Revealthe the composition and properties of bacteria and yeast in Weining ham.

Abstract: The study of the microorganisms in Weining ham, Guizhou, was studied for separation and identification of salt, nitrite and low temperature resistanceThe results showed that the co-isolated strains included strains of staphylococcus (), strain sucrose (), strain lactic acid tablet succobacteria (), strain smoothed false silk yeast () and strain near-smooth false silk yeast ();
Weining ham is a famous traditional fermented meat products in Guizhou has a history of many yearsWeining ham from the highland lean meat type good seed "Ujin pig" hind legs processing the pig leg muscle tight muscle fiber quality for the processing of Weining ham provides superior raw materialsAt present, Weining ham is still extended to the family workshop-style model in the annual winter solstice after the production of natural fermentation productionThe microorganisms in natural fermentation mainly come from the complex types of microorganisms in raw meat and the surrounding ham and are susceptible to harmful microorganisms, so the product quality is seriously affected by the environmental impact and even causes the failure of the fermentation process to cause the ham to corrupt and spoil
It has become a trend to use artificial inoculation to control the production process in modern processing of fermented meat productsCompared with natural fermentation, artificial inoculation fermentation can not only maintain product quality stability, shorten fermentation time, but also ensure product safety and uniform production standards to extend product shelf lifeTherefore, it is very important to isolate and identify the beneficial microorganisms in Weining hamIn this study, weinn ham as the research object using dash culture method to separate the microorganisms and identify the salt, nitrite and low temperature resistance characteristics of the study at the same time in order to reveal the advantages of the microbial distribution in Weining ham to achieve its industrial production to provide a scientific theoretical basis;
Materials and methods Materials and reagents Materials and reagents The weining ham of the mature year was purchased from the farmers' market of the Hui Hmong Autonomous County of the Weining Yi ethnic group and placed in a cold-temperature transport box with ice packs to the laboratory
Sodium nitrite Shandong Yantai Jiajing Food Industry Co., Ltd.; Sodium chloride National Pharmaceutical Group Chemical Reagents (Shanghai) Co., Ltd.; Grameen DyeD Beijing Soleibao () Technology Co., Ltd.; Bacterial Genome Extraction Kit Beijing Tiangen Biochemical Technology Co., Ltd.; Column Yeast Genome Extraction Kit Bioengineering (Shanghai) Co., Ltd
Medium glycol sodium chloride agar ( ) liquid medium (beef dip paste, 䏡 protein, glycol, sodium chloride, phenol red, distilled watersterilization) homemade;
Add agar to each of the above liquid mediums to obtain the corresponding solid mediumSolid media, solid media, malt solid medium scored aureus, lactic acid bacteria, yeast separation culture
Instruments and Equipment Biosecurity Cabinet Singapore Yisi Hi-Tech Co., Ltd.; Micro-desktop high-speed centrifuge Hunan Xiangyi Laboratory Instrument Development Co., Ltd.; Visible Spectrophotometer Shanghai Precision Science Instrument Co., Ltd.; Series of several-point water bath pot Jiangsu Jintan City Jincheng Guosheng Experimental Instrument Factory; portable stainless steel pressure steam steriliser Shanghai Sanshen Medical Device Co., Ltd.; ; Electrophoremeter Beijing 61 Biotech Co., Ltd.; sterile homogenizer Ningbo Xinzhi Biotech Co., Ltd.; polymerase chain reaction () gene amplification instrument USA; fluorescent microscope Japan Company
MethodS Sample treatment In sterile conditions, the sample was cut from the surface and interior of the Vening ham with a sterile knife and placed in a sterile bag containing sterile physiological saline Place the sterile bag in a sterile homogenizer and beat it to a homogenous fluid
The separation and purification of the strains take the above-mentioned homogenous liquid added to the aseptic physiological saline moderately diluted and absorbed the coating in the solid medium, solid medium and malt juice solid medium in the culture of the Using the vaccination ring to pick a single colony using flat dash method for dashing culture repeat operation until the acquisition of pure-bred culture The purified strains were inoculated on a beveled medium and stored at the degree C
Species Identification Morphological Observation The characteristic analysis of colonies morphological features: observing and recording the surface morphology, color, transparency and other characteristics of the colonies
Analysis of individual morphological characteristics: selecting bacterial bacteria for Gramey dyeing post-mirror examination, yeast through the water-immersion method to observe and record the form of bacterial cell
Serial identification The bacterial and yeast genomes are extracted by reference to the bacterial and yeast genome s/he s/hess and is kept back-up under -C conditions Bacterial genes are amplified with universal primers: upstream primers ('') downstream primers ('') The internal transcription interval ( ) of yeast is amplification of the general primer as: upstream primer ('') downstream primer (''' ) The reaction system is: s, template, primer, aseptic, mixed Reaction conditions: the pre-variable s.C
The increase in yield was tested by agarose gel electrophoresis and sent to Bioengineering (Shanghai) Co., Ltd for sequencing The database is used to analyze the homologousness of genes to obtain different kinds of sequences with higher homologousity of the gene sequence of the destination Using software to build a system development tree at the same time consider the characteristics of strain colonies and bacterial morphology to confirm the name of the strain
