不同介质中甲型副伤寒沙门氏菌的热失活特性

研究甲型副伤寒沙门氏菌在不同介质中热失活规律,对接种10~8 CFU/g甲型副伤寒沙门氏菌的不同介质进行55、63、72℃热处理,测定处理后样品中甲型副伤寒沙门氏菌的菌体浓度。应用Dose Resp模型拟合在3种温度下志贺氏菌的动力学模型,用D值(decimal reduction time)表示甲型副伤寒沙门氏菌在不同介质中的耐热情况。结果表明:甲型副伤寒沙门氏菌的热失活曲线运用Dose Resp模型拟合较好,相关系数均在0.97以上。甲型副伤寒沙门氏菌在不同介质中的D值不同,在蛋白质和脂肪含量较高的灌肠肥瘦肉1∶1(m/m)中D值较大,相对耐热。55℃条件下热处理25.0 min,63℃热处理2.03 min,72℃热处理0.31 min后,一般肉制品中甲型副伤寒沙门氏菌均能被杀死。     

超高压协同温度处理对绿色魏斯氏菌的失活动力学

为了建立超高压协同温度处理对绿色魏斯氏菌在低温烟熏火腿中的失活模型,采用40、45、50 ℃结合 200～500 MPa对接种108～109 CFU/mL绿色魏斯氏菌的烟熏火腿进行5、10、15、20、25、30 min的超高压处理,同 时做相同条件的离体实验进行对比。采用薄层平板计数,以时间为横坐标,亡菌数量级为纵坐标作失活曲线,选择 一级线性模型和非线性模型（Weibull和Logistic）进行模型的拟合,用模型的评价因子相关系数R2、准确因子Af和 均方差验证方程的拟合度。结果表明：经热压结合处理后,接种于烟熏火腿中的绿色魏斯氏菌比相同条件下离体状 态的绿色魏斯氏菌的失活更缓慢,可能因为火腿基质中存在一些大分子物质（如蛋白质、脂肪等）对菌体的保护作 用。Logistic方程最适合描述绿色魏斯氏菌在两种状态下的失活情况,为预测绿色魏斯氏菌在烟熏火腿中的失活提 供了理论依据。     

含肉桂醛猪肉糜中金黄色葡萄球菌热失活模型的建立

研究了添加不同质量分数的肉桂醛(0,0.1%,0.5%和1.0%)后猪肉糜中金黄色葡萄球菌的热失活规律。在肉糜中添加不同浓度的肉桂醛后进行热处理(60～75℃),用选择性培养基进行活细胞计数。结果表明,不同温度条件下,肉桂醛对金黄色葡萄球菌的热失活有明显的促进作用,随着温度和肉桂醛浓度的升高,金黄色葡萄球菌的耐热性逐渐降低。在75℃条件下,肉桂醛使肉糜中金黄色葡萄球菌完全热失活所需的时间从5 min减少到3 min。将试验结果进行动力学模型拟合,多项式拟合作为初级模型,能准确地描述添加肉桂醛后肉糜中金黄色葡萄球菌的热失活规律。在二级模型的拟合中,线性模型具有更高的拟合系数值。通过扫描电镜观察肉桂醛处理条件下金黄色葡萄球菌细胞的形态变化,表明肉桂醛可破坏金黄色葡萄球菌的细胞膜,从而导致胞质泄露而死亡。     

小肠结肠炎耶尔森菌在猪肉中失活/存活模型的初步研究

采用预测微生物学的基本方法和程序,研究小肠结肠炎耶尔森菌在特定条件下的生长规律,并用CurveExpertl.38软件作为辅助工具,从中选取了Linear模型对实验数据进行拟合.初步建立了肉汤中小肠结肠炎耶尔森菌55、60℃的热失活模型,以及猪肉中小肠结肠炎耶尔森菌的速冻失活模型和冷冻存活模型.     

