CN115024356A - Composite biological preservative special for chilled mutton - Google Patents
Composite biological preservative special for chilled mutton Download PDFInfo
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- CN115024356A CN115024356A CN202210515061.6A CN202210515061A CN115024356A CN 115024356 A CN115024356 A CN 115024356A CN 202210515061 A CN202210515061 A CN 202210515061A CN 115024356 A CN115024356 A CN 115024356A
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
- A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
- A23B4/20—Organic compounds; Microorganisms; Enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
Abstract
The invention discloses a composite biological preservative special for chilled mutton, which comprises the following components: the composite biological preservative is obtained by compounding chitosan, tea polyphenol, grape seed extract and ultrapure water, and the physical and chemical indexes (TVB-N, TBARS and pH), the microbial indexes (high-throughput sequencing and total bacterial count) and the sensory indexes (chromaticity and sensory evaluation) of the mutton stored for 12 days at 4 ℃ after the mutton is treated by the composite biological preservative are analyzed, so that the composite biological preservative can effectively inhibit main spoilage bacteria of the mutton in the cold storage process, and has good bacteriostatic action and fresh-keeping effect.
Description
Technical Field
The invention belongs to the technical field of food preservatives, and particularly relates to a composite biological preservative specially used for chilled mutton.
Background
Mutton has the characteristics of delicious taste, good palatability and rich nutrition, but the shelf life of cold fresh mutton is short, the mutton is easy to deteriorate in the storage process, and for mutton, rich nutrient substances can promote the growth of microorganisms, and meanwhile, the metabolic activity of the mutton can cause the deterioration of the mutton so as to influence the shelf life of the mutton. The deterioration of the quality of the cold fresh meat can be summarized into deterioration and putrefaction caused by the structural change of the cold fresh meat. The main reasons include oxidation, protein autolysis, fatty rancidity and spoilage microorganisms.
The mutton has good palatability and rich nutrition, and has protein content of 18-20% and rich fatty acid, vitamins and minerals. After slaughter, fresh meat is usually stored in refrigeration for a period of time before it is sold and consumed, during which the abundant nutrients in the mutton promote the growth of microorganisms, which colonies of microorganisms (especially spoilage bacteria) grow on their surface first, resulting in spoilage of the meat. The main spoilage bacteria in mutton include pseudomonas (pseudomonas), bacillus (Bacillales) and Flavobacteriales (Flavobacteriales).
At present, preservatives (chemical preservatives and biological preservatives) and physical methods (low-temperature preservation, high-pressure treatment, irradiation, modified atmosphere packaging and the like) are commonly used for preserving the chilled fresh meat, but the physical method for preserving the chilled fresh meat needs large equipment scale, large capital investment and large occupied space. When the chemical preservative is used, the dosage of the chemical preservative exceeds a certain range, and adverse effects such as hepatotoxicity, gastrointestinal diseases, anaphylactic reaction, carcinogenesis or teratogenesis can be caused on the human health. On the contrary, the biological preservative has the advantages of high safety and good bacteriostatic effect, and the application of the biological preservative in the food industry is recently a popular research field. Meanwhile, the antibacterial effect of the composite biological preservative is better than that of a single biological preservative. For mutton, the combination of packaging and refrigeration is the common means for keeping mutton fresh, and among them, air packaging (such as plastic trays wrapped with polyethylene), vacuum packaging and modified atmosphere packaging are the most common types of packaging used for cold fresh mutton. Vacuum and modified atmosphere packaging can significantly delay the growth rate of microorganisms, especially aerobic microorganisms such as Pseudomonas (Pseudomonas) and Acinetobacter (Acinetobacter), in frozen mutton compared to air packaging, thereby extending the shelf life of the mutton. However, such physical preservation methods can only delay the growth of putrefying bacteria, and after the frozen mutton is thawed, the meat quality becomes worse, and the eating quality is influenced. Therefore, the biological preservative with high safety and good bacteriostatic effect has good prospect when being applied to the preservation process of the cold fresh mutton.
Disclosure of Invention
Aiming at the problems, the invention provides the composite biological preservative specially used for the cold fresh mutton, which has good bacteriostatic action and fresh-keeping effect on the cold fresh mutton.
