发酵肉中凝固酶阴性葡萄球菌快速鉴定

介绍一种凝固酶阴性葡萄球菌(CNS)快速鉴定方法:“两步一补法”。并用此方法鉴定传统发酵肉中CNS的分布情况。CNS菌株先经二步鉴定:第一步4h,包括:蕈糖、尿素酶、碱性磷酸酶;第二步24h,包括:鸟氨酸脱羧酶、新生霉素敏感性、磷霉素敏感性、厌氧生长。极少数不能鉴定的CNS菌株,用补充试验完成结果。本方法能鉴定出发酵肉中CNS的22个种及2个亚种,两步法能鉴定出发酵牛肉中77%以上常见CNS菌种,仅23%的菌株需用补充实验证实。     

发酵香肠风味物质气质分析及与发酵时间的关系

分析取样量、萃取时间、萃取温度3个因素对发酵香肠中挥发性成分萃取效果的影响,并对发酵6,9h的香肠中的香气成分进行了比较分析。结果表明:发酵香肠中风味物质的抽提条件为样品质量3.0g,萃取时间50min,萃取温度60℃。从发酵6,9h的香肠中分别鉴定出54,20种香气成分;酯类物质是发酵6h肠中主要挥发性成分,而发酵9h肠中主要成分为醇类,其中70.9%是乙醇,乙醇谱峰远高于其他化合物的谱峰,掩盖了其他化合物的谱峰。发酵6h与发酵9h的香肠中香气成分组成及相对含量具有明显的差异性。就风味物质而言,发酵6h的效果优于发酵9h的。     

发酵香肠益生型乳酸菌发酵剂的筛选

从主要自然发酵肉制品中分离、纯化得到77株耐酸耐胆汁的乳酸菌,并进一步对分离菌株进行了耐高酸(pH2.5)、0.3%胆盐的生长实验.25株在pH值2.5,培养3 h后表现出较高存活性,21株经低酸预处理后在0.3%胆盐存在的情况下生长良好.最后运用API 50CHL对筛选菌株进行了鉴定.这些菌株可作为进一步筛选发酵香肠优良乳酸菌种的出发菌株,以便筛选品质优良的益生性乳酸菌,作为新型发酵香肠发酵剂.     

不同发酵香肠中微球菌与葡萄球菌的筛选与鉴定

144株分离色拉米发酵香肠的微球菌科菌株被鉴定.所用的发酵香肠来自三个地区:新疆、山东、湖南,每个地区选择两个厂,香肠出自制作的的三个不同阶段.其中木糖葡萄球菌是主要菌种(95%),经过生化反应鉴定出共有12种典型的木糖葡萄球菌,2号和5号两种菌占大多数(38.8%,33.6%).根据观察,木糖葡萄球菌5号,显示硝酸盐还原酶和脲酶活性,适度的蛋白质和脂肪分解能力,产生乙偶姻,适合做发酵剂.     

发酵香肠乳酸菌生理生化特性的研究

采用中国传统发酵香肠作为样品,从中分离纯化出乳酸菌,且对其做生理生化特性鉴定,以对发酵香肠的菌株进一步的筛选,从中选出不产黏液、发酵葡萄糖不产气、不产色素、不产H2S、不具有氨基酸脱羧酶、不产氨、产酸能力强的菌株作为发酵香肠中的优良的乳酸菌,最后测定优势菌株24h的生长曲线和pH值变化曲线.     

产低温几丁质酶菌株的筛选、鉴定与产酶条件优化

目的:筛选产低温几丁质酶菌株,并对其进行鉴定及发酵条件优化。方法:本实验以渤海海域海泥为样品,以胶体几丁质为唯一碳源,通过平板筛选法筛选产低温几丁质酶细菌,对其进行形态学鉴定和分子生物学鉴定,并对其发酵产酶条件进行了单因素优化。结果:菌株鉴定结果为发光杆菌(Photobacterium sp.),单因素优化结果为:胶体几丁质12.0 g/L、酵母膏6.0 g/L、发酵温度15 ℃、初始pH7.0、转速为220 r/min、装液量75 mL/250 mL、接种量1%、发酵时间96 h,酶活力达4.566 U/mL,比优化前提高389.91%。结论:为下一步酶学性质研究及低温几丁质酶在工业中的应用提供参考。     