牛肉中沙门氏菌热失活模型的建立

为了建立牛肉中沙门氏菌的热失活动力学模型,将4种不同血清型的沙门氏菌混合菌液接种到牛肉表面,将接种后的牛肉分别在55、57.5、60、62.5和65 ℃下进行加热处理。不同温度下的沙门氏菌热失活曲线用Weibull模型进行了拟合,判定系数(R2)分别为0.993(55 ℃)、0.984(57.5 ℃)、0.999(60 ℃)、0.999(62.5 ℃)和0.998(65 ℃)。进一步建立了温度对Weibull一级失活模型参数(b)影响的二级模型,即ln(b)=0.47T-28.07。用58.5和64 ℃下实际的沙门氏菌存活数对所建的模型进行验证,准确度(A_f)和偏差度(B_f)分别为1.071和1.056,0.998和1.002,均在可接受范围内。本研究所建立的模型能较好的模拟不同温度(55~65 ℃)对牛肉中沙门氏菌热失活的影响。     

反转录-环介导等温扩增技术快速检测志贺氏菌

建立了反转录-环介导等温扩增技术的志贺氏菌快速检测方法。针对志贺氏菌特异保守基因ipa H设计多套引物,建立优化了的反应体系,并评价其准确性、特异性、灵敏度。以人工污染脱脂乳样品比较RT-LAMP与RT-PCR的灵敏度。结果表明,在65℃等温条件下,RT-LAMP反应可在30 min内完成。在活菌/损伤菌模型中,该方法表现出比国标法更好的准确性。在23株细菌标准菌株中仅对4株志贺氏菌有特异性检出。志贺氏菌RNA模板的检测灵敏度为7 fg/μL。人工污染脱脂乳样品检测灵敏度达到40 CFU/g,比RT-PCR方法高出2个数量级。表明所建立的志贺氏菌RT-LAMP扩增方法可以准确的检测志贺氏菌,同时具有快速、特异、灵敏的优势,可用于志贺氏菌的快速筛查和现场监控。     

冷熏三文鱼单增李斯特菌热灭菌规律及其品质变化动力学研究

为了给冷熏三文鱼热灭菌提供理论依据,试验研究了热处理对冷熏三文鱼中单增李斯特菌失活及品质变化影响。将接种了一株单增李斯特菌(ATCC19113)菌液的鱼肉密封于毛细管中,分别在58℃、60℃、62℃、64℃和66℃条件下加热处理18 min、8.8 min、4 min、2 min和1 min,考察不同温度条件下存活菌数变化。采用铝制密封加热单元(直径35 mm×高6 mm)研究了冷熏三文鱼(直径30 mm×高6 mm)在50℃、57℃、64℃和70℃条件下加热处理120 min、90 min、60 min和40 min时的品质变化。结果表明:单增李斯特菌的热失活曲线遵循对数线性关系(R2≥0.94);冷熏三文鱼品质指标加热失重率、a*值、b*值及剪切力遵循1级反应动力学规律,L*值遵循0级反应动力学规律;冷熏三文鱼色值a*与单增李斯特菌热失活密切相关,色值a*品质劣变曲线与单增李斯特菌(ATCC19113)死亡曲线重叠区域为热处理温度与时间组合可操作范围,此范围可同时满足杀菌及品质保持要求。     

热死环丝菌生长预测模型的建立

针对引起冷却肉腐败的优势微生物热死环丝菌,以分离得到的S-8菌株作为受试菌株,研究了0～10℃低温条件下热死环丝菌S-8的生长情况。利用SAS程序拟和不同温度条件下的生长情况,经过比较发现,Gompertz模型比线性模型能更好地拟合热死环丝菌的生长,从而得到了其生长的Gompertz模型参数;利用平方根模型对其的最大比生长速率平方根-温度(U(1/2)-T)进行拟合,得到热死环丝菌S-8生长的二级模型:U(1/2)=0.1192(T+6.00),T∈[0,10];利用培养基数据,冷却肉产品数据和温度波动条件下的数据对所得到的二级模型进行验证,计算得到总的偏差因子和准确因子分别为0.982和1.223,结果表明二级模型能真实快速有效地预测冷链条件下冷却肉中热死环丝菌的生长。     

