基于刃天青的电化学方波伏安法快速鉴别大肠杆菌的耐药性

目的：评价5株食源性大肠杆菌临床分离株对环丙沙星、庆大霉素和氨苄西林的抗生素耐药性。方法：基于细菌能量代谢过程中的电子传递理论,引入刃天青作为氧化还原探针,建立了一种基于刃天青介导的大肠杆菌耐药性快速鉴别的电化学方波伏安法（square wave voltammetry,SWV）。结果：在1.69×10²～1.69×10⁸ CFU/mL范围内,大肠杆菌浓度与刃天青氧化峰电流（I）之间存在较好的线性关系,回归方程为Y=-0.156 0X-2.965 5,R²=0.977 5;在进行大肠杆菌耐药性测定时,参考美国临床和实验室标准协会制定的药敏试验标准,以刃天青的氧化峰电流（I₀）和大肠杆菌直接与刃天青作用后得到的氧化峰电流（I_-a）作为对照,测定大肠杆菌用抗生素处理后与刃天青作用得到氧化峰电流（I_+a）,引入细菌活性值V_ETA作为电化学方波伏安法鉴别细菌耐药性的指标,建立了刃天青介导的大肠杆菌电化学快速药敏试验方法;运用建立的方法检测了5种食源性大肠杆菌临床分离株对3种抗生素的耐药性,以V_ETA=50作为判断细菌敏感和耐药的临界值,均得到与纸片扩散法结果相符的药敏试验结果,而纸片扩散法报告结果需18 h,该法仅需2 h。结论：建立的基于刃天青介导的大肠杆菌耐药性快速鉴别的电化学方波伏安法能够准确且快速检测5株食源性大肠杆菌临床分离株对环丙沙星、庆大霉素和氨苄西林的抗生素耐药性。     

生奶利色唑林(刃天青)试验和细菌检验对照的研究

利色唑林(分子式C_(12)H_7NO_4),又名刃天青,也称树脂天青,是一种氧化还原剂,具有相当高的著色价,付与新鲜牛奶一种特殊的蓝色。某些微生物在奶中繁殖,能分泌一种具有还原作用的酶——还原酶,可     

婴幼儿配方奶粉原料乳形式的技术经济分析

在阐述婴幼儿配方奶粉原料乳形式的基础上,从新鲜程度、营养成分和成本费用三个方面比较生鲜乳与复原乳的技术经济性,分析了大量使用复原乳对我国奶粉进口和奶牛养殖业造成的影响。结果表明,采用生鲜乳为原料在新鲜程度和营养成分上显著优于复原乳,并有利于我国奶牛养殖业发展。最后从宣传生鲜乳优势、建立生鲜乳生产婴幼儿配方奶粉制度和降低奶牛养殖成本三个方面提出推动生鲜乳全面取代复原乳成为婴幼儿配方奶粉原料乳的政策建议。     

不同贮存条件对原料乳质量的影响

研究了原料乳质量和贮存温度及贮存时间之间的关系.通过菌落总数的检测和酸度的测定,探讨了不同贮存温度随时间对原料乳中菌落总数和酸度的影响,旨在为优质原料乳的贮运提供参考.结果表明,在4℃贮存条件下,原料乳存放36 h,对质量没有较大的影响;在原料乳温度达到10～15℃的时候,存放时间不宜超过12 h.     

不同体细胞数的原料乳品质变化的比较研究

通过分析比较5组不同体细胞数(SCC)原料乳的主要成分和特性(蛋白质、脂肪含量、酪蛋白的含量等)、脂肪分解情况(脂肪酶活力、游离脂肪酸及其占总脂肪的比例)以及蛋白质水解程度(水溶性氮、非蛋白氮占总氮的比例以及酪蛋白构成)的变化情况,探讨不同SCC对原料乳品质的影响。结果表明：当SCC小于4.0×105个/mL时,原料乳的成分没有显著的差异(P＞0.05);随着SCC的增大,原料乳的脂肪水解程度增强,但SCC小于4.0×105个/mL的两组原料乳的脂肪水解程度没有显著差异(P＞0.05);不同SCC原料乳的酪蛋白构成表现不同,其蛋白质水解程度(WSN/TN,NPN/TN)随SCC的增大而增加,但SCC在4.0×105 个/mL以下的两组原料乳的蛋白水解程度没有显著差异(P＞0.05)。     