In order to achieve the purpose, the invention adopts the following technical scheme:
a composite biological preservative specially used for chilled mutton comprises the following components in parts by weight:
5-25g of chitosan, 0.125-0.5g of tea polyphenol, 0.125-0.5g of grape seed extract and 1000mL of ultrapure water.
Further, the preservative comprises the following components in parts by weight:
10g of chitosan, 0.5g of tea polyphenol, 0.5g of grape seed extract and 1000mL of ultrapure water.
Further, the preparation method of the preservative comprises the following steps: dissolving 5-25g of chitosan, 0.125-0.5g of tea polyphenol and 0.125-0.5g of grape seed extract in 1000mL of ultrapure water, and uniformly mixing to obtain the composite biological preservative.
By adopting the scheme, the invention has the following advantages:
1. according to the invention, chitosan, tea polyphenol, grape seed extract and water are selected in a targeted manner according to specific spoilage bacteria in the cold fresh mutton to be compounded into a green pollution-free biological preservative, and a high-throughput sequencing result shows that the composite biological preservative can effectively inhibit main spoilage bacteria including pseudomonas, bacillus and flavobacterium in the refrigerating process of the mutton, and has obvious bacteriostatic action and fresh-keeping effect on the cold fresh mutton.
2. The biological preservative is simple to prepare, mild in condition, low in cost, healthy, nontoxic and harmless in raw materials, and has great popularization and application values in the technical field of meat preservation.
3. The biological preservative can prolong the shelf life of mutton by about 3 days, and the oxidation phenomenon of lipid in the mutton is obviously reduced.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Drawings
FIG. 1 shows the change of total number of colonies during storage of mutton at 4 ℃.
Figure 2 is the change in TBARS during storage of mutton at 4 ℃.
Figure 3 is the change of volatile basic nitrogen during the storage of mutton at 4 ℃.
Figure 4 is the sensory index of mutton during storage at 4 ℃.
Figure 5 is the change in pH during storage of mutton at 4 ℃.
FIG. 6 shows the result of high-throughput sequencing of mutton during storage at 4 ℃.
Detailed Description
The present invention will now be described in detail with reference to the drawings and examples, which are not intended to limit the invention in any way, except as specifically stated, reagents, methods and apparatus are conventional in the art.
Example 1: preparation of composite biological preservative specially used for chilled mutton
Dissolving 0.5g of tea polyphenol and 0.5g of grape seed extract in 100mL of sterile ultrapure water, preparing a mixed solution of the tea polyphenol and the grape seed extract, and dissolving 10g of chitosan in the mixed solution to finally obtain the composite biological preservative solution.
Example 2: application of composite biological preservative
1. Experimental methods
1) Pretreatment of mutton
The long dorsal muscles of three small-tail Han sheep within 24 hours after slaughter are purchased from a comprehensive wholesale market in Beibei region of Chongqing, China, and are transported to a laboratory within 30min by using a polyethylene ice bag. The surrounding fat and connective tissue were removed and then divided into cubes of 2X 1 cm. Mutton pieces were randomly selected and divided into two treatment groups.
2) Concentration screening of antistaling agent
Dissolving 0.5g of tea polyphenol and 0.5g of grape seed extract in 100mL of sterile ultrapure water, and preparing into a mixed solution of tea polyphenol and grape seed extract with a total phenol content of 2.5 mg/mL. Then the mixed solution is diluted until the total polyphenol content is 1.2mg/mL and 0.6mg/mL respectively. Dissolving 1g of chitosan into the three mixed solutions to finally obtain three composite biological preservative solutions with different concentrations, as shown in table 1;
TABLE 1 compounding of composite biological antistaling agent solutions with different concentrations
Soaking the cut mutton pieces in the composite biological antistaling agent solution for 30s, taking out, draining, placing into a tray, and sealing with PE (polyethylene) preservative film (25,000 cm) 3 /(m 2 -24h-atm)) and stored at 4 ℃. And (3) taking the total number of the colonies of the mutton as reference, performing single-factor experiments on days 3 and 6, and determining the optimal concentration of the preservative.
