Application Of Bacillus Spp. And Traditional Chinese Medicine In The Feed Of Shrimp Litopenaeus Vannamei

A series of experiments were conducted to investigate the application of diatary Bacillus spp. and traditional Chinese medicine on shrimp Litopenaeus vannamei, based on microbiological, physiological, nutritional, immunological and ecological methods. The results are summarized as followed:1. A trial was conducted to investigate the effect of traditional Chinese medicines (TCM) on growth of Bacillus licheniformis. The growth of B. licheniformis could be promoted in some concentrations of TCM. Therefore, they were potential to be used together in the feed.2. The basal diet (C); with 0.20% TCM (M), with 0.30% Bacillus spp. and 0.20% TCM (BM1), with 0.15% Bacillus spp. and 0.10% TCM (BM2), with 0.30% Bacillus spp. (B) was used to fed white shrimp Litopenaeus vannamei (1.91±0.03 g) in order to assess the survival, growth, body composition and digestive enzyme activity. Growth measured as weight gain was significantly (P<0.05) higher in shrimp fed with BM2, BM1 and M compared to that of C. However no significant differences were found among B, BM1 and M. In the case of specific growth rate, the shrimp fed with BM1 and BM2 exhibited significantly (P<0.05) higher values than that of the C. The contents of body moisture, crude protein and ash seemed to be unaffected by the feed supplements, though lipid content was found to be significantly (P<0.05) different among the treatments. The shrimp fed with BM1 and BM2 had the lowest and highest lipid contents respectively. The digestive enzyme activity assessed using shrimp hepatopancreas revealed that the activities of protease and amylase in shrimp fed with BM2 were significantly (P<0.05) higher than those of the C at the end of 2nd and 6th week. However, better performance of the specific amylase activity was shown by the shrimp fed with B at the end of 8th week.3. A feeding experiment was conducted to investigate the effects of Bacillus and traditional Chinese medicines (TCM) on growth and intestinal bacterial flora of shrimp L. vannamei. Four experimental diets were formulated by adding the Bacillus and TCM to basal diet (C): 0.2% TCM (M), 0.20% TCM+0.30% Bacillus (BM1), 0.10% TCM+0.15% Bacillus (BM2), 0.30% Bacillus (B). The concentration of Bacillus was 109 CFU/g. The TCM consisted of a mixture of herbs and plant materials Isatis tinctoria L., Isatis indigodica Fort, Forsythia suspersa Vahl, Corydalis bungeana Turez, Pogostemon cablin (blanco) Benth and Astragalus menbranaceus (Fisch.) Bge. Acclimatized shrimp with an average initial weight 1.91±0.03 g were selected and stocked into 15 cement tanks at a stocking density of 80 shrimp. Each aquarium contained 1500-l of sand filtered seawater with a flow rate of 0.9-l min-1. The water was continuously aerated with two air stones. Each diet was given to shrimp four times (07:00, 12:00, 17:00, 23:00) a day to saturation for a period of 56 days. During the experimental period, the water temperature fluctuated between 19.6 and 25.2℃, salinity between 30 and 31. At the end of feeding trial, survival ranged from 95.83% to 98.33% with no significant difference (P>0.05) among all groups. Growth measured as final weight was slightly higher (P>0.05) in shrimp fed with BM1, M and B than that of C, and there was a significant difference (P<0.05) between BM2 and C. In case of weight gain (WG) and specific growth rate (SGR), the shrimp fed with M, BM1 and BM2 exhibited significantly (P<0.05) higher values than that of the C and B. Of the five diets, BM2 exhibited the best performance followed by BM1. The combination of Bacillus spp. and TCM, therefore, proved to be more effective and economically viable. The total bacteria counts and Vibrio counts in shrimp fed with M was the most, which fed with BM1 and BM2 was the least. In each group 30 bacterial strains were purified and identified. These bacterial strains belonged to 12, 8, 6, 5, 7 genera in C, M, BM1, BM2 and B respectively. Most of the bacteria were gram negative bacteria, they counted a percentage of 86% to 93% in each group. Vibrio spp., Photobacterium spp. and Pseudomonas spp were the predominant bacterial flora, however, their values varied in different groups. The results showed that the