CN117441780A - Method for reducing bacteria of chilled chicken - Google Patents
Method for reducing bacteria of chilled chicken Download PDFInfo
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- CN117441780A CN117441780A CN202311524461.4A CN202311524461A CN117441780A CN 117441780 A CN117441780 A CN 117441780A CN 202311524461 A CN202311524461 A CN 202311524461A CN 117441780 A CN117441780 A CN 117441780A
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- chicken
- sodium hypochlorite
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- bacteria
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- 241000287828 Gallus gallus Species 0.000 title claims abstract description 103
- 241000894006 Bacteria Species 0.000 title claims abstract description 47
- 238000000034 method Methods 0.000 title claims abstract description 27
- 230000001603 reducing effect Effects 0.000 title claims abstract description 16
- 239000005708 Sodium hypochlorite Substances 0.000 claims abstract description 54
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims abstract description 52
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 claims abstract description 39
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 36
- 238000002791 soaking Methods 0.000 claims abstract description 18
- 238000009210 therapy by ultrasound Methods 0.000 claims abstract description 14
- 238000004806 packaging method and process Methods 0.000 claims abstract description 9
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- 230000009467 reduction Effects 0.000 abstract description 32
- 230000002195 synergetic effect Effects 0.000 abstract description 18
- 238000005516 engineering process Methods 0.000 abstract description 6
- 238000011282 treatment Methods 0.000 description 91
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- 239000011780 sodium chloride Substances 0.000 description 2
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- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
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- FRPHFZCDPYBUAU-UHFFFAOYSA-N Bromocresolgreen Chemical compound CC1=C(Br)C(O)=C(Br)C=C1C1(C=2C(=C(Br)C(O)=C(Br)C=2)C)C2=CC=CC=C2S(=O)(=O)O1 FRPHFZCDPYBUAU-UHFFFAOYSA-N 0.000 description 1
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 1
- QDHHCQZDFGDHMP-UHFFFAOYSA-N Chloramine Chemical compound ClN QDHHCQZDFGDHMP-UHFFFAOYSA-N 0.000 description 1
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- 239000011777 magnesium Substances 0.000 description 1
- 239000000395 magnesium oxide Substances 0.000 description 1
- CPLXHLVBOLITMK-UHFFFAOYSA-N magnesium oxide Inorganic materials [Mg]=O CPLXHLVBOLITMK-UHFFFAOYSA-N 0.000 description 1
- AXZKOIWUVFPNLO-UHFFFAOYSA-N magnesium;oxygen(2-) Chemical compound [O-2].[Mg+2] AXZKOIWUVFPNLO-UHFFFAOYSA-N 0.000 description 1
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- CEQFOVLGLXCDCX-WUKNDPDISA-N methyl red Chemical compound C1=CC(N(C)C)=CC=C1\N=N\C1=CC=CC=C1C(O)=O CEQFOVLGLXCDCX-WUKNDPDISA-N 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
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- DUIOPKIIICUYRZ-UHFFFAOYSA-N semicarbazide Chemical compound NNC(N)=O DUIOPKIIICUYRZ-UHFFFAOYSA-N 0.000 description 1
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- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
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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
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/14—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
- A23B7/153—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
- A23B7/157—Inorganic compounds
-
- 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
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/015—Preserving by irradiation or electric treatment without heating effect
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Inorganic Chemistry (AREA)
- Meat, Egg Or Seafood Products (AREA)
Abstract
The invention discloses a method for reducing bacteria of chilled chicken, which comprises the following steps: (1) Soaking the frozen fresh chicken in a low-concentration sodium hypochlorite solution, and performing ultrasonic treatment; (2) And (3) soaking the chicken processed in the step (1) in an ozone water solution, and then draining, packaging in a tray and refrigerating. The invention combines the ozone water bacteria reduction technology and the ultrasonic synergistic sodium hypochlorite bacteria reduction technology, provides a novel, green and more effective bacteria reduction mode, and effectively prolongs the shelf life of the chilled chicken.
Description
Technical Field
The invention relates to the technical field of food processing, in particular to a method for reducing bacteria in chilled chicken.