2. Results of the experiment
In the single-factor experiment, mutton is treated by using three biological preservatives with different concentrations in the table 1 and then refrigerated at 4 ℃, the colony count of the mutton is measured on the 3 rd day and the 6 th day, according to the single-factor experiment result, the colony count of the control group is the highest, and the numerical values on the 3 rd day and the 6 th day are 4.30 log and 4.93log (CFU/mL) respectively. The high concentration group (10mg/mL chitosan + tea polyphenols with a total phenol content of 2.5mg/mL and grape seed extract) had the lowest total number of colonies at day3 and day 6, 3.85 and 4.15log (CFU/mL), respectively. Therefore, the combination of chitosan (10mg/mL), tea polyphenol and grape seed extract (2.5mg/mL) has good bacteriostatic effect on mutton.
Example 3: physical and chemical index analysis of mutton
1. Experimental Material
Mutton treated by the biological preservative of the high-concentration group (10mg/mL of chitosan, tea polyphenol with the total phenol content of 2.5mg/mL and grape seed extract) in example 2 was taken as a chitosan-tea polyphenol-grape seed extract (CTG) group, and untreated mutton was taken as a blank control group.
2. Experimental methods
1) Determination of pH
Before measuring the pH, the pH meter was calibrated, 5g of treated and untreated mutton were weighed, respectively, minced with a meat mincer, 50mL of ultrapure water was added, and the pH of the homogenate of the meat sample was measured with the pH meter, and 3 sets were made in total.
2) Determination of TBARS
Respectively taking 10g of treated mutton and 10g of untreated mutton, mincing the mutton with a meat mincer, adding 50mL of mixed solution of 7.5% trichloroacetic acid and 0.1% EDTA, shaking for 30min, filtering for 2 times with double-layer filter paper, taking 5mL of supernatant, adding 5mL of TBA (tunnel boring machine) of 0.02mol/L, preserving heat for 40min in a boiling water bath (100 ℃), taking out, cooling for 1h, centrifuging for 5min at 1600r/min, adding 5mL of chloroform into the supernatant, shaking uniformly, standing for layering, taking the supernatant, respectively measuring absorbance at 532nm and 600nm, and making 3 groups of parallel. The TBARS value is calculated according to equation (1).
Wherein A is 532 、A 600 Absorbance measured at wavelength of 532nm and 600nm, respectively; m: sample mass (g).
3) Determination of volatile basic Nitrogen (TVB-N)
Using an automatic Kjeldahl method, 10g of treated mutton and 10g of untreated mutton are respectively taken, minced by a meat grinder, added with 75mL of ultrapure water, shaken, soaked for 30min and filtered. First, a reagent blank measurement was performed, and 1g of magnesium oxide was added to the distillation tube to obtain a blank value. The kjeldahl apparatus settings are as follows: adding 0mL of alkali; 10mL of dilution water; the addition of the boric acid receiver (20g/L) was set at 25 mL; the sample weight was 0 g; the concentration of standard acid is 0.1 mol/L; distillation time 7min and running instrument.
Changing the dilution water to 0mL during formal detection; blank volume is measured blank value; the sample weight was changed to 10g, and the remaining parameters were unchanged. Taking 10mL of filtrate in a distillation tube, adding 1g of magnesium oxide, titrating by using 0.1000mol/L hydrochloric acid standard solution, adding 16.7mL of methyl red ethanol solution and 1g/L of bromocresol green ethanol solution according to the ratio of 1: 5 in a 1000mL, 20g/L boric acid tank, 3 sets were made in parallel. Based on the data displayed on the screen, TVB-N is calculated according to the formula (2)
Wherein V is the volume (mL) of the hydrochloric acid standard titration solution consumed by the test solution; c, the concentration (mol/L) of the hydrochloric acid standard titration solution; m: sample volume (mL).