diets supplemented with TCM and Bacillus also changed the number and composition of intestinal bacterial flora.4. A feeding experiment was conducted to investigate the effects of Bacillus and traditional Chinese medicines (TCM) on the survival, growth, body composition, digestive enzyme activity and serum biochemical values of shrimp L. vannamei. Six experimental diets were formulated by adding the Bacillus and TCM to basal diet (Diet0, Control): 0.10% TCM+0.10% Bacillus (Diet11), 0.10% TCM+0.20% Bacillus (Diet12), 0.10% TCM +0.30% Bacillus (Diet13), 0.20% TCM+0.10% Bacillus (Diet21), 0.20% TCM+0.20% Bacillus (Diet22), 0.20% TCM +0.30% Bacillus (Diet23). After 56 days of feeding, survival ranged from 76.19% to 84.76% with no significant difference (P>0.05) among all groups. Growth measured as final weight, specific growth rate (SGR), protein efficiency ratio (PER), protein productive value (PPV) and feed efficiency ratio (FER) were improved as the addition of TCM and Bacillus increasing. In the case of final weight and SGR, the shrimp fed with Diet23 exhibited significantly (P<0.05) higher values than that of Diet0. The contents of body moisture, crude protein and ash seemed to be unaffected by the feed supplements, though lipid content was found to be significantly (P<0.05) different among the treatments. The shrimp fed with Diet23 had a higher lipid contents than which fed with Diet0, Diet13 and Diet22 (P<0.05). The digestive enzyme activity assessed using shrimp hepatopancreas and intestines revealed that the activities of protease and amylase in shrimp were significantly (P<0.05) enhanced by the feed supplements, but amylase of hepatopancreas was except. The alkaline phosphatease and acid phosphatease values were significantly affected by the interaction of Bacillus and TCM (P<0.05). Serum protein and glucose content were enhanced as the addition of TCM and Bacillus increasing. However, Serum triglyceride in shrimp of low TCM groups (Diet11, Diet12 and Diet13) and cholesterol in shrimp of high Bacillus groups (Diet12, Diet13 and Diet23) were significantly higher than that of Control (Diet0).5. A feeding experiment was conducted to investigate the effects of Bacillus and traditional Chinese medicines (TCM) on immunity of shrimp L. vannamei. Six experimental diets were formulated by adding the Bacillus and TCM to basal diet (Diet0, Control): 0.10% TCM+0.10% Bacillus (Diet11), 0.10% TCM+0.20% Bacillus (Diet12), 0.10% TCM +0.30% Bacillus (Diet13), 0.20% TCM+0.10% Bacillus (Diet21), 0.20% TCM+0.20% Bacillus (Diet22), 0.20% TCM +0.30% Bacillus (Diet23). After 56 days of feeding, the total haemocyte count (THC), phenoloxidase (PO), lysozyme activity, superoxide dismutase (SOD) and total antioxidant was measured. The data indicated that THC was significantly affected by Bacillus so that it was higher in shrimp fed with Diet 12, Diet13 and Diet23 than that of others (P<0.05). The value of PO was improved by high content of TCM. The lysozyme activity of shrimp in most experimental diet groups was higher than that of control. However, SOD seemed to be unaffected by the feed supplements. In the case of total antioxidant, the shrimp fed with Diet23 exhibited significantly (P<0.05) higher values than that of other groups except for Diet21.6. A feeding experiment was conducted to investigate the effects of Bacillus and traditional Chinese medicines (TCM) on the survival, growth, body composition, digestive enzyme activity and serum biochemical values of shrimp L. vannamei. Two practical diets were used as basal diets in the experiment, one of which is high quality diet contained 29% fish meal (basal diet 1), the other one of which is low quality diet contained 17% fish meal (basal diet 2). Six experimental diets were formulated by adding the Bacillus and TCM to the basal diets: Basal diet 1 (Diet10), Basal diet 1+0.10% TCM+0.15% Bacillus (Diet11), Basal diet 1+0.20% TCM+0.30% Bacillus (Diet12), Basal diet 2 (Diet20), Basal diet 2+0.10% TCM + 0.15% Bacillus (Diet21), Basal diet 2+0.20% TCM+0.30% Bacillus (Diet22). After 56 days of feeding, shrimp in the groups fed with basal diet 2 had a higher survival than that of basal diet 1. Growth measured as final weight, specific growth rate (SGR), protein efficiency ratio (PER), protein productive