Pasteurized modified milk with lead ion emission reduction function and application of pasteurized modified milk

Abstract

The invention relates to the field of modified milk, in particular to pasteurized modified milk with a lead ion emission reduction function and application of the pasteurized modified milk. The pasteurized modified milk provided by the invention contains lactobacillus mucus fermentation B44, wherein the preservation number of the lactobacillus mucus fermentation B44 is CGMCC (China General Microbiological Culture Collection Center) No.17321. Compared with a control sample and other strains, the lactobacillus mucilaginosus B44 in the pasteurized modified milk provided by the invention can remarkably reduce the lead content in intestinal content supernatant, and results of in-vivo experiments of rats show that the lactobacillus mucilaginosus B44 can remarkably reduce the blood lead concentration, the lead content in livers and the lead content in kidneys of drinking objects, so that the lactobacillus mucilaginosus B44 can be used for preparing the pasteurized modified milk. The discharge of lead in excrement can be increased, and liver oxidative damage caused by lead poisoning can be effectively relieved, so that the method can be applied to research on in-vivo emission reduction of lead.

Description

Technical Field

[0001] The invention relates to the field of food biotechnology, in particular to a pasteurized modulated milk with the function of absorbing lead ions and application thereof. Background Art

[0002] Lead (Pb) is a widespread heavy metal that poses a serious threat to the environment and human health. Unlike organic pollutants, heavy metals such as lead are difficult to decompose or volatilize through natural processes. Their persistence in the environment allows them to accumulate through the food chain, ultimately causing serious impacts on human health. Lead is particularly toxic to children, and it may cause behavioral problems, learning disabilities, and delayed growth and development.

[0003] At present, there are two main methods to reduce the lead content in the body: one is to use lead chelating agents, such as succimer and sodium calcium edetate, which can bind to lead and promote its excretion from the body. However, these drugs may have side effects and are questionable for the safety of sensitive groups such as children and the elderly, so they are not recommended as routine treatment.

[0004] In order to protect the public health, especially children and the elderly, from the threat of lead pollution, it is necessary to actively seek safer and more effective alternative treatments. These new treatments may include nutritional interventions to reduce lead absorption and toxicity by supplementing certain nutrients, or the development of new chelating agents, which aim to reduce side effects and improve safety. Summary of the invention

[0005] In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to provide a pasteurized formula milk with the function of reducing lead ion emission and application thereof, so as to solve the problems in the prior art.

[0006] To achieve the above purpose and other related purposes, the present invention provides a pasteurized modulated milk with the function of reducing lead ion emission. The pasteurized modulated milk contains fermented Lactobacillus mucilaginosus B44, and the preservation number of the fermented Lactobacillus mucilaginosus B44 is CGMCC No.17321.

[0007] Preferably, based on the volume of the pasteurized prepared milk, the number of viable bacteria of fermented Lactobacillus mucilaginosus B44 in the pasteurized prepared milk is 1×10 6 -5x10 9 CFU / ml.

[0008] The present invention also provides a method for preparing the pasteurized modulated milk, which comprises the following steps: adding fermented mucus lactobacillus B44 into pasteurized milk and mixing to obtain the obtained product, wherein the preservation number of fermented mucus lactobacillus B44 is CGMCC No.17321.

[0009] The present invention also provides the use of the pasteurized modulated milk in the preparation of a heavy metal ion adsorption product.

[0010] As described above, the pasteurized modulated milk with the function of adsorbing lead ions and the application thereof of the present invention have the following beneficial effects:

[0011] The present invention provides a Lactobacillus mucilaginosus B44 fermented in pasteurized modulated milk, which can significantly reduce the lead content in the supernatant of intestinal contents compared with a control sample and other strains, and from the results of in vivo experiments on rats, it can be known that the blood lead concentration, the lead content in the liver and the lead content in the kidney of the drinking subject can be significantly reduced, and the discharge of lead in feces can be increased and the liver oxidative damage caused by lead poisoning can be effectively alleviated. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 Shown are the test results of antioxidant indexes in liver of SD rats after intervention with lead poisoning and pasteurized milk prepared by fermentation of Lactobacillus mucilaginosus B44 in the present invention. DETAILED DESCRIPTION

[0013] The invention provides a pasteurized modulated milk with the function of absorbing lead ions. The pasteurized modulated milk contains Limosilactobacillus fermentum B44.