发酵香肠葡萄球菌、微球菌生理生化特性的研究

采用中国传统发酵香肠作为样品,从中分离纯化出葡萄球菌,且对其做生理生化特性鉴定,以对发酵香肠的菌株进一步的筛选,从中选出不产粘液、发酵葡萄糖不产气、不产色素、不产H2S、不具有氨基酸脱羧酶、不产氨、15℃能还原硝酸盐、pH值4.5务件下生长的菌株作为发酵香肠中的优良葡萄菌株.     

发酵香肠中益生性乳杆菌的分离及筛选

利用PH2.5和1%胆盐的MRS液体培养基,对自然发酵香肠中耐低酸耐胆盐的乳杆菌进行富集,从中分离纯化得到184株乳杆菌。通过一级筛选:24h产酸、耐盐耐硝性、产粘液、产过氧化氢等实验,筛选出20株符合发酵香肠乳酸菌发酵剂基本要求的菌株。通过二级筛选:耐低酸能力、耐胆汁酸盐能力和抑菌能力,最终筛选出了性能优良乳杆菌3株,可作为益生型发酵剂用于发酵香肠生产。文中还对所获得菌株的相关发酵性能进行了测定。     

酸菜中降解亚硝酸盐乳酸菌的筛选、鉴定及其在发酵香肠中的应用

目的:依据多项评价指标筛选亚硝酸盐降解能力强且适用于肉制品发酵的乳酸菌。方法:首先采用添加0.3%Ca CO3的MRS培养基从15份不同产地的农家自制酸菜中筛选乳酸菌,其后以降解亚硝酸盐能力和肉制品发酵剂所需要求为筛选指标对初筛所得乳酸菌进行复筛,并对其进行菌种的分子鉴定。最后以筛选到的菌株为发酵剂制作单菌株发酵香肠,通过检测香肠在发酵和贮藏期间的Na NO2含量来进一步验证所得菌株的实际降解亚硝酸盐能力。结果:筛选到1株降解亚硝酸盐能力优良(降解率为78.77%)且适于肉制品发酵的乳酸菌菌株,16S r DNA全序列分析鉴定为鼠李糖乳杆菌(Lactobacillus rhamnosus GG,ATCC 53103)。香肠发酵实验结果显示,在发酵结束时香肠的Na NO2含量为12.2 mg/kg,低于国家标准。同时,微生物指标检测结果证明了此条件下生产的发酵香肠的质量是安全可靠的。结论:鼠李糖乳杆菌能有效降低发酵香肠中的Na NO2含量,可用于开发成用于生产低亚硝酸盐的健康、安全的肉品发酵剂。      

中式发酵干香肠中乳酸菌的分离筛选、鉴定和培养

从烟台喜旺中式发酵干香肠中分离筛选出适合当地特色的、具有自然发酵香肠风味的微生物菌株,采用系统分类鉴定方法对筛选出的乳酸菌进行初步鉴定,鉴定出5种乳酸杆菌:L1为德氏乳杆菌德氏亚种,L2为德氏乳杆菌保加利亚亚种,L3为米酒乳杆菌,L4为食品乳杆菌,L5为戊糖乳杆菌。此外,采用均匀设计确定了这5种乳酸杆菌的增殖培养基的组成和最佳配比,并通过混合培养得出混合培养的最佳配比。     

Selection of Lactobacillus strains from fermented sausages for their potential use as probiotics

A rapid screening method was used to isolate potentially probiotic Lactobacillus strains from fermented sausages after enrichment in MRS broth at pH 2.5 followed by bile salt stressing (1% bile salts w/v). One hundred and fifty acid- and bile-resistant strains were selected, avoiding preliminary and time-consuming isolation steps. Strains were further characterized for survival at pH 2.5 for 3 h in phosphate-buffered saline and for growth in the presence of 0.3% bile salts with and without pre-exposure at low pH. Twenty-eight strains showed a survival >80% at pH 2.5 for 3 h; moreover, most of the strains were able to grow in the presence of 0.3% bile salts. Low pH and bile resistance was shown to be dependent on both the species, identified by phenotypic and molecular methods, and the strain tested. This is the first report on the direct selection of potentially probiotic lactobacilli from dry fermented sausages. Technologically interesting strains may be used in the future as probiotic starter cultures for novel fermented sausage manufacture.     