Background
The frozen fresh chicken is the fresh chicken which is obtained by killing the live chicken which is qualified by quarantine inspection, quickly reducing the carcass temperature to 8 ℃, reducing the carcass temperature to 4 ℃ within 12 hours, and cutting, processing, circulating, storing and distributing the live chicken within the temperature range of 0-4 ℃. The frozen fresh chicken is popular with consumers due to higher nutritive value and delicious taste, the market demand is continuously increased, but rich nutritive substances in the frozen fresh chicken are beneficial to the growth and propagation of microorganisms, the spoilage in the production and sales process causes great resource waste for slaughter enterprises, the chicken quality during the sales period is also negatively influenced, and the frozen fresh chicken is an important problem which is difficult to solve by the food industry. Chicken skin is one of the most serious parts of chicken carcasses, and has the characteristics of easy putrefaction, difficult cleaning and the like. The chicken skin is positioned at an environmental interface, so that the parts of the chicken skin, feathers and the like exposed to the outside are always at a higher microorganism level in the processes of raising, transporting and the like of the broiler chickens. In addition, the chicken skin is directly contacted with pre-cooling water, air, an operation surface and operators in the slaughtering processing link to further pollute the surface of the chicken carcass, so that the chicken skin turns yellow and is sticky, and the chicken skin is spoiled. Meanwhile, after the broiler chicken is slaughtered and roughened, the hair follicles on the chicken skin can be closed in a short time, so that a large amount of microorganisms remain in the hair follicles, and the closed microorganisms are difficult to clean and difficult to act by the degerming agents such as sodium hypochlorite. This will further exacerbate the perishable nature of chicken skin. In the market of China, a large number of high-value chicken products (chicken wings, chicken legs and the like) with skin exist, and the spoilage of chicken skin parts can lead to the overall spoilage. This is a problem that many people currently ignore. Therefore, the cleaning and bacteria reduction degree of the chicken skin has important significance for maintaining the shelf life of the fresh chicken
At present, the broiler industry mainly adopts a sodium hypochlorite soaking method to disinfect broiler chickens. Sodium hypochlorite is used as an oxidizing bactericide, and is favored by the food industry due to its broad bactericidal spectrum and low price. However, sodium hypochlorite is a chemical microbial reducing agent, and a high addition amount of sodium hypochlorite can react with organic matters to generate byproducts such as trihalomethane, chloramine, halogenated ketone, chloropicrin, haloacetic acid, semicarbazide and the like, so that potential threat is caused to human health. Currently, in order to reduce the initial bacteria number on the chicken surface, many manufacturers often use sodium hypochlorite in excess in a precooling pool, which causes a certain hazard. In recent years, with the increasing awareness of consumer health, the use of chemical sterilant has been increasingly limited. Reducing or discarding chemical microbial reduction agents or developing novel healthy and effective chicken carcass microbial reduction technologies has become a new development trend. However, sodium hypochlorite has been shown to have limited effectiveness in disinfecting meat surfaces and interfaces.
The chicken skin has rough surface, complex structure, a large number of hair follicle structures, and the surface layer has hydrophobic fat components, so that the cleaning and disinfection are difficult. In addition, after the pre-cooling link, the hair follicle volume is contracted, and part of the hair follicle is converted into a closed state, so that bacteria are trapped inside the hair follicle, the subsequent entry of a bacteria reducing agent such as sodium hypochlorite is prevented, and the sterilization effect of the bacteria reducing agent is inhibited. The total number of surface colonies of chicken wings treated with 50-100 ppm sodium hypochlorite is reduced by only 0.4log CFU/g. When 200ppm sodium hypochlorite is used for treating the surface of chicken skin, the amount of salmonella enteritidis is reduced by only 0.66log CFU/g, and the chicken quality is seriously affected by the concentration of 200ppm sodium hypochlorite.
Ozone, a non-thermal sterilization technique, has been widely used in the food industry. Researches show that the ozone treatment has obvious bacteria reducing effect, can effectively reduce microorganisms in meat, has high ozone decomposition speed, has no harmful residues after sterilization treatment and does not influence the sense organ. At present, the application of the combined use of ozone water treatment and ultrasonic wave synergistic sodium hypochlorite treatment in the ice fresh chicken sterilization technology has not been reported yet.
Disclosure of Invention
Aiming at the prior art, the invention aims to provide a method for sterilizing chilled chicken.
In order to achieve the above purpose, the invention adopts the following technical scheme:
the invention provides a method for reducing bacteria of chilled chicken, which comprises the following steps:
(1) Soaking the frozen fresh chicken in sodium hypochlorite solution, and performing ultrasonic treatment;
(2) Draining the chicken processed in the step (1), soaking in an ozone water solution, and then draining, packaging in a tray and refrigerating.
Preferably, in the step (1), the concentration of the sodium hypochlorite solution is 45-55mg/L, and the temperature is 0-10 ℃.