[0014] 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.

[0015] In some specific embodiments, based on the volume of the pasteurized prepared milk, the number of viable bacteria of fermented Lactobacillus mucilaginosus B44 in the pasteurized prepared milk is 1×10 6 -5x10 9 CFU / ml. Specifically, the viable count is 1x10 6 -5x10 6 CFU / ml, 5x10 6 -1x10 7 CFU / ml, 1x10 7 -5x10 7 CFU / ml, 5x10 7 -1x10 8 CFU / ml or 1x10 8 -5x10 9 CFU / ml. Preferably, the viable count is 5x10 8-5x10 9 CFU / ml.

[0016] In some specific embodiments, the pasteurized modulated milk further contains one or more of lipids, proteins, emulsifiers, sweeteners, vitamins, minerals, food flavors, food colorings, preservatives, stabilizers, thickeners or acidity regulators.

[0017] In some embodiments, the lipid is selected from animal fat or vegetable fat. Specifically, the animal fat can be selected from cream; or the vegetable fat is selected from one or more of coconut oil, palm oil, rapeseed oil, olive oil, peanut oil, camellia seed oil or sunflower seed oil.

[0018] In some embodiments, the protein may be selected from whey protein or casein.

[0019] In some specific embodiments, the emulsifier may be a lipophilic emulsifier, or a combination of a lipophilic emulsifier and a hydrophilic emulsifier. Specifically, the lipophilic emulsifier includes one or two of monoglyceride, mono- and diglyceride, and sodium stearate lactate; or, the hydrophilic emulsifier is one or two of sucrose ester and polyglycerol ester.

[0020] In some specific embodiments, the sweetener can be selected from one or more of honey, sucrose, sucralose, fructose, glucose, erythritol, xylitol, corn syrup or fructose syrup.

[0021] 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.

[0022] In some specific embodiments, the mineral may be selected from one or more of sodium salts, potassium salts, magnesium salts, calcium salts or phosphates.

[0023] 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.

[0024] In some specific embodiments, the stabilizer or thickener can be selected from one or more of pectin, sodium carboxymethyl cellulose, xanthan gum, sodium alginate, carrageenan, gellan gum, monoglyceride, sucrose ester or locust bean gum.

[0025] In some embodiments, the acidity regulator can be selected from one or more of citric acid or its salts, malic acid or its salts, gluconic acid or its salts, or tartaric acid or its salts.

[0026] The present invention also provides a method for preparing the pasteurized modulated milk, which comprises the following steps: adding fermented mucus lactobacillus B44 into pasteurized milk and mixing to obtain the obtained product, wherein the preservation number of fermented mucus lactobacillus B44 is CGMCC No.17321.

[0027] In some specific embodiments, based on the volume of the pasteurized prepared milk, the number of viable bacteria of fermented Lactobacillus mucilaginosus B44 in the pasteurized prepared milk is 1×10 6 -1x10 9 CFU / ml. Specifically, the viable count is 1x10 6 -5x10 6 CFU / ml, 5x10 6 -1x10 7 CFU / ml, 1x10 7 -5x10 7 CFU / ml, 5x10 7 -1x10 8 CFU / ml, 1x10 8 -5x10 8 CFU / ml, 5x10 8 -1x10 9 CFU / ml or 1x10 9 -5x10 9 CFU / ml. Preferably, the viable count is 5x10 8 -5x10 9 CFU / ml.

[0028] In some embodiments, the pasteurized milk can be selected from cow's milk, goat's milk or reconstituted milk.

[0029] In some specific embodiments, when the pasteurized milk is cow's milk or goat's milk, the pasteurized milk is obtained by the following steps:

[0030] The raw cow's milk or raw goat's milk is obtained by purifying, homogenizing and sterilizing the raw milk.