3. Results of the experiment
1) Measurement of pH: the pH value is an important index for measuring the quality of fresh meat and is related to protein degradation and microbial growth;
as shown in FIG. 5, the pH values of both groups increased with the storage time, and the pH values of both groups tended to increase in general due to decomposition of proteins to produce free amino acids, resulting in NH 3 And the formation of basic reactive compounds such as amines, which manifest as an increase in pH. Furthermore, there was no significant difference (p) between the two groups on each day>0.05), and by day12, the control group and CTG group had pH 8.5 and 8.26, respectively, and finally the control group had slightly higher pH than the CTG group. As chitosan has effective antibacterial activity against various microorganisms (including fungi, yeast and bacteria), pH change can be reduced, and meat spoilage can be inhibited. Therefore, the mutton quality of the CTG group is finally shown to be superior to that of the control group in pH index.
2) Determination of TBARS: TBARS is a good method for detecting lipid peroxidation, and can be used for evaluating the degree of lipid oxidation in mutton, namely the higher the TBARS value is, the higher the degree of lipid oxidation is;
as shown in fig. 2, the TBARS values of the CTG group and the control group Day0 were 0.08 and 0.09mg/kg, respectively, and the trend of the TBARS increase after Day3 in the control group was clear, indicating that the fat oxidation was significant. The difference of TBARS values in the two groups during storage is obvious because the added preservative contains tea polyphenol and polyphenol components in grape seed extract, which have good antioxidant effect, namely the formation of free radicals is inhibited in the initial stage, or the TBARS values in the CTG group are not changed greatly and have obvious difference (p is less than 0.05) from the control group because the added preservative is used as an electron donor to interrupt the proliferation of free radical chain reaction. At Day12, the CTG group was 0.21mg/kg, while the control group reached 2.3mg/kg, which was 10 times the CTG group. The CTG group increased only 0.13mg/kg at the end of storage (Day12) compared to the beginning of storage (Day 0). In conclusion, the addition of tea polyphenol and grape seed extract has good inhibition effect on fat oxidation.
3) Determination of volatile basic nitrogen (TVB-N): the TVB-N is a main component after the protein is decomposed, and the protein is decomposed into nitrogen-containing compounds due to the decomposition effect of enzymes and bacteria in the storage process of the mutton, so that the content of the TVB-N in the mutton can be used for evaluating the freshness of the mutton;
as shown in figure 3, the TVB-N content of two groups of TVB-N is close to 8mg/100g at Day0 in the change condition of the TVB-N in the mutton refrigeration process, and the result shows that the mutton selected in the experiment is fresh and the freshness of the two groups is consistent. The control group increased from 8.13mg/100g to 10.88mg/100g three days before storage, and the change amplitude was larger compared with the CTG group, while the content change of the CTG group was not significantly different (p > 0.05). In addition, both TVB-N values started to have a greater tendency to increase after 9 days of storage. Namely, the TVB-N content is increased along with the increase of the storage time, the national food safety standard (GB2707-2016) stipulates that the TVB-N value of meat does not exceed 15mg/100g, and the TVB-N content of a control group and the TVB-N content of a CTG group are 34.51 and 26.86mg/100g respectively at Day12, and both exceed the upper limit of the standard. From the graph, it can be inferred that the control group may exceed the upper limit on day 10 and the CTG group exceeds the upper limit on day 11.
Example 4: sensory index analysis of mutton
1. Experimental Material
Mutton treated by the biological preservative of the high-concentration group (10mg/mL of chitosan, tea polyphenol with the total phenol content of 2.5mg/mL and grape seed extract) in example 2 was taken as a chitosan-tea polyphenol-grape seed extract (CTG) group, and untreated mutton was taken as a blank control group.
2. Experimental methods
1) Determination of the color
Placing PE preservative film on a table top, respectively taking 5g of treated mutton and untreated mutton, placing the mutton and the untreated mutton on the PE preservative film, and measuring the L of the mutton and the untreated mutton by using a color difference meter * (lightness), a * (degree of Red Green) and b * (yellow-blue) values, 3 replicates were made.
2) Sensory evaluation
A sensory panel consisting of 10 evaluators (6 women and 4 men, 22-25 years old) received specialized training for raw meat, chilled meat (3 days) and spoiled meat evaluations. During the training phase, three meat samples were evaluated daily for two months. The team members then evaluated the samples daily without knowing the storage period. After completion of this exercise with good performance (about two months), the assessors participated in sensory evaluation. Color, smell and texture are used as detection indexes, and the scores of the three indexes are 5, 4, 3, 2 and 1 which respectively represent 5 grades of good, common, poor and poor. Finally, with the weighted sensory index (sensory index SI) as a reference result, the calculation method is as follows: and when the SI is less than or equal to 2.5, the mutton is considered to be unacceptable.