The power of the ultrasonic treatment in the step (1) is 5-7kW as a preselection.
Preferably, in step (1), the time of the ultrasonic treatment is 20 to 30 minutes.
Preferably, in the step (2), the concentration of the ozone water is 1.6-4.8mg/L, and the temperature is 4-10 ℃.
Preferably, in the step (2), the soaking time is 5-15min.
Preferably, in step (2), the refrigeration temperature is 2±2℃.
The invention has the beneficial effects that:
the invention combines the ozone water bacteria reduction technology and the ultrasonic synergistic sodium hypochlorite bacteria reduction technology, provides a novel, green and more effective bacteria reduction mode, and effectively prolongs the shelf life of the chilled chicken.
Drawings
Fig. 1: influence of different bacteria reduction modes on sensory quality of drumsticks on day 0 of storage;
fig. 2: influence of different bacteria reduction modes on the sensory quality of the drumstick on the 3 rd day of storage;
fig. 3: influence of different bacteria reduction modes on sensory quality of drumsticks on the 5 th day of storage;
fig. 4: the surface of the chicken samples of different treatment groups belongs to a flora structure on the level and relative abundance thereof; con: a control group; OZ: an ozone water treatment group; SU: ultrasonic synergistic low-concentration sodium hypochlorite treatment groups; SUO: ozone water treatment is combined with ultrasonic synergistic low-concentration sodium hypochlorite treatment group; the numbers represent storage times of 0 days, 3 days, 5 days.
Detailed Description
It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the present application. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
In order to enable those skilled in the art to more clearly understand the technical solutions of the present application, the technical solutions of the present application will be described in detail below with reference to specific embodiments.
The test materials used in the examples of the present invention are all conventional in the art and are commercially available.
Example 1
A method for sterilizing iced fresh chicken comprises the following steps:
(1) Soaking fresh Carnis gallus Domesticus (with skin) in sodium hypochlorite solution with concentration of 45mg/L and temperature of 8deg.C, and ultrasonic treating for 25min; wherein the ultrasonic treatment power is 6KW, and the ultrasonic treatment frequency is 25kHz;
(2) Draining the chicken processed in the step (1), soaking in an ozone water solution with the concentration of 4.8mg/L and the temperature of 8 ℃ for 5min, draining, packaging in a tray, and refrigerating at the temperature of 2+/-2 ℃.
Example 2
A method for sterilizing iced fresh chicken comprises the following steps:
(1) Soaking fresh Carnis gallus Domesticus (with skin) in sodium hypochlorite solution with concentration of 50mg/L and temperature of 8deg.C, and ultrasonic treating for 25min; wherein the ultrasonic treatment power is 6KW, and the ultrasonic treatment frequency is 25kHz;
(2) Draining the chicken processed in the step (1), soaking in an ozone water solution with the concentration of 3.2mg/L and the temperature of 8 ℃ for 10min, draining, packaging in a tray, and refrigerating at the temperature of 2+/-2 ℃.
Example 3
A method for sterilizing iced fresh chicken comprises the following steps:
(1) Soaking fresh Carnis gallus Domesticus (with skin) in sodium hypochlorite solution with concentration of 55mg/L and temperature of 8deg.C, and ultrasonic treating for 25min; wherein the ultrasonic treatment power is 6KW, and the ultrasonic treatment frequency is 25kHz;
(2) Draining the chicken processed in the step (1), soaking in an ozone water solution with the concentration of 1.6mg/L and the temperature of 8 ℃ for 15min, draining, packaging in a tray, and refrigerating at the temperature of 2+/-2 ℃.
Experimental example
1. Experimental method
1.1 grouping of animals
The experiment selects the whole chicken leg of white feather chickens which are slaughtered and segmented and have carcass weight of about 3kg as an experiment raw material after 36 days of a certain food company, wherein the chicken leg is a normal chicken leg without obvious scars and siltation. The whole chicken legs with similar weight are randomly collected by the production line, put into a sterilized packaging bag, transported back to a laboratory within 25min at 0-4 ℃ and immediately subjected to corresponding treatment.