原料乳中嗜冷菌菌群多样性及分析方法研究进展

嗜冷菌是原料乳中主要危害性腐败菌群之一,其种类繁多,菌群结构和多样性受到多种因素影响,这为原料乳中优势嗜冷菌的鉴定带来了困难.结合国内外文献,综述了储藏温度、储藏时间、地域和季节等因素对原料乳中嗜冷菌菌群结构和多样性的影响,原料乳中嗜冷菌可能的外界来源.评述了高通量测序、随机扩增多态性DNA、变性凝胶电泳等现代分子生物...     

一种发酵乳中乳酸菌总数快速检测技术的研究

以还原法为理论基础,开发并建立一种应用于发酵乳饮料成品乳酸菌数指标的快速检测方法。本方法应用仪器记录了样品加入反应试剂后的颜色变化(RGB),通过数理统计分析,采用斜率法,以R值斜率表示样品颜色的变化速度。本研究对发酵乳样品进行了预处理,以符合刃天青还原法的反应条件。以GB 4789.35《食品安全国家标准食品微生物学检验乳酸菌检验》为标准,建立了R值斜率与乳酸菌菌落总数之间的标准方程,经验证本方法检测结果与GB 4789.35《食品安全国家标准食品微生物学检验乳酸菌检验》检测结果无显著差异。与现有标准方法相比,在时效性、运营成本等方面具有较大优势。     

加拿大原料乳安全生产及其质量控制体系

食品安全成了当今世界极度关注的重点.随着我国乳业的迅速发展,我国乳品生产安全的问题也越来越引起国家职能部门、专家和企业的关注.而乳制品的质量在很大程度上取决于原料乳的质量.加拿大等国在原料乳质量控制领域中进行了深入而实用的研究工作,制定了一套以HACCP为基础的有效的质量安全操作规范,极大的促进了原料乳的发展和提高了原料乳的质量.加拿大牛奶生产过程中的质量控制主要包括员工培训,危害性及其来源,牛奶生产设施,水和饲料,动物健康和生物防护,医药、治疗和后遗症预防,挤奶管理,利用乳牛生产牛肉,纠偏措施和肉乳生产纠偏指南等10个部分.本文重点介绍原料乳生产过程中的危害及其控制.原料乳生产过程中的危害性污染源主要来源于化学污染、生物污染和物理污染.其质量控制主要通过最佳管理实践(Best Management Practices,BMPs)、危害分析和关键控制途径(Hazard Analysis Critical Control Point(HACCP)Approach)、在HACCP之内的关键控制点(Critical Control Points(CCPs)within HACCP)和标准的操作程序(Standard Operating Procedures,SOPs)实施.     

双蛋白发酵乳制品的研制开发及其营养价值

双蛋白发酵乳是以大豆分离蛋白、鲜牛奶为主要原料,经保加利亚乳杆茵和嗜热链球菌发酵而成的一种高蛋白营养保健饮品,具有延年益寿防衰老的保健作用.详细介绍了双蛋白发酵乳的生产工艺,并讨论了影响其稳定性的因素.     

预测微生物学在低温储存原料乳中的应用

随着冷藏设备的广泛应用,嗜冷菌及其耐热代谢产物成为影响原料乳质量和乳制品质量的主要因素。以奶站的新鲜原料乳为研究对象,研究春季原料乳中总菌落和嗜冷菌在不同储存温度下的生长情况。使用Gompertz模型建立4℃~14℃储存的原料乳中总菌落和嗜冷菌的生长动力模型,模型可以有效的模拟原料乳中微生物的生长情况。低温储存的原料乳中总菌数与嗜冷菌有一定的相关性。     

