Solid beverage with lead ion emission reduction function and application thereof

Abstract

The invention relates to the technical field of microorganisms, in particular to a solid beverage with a lead ion emission reduction function and application of the solid beverage. The preparation method of the solid beverage provided by the invention comprises the following steps: adding the fermented lactobacillus mucus B44 into skimmed milk, uniformly mixing, and drying to obtain the solid beverage. Compared with a control group and other solid beverages prepared by adding lactobacillus, the fermented lactobacillus mucus solid beverage provided by the invention can significantly reduce the lead concentration in the in-vitro intestinal content supernatant, and results of in-vivo experiments of rats show that the fermented lactobacillus mucus solid beverage can significantly reduce the blood lead concentration of the rats suffering from lead poisoning, and has a good application prospect. The lead content in the liver and the lead content in the kidney can be increased, discharge of lead in excrement can be increased, and kidney oxidative damage caused by lead poisoning can be effectively relieved.

Description

Technical Field

[0001] The invention relates to the field of beverages, in particular to a solid beverage capable of reducing lead ion emission and application thereof. Background Art

[0002] Lead pollution is a widespread heavy metal pollution problem. Unlike organic pollutants, these pollutants are not easy to decompose and volatilize, and they are highly persistent in the environment. They may pose a major threat to human health through accumulation in the food chain. Children, in particular, are more sensitive to lead, which may lead to negative consequences such as behavioral problems, inattention, and developmental disorders. Currently, methods to reduce lead levels in the body include the use of traditional chelating agents (such as dimercaprol, calcium EDTA, etc.), as well as nutritional supplementation strategies. However, since chelating agents may bring greater side effects and safety issues, they are not recommended as a treatment for specific groups such as children and the elderly. Therefore, there is an urgent need to develop a safe and effective new method to reduce blood cadmium levels in people with high cadmium exposure in my country.

[0003] Beneficial bacteria are bacteria that are beneficial to the human body. They help maintain the balance of intestinal microecology and have many functions such as promoting intestinal health and improving immunity. There are a wide variety of probiotic products on the market, including fermented dairy products and powdered beverages. Therefore, by taking powdered beverages containing probiotics, the potential problems caused by using traditional chelating agents to reduce heavy metal lead in the body can be avoided. Summary of the invention

[0004] In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to provide a solid beverage capable of reducing the emission of lead ions and its application, so as to solve the problems in the prior art.

[0005] To achieve the above-mentioned object and other related objects, the present invention provides a solid beverage capable of reducing lead ion emission. The solid beverage comprises fermented Lactobacillus mucosa B44 or its active substance, and the fermented Lactobacillus mucosa B44 has a preservation number of CGMCC No.17321.

[0006] Preferably, the active substance is a fermentation culture product of Lactobacillus mucilaginosus B44 with a deposit number of CGMCC No.17321.

[0007] The present invention also provides the use of the solid beverage in preparing a product for reducing lead toxicity.

[0008] The present invention also provides a method for preparing the aforementioned solid beverage, the preparation method comprising the following steps: adding fermented mucus lactobacillus B44 into skim milk and mixing, freeze-drying to obtain a B44 bacterial agent, and then adding sterile skim powder to dilute to obtain a solid beverage.

[0009] As described above, the solid beverage capable of reducing lead ion emission and its application of the present invention have the following beneficial effects:

[0010] The fermented mucus lactobacillus solid beverage provided by the present invention can significantly reduce the lead concentration in the supernatant of intestinal contents in vitro compared with a control group and other solid beverages prepared by adding lactobacillus. According to the results of in vivo experiments on rats, the blood lead concentration, the lead content in the liver and the lead content in the kidney of lead-poisoned rats can be significantly reduced, and the discharge of lead in feces can be increased and the oxidative damage to the kidney caused by lead poisoning can be effectively alleviated. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 Shown are the test results of kidney antioxidant indexes in SD rats after lead poisoning and intervention in the present invention. DETAILED DESCRIPTION

[0012] The invention provides a solid beverage capable of reducing the emission of lead ions. The solid beverage comprises Limosilactobacillus fermentum B44 or an active substance thereof.

[0013] The fermented mucus Lactobacillus B44 is deposited in the General Microbiology Center (CGMCC) of the China Microbiological Culture Collection Administration, with a deposit number of CGMCC No. 17321, a deposit date of 2019-03-08, and a deposit address of the Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.

