Study of Bacillus licheniformis Fermented Products on Swine Dysentery and Broilers Necrotic Enteritis Prevention

• Main Authors: Horng Yi-Bing, 洪亦秉
• Other Authors: Cheng Yeong-Hsiang
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/tc2tz5

碩士 === 國立宜蘭大學 === 生物技術與動物科學系動物科學碩士班 === 106 === Abstract This study investigated the effectiveness of antibacterial substances produced by solid fermentation of Bacillus licheniformis against Brachyspira hyodysenteriae and Clostridium perfringens. Three experiments were designed as follows: Test 1. To optimize the surfactin production by fermentation of Bacillus licheniformis. Molasses were used as carbon sources for reduction of fermentation costs. The results showed that the 5 grams of molasses can increase the production of surface by Bacillus licheniformis. Increasing the nitrogen source from the 25 grams to 50 grams also increased the production of surfactin. Test 2, To study the effect of surface elements on Helicobacter erythropolis and Clostridium perfringens. The test use the minimum inhibitory concentration, inhibition zone, conjugated focus microscope and electron microscope to observe the effect of surface. The change in the surface tension of the water can be used as a basis for inferring the concentration of surface elements. A 10 ppm surface element can inhibit the Helicobacter pylori, and the survival of 500 ppm Helicobacter pallidum is reduced to 23%. For Clostridium perfringens, 25 ppm has an inhibitory effect, and 500 ppm can kill 63% of Clostridium perfringens. The use of MIC in the zone of inhibition results in 50 ppm of the Helicobacter pallidum with a 0.3 mm zone of inhibition, 500 ppm with a 1.4 mm zone of inhibition; Clostridium perfringens has a zone of 0.3 mm at 7.8 ppm, 250 ppm has a 0.8 mm zone of inhibition. Confocal microscopy observation, using SYBR green (live cells), PI (dead cells) and Hoechest (background values) were stained separately, and it was observed that the erythropoietin had a 36% killing effect at 50 ppm for 30 minutes. Increased to 250 ppm up to 83%, up to 46% and 94% in one hour treatment; in addition, Clostridium perfringens has a 41% killing effect at 25 ppm and 48 ppm at 250 ppm. The treatment time was extended to one hour and increased to 46% and 84%, respectively. Finally, the scanning electron microscope (SEM) was used to observe the erythropoiesis. The results showed that the surface of the Helicobacter pallidum was damaged at 10 ppm. Under the treatment of 250 ppm, the cells could be completely broken, forming a broken and broken type. Maintaining its original spiral shape, this result is consistent in the half-hour and one-hour treatment groups, and it can be known that surface substances have the effect of inhibiting or killing both pathogenic bacteria. Test 3: A white broiler animal test was carried out using a fermentation product of Bacillus, and the effect on the growth performance of the chicken and the necrotic enteritis caused by Clostridium perfringens was tested. 1% and 2% fermented materials were added to the diet, and the number of spores of B. licheniformis was 1.2 × 107 CFU/g and 2.4 × 107 CFU/g. The test treatment group was the control group (no addition) and the positive control group (Add enromycin 0.5 kg / ton, 500 ppm), treatment group A (fermentation 1 kg / ton) and treatment group B (fermentation 2 kg / ton), each group of three replicates, each cage five total 60 broilers, the test schedule was 35 days, and 1 mL of Clostridium perfringens liquid was administered orally on the 17th, 18th and 19th days of feeding, and the dose was 1 x 108 CFU/mL. The results showed that the growth performance, intestinal flora and inflammatory index of the feed-added Bacillus licheniformis fermented group were better than the control group.