[0031] In the present invention, the raw milk is milk received in accordance with the relevant requirements and regulations in GB19301-2010.

[0032] In some specific embodiments, the milk purification step is to cool the temperature of the raw cow's milk or raw goat's milk to 2-6° C. Specifically, the milk purification step is to cool the temperature of the raw cow's milk or raw goat's milk to 2-3° C., 3-4° C., 4-5° C. or 5-6° C.

[0033] In some specific embodiments, the temperature during the homogenization process is 62-64°C and the pressure is 18-25 MPa. Specifically, the temperature during the homogenization process is 62-63°C or 63-64°C; the pressure is 18-20 MPa, 20-22 MPa, 22-23 MPa or 23-25 ​​MPa.

[0034] In some embodiments, the sterilization can be selected from any one of the following steps:

[0035] 1) maintaining at 60-65°C for 25-35 min; preferably, maintaining at 63°C for 29-31 min;

[0036] 2) Maintain at 72°C-75°C or 82°C-85°C for 15-20 seconds;

[0037] 3) Maintain at 135℃-140℃ for 2-5s; maintain at 135℃-140℃ for 3-4s.

[0038] In some specific embodiments, the preparation method further comprises a canning step, wherein the canning step comprises cooling the mixed emulsion to 2-6°C for canning.

[0039] The present invention also provides the use of the pasteurized modulated milk in preparing a heavy metal ion adsorption product.

[0040] In some specific embodiments, the heavy metal ion adsorption product is a product with antioxidant function.

[0041] Furthermore, the antioxidant function includes one or more of the following characteristics:

[0042] 1) Reduce liver MDA content;

[0043] 2) Increase CAT enzyme activity;

[0044] 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.

[0045] The pasteurized modulated milk provided by the present invention has an improvement in the removal effect of lead ions in the supernatant of intestinal contents by ≥28.6% compared with the control group, reduces the blood lead concentration of the lead model SD rats by 50.9% compared with the model group, reduces the lead content of the liver and kidney of the lead model SD rats by 44.4% and 27.1% respectively compared with the control group, and has the effect of promoting the excretion of lead in feces of the lead model SD rats.

[0046] In the present invention, the term "reconstituted milk" generally refers to milk powder (which may be whole milk powder or skim milk powder) mixed with water to form an emulsion having a ratio of water to solids equivalent to that of the original milk.

[0047] 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.

[0048] 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.

[0049] 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.

[0050] Example 1

[0051] 1. Materials and methods

[0052] (a) Preparation of fermented Lactobacillus mucilaginosus B44 bacterial agent

[0053] 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 h. 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 h, and inoculated into MRS liquid medium with an inoculum of 3% (v / v). After culturing again at 37°C for 24 h, the culture was centrifuged at 9,000 rpm and 7°C for 15 minutes, the supernatant was discarded, the bacteria were washed once with sterile water, and then suspended with a small amount of skim milk (10%, v / w), and freeze-dried to obtain the bacterial agent of fermented mucus lactobacillus B44. The concentration of the bacterial agent was determined to be 4.0×10 10 CFU / g.

[0054] (b) Preparation of pasteurized milk

[0055] The raw milk is purified, homogenized and pasteurized in sequence.

[0056] 2. Preparation of pasteurized milk: Fermented Lactobacillus mucilaginosus B44 was added to a final concentration of 4 x 10 9 CFU / mL is added into the pasteurized milk and mixed evenly to obtain the pasteurized prepared milk A of fermented Lactobacillus mucilaginosus of this embodiment.

[0057] Example 2

[0058] 1. Materials and methods

[0059] (a) Preparation of fermented Lactobacillus mucilaginosus B44 bacterial agent: Same as Example 1

[0060] (b) Preparation of pasteurized milk: Same as Example 1

[0061] 2. Preparation of pasteurized milk fermented with Lactobacillus mucilaginosus B44

[0062] The fermented Lactobacillus mucilaginosus B44 was added to a final concentration of 6x10 8 CFU / mL is added into the pasteurized milk and mixed evenly to obtain the pasteurized prepared milk B of fermented Lactobacillus mucilaginosus of this embodiment.