3. Results of the experiment
1) Chroma: table 2 shows the change in color between the control group and the CTG group within 12 days of storage, which is reflected in different L values * 、a * And b * Of the values, L of the CTG group * Values were initially slightly lower than control and did not change significantly during the first few days of storage (p)>0.05), a marked decline (p) starting after Day 6<0.05), this is probably due to the dark reddish brown color of the solution of tea polyphenols and grape seed extract in the compound preservative, i.e. the color of the preservative can cause the color of mutton to be more serious, and meanwhile, the CTG group a is started * And the color is slightly higher than that of the control group, although mutton is bright red, the red color can be deepened after the preservative is added. And b * Values No significant difference (p) between the two groups>0.05), all exhibited a downward trend. In addition, as can be seen from the data in table 2, the browning of the control group is more significant than that of the CTG group, so that although the preservative initially affects the color of the mutton, the preservative still helps to inhibit the browning of the mutton.
TABLE 2 color change of mutton during storage at 4 deg.C
2) Sensory evaluation: as shown in fig. 4, the sensory index of mutton stored for 12 days in cold storage gradually decreased with the increase of storage days. The sensory index of the control group at Day9 was 2.4, which is in an unacceptable range (SI. ltoreq.2.5), while the sensory index of the CTG group decreased to 2.4 at Day 12. However, compared with the control group, the sensory index of the CTG group at Day0 is 4.2, which is lower than that of the control group, because the color and the smell of the tea polyphenol and the grape seed extract solution in the compound preservative have certain influence on the mutton, the scores of the smell and the color are lower than those of the control group. While the grape seed extract, which is presumed to have a large odor effect, was found to have a sensory index lower than that of the CTG group after Day3, and the deterioration of the control group was more significant.
Example 5: microbiological change analysis of mutton
1. Experimental Material
Mutton treated by the biological preservative of the high-concentration group (10mg/mL of chitosan, tea polyphenol with the total phenol content of 2.5mg/mL and grape seed extract) in example 2 was taken as a chitosan-tea polyphenol-grape seed extract (CTG) group, and untreated mutton was taken as a blank control group.
2. Experimental methods
1) Determination of the Total number of colonies
Taking 5.0g of tryptone; 2.5g yeast extract; 1.0g of glucose and 15.0g of agar were dissolved in a conical flask containing 1000mL of pure water and shaken. Placing the test tube, culture medium, normal saline, beaker, pipette tip into autoclave (121 deg.C, 30min), sterilizing, taking out, and cooling;
respectively taking 5g of treated mutton and untreated mutton, placing the treated mutton and the untreated mutton into a sterile homogenizing bag containing 50mL of 0.85% physiological saline, beating for 1min at a medium speed by using a beating type homogenizer, and preparing into a product 1: sample aliquot of 10. Sucking 1mL of 1: 10 sample homogenizing solution by a 1000-L pipette, injecting the sample homogenizing solution into a sterile test tube containing 9mL of physiological saline, and performing gradient dilution to prepare a mixture of 1: 100. 1: 1000. 1: 10000 sample solution. The cooled medium was poured into a 9cm petri dish plate in a clean bench. Sample solutions of different concentrations, 100. mu.L, were pipetted into petri dish plates using a 200. mu.L pipette, one concentration for each of 2 plates. The total number of colonies of the samples of appropriate dilution was determined by plating method and cultured at 36 ℃. + -. 1 for 48 h. + -. 2, and 3 sets of replicates were made. The total number of the bacterial colonies is calculated according to the formula (3);
wherein N is the number of colonies in the sample; sigma C is the sum of the colony counts of the plate (plate containing the colony count in the proper range); n is a radical of an alkyl radical 1 The number of plates with the first dilution (low dilution multiple); n is 2 The number of plates with the second dilution (high dilution multiple); d: dilution factor (first dilution).