温度和盐含量对三文鱼中单增李斯特菌热失活的影响

主要考察菌株、温度、盐含量对三文鱼中单增李斯特菌热失活动力学的影响。菌株编号为CICC 21632、CICC 21633、CICC 21635、CICC 21639的4株单增李斯特菌分别接种至盐含量为0, 1.5%, 3.0%和4.5%的三文鱼中,并于不同温度(57.5℃～67.5℃)条件下测定D值。结果表明,单增李斯特菌的热抗性(log D)受菌株、温度、盐含量,以及因素间交互作用的影响显著;菌株CICC 21632为4株受试菌株中热抗性最高, Z值为6.02℃,其热抗性随着盐含量增加而线性增加。动力学分析结果和建立的模型可用于水产品加工行业设计合适的热处理规程,以消除三文鱼中的单增李斯特菌,并确保其微生物性安全。     

耐热乳酸菌热致死动力学模型研究

测定了从云南传统乳制品乳扇中筛选的2种6株耐热乳酸菌在60℃和65℃热处理条件下的菌株存活情况.发现E.faeciumgrx28的耐热能力最强,D60为6.45min,D65为1.32 min与其他菌株有显著差异.分别基于Linear,Weibull和Log-logistic动力学模型,对各耐热乳酸菌在不同热处理条件下的活菌数量的变化情况使用lstopt1.0软件进行数据的曲线拟合及相关系数的分析,确定在60℃的热处理条件下,Weibull模型更适合描述菌株的残活量,而在65℃的热处理条件下,Logistic模型与菌株的存活曲线更为接近,得到各菌株在不同处理条件下的动力学方程.结果表明,乳酸菌在不同热处理条件下,乳酸菌的死亡变化并不相同.     

超高压协同温度处理对烟熏火腿中单增李斯特菌生长预测模型的建立

为了建立超高压协同温度处理烟熏火腿中单增李斯特菌生长预测模型。本研究分别以20、30、40、50℃结合600MPa对接种106cfu/g单增李斯特菌烟熏切片火腿进行5min的超高压处理,4℃贮藏条件下,每10d测定处理后样品中单增李斯特菌的数量。结果表明:当超高压协同处理的温度高于40℃时显著延长了单增李斯特菌在烟熏火腿修复生长的延滞期,所得生长曲线用修正的Gompertz模型进行了拟合,在35、45d进行验证,建立了烟熏火腿中单增李斯特菌在超高压协同温度处理后的生长预测模型,经验证其精确因子(Af)、偏差因子(Bf)、根平均方差(RMSE)和决定系数(R2)均在较好范围内,能较好的模拟单增李斯特菌的生长情况,为低温肉制品中单增李斯特菌的控制提供理论参考。     

THERMAL RESISTANCE, SURVIVAL AND INACTIVATION OF ENTEROBACTER SAKAZAKII (CRONOBACTER SPP.) IN POWDERED AND RECONSTITUTED INFANT FORMULA

Enterobacter sakazakii has recently been recognized as an opportunistic foodborne pathogen, and dry infant formula serves as the mode of transmission. The objectives of this study were to investigate the heat resistance, survival and inactivation under room and refrigeration temperatures storage of dry and reconstituted infant formula milk (IFM). E. sakazakii strains (eight strains) showed a wide variability in heat resistance at different temperatures (55, 60 and 63C). The D-values at 55C ranged from 1.51 to 14.83 min, at 60C from 0.17 to 2.71 min and at 63C from 0.05 to 0.88 min. The calculated z values for the studied E. sakazakii strains ranged from 3.76–10.11C. Microwave oven heating of 60-mL portions of reconstituted IFM for 40–50 s was effective in eradicating inoculated E. sakazakii. Storing powdered IFM for 15 days at 4C resulted in at least a 1-log reduction in E. sakazakii strains, whereas storing reconstituted IFM at 4C for 2 weeks resulted in more than a 2-log reduction in E. sakazakii.