[0014] In some specific embodiments, the active substance is a fermentation culture product of the fermented mucus lactobacillus B44 with a deposit number of CGMCC No. 17321. Specifically, the fermentation culture product can be selected from a fermentation culture broth containing bacterial cells, a fermentation supernatant after removing bacterial cells, or an active substance extracted from a fermentation culture broth. The fermented mucus lactobacillus B44 of the present invention can be cultured by anaerobically fermenting in a lactobacillus culture medium commonly used in the field (e.g., MRS culture medium). The optimal fermentation temperature is 35-38°C, and the optimal fermentation time is 7-24h.

[0015] In some specific embodiments, based on the mass of the solid beverage, the viable count of Lactobacillus mucilaginosus B44 in the solid beverage is 5×10 10 -2x10 11 CFU / g. Specifically, the viable count is 5x10 10 -7x10 10 CFU / g, 7x10 10 -9x10 10 CFU / g, 9x10 10 -1x10 11 CFU / g or 1x1011 -2x10 11 CFU / g.

[0016] In some specific embodiments, the solid beverage further contains one or more of water-soluble dietary fiber, sweeteners, vitamins, minerals, edible flavors, edible colorings, and preservatives.

[0017] In some specific embodiments, the water-soluble dietary fiber can be selected from one or more of resistant dextrin, inulin, pectin, guar gum, xanthan gum, oligofructose, hyaluronic acid, chitosan or soybean fiber.

[0018] In some specific embodiments, the sweetener can be selected from one or more of honey, lactitol, sucrose, sucralose, fructose, glucose, erythritol, xylitol, corn syrup or fructose syrup.

[0019] In some specific embodiments, the vitamin can be selected from one or more of vitamin C, vitamin A, vitamin D, vitamin E, or vitamin B1, B2, B3, B5, B6, B7, B9, and B12.

[0020] In some specific embodiments, the mineral may be selected from one or more of sodium salts, potassium salts, magnesium salts, calcium salts or phosphates.

[0021] In some embodiments, the preservative can be selected from one or more of benzoic acid or its salts, sorbic acid or its salts, p-hydroxybenzoic acid esters or its salts, or dehydroacetic acid or its salts.

[0022] In some specific embodiments, based on the total weight of the solid beverage, the fermented mucus lactobacillus B44 or its active substance accounts for 0.1-50% of the mass percentage of the solid beverage. Specifically, the mass percentage is 0.1-1%, 1-10%, 10-15%, 15-20%, 20-25%, 25-30%, 30-35%, 35-40% or 40-50%. The present invention also provides the use of the above-mentioned solid beverage in the preparation of a product to reduce lead toxicity.

[0023] In some embodiments, the lead toxicity reduction product has any one or more of the following functions:

[0024] 1) Reduce liver MDA content;

[0025] 2) Increase CAT enzyme activity;

[0026] 3) increasing the GSH content, wherein the reduction or increase is a reduction or increase in the MDA content, CAT enzyme activity or GSH content relative to the lead model.

[0027] The solid beverage provided by the present invention has an effect of improving the removal of lead ions in the supernatant of intestinal contents by ≥28.5% compared with the control group, reduces the blood lead concentration in the lead model SD rats by 49.7% compared with the model group, reduces the lead content in the liver and kidney of the lead model SD rats by 44.3% and 24.3% respectively compared with the control group, and has the effect of promoting the excretion of lead through feces of the lead model SD rats.

[0028] The present invention also provides a method for preparing the solid beverage, the method comprising the following steps:

[0029] The fermented mucus lactobacillus B44 is added into skim milk and mixed evenly, and the mixture is freeze-dried to obtain the B44 bacterial agent, and then sterile skim powder is added and diluted to obtain a solid beverage.

[0030] In some embodiments, based on the volume of skim milk after mixing, the number of viable bacteria of fermented Lactobacillus mucilaginosus B44 is 1×10 7 -1x10 9 CFU / mL. Specifically, the viable count is 1x10 7 -5x10 7 CFU / mL, 5x10 7 -1x10 8 CFU / mL, 1x10 8 -5x10 8 CFU / mL or 5x10 8 -1x10 9 CFU / mL.

[0031] In some embodiments, the skim milk comprises skim milk powder and water.

[0032] Furthermore, based on the mass of skim milk, the mass percentage of skim milk powder to the total mass of skim milk is 5-15%. Specifically, the mass percentage is 5-6%, 6-7%, 7-8%, 8-9%, 9-10%, 10-11%, 11-12%, 12-13%, 13-14% or 14-15%.

