草鱼鱼腩中热杀索丝菌生长预测模型的建立与验证

以鱼腩中热杀索丝菌为研究对象,根据恒定温度下的活菌数以及利用Gompertz模型得到的预测值,绘制不同温度下热杀索丝菌的实际和预测生长曲线,可以直观判断曲线重合度较好。预测模型方程判定系数R2均在0.99以上,显著性方差均小于0.0001,结果表明Gompertz模型对不同温度下热杀索丝菌的生长动态拟合良好。利用平方根模型描述温度与最大比生长速率和延滞期的关系,构建了热杀索丝菌的二级模型。通过计算准确因子和偏差因子对一级模型进行验证,计算结果均在1左右,利用F统计量的方法对二级模型进行验证,得到P值小于0.05。结果表明形成的生长预测一级模型和二级模型能够很好的描述0～15℃下热杀索丝菌在鱼腩上的生长情况。     

鲜切结球莴苣中单增李斯特菌生长预测模型的建立

为建立不同温度下鲜切结球莴苣中单增李斯特菌生长模型,将单增李斯特菌接种到鲜切结球莴苣表面,并于不同温度下贮藏,获得其在4、8、16、24和32℃下的生长数据,选用Gompertz模型进行拟合,建立初级生长模型。在此基础上建立二级模型研究温度对初级模型中单增李斯特菌生长动力学参数的影响,并进行数学检验。结果表明,对最大比生长速率和延滞时间建立平方根模型,结果呈良好的线性关系,相关系数R2分别为0.977 2和0.984 7,所建立的预测模型能很好地描述不同温度下单增李斯特菌的生长动态。     

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。     

冷鲜猪肉馅中热杀索丝菌生长预测模型的建立与验证

以市售托盘装冷鲜猪肉馅为研究对象,测定其中热杀索丝菌分别在0、5、10、15、20℃条件下的数量变化情况。运用统计学软件SAS9.1拟合热杀索丝菌在不同温度下的生长动力模型,采用偏差因子和准确因子进行验证,偏差因子和准确因子的值都在1左右,判定系数R2的值接近1,表明Gompertz模型能很好拟合热杀索丝菌在不同温度下的生长;利用平方根模型描述温度与最大比生长速率和延滞期的关系,得到热杀索丝菌生长的二级模型,方差分析结果显示P<0.01,表明模型能有效预测0~20℃温度区域内冷鲜猪肉馅中的热杀索丝菌的生长。     

草鱼鱼糜中沙门氏菌生长预测模型的建立

为建立鱼糜中沙门氏菌生长预测模型,选用新鲜草鱼鱼糜和鼠伤寒沙门氏菌作为研究对象,比较了4、10、20、28、37℃条件下鱼糜中鼠伤寒沙门氏菌的生长情况,分别采用Huang模型,Baranyi模型和修正的Gompertz模型进行拟合,建立鱼糜中鼠伤寒沙门氏菌一级生长动力学模型。并用平方根模型方程描述温度与比生长速率和延滞期的关系,得到鼠伤寒沙门氏菌生长二级模型。使用判定系数R2,准确因子(Af),偏差因子(Bf)和均方误差(MSE)对一级和二级模型可靠性进行评价,结果表明修正的Gompertz模型更适合于描述4~37℃条件下鱼糜中鼠伤寒沙门氏菌的生长变化,二级平方根模型可用于描述鱼糜中鼠伤寒沙门氏菌的生长参数,能够为鱼糜中沙门氏菌的监测提供一定的参考依据。     

冷鲜猪腩肉假单胞菌生长动力学模型的分析研究

以托盘包装冷鲜猪腩肉为研究对象,研究假单胞菌的生长变化与模型的构建和验证。结果表明:Gompertz方程拟合获得假单胞菌生长预测一级模型,相关系数R2值均大于0.99,总的平均偏差因子和准确因子均在1左右;利用平方根模型研究温度对一级模型动力学参数的影响,得到温度与最大比生长速率和延滞期方程分别为姨U=0.063×[T-(-12.79)]和姨1/LPD=0.086×[T-(-12.67)],其中T∈[0,20],R2分别为0.972和0.851,F统计量检验表明方程具有显著性。结论:建立的生长预测一级和二级模型能够很好的描述0℃~20℃下假单胞菌在冷鲜猪腩肉基质上的生长。     

