CN108118048A - Regulate and control the method and its application of Salmonella growth - Google Patents

Abstract

The invention discloses a kind of method and its application of regulation and control Salmonella growth, and this method is that salmonella is carried out photo-irradiation treatment, wherein, in the case of the photo-irradiation treatment inhibits the growth of salmonella, the light quality of the photo-irradiation treatment is selected from blue light and/or yellow light；In the case of the photo-irradiation treatment promotes the growth of salmonella, the light quality of the photo-irradiation treatment is selected from green light and/or feux rouges.The method of the regulation and control Salmonella growth has the advantages that radiationless pollution risk, of low cost and easy to operate, and then can be applied in the occasion for needing Salmonella growth.

Description

Method for regulating the growth of Salmonella and its application

technical field

The invention relates to the control of bacteria, in particular to a method for regulating the growth of Salmonella and its application.

Background technique

Salmonella (Salmonella) is a common zoonotic pathogen, which can not only cause typhoid fever and cholera in animals, but also human gastroenteritis, sepsis and other diseases, which seriously threaten the life and health of humans and animals. Food safety incidents caused by it It ranks first among all food-borne pathogenic bacteria. Salmonella has low nutritional requirements and survives for a long time (5-12 months) in food (meat, eggs, milk, bread, etc.), feces, soil, and water, and the contaminated food has no obvious change, so it is easily eaten by animals and humans And cause infectious diseases, animal diseases mainly include fowl typhoid, pullorum, hog cholera, etc. Human infection mainly causes gastroenteritis, typhoid fever, sepsis, cold and other infectious food poisoning. According to statistics released by the World Health Organization in 2015, since 2010, 582 million cases of 22 different foodborne enteric diseases have broken out, causing more than 350,000 deaths, including 52,000 cases of Salmonella typhi and pathogenic E. There were 37,000 cases of Shiella and 35,000 cases of Norovirus. Therefore, Salmonella is the key monitoring object of foodborne pathogens. Therefore, it is particularly important to find effective techniques for inhibiting the growth of Salmonella. Traditional antibacterial methods mainly include chemical drug methods, biological control technology, and physical control technology, as follows:

Traditional antibacterial methods generally use chemical drugs (Li Yang, Ju Yulin, Wang Xiaobo, Liu Ting. Research on the inhibitory effect of traditional Chinese medicine "Lianhuang" on drug-resistant Salmonella[J]. Jilin Animal Husbandry and Veterinary Medicine: 2007,28(9): 9-11 .; Jiang Yunbin, Li Xihong, Fan Xuetong, Tian Guangjuan, Li Limei, Pang Lingling. Effects of atomized plant essential oils on the inhibitory effect of Salmonella typhimurium and the quality of cherry tomatoes[J]. Food Industry Science and Technology, 2017, 09:324-328.) , the effect is obvious, the sterilization is fast, but the process is complicated and the cost is high.

Biological control technology has a good effect on inhibiting the growth of Salmonella, but its process is complicated, the cost of funds is large, and its practical application still has a certain distance (Yan Qigui, Wang Xin, Guo Wanzhu, Yuan Mengwei, Song Zhenhui, Yin Huaping, Li Jing, Cao Hongzhi, Chen Bin. Antagonistic effect of three strains of Bacillus on Salmonella in vitro[J]. Chinese Journal of Preventive Veterinary Medicine, 2007, 29(01):71-74).

Measures to improve the body’s immune system against Salmonella, such as using mannan oligosaccharides or beneficial bacteria to combine with a variety of intestinal pathogens, reduce their adhesion to the intestinal tract and promote excretion, thereby reducing the risk of foodborne diseases (Gaolong. Mannobiose inhibits the adhesion of Salmonella and Escherichia coli [D]. Chinese Academy of Agricultural Sciences, 2016. Li Qing. Screening of lactic acid bacteria inhibiting Salmonella and its protective mechanism on Caco-2 cells [D]. Nanjing Agricultural University, 2015)

Physical control technology is the use of radiation: γ-rays (Ma Haili, Han Keguang, Zheng Mingxue, Li Guozhu, Chai Guizhen, Liu Guanzhang. The radiation effect of 60Co γ-rays on Salmonella in high-immunity egg yolk liquid [J]. Nuclear Agricultural Sciences, 2002, 16 (02) :119-121;) Ultraviolet and collaborative technology (Luo Wei, Application of ultraviolet combined with lactic acid bacteria treatment in the safety control of fresh-cut apples [D]. Sichuan Agricultural University, 2015), high technical equipment requirements or high processing costs, and has a certain The risk of radiation pollution is not suitable for large-scale industrial applications.

