Novel Burkholderia cepacia resistant to preservatives and application of novel Burkholderia cepacia resistant to preservatives

Abstract

The invention discloses a novel Burkholderia cepacia resistant to preservatives and application of the novel Burkholderia cepacia resistant to the preservatives. The new burkholderia cepacia Bcc621 is preserved in the Guangdong Microbial Culture Collection Center (GDMCC), the address is the 5th floor, No. 59 building, No. 100 courtyard, Xianlie Middle Road, Guangzhou, the postal code is 510070, the preservation date is December 17, 2025, and the preservation number is GDMCC No: 67496. The bacterium is resistant to kathon, phenoxyethanol, sodium benzoate, potassium sorbate and 1, 3-dihydroxymethyl-5, 5-dimethyl hydantoin, and the bacterium is resistant to drugs of kathon, phenoxyethanol, sodium benzoate, potassium sorbate and 1, 3-dihydroxymethyl-5, 5-dimethyl hydantoin. According to the present invention, the KirbyBauer (KB) drug sensitive experiment results show that the new Burkholderia cepacia is resistant to the preservative;

Description

Technical Field

[0001] This invention belongs to the field of microbiology, specifically relating to a new strain of preservative-resistant Burkholderia cepacia and its applications. Background Technology

[0002] Burkholderia neonsis ( Burkholderia cenocepacia It is an opportunistic pathogen that causes respiratory infections in patients with cystic fibrosis (CF) and is also one of the bacteria that often cause pollution in industrial production. It has a large genome size (7.8 Mb) and has a natural advantage in adapting to the environment and developing drug resistance. Summary of the Invention

[0003] The purpose of this invention is to provide a new preservative-resistant Burkholderia cepacia strain (… Burkholderia cenocepacia Bcc621 is deposited at the Guangdong Provincial Microbial Culture Collection Center (GDMCC), located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, 510070, China. The deposit date is December 17, 2025, and the accession number is GDMCC No: 67496.

[0004] This invention demonstrates through experiments that *Burkholderia neonata* (…) Burkholderia cenocepacia Bcc621 is resistant to Kathon, phenoxyethanol, sodium benzoate, potassium sorbate and 1,3-dihydroxymethyl-5,5-dimethylhydantoin.

[0005] Therefore, the object of this invention is to provide a new preservative-resistant Burkholderia cepacia strain (… Burkholderia cenocepacia )Bcc621.

[0006] A second objective of this invention is to provide a microbial inoculant containing Burkholderia neonta Bcc621.

[0007] A third objective of this invention is to provide the application of the aforementioned novel Burkholderia cepacia Bcc621 in the study of bacterial resistance mechanisms.

[0008] A fourth objective of this invention is to provide the application of the aforementioned *Burkholderia cepacia* Bcc621 in screening preservatives against *Burkholderia cepacia*.

[0009] Preferably, the screening uses Burkholderia neonavicularis Bcc621 as the model strain for screening against Burkholderia neonavicularis preservatives.

[0010] Preferably, the screening involves preparing the drug to be tested into a drug sensitivity test disc and determining its minimum inhibitory concentration against Burkholderia cepacia Bcc621.

[0011] Preferably, the preservative is Kathon, phenoxyethanol, sodium benzoate, potassium sorbate, or 1,3-dihydroxymethyl-5,5-dimethylhydantoin.

[0012] This invention, through Kirby-Bauer (K-B) susceptibility testing, revealed that *Burkholderia neonavi* Bcc621 is resistant to Kathon, phenoxyethanol, sodium benzoate, potassium sorbate, and 1,3-dimethylol-5,5-dimethylhydantoin, with inhibition zone diameters of 11.88, 14.54, 9.51, 11.55, and 13.22 mm, respectively. Therefore, *Burkholderia neonavi* Bcc621 can be used for resistance research and the development of novel antibacterial agents.

[0013] Burkholderia cenocepacia Bcc621 is deposited at the Guangdong Provincial Microbial Culture Collection Center (GDMCC), located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, 510070, China. The deposit date is December 17, 2025, and the accession number is GDMCC No: 67496. Detailed Implementation

[0014] The following embodiments are further illustrations of the present invention, but not limitations thereof.