单增李斯特菌在生食鱼片中生长模型的建立

为研究生食鱼片中单增李斯特菌的生长规律,将单增李斯特菌接种到经冷杀菌后的3 种生食鱼片（三文鱼片、金枪鱼片、鲷鱼片）中,分别置于4、8、15、25、35 ℃环境下培养,间隔适当时间取出计数。用5 种常用的一级模型（Gompertz模型、Baranyi模型、Logistic模型、Richards模型和MMF模型）对实验数据进行拟合,通过比较相关系数R2和均方误差（mean square error,MSE）,确定最适一级模型为Gompertz模型。建立单增李斯特菌生长动力学参数（最大比生长速率μm和迟滞期λ）关于温度、pH值和水分活度的二级平方根扩展模型,并应用相关系数R2、偏差值Bf和准确值Af进行验证。结果表明,构建的二级模型能够很好地描述生食鱼片中单增李斯特菌的生长情况。     

盐水鸭中单核细胞增生李斯特菌生长预测模型的建立

为研究盐水鸭中单核细胞增生李斯特菌(Listeriamonocytogenes,Lm)的生长规律,通过测定4、10、16、25℃条件下的生长数据,选用4种常用的一级模型(Gompertz、Logistic、Richards及MMF模型)对数据进行拟合,比较各模型决定系数R^2和均方误差(MSE),确定最适一级模型,根据一级模型得到的最大比生长速率(μmax)和迟滞期(λ)建立与温度相关的二级模型。结果表明:Gompertz模型拟合的生长曲线R^2均达到0.99以上,为最适一级模型,在25℃条件下,Lm经0.78 h后即进入对数期,从4℃提高到10℃时,生长速率从0.02 1g(cfu/g)·h^-1增至0.05 1g(cfu/g)·h^-1,说明温度对盐水鸭中Lm的生长影响较大。选用Ratkowsky平方根模型建立的温度与μmax关系的二级模型R^2为0.98,偏差因子(Bf)、准确因子(Af)分别为0.99、1.01,二次多项式模型建立的温度与λ关系的R^2为0.99,Bf、Af分别为1.01、1.08,表明所建两种模型均能较好地描述盐水鸭中Lm的生长情况。本研究建立的生长模型可为监控盐水鸭的食品安全和风险评估提供参考。     

气调包装冷却牛肉特定腐败菌生长模型的建立

以气调包装冷却牛肉为研究对象,研究储藏在0、5、10℃温度条件下气调包装冷却牛肉中特定腐败菌(假单胞菌)的生长变化规律。通过Matlab7.0 软件拟合不同温度条件下假单胞菌的生长情况,得到假单胞菌生长的Gompertz 模型参数;利用响应面方程建立假单胞菌生长的二级模型;由平方根方程建立了温度对假单胞菌生长动力学影响的数学模型。进而验证0～10℃低温条件下假单胞菌的生长动力学模型。结果表明：建立的Gompertz 方程拟合相关系数均在0.99 以上,二级模型偏差度和准确度均在0.90～1.05 之间,说明建立的一级和二级模型能够真实有效地预测0～10℃条件下气调包装冷却牛肉中假单胞菌的生长情况。     