2) High-throughput sequencing based on NovaSeq platform
Respectively taking and purifying bacterial DNA in the treated mutton and untreated mutton on the cold storage days 0, 3, 6, 9 and 12, extracting V3-V4 region of 16S rDNA of a sample through PCR amplification, wherein a primer is 341F (CCTACGGGAGGCAGAG), 806R (GGACTACHVGGGTWTCTAAT), carrying out PCR for 3 times, and adding 2 multiplied by 25 mu L of Taq; Primer-F (10. mu.M) 1. mu.L; Primer-R (10. mu.M) 1. mu.L; 50ng of DNA; the nucleic-free water was added to 50uL, and the procedure was as follows: maintaining at 94 deg.C for 5min, respectively performing 30 cycles at 94 deg.C, 52 deg.C and 72 deg.C for 30s, and maintaining at 72 deg.C for 10 min. The length and concentration of the PCR product fragments were checked by 1% agarose gel electrophoresis, and after comparing the concentrations of the PCR products by using GeneTools Analysis Software (V ersonio 4.03.05.0, SynGene), the required volume of each sample was calculated according to the equal mass principle, and each PCR product was mixed. E.z.n.a was used.
The Gel Extraction Kit (Omega, USA) Gel recovery Kit recovers PCR mixed products, Tris-EDTA buffer solution elutes and recovers target DNA fragments, and finally sequencing is carried out on a NovaSeq platform.
3. Results of the experiment
1) Total number of colonies (TVC): the total number of colonies (TVC) may indicate to some extent whether the food is spoiled;
as shown in FIG. 1, the CTG group and the control group were 2.9 and 3.1log CFU/g, respectively, at Day 0. Compared with a control group, on the 12 th day, the total number of the CTG group colonies is reduced by ten times, which can show that the added preservative has a certain inhibiting effect on the microorganisms in the mutton. Along with the increase of the refrigeration time, the difference between the total number of the colonies of the two groups is gradually increased, the growth rate of the control group is slightly higher than that of the CTG group, the control group reaches 6.4log CFU/g at Day9 and exceeds the national food safety standard (GB/T9959.2-2008), the specified allowable limit value of microorganisms is 6log CFU/g, the CTG group reaches 6.1log CFU/g at Day12 and reaches the limit value of the microorganisms 3 days later than the control group, and the fact that the shelf life of the mutton can be prolonged by about 3 days after the preservative is added by taking the total number of the colonies as a reference is further shown.
2) High-throughput sequencing: by combining and filtering the original sequences of each sample, 1107589 high quality effective sequences were obtained in total, with an effective rate of 93.04% -95.62%, the effective sequences were clustered into 2255 OTUs using a similarity threshold of 97%, and the coverage rate for all samples was at least 99%, indicating that almost all bacteria in the mutton sample could be detected. Sequencing analysis detected 15 orders in total, including Pseudomonas, Bacillus, Flavobacterials, Lactobacillus, Aeromonas Enterobacteriales, Micrococcals, Rhizobialles, Corynebacterium, Betaproteobacteriales, Bacteroidales, Clostridium, Alteromonas, Xanthomonas;
the Rank Absundance curve can intuitively reflect two aspects of sample diversity, namely the Abundance and uniformity of species in a sample, and is shown in FIG. 6a, FIG. 6a is a Rank-Absundance curve graph, the curve of Day0 is the most gentle, which indicates that the microbial strains in the mutton are complex at the beginning (Day0), the types are various, the curve is rapidly and steeply reduced until the curve of Day12 is rapidly reduced along with the increase of storage time, the microbial diversity is gradually reduced, and the slope of a CTG group is larger than that of a control group in the same storage days, so that the addition of the biological preservative has an inhibiting effect on the growth of microorganisms;
as shown in fig. 6b, pseudomonas (37.49%), bacillals (16.81%), lactobacillus (20.66%), and aeromonas (14.60%) were major flora, among which pseudomonas is the major dominant bacterium, which together with the "rank-abondance curve" (fig. 6a) demonstrated that only a small fraction of microorganisms caused the decay of mutton. Except for Day 6, the enrichment of Pseudomonas in the CTG group added with the biological preservative is lower than that of the control group, and the key for keeping mutton fresh is to obtain the inhibition of the growth of Pseudomonas;