Microbial characteristics study Salt-resistant properties Absorbing activated progenitors of mycobacterium inoculated in selective liquid media with a mass fraction, and selective liquid medium, the culture of the medium was studied with unvaccinated media as a blank control Bacteria and yeast are cultured to determine the density of light at the wavelength of the culture liquid ( )
Nitrite-resistant properties Absorbtheactivated generation of mycobacteria liquid inoculation in the addition of the amount of respectively, the selective liquid medium in the culture of the former as a control group Bacteria and yeast are cultured and measured in culture The tolerance factor is defined as the ratio of the experimental group to the control group
Low temperature resistance characteristics Absorbtheactivated generation of myobacteria liquid inoculated in selective liquid medium in the culture of the culture group under the condition s.C Bacteria and yeast are cultured and measured in culture The tolerance factor is the ratio of the experimental group to the control group
Data processing Use software for data profiling using software to map The resultofs of each experiment repetition are expressed as average s/standard deviation
Results and Analysis The separation and identification of microorganisms in Weining ham were isolated and purified by the dominant microorganisms in Weining ham, and the co-numbers of the strains were numbered separately, and the types of microorganisms in Weining ham were identified by morphological analysis and sequence analysis respectively
Morphological analysis from the table can be seen: the isolated purification of the strain of bacteria are Gram-positive () colony form round edge neatly paired, piles or arranged into grapes; Pictured is the image of the bacterial Gramch dye mirror examination pictured is the yeast mirror image
Sequence identification Extraction strain genomes are obtained and amplification is carried out The molecular mass of the sample-amplified increase is about the sample, the molecular mass of the amplified increase is about, and the amplified strip is clear and bright The molecular mass of all strips is in line with theoretical expectations
The sequence of isolated strains was analyzed by the database to determine the results of the sespecies identification A separate strain of Weining ham can be seen from the table include strains of strain staphylococcus (), strain xylitose staphylococcus (), strain lactic acid tablet succobacteria (), strain smoothed false silk yeast () and strain near smooth fake silk yeast ()
Comparing the three domestic ham Xuanwei ham is the most significant feature is the presence of a large number of salt-resistant bacteria on the surface of its dominant flora including staphylococcus and microccicocci Huang Aixiang found that the dominant bacteria in Yunnan dried pickled ham are mainly ma gastric staphylococcus, epidermal staphylococcus, Staphylococcus aureus, staphylococcus, and mutant micrococcal bacteria among them are the dominant bacteria in the process of cloud leg processing The relatively small number of yeasts detected in the samples were identified as red yeast, Debali yeast, Hanson yeast, false silk yeast, bent false silk yeast and hidden ball yeast Jiang Yunsheng and other research found that in the natural fermentation process, such as ham, bacteria, bacteria, yeast is the dominant bacteria group Yu Xiang found that the advantages of bacteria in Jinhua ham produced by modern fermentation technology were lactic acid bacteria followed by staphylococcus, and the yeasts that dominated the fungus were mainly Onobi fake silk yeast, white buller spore yeast and so on Foreign well-known pickled ham in the ham fermentation and maturation process of the advantagebacteria including staphylococcus genus and micrococcal genus from the ham surface samples can be isolated from the milk coccorgen bacteria from the internal samples can be isolated salt-resistant bacteria The yeast strain obtained from the separation from the ham surface has been identified as belonging to the Debali yeast genus, the false silk yeast genus and the Hansen yeast genus
Most of the microorganisms identified in this study are more common in domestic and foreign hams (Xuanwei ham, such as ham, Jinhua ham, ham, ham, ham, ham) such as staphylococcus, xylitose staphylococcus and lactic acid tablets However, there are several microorganisms such as smoothed false silk yeast and near-smooth false silk yeast reported less may be due to differences in microbial types in different hams in different production environments Nearly smooth false silk yeast was established as a new classification unit at the seed level Liang Hui and others isolated from the traditional wax fish to obtain the scented yeast of the plant, which is near smooth fake silk yeast Guo Zhuang and others isolated from raw film beef sauce to smooth fake silk yeast and found that their role in making beef sauce production film in ham needs to be further studied
Study on the properties of isolated microorganisms in Weining ham From the isolated species of bacteria, each selected strain represents the strain (、、、、) to conduct salt, nitrite and low temperature resistance
The study of salt resistance is an important factor in the selection of fermented meat products fermentation agent, which has an important influence on the sensory quality and biosecurity of fermented meat products The strains screened require normal growth at at least the salt content Therefore, the effect of salt content on the growth of strains was studied