PRACTICAL APPLICATIONS

This study shows that E. sakazakii strains differ widely in their heat resistance. No differences were observed between biofilm formers and nonformers in terms of heat-resistance in thermal inactivation kinetics experiments. Conventional high temperature short-time pasteurization processes are considered sufficient to inactivate all E. sakazakii strains, and a household microwave oven (40–50 s for 60-mL portions) can be used to inactivate E. sakazakii if present in reconstituted infant formula milk (IFM). Growth of E. sakazakii can be inhibited in powdered and reconstituted IFM by refrigeration. Also, it is recommended that reconstituted IFM be discarded or refrigerated if not immediately consumed. The probiotic L. acidophilus ATCC 4356 was not effective in inhibiting E. sakazakii in powdered or reconstituted IFM.     

应用超高压手段延长低温烟熏火腿的货架期

为揭示超高压处理后低温烟熏火腿货架期、品质及营养指标的变化,以400MPa和600MPa的压力在22℃条件下对切片包装后的烟熏火腿进行10min超高压处理,以未经高压处理样品作对照,4℃贮藏条件下,于0、1、30、90d测定超高压处理样品中的各腐败微生物总数,同时对超高压处理烟熏火腿的脂肪氧化程度、游离脂肪酸含量以及部分感官指标进行评定。结果表明：超高压能够有效杀灭烟熏火腿中的肠杆菌、热杀索丝菌、假单胞菌以及霉菌和酵母,乳酸菌中的部分属较为耐压,是低温烟熏火腿贮藏后期的优势腐败菌;超高压处理组比未处理组样品的饱和脂肪酸含量略有上升,不饱和脂肪酸含量略有下降,且随压力升高饱和脂肪酸增加和不饱和脂肪酸减少的量也相对较大;贮藏初期超高压处理组与未处理组样品TBARS值都没有显著变化,而随贮藏期延长,超高压处理组样品TBARS值比未处理组略有上升。超高压处理能够有效延长低温烟熏火腿的货架期,不会或较小引起游离脂肪酸、贮藏期内色泽和脂肪氧化指标的显著变化,能够较好的保持产品原有游离脂肪酸组成及口感品质。     

不同贮藏温度西式火腿切片品质变化规律研究

为研究市场上低温火腿切片的质量和安全性,对不同贮藏温度的真空包装火腿切片的品质变化进行动态跟踪,分析产品在0～4、7～11℃条件下,其感官品质、菌落总数、大肠菌群、色泽、pH值、水分含量、保水性和质构特性的动态变化。结果表明：两种产品品质变化的规律大致相同。0～4℃贮藏比7～11℃产品的菌落总数增长相对缓慢,pH值、水分含量,保水性和质构特性的变化更加稳定。低温更有利于保持产品品质的稳定性和安全性,延长肉品的货架期。     

Protective cultures inhibit growth of Listeria monocytogenes and Escherichia coli O157:H7 in cooked, sliced, vacuum- and gas-packaged meat

Contamination of cooked meat products with Listeria monocytogenes poses a constant threat to the meat industry. The aim of this study was therefore to investigate the use of indigenous lactic acid bacteria (LAB) as protective cultures in cooked meat products. Cooked, sliced, vacuum- or gas-packaged ham and servelat sausage from nine meat factories in Norway were inoculated with 10(3) cfu/g of a mixture of three rifampicin resistant (rif-mutant) strains of L. monocytogenes and stored at 8 degrees C for four weeks. Growth of L. monocytogenes and indigenous lactic acid flora was followed throughout the storage period. LAB were isolated from samples where L. monocytogenes failed to grow. Five different strains growing well at 3 degrees C. pH 6.2, with 3% NaCl, and producing moderate amounts of acid were selected for challenge experiments with the rif-resistant strains of L. monocytogenes. a nalidixic acid/streptomycin sulphate-resistant strain of Escherichia coli O157:H7 and a mixture of three rif-resistant strains of Yersinia enterocolitica O:3. All five LAB strains inhibited growth of both L. monocytogenes and E. coli O157:H7. No inhibition of Y. enterocolitica O:3 was observed. A professional taste panel evaluated cooked, sliced, vacuum-packaged ham inoculated with each of the five test strains after storage for 21 days at 8 degrees C. All samples had acceptable sensory properties. The five LAB strains hybridised to a 23S rRNA oligonucleotide probe specific for Lactobacillus sakei. These indigenous LAB may be used as protective cultures to inhibit growth of L. monocytogenes and E. coli O157:H7 in cooked meat products.     