清酒乳杆菌对发酵风干肠品质的影响

该研究采用清酒乳杆菌（Lactobacillus sakei，L. sakei）作为发酵菌株制备发酵风干肠，研究其对发酵风干肠品质的影响，以自然发酵产品作为对照组，在发酵成熟过程中的第0、3、6、9天测定产品的菌落总数、乳酸菌数、pH值、水分含量、水分分布、嫩度、色差、过氧化物值（Peroxide Value，POV）、硫代巴比妥酸值（Thiobarbituric Acid Value，TBARS）、脂肪酸含量和感官评分等指标，研究发酵菌株对风干肠品质的影响。试验表明，L. sakei接种组的乳酸菌数和菌落总数显著高于对照组（P<0.05），L*值、a*和b*显著低于对照组（P<0.05）；L. sakei接种组在发酵终点时，水分含量降至32.15%，pH值降至4.41，保证了发酵风干肠的安全性；POV值和TBARS值显著低于对照组（P<0.05），表明L. sakei具有较好的抑制脂肪氧化作用；L. sakei接种组的总体可接受性更高，发酵后期酯类物质种类和含量均高于对照组。综上，L. sakei的添加改善了产品的品质和风味。     

肉用发酵菌株的筛选及其在低盐发酵香肠中的应用

该研究以蛋白酶、脂肪酶活性和抑菌活性筛选肉用发酵菌株,通过分子生物学技术对其进行鉴定,并将其应用于低盐发酵香肠中,通过感官、理化及微生物指标分析评价其应用效果。结果表明,筛选鉴定得到两株适用于肉类发酵、安全且具有蛋白酶及抑菌活性的菌株,分别为植物乳杆菌（Lactiplantibacillus plantarum）BR-12和木糖葡萄球菌（Staphylococcus xylose）H-1,其中菌株BR-12还具有脂肪酶活性,两菌株间无拮抗作用。菌株BR-12与H-1按2∶1和1∶5的比例接种制备的低盐发酵香肠pH下降迅速,肠杆菌数与挥发性盐基氮含量较低,感官评分较高,硬度适中,弹性好,呈特有的玫瑰红色,且两菌株接种比例为2:1时,品质更好,说明菌株BR-12和H-1复配发酵有利于提高产品安全性,改善低盐发酵香肠滋气味,可用于低盐香肠发酵。     

Colour parameters of Turkish-type fermented sausage during fermentation and ripening

Colour parameters of Turkish-type fermented sausage during fermentation and ripening were evaluated. CIE x y, L^* a^* b^*, C^* h°, pigment nitrosation (NI) and pigment discoloration (RSI) were measured. Sausage processing had four stages consisting of first and second fermentation steps and then two consecutive ripening steps. Variations of colour parameters with processing indicated that Turkish-type fermented sausage processing might be divided into two well defined phases from the colour point of view. Optimum colour was obtained at the end of the first ripening step.     

接种发酵对发酵牛肉香肠品质及多肽抗氧化活性的影响

以接种复配发酵剂作为实验组,以自然发酵作为对照组,通过测定pH值、色差、质构、亚硝酸盐含量等理化指标探究接种发酵对牛肉发酵香肠品质的影响;通过提取香肠多肽并测定多肽的1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基、2,2'-联氮-双-(3-乙基苯并噻唑啉-6...     