Randomly collecting the left and right chicken whole legs of 36 white feather broilers into four groups: control group (clean water soaking for 5 min), ozone water treatment group (4.8 mg/L ozone water soaking for 5 min), ultrasonic synergistic low-concentration sodium hypochlorite treatment group (soaking in 45mg/L sodium hypochlorite solution and adopting ultrasonic treatment for 25 min), ozone water treatment combined with ultrasonic synergistic low-concentration sodium hypochlorite treatment group (adopting the bacteria reduction method of the embodiment 1). The left and right legs of 9 chickens in each group were respectively subjected to tray packaging after the treatment, and stored under refrigeration (2.+ -. 2 ℃) for 0, 3 and 5 days. And randomly taking out 3 chicken drumsticks from each group at corresponding time points to determine physical and chemical indexes, sensory indexes and microbial indexes, wherein the chicken drumsticks of 1 chicken are the same as the microbial indexes in analysis, the chicken drumsticks of 1 chicken are used for physical and chemical index analysis, and the chicken drumsticks of 1 chicken are used for sensory analysis index determination. This experiment was independently repeated 3 times (n=3). The term "0 day" as used herein means that each index is measured immediately after the treatment.
1.2 measurement items and indices
1.2.1 determination of pH of fresh drumsticks
At various time points during storage, the pH value of the chicken leg meat is measured by using a portable pH meter, and the pH meter is corrected by a two-point method before use (the standard buffer solution with the pH value of 7.00 and the pH value of 4.00 ensures that the correction slope is more than 95%). 6 points are randomly selected on the surface of the whole chicken leg, a pH meter is inserted into the fresh chicken leg, counting is carried out after the reading of the pH meter is stable, and the final result is averaged.
1.2.2 measurement of loss of stored juice from fresh drumsticks
The weight of the treated tray before packaging was designated as W 2 At each storage point, unpacking and taking the weight of the fresh drumstick with the surface juice absorbed by the absorbent paper as W 3 。
Stored juice loss (%) = (W) 2 -W 3 )/W 2 ×100％
1.2.3 determination of volatile basic Nitrogen (TVB-N)
Reference is made to GB5009.228-2016 with a slight modification, and Kjeldahl method is used. 5g of chicken skin and 5g of meat sample without obvious fat adhesion are taken, chopped and put into a 100mL conical flask, 75mL of distilled water is added, and the mixture is uniformly stirred by a glass rod and immersed for 30min, so that the sample is uniformly dispersed in the sample liquid. The impregnated sample was poured into a distillation tube, 1g of magnesium oxide was added, immediately connected to a distiller of a Kjeldahl nitrogen determination instrument, distilled for 180s, and received with 30mL of 20g/L boric acid solution. The mixed solution of methyl red and bromocresol green is used as an indicator, and the titration is carried out by using a hydrochloric acid standard titration solution with the concentration of about 0.01 mol/L. The volume of hydrochloric acid standard titration solution consumed by titration is recorded as V 1 The volume of the hydrochloric acid standard titration solution used for titrating the blank sample is recorded as V 2 。
The content of volatile basic nitrogen (Total volatile base nitrogen, TVB-N) in the sample was calculated as follows:
X＝[(V 1 -V 2 )×C×14]/m×100
wherein:
the content of volatile basic nitrogen in the X-test sample is expressed in milligrams per hundred grams (mg/100 g);
V 1 -the volume of hydrochloric acid standard titration solution consumed for titration of the sample in milliliters (mL);
V 2 -the volume of hydrochloric acid standard titration solution used for titration of blank samples in milliliters (mL);
the concentration of the C-hydrochloric acid standard titration solution is expressed in terms of moles per liter (mol/L);
14-titration of 1.0mL of hydrochloric acid [ c (HCl) =1.000 mol/L ] equivalent nitrogen content of standard titration solution, in grams per mole (g/mol);
m-sample mass in grams (g);
the results of the 100-calculation are converted into conversion coefficients of milligrams per hundred grams (mg/100 g).
1.2.4 determination of lipid Oxidation (TBARS value)
Reference is made to the assay method of Siu et al (1978) and a slight improvement is made. At each storage time point, 2g of chicken skin (removing subcutaneous fat) and 2g of chicken were randomly sheared from the surface of the chicken leg and placed in 100mL test tubes, 20mL distilled water was added to the test tubes, and homogenized for 1min (30 s apart and 30s further) with a homogenizer (T18; IKA, germany). The homogenized homogeneous solution was thoroughly mixed with 20mL of 10% (w/v) trichloroacetic acid solution, and filtered using filter paper, and the filtrate was obtained for use. 4mL of the filtrate was pipetted into the centrifuge tube, 1mL of 60mM thiobarbituric acid solution was added to the centrifuge tube and mixed well, and then the centrifuge tube was placed in a water bath for 90min at 80℃and after 90min, the tube was removed and cooled to room temperature. The cooled solution was transferred to a 96-well plate at 250. Mu.L, and the absorbance of the test solution was measured at 532 nm. The absorbance values obtained were compared with a standard curve of a known concentration of 1, 3-tetraethoxypropane solution, and lipid oxidation values were calculated from the standard curve, and the results were expressed as Mg (MDA)/kg.