鲜切紫薯中酵母菌和乳酸菌货架期模型的构建

用Gompertz模型对不同温度条件下真空包装鲜切紫薯中酵母菌和乳酸菌的生长曲线进行一级模型拟合,并通过计算准确因子（Af）和偏差因子（Bf）验证的生长模型的准确性;利用平方根模型和Arrhenius模型构建酵母菌和乳酸菌的二级模型,比较两种模型的优劣性;依据真空紫薯感官评价判定紫薯的腐败限控量,对其货架期预测模型进行构建和验证。结果表明,Gompertz模型能较好地拟合不同温度条件下2 种菌的生长情况（R2在0.9左右）,数学检验参数Af、Bf接近1.0,均在接受范围;通过不同二级模型动力学参数的比较,2 种菌均为用平方根模型拟合二级模型较优;紫薯选用酵母菌作为指示菌预测货架期较好,其腐败限控量为6.64 （lg（CFU/g））,选定4 ℃条件构建并验证货架期预测模型可靠性,实测值为7.0 d,预测值为7.737 d,预测效果良好。     

Effects of gas atmosphere, antimicrobial dip and temperature on the fate of Listeria innocua and Listeria monocytogenes on minimally processed lettuce

The growth and survival of inoculated strains of Listeria innocua and L. monocytogenes on minimally processed lettuce were studied. The effects of package atmospheres (lettuce sealed within packages after flushing with 100% N₂ or without flushing with N₂, lettuce sealed within perforated packages), antimicrobial dips (100 p.p.m. chlorine solution for 5 min, 1% citric acid solution for 5 min) and storage temperatures (3°C and 8°C) were investigated. Populations of L. innocua and L. monocytogenes on undipped lettuce stored at 3°C gradually decreased (by 1–1.5 log cycles) during a 14 day storage period. By contrast counts on lettuce stored at 8°C did not change significantly ( P > 0.05). Flushing packages of lettuce with 100% N₂ followed by storage at 8°C resulted in a significant increase ( P < 0.05, by 2–3 log cycles) in L. innocua and L. monocytogenes counts during storage. L. innocua , strain NCTC 11288, behaviour was similar to that of L. monocytogenes (strains ATCC, 19114 and NCTC 11994) under these storage temperatures and atmospheres. Using L. innocua as a model for L. monocytogenes , it was found that dipping lettuce in a chlorine or citric acid solution followed by storage at 8°C resulted in a significant increase ( P < 0.05, by 2 log cycles) in L. innocua populations compared with undipped samples. It is concluded that N₂ flushing or use of antimicrobial dips combined with storage at 8°C, both enhanced the survival and growth of Listeria populations on shredded lettuce.     

Growth of Listeria monocytogenes on iceberg lettuce and solid media

The growth of pathogenic bacterium Listeria monocytogenes on fresh-cut iceberg lettuce under constant temperatures was modelled in order to investigate microbial safety during distribution of this vegetable. We examined the effects of several incubation temperatures, ranging from 5 to 25 degrees C, on bacterial growth. These data were fitted to the Baranyi model and the curves showed a high correlation coefficient at all temperature (R2 > 0.95). In addition, the native bacterial flora of the lettuce did not affect the growth rate of L. monocytogenes regardless of incubation temperature. However, the lag time was reduced at a ratio of native bacteria to inoculated L. monocytogenes (100:1) at low incubation temperatures (5 and 10 degrees C). Furthermore, the maximum population density (MPD) was increased at a low ratio of native to inoculated L. monocytogenes (1:1) at all incubation temperatures. These results were compared with the previous work published by [Buchanan, R.L., Stahl, H.G., Whiting, R.C., 1989. Effects and interactions of temperature, pH, atmosphere, sodium chloride, and sodium nitrite on the growth of Listeria monocytogenes. J. Food Prot. 52, 844-851] that is being developed at the US Department of Agriculture (USDA) Agricultural Research Service's Pathogen Modelling Program (PMP). The broth-based Buchanan model for L. monocytogenes was found to markedly deviate from the observed data. In order to investigate this discrepancy, we examined the effects of medium environment and nutrient content on L. monocytogenes growth using tryptic soy agar plates (TSAP) and agar plates (AP) containing 1.7% sucrose. The inoculated bacteria on both TSAP and AP showed slower growth rates than that predicted by the PMP. The MPD of bacteria grown on TSAP was similar to the PMP model ( approximately 9 log10 CFU/ml or plate (circle of diameter of 90 mm)) regardless of the incubation temperature. By contrast, the MPD observed on AP was approximately 4 log10 CFU lower than that observed on TSAP or predicted by the PMP. Both the growth rate and the MPD of L. monocytogenes on AP were similar to those on lettuce. These results suggest that the solid medium and poor nutrient content inhibited the growth of L. monocytogenes on lettuce. The growth rates of the inoculated L. monocytogenes on all media were described using Ratkowsky's simple square root model.     

