Rapid method for prediction of Escherichia coli numbers using an electronic sensor array and an artificial neural network

An electronic sensor array with 12 nonspecific metal oxide sensors was evaluated for its ability to monitor volatile compounds in super broth alone and in super broth inoculated with Escherichia coli (ATCC 25922) at 37 degrees C for 2 to 12 h. Using discriminant function analysis, it was possible to differentiate super broth alone from that containing E. coli when cell numbers were 10(5) CFU or more. There was a good agreement between the volatile profiles from the electronic sensor array and a gas chromatography-mass spectrometer method. The potential to predict the number of E. coli and the concentration of specific metabolic compounds was investigated using an artificial neural network (ANN). The artificial neural network was composed of an input layer, one hidden layer, and an output layer, with a hyperbolic tangent sigmoidal transfer function in the hidden layer and a linear transfer function in the output layer. Good prediction was found as measured by a regression coefficient (R2 = 0.999) between actual and predicted data.     

Detection of Escherichia coli in packaged alfalfa sprouts with an electronic nose and an artificial neural network

A rapid method for the detection of Escherichia coli (ATCC 25922) in packaged alfalfa sprouts was developed. Volatile compounds from the headspace of packaged alfalfa sprouts, inoculated with E. coli and incubated at 10 degrees C for 1, 2, and 3 days, were collected and analyzed. Uninoculated sprouts were used as control samples. An electronic nose with 12 metal oxide electronic sensors was used to monitor changes in the composition of the gas phase of the package headspace with respect to volatile metabolites produced by E. coli. The electronic nose was able to differentiate between samples with and without E. coli. To predict the number of E. coli in packaged alfalfa sprouts, an artificial neural network was used, which included an input layer, a hidden layer, and an output layer, with a hyperbolic tangent sigmoidal transfer function in the hidden layer and a linear transfer function in the output layer. The network was shown to be capable of correlating voltametric responses with the number of E. coli. A good prediction was possible, as measured by a regression coefficient (R2 = 0.903) between the actual and predicted data. In conjunction with the artificial neural network, the electronic nose proved to have the ability to detect E. coli in packaged alfalfa sprouts.     

近红外光谱协同BP神经网络的泰国茉莉香米掺伪含量快速鉴定

利用近红外光谱协同BP神经网络算法,对泰国茉莉香米及其掺伪样品的近红外光谱进行多元散射校正预处理,挑选出48个特征波长;以特征波长的吸光度为BP神经网络输入层神经元,以样品中泰国茉莉香米的含量为输出层神经元,获得BP神经网络算法的最优结构模型,即为单层隐含层、隐含层神经元数7、隐含层传递函数logsig、输出层传递函数...     

Effect of Soybean Volatile Compounds on Aspergillus flavus Growth and Aflatoxin Production

ABSTRACT:  Soybean homogenates produced volatile compounds upon exposure to lipase. These induced volatiles were identified by SPME. Seventeen volatile compounds identified by SPME were chosen for determination of their ability to inhibit Aspergillus flavus growth and aflatoxin B₁ (AFB1) production in a solid media assay. These volatiles included aldehydes, alcohols, ketones, and furans. Of the tested compounds, the aldehydes showed the greatest inhibition of fungal growth and AFB1 production. These compounds inhibited up to 100% of the observed growth and AFB1 production as compared to the controls. The greatest activity by the aldehydes to disrupt growth was ranked as follows: 2,4 hexadienal > benzaldehyde > 2-octenal > ( E )-2-heptenal > octanal > ( E )-2-hexenal > nonanal > hexanal. The greatest activity by the aldehydes to reduce AFB1 was ranked as follows: ( E )-2-hexenal > 2,4 hexadienal > ( E )-2-heptenal > hexanal > nonanal. ( E )-2-hexenal and ( E )-2-heptenal were tested further in an A. flavus -inoculated corn kernel assay. Both compounds prevented colonization by A. flavus and eliminated AFB1 production when exposed to compound volumes < 10 μL as also shown in the solid media assay. The results suggest that soybeans react to lipase by production of potent antifungal volatiles.     