1.2.5 sensory evaluation
Reference is made to the method of organoleptic evaluation of iced fresh chicken of Sun Rui (2017) with minor modifications. All samples were coded with random numbers and evaluation scoring was performed immediately after opening the package at each storage time point. Sensory evaluation of the study: the panel consisted of ten individuals experienced in the sensory quality study of chicken. Determining the index and notice of sensory evaluation before the evaluation starts; the members were not evaluated for communication with each other at each evaluation. The samples were subjected to sensory evaluation in a standard evaluation room. The indexes of the sensory evaluation mainly comprise: the chicken skin color of the frozen fresh chicken leg, the whole smell of the frozen fresh chicken leg, the tissue state of the frozen fresh chicken leg and the whole acceptability. The score was 100 points, where a score of 60 was a score limit and a score below 60 was unacceptable, and the higher the score obtained for the indicator, the better the acceptance of the indicator, with specific scoring criteria shown in table 1.
Table 1 sensory evaluation table of fresh frozen drumsticks
1.2.6 microbial count culture
Reference is made to the method of Yang et al (2016) and a slight modification. 10g of chicken skin and meat samples are randomly taken from the frozen fresh chicken leg under the aseptic environment, placed in an aseptic beating bag (BagPageR; interscience, st. Nom, france), 90mL of 0.85% sodium chloride solution (containing 0.01% peptone) is added, and evenly stirred with a beater for 2min at room temperature. Absorbing 1mL of mixed bacterial liquid to make a series of 10-time gradient dilutions, respectively placing the diluted liquid with proper gradient in different culture mediums for plate culture, and making three gradients in parallel. The different selective media and the culture conditions are shown in Table 2.
TABLE 2 Medium and culture conditions for microbial culture
Note that: PCA: lithographic counting agar; MRS: MRS medium.
1.2.7 determination of microbial diversity
(1) Extraction of total bacterial DNA:
taking 10g of chicken skin and meat sample from the surface of fresh chicken leg under the aseptic environment, shearing, fully mixing with 90mL of physiological saline of 0.85% NaCl-0.1% tryptone, and fully beating for 90s in a beater by using a beating bag. Placing 30mL of beating liquid into a centrifuge tube, centrifuging for 5min at 800 Xg and 4 ℃, then placing 15mL of supernatant into another sterilization centrifuge tube, centrifuging for 5min at 12000 Xg and 4 ℃, dissolving the precipitate by using 1.5mL of 0.15mol/L NaCl solution, and centrifuging for 5min at 12000 Xg and 4 ℃, thereby obtaining the precipitate containing the DNA sample. The bacterial DNA extraction kit was used as described, and the total DNA extracted was detected by 0.8% agarose gel electrophoresis and quantified by an ultraviolet spectrophotometer. The method provided by Beijing Nobela source technology Co., ltd. Is adopted, the CTAB or SDS method is adopted to extract the genome DNA of the sample, then agarose gel electrophoresis is utilized to detect the purity and concentration of the DNA, a proper amount of sample DNA is taken out in a centrifuge tube, and the sample is diluted to 1 ng/. Mu.L by using sterile water. Polymerase chain reaction amplification was performed.
(2) PCR amplification
The diluted genomic DNA was used as a template, and specific primers with Barcode were used according to the selection of the sequencing region, new England Biolabs companyHigh-Fidelity PCR Master Mix with GC Buffer and High-efficiency High-fidelity enzyme to perform PCR amplification, thereby ensuring the amplification efficiency and accuracy.
The bacterial 16S rRNA V3-V4 was amplified by polymerase chain reaction using a PCR apparatus with reaction primers 515F (GTGCCAGCMGCCGCGGTAA) and 806R (GGACTACHVGGGTWTCTAAT). The PCR instrument program is set to 98 ℃ for 1min of pre-denaturation, and the thermal cycle is carried out; denaturation at 98℃for 10s, annealing at 55℃for 30s, extension at 72℃for 30s,30 cycles, the last extension being at 72℃for 5min.
(3) Mixing and purification of PCR products
And (3) carrying out equal-concentration sample mixing according to the concentration of the PCR product, fully and uniformly mixing, and then, purifying the PCR product by using agarose gel electrophoresis with the concentration of 1 xTAE of 2%, and tapping to recover a target strip. The product purification kit used was a Thermo Scientific company GeneJET gel recovery kit.