Modeling and predicting the simultaneous growth of Listeria monocytogenes and natural flora in minced tuna

The growth kinetics of Listeria monocytogenes and natural flora (NF) in minced tuna from 2 to 30 °C were examined, and a simultaneous growth model was developed. The inhibiting effect of the NF on the growth of L. monocytogenes was examined by inoculating different levels of NF isolated from the minced tuna. The kinetic data were fitted to the Baranyi model and estimated the growth parameters such as specific growth rate (μ(max)), maximum population density (N(max)), and lag time. The temperature and inoculated NF dependency on the μ(max) of L. monocytogenes and NF were described by modified Ratkowsky's square-root model. As the initial NF level increased, the slopes of the square-root models were decreased for both L. monocytogenes and NF. The N(max) of L. monocytogenes was described as a function of temperature and inoculated NF level. Simultaneous growth prediction of L. monocytogenes and NF under constant temperature conditions was examined by using the differential equations based on the Baranyi model with the effect of interspecies competition substituted into the developed μ(max) and N(max) models. The root mean square errors between the model prediction and the observation for L. monocytogenes and NF were 0.42 and 0.34, respectively. Predictive simulation under fluctuating temperature conditions also demonstrated a high accuracy of simultaneous prediction for both L. monocytogenes and NF, representing the root mean square errors of 0.19 and 0.34, respectively. These results illustrate that the developed model permits accurate estimation of the behavior of L. monocytogenes in minced tuna under real temperature history until consumption.     

Efficacy of disinfectants to reduce Listeria monocytogenes on precut iceberg lettuce

The efficacy of water, chlorinated water (100 ppm), peracetic acid solution (0.05%), and commercial citric acid-based produce wash (0.25%) to reduce the population of Listeria monocytogenes on precut lettuce was tested. Samples were inoculated with a mixture of equal amounts of five L. monocytogenes strains at a level of 4.7 log CFU/g, and analyzed on the day of washing and after 3 and 6 days of storage at 6 degrees C. Sanitizer reduced the number of L. monocytogenes at maximum 1.7 log CFU/g and number of L. monocytogenes reached the inoculation level during 6 days of storage. Thus, disinfectants do not eliminate L. monocytogenes on precut lettuce and cannot be solely relied on in producing precut lettuce safely. The inoculated L. monocytogenes strains were recovered at different rates after 6 days of storage; one of these strains was not recovered at all. Thus, strain-specific differences exist in the ability of L. monocytogenes to survive the washing treatments of the lettuce.     

Prediction of pathogen growth on iceberg lettuce under real temperature history during distribution from farm to table