顶空固相微萃取和同时蒸馏萃取法分析猪肉汤的挥发性成分

采用顶空固相微萃取法(HS-SPME)和同时蒸馏萃取法(SDE)提取清炖猪肉汤中挥发性成分,结合气相色谱-质谱联用技术(GC-MS)进行分析,结果共检测出77种挥发性化合物,其中烃类和醛类最多,均有22种,其次是醇类17种和酸类6种,另外酯类、酮类和杂环类分别有4、3、3种。HS-SPME法和SDE法分别鉴定出42种和49种挥发性组分。两种提取方法相比较,采用SPME法提取到醛类的种类明显多于SDE法,而烃类、醇类和酯类的种类明显少于SDE法。SDE法适合对高沸点、低挥发性物质的分离;SPME法具有快速简便、不使用溶剂和样品检测非破坏性等优点,适合易挥发性化合物的检出。只有将两种方法结合起来,才能得到对产品的挥发性物质的综合评价。     

GC-MS based metabolomics for rapid simultaneous detection of Escherichia coli O157:H7, Salmonella Typhimurium, Salmonella Muenchen, and Salmonella Hartford in ground beef and chicken

A metabolomic-based method for rapid detection of Escherichia coli O157:H7, Salmonella Hartford, Salmonella Typhimurium, and Salmonella Muenchen in nonselective media was developed. All pathogenic bacteria were grown in tryptic soy broth (TSB) at 37 °C followed by metabolite quantification at 2-h intervals for 24 h. Results were compared with the metabolite profiles similarly obtained with E. coli K12, Pseudomonas aeruginosa, Staphylococcus aureus, Saccharomyces cereviseae, and Aspergillus oryzae grown individually or as a cocktail under the same conditions. Principal component analysis (PCAS) discriminated pathogenic microorganisms grown in TSB. Metabolites responsible of PCAS classification were dextrose, cadaverine, the aminoacids L-histidine, glycine, and L-tyrosine, as well as the volatiles 1-octanol, 1-propanol, 1butanol, 2-ethyl-1-hexanol, and 2,5-dimethyl-pyrazine. Partial least square (PLS) models based on the overall metabolite profile of each bacteria were able to detect the presence of Escherichia coli O157:H7 and Salmonella spp. at levels of approximately 7 ± 2 CFU/25 g of ground beef and chicken within 18 h.     

Relationships of Overall Estery Aroma Character in Lagers with Volatile Headspace Congener Concentrations

In lager beers the intensity of “estery” aroma character is regarded as an important component of sensory quality, but its origins are somewhat uncertain. Overall “estery” aroma intensity was predicted from capillary gas chromatographic (GC) data following solid phase micro extraction (SPME) of headspaces. Estery character was scored in 23 commercial lagers using rankrating, allowing assessors (13) constant access to a range of appropriate standards. From univariate data analysis, all assessors behaved similarly and lagers fell into three significantly different groups: low (1), high (1) and intermediate (21). The quantification of 36 flavour volatiles by SPME of headspaces was reproducible and principal component analysis explained 91% total variance. Multiple linear regression could utilise only a restricted (26) set of flavour volatiles, whereas partial least square regression, that considered all flavour components, showed significant differences and improved prediction. However, an artificial neural network that could compensate for non‐linearities and interactions in ester perception gave the most robust prediction at R² = 0.88.     

Attachment of Escherichia coli O157:H7 grown in tryptic soy broth and nutrient broth to apple and lettuce surfaces as related to cell hydrophobicity, surface charge, and capsule production

This study investigated the effect of growth in tryptic soy broth (TSB) and nutrient broth (NB) on the ability Escherichia coli O157:H7 to attach to lettuce and apple surfaces. In addition, cell surface hydrophobicity, charge and capsule production were determined on cells grown in these media. Cells grown in NB attached less to lettuce and apple surfaces than did those grown in TSB. TSB, but not NB, supported capsule production by E. coli O157:H7. Cells grown in TSB were more hydrophilic than those grown in NB. No difference was found in the electrokinetic properties of cells grown in these media. Electrostatic and hydrophobic interactions and surface proteins did not appear to play an important role in the attachment of E. coli O157:H7 to these surfaces. Of the factors studied, only capsule production was associated with attachment ability.     

