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A cold-water fish probiotic Lactococcus lactis strain and its application

A technology of Lactococcus lactis and cold-water fish, applied in the field of microbiology, can solve the problem of low antibody titer

Inactive Publication Date: 2021-02-19
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Fish have the ability of immune memory response. If you want to obtain a strong immune response again, it will take a long time, and the antibody titer produced by the secondary response is lower than that of mammals.

Method used

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  • A cold-water fish probiotic Lactococcus lactis strain and its application
  • A cold-water fish probiotic Lactococcus lactis strain and its application
  • A cold-water fish probiotic Lactococcus lactis strain and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Isolation, purification and identification of Lactococcus lactis HZC32 strain of the present invention

[0032] GM17 solid medium formula (g / L): soybean peptone 5.0, peptone 2.5, casein peptone 2.5, yeast extract powder 2.5, beef powder 5.0, lactose 5.0, sodium ascorbate 0.5, β-sodium glycerophosphate 19.0, magnesium sulfate 0.25, agar 15. After dissolving, sterilize at 110°C for 15 minutes, adjust the pH to 7.2, and pour into a plate for later use.

[0033] GM17 liquid medium formula (g / L): soybean peptone 5.0, peptone 2.5, casein peptone 2.5, yeast extract powder 2.5, beef powder 5.0, lactose 5.0, sodium ascorbate 0.5, β-sodium glycerophosphate 19.0, magnesium sulfate 0.25, dissolved Afterwards, sterilize at 110°C for 15 minutes, adjust the pH to 7.2, and set aside.

[0034] (1) Isolation and purification of bacterial strains

[0035] To collect experimental juvenile fish from the natural and pollution-free natural waters of Benxi, Liaoning, the stress of ...

Embodiment 2

[0041] Example 2 Research on the psychrophilic characteristics of Lactococcus lactis HZC32 strain of the present invention

[0042] The unique psychrophilic properties of the Lactococcus lactis HZC32 strain:

[0043] (1) Experimental method: Prepare solid and liquid GM17 medium, streak the Lactococcus lactis HZC32 bacterial liquid on the medium, and set 5 culture temperatures of 14°C, 20°C, 25°C, 30°C and 37°C respectively. Cultured and observed continuously for 4 days.

[0044] (2) Experimental results: It grows well between 14°C and 18°C ​​(see Table 1), grows well on solid and liquid GM17 medium, and the colony on GM17 medium is milky white, with neat edges, smooth and moist surface, and raised , opaque large colonies, with a peculiar sour or cheesey smell; the bacteria has typical psychrophilic characteristics: it can grow at 4°C, grow well between 14°C and 18°C, and does not grow above 20°C.

[0045] Table 1

[0046] temperature 4℃ 14℃ 18℃ 20℃ Strain...

Embodiment 3

[0047] Example 3 Research on the acid resistance of Lactococcus lactis HZC32 strain of the present invention

[0048] The unique acid resistance characteristics of the Lactococcus lactis HZC32 strain:

[0049] (1) Experimental method: The Lactococcus lactis HZC32 strain was cultured at 15°C, and the strains were observed in GM17 liquid medium with different pH (pH3.0, pH4.0, pH5.0, pH6.0 and pH6.5). of growth.

[0050] (2) Experimental results: the Lactococcus lactis HZC32 strain has good adaptability in media of different pH, and can proliferate in the acidic GM17 medium when the pH is 4.0; it can proliferate in the acidic GM17 medium with a pH of 5.0 Faster; proliferates rapidly in near-neutral GM17 medium with a pH value of 6.5. See figure 2 The proliferation curve indicated that the Lactococcus lactis HZC32 has the ability to tolerate the low pH environment of the intestinal tract, and the Lactococcus lactis HZC32 strain has the ability to colonize in the acidic intest...

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Abstract

The invention discloses a cold-water fish probiotic Lactococcus lactis strain and its application. The present invention isolates and identifies a strain HZC32 that has antiviral properties, immune regulation and immune function enhancement properties to cold water fish from the intestinal mucosa of semi-wild rainbow trout juveniles in the Benxi mountainous area of ​​Liaoning Province. According to the observation of the morphological characteristics of the colony and the determination of the 16SrDNA base sequence and homology analysis, the strain identification result is: HZC32 belongs to Lactococcus lactis, and the preservation number is: CGMCC No.10705. The in vitro cell experiments proved that: Lactococcus lactis HZC32 strain co-cultured with virus-infected EPC cells has the property of resisting IHNV proliferation in EPC cells in vitro. Animal experiments have proved that: Lactococcus lactis HZC32 strain has the characteristics of immune regulation and immune function enhancement to cold-water fish, and the HZC32 strain fed alone also has the characteristics of anti-IHNV infection to cold-water fish. Thus it can be seen that the Lactococcus lactis HZC32 strain of the present invention has a certain application prospect in cold-water fish anti-virus, immune regulation and enhancement of cold-water fish immune function.

Description

technical field [0001] The invention relates to a probiotic bacterial strain and its use, in particular to a cold-water fish (rainbow trout) probiotic Lactococcus lactis HZC32 strain and its use. The invention belongs to the technical field of microbes. Background technique [0002] Probiotics are the normal flora of human and animal intestinal tract, respiratory tract, reproductive tract and skin mucous membrane system. Many research results have found that lactic acid bacteria and intestinal mucosa together form a protective barrier to prevent the invasion of viruses and bacteria. Human beings are increasingly aware of In view of the powerful effect of probiotics on human and animal life and health, a large number of modern immunological research results have confirmed that probiotics (especially lactic acid bacteria) can regulate animal cellular immunity, humoral immunity and local immunity of intestinal mucosa, as well as immune system development and The indispensable d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K50/80A23K10/18A61K39/39A61K39/205A61K35/744A61K39/385A61P31/14A61P37/02A61P37/04C12R1/01
CPCY02A40/818
Inventor 崔红玉王云峰王笑梅卢彤岩
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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