Therefore, how to provide a method for regulating Salmonella that has no risk of radiation pollution, low cost, and simple operation is an urgent problem to be solved.

Contents of the invention

The object of the present invention is to provide a method for regulating the growth of Salmonella and its application. The method for regulating the growth of Salmonella has the advantages of no risk of radiation pollution, low cost and simple operation, so that it can be applied to occasions requiring regulation of Salmonella.

In order to achieve the above object, the present invention provides a method for regulating and controlling the growth of Salmonella, the method is to light-treat Salmonella, wherein, under the condition that the light treatment inhibits the growth of Salmonella, the light quality of the light treatment is selected from Blue light and/or yellow light; in case the light treatment promotes the growth of Salmonella, the light quality of the light treatment is selected from green light and/or red light.

The present invention also provides an application of the above-mentioned method for regulating Salmonella in the occasion of regulating Salmonella.

In the above technical solution, the present invention inhibits the growth of Salmonella by adopting light treatment, wherein, when the light quality of the light treatment is blue light and/or yellow light, the light treatment can inhibit the growth of Salmonella; When the light quality is green light and/or red light, the light treatment can promote the growth of Salmonella. The method has the advantages of no risk of radiation pollution, low cost and simple operation, and thus can be applied to occasions that need to regulate Salmonella (such as food preservation).

Other features and advantages of the present invention will be described in detail in the following detailed description.

Description of drawings

The accompanying drawings are used to provide a further understanding of the present invention, and constitute a part of the description, together with the following specific embodiments, are used to explain the present invention, but do not constitute a limitation to the present invention. In the attached picture:

Figure 1 is a graph showing the colony growth of Salmonella in the blue group, yellow group, green group, red group, and CK group.

Detailed ways

Specific embodiments of the present invention will be described in detail below. It should be understood that the specific embodiments described here are only used to illustrate and explain the present invention, and are not intended to limit the present invention.

Neither the endpoints nor any values of the ranges disclosed herein are limited to such precise ranges or values, and these ranges or values are understood to include values approaching these ranges or values. For numerical ranges, between the endpoints of each range, between the endpoints of each range and individual point values, and between individual point values can be combined with each other to obtain one or more new numerical ranges, these values Ranges should be considered as specifically disclosed herein.

The present invention provides a method for regulating the growth of Salmonella. The method is to subject Salmonella to light treatment, wherein, under the condition that the light treatment inhibits the growth of Salmonella, the light quality of the light treatment is selected from blue light and/or Yellow light; in case the light treatment promotes the growth of Salmonella, the light quality of the light treatment is selected from green light and/or red light.

In the above method, the light intensity of blue light can be selected in a wide range, but in order to have a more significant effect on the regulation of Salmonella, preferably, the light intensity of blue light is 750-1550lx;

In the above method, when the light quality of the light treatment contains blue light, in order to have a more significant effect on the regulation of Salmonella, preferably, the blue light also meets the following conditions: the irradiance is 40-50W/m ² , the S/P value 19-21, the correlated color temperature is 90000-110000K, the peak wavelength range is 430-470nm, the half-wave width is 19-22nm, and the dominant wavelength is 430-470nm.

In the above method, the illumination intensity of the yellow light can be selected within a wide range, but in order to have a more significant effect on the regulation of Salmonella, preferably, the illumination intensity of the yellow light is 7500-15300lx;

In the above method, if the light quality of the light treatment contains yellow light, in order to have a more significant effect on the regulation of Salmonella, preferably, the yellow light also meets the following conditions: the irradiance is 30-36W/m ² , S/ The P value is 1-1.2, the correlated color temperature is 3000-4000K, the peak wavelength range is 530-590nm, the half-wave width is 110-115nm, and the dominant wavelength is 550-590nm.

In the above method, the light intensity of the green light can be selected in a wide range, but in order to have a more significant effect on the regulation of Salmonella, preferably, the light intensity of the green light is 3000-6680lx;

In the above method, when the light quality of the light treatment contains green light, in order to have a more significant effect on the regulation of Salmonella, preferably, the green light also meets the following conditions: the irradiance is 10-15W/m ² , S/ The P value is 2-2.5, the correlated color temperature is 6000-6500K, the peak wavelength range is 490-560nm, the half-wave width is 60-70nm, and the dominant wavelength is 540-550nm.