Comparison of methods for detection and isolation of cold- and freeze-stressed Escherichia coli O157:H7 in raw ground beef

A comparison was made of the relative efficiencies of three enrichment media, RapidChek Escherichia coli O157:H7 enrichment broth (REB), R&F broth (RFB), and modified E. coli broth containing novobiocin (mEC+n), and four selective plating media for detection of cold- and freeze-stressed E. coli O157:H7 in raw ground beef. Ground beef (25 g) was inoculated with E. coli O157:H7 at < or =0.5 and < or =2 CFU/g, and samples were then enriched immediately or were stored at 4 degrees C for 72 h or at -20 degrees C for 2 weeks and then enriched. After 8 or 20 h of enrichment, the cultures were plated onto R&F E. coli O157: H7 chromogenic plating medium, cefixime-tellurite sorbitol MacConkey agar, CHROMagar O157, and Rainbow agar O157 and tested using the RapidChek E. coli O157 lateral flow immunoassay and a multiplex PCR assay targeting the E. coli O157: H7 eae, stx1, and stx2 genes. Recovery of E. coli O157:H7 on the four agar media was 4.0 to 7.9 log CFU/ml with the REB enrichment, 1.4 to 7.4 log CFU/ml with RFB, 1.7 to 6.7 log CFU/ml with mEC+n incubated at 42 degrees C, and 1.3 to 3.3 log CFU/ml from mEC+n incubated at 35 degrees C. The percentages of positive ground beef samples containing nonstressed, cold-stressed, and freeze-stressed E. coli O157:H7 as obtained by plating, the immunoassay, and the PCR assay were 97, 88, and 97%, respectively, with REB, 92, 81, and 78%, respectively, with RFB, 97, 58, and 53%, respectively, with mEC+n incubated at 42 degrees C, and 22, 31, and 25%, respectively, with mEC+n incubated at 35 degrees C. Logistic regression analyses of the data indicated significant main effects of treatment, type of medium, enrichment time, inoculum concentration, and detection method. In particular, a positive result was 1.1 times more likely to occur after 20 h of enrichment than after 8 h, 25 times more likely with RFB and REB than with mEC+n at 35 degrees C, 3.7 times more likely with an initial inoculum of < or = 2.0 CFU/g than with < or = 0.5 CFU/g, 2.5 to 3 times more likely using freeze-stressed or nonstressed bacteria than with cold-stressed bacteria, and 2.5 times more likely by plating than by the immunoassay or the PCR assay. REB had better overall performance for enrichment of cold- and freeze-stressed E. coli O157:H7 present in ground beef than did the other media examined.     

Phagocytosis and serum susceptibility of Escherichia coil cultured in iron-deplete and iron-replete media

The susceptibility of Escherichia coli cultured in either iron-deplete or iron-replete media to phagocytosis by bovine neutrophils and the bactericidal activity of bovine serum was tested in vitro. Fourteen E. coli isolates from naturally occurring intramammary infections (IMI) were cultured overnight at 37 degrees C in iron-replete media and iron-deplete media. The iron-replete media were trypticase soy broth or a chemically defined medium. The iron-deplete media were either trypticase soy broth plus 0.2 mM alpha, alpha' dipyridyl and 1mM citrate, or the chemically defined medium plus 0.2 mM alpha, alpha' dipyridyl, and 1 mM citrate. Iron-replenished medium was the chemically defined iron-deplete medium plus 40 mM ferric citrate. Bacteria grown in iron-deplete media were less susceptible to phagocytosis compared with bacteria grown in iron-replete media. Replenishing the chemically defined iron-deplete medium with ferric citrate obliterated the decreased susceptibility to phagocytosis observed in iron-deplete media. The iron availability in media used to culture E. coli before assay did not affect the bactericidal action of either the classical pathway of complement or the antibody independent alternative pathway of complement in serum. The growth of bacteria in iron-deplete medium did not alter the expression of capsule compared with growth in iron-replete medium. Iron availability during culture of E. coli altered the susceptibility of isolates to phagocytosis by neutrophils, but had no effect on the susceptibility of isolates to the bactericidal activity of serum.     