(4) Library construction and on-machine sequencing
UsingThe DNA PCR-Free Sample Preparation Kit library construction kit is used for constructing a library, the constructed library is quantified by Qubit and Q-PCR, and after the library is qualified, novaSeq6000 is used for sequencing on the machine.
2. Experimental results
2.1 Effect of different means of attenuation on the pH of drumstick meat
TABLE 3 influence of different means of attenuation on pH during storage of fresh drumsticks
Note that: A-C indicating the same storage time, the difference between the different treatments reached a significant level (P<0.05)； x-z Indicating that the same treatment regimen, the difference in storage time reached a significant level (P<0.05)。
As can be seen from table 3, during storage, there was no significant difference in pH between the control group and the ultrasonic co-sodium hypochlorite treatment group, and the pH of the ozonated water treatment group and the ozone-combined ultrasonic co-low concentration sodium hypochlorite treatment group were significantly lower than the other two treatment groups, but still in the pH range of normal chicken.
2.2 Effect of different means of attenuation on loss of drumstick storage sap
TABLE 4 influence of different modes of attenuation on storage losses (%) during storage of drumsticks
Note that: A-D indicating the same storage time, the difference between the different treatments reached a significant level (P<0.05)； x-z Representing the same treatment mode, different storagesThe difference in hiding time reached a significant level (P<0.05)。
As can be seen from table 4, the storage loss of the ozone-coupled ultrasound-synergistic low-concentration sodium hypochlorite treatment group was significantly lower than that of the other treatment groups (P < 0.05).
2.3 Effect of different means of attenuation on the volatile basic Nitrogen content in drumsticks
TABLE 5 Effect of different treatments on the volatile basic Nitrogen content (mg/100 g) of fresh drumsticks during storage period
Note that: A-D indicating the same storage time, the difference between the different treatments reached a significant level (P<0.05)； x-z Indicating that the same treatment regimen, the difference in storage time reached a significant level (P<0.05)。
As can be seen from table 5, the volatile basic nitrogen content of the control group and the ultrasonic co-sodium hypochlorite treatment group increased significantly at the later stage of storage, while the TVB-N values of the ozone water treatment group and the ozone-combined ultrasonic co-low concentration sodium hypochlorite treatment group did not increase significantly at the later stage of storage, indicating that the growth and metabolic activities of microorganisms could be effectively inhibited after the ozone treatment. On the 5 th day of storage, the TVB-N value of the ozone-combined ultrasonic-assisted low-concentration sodium hypochlorite treatment group is only 8.45mg/100g, which is obviously lower than that of the other three treatment groups, and the result shows that the ozone-combined ultrasonic-assisted sodium hypochlorite treatment on the frozen fresh drumsticks can play a good role in delaying spoilage, so that the frozen fresh drumsticks can keep good freshness.
2.4 Effect of different modes of attenuation on the extent of lipid oxidation in drumsticks
TABLE 6 influence of different treatments on the acid number of thiobarbituric acid in fresh drumsticks (mg MDA/kg) during storage
Note that: A-D indicating the same storage time, the difference between the different treatments reached a significant level (P<0.05)； x-z Indicating that the same treatment regimen, the difference in storage time reached a significant level (P<0.05)。
As can be seen from table 6, the TBARS value of the ozone-combined ultrasonic-synergistic low-concentration sodium hypochlorite treatment group was significantly higher than that of the other three treatment groups, because ultrasonic-synergistic sodium hypochlorite treatment had promoted lipid oxidation of the frozen fresh drumsticks, and the cavitation effect of the ultrasonic was damaging the muscle fiber tissues of the drumsticks, further aggravating lipid oxidation of the frozen fresh drumsticks after being immersed in ozone water. However, the increase in lipid values of the ozone-coupled ultrasonic-assisted low-concentration sodium hypochlorite treatment group was not shown in the sensory-rated odor scores, indicating that the ozone-coupled ultrasonic-assisted sodium hypochlorite treatment, while increasing lipid oxidation of chicken skin and chicken, did not adversely affect the sensory and quality of the frozen chicken leg.
2.5 Effect of different means of attenuation on sensory evaluation of drumsticks
As can be seen from fig. 1-3, the sensory scores of the frozen fresh drumsticks of each treatment group continued to decrease with prolonged storage time. At the end of storage, the microbial numbers of the three bacteria-reducing treatment groups are obviously lower than those of the control group, and the growth and propagation of microorganisms lead to the generation of volatile substances such as amine substances, so that the odor freshness of the frozen drumsticks is reduced, and the odor of the frozen drumsticks of the control group is unacceptable. This indicates that the control group was not acceptable to the consumer already at day 5 of storage. The odor scores of the three attenuated treatment groups were significantly higher on both days 3 and 5 of storage than the control group.