[0033] Further, based on the mass of skim milk, the mass percentage of water in the total mass of skim milk is 85-95%. Specifically, the mass percentage is 85-86%, 86-87%, 87-88%, 88-89%, 89-90%, 90-91%, 91-92%, 92-93%, 93-94% or 94-95%.

[0034] In the present invention, "fixed beverage" generally refers to a solid product made from sugar, milk and dairy products, eggs or egg products, fruit juice or edible plant extracts as main raw materials, with appropriate amount of auxiliary materials or food additives added, and the water content of each 100 grams of the finished product is not higher than 5 grams.

[0035] In some embodiments, the drying method can be selected from one or more of freeze drying, vacuum drying or fluidized bed drying.

[0036] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0037] Before further describing the specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the examples of the present invention are for describing the specific embodiments rather than for limiting the scope of protection of the present invention; in the present specification and claims, unless otherwise expressly stated herein, the singular forms "a", "an" and "the" include plural forms.

[0038] When the embodiments give numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those generally understood by those skilled in the art. In addition to the specific methods, equipment, and materials used in the embodiments, according to the grasp of the prior art by those skilled in the art and the record of the present invention, any methods, equipment, and materials of the prior art similar or equivalent to the methods, equipment, and materials described in the embodiments of the present invention can also be used to realize the present invention.

[0039] Example 1

[0040] 1. Preparation of fermented mucus lactobacillus B44 bacterial agent

[0041] The lyophilized powder of fermented mucus lactobacillus B44 was dissolved with a small amount of sterile distilled water, and a loop was streaked on MRS solid medium (purchased from Qingdao High-tech Industrial Park Haibo Biotechnology Co., Ltd., China) with an inoculation loop, and anaerobically cultured at 37°C for 48 hours. A single colony was picked up with an inoculation loop and placed in 5 mL sterile MRS liquid medium (purchased from Qingdao High-tech Industrial Park Haibo Biotechnology Co., Ltd., China). The colony was evenly dispersed in the liquid medium using a vortex oscillator, cultured at 37°C for 24 hours, and inoculated into MRS liquid medium with an inoculum of 3% (v / v). After culturing again at 37°C for 24 hours, the culture was centrifuged at 9,000 rpm and 7°C for 15 minutes, the supernatant was discarded, and the culture was suspended with a small amount of sterile skim milk (15%, v / w). After freeze-drying, the bacterial agent of fermented mucus lactobacillus B44 was obtained. The concentration of the bacterial agent was determined to be 1.5×10 11 CFU / g.

[0042] 2. Preparation of fermented mucus lactobacillus B44 solid beverage

[0043] The fermented Lactobacillus mucilaginosus B44 inoculum was appropriately diluted with sterile skim milk powder and fully mixed to obtain the fermented Lactobacillus mucilaginosus solid beverage A of this embodiment. After testing, the viable count of the fermented Lactobacillus mucilaginosus solid beverage A was 4.0x10 10 CFU / g.

[0044] Example 2

[0045] 1. Preparation of fermented Lactobacillus mucilaginosus B44 bacterial agent: same as Example 1.

[0046] 2. Preparation of fermented mucus lactobacillus B44 solid beverage

[0047] Will Example 1 The fermented Lactobacillus mucilaginosus B44 inoculum was appropriately diluted with sterile skim milk powder and fully mixed to obtain the fermented Lactobacillus mucilaginosus solid beverage B of this embodiment. After testing, the viable count of the fermented Lactobacillus mucilaginosus solid beverage B was 6.0x10 9 CFU / g.

[0048] Example 3

[0049] 1. Preparation of fermented Lactobacillus mucilaginosus B44 bacterial agent: same as Example 1.

[0050] 2. Preparation of fermented mucus lactobacillus B44 solid beverage

[0051] Will Example 1The fermented Lactobacillus mucilaginosus B44 inoculum was appropriately diluted with sterile skim milk powder and fully mixed to obtain the fermented Lactobacillus mucilaginosus solid beverage C of this embodiment. After testing, the viable count of the fermented Lactobacillus mucilaginosus solid beverage C was 8.0×10 9 CFU / g.

[0052] Example 4

[0053] 1. Preparation of fermented Lactobacillus mucilaginosus B44 bacterial agent: same as Example 1.