A neuro-computing approach for modeling of residence time distribution (RTD) of carrot cubes in a vertical scraped surface heat exchanger (SSHE)

A neuro-computing approach was used for modeling two residence time distribution (RTD) functions — the time-specific (E-type distribution) and the cumulative particle concentration function (F-type distribution) — of carrot cubes in starch solutions in a vertical scraped surface heat exchanger (SSHE) of a pilot scale aseptic processing system. Experimental data obtained for E (t) and F(t) under various test conditions were used for both training and evaluation. Multi-layered artificial neural network (ANN) models with four input and two output neurons were trained. The network was optimized by the varying number of hidden layers, number of neurons in each hidden layer and learning runs, and a combination of learning rule and transfer functions, using a back-propagation algorithm. The trained ANN model was validated by a set of independent experimental data. The ANN models were also compared with conventional models developed based on multiple regression techniques. The results indicated that there was better agreement between experimental and ANN model predicted values for both E (t) and F (t) functions. The average modeling errors associated with ANN were 5.7 and 3.0%, respectively, for E(t) and F(t), while they were 15.5 and 12.3%, respectively, with the multiple regression models.     

红景天苷缓释微囊神经网络建模

本文在确定输入层和输出层单元、预处理网络数据、选择激活函数、选择训练方法的基础上,建立了红景苷缓释微囊制作参数与性能之间的神经网络模型,网络结构为5-12-3。该模型能较为精确的拟合输入的样本数据,其最大相对误差不超过4%,模型准确可信,可以代替真实试验,该模型的建立为工艺参数的优化打下基础。     

Analysis of brandy aroma by solid‐phase microextraction and liquid–liquid extraction

Headspace solid‐phase microextraction (SPME) and continuous liquid–liquid extraction (LLX) with Freon were used to extract and analyse aroma volatiles in brandy. In general, SPME using a non‐polar polydimethylsiloxane coating was more selective for esters and acids than was LLX. LLX using Freon 11 extracted the higher alcohols more efficiently than SPME. Relative differences in volatiles between brandies made from Vitis vinifera L cv Colombard and Vitis vinifera L cv Ugni blanc were observed, particularly for hexanol, 3‐methylbutylacetate, 3‐methylbutanol and 3‐methylbutyloctanoate. In addition, a combination of SPME with GC–olfactometry was used to provide more detailed information on sensory characteristics of varietal brandies. © 2000 Society of Chemical Industry     

Evaluation of Escherichia coli O157:H7 growth media for use in test-and-hold procedures for ground beef processing

Since the mid-1990s, the beef industry has used a process called test and hold, wherein beef trim and ground beef are tested to keep products contaminated with Escherichia coli O157:H7 out of commerce. Current O157:H7 detection methods rely on a threshold level of bacterial growth for detection, which is dependent on the growth medium used. Twelve media were examined for growth and doubling time: buffered peptone water (BPW), SOC (which contains tryptone, yeast extract, KCl, MgCl2, and glucose), buffered peptone water plus SOC (BPW-SOC), Bacto-NZYM, RapidChek E. coli O157:H7 medium, BioControl EHEC8 culture medium, Neogen Reveal for E. coli O157:H7--Eight Hour medium (Neogen Reveal 8), BAX System medium for E. coli O157:H7 (BAX) BAX System medium for E. coli O157:H7 MP (BAX-MP), modified E. coli broth, nutrient medium, and tryptic soy broth (TSB). All media were tested at 37 or 42 degrees C under static or shaking conditions. The eight media with the highest total CFU per milliliter and most rapid doubling times were BPW-SOC, NZYM, RapidChek, EHEC8, Neogen Reveal 8, BAX, BAX-MP, and TSB. The ability of these eight media to enrich E. coli O157:H7 in ground beef was further evaluated through time-course experiments using immunomagnetic separation. Of these media, TSB was the easiest to prepare, had a wide application base, and was the least expensive. In the test-and-hold process, the normal ratio of medium to product is 1:10. In this study, a 1:3 ratio worked as well as a 1:10 ratio. Processors using test-and-hold procedures could use 1 liter of TSB to enrich for E. coli O157:H7 in a 375-g sample instead of the usual 3.375 liters, thus saving reagents, time, and labor while maintaining accuracy.     

SDE and SPME Analysis of Flavor Compounds in Jin Xuan Oolong Tea

Simultaneous distillation‐extraction (SDE) and solid phase micro extraction (SPME) are procedures used for the isolation of flavor compounds in foods. The purpose of this study was to optimize SDE conditions (solvent and time) and to compare SDE with SPME for the isolation of flavor compounds in Jin Xuan oolong tea using GC‐MS and GC‐O. The concentration of volatile compounds isolated with diethyl ether was higher (P < 0.05) than for dichloromethane and concentration was higher at 40 min (P < 0.05) than 20 or 60 min extractions. For SDE, 128 volatiles were identified using GC‐MS and 45 aroma active compounds using GC‐O. Trans‐nerolidol was the most abundant compound in oolong tea. The number of volatiles identified using GC‐MS was lower in SPME than SDE. For SPME, 59 volatiles and 41 aroma active compounds were identified. The composition of the volatiles isolated by the 2 methods differed considerably but provided complementary information.     

