV infectivity
CLC: S968.22
Type: Master's thesis
Year: 2011
Downloads: 117
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High density intensive shrimp farming industry worldwide rise by the prawn itself and the external environment caused a variety of bacterial, viral diseases severely restricted the sustained and healthy development of the industry, but it also brings serious economic losses. Various measures in this context came into being, but traditional methods of treatment of antibiotics, disinfectants and pesticides has been questioned constantly being explored alternative method to improve the body's own immune system and resistance to disease is the solution one of the important ideas of this problem. In recent years, probiotics as an alternative way to control aquaculture diseases caused by hot, detailed research reports from the good effects of probiotic applications, non-polluting and security. In this thesis, isolated several strains of probiotic bacteria from the intestinal tract of healthy shrimp add compatibility in the shrimp base feed in order to observe the nonspecific immunity and anti-virus capabilities, at the same time a part of the molecular biology analysis in order The mechanism of action of probiotics provide a theoretical basis. The main research content and results of the following aspects: 1, Bacillus firmus (Bacillus firmus), the mermaid Photobacteriosis (Photobacterium damselae) and Vibrio alginolyticus (Vibrio alginolyticus) compatibility Litopenaeus vannamei non-specific immune enzyme activities and Disease Resistance: Bacillus firmus viable cells (less than 1.0 × 10 8 CFU / g feed) and the mermaid Photobacteriosis (1%) and alginolyticus the vibrio (1%) off The live bacteria compatibility of basal diet as immunization feed. Basal diet control group throughout the experiment to take interval feeding (before validation for better feeding). Immune 15d the WSSV disease material (infectious dose 0.9g/10 tail) artificial feeding infected the infected time for 15d. During the experiment 5d shrimp hemolymph collected in each experimental group treated for superoxide dismutase (SOD), alkaline phosphatase (AKP), acid phosphatase (ACP), nitric oxide synthase (NOS) and lysozyme enzyme (UL) Determination of vitality. Infection prawn deaths recorded daily and cumulative mortality statistics of the control group and the experimental group. The results show that each immunohistochemistry superoxide dismutase (SOD), alkaline phosphatase (AKP), acid phosphatase enzyme (ACP), nitric oxide synthase (NOS) activity overall was significantly higher than that of the control group, but the feed The probiotic significantly lysozyme actives. The cumulative mortality results showed: Add the Bacillus firmus mermaid Photobacteriosis the strongest antiviral potency of the mixed group (10.71%), significantly higher than the control group (64.28%), followed by Bacillus firmus and the Vibrio alginolyticus mixed group. The results confirmed: add probiotics in feed can significantly increase the Litopenaeus vannamei non-specific immune activity of viral resistance, which especially the complex bacteria Bacillus firmus mermaid luminous Bacillus group effect. 2, strong Bacillus (Bacillus firmus), the mermaid Photobacteriosis (Photobacterium damselae) and alginolyticus Vibrio (Vibrio alginolyticus) compatibility Litopenaeus vannamei intestinal digestive enzyme activity and intestinal flora: according to the first part of The compatibility of the preparation of immune feed, experimental grouping and processing the same. Take shrimp intestinal homogenate per 5d under sterile conditions during the experiment, a part used to measure the activity of digestive enzymes (pepsin, amylase and lipase), part of the gradient dilution coating 2216E solid medium for bacterial colony counting. The results showed that probiotic feed immunohistochemistry of pepsin, amylase and lipase showed a certain regularity, the first rise in the overall immune stage and infection stage and then decreased, and the maximum value of the experimental stage generally appear in the experimental 25d generally fell back to the beginning of the experiment level again at the end of the experiment. Digestive enzyme activities during the entire experiment immunohistochemistry most stages were significantly higher than those in the control group (P lt; 0.05). This law change has a regulatory role of probiotics on intestinal Litopenaeus vannamei diets, probiotics such regulation is not unlimited increase drops can be seen by the immune during the first rise. Trend increased and then decreased in the stage of infection, probiotic able to make the appropriate response to viral infection, but its reaction is not constant. Changes in the intestinal flora the of Bacillus firmus mermaid Photobacteriosis mixed bacteria group can cultivate the total number of bacteria and the total number of dominant bacteria was significantly higher (P lt; 0.05), the rest of immunohistochemistry difference was not significant (P gt; 0.05 ). These results suggest that the diet containing probiotics can significantly improve shrimp some regulation of intestinal digestive enzyme activity and intestinal flora with probiotics play a role in the host body. Three strong Bacillus (Bacillus firmus), mermaid light bacilli (Photobacterium damselae) and Vibrio alginolyticus (Vibrio alginolyticus) different compatibility Litopenaeus vannamei immune-related gene expression: experimental basal diet as a control group, not less than 1.0 × 10 8 to add Bacillus firmus viable cells (CFU / g feed) mermaid luminous bacillus (1%) and Vibrio alginolyticus (%) feed for immunohistochemistry. Groups of three repeat experiments during interval feeding. Each 5d take each experimental group 10-15 tail shrimp the extracted hemolymph centrifugation after collection of blood cells for RNA extraction, β-actin as reference gene, Litopenaeus vannamei STAT gene, superoxide dismutase (SOD) gene and prophenoloxidase (proPO) gene with the Rotor-gene 3000 real-time quantitative PCR, 2 -ΔΔCT Law relative expression level of each gene mRNA analysis. The results show that in the feed probiotic prophenoloxidase (proPO) gene relative expression but not significantly higher than the control group, STAT and superoxide dismutase (SOD) gene expression relative little impact. 4,4 shrimp environmental strains of symbiotic bacteria research on of Litopenaeus vannamei prebiotic effect: the study was designed to explore isolated from healthy where several strains of bacteria of the digestive tract and breeding of shrimp water: Exiguobacterium (Microbacterium spp), thuringiensis Bacillus ( Bacillus thuringiensis), alginolyticus (Vibrio alginolyticus) and Bacillus cereus (Bacillus cereus) Litopenaeus vannamei prebiotic effect. Litopenaeus vannamei formulated immune feed the strains of single or composite basal diet, basal diet control group, groups of three repeat individual quality (6.1 ± 0.2) g for a period of 30d of breeding experiments. 5d each sampling, serum alkaline phosphatase (AKP), the activity of nitric oxide synthase (NOS), superoxide dismutase (SOD), lysozyme (UL) and phenol oxidase (PO) immune indicators explore the the shrimp environment symbiotic bacteria as feed additives Litopenaeus vannamei level of non-specific immune; feeding after immunization feed 16d tail 1.00g/10 dose directly fed infected with white spot syndrome virus ( WSSV) of shrimp disease expected to calculate cumulative mortality rate of each experimental group to analyze the impact of the shrimp environment symbiotic bacteria as feed additives Litopenaeus vannamei anti-virus ability. Experimental results show that immunohistochemistry Where: the shrimp serum add bacteria immunohistochemistry SOD, PO, AKP, UL and NOS activity were not significantly higher, and immunohistochemistry immune related activity than the control group; Prawns anti WSSV infection ability improved significantly. These results show that: the experimental diets 4 bacteria and failed to improve the of Litopenaeus vannamei nonspecific immunity levels and ability to resist disease, temporarily as shrimp immunostimulants candidate strains.

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CLC: > Agricultural Sciences > Aquaculture, fisheries > Aquaculture technology > A variety of seafood, animal and plant breeding > Crustacean culture > Shrimp farming
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