[0063] Example 3

[0064] 1. Materials and methods

[0065] (a) Preparation of fermented Lactobacillus mucilaginosus B44 bacterial agent: Same as Example 1

[0066] (b) Preparation of pasteurized milk: Same as Example 1

[0067] 2. Preparation of pasteurized milk fermented with Lactobacillus mucilaginosus B44

[0068] The fermented Lactobacillus mucilaginosus B44 was added to a final concentration of 8x10 8 CFU / mL is added into the pasteurized milk and mixed evenly to obtain the pasteurized prepared milk C of fermented Lactobacillus mucilaginosus of this embodiment.

[0069] Example 4

[0070] 1. Materials and methods

[0071] (a) Preparation of fermented Lactobacillus mucilaginosus B44 bacterial agent: Same as Example 1

[0072] (b) Preparation of pasteurized milk: Same as Example 1

[0073] 2. Preparation of pasteurized milk fermented with Lactobacillus mucilaginosus B44

[0074] The fermented Lactobacillus mucilaginosus B44 was added to a final concentration of 1x10 9 CFU / mL was added into the pasteurized milk and mixed evenly to obtain the pasteurized prepared milk D of fermented Lactobacillus mucilaginosus of this embodiment.

[0075] Example 5

[0076] 1. Materials and methods

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

[0078] (b) Preparation of pasteurized milk: Same as Example 1

[0079] 2. Preparation of pasteurized milk fermented with Lactobacillus mucilaginosus B44

[0080] The fermented Lactobacillus mucilaginosus B44 was added to a final concentration of 5x10 9 CFU / mL is added into the pasteurized milk and mixed evenly to obtain the pasteurized prepared milk E of fermented Lactobacillus mucilaginosus of this embodiment.

[0081] Effect Example 1: Effect of pasteurized milk fermented with Lactobacillus mucilaginosus on reducing lead ion emission in intestinal contents

[0082] (a) In vitro preparation of intestinal contents

[0083] 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.

[0084] (b) Experiment on reduction of lead ions in intestinal contents

[0085] Take 1 mL of each of the fermented mucus lactobacillus pasteurized milk A to E of Examples 1 to 5 above (the blank group was without any treatment; the pasteurized cow milk without treatment and without bacteria was the control group), and mix them with 19 mL of the intestinal content solution containing lead ions, so that the final lead ion concentration is 50 mg / L. After oscillating at 37°C and 200 rpm for 1 hour, centrifuge at 9,000 rpm and 7°C for 15 minutes, and collect the supernatant solution. Take 5 mL of the above supernatant solution, add 3 mL of concentrated nitric acid (super pure), and digest it in a graphite digester at 130°C for 30 minutes. Rinse the tube twice with deionized water, adjust the volume to 15 mL, and measure it using an inductively coupled plasma optical emission spectrometer (ICP-OES). The specific results are as follows:

[0086] Table 1 Effect of removing lead ions from intestinal contents by fermented mucus lactobacillus milk of Examples 1 to 5

[0087]

[0088] As can be seen from the above table, intestinal contents have a certain adsorption effect on lead ions in water, resulting in a lower lead content in the supernatant solution of the blank group (without any treatment) than in the initially set solution system. This is due to the adsorption of lead by bacteria and organic matter in the intestinal contents; and although the control group (unbacteria-free pasteurized milk) itself has a certain reduction effect on lead ions in the intestine, when fermented Lactobacillus mucus B44 is added to the pasteurized milk, the removal effect of lead is further improved. Specifically, the removal effect of lead ions in the supernatant of intestinal contents by fermented Lactobacillus mucus pasteurized formula milk is ≥35.9% higher than that of the blank group, and ≥28.6% higher than that of the control group.

[0089] Effect Example 2 Refrigeration Experiment of Fermented Lactobacillus Mucilaginosus Pasteurized Modified Milk

[0090] The fermented Lactobacillus mucilaginosus pasteurized formula milks A to E of Examples 1 to 5 were stored under refrigeration (7° C.) and taken out at 0, 3 and 7 days. The lead removal rate of each sample was measured by referring to the method described in Effect Example 1. The results are shown in the following table.