[0054] 2. Preparation of fermented mucus lactobacillus B44 solid beverage

[0055] Will Example 1 The fermented Lactobacillus mucilaginosus B44 inoculum was appropriately diluted with sterile skim milk powder and fully mixed to obtain the fermented Lactobacillus mucilaginosus solid beverage D of this embodiment. After testing, the viable count of the fermented Lactobacillus mucilaginosus solid beverage D was 1.0×10 10 CFU / g.

[0056] Example 5

[0057] 1. Preparation of fermented Lactobacillus mucilaginosus B44 bacterial agent: same as Example 1.

[0058] 2. Preparation of fermented mucus lactobacillus B44 solid beverage

[0059] Will Example 1 The fermented Lactobacillus mucilaginosus B44 inoculum was appropriately diluted with sterile skim milk powder and fully mixed to obtain the fermented Lactobacillus mucilaginosus solid beverage E of this embodiment. After testing, the viable count of the fermented Lactobacillus mucilaginosus solid beverage E was 5.0×10 10 CFU / g.

[0060] Effect Example 1: Effect of fermented mucus lactobacillus solid beverage on reducing lead ion emission in intestinal contents

[0061] (a) In vitro preparation of intestinal contents

[0062] Feces of a male aged 25 to 35 years old was randomly collected, 25 g of fecal sample was weighed, 225 mL of deionized water was added, and an in vitro intestinal content solution was prepared at a solid-liquid ratio of 1:9 (w / v), which was mixed and set aside.

[0063] (b) Experiment on reduction of lead ions in intestinal contents

[0064] Take 0.1g of each of the fermented mucus lactobacillus solid beverages A to E of Examples 1 to 5 above (the blank group was without any treatment; the sterile skim milk powder was the control group), and mix them with 20mL of the intestinal content solution containing lead ions, so that the final lead ion concentration is 50mg / L. After oscillating at 37°C and 200rpm for 1h, centrifuge at 9,000rpm and 7°C for 15min, and collect the supernatant solution. Take 5mL of the above supernatant solution, add 3mL of concentrated nitric acid (superior purity) and digest it in a graphite digester at 130°C for 30min, rinse the tube twice with deionized water, make the volume to 15mL, and measure it using an inductively coupled plasma optical emission spectrometer (ICP-OES). The specific results are as follows:

[0065] Table 1 Effect of the fermented mucus lactobacillus solid beverage of Examples 1 to 5 on the removal of lead ions in intestinal contents

[0066]

[0067] As can be seen from the above table, intestinal contents have a certain adsorption effect on lead ions in water, resulting in the lead content in the intestinal contents supernatant of the blank group (without any treatment) being lower than the lead content in the initially set solution system. This is due to the effect of bacteria and organic matter in the intestinal contents on lead adsorption; and the control group (sterile skim milk powder) itself has a very limited effect on reducing lead ions in the intestine. However, when fermented mucus lactobacillus B44 is added to the solid beverage, the removal effect of lead is further improved. Specifically, the removal effect of lead ions in the supernatant of intestinal contents by the fermented mucus lactobacillus solid beverage is ≥30.6% higher than that of the blank group, and ≥28.5% higher than that of the control group.

[0068] Effect Example 2 Experimental study on the storage of solid beverage made from fermented Lactobacillus mucilaginosus at room temperature

[0069] The fermented mucus Lactobacillus solid beverages A to E of Examples 1 to 5 were stored at room temperature (25° C.) and taken out at 0, 3 and 6 months. The lead ion concentration in the supernatant samples of each intestinal content was determined by referring to the method described in Effect Example 1. The results are shown in the following table.

[0070] Table 2 Effect of fermented mucus Lactobacillus solid beverages A to E on the removal of lead ions from intestinal contents during storage

[0071]

[0072]

[0073] As can be seen from the above table, after the fermented mucus lactobacillus solid beverage prepared by the present invention is stored at room temperature (25° C.) for 6 months, the lead emission reduction effect decreases slightly in the first 3 months, but the overall lead emission reduction effect remains stable.

[0074] Comparative Example 1

[0075] With reference to the method described in Example 1, the removal effect of lead ions in intestinal contents of solid beverages containing lactobacillus prepared by adding fermented mucus Lactobacillus B44 (provided by Bright Dairy Co., Ltd., Shanghai, China) and Lactobacillus casei ATCC334 (purchased from ATCC, the United States) was compared, and the specific operations were as follows.