Thermal inactivation D- and z-values of multidrug-resistant and non-multidrug-resistant Salmonella serotypes and survival in ground beef exposed to consumer-style cooking

There has been speculation that multidrug-resistant (MDR) strains are generated by subtherapeutic antibiotic use in food animals and that such strains result in increased resistance to lethality by food processes such as heat and irradiation. The objective of this study was to evaluate the heat resistance of 20 strains, namely an MDR and a non-multidrug-resistant (NMDR) strain of each of 10 Salmonella serotypes isolated from cattle or cattle environments. MDR and NMDR Salmonella serotypes studied included Montevideo, Typhimurium, Anatum, Muenster, Newport, Mbandaka, Dublin, Reading, Agona, and Give. For phase I, stationary-phase cultures of the strains were aliquoted into sterile capillary tubes and immersed in a temperature-controlled water bath at 55, 60, 65, and 70 degrees C for appropriate times. Survivor curves were plotted for each temperature, and a best-fit linear regression was derived for each temperature. D-values (decimal reduction times) and z-values (changes in temperature required to change the D-values) were calculated for each strain. Although there was no overall significant difference in the heat resistance of MDR and NMDR serotypes, NMDR serotypes generally appeared to have slightly higher heat resistance than NMDR serotypes, especially at 55 and 60 degrees C. The highest relative heat resistance (highest z-values) was exhibited by Salmonella Anatum. Notably, the relative heat resistance of NMDR Salmonella Agona was similar to that of NMDR Salmonella Anatum and had the highest D-values at all four temperatures. For phase II, three serotypes (regardless of resistance profile) with the highest relative heat resistance and their drug-resistant counterparts were selected for thermal inactivation in ground beef patties cooked to endpoint temperatures. Salmonella Agona was able to survive in ground beef cooked to an internal temperature of 71 degrees C. Results of these studies suggest drug resistance does not affect the heat resistance of Salmonella and that serotype or strain is an important consideration in risk assessment of the pathogen with regard to survival at cooking temperatures.     

Survival of antibiotic resistant and antibiotic sensitive strains of E. coli O157 and E. coli O26 in food matrices

Escherichia coli O157:H7 or E. coli O26, which were AS (antibiotic sensitive), AR (laboratory created antibiotic resistant mutants), or naturally MAR (multi-antibiotic resistant), were inoculated into laboratory media, yoghurt or orange juice and their growth/survival monitored during enrichment at 37 degrees C or storage at 4 degrees C. The strains were also inoculated into minced beef and their thermal inactivation (D-values) examined at 55 degrees C, with and without a prior heat shock at 48 degrees C. The growth kinetics (lag phases, growth rates) of the VTEC (verocytotoxigenic E. coli), incubated over 24 h at 37 degrees C in laboratory media, were similar regardless of the presence or absence of antibiotic resistance. In yoghurt and orange juice, E. coli O157:H7 MAR died off significantly faster (P<0.05) than any of other VTEC strains examined. E. coli O157:H7 MAR was also found to be significantly more heat sensitive (P<0.05) than the other VTEC strains tested. The reasons for the observed differences in survival of the different VTEC strains and the link between antibiotic resistance and survival in VTEC organisms are discussed.