[0091] Table 2 The removal effect of lead ions in intestinal contents by fermented Lactobacillus mucilaginosus milk AE during storage

[0092]

[0093] As can be seen from the above table, the adsorption rate of lead by the pasteurized fermented Lactobacillus mucilaginosus prepared by the present invention remains stable after being stored under refrigeration (7° C.) for 1 week.

[0094] Comparative Example 1

[0095] With reference to the method described in Example 1, the removal effect of lead ions in intestinal contents by pasteurized formula milk 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 operation was as follows.

[0096] 1. Materials and methods

[0097] (a) Preparation of fermented Lactobacillus mucilaginosus B44 bacterial agent: Same as Example 1

[0098] (b) Preparation of Lactobacillus casei ATCC334 bacterial agent

[0099] 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.

[0100] 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, the bacteria were washed once with sterile water, suspended with a small amount of skim milk (10%, v / w), and freeze-dried to obtain the bacterial agent of Lactobacillus casei ATCC334. The concentration of the bacterial agent was determined to be 5.8×10 10 CFU / g.

[0101] Preparation of pasteurized milk: Same as Example 1

[0102] 2. Preparation of pasteurized lactobacillus milk

[0103] The fermented Lactobacillus mucilaginosus B44 and Lactobacillus casei ATCC334 were added to a final concentration of 4×10 9 CFU / mL were added to the pasteurized milk and mixed evenly to obtain pasteurized modulated milk prepared by adding different lactic acid bacteria.

[0104] 3. Comparison of the reduction effect of lead ions in intestinal contents

[0105] The removal effects of the two pasteurized formula milks on lead ions in the intestinal contents were measured by the method described in Reference Effect Example 1. The results are shown in the following table.

[0106] Table 3 Effect of different lactobacillus-modified milk on the removal of lead ions from intestinal contents

[0107] Lactobacillus formula milk blank Comparison B44 ATCC334 Lead ion concentration in supernatant (mg / L) 3.9 3.5 1.8 3.3

[0108] By comparison, it can be seen that the removal effect of lead ions in intestinal contents by pasteurized modulated milk containing Lactobacillus casei ATCC334 is not significantly different from that of the control group, and is only increased by 5.7%, indicating that Lactobacillus casei ATCC334 does not have the effect of promoting the reduction of lead ions in intestinal contents by pasteurized cow's milk. However, the removal effect of lead ions in intestinal contents by pasteurized modulated milk containing fermented mucus lactobacillus B44 is significantly higher than that of the blank group, the control group and the pasteurized modulated milk containing ATCC334 with the same number of live bacteria, among which it is increased by 45.5% compared with the pasteurized modulated milk of ATCC334, indicating that fermented mucus lactobacillus B44 itself has a certain adsorption effect on lead ions in the solution system, and its combined action with cow's milk further promotes the reduction of lead ions in the intestinal environment.

[0109] Effect Example 3: The alleviating effect of pasteurized milk on lead acetate-induced blood lead in SD rats

[0110] 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.

[0111] The pasteurized milk D prepared by fermentation of Lactobacillus mucilaginosus according to Example 4 is pasteurized milk prepared by fermentation of Lactobacillus mucilaginosus B44 with a final concentration of 1x109 CFU / mL. Thirty-six healthy male SD rats weighing 150-200g, SPF grade, were randomly divided into three groups, each containing 12 SD rats, including a negative control group, a lead exposure model group and a fermentation of Lactobacillus mucilaginosus 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 400mg / L lead ions (accurately weigh 736.5mg of PbAc2·3H2O, add sterile water to dissolve, dilute to 1L, and mix well). The rats in the B44 group drank 5×10 9 CFU / kg (BW) were gavaged with pasteurized milk D once a day for 42 consecutive days; the negative control group and model group rats were gavaged with the same dose of unsterilized pasteurized milk once a day for 42 consecutive days. 24 hours after the last administration of the experiment, blood was collected from the eye sockets (water was not allowed 12 hours before eating), stored in anticoagulant tubes at -20°C, and the blood was used to detect blood lead concentration.