[0076] 1. Materials and methods

[0077] (a) Preparation of fermented Lactobacillus mucilaginosus B44 bacterial agent: Same as Example 1

[0078] (b) Preparation of Lactobacillus casei ATCC334 bacterial agent

[0079] The lyophilized powder of Lactobacillus casei ATCC334 was dissolved in a small amount of sterile distilled water, and a loop was streaked on MRS solid medium (purchased from Qingdao High-Tech Industrial Park Haibo Biotechnology Co., Ltd., China) with an inoculation loop, and anaerobically cultured at 37°C for 48 h. A single colony was picked with an inoculation loop and placed in 5 mL of sterile MRS liquid medium (purchased from Qingdao High-Tech Industrial Park Haibo Biotechnology Co., Ltd., China), and the colony was evenly dispersed in the liquid medium using a vortex oscillator, and cultured at 37°C.

[0080] After culturing for 24 hours, the culture was taken out and inoculated into MRS liquid medium at a 3% (v / v) inoculation amount. After culturing again at 37°C for 24 hours, the culture was centrifuged at 9,000 rpm and 7°C for 15 minutes, the supernatant was discarded, and the culture was suspended with a small amount of skim milk (10%, v / w). After freeze-drying, the bacterial agent of Lactobacillus casei ATCC334 was obtained. The concentration of the bacterial agent was determined to be 2.0x10 11 CFU / g.

[0081] 2. Preparation of Lactobacillus solid beverage

[0082] The fermented Lactobacillus mucilaginosus B44 and Lactobacillus casei ATCC334 were appropriately diluted with sterile skim milk powder and fully mixed to obtain solid beverages prepared by adding different lactobacilli. After testing, the number of live bacteria in the above lactobacillus solid beverages was 5.0x10 10 CFU / g.

[0083] 3. Comparison of the reduction effect of lead ions in intestinal contents

[0084] The removal effects of the two lactobacillus solid beverages on lead ions in the supernatant of intestinal contents were determined by the method described in Reference Effect Example 1. The results are shown in the following table.

[0085] Table 3 Effect of different lactobacillus solid beverages on the removal of lead ions in intestinal contents

[0086] Lactobacillus formula milk blank Comparison B44 ATCC334 Lead ion concentration in supernatant (mg / L) 3.6 3.5 1.6 3.0

[0087] By comparison, it can be seen that the removal effect of lead ions in intestinal contents by solid beverages containing Lactobacillus casei ATCC334 is not significantly different from that of the control group, and is only increased by 14.3%, indicating that Lactobacillus casei ATCC334 does not have the effect of further promoting the reduction of lead ions in intestinal contents by solid beverages. However, the removal effect of lead ions in intestinal contents by solid beverages containing fermented mucus lactobacillus B44 is significantly higher than that of the blank group, the control group and the solid beverage containing the same number of live bacteria, among which it is increased by 46.7% compared with the ATCC334 group, indicating that fermented mucus lactobacillus B44 itself has a certain adsorption effect on lead ions, and its combined effect with solid beverages further promotes the reduction of lead ions in the intestinal environment.

[0088] Effect Example 3: Alleviating effect of solid beverage on lead acetate-induced blood lead in SD rats

[0089] The experimental data are expressed as mean ± standard deviation ( ), where n represents the number of valid samples. SPSS 21.0 software was used to analyze the data. One-way ANOVA statistical method was used for significance test when the data were in accordance with normal distribution, and KW statistical method was used for significance test when the data were not in accordance with normal distribution.

[0090] Weigh 0.25 g of the fermented mucus Lactobacillus solid beverage A prepared according to Example 1, add 10 mL of sterile water, dissolve evenly, and prepare a concentration of 1.0×10 9 CFU / mL bacterial suspension. Thirty-six healthy male SD rats weighing 150-200g, SPF grade, were randomly divided into 3 groups, each containing 12 SD rats, including a negative control group, a lead exposure model group, and a fermented mucus lactobacillus B44 intervention group. According to the grouping, the blank group rats drank blank drinking water normally; the model group and B44 group rats drank an aqueous solution containing 400 mg / L lead ions (accurately weighed 736.5 mg of PbAc2·3H2O, added sterile water to dissolve, fixed to 1L, and mixed). The B44 group rats were fed with 5×10 9 CFU / kg (BW) were gavaged with fermented mucus lactobacillus solid beverage A once a day for 42 consecutive days; the negative control group and model group rats were gavaged with the same dose of unsterilized skim milk once a day for 42 consecutive days. 24 hours after the last administration of the experiment, blood was collected from the orbit (water was not allowed 12 hours before eating) and stored in anticoagulant tubes at -20°C. The blood was used to detect blood lead concentration.