value (PPV) and feed efficiency ratio (FER) were significantly affected by the different basal diets and supplements. In the case of final weight and SGR, the shrimp in the groups fed with basal diet 1 exhibited significantly (P<0.05) higher values than that of basal diet 2. While for the same basal diet group, PER, PPV and FER of shrimp were improved by the supplement. The contents of body compositions were also affected by the feed supplements and different basal diets. The digestive enzyme activities assessed using shrimp hepatopancreas and intestines revealed that the activities of protease and amylase in shrimp were significantly (P<0.05) enhanced by the feed supplements. The alkaline phosphatease in shrimp hepatopancreas were significantly (P<0.05) enhanced by the feed supplements too. The alkaline phosphatease and acid phosphatease values were significantly higher in shrimp fed with basal diet 2 than basal diet 1. Serum cholesterol and glucose contents had a significant difference among different basal diet groups. However, Serum protein and glucose contents were enhanced by the supplements.7. A feeding experiment was conducted to investigate the effects of Bacillus and traditional Chinese medicines (TCM) on immunity of shrimp L. vannamei. Two practical diets were used as basal diets in the experiment, one of which is high quality diet contained 29% fish meal (basal diet 1), the other one of which is low quality diet contained 17% fish meal (basal diet 2). Six experimental diets were formulated by adding the Bacillus and TCM to basal diets: Basal diet 1 (Diet10), Basal diet 1+0.10% TCM+0.15% Bacillus (Diet11), Basal diet 1+0.20% TCM+0.30% Bacillus (Diet12), Basal diet 2 (Diet20), Basal diet 2+0.10% TCM + 0.15% Bacillus (Diet21), Basal diet 2+0.20% TCM + 0.30% Bacillus (Diet22). After 56 days of feeding, the total haemocyte count (THC), phenoloxidase (PO), lysozyme activity, superoxide dismutase (SOD) and total antioxidant were measured. The data indicated that THC and lysozyme activity were significantly affected by the feed and supplement, while total antioxidant was significantly (P<0.05) affected by the supplement and their interaction. As a whole, THC, PO, lysozyme activity, SOD and total antioxidant were enhanced more or less by adding the supplement in different diets. The values of THC and lysozyme activity in basal diet1 groups were significantly enhanced by the supplement. The values of THC and total antioxidant in basal diet2 groups were significantly enhanced by the supplement too. Total antioxidant were significantly (P<0.05) higher in Diet20 than in Diet10, which indicated that the immunity were also affected by the feed.8. A feeding experiment was conducted to investigate the effects of Bacillus and traditional Chinese medicines (TCM) on the growth, intake, faecal loss, non-faecal loss, body composition and apparent digestibility coefficients (ADCs) of shrimp L. vannamei. Two practical diets were used as basal diets in the experiment, one of which is high quality diet contained 29% fish meal (basal diet 1), the other one of which is low quality diet contained 17% fish meal (basal diet 2). Six diets were formulated by adding the Bacillus and TCM to the basal diets: Basal diet 1 (Diet10), Basal diet 1+0.10% TCM+0.15% Bacillus (Diet11), Basal diet 1+0.20% TCM+0.30% Bacillus (Diet12), Basal diet 2 (Diet20), Basal diet 2+0.10% TCM + 0.15% Bacillus (Diet21), Basal diet 2+0.20% TCM+0.30% Bacillus (Diet22). After 56 days of feeding, the results showed that growth, intake, feed conversion were increased while faecal loss and loss in unit weight gain were decreased by adding the supplement. The element of carbon, nitrogen and phosphorus in growth and intake were improved while which in faecal loss was reduced by adding the supplement. The value of intake, final weight, weight gain and feed conversion in shrimp of basal diet1 groups were higher than that of basal diet2 groups, while the value of loss in unit weight gain were reversed. Body carbon and nitrogen seemed not to be affected by the feed and supplement, while body phosphorus, faecal carbon, faecal nitrogen and faecal phosphorus were affected by them. ADCs of dry matter, carbon, nitrogen and phosphorus were improved by the supplement and also affected by the feed.