Two multidrug-resistant, biofilm-forming animal pathogenic bacteria and their applications

A technology of animals and scented bacteria, applied in the field of microorganisms, can solve the problems of reducing the treatment effect of antibiotics, and achieve the effect of strengthening the ability to form biofilms

Active Publication Date: 2021-11-30
GUANGDONG BOWOTE BIOTECH CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the other hand, some pathogenic bacteria that cause diarrhea in pigs have the ability to form biofilms, which greatly reduces the effect of antibiotic treatment

Method used

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  • Two multidrug-resistant, biofilm-forming animal pathogenic bacteria and their applications
  • Two multidrug-resistant, biofilm-forming animal pathogenic bacteria and their applications
  • Two multidrug-resistant, biofilm-forming animal pathogenic bacteria and their applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Isolation, purification and preservation of bacterial strains WP-1 and NP-2

[0031] Collect fresh feces from weaned piglets and nursery pigs, immediately put them into anaerobic sampling tanks, seal them, and bring them back to the laboratory as soon as possible. Take 1g of feces sample, add it to 30mL PBS, and then serially dilute to 10 -8 concentration. take 10 -6 、10 -7 and 10 -8 100 μL of each dilution was coated on a BHI (Qingdao Haibo) plate. At 37°C, observe after 2 days of culture. Colonies were picked, transferred and streaked on a fresh BHI plate for purification 3 times. Strains WP-1 and NP-2 were thus obtained

[0032] WP-1 and NP-2 were cultured using BHI liquid medium. The bacterial cells in the logarithmic growth phase were collected, resuspended in 20% (v / v) glycerol solution, and frozen in a -80°C refrigerator. At the same time, freeze-dried tubes of WP-1 and NP-2 were prepared by Guangdong Microbial Culture Collection Center and st...

Embodiment 2

[0033] Embodiment 2: bacterial strain WP-1 and NP-2 morphological description

[0034] On the BHI plate (Qingdao Haibo), after 48 hours of culture, the colony of strain WP-1 is white in color, about 0.5mm in diameter, round, smooth, opaque, uniform in texture, convex in the center, and neat in edge ( figure 1 ).

[0035] On the BHI plate (Qingdao Haibo), after 48 hours of culture, the colony of strain NP-2 is yellow in color, about 0.5mm in diameter, round, smooth, opaque, uniform in texture, convex in the center, and neat in edge ( figure 2 ).

[0036] Resuspend WP-1 and NP-2 monoclonals in logarithmic growth phase with sterile water, respectively. The cell morphology of WP-1 and NP-2 was observed and photographed using a transmission electron microscope.

[0037] The cell morphology of WP-1 and NP-2 were as follows image 3 and Figure 4 shown.

Embodiment 3

[0038] Example 3: Classification and identification of bacterial strains WP-1 and NP-2

[0039] The 16S rRNA genes of WP-1 and NP-2 were amplified using universal primers 27F and 1492R, and sent to Shanghai Meiji Biomedical Technology Co., Ltd. for sequencing. The nucleotide sequence of the 16S rRNA gene of strain WP-1 is shown in SEQ ID NO .1, the nucleotide sequence of the 16S rRNA gene of bacterial strain NP-2 is shown in SEQ ID NO.2.

[0040] The obtained two-terminal sequencing results were assembled. Compare in EZBioCloud database. Strain WP-1 and model bacteria Myroides odoratus DSM 2801 T It has the highest 16S rRNA gene similarity of 98.4%. Strain NP-2 and model bacteria Myroides odoratus DSM 2801 T It has the highest 16S rRNA gene similarity of 98.5%. The 16S rRNA gene similarity between strains WP-1 and NP-2 was 98.9%.

[0041] Genomic DNA of WP-1 and NP-2 was extracted. Sent to Shanghai Passino Biotechnology Co., Ltd. for sequencing using the IlluminaNovaSeq...

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Abstract

The invention discloses two kinds of animal pathogenic bacteria with multi-drug resistance and biofilm formation and application thereof. The said malodorous aroma bacterium (Myroidesfetidus) WP-1, its preservation number: GDMCC No: 61556. Said yellow flavor fungus (Myroidesflavus) NP‑2, its preservation number: GDMCCNo: 61557. The present invention isolates two new strains of pathogenic bacteria from pig feces with diarrhea, analyzes their drug resistance to 23 kinds of antibiotics, evaluates their ability to form biofilm, and tests their pathogenicity to experimental animals. The invention result has great significance for the detection, diagnosis and treatment of pig diarrheal disease.

Description

Technical field: [0001] The invention belongs to the technical field of microbes, and in particular relates to two kinds of animal pathogenic bacteria with multi-drug resistance and biofilm formation and applications thereof. Background technique: [0002] Pig diarrhea is a common multifactorial disease in veterinary clinics. It often affects pig farms, whether it is a modern large-scale farm or a free-range farm. Its characteristics are mostly manifested as: vomiting, diarrhea, dehydration and so on. Neglect of treatment or untimely treatment will cause death or poor prognosis of pigs, causing great economic losses to farmers and seriously threatening the healthy and sustainable development of the breeding industry. Therefore, the disease is often regarded as the top priority of disease prevention and control in the process of pig breeding. [0003] The pathogenesis of swine diarrhea is relatively complicated, and there are more than a dozen pathogenic factors. At present...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12Q1/04A61K35/74A61P1/10C12R1/01
CPCA61K35/74A61P1/10C12Q1/04G01N2333/195G01N2800/06
Inventor 李安章朱红惠徐帅帅陈猛胡艳娜姚青
Owner GUANGDONG BOWOTE BIOTECH CO LTD
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