Reserch On The Microhe On Inteinal Flora Of Apostichous Ijapncus On Prevtion Of Pathogeniclity

Sea cucumber belongs to Echinodermata and has a history of six hundreds million years. Sea Cucumber is on the top of the "eight treasures" list because of the value of food and medicinal. Apostichpus japonicus has the highest nutritional value among the Sea Cucumbers. The Apostichpus japonicus produced in Liaoning and Shandong is the important species in China. Due to the nutritional value and significant economic benefits, Apostichpus japonicus breeding industry had increased around the Pohai sea in recent years, and has become the pillar industry after the Sea tangle, Shrimp, Scallops, marine fish. However, due to the tachy-development and non-standard operations, Apostichpus japonicus generate a large number of pathogenic bacteria,which resulted leading to the lower survival rat and great economic loss. On the disease control in aquaculture, antibiotics are still used as the primary means. But with a large number of antibiotic medication, the side effects are becoming increasingly more serious, food safety problem has be noticed to full attention. Thus, it is significant to the Apostichpus japonicus industry to develop non-toxic, no residue, no pollution new Chinese herbal medicine, in this way, it can enhence immunity of the sea cucumber, prevent and treat Apostichpus japonicus disease in order to promote the growth of sea cucumber.Apostichpus japonicus " skin ulceration disease " has become the research priorities of sea cucumber breeding because of its contagious, high mortality. In this paper, the cause of the " skin ulceration disease " was explored based on the separation and identification of the intestinal flora in healthy and diseased sea cucumber. Flora in the intestinal of healthy and diseased sea cucumber and focus on infection were separated and identified, respectively. The results showed that bacteria in the intestine of sea cucumber was diverse, and the 15 bacteria had been identified through 16SrDNA, and the bacillus was the most important, combined with morphological, physiological and biochemical, the dominant bacteria LNUB417 was identified as Bacillus subtilis, LNUB419 as Bacillus licheniformis. Intestinal microflora of Apostichpus japonicus with " skin ulceration disease " had marked change in structure and 16 bacteria were obtained, dominant LNUB415 was Bacillus cereus. A dominant LNUB416 was obtained in Apostichpus japonicus with " skin ulceration disease " and foucs on infection and was identified as planococcus. It was inferred that " skin ulceration disease "was mainly due to pathogens, alterred the normal flora balance, resulting in the decrease in sea cucumber resistance.Astragalus, Forsythia, Folium, Houttuynia, Radix, Berberine, Rhubarb, Baicalin, Chuanxiong, Honeysuckle, Licorice, Gall and the common antibiotics were used in the separation of bacteria on the antibacterial test. The results showed that berberine had obvious inhibiting effect on LNUB415 and LNUB416 and the 2 strains had sensitivity to norfloxacin, cefoperazone, nitrofurantoin. Suspend the LNUB415 and LNUB416 into bacterial liquor, the use the dilution gradient injected via- hypodermic. The results showed that LNUB415 was one of the conditions of pathogenic bacteria for the " skin ulceration disease " and the death rate increased with the concentration of bacterial suspension, the mortality rate up to 100% at the high concentration. Choose the berberine(B1) and norfloxacin(B2) to feed the Apostichpus japonicus with disease, the death rate of B1 decrease to 30% and the B2 decrease to 20%. LNUB416 can also lead to "skin ulceration disease", but the mortality did not increase as the concentration, and the death rate was 60% to 70%, after 7 days subcutaneous injection of symptoms, a longer incubation period. The group given Berberine had a markable effect, the death rate was the same as the group with norfloxacin, which was 20%. Berberine had a preventation effect on the " skin ulceration disease " of Apostichpus japonicus. So it is suggested to choose Chinese herb as the substitute of antibiotics, because it has no resistance, no residue, no pollution, etc.