In the above method, the light intensity of the red light can be selected in a wide range, but in order to have a more significant effect on the regulation of Salmonella, preferably, the light intensity of the red light is 900-1930lx;

In the above method, when the light quality of the light treatment contains red light, in order to have a more significant effect on the regulation of Salmonella, preferably, the red light also meets the following conditions: the irradiance is 15-20W/m ² , S/ The P value is 0.1-0.2, the correlated color temperature is 950-1050K, the peak wavelength range is 600-650nm, the half-wave width is 90-100nm, and the dominant wavelength is 600-650nm.

In the above method, the time of light treatment can be selected within a wide range, but in order to have a more significant effect on the regulation of Salmonella, preferably, the time of light treatment is not less than 2 hours.

In order to save cost and improve work efficiency, more preferably, the illumination time is 24-60h.

In the above method, the temperature of the light treatment can be selected within a wide range, but in order to have a more significant effect on the regulation of Salmonella, preferably, the temperature of the light treatment is 5-40°C.

In the above method, there are various ways of providing light for the illumination treatment, but from an economical point of view, preferably, the light for the illumination treatment is provided by an LED light source.

In the present invention, the way of obtaining Salmonella can also be selected in a wide range, but in order to further improve the efficiency of obtaining Salmonella, preferably, before light, the method also includes: Inoculate in LB liquid medium and cultivate for 1-2 days, then expand the cultured bacterial solution and spread it to LB solid medium for cultivation.

The present invention also provides an application of the above-mentioned method for controlling Salmonella in occasions requiring regulation of Salmonella (such as food preservation).

The present invention will be described in detail below by way of examples.

Preparation Example 1

Configuration of LB liquid medium: 10g tryptone, 5g yeast extract, 10g NaCl; weigh the above-mentioned substances according to the liquid medium formula, add 900ml deionized water, shake until the solute is completely dissolved, then use 5mol/L NaOH solution Adjust the pH to 7.0; then dilute to 1L with deionized water; then steam sterilize at 121°C for 20 minutes under high pressure; finally take 100mL of liquid culture medium for later use.

Preparation example 2

The configuration of LB solid medium: 10g tryptone, 5g yeast extract, 10g NaCl, 15g agar powder; weigh the above substances according to the solid medium formula, add 900ml deionized water, heat on the heater until the agar is completely dissolved , use 5mol/L NaOH solution to adjust the pH to 7.0, then dilute to 1L with deionized water; then sterilize by steam at 121°C for 20 minutes, when the temperature drops to about 55°C, dispense while it is hot, generally pour 15ml into 1 After pouring the culture medium into the culture dish, open the lid and expose it to ultraviolet light for 10-15 minutes; finally save it, seal it with sealing glue, and store it upside down at 4°C for later use.

Example 1

1) Take out the Salmonella strains stored in the refrigerator at 4°C and inoculate them in LB liquid medium, and expand the culture for 24 hours.

2) Add 1 ml of the bacterial solution expanded in the second step to 9 ml of sterile water, dilute 10 times, then take 1 ml of the diluted bacterial solution and add it to 9 ml of sterile water, dilute in this way To the 6th power of 10, take 0.1 ml of the bacterial solution diluted 6 times, and spread it into the prepared LB solid medium for culture.

3) Inverted culture was irradiated with the blue light source of blue LED lamp, and the number of colonies was counted by plate colony counting method. Adjust the blue light of the blue LED lamp to the light intensity of 1504lx (the irradiance is 46.6W/m ² , the S/P value is 20.9, the correlated color temperature is 100000K, the peak wavelength is 444nm, the half-wave width is 21nm, and the dominant wavelength is 450nm), It was irradiated at 37°C for 24 hours and recorded as the blue group.

Example 2

Carry out according to the method of embodiment 1, difference is: in step 3) with the yellow light source of yellow LED lamp irradiation upside-down culture, plate colony counting method counts the number of colonies. Use the remote control to adjust the yellow LED light to a light intensity of 15281lx (irradiance is 35.7W/m ² , S/P value is 1.1, correlated color temperature is 3648K, peak wavelength is 551nm, half-wave width is 113nm, and dominant wavelength is 571nm) , cultured at 37°C for 24h, recorded as the yellow group.

Example 3

Carry out according to the method of embodiment 1, difference is: in step 3) in step 3) with the green light source of green LED light irradiation inverted culture, plate colony counting method counts the number of colonies. Use the remote control to adjust the green LED light to the light intensity of 6676lx (irradiance is 14W/m ² , S/P value is 2.2, correlated color temperature is 6304K, peak wavelength is 524nm, half-wave width is 65nm, dominant wavelength is 545nm), Cultivate at 37°C for 24 hours, and record it as the green group.