[0091] The blood samples were tested in accordance with the requirements of the national standard "Determination of Lead in Blood Part 2: Inductively Coupled Plasma Mass Spectrometry" (GBZ / T 316.2-2018). The specific steps are as follows: First, place 10 ml of 1% Triton X-100 solution in 1L nitric acid solution (0.5%) to prepare a diluent. Subsequently, take out the refrigerated blood sample and return it to room temperature. After shaking thoroughly, transfer 100 μl of blood sample into a 15 mL polyethylene centrifuge tube, add 10 mL of 0.01% TritonX-100 in 0.5% HNO3 diluent, shake and then use ICP-MS instrument (PerkinElmer NexION 2000) for determination. The specific results are as follows:

[0092] Table 8 Lead concentration in whole blood of SD rats in each group. n=12)

[0093]

[0094] ***, compared with the blank group, P < 0.001; ###, compared with the model group, P < 0.001.

[0095] The results showed that after 6 weeks of the experiment, the lead concentration in the whole blood of SD rats in the modeling group was higher than that in the blank group (P < 0.001). After intervention with fermented Lactobacillus mucus solid beverage, the lead concentration in the whole blood of SD rats was significantly lower than that in the model group (P < 0.001), and the decrease rate reached 49.7%.

[0096] Effect Example 4: Alleviating effect of solid beverage on lead acetate-induced lead accumulation in liver and kidney of SD rats

[0097] The experimental data are expressed as mean ± standard deviation. Indicates, where n represents the number of valid samples. SPSS 21.0 software was used to analyze the data. One-way ANOVA statistical method was used for significance test when the data were in accordance with normal distribution; KW statistical method was used for significance test when the data were not in accordance with normal distribution.

[0098] Weigh 0.25g of the fermented mucus lactobacillus solid beverage A prepared according to Example 1, add 10mL of sterile water, dissolve evenly, and prepare a bacterial suspension with a concentration of 1.0×109CFU / mL. Take 36 healthy male SD rats weighing 150-200g, SPF grade, and randomly divide them into 3 groups, each containing 12 SD rats, including a negative control group, a lead exposure model group, and a fermented mucus lactobacillus B44 intervention group. According to the grouping, the blank group rats drink blank drinking water normally; the model group and the B44 group rats drink a 400mg / L lead ion aqueous solution (accurately weigh 736.5mg of PbAc2·3H2O, add sterile water to dissolve, set to 1L, and mix). The B44 group rats were gavaged with fermented mucus lactobacillus solid beverage A at a dose of 5×109CFU / kg (BW), once a day, for 42 consecutive days; the negative control group and the model group rats were gavaged with the same dose of uninoculated skim milk, once a day, for 42 consecutive days. 24 h after the last administration of the experiment, all rats were killed and liver and kidney tissue samples were taken.

[0099] According to the animal tissue digestion method described in the reference Liang Yanqiu et al. "ICP-MS Determination of Lead, Cadmium and Arsenic in Animal Liver", the tissue was first rinsed with deionized water, dried in an oven at 60°C, and then ground into powder. After nitric acid digestion, the lead content in the tissue was determined using an ICP-MS instrument (PerkinElmer NexION 2000). The specific results are as follows:

[0100] Table 9 Lead content in liver and kidney tissues of SD rats in each group ( n=12)

[0101]

[0102]

[0103] ***, compared with the blank group, P < 0.001; **, compared with the blank group, P < 0.01; ##, compared with the model group, P < 0.01; #, compared with the model group, P < 0.05.

[0104] The results showed that after 6 weeks of the experiment, the lead concentrations in the liver and kidneys of the modeling group were higher than those in the blank group (P < 0.001). After intervention with fermented mucus lactobacillus solid beverage, the lead content in the liver tissue of SD rats was significantly lower than that in the model group (P < 0.01), and the lead concentration in the kidney tissue was significantly lower than that in the model group (P < 0.05). The lead content in the liver and kidney decreased by 44.3% and 24.3% respectively compared with the model group. In addition, data comparison showed that the lead content in the kidney of SD rats was higher than that in the liver. This shows that the intervention with fermented mucus lactobacillus solid beverage has a significant protective effect on the liver and kidney of lead-poisoned SD rats.