[0015] Example 1:

[0016] Burkholderia neonsis ( Burkholderia cenocepacia Bcc621 is a strain isolated from daily chemical products by the Guangdong Provincial Center for Microbiology Analysis and Testing. After the genus was determined by 16S rRNA sequencing, the strain was identified by MLST.

[0017] The 16S rRNA of Burkholderia neon. Bcc621 is shown below:

[0018] Burkholderia neonsis ( Burkholderia cenocepacia Bcc621 is deposited at the Guangdong Provincial Microbial Culture Collection Center (GDMCC), located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, 510070, China. The deposit date is December 17, 2025, and the accession number is GDMCC No: 67496.

[0019] Example 2:

[0020] Burkholderia neonsis ( Burkholderia cenocepacia Preparation of suspension for Bcc621 (abbreviated as Bcc621): A sample of Burkholderia cepacia Bcc621 culture in the exponential growth phase was taken, centrifuged, washed once with PBS buffer, resuspended in PBS, and diluted to 10⁻⁶. 8 CFU / mL was used to obtain a Bcc621 bacterial suspension.

[0021] 1. K‒B susceptibility test for strain Bcc621

[0022] The experimental procedures for the K‒B antimicrobial susceptibility testing disk diffusion method should follow the Clinical and Laboratory Standards Institute (CLSI) 2020 guidelines. Prepare a bacterial suspension by picking a single colony of Bcc621 that has grown overnight from an LB agar plate, and adjust the concentration of the suspension to 1-2 × 10⁻⁶ using physiological saline. 8 CFU·mL -1 Inoculate with the bacterial suspension at the above concentration, dip a sterile cotton swab into the suspension, and spread it evenly on an MH agar plate. Place a drug sensitivity test disc flat in the center of the plate. Incubate at 37 ℃ for 16–18 h, and measure the diameter of the inhibition zone. Interpret the results according to CLSI 2020; the interpretation criteria are detailed in Table 1. The quality control strain used in this experiment was *Burkholderia neonsis* LMG18828.

[0023] Table 1. Antimicrobial Susceptibility Testing Criteria for Burkholderia neonavicularis Bcc621 Kirby-Bauer (K-B)

[0024] Experimental results:

[0025] The drug susceptibility test results for *Burkholderia neocarpa* strain Bcc621 are detailed in Table 2. The results showed that *Burkholderia neocarpa* Bcc621 was resistant to Kathon, phenoxyethanol, sodium benzoate, potassium sorbate, and 1,3-dimethylol-5,5-dimethylhydantoin, with inhibition zone diameters of 11.88, 14.54, 9.51, 11.55, and 13.22 mm, respectively. These results indicate that *Burkholderia neocarpa* Bcc621 is a drug-resistant bacterium resistant to multiple preservatives.

[0026] Table 2. Antimicrobial susceptibility test results of Burkholderia neonavicularis strain Bcc621

Claims

1. Burkholderia neonsis ( Burkholderia cenocepacia Bcc621, accession number: GDMCCNo: 67496.

2. A microbial inoculant, characterized in that, Contains the novel Burkholderia cepacia Bcc621 as described in claim 1.

3. The application of the novel Burkholderia cepacia Bcc621 as described in claim 1 in the study of bacterial resistance mechanisms.

4. The use of Burkholderia cepacia Bcc621 as described in claim 1 in screening preservatives against Burkholderia cepacia.

5. The application according to claim 4, characterized in that, The screening process involved using Burkholderia neonosa Bcc621 as the model strain for screening against Burkholderia neonosa preservatives.

6. The application according to claim 5, characterized in that, The screening process involves preparing the drug to be tested into a drug sensitivity test disc and determining its minimum inhibitory concentration against Burkholderia cepacia Bcc621.

7. The application according to claim 4, characterized in that, The preservatives mentioned are Kathon, phenoxyethanol, sodium benzoate, potassium sorbate, or 1,3-dihydroxymethyl-5,5-dimethylhydantoin.

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