Example 4

Carry out according to the method of embodiment 1, difference is: in step 3) in the red light source irradiation with red LED lamp inverted culture, plate colony counting method counts the number of colonies. Use the remote control to adjust the red LED light to a light intensity of 1921lx (irradiance is 17W/m ² , S/P value is 0.17, correlated color temperature is 1001K, peak wavelength is 646nm, half-wave width is 95nm, dominant wavelength is 617nm), Incubate at 37°C for 24 hours, and record it as the red group.

Example 5

Carry out according to the method of embodiment 1, difference is: in step 3), the blue light of blue LED lamp is adjusted to light intensity 750lx (irradiance is 40W/m ² , S/P value is 19, correlated color temperature is 90000K, The peak wavelength is 430nm, the half-wave width is 19nm, and the dominant wavelength is 434nm), and it is irradiated at 40°C for 60h.

Example 6

Carry out according to the method of embodiment 1, difference is: in step 3), the blue light of blue LED lamp is adjusted to light intensity 1550lx (irradiance is 50W/m ² , S/P value is 21, correlated color temperature is 110000K, The peak wavelength is 465nm, the half-wave width is 22nm, and the dominant wavelength is 470nm), and it is irradiated at 5°C for 2h.

Example 7

Carry out according to the method of embodiment 1, difference is: in step 3) with the yellow light source of yellow LED lamp irradiation upside-down culture, plate colony counting method counts the number of colonies. Use the remote control to adjust the yellow LED light to a light intensity of 7500lx (irradiance is 30W/m ² , S/P value is 1, correlated color temperature is 3000K, peak wavelength is 530nm, half-wave width is 110nm, dominant wavelength is 550nm) 40 Cultivate for 60h.

Example 8

Carry out according to the method of embodiment 1, difference is: in step 3) with the yellow light source of yellow LED lamp irradiation upside-down culture, plate colony counting method counts the number of colonies. Use the remote control to adjust the yellow LED light to the light intensity of 15300lx (irradiance is 36W/m ² , S/P value is 1.2, correlated color temperature is 4000K, peak wavelength is 589nm, half-wave width is 115nm, dominant wavelength is 590nm), Incubate at 5°C for 2h.

Example 9

Carry out according to the method of embodiment 1, difference is: in step 3) in step 3) with the green light source of green LED light irradiation inverted culture, plate colony counting method counts the number of colonies. Use the remote control to adjust the green LED light to a light intensity of 3000lx (irradiance is 10W/m ² , S/P value is 2, correlated color temperature is 6000K, peak wavelength is 490nm, half-wave width is 60nm, dominant wavelength is 540nm), Incubate at 40°C for 60h.

Example 10

Carry out according to the method of embodiment 1, difference is: in step 3) in step 3) with the green light source of green LED light irradiation inverted culture, plate colony counting method counts the number of colonies. Use the remote control to adjust the green LED light to a light intensity of 6680lx (irradiance is 15W/m ² , S/P value is 2.5, correlated color temperature is 6500K, peak wavelength is 555nm, half-wave width is 70nm, and dominant wavelength is 550nm), Incubate at 5°C for 2h.

Example 11

Carry out according to the method of embodiment 1, difference is: in step 3) in the red light source irradiation with red LED lamp inverted culture, plate colony counting method counts the number of colonies. Use the remote control to adjust the red LED light to a light intensity of 900lx (irradiance is 15W/m ² , S/P value is 0.1, correlated color temperature is 950K, peak wavelength is 600nm, half-wave width is 90nm, dominant wavelength is 600nm), Incubate at 40°C for 60h.

Example 12

Carry out according to the method of embodiment 1, difference is: in step 3) in the red light source irradiation with red LED lamp inverted culture, plate colony counting method counts the number of colonies. Use the remote control to adjust the red LED light to a light intensity of 1930lx (irradiance is 20W/m ² , S/P value is 0.2, correlated color temperature is 1050K, peak wavelength is 650nm, half-wave width is 100nm, and dominant wavelength is 650nm), Incubate at 5°C for 2h.

Comparative example 1

Carry out according to the method of Example 1, the difference is that in step 3): irradiate with natural light and culture upside down, count the number of colonies by plate colony counting method. Cultured at 37°C for 24 hours, and recorded as CK group.