[0105] Effect Example 5: Solid beverage promotes the discharge of lead in feces of SD rats induced by lead acetate

[0106] The experimental data are expressed as mean ± standard deviation. Indicates, where n represents the number of valid samples. SPSS 21.0 software was used to analyze the data. One-way ANOVA statistical method was used for significance test when the data were in accordance with normal distribution; KW statistical method was used for significance test when the data were not in accordance with normal distribution.

[0107] Weigh 0.25g of the fermented mucus lactobacillus solid beverage A prepared according to Example 1, add 10mL of sterile water, dissolve evenly, and prepare a bacterial suspension with a concentration of 1.0×109CFU / mL. Take 36 healthy male SD rats weighing 150-200g, SPF grade, and randomly divide them into 3 groups, each containing 12 SD rats, including a negative control group, a lead exposure model group, and a fermented mucus lactobacillus B44 intervention group. According to the grouping, the blank group rats drink blank drinking water normally; the model group and the B44 group rats drink a 400mg / L lead ion aqueous solution (accurately weigh 736.5mg of PbAc2·3H2O, add sterile water to dissolve, set to 1L, and mix). The B44 group rats were gavaged with fermented mucus lactobacillus solid beverage A at a dose of 5×109CFU / kg (BW), once a day, for 42 consecutive days; the negative control group and the model group rats were gavaged with the same dose of uninoculated skim milk, once a day, for 42 consecutive days. 24 h after the last administration of the experiment, all rats were killed and feces samples were collected from the rectum of the rats.

[0108] According to the animal tissue digestion method described in the reference Liang Yanqiu et al. "ICP-MS Determination of Lead, Cadmium and Arsenic in Animal Liver", the tissue was dried in an oven at 60°C, then ground into powder, and then digested with nitric acid and the lead content in the feces was determined using an ICP-MS instrument (PerkinElmer NexION 2000). The specific results are as follows:

[0109] Table 9 Lead content in feces of SD rats in each group ( n=12)

[0110]

[0111] ***, compared with the blank group, P<0.001.

[0112] The results showed that after 6 weeks of the experiment, the lead content in the feces of the modeling group rats was significantly higher than that of the blank group (P < 0.001), and the lead content in the feces of the SD rats after intervention with the fermented mucus lactobacillus solid beverage was higher than that of the model group (P = 0.157), but there was no significant difference. This shows that the intervention with the fermented mucus lactobacillus solid beverage has the effect of promoting fecal lead excretion, which is consistent with the lead reduction effect in the supernatant of intestinal contents in vitro.

[0113] Effect Example 6: Alleviating effect of solid beverage on lead acetate-induced oxidative damage indexes in SD rat kidneys

[0114] The experimental data are expressed as mean ± standard deviation. Indicates, where n represents the number of valid samples. SPSS21.0 software was used to analyze the data. One-way ANOVA statistical method was used for significance test when the data were in accordance with normal distribution, and KW statistical method was used for significance test when the data were not in accordance with normal distribution.

[0115] Weigh 0.25g of the fermented mucus lactobacillus solid beverage A prepared according to Example 1, add 10mL of sterile water, dissolve evenly, and prepare a bacterial suspension with a concentration of 1.0×109CFU / mL. Take 36 healthy male SD rats weighing 150-200g, SPF grade, and randomly divide them into 3 groups, each containing 12 SD rats, including a negative control group, a lead exposure model group, and a fermented mucus lactobacillus B44 intervention group. According to the grouping, the blank group rats drink blank drinking water normally; the model group and the B44 group rats drink a 400mg / L lead ion aqueous solution (accurately weigh 736.5mg of PbAc2·3H2O, add sterile water to dissolve, set to 1L, and mix). The B44 group rats were gavaged with fermented mucus lactobacillus solid beverage A at a dose of 5×109CFU / kg (BW), once a day, for 42 consecutive days; the negative control group and the model group rats were gavaged with the same dose of uninoculated skim milk, once a day, for 42 consecutive days. 24 h after the last administration of the experiment, all rats were killed and kidney tissue samples were taken.

