Lactobacillus plantarum for preventing chicken necrotizing enteritis and its application

A technology for Lactobacillus plantarum and necrotic enteritis, applied to medical preparations containing active ingredients, bacteria, microorganisms, etc.

Active Publication Date: 2019-11-05
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as the negative effects of a series of antibiotics have been reported, many countries have gradually reduced or banned the use of antibiotics as feed additives in animal production (Wade et al., 2015; Uzal FA et al., 2014; Shimizu et al. al., 2002; Myers et al., 2006), which resulted in the large-scale and epidemic occurrence of many intestinal diseases such as necrotizing enteritis (Si W et al., 2007; Nauerby et al., 2003; Peders et al., 2003)

Method used

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  • Lactobacillus plantarum for preventing chicken necrotizing enteritis and its application
  • Lactobacillus plantarum for preventing chicken necrotizing enteritis and its application
  • Lactobacillus plantarum for preventing chicken necrotizing enteritis and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1, the acquisition of Lactobacillus plantarum Lac16

[0022] 1. Prepare diluent:

[0023] Take 1 gram of soil from a vegetable field in the suburbs of Hangzhou, add it to 90 mL of sterile water, and then dilute it to 10 mL with sterile water. 0 、10 -1 、10 -2 , 10 -3 、10 -4 、10 -5 、10 -6 A total of seven concentration gradients.

[0024] 2. Cultivate:

[0025] Take 100ul of dilutions of each concentration and spread them on MRS solid medium (peptone 10g / L, yeast extract 5g / L, beef extract 10g / L, sodium acetate 5g / L, dipotassium hydrogen phosphate 2g / L, citric acid Ammonium 2g / L, magnesium sulfate 0.2g / L, manganese sulfate 0.2g / L, Tween 80 1.0mL / L, glucose 20g / L, agar 20g / L, the rest is double distilled water; the pH value is adjusted to 6.2 to 6.6, sterilized at 121°C for 20 minutes) on a flat plate, coated with a sterilized glass rod and placed upside down in a 35°C incubator for incubation.

[0026] 3. Isolation and purification of strains:

[002...

Embodiment 2

[0043] Embodiment 2, the effect of Lactobacillus plantarum Lac16 on the immune function of chicken macrophage cell line HD11

[0044] Chicken macrophage cell line HD11 according to the concentration of 5×10 5 cells / mL inoculated into 12-well cell culture plate, 1mL F12 complete medium (88% F12 basal medium + 10% chicken serum + 1% penicillin + 1% non-essential amino acid (100x)) in each well, set up control group, Lac16 group, Cp group (Clostridium perfringens) and Lac16+Cp group; specifically:

[0045] Control group: without adding inactivated lactic acid bacteria, cultured only with F12 complete medium for 3, 6, and 12 hours respectively, discarded the culture medium, washed with room temperature sterilized PBS for 3 times, added F12 complete medium and continued to cultivate for 3 hours, then harvested Sample.

[0046] Lac16 group: add 1ml containing 10 8 The cfu / mL F12 complete medium of inactivated Lactobacillus plantarum Lac16 was treated for 3h, 6h and 12h respective...

Embodiment 3

[0051] Embodiment 3, the preparation method of Lactobacillus plantarum Lac16 inoculum:

[0052] Plantarum Lactobacillus was inoculated in MRS liquid medium at a ratio of 1% (v / v), cultured at 35°C for 24 hours, centrifuged at 4°C and 5000rpm / min, the bacterial precipitate was collected, and dried at a constant temperature of 50°C for 6-8 hours. Mix it with cornstarch at 4:1 (w:w) to make dry bacteria powder, the number of viable bacteria per gram of bacteria powder is 1.0×10 8 cfu.

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Abstract

The invention discloses a plant Lactobacillus Lac16, the preserved name of which is Lactobacillus plantarum Lac16, and the preserved number is: CCTCC NO: M 2016259. It can prevent the occurrence of necrotic enteritis in chickens; inhibit the growth of Clostridium perfringens, thereby alleviating the impact of Clostridium perfringens on the production performance of broilers and the damage to the intestinal mucosal structure; it can also improve chicken macrophages. Cellular immune function, can promote broiler growth.

Description

technical field [0001] The invention relates to the screening and application of plant lactobacillus capable of inhibiting Clostridium perfringens. Background technique [0002] Necrotic enteritis (NE) is an acute intestinal infectious disease of poultry mainly caused by type A or type C Gram-positive anaerobic bacteria Clostridium perfringens, and is the most harmful intestinal infection in poultry farming. one of the diseases. Necrotic enteritis was first reported in the UK (Parish et al, 1961), and was subsequently reported by countries all over the world. The occurrence of chicken necrotic enteritis is mainly characterized by severe intestinal bleeding, intestinal mucosal injury, ulcer, necrosis, etc. (Guo S et al, 2015), the disease can occur all year round, especially in hot and rainy summer , mostly occurs in 2-6 week-old broilers and 3-6-month-old commercial layers (Cooper KK et al., 2013, 2010). The occurrence of acute chicken necrotic enteritis can lead to the s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/747A61P1/00A61P31/04A61P37/04C12R1/25
CPCA61K2035/115C12N1/205C12R2001/25
Inventor 李卫芬王佰魁俞国乔吴艳萍毛予龙黄怡付爱坤王阳王一冰
Owner ZHEJIANG UNIV
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