[0112] 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:

[0113] Table 8 Lead concentration in whole blood of SD rats in each group. n=12)

[0114]

[0115] ***, compared with the blank group, P < 0.001; ###, compared with the model group, P < 0.001.

[0116] 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 pasteurized milk prepared with fermented Lactobacillus mucosa, the lead concentration in the whole blood of SD rats was significantly lower than that in the model group (P < 0.001), with a decrease rate of 50.9%.

[0117] Effect Example 4: The alleviating effect of pasteurized formula milk on lead acetate-induced lead accumulation in the liver and kidney of SD rats

[0118] 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.

[0119] The fermented Lactobacillus mucilaginosus pasteurized milk D prepared in Example 4, i.e., the fermented Lactobacillus mucilaginosus B44 agent was prepared at a final concentration of 1×10 9CFU / mL pasteurized formula milk. Thirty-six healthy male SD rats weighing 150-200g, SPF grade, were randomly divided into three 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 400mg / L lead ions (accurately weighed 736.5mg 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 pasteurized milk D once a day for 42 consecutive days; the negative control group and model group rats were gavaged with the same dose of unsterilized pasteurized milk once a day for 42 consecutive days. 24 hours after the last administration of the experiment, all rats were killed and liver and kidney tissue samples were taken.

[0120] 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:

[0121] Table 9 Lead content in liver and kidney tissues of SD rats in each group ( n=12)

[0122]

[0123] ***, 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.

[0124] The results showed that after 6 weeks of the experiment, the lead concentrations in the liver and kidney of the modeling group were higher than those in the blank group (P < 0.001). After intervention with fermented Lactobacillus mucus pasteurized milk, 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 44.4% and 27.1% 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 drinking fermented Lactobacillus mucus pasteurized milk has a significant protective effect on the liver and kidney of lead-poisoned SD rats.

[0125] Effect Example 5: Pasteurized milk promotes the discharge of lead in feces of SD rats induced by lead acetate

[0126] 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.

[0127] The pasteurized milk D prepared by fermentation of Lactobacillus mucilaginosus according to Example 4 is pasteurized milk prepared by fermentation of Lactobacillus mucilaginosus B44 with a final concentration of 1x109 CFU / mL. Thirty-six healthy male SD rats weighing 150-200g, SPF grade, were randomly divided into three groups, each containing 12 SD rats, including a negative control group, a lead exposure model group and a fermentation of Lactobacillus mucilaginosus 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 400mg / L lead ions (accurately weigh 736.5mg of PbAc2·3H2O, add sterile water to dissolve, dilute to 1L, and mix well). The rats in the B44 group drank 5×10 9 CFU / kg (BW) were gavaged with pasteurized milk D once a day for 42 consecutive days; the negative control group and model group rats were gavaged with the same dose of unsterilized pasteurized milk once a day for 42 consecutive days. 24 hours after the last administration of the experiment, all rats were killed and fecal samples were taken from the rat rectum.

[0128] 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:

[0129] Table 9 Lead content in feces of SD rats in each group ( n=12)

[0130]

[0131] ***, compared with the blank group, P<0.001.

[0132] 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 fermented mucus lactobacillus pasteurized milk was higher than that of the model group (P = 0.225), but there was no significant difference. This shows that drinking fermented mucus lactobacillus pasteurized milk has a certain effect of promoting fecal lead excretion in the body, which is consistent with the lead reduction effect in the supernatant of intestinal contents in vitro.

[0133] Effect Example 6: The alleviating effect of pasteurized modified milk on the liver oxidative damage indexes of SD rats induced by lead acetate

[0134] 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.