基于16S rDNA高通量测序方法比较新疆西北部地区乳品中微生物的多样性

为更加全面地了解新疆特色乳品中微生物多样性,比较不同动物来源的原奶和酸奶的细菌群落结构,运用高通量测序技术,对乳品中细菌16S r DNA V4-V5区测序,进而对新疆克州和塔城地区牛奶、驼奶、马奶、羊奶、酸牛奶、酸驼奶和酸马奶7种乳品中细菌群落组成和多样性进行分析。研究共获得539 557条有效序列,379个OTU。多样性分析表明,原奶样品中细菌Shannon-Wiener指数明显高于酸奶样品。微生物群落组成分析发现,不同乳品之间菌群组成差异较大。7种乳品中的菌群均以厚壁菌门和变形菌门为主,但原奶样品主要以变形菌门为主,而酸奶样品主要以厚壁菌门为主。在属水平上,牛奶主要以假单胞菌属(Pseudomonas)为主,驼奶主要以埃希菌属-志贺菌属(Escherichia-Shigella),马奶主要以明串珠菌属(Leuconostoc)为主,羊奶中的优势菌属为乳球菌属(Lactococcus),而酸牛奶、酸驼奶和酸马奶都是以乳杆菌属(Lactobacillus)为优势菌属。不同动物来源的原奶和酸奶样品中的微生物多样性存在显著差异,并且原奶中检测到的环境污染菌和致病菌(或条件致病菌)的丰度也相对较高。本研究结果将为准确评估乳品中的微生物群落对新疆地区少数民族健康的影响提供一定的数据基础。     

Antibacterial effect of the lactoperoxidase system on psychotrophic bacteria in milk.

Activation of the antibacterial lactoperoxidase system in milk, i.e. increasing the thiocyanate concentration to 0.25 mM and adding an equimolar amount of H2O2, results in a substantial reduction of the bacterial flora and prevents the multiplication of psychrotrophic bacteria for up to 5 d. This treatment has no effect on the physico-chemical properties of milk and does not lead to the accumulation of resistant bacteria. The practical application of the lactoperoxidase system in prolonging the storage period of raw milk at low temperatures is discussed.     

Evaluation of biogenic amines and microbial counts throughout the ripening of goat cheeses from pasteurized and raw milk

The effect of the hygienic quality of milk on changes in microbial counts and biogenic amine content was evaluated during ripening of goat cheeses manufactured from pasteurized and raw milks at 1, 14, 30, 60 and 90 d. The original milk, rennet, curd and whey were also included in the study. The pH, salt content and extent of proteolysis in the cheese were also evaluated. Spermidine and spermine were the main amines in raw milk, while they were minor amines in cheeses. Other amines increased markedly during ripening, tyramine being the main amine in cheese made from raw milk and cadaverine and putrescine in those produced from pasteurized milk. Enterobacteriaceae counts decreased during ripening whereas those of lactic acid bacteria increased, especially lactobacilli and enterococci. Cheese made from raw milk showed higher microbial counts during ripening than those made from pasteurized milk, especially for Enterobacteriaceae and enterococci, counts being 2 or 3 log units higher. Raw milk cheese showed remarkably higher biogenic amines compared with pasteurized milk cheeses. Therefore, pasteurization of milk causes a decrease in final biogenic amine content of cheese as a result of the reduction of its microbial counts.     

Effect of high pressure processing on the safety,shelf life and quality of raw milk

High pressure processing (HPP) was investigated as an alternative to standard raw milk processing. Different pressure levels (400–600 MPa) and exposure times (1–5 min) were tested against artificially inoculated pathogenic E. coli, Salmonella and L. monocytogenes. HPP effectively inactivated bacterial concentration by 5 log CFU/ml. The most effective HPP conditions in terms of pathogen reduction were subsequently utilised to determine the effect of pressure on microbiological shelf life, particle size and colour of milk during refrigerated storage. Results were compared to pasteurised and raw milk. HPP (600 MPa for 3 min) also significantly reduced the total viable counts, Enterobacteriaceae, lactic acid bacteria and Pseudomonas spp. in milk thus prolonging the microbiological shelf life of milk by 1 week compared to pasteurised milk. Particle size distribution curves of raw, pasteurised and HPP milk, showed that raw and HPP milk had more similar casein and fat particle sizes compared to pasteurised milk. The results of this study show the possibility of using HPP to eliminate pathogens present in milk while maintaining key quality characteristics similar to those of raw milk.     