The appearance freshness, the tissue state, the overall acceptability and the odor freshness scores of the frozen drumsticks are the same, and the appearance freshness, the tissue state and the overall acceptability of the three bacteria-reducing mode treatment groups are obviously higher than those of the control group until the end of storage (5 days). Overall, the combination of ozone and ultrasonic in combination with sodium hypochlorite treatment provides a better maintenance of the organoleptic qualities of the chilled drumsticks.
2.6 influence of different bacteria reduction modes on microbial indicators on the surface of the fresh drumsticks
2.6.1 Effect of different bacteria reduction modes on the total number of bacterial colonies on the surface of a fresh drumstick
TABLE 7 Effect of different treatments on the surface colony count (log CFU/g) of fresh drumsticks during storage period
Note that: A-D indicating the same storage time, the difference between the different treatments reached a significant level (P<0.05)； x-z Indicating that the same treatment regimen, the difference in storage time reached a significant level (P<0.05)。
As can be seen from Table 7, the interaction between the treatment regimen and the storage time has a significant effect on the colony count of the fresh drumsticks (P<0.05). With the increase of the storage time, the total number of colonies per sterilization mode treatment group increased significantly. On day 0, the three bacteria reduction modes all obviously reduce the initial colony count on the surface of the frozen fresh drumstick, and compared with a control group, the ultrasonic synergistic sodium hypochlorite treatment group reduces the CFU/g by 0.97log (the initial microorganism number is 10 from the control group) 5.72 CFU/g reduction to treatment group 10 4.75 CFU/g, the bacterial reduction amount reaches 89.3 percent, and the ozone water treatment group is reduced by 1.26log CFU/g compared with the control group (the initial microorganism number is 10 from the control group) 5.72 CFU/g reduction to treatment group 10 4.46 CFU/g, the bacterial reduction amount reaches 94.5 percent; ozone-coupled ultrasound in combination with low concentration sodium hypochlorite treatment reduced 1.47log CFU/g compared to control (initial microbial count from 10 for control) 5.72 CFU/g reduction to treatment group 10 4.25 CFU/g, the bacteria reduction amount reaches 96.6 percent). On day 3 of storage, the total number of colonies of the control group was 6.46log CFU/g, and the level of spoilage had been reached, but the total number of colonies of the three attenuated treatment groups were all below 6log CFU/g, and the fresh drumsticks were still fresh. On day 5 of storage, the total number of colonies of the ultrasonic synergistic sodium hypochlorite treatment group had also exceeded 6log CFU/g, whereas the total number of colonies of the ozone water treatment group and the ozone-combined ultrasonic synergistic low-concentration sodium hypochlorite treatment group were 5.96log CFU/g, respectively (the number of microorganisms was 10 from the control group 7.19 CFU/g reduction to treatment group 10 5.96 CFU/g, 93.4% reduced) and 5.79log CFU/g (microbial count from 10 for control group) 7.19 CFU/g drop10 low to treatment group 5.79 CFU/g, reduced by 95.5%), no spoilage, and effectively prolonged shelf life of the frozen fresh drumsticks for 2 days in both bacteria reduction modes. But the total number of bacterial colonies of the frozen fresh drumsticks in the ozone-ultrasonic combined synergistic sodium hypochlorite treatment group is lower than that in the ozone water treatment group, and the fresh-keeping effect is better.
2.6.2 Effect of different means of attenuation on the number of E.coli on the surface of fresh drumsticks
TABLE 8 influence of different treatments on the number of E.coli on the surface of a fresh drumstick (log CFU/g) during storage
Note that: A-D indicating the same storage time, the difference between the different treatments reached a significant level (P<0.05)； x-z Indicating that the same treatment regimen, the difference in storage time reached a significant level (P<0.05)。
As can be seen from table 8, the interaction between the treatment regimen and the storage time has a significant effect on the growth of enterobacteria (P < 0.05). The number of the enterobacteria on the surface of the frozen fresh drumsticks of each treatment group shows a trend of obviously rising along with the extension of the storage time, and the number of the enterobacteria between the ozone water treatment group and the ozone-combined ultrasonic synergistic low-concentration sodium hypochlorite treatment group is not obviously different, but the number of the enterobacteria of the frozen fresh drumsticks of the two bacteria reduction modes is obviously lower than that of the other two treatment groups. During storage, the number of enterobacteria in the ozone water treatment group and the ozone-combined ultrasonic synergistic low-concentration sodium hypochlorite treatment group is not more than 4log CFU/g all the time. While the control group had 4.55log CFU/g on day 3 and exceeded 5log CFU/g by day 5, significantly higher than the other treatment groups. Ozone and ultrasonic are combined to cooperate with the low-concentration sodium hypochlorite treatment group to obviously reduce the number of enterobacteria on the surface of the frozen fresh drumstick, and the method shows better bacteria reduction advantage.