Result analysis

1) The above-mentioned Salmonella colony growth results of the blue group, yellow group, green group, red group, and CK group are shown in Figure 1 (the first row in the figure is the yellow group, the red group, and the CK group from left to right, and the second row is composed of Left to right are green group and blue group respectively); As can be seen from Figure 1, the colonies of the control group (CK group) grow well and have a large colony area. Compared with the CK group, the green group, red group, yellow group, and blue group have different colony growth conditions, among which the green light and red light groups can promote the growth of Salmonella, and the yellow light and blue light groups have obvious inhibitory ability to Salmonella , in which the antibacterial ability of the blue light group was stronger than that of the yellow light.

2) Effect of light on the growth of Salmonella colony

After culturing according to the culture method, the number of colonies in each group was determined by plate counting method, and the significant difference was analyzed by SPSS13.0 software. The results are shown in Table 1.

Table 1

blue team yellow group green group red group CK group Colony count (10 ⁷ CFU/mL) 0 39±15 177±32 167±26 116±27

The colony germination of the treatment group and the control group was significantly different, among which the blue group<yellow group<CK group<red group≈green group. According to SPSS analysis, there were extremely significant differences between the treatment group and the control group, and the blue group was about CK 1/100 of the group, the yellow group is about 1/4 of the CK group, and the green group and red group are about 1.3. Except for green light and red light, there was no significant difference among the experimental groups, and there were extremely significant differences among the other groups.

Embodiment 5-12 is detected according to the same method, wherein, the detection result of embodiment 5-6 is basically consistent with the blue group, the detection result of embodiment 7-8 is basically consistent with the yellow group, and the detection result of embodiment 9-10 is consistent with the green group. The detection results of the red group are basically the same, and the detection results of Examples 11-12 are basically the same as those of the red group.

The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention. These simple modifications All belong to the protection scope of the present invention.

In addition, it should be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable way if there is no contradiction. The combination method will not be described separately.

In addition, various combinations of different embodiments of the present invention can also be combined arbitrarily, as long as they do not violate the idea of the present invention, they should also be regarded as the disclosed content of the present invention.

Claims (10)

1. a method for regulating and controlling the growth of Salmonella, is characterized in that, described method is that Salmonella is carried out light treatment; Wherein, in the case that the light treatment inhibits the growth of Salmonella, the light quality of the light treatment is selected from blue light and/or yellow light; in the case that the light treatment promotes the growth of Salmonella, the light quality of the light treatment The quality is selected from green light and/or red light. 2. The method according to claim 1, wherein the illumination intensity of the blue light is 750-1550lx; Preferably, the blue light also meets the following conditions: the irradiance is 40-50W/m ² , the S/P value is 19-21, the correlated color temperature is 90000-110000K, the peak wavelength range is 430-470nm, and the half-wave width is 19 -22nm, the dominant wavelength is 430-470nm. 3. The method according to claim 1, wherein the illumination intensity of the yellow light is 7500-15300lx; Preferably, the yellow light also meets the following conditions: the irradiance is 30-36W/m ² , the S/P value is 1-1.2, the correlated color temperature is 3000-4000K, the peak wavelength range is 530-590nm, and the half-wave width is 110-115nm, the dominant wavelength is 550-590nm. 4. The method according to claim 1, wherein the illumination intensity of the green light is 3000-6680lx; Preferably, the green light also meets the following conditions: the irradiance is 10-15W/m ² , the S/P value is 2-2.5, the correlated color temperature is 6000-6500K, the peak wavelength range is 490-560nm, and the half-wave width is 60-70nm, the dominant wavelength is 540-550nm. 5. The method according to claim 1, wherein the illumination intensity of the red light is 900-1930lx; Preferably, the red light also meets the following conditions: the irradiance is 15-20W/m ² , the S/P value is 0.1-0.2, the correlated color temperature is 950-1050K, the peak wavelength range is 600-650nm, and the half-wave width is 90-100nm, the dominant wavelength is 600-650nm. 6. The method according to any one of claims 1-5, wherein the time of the light treatment is not less than 2h. 7. The method according to any one of claims 1-5, wherein the temperature of the light treatment is 5-40°C. 8. The method according to any one of claims 1-5, wherein said light treatment light is provided by an LED light source. 9. According to any one of claims 1-5, wherein, before the illumination, the method further comprises: first inoculating the Salmonella bacterial strains preserved at 3-5°C in LB liquid medium and culturing 1- After 2 days, the expanded cultured bacterial solution was diluted and applied to LB solid medium for cultivation. 10. An application of the method for regulating Salmonella according to any one of claims 1-8 in the field of regulating Salmonella.