It can be seen from the graph that the growth of each strain showed a downward trend with the increase of the content of the medium The maximum growth of each strain at the time of content indicates that high content inhibits the growth of the strain Among them, the growth of lactic acid tablet cocci in the content decreased significantly () more than When the content increased, xylitose staphylococcus basically stopped growing staphylococcus, smoothed false silk yeast and near-smooth fake silk yeast growth decreased but still can grow salt resistance is better
Pan Ming from such as ham separation, purification to get ear staphylococcus, epidermis staphylococcus aureus species staphylococcus aureus show that the appropriate concentration can promote the growth of staphylococcus staphylococcus in which the most salt-resistant when the most salt-resistant content of its inhibitory xyloccus stapy staphylococcus is slightly worse but under the content can still grow its salt resistance than stapy staphylococcus
The study of nitrite-resistant properties is an important additive in the production of meat products The meat industry usually requires strains to grow well under sodium nitrite Therefore, the effect of adding sodium nitrite on the growth of strains was studied
It is possible to grow strains of bacteria in mediums with sodium nitrite content Among them, the tolerance coefficient of staphylococcus aureus is close to indicating that its growth is almost immune to nitrite additions significantly higher than other strains( xylitose staphylococcus, lactic acid tablet succosiella and near-smooth pseudo-silk yeast growth is inhibited in the amount of sodium nitrite added but the tolerance coefficient can reach the above nitrite resistance second; and the lowest tolerance coefficient of smoothy yeast (to nitrite) is the least resistant to nitrite
Li Xinwei and others in Jianmen ham staphylococcus bacteria, lactic acid bacteria in nitrite addition sphinches when the growth results show that the strains have a certain nitrite resistance characteristics of which Staphylococcus (and) to nitrite has a strong tolerance and lactic acid bacteria (and) tolerance is relatively weak PanMing from such as ham separation, purification of the results of ear staphylococcus, epidermal staphylococcus aureus, xylitose staphylococcus aureus and de Ballyhanson yeast have better nitrite resistance characteristics of which Staphylococcus aureus nitrite resistance is the best Consistent with the results of this study
Low temperature resistance research temperature plays an important role in the production of fermented meat products In general, the pickled temperature of fermented meat products is the most suitable fermentation temperature of the temperature of the product s.C In addition, Chinese ham production cycle is longer its marinating process is generally not less than Therefore, this study explores the effectofs of low temperature (C) on the growth of strains It is shown that the strains can maintain a certain amount of growth when the culture temperature is reduced to c of c, but there are significant differences in the degree of influence of different strains from low temperature The tolerance coefficient of the strains of bacteria is, and the tolerance of the bacteria to the low temperature is in turn, the tolerance coefficient of staphylococcus aureus in the masculine staphylococcus aureus is significantly higher than that of other bacteria The highest tolerance coefficients of smoothed false silk yeast and near-smooth false silk yeast at the temperature of the culture of c of c were respectively and showed that they could still grow well under low temperature conditions
Liu Xiaoqiang isolated from Yunnan ham to obtain the mutant micrococcal and sucrose staphylococcus bacteria are tolerable and nitrite The strains of glycoccus and Bacillus tin can also be tolerated and nitrites are also tolerated and nitrites are not resistant to low temperatures The xylifi staphylococcus aureus was consistent with Liu Xiaoqiang's findings, which were consistent with nitric acid resistance, while lactic acid tablets were less salinizing than those
Conclusion The microbes isolated from Weining ham were determined to be strains of staphylococcus, xylitose staphylococcus, lactic acid tablets, smoothed false silk yeast and near-smooth false silk yeast after colony characteristics, individual morphological analysis and sequence identification Among them, staphylococcus, smoothed fake silk yeast and near-smooth fake yeast are salinizing better lactic acid tablet cocci salinity resistance is slightly worse and xylitose staphylococcus bacteria can hardly grow under the content Staphylococcus aureus is the worst resistant to the best level of sodium nitrite tolerance for the best smooth false silk yeast The resistance to low temperature sesame and near-smooth fake silk yeast at the culture temperature of the degree c c was significantly higher than that of strains of bacteria with the highest tolerance coefficient of staphylococcus Therefore, staphylococcus and near-smooth fake silk yeast salt resistance, nitrite resistance and low temperature resistance can be better adapted to different external environment
This study reveals that the composition and characteristics of bacteria and yeast in Weining ham have expanded the beneficial bacteria reservoir of meat products to provide a theoretical basis for the localization research and development of subsequent meat product fermentation agents The influence of microorganisms on the flavor of ham and other qualities in the fermentation process of Weining ham has yet to be further studied
References: Slightly citation format: Zhang poetic Tang Nan Huangpan and other Wei Ning ham in the separation identification of microorganisms and its tolerable characteristics of meat research Source: Meat Research.