The effects of carbon dioxide addition to cheese milk on the microbiological properties of Turkish White brined cheese

This study invest0igated the effect of CO₂ added to achieve three pH levels: pH 6.1, pH 6.2 and pH 6.3 for treatments X, Y, Z, respectively, on some microbiological properties of Turkish White (TW) brined cheese. For each pH, four batches of cheese were produced from: raw milk with no added carbon dioxide (UR), raw milk with carbon dioxide (TR), pasteurised milk with no carbon dioxide addition (UP) and pasteurised milk with carbon dioxide addition (TP). The microbiological analysis of TW brined cheeses was carried out for 90 days of maturation period. Total aerobic mesophilic bacteria, mesophilic lactic acid bacteria, yeasts and moulds and coliform group were determined in control and CO₂ treatment groups. Mesophilic bacteria count was determined as 5.14, 5.29, 5.67 log cfu/g for pH 6.1, 6.2 and 6.3, respectively, in CO₂‐treated raw milk cheeses. Yeasts and moulds reduction increased significantly by applying CO₂ (P < 0.01). For TW cheese samples, the most significant microbial inactivation was detected at sample groups of pH 6.1.     

Inactivation of microorganisms naturally present in raw bovine milk by high‐pressure carbon dioxide

This study assessed the inactivation of microorganisms naturally present in raw bovine milk by high‐pressure carbon dioxide (HPCD) at 10–30 MPa and 20–50 °C for 20–70 min. The log reduction of microorganisms increased as raw bovine milk was exposed to higher pressures and temperatures and longer treatment times. The maximum reduction of aerobic bacteria (AB) was 4.96‐log at 25 MPa and 50 °C for 70 min. At lower temperatures and treatment times, a complete inactivation of yeasts and moulds (Y&M) and coliform bacteria (CB) was obtained at 25 MPa. Changes in microorganisms naturally present in raw bovine milk during storage were also assessed. There were 1.83‐log survival of AB, 0.65‐log survival of Y&M and a complete inactivation of CB in raw bovine milk when subjected to HPCD at 25 MPa and 40 °C for 50 min. Moreover, the AB, Y&M and the CB in raw bovine milk exhibited insignificant alterations during storage at 4 °C for 15 days, indicating a potential capability of HPCD to extend the shelf life of milk.     

Effect of CO2 addition to raw milk on proteolysis and lipolysis at 4 degrees C

Fresh raw milks, with low (3.1 x 10(4) cell/ml) and high (1.1 x 10(6) cells/ml) somatic cell count (SCC), were standardized to 3.25% fat, and from each a preserved (with 0.02% potassium dichromate) and an unpreserved portion were prepared. Subsamples of each portion were carbonated to contain 0 (control, pH 6.9) and 1500 (pH 6.2) ppm added CO2, and HCl acidified to pH 6.2 Milk pH was measured at 4 degrees C. For the preserved low- and high-SCC milks, two additional carbonation levels, 500 (pH 6.5) and 1000 (pH 6.3) ppm, were prepared. Milks were stored at 4 degrees C and analyzed on d 0, 7, 14, and 21 for microbial count, proteolysis, and lipolysis. The addition of 1500 ppm CO2, but not HCl, effectively delayed microbial growth at 4 degrees C. In general, in both the low- and high-SCC unpreserved milks, there was more proteolysis and lipolysis in control and HCl acidified milks than in milk with 1500 ppm added CO2. Higher levels of proteolysis and lipolysis in the unpreserved milks without added CO2 were related to higher bacteria counts in those milks. In preserved low- and high-SCC milks, microbial growth was inhibited, and proteolysis and lipolysis were caused by endogenous milk enzymes (e.g., plasmin and lipoprotein lipase). Compared with control, both milk with 1500 ppm added CO2 and milk with HCl acidification had less proteolysis. The effect of carbonation or acidification with HCl on proteolysis in preserved milks was more pronounced in the high SCC milk, probably due to its high endogenous protease activity. Plasmin is an alkaline protease and the reduction in milk pH by added CO2 or HCl explained the reduction in proteolysis. No effect of carbonation or acidification of milk on lipolysis was observed in the preserved low- and high-SCC milks. The CO2 addition to raw milk decreased proteolysis via at least two mechanisms: the reduction of microbial proteases due to a reduced microbial growth and the possible reduction of endogenous protease activity due to a lower milk pH. The effect of CO2 on lipolysis was mostly due to a reduced microbial growth. High-quality raw milk (i.e., low initial bacteria count and low SCC) with 1500 ppm added CO2 can be stored at 4 degrees C for 14 d with minimal proteolysis and lipolysis and with standard plate count < 3 x 10(5) cfu/ml.     