The growth of pathogenic bacteria Escherichia coli O157:H7, Salmonella spp., and Listeria monocytogenes on iceberg lettuce under constant and fluctuating temperatures was modelled in order to estimate the microbial safety of this vegetable during distribution from the farm to the table. Firstly, we examined pathogen growth on lettuce at constant temperatures, ranging from 5 to 25 degrees C, and then we obtained the growth kinetic parameters (lag time, maximum growth rate (micro(max)), and maximum population density (MPD)) using the Baranyi primary growth model. The parameters were similar to those predicted by the pathogen modelling program (PMP), with the exception of MPD. The MPD of each pathogen on lettuce was 2-4 log(10) CFU/g lower than that predicted by PMP. Furthermore, the MPD of pathogens decreased with decreasing temperature. The relationship between mu(max) and temperature was linear in accordance with Ratkowsky secondary model as was the relationship between the MPD and temperature. Predictions of pathogen growth under fluctuating temperature used the Baranyi primary microbial growth model along with the Ratkowsky secondary model and MPD equation. The fluctuating temperature profile used in this study was the real temperature history measured during distribution from the field at harvesting to the retail store. Overall predictions for each pathogen agreed well with observed viable counts in most cases. The bias and root mean square error (RMSE) of the prediction were small. The prediction in which mu(max) was based on PMP showed a trend of overestimation relative to prediction based on lettuce. However, the prediction concerning E. coli O157:H7 and Salmonella spp. on lettuce greatly overestimated growth in the case of a temperature history starting relatively high, such as 25 degrees C for 5 h. In contrast, the overall prediction of L. monocytogenes under the same circumstances agreed with the observed data.     

Reduction of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes with electrolyzed oxidizing water on inoculated hass avocados (Persea americana var. Hass)

This study was intended to evaluate the bactericidal effect of electrolyzed oxidizing water (EOW) and chlorinated water on populations of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes inoculated on avocados (Persea americana var. Hass). In the first experiment, inoculated avocados were treated with a water wash applied by spraying tap water containing 1 mg/liter free chlorine for 15 s (WW); WW treatment and then spraying sodium hypochlorite in water containing 75 mg/liter free chlorine for 15 s (Cl75); WW treatment and then spraying alkaline EOW for 30 s (AkEW) and then spraying acid EOW (AcEW) for 15 s; and spraying AkEW and then AcEW. In another experiment, the inoculated avocados were treated by spraying AkEW and then AcEW for 15, 30, 60, or 90 s. All three pathogen populations were lowered between 3.6 and 3.8 log cycles after WW treatment. The application of Cl75 did not produce any further reduction in counts, whereas AkEW and then AcEW treatment resulted in significantly lower bacterial counts for L. monocytogenes and E. coli O157:H7 but not for Salmonella. Treatments with AkEW and then AcEW produced a significant decrease in L. monocytogenes, Salmonella, and E. coli O157:H7 populations, with estimated log reductions of 3.9 to 5.2, 5.1 to 5.9, and 4.2 to 4.9 log CFU/cm2, respectively. Spraying AcEW for more than 15 s did not produce any further decrease in counts of Salmonella or E. coli O157:H7, whereas L. monocytogenes counts were significantly lower after spraying AcEW for 60 s. Applying AkEW and then AcEW for 15 or 30 s seems to be an effective alternative to reduce bacterial pathogens on avocado surfaces.     

Comparison of the attachment of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, and Pseudomonas fluorescens to lettuce leaves

Attachment of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella Typhimurium, and Pseudomonas fluorescens on iceberg lettuce was evaluated by plate count and confocal scanning laser microscopy (CSLM). Attachment of each microorganism (approximately 10(8) CFU/ml) on the surface and the cut edge of lettuce leaves was determined. E. coli O157:H7 and L. monocytogenes attached preferentially to cut edges, while P. fluorescens attached preferentially to the intact surfaces. Differences in attachment at the two sites were greatest with L. monocytogenes. Salmonella Typhimurium attached equally to the two sites. At the surface, P. fluorescens attached in greatest number, followed by E. coli O157:H7, L. monocytogenes, and Salmonella Typhimurium. Attached microorganisms on lettuce were stained with fluorescein isothiocyanate and visualized by CSLM. Images at the surface and the cut edge of lettuce confirmed the plate count data. In addition, microcolony formation by P. fluorescens was observed on the lettuce surface. Some cells of each microorganism at the cut edge were located within the lettuce tissues, indicating that penetration occurred from the cut edge surface. The results of this study indicate that different species of microorganisms attach differently to lettuce structures, and CSLM can be successfully used to detect these differences.     