高抗氧化活性肉用乳杆菌的筛选鉴定及其在发酵香肠中的应用

目的 筛选高抗氧化活性肉用乳杆菌,并应用到发酵香肠中。方法 以肉品发酵剂标准、自由基清除率、耐过氧化氢能力和抗氧化酶活性为指标进行菌株筛选,经形态学、生理生化及16S rDNA对菌株进行鉴定,通过硫代巴比妥酸值、羰基和巯基值探究菌株对发酵香肠的抗氧化作用。结果 菌株CC-3符合硝酸盐还原酶阳性、氨基酸脱羧酶阴性等肉用发酵剂标准；该菌株具有较高的自由基清除能力,其发酵上清液DPPH.和OH.清除率分别为98.43%、42.75%,菌体细胞.清除率为75.56%；具有较高的抗氧化酶活性,其细胞破碎液T-SOD活力和CAT活力分别为45.17±0.29、58.50±0.28 U. mgprot_^-1,发酵上清液GSH-Px活力为98.16±0.65 U.mgprot_^-1。菌株CC-3鉴定为植物乳杆菌。接种菌株CC-3的发酵香肠与其他组比较在发酵结束时TBARS值和羰基值最低,分别为0.22±0.01、1.57±0.06 nmol.mg_^-1；巯基值最高,为0.05±0.01 nmol.g_^-1。结论 菌株CC-3符合肉用发酵剂标准且具有高抗氧化活性,可以抑制发酵香肠脂肪氧化和蛋白氧化,具有重要的开发潜力。     

A novel polymerase chain reaction (PCR) - denaturing gradient gel electrophoresis (DGGE) for the identification of Micrococcaceae strains involved in meat fermentations. Its application to naturally fermented Italian sausages

A new molecular method consisting of polymerase chain reaction (PCR) amplification and denaturing gradient gel electrophoresis (DGGE) of a small fragment from the 16S rRNA gene identified the Micrococcaceae strains isolated from natural fermented Italian sausages. Lactic acid bacteria, total aerobic mesophilic flora, Enterobacteriaceae and faecal enterococci were also monitored. Micrococcaceaea control strains from international collections were used to optimise the method and 90 strains, isolated from fermented sausages, were identified by biochemical tests and PCR-DGGE. No differences were observed between the methods used. The results reported in this paper prove that Staphylococcus xylosus is the main bacterium involved in fermented sausage production, representing, from the tenth day of ripening, the only Micrococcaceaea species isolated.     

Antilisterial activity of lactic acid bacteria isolated from "Alheiras" (traditional Portuguese fermented sausages): In situ assays

A total of 226 lactic acid bacteria (LAB) isolated from “Alheira”, a traditional Portuguese fermented sausage, were screened for antagonistic activity against some pathogenic microorganisms, including Listeria monocytogenes. The objective was to isolate LAB with antibacterial activity from “Alheiras” and to select strains that could be used in “Alheira” production. Isolates displaying antibacterial activity against Listeria innocua and L. monocytogenes were investigated for the nature of the antibacterial compounds active against these microorganisms. Results showed that two LAB cultures retained activity in the supernatants after neutralization and catalase treatment. These two strains were both identified as Pediococcus pentosaceus. The final aim of this work was to test the antilisterial activity of these two strains during storage of “Alheira mass” (sterilized), at 4 °C. The growth of L. innocua population was significantly suppressed in the paste of “Alheira” when the samples were co-inoculated with the LAB strains, in comparison with the paste only inoculated with L. innocua or co-inoculated with a bacteriocin negative strain of Ped. pentosaceus (ca. 1 × 10⁷ CFU/g after 28 days of incubation).     

Strain dependent expression of stress response and virulence genes of Listeria monocytogenes in meat juices as determined by microarray

A subgenomic array, encompassing 54 probes targeting genes responsible for virulence, adhesion and stress response in Listeria monocytogenes, was used in order to study their expression in food systems. RNA extracted from L. monocytogenes inoculated in BHI and in situ (i.e. in minced meat and fermented sausage juices) and incubated at 4 °C, was hybridized on the array and the results obtained were compared in order to understand the effect that the food juice has on the expression. Three different strains of L. monocytogenes were tested, in order to determine the effect of the strain provenience. As determined by cluster analysis, each strain behaved in a different way when inoculated in food juices. The goal was to respond to acidic and osmotic stresses encountered in the food, particularly in the fermented sausage juice. No differences in the expression profile between the three strains were observed, when they were inoculated in BHI. On the other hand, in the meat and sausage juices, the iap, gadC and gadE genes, together with different internalin encoding genes, were significantly differentially expressed in the three strains.