Sea cucumber compound feed

[0019] Specific embodiments of the present invention will be described below.

[0020] 1. Preparation of Aspergillus niger spore suspension:

[0021] The preserved and rejuvenated Aspergillus niger fungus strain was inoculated on the PDA medium by punching, and cultured at 28°C for 3 days. After the hyphae covered the entire plate, the spores on the medium were washed off with sterile water. A suspension of Aspergillus niger spores (ie fungal seed liquid) is obtained.

[0022] Koji fermentation:

[0023] Take a certain amount of soybean meal, add water according to the ratio of material to water ratio of 80% and mix well, autoclave at 121°C for 20 minutes, and cool to room temperature. Put it into the fermentation container (sterile), insert the Aspergillus niger spore suspension (inoculation amount 10%v/w), put it in a 31℃ thermostat, and cultivate for 2 days. During the cultivation process, stir once at 24h and replenish with sterile water (replenish the lost water according to the water content and maintain the water content), and stir twice without replenishing water on the second day. After the fermentation is completed, it is dried into koji at 50°C to make the weight the same as when the soybean meal is not fermented to obtain Aspergillus niger fermented soybean meal.

[0024] 2. Preparation of Trichoderma spore suspension:

[0025] The preserved and rejuvenated Trichoderma fungal strains were inoculated on the PDA medium by punching, and cultured at 28°C for 3 days. After the hyphae spread over the entire plate, rinse off the spores on the medium with sterile water. A suspension of Trichoderma spores (ie fungal seed liquid) is obtained.

[0026] Koji fermentation:

[0027] Take a certain amount of soybean meal, add water according to the ratio of material to water ratio of 80% and mix well, autoclave at 121°C for 20 minutes, and cool to room temperature. Put it into the fermentation container (sterile), insert the Trichoderma spore suspension (inoculation amount 10% v/w), put it in a 28°C thermostat, and incubate for 2 days. During the cultivation process, stir once at 24h and replenish with sterile water (replenish the lost water according to the water content and maintain the water content), and stir twice without replenishing water on the second day. After the fermentation is completed, it is dried into koji at 50°C to make the weight consistent with the unfermented soybean meal to obtain Trichoderma fermented soybean meal.

[0028] 3. Preparation of Shewanella suspension

[0029] Inoculate the conserved sea cucumber intestinal bacteria in 2216E liquid medium, culture at 28℃, 150rp/min shaker for about 12 hours, so that the number of bacteria reaches 10 10 cfu/mL, made into bacterial suspension.

[0030] Shewanella fermentation:

[0031] After mixing a certain amount of soybean meal at a material-to-water ratio of 80%, autoclave at 121°C for 20 minutes, cool to room temperature, put it into a fermentation container, and then access the above bacterial suspension (inoculation amount 1%v/w). ), put it in a 35°C thermostat and incubate for 3 days. During the first and second days of the cultivation process, the lost water was added according to the water content and the water content was maintained. On the third day, the water was not added and stirred twice to obtain the Shewanella fermented soybean meal. After the fermentation is completed, it is dried at 50°C to make the weight of the soybean meal consistent with the unfermented soybean meal.

[0032] Feeding test

[0033] The feeding experiment was conducted on the second floor of the Engineering Center of the Technical Department of Dalian Haibao Fishery Co., Ltd. A total of four test groups were set up, each with five repetitions, using 300L tanks, each tank containing 30 sea cucumber seedlings, weighing about 400g, and feeding every afternoon. The initial daily feeding amount was 3% of the body weight. Daily observation The bait situation determines and controls the amount of bait to ensure that there is no remaining bait, and the pool is poured according to the state of the sea cucumber.

[0034] This feeding experiment started on October 10, 2015 and ended on December 14, 2015. It lasted for 64 days, and the test water temperature was 14-19°C.

[0035] The Aspergillus niger fermented soybean meal, Trichoderma fermented soybean meal, and Shewanella fermented soybean meal that have been fermented by the above three bacteria are prepared into composite fermented soybean meal at a mass ratio of 16:16:68.

[0036] The CK feed formula of the control group was: seaweed 40.3%, sea mud 45%, soybean meal 2.7%, fish meal 5%, shell powder 4.8%, yeast powder 2%, multi-dimensional 0.1%, and polysaccharide 0.1%.

[0037] The feed formula of test group A is: seaweed 40.3%, sea mud 45%, compound fermented soybean meal 2.7%, fish meal 5%, shell meal 4.8%, yeast powder 2%, multi-dimensional 0.1%, polysaccharide 0.1%.

[0038] The feed formula of test group B is: seaweed 29.5%, sea mud 45%, compound fermented soybean meal 13.5%, fish meal 5%, shell meal 4.8%, yeast powder 2%, multi-dimensional 0.1%, polysaccharide 0.1%.

[0039] The feed formula of test group C is: 16% seaweed, 45% sea mud, 27% fermented soybean meal, 5% fish meal, 4.8% shell powder, 2% yeast powder, multi-dimensional 0.1%, and polysaccharide 0.1%.

[0040] Feeding sea cucumbers with various diets, the results are as follows:

[0041] Table 1 Effects of different feed formulas on the growth of sea cucumbers

[0042]

[0043] Note: * means significant difference compared with control (P<0.05)

[0044] It can be seen from the feeding results that the feed of test groups B and C can significantly promote the growth of sea cucumbers and reduce the feed coefficient.