in the putrefying bacteria of the cold fresh mutton, pseudomonas accounts for about 30 percent as a dominant bacterial colony, in addition, putrefying bacteria such as enterobacter, lactobacillus, staphylococcus and the like are also commonly used as the dominant bacterial colony in the mutton, bacillus is the second dominant bacteria causing mutton putrefaction in the storage period, the richness reaches 47.74 percent of the peak value of Day3 in the CTG group, and then the richness is reduced till 23.40 percent; the bacillus of the control group generally increases and finally reaches 43.72%, so that the chitosan, the tea polyphenol and the grape seed extract have a good inhibition effect on the growth of the bacillus. Meanwhile, the ratio of Lactobacillus is reduced along with the increase of storage time, the control group is reduced from 20.66 percent to 0.59 percent, the CTG group is reduced from 17.63 percent to 2.29 percent, and the composite preservative of chitosan, tea polyphenol and grape seed extract has the most obvious inhibition effect on Lactobacillus. In addition, the proportion of Flavobacteria increases with the increase of the storage time, so that the inhibition of the growth of Flavobacteria can be predicted to be beneficial to the preservation of mutton, and as shown in FIG. 6b, FIG. 6b shows the relative abundance (%) of the bacterial group at the objective level, the enrichment of the bacteria is obviously increased at Day9, and according to the pH detection result, the pH of a control group and a CTG group is respectively 7.5 and 7.2 at Day9, which just belongs to the optimum growth pH condition of the bacteria, so that the reason of the increase of the enrichment of Flavobacteria can be proved;
the visual analysis and comparison of FIG. 6d shows that FIG. 6d is a cluster analysis graph, the colony differences and similarities between groups, the colony compositions of two groups of samples are similar at Day0, and the groups after the start of storage are different. Meanwhile, until Day 6CTG group and the control group were stored for the same time, the difference was not significant, and there was a significant difference from the two groups after Day 6, which is probably due to the exponential growth of the propagation pattern of the microorganisms, which was significantly shown after Day 6. Most notably, the colony composition of the control group was significantly different from that of the CTG group at Day12, and the above total colony content indicated that the control group without the preservative had been significantly deteriorated at Day 12. In addition, a similar conclusion can be obtained according to fig. 6c, and fig. 6c is a PCoA graph, so that the composite preservative of chitosan, tea polyphenol and grape seed extract has a good bacteriostatic effect. In conclusion, a suitable biological preservative is searched in subsequent researches, the growth of three types of bacteria including pseudomonas, Bacillales and flavobacterium can be more specifically inhibited, and the mutton can be better preserved.
Finally, the description is as follows: although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art will understand that various changes, modifications and substitutions can be made without departing from the spirit and scope of the invention as defined by the appended claims. Any modification, equivalent replacement, or improvement made without departing from the spirit and principle of the present invention shall fall within the protection scope of the present invention.
Claims (3)
1. The composite biological preservative specially used for the chilled mutton is characterized by comprising the following components in parts by weight:
5-25g of chitosan, 0.125-0.5g of tea polyphenol, 0.125-0.5g of grape seed extract and 1000mL of ultrapure water.
2. The compound biological preservative specially used for the chilled mutton according to claim 1, wherein the preservative comprises the following components in parts by weight:
10g of chitosan, 0.5g of tea polyphenol, 0.5g of grape seed extract and 1000mL of ultrapure water.
3. The composite biological preservative specially used for the chilled mutton according to claim 1, wherein the preparation method of the preservative comprises the following steps: dissolving 5-25g of chitosan, 0.125-0.5g of tea polyphenol and 0.125-0.5g of grape seed extract in 1000mL of ultrapure water, and uniformly mixing to obtain the composite biological preservative.
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| CN113475568A (en) * | 2021-06-23 | 2021-10-08 | 福建省水产研究所（福建水产病害防治中心） | Composite preservative, preparation method thereof and fresh-keeping method of pufferfish |
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| JP2001078737A (en) * | 1999-09-10 | 2001-03-27 | Oji Paper Co Ltd | Food preservative |
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