2.6.3 influence of different bacteria reduction modes on the quantity of lactic acid bacteria on the surface of a fresh drumstick
TABLE 9 Effect of different treatments on the surface lactic acid bacteria count (log CFU/g) of fresh drumsticks during storage
Note that: A-D indicating the same storage time, the difference between the different treatments reached a significant level (P<0.05)； x-z Indicating that the same treatment regimen, the difference in storage time reached a significant level (P<0.05)。
From Table 9, it can be seen that the lactic acid bacteria count of each treatment group showed an increasing trend as the storage time of the ice fresh drumsticks was prolonged. Compared with a control group, the three bacteria reduction modes obviously reduce the number of lactic acid bacteria on the surface of the frozen fresh drumsticks. On day 0, the ozone water treatment group and the ultrasonic synergistic sodium hypochlorite treatment group respectively reduce the lactobacillus count of the frozen fresh drumsticks from 4.87log CFU/g to 4.00log CFU/g and 4.09log CFU/g; the ozone and ultrasonic combined low-concentration sodium hypochlorite treatment group has the best bacteria reduction effect, and the lactobacillus count of the frozen fresh drumsticks is reduced by 0.99log CFU/g. The number of the enterobacteria in each treatment group is obviously increased from the 3 rd day to the 5 th day, the number of the lactic acid bacteria in the ozone-combined ultrasonic synergistic low-concentration sodium hypochlorite treatment group is only 4.36log CFU/g after the 5 th day of storage, and the bacteria reduction efficacy is optimal.
2.7 influence of different bacteria reduction modes on microbial diversity of the surface of the fresh drumsticks
As can be seen from fig. 4, each of the bacteria-reduced treatment groups effectively reduced the bacterial diversity as compared with the control group, wherein the ozone water treatment combined with the ultrasonic synergistic sodium hypochlorite treatment (SUO) had the best reducing effect. This demonstrates that ozone water treatment combined with ultrasound in combination with sodium hypochlorite treatment has a great advantage in terms of the bacteria reduction effect. Meanwhile, shewanella abundance in each group of samples gradually increased with prolonged storage time. However, ozone water treatment combined with ultrasound synergistic sodium hypochlorite treatment significantly reduced the abundance of shiwanella compared to the control, ozone water treatment (OZ) and ultrasound synergistic low concentration sodium hypochlorite treatment (SU) treatment groups. Shewanella is the main spoilage bacteria in poultry, and reducing the abundance of this bacteria helps to prolong chicken shelf life.
The foregoing description is only of the preferred embodiments of the present application and is not intended to limit the same, but rather, various modifications and variations may be made by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.
Claims (7)
1. The method for sterilizing the iced fresh chicken is characterized by comprising the following steps of:
(1) Soaking the frozen fresh chicken in sodium hypochlorite solution, and performing ultrasonic treatment;
(2) And (3) soaking the chicken processed in the step (1) in an ozone water solution, and then draining, packaging in a tray and refrigerating.
2. The method for reducing bacteria in a fresh chicken according to claim 1, wherein in the step (1), the concentration of the sodium hypochlorite solution is 45-55mg/L and the temperature is 0-10 ℃.
3. The method for reducing bacteria in iced fresh chicken according to claim 1, wherein the power of the ultrasonic treatment in the step (1) is 5-7kW.
4. The method for reducing bacteria in iced fresh chicken according to claim 1, wherein in the step (1), the time of the ultrasonic treatment is 20-30min.
5. The method for reducing bacteria in a fresh chicken according to claim 1, wherein in the step (2), the concentration of the ozone water is 1.6-4.8mg/L and the temperature is 4-10 ℃.
6. The method for reducing bacteria in iced fresh chicken according to claim 1, wherein the soaking time in the step (2) is 5-15min.
7. The method for sterilizing chilled chicken according to claim 1, wherein in the step (2), the refrigerating temperature is 2.+ -. 2 ℃.
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