[0116] According to the steps of testing the antioxidant indexes of animal tissue samples described in the reference Wu Chao et al. "Effects of Bamboo Leaf Flavonoids on Liver Damage, Antioxidant Function and Related Gene Expression in Diquat Stress Rats", about 0.2g of kidney sample was taken and stored in liquid nitrogen, and cold saline was used as the homogenization medium to homogenize in an ice water bath at a ratio of 1:9 (mass / volume) using an electric homogenizer until there were no visible tissue particles in the solution. The supernatant was collected after centrifugation at 4°C and 4500×g for 10 minutes. The MDA (malondialdehyde), GSH (glutathione), GSH-PX (glutathione peroxidase), SOD (superoxide dismutase) and CAT (catalase) indicators in the kidney were measured. The relevant indicators were detected using the kit purchased from Nanjing Jiancheng Bioengineering Research Institute, and the operation was carried out strictly in accordance with the instructions of the kit. The specific results are as follows Figure 1 :

[0117] The results showed that after 6 weeks of the experiment, compared with the normal group, the GSH-PX and CAT enzyme activities in the kidneys of the model group rats were significantly reduced, the GSH content was significantly reduced, the MDA content was significantly increased, there was a trend to reduce SOD enzyme activity, and the antioxidant capacity of the kidney was impaired; intervention with fermented mucus Lactobacillus solid beverage can significantly increase the SOD, GSH-PX enzyme activity and GSH content of the model rats' kidneys, and significantly reduce the MDA content, and there is a trend to increase CAT enzyme activity, indicating that the use of fermented mucus Lactobacillus solid beverage can effectively alleviate the renal oxidative damage caused by lead in the body.

[0118] The above examples are for the purpose of illustrating the embodiments disclosed by the present invention and are not to be construed as limiting the present invention. In addition, the various modifications listed herein and the variations of the methods in the invention are obvious to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been specifically described in conjunction with various specific preferred embodiments of the present invention, it should be understood that the present invention should not be limited to these specific embodiments. In fact, various modifications as described above that are obvious to those skilled in the art to obtain the invention should be included within the scope of the present invention.

Claims

1. A solid beverage capable of reducing lead ion emission, characterized in that: The solid beverage comprises Limosilactobacillus fermentum B44 or its active substance and skimmed milk powder, and the preservation number of Limosilactobacillus fermentum B44 is CGMCC No.17321.

2. The solid beverage according to claim 1, characterized in that The active substance is deposited with CGMCC No. The fermentation product of Limosilactobacillus fermentum B44 of 17321; preferably, the fermentation product is selected from the fermentation broth containing bacteria, the fermentation supernatant after removing the bacteria, or the active substance extracted from the fermentation broth.

3. The solid beverage according to claim 1, characterized in that Based on the quality of the solid beverage, the viable count of Lactobacillus fermentum B44 in the solid beverage is 5×10 10 -2x10 11 CFU / g.

4. The solid beverage according to claim 1, characterized in that The solid beverage also contains one or more of water-soluble dietary fiber, sweeteners, vitamins, minerals, edible flavors, edible colorings, and preservatives.

5. The solid beverage according to claim 1, characterized in that: Based on the total weight of the solid beverage, the mass percentage of the fermented mucus lactobacillus B44 or its active substance in the solid beverage is 0.1-50%.

6. Use of the solid beverage according to any one of claims 1 to 5 in the preparation of products for reducing lead toxicity.

7. The use according to claim 6, characterized in that: The lead toxicity reduction product also has any one or more of the following functions: 1) Reduce liver MDA content; 2) Increase CAT enzyme activity; 3) Increase GSH content.

8. The method for preparing a solid beverage according to any one of claims 1 to 5, characterized in that: The preparation method comprises the following steps: adding fermented mucus lactobacillus (Limosilactobacillus fermentum) B44 into skim milk and mixing evenly, freeze-drying to obtain B44 bacterial agent, and then adding sterile skim powder to dilute to obtain a solid beverage.

9. The preparation method according to claim 8, characterized in that: The preparation method comprises the following features: The total weight of the solid beverage is taken as the basis, and the viable count of fermented mucus lactobacillus B44 is 5×10 10 -2x10 11 CFU / g.

10. The preparation method according to claim 8, characterized in that: The preparation method comprises one or more of the following features: I) The skim milk comprises skim milk powder and water, wherein the mass percentage of the skim milk powder to the total mass of the skim milk is 5-15% based on the mass of the skim milk; and / or the mass percentage of water to the total mass of the skim milk is 85-95%. II) Based on the total weight of the solid beverage, the fermented Lactobacillus mucilaginosus B44 or its active substance accounts for 0.1-50% of the solid beverage by weight.

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