Determination of the growth limits and kinetic behavior of Listeria monocytogenes in a sliced cooked cured meat product: validation of the predictive growth model under constant and dynamic temperature storage conditions

To describe the growth limits of Listeria monocytogenes NCTC10527 in a sliced vacuum-packaged cooked cured meat product, the binary logistic regression model was used to develop an equation to determine the probability of growth or no growth of L. monocytogenes as a function of temperature (from 0 to 10 degrees C) and water activity (from 0.88 to 0.98). Two inoculum concentrations were used (10 and 10(4) CFU g(-1)), and the growth limits for the two inocula were different. The kinetic behavior of L. monocytogenes as a function of temperature (4, 8, 12, and 16 degrees C) on the same meat product at the lower concentration (10 CFU g(-1)) was also studied. The Baranyi model appeared to fit the overall experimental data better than did the modified Gompertz and the modified logistic models. Maximum specific growth rate (micromax), lag phase duration (LPD), and maximum cell concentration (Nmax) derived from the primary model were modeled using the square root function (micromax and LPD) and a second order polynomial (Nmax) (secondary models). The selection of the best model (primary or secondary) was based on some statistical indices (the root mean square error of residuals of the model, the regression coefficient, the F test, the goodness of fit, and the bias and accuracy factor). The developed kinetic behavior model was validated under constant and dynamic temperature storage conditions. This prediction of L. monocytogenes growth provides useful information for improving meat safety and can be used for in-depth inspection of quality assurance systems in the meat industry.     

Effects of the indigenous microflora of minimally processed lettuce on the survival and growth of Listeria innocua

Interactions between the natural background microflora of shredded lettuce and Listeria innocua (in lieu of Listeria monocytogenes ) were studied. The effect of increasing the initial size of indigenous populations (from 10³ to 10⁶–10⁷ CFU g⁻¹) was tested for its ability to reduce L. innocua growth on shredded lettuce. Co-culture experiments were performed in model media, where bacterial isolates from the indigenous microflora were tested for possible inhibitory effects. Varying the size of the indigenous populations had no effect on L. innocua survival or growth. However, interactions with individual species and mixed populations from lettuce did affect the survival and growth of L. innocua in model media. In general, mixed populations diminished L. innocua growth. In the undiluted lettuce medium, the various species tested individually either reduced or did not affect the growth of L. innocua . However, when the medium was diluted, some species extended the survival of L. innocua . Competition between the indigenous microflora and L. innocua resided mostly with the Enterobacter spp. and not with the pseudomonads. Enterobacter cloacae was particularly effective in reducing L. innocua growth. Lactic acid bacteria also reduced L. innocua growth in undiluted media. It is concluded that interactions with the natural background microflora may play an important role in determining the dynamics of Listeria populations on shredded lettuce.     

Assessment of control measures to achieve a food safety objective of less than 100 CFU of Listeria monocytogenes per gram at the point of consumption for fresh precut iceberg lettuce

The important new concept of the food safety objective (FSO) offers a strategy to translate public health risk into a definable goal such as a specified maximum frequency or concentration of a hazardous agent in a food at the time of consumption that is deemed to provide an appropriate level of health protection. For the foodborne pathogen Listeria monocytogenes, there is a proposed FSO of < 100 CFU/g in ready-to-eat (RTE) products at the time of consumption. Fresh precut iceberg lettuce is one of these RTE products. In this study, we worked with a commercial manufacturer to evaluate the effectiveness of two antimicrobial washing agents (sodium hypochlorite and a mixture of hydrogen peroxide and peroxyacetic acid) against L. monocytogenes under simulated fresh precut washing conditions and evaluated the growth potential of this pathogen on lettuce packaged in a gas-permeable film and stored at 4 or 8 degrees C for 14 days. We used the results of this experiment to demonstrate how the commercial manufacturer could meet the FSO for L. monocytogenes in fresh precut lettuce through the application of performance, process, and microbiological criteria.