[0135] The pasteurized milk D prepared by fermentation of Lactobacillus mucilaginosus according to Example 4 is pasteurized milk prepared by fermentation of Lactobacillus mucilaginosus B44 with a final concentration of 1x109 CFU / mL. Thirty-six healthy male SD rats weighing 150-200g, SPF grade, were randomly divided into three groups, each containing 12 SD rats, including a negative control group, a lead exposure model group and a fermentation of Lactobacillus mucilaginosus 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 400mg / L lead ions (accurately weigh 736.5mg of PbAc2·3H2O, add sterile water to dissolve, dilute to 1L, and mix well). The rats in the B44 group drank 5×10 9 CFU / kg (BW) were gavaged with pasteurized milk D once a day for 42 consecutive days; the negative control group and model group rats were gavaged with the same dose of unsterilized pasteurized milk once a day for 42 consecutive days. 24 hours after the last administration of the experiment, all rats were killed and liver tissue samples were taken.

[0136] According to the steps of testing liver antioxidant indexes described in the reference Wu Chao et al. "Effects of Bamboo Leaf Flavonoids on Liver Damage, Antioxidant Function and Related Gene Expression in Diquat-Stressed Rats", about 0.2 g of liver 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. MDA (malondialdehyde), GSH (glutathione), GSH-PX (glutathione peroxidase), SOD (superoxide dismutase) and CAT (catalase) indicators in the liver were determined. 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 :

[0137] The results showed that after 6 weeks of the experiment, compared with the normal group, the SOD and CAT enzyme activities in the liver of SD rats in the model group were significantly reduced, the GSH content was significantly reduced, the GSH-PX enzyme activity showed a downward trend, and the MDA content showed an upward trend, indicating that the antioxidant capacity of the liver of lead-poisoned rats was reduced; intervention with fermented mucus lactobacillus pasteurized milk could significantly reduce the MDA content in the liver of rats in the model group, and there was a trend of increasing CAT enzyme activity and GSH content, indicating that drinking fermented mucus lactobacillus B44 pasteurized milk can effectively alleviate the liver oxidative damage caused by lead poisoning.

[0138] 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 pasteurized formula milk having the function of absorbing lead ions, characterized in that: The pasteurized formulated milk contains Limosilactobacillus fermentum B44 and pasteurized milk, and the preservation number of the Limosilactobacillus fermentum B44 is CGMCC No.17321.

2. The pasteurized formulated milk according to claim 1, characterized in that: Based on the volume of pasteurized milk, the number of viable bacteria of Lactobacillus fermentum B44 in the pasteurized milk is 1×10 6 -5x10 9 CFU / ml; preferably, the viable count is 5x10 8 -5x10 9 CFU / ml.

3. The pasteurized formulated milk according to claim 1, characterized in that: The pasteurized modulated milk further contains one or more of lipid, protein, emulsifier, sweetener, vitamin, mineral substance, edible flavor, edible coloring, preservative, stabilizer, thickener or acidity regulator.

4. The method for preparing pasteurized formulated milk according to claim 1 or 2, characterized in that: The preparation method comprises the following steps: adding Limosilactobacillus fermentum B44 into pasteurized milk and mixing to obtain the obtained product, wherein the preservation number of Limosilactobacillus fermentum B44 is CGMCC No.17321.

5. The preparation method according to claim 4, characterized in that: Based on the volume of pasteurized milk, the number of viable bacteria of Lactobacillus fermentum B44 in the pasteurized milk is 1×10 6 -5x10 9 CFU / ml; preferably, the viable count is 5x10 8 -5x10 9 CFU / ml.

6. The preparation method according to claim 4, characterized in that: The pasteurized milk is selected from cow's milk, goat's milk or reconstituted milk.

7. The preparation method according to claim 6, characterized in that, when the pasteurized milk is cow's milk or goat's milk, the pasteurized milk is obtained by the following steps: purifying, homogenizing and sterilizing raw cow's milk or raw goat's milk in sequence.

8. Use of the pasteurized modulated milk according to any one of claims 1 to 4 in the preparation of heavy metal ion adsorption products.

9. The use according to claim 8, characterized in that: The heavy metal ion adsorption product is a product with antioxidant function.

10. The method according to claim 9, characterized in that: The antioxidant function includes one or more of the following characteristics: 1) Reduce liver MDA content; 2) Increase CAT enzyme activity; 3) Increase GSH content.

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