单分子实时测序技术研究不同来源原料乳中群落组成及多样性

目的 采用三代单分子实时测序法(single molecule real time sequencing,SMRT)研究北京和内蒙古两地原料乳中的细菌群落组成及多样性,解析其携带污染菌的差异,为原料乳质量及风险评估提供基础数据.方法 收集北京和内蒙古两地春夏季和秋冬季的原料乳样品20份,采用SMRT技术对其进行基于16...     

乳及乳制品中嗜冷菌的多样性及其腐败危害研究进展

乳是一种营养丰富的物质,同时也是微生物生长繁殖的理想培养基。生乳的质量是影响乳制品产业链的关键因素,随着低温储存和冷链运输技术的发展,生乳中大部分细菌的生长受到抑制,但嗜冷菌的生长并未受到抑制,并逐步成为生乳中的优势菌。生乳在冷藏运输或储存期间,嗜冷菌依旧可以生长繁殖,其分泌的蛋白酶和脂肪酶可耐高温,经过巴氏杀菌或超高温灭菌处理后依旧保持活性,因此,了解嗜冷菌的多样性及其产生的酶对提高乳及乳制品质量、减少腐败和浪费具有重要作用。本文通过介绍乳及乳制品中嗜冷菌污染现状及腐败危害研究,旨在为乳及乳制品行业的风险评估提供背景信息,从源头控制嗜冷菌对生乳的浪费,保证乳及乳制品的品质。     

High-pressure processing of Turkish white cheese for microbial inactivation

High-pressure processing (HPP) of Turkish white cheese and reduction of Listeria monocytogenes, total Enterobacteriaceae, total aerobic mesophilic bacteria, total molds and yeasts, total Lactococcus spp., and total Lactobacillus spp. were investigated. Cheese samples were produced from raw milk and pasteurized milk and were inoculated with L. monocytogenes after brining. Both inoculated (ca. 10(7) to 10(8) CFU/g) and noninoculated samples were subjected to HPP in a high-pressure food processor at 50 to 600 MPa for 5 and 10 min at 25 degrees C. Reductions in L. monocytogenes, total aerobic mesophilic bacteria, Lactococcus spp., and Lactobacillus spp. in both pasteurized- and raw-milk cheese samples and reductions in total molds and yeasts and total Enterobacteriaceae counts in raw-milk cheese samples increased with increased pressure (P < or = 0.05). The maximum reduction of the L. monocytogenes count, ca. 4.9 log CFU/g, was obtained at 600 MPa. Because of the highly inhibitory effect of pasteurization, the total molds and yeasts and total Enterobacteriaceae counts for the cheese samples produced from pasteurized milk were below the detection limit both before and after HPP. There was no significant difference in inactivation of L. monocytogenes, total aerobic mesophilic bacteria, Lactococcus spp., and Lactobacillus spp. under the same treatment conditions for the raw milk and pasteurized milk cheeses and for 5- and 10-min treatment times (P > 0.05). No significant change was detected in pH or water activity of the samples before and after HPP. Our findings suggest that HPP can be used effectively to reduce the microbial load in Turkish white cheese.