| Summary: | 碩士 === 國立臺灣海洋大學 === 水產養殖學系 === 99 === Probiotics, which are micro-organisms and their products with health benefit to the host, have used in aquaculture as a means of disease control, supplementing or even in some cases replacing the use of antimicrobial compounds. Water quality is the most important essentials in aquacluture industry. In past, it has been shown most probiotics can for improving water quality, are phototrophic bacteria and nitrifying bacteria.
The objective of present study is to stablish quantitation method to measure specific bacteria in complex microorganisms and applications of Complex Beneficial Microorganisms (CBM) in water quality improvement and immunity enhancement in white shrimp（Litopenaeus vannamei）. To accomplish this objective, several specific aims were established ; 1）apply real-time quantitative polymerase chain reaction
(Real-Time Quantitative PCR, Q-PCR) to quantitate composite probiotic of CBM which contaning various of strains of microorganisms, 2）apply of CBM in simulated wastewater treatment for improving water quality, 3）use CBM as a direct-fed-microbial to enhance innate immunity and
disease resistance in white shrimp. In present study, Q-PCR quantification of CBM bacteria, R. palustris, R. rubrum, R. sphaeroides,L. pentosus, L. lactis, Bacillus, S. fibuligera, O. anthropi, T. pantotropha, A. denitrificans, P. citrinum, C. globosum, A. oryzae, M. verrucaria, A.niger in symbiotic amplification culture process were; 4.10 × 10 9 , 2.56 × 10 9 , 1.39× 10 8 , 3.57 × 10 8 , 7.90 × 10 8 , 1.42 × 10 8 , 2.13 × 10 8 , 2.75 × 10 8 ,4.69 × 10 8 , 5.22 × 10 8 , 3.85 × 10 8 CFU‧mL -1 respectively; the cells number in first molasses amplification culture process were; 1.58 × 10 7~ 7.94 × 10 8 , 3.027 × 10 6 ~ 3.92 × 10 8 , 7.6 × 10 6 ~ 6.4 × 10 8 , 9.91 × 10 6 ~ 5. 16 × 10 8 , 9.91 × 10 6 ~ 6.77 × 10 8 , 3.55 × 10 6 ~ 7.36 × 10, 4.89 × 10 6 ~ 4.23 × 10 8 , 8.9 × 10 6 ~ 1.40 × 10 8 , 5.57 × 10 6 ~ 11.40 × 10 8 , 1.16 × 10 6~ 3.04 × 10 8 , 1.6 × 10 6 ~ 5.08 × 10 8 CFU‧mL-1 respectively; the cell numbers in second molasses amplification culture process were; 0.58 × 10 7 ~ 7.94 × 10 8 , 3.02 × 10 6 ~ 3.92 × 10 8 , 1.6 × 10 6 ~ 2.4 × 10 7 , 3.12 × 10 6 ~ 1.41 × 10 7 , 3.12 × 10 5 ~ 1.41 × 10 7 , 1.84 × 10 6 ~ 7.5 × 10 7 , 1.30 × 10 6 ~ 4.17 × 10 7 , 4.67 × 10 6 ~ 2.76 × 10 7 , 6.1× 10 5 ~ 2.9 × 10 7 , 6.1× 10 5 ~ 3.75 × 10 7 , 1.04× 10 6 ~ 2.29 × 10 7 CFU‧mL -1 respectively. In water quantity treatment tests, adding 100 ppm into freshwater aquaculture wastewater, marine wastewater, and industrial wastewater, had shown that the ammonia and nitrite levels reducing significantly. Extra carbon source additation can enhance denitrification process and reducing ammonia and nitrite level significantly. Using CBM as DFM experiment in white shrimp, the result indicated that the expression of haemocyanin,
crustin and penaeidin genes increased, the mRNA levels of haemocyanin, crustin were twice amount than the control group. After CBM administration, the results of pathogens challenged test demonstrated that
shrimp fed with survival rate CBM can be improved about 30% after Vibrio harveyi infection. The group fed with CBM had 20% higher survival rate after WSSV infection.
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