Growth history influences starvation-induced expression of uspA, grpE, and rpoS and subsequent cryotolerance in Escherichia coli O157:H7

In this study, we investigated the effect of starvation on cryotolerance of Escherichia coli O157:H7 grown in tryptic soy broth (TSB) and Luria-Bertani broth (LB). Starved cells (cells suspended in water at 37 degrees C for 6 h) and control cells (cells in TSB or LB) were frozen at -18 degrees C for up to 240 h in their respective growth media. The E. coli grown in TSB showed a greater starvation effect (the difference in percent survival of starved and control cells) and cryotolerance. The starved E. coli grown in TSB showed a 30% increase in their ability to survive frozen storage for 24 h at -18 degrees C. The corresponding increase in survival for LB-grown E. coli was only 3.8%. Cryotolerance induced by starvation of TSB- and LB-grown E. coli was correlated with the expression of genes involved in general stress response pathways, such as uspA, grpE, and rpoS. The expression of uspA, grpE, and rpoS was quantified by measuring the green fluorescence generated from autofluorescent E. coli harboring puspA::gfp, pgrpE::gfp, and prpoS::gfp gene fusions. The results obtained in this study indicate that uspA, grpE, and rpoS were induced on starvation when E. coli was grown in TSB, and their expression correlated well with subsequent induction of cryotolerance developed at -18 degrees C. In contrast, cells grown in LB and subsequently exposed to starvation conditions showed no increase in expression of uspA, grpE, or rpoS, and, as expected, these cells did not exhibit increased cryotolerance at -18 degrees C. Knowledge of molecular mechanisms involved in cross-protection might make it possible to devise strategies to limit their effects and lead to ways to predict the survival of foodborne pathogens in stressful environments.     

Acquisition of antibiotic resistance plasmids by enterohemorrhagic Escherichia coli O157:H7 within rumen fluid

The emergence of antibiotic resistance among important foodborne pathogens like Escherichia coli O157:H7 has become an important issue with regard to food safety. In contrast to the case for Salmonella, antibiotic resistance has been slow in its development in E. coli O157:H7 despite the presence of mobile antibiotic resistance genes in other E. coli organisms that inhabit the same animal host. We set out to determine if rumen fluid influences the transfer of plasmid-mediated, antibiotic resistance to E. coli O157:H7. A commensal E. coli strain from a dairy cow was transformed with conjugative R plasmids and served as the donor in matings with naladixic acid-resistant E. coli O157:H7. R plasmids were transferred from the donor E. coli strain to E. coli O157:H7 in both Luria-Bertani (LB) broth and rumen fluid. R plasmids were transferred at a higher frequency to E. coli O157:H7 during 6 h of incubation in rumen fluid at rates comparable to those in LB broth, indicating that conditions in rumen fluid favor the transfer of the plasmids to E. coli O157. This finding suggests that the cow's rumen is a favorable environment for the genetic exchange of plasmids between microflora and resident E. coli O157:H7 in the bovine host.     

Impact of trehalose,sucrose and/or maltodextrin addition on aroma retention in freeze dried strawberry puree

Trehalose was studied as a drying aid to establish its impact on aroma retention in freeze dried strawberry puree. The evaluation was done by sensory analysis and headspace‐solid phase microextraction‐gas chromatography with a mass detector (HS‐SPME‐GC‐MS); results were compared with those obtained using sucrose and maltodextrin (MD) as drying aids. The carbohydrate used significantly modified the type and concentration of volatiles retained during freeze drying. HS‐SPME‐GC‐MS outcomes showed that the use of trehalose (alone or with MD) resulted in the product with the chromatographic profile most similar to fresh strawberry puree, being different from sucrose and MD. Meanwhile, sensory analysis showed a similarity with the aromatic profiles when using trehalose or sucrose, remaining both different from MD. This study proved that the use of trehalose as a drying aid can be beneficial on volatile aroma retention and that different combinations of organic volatiles can lead to similar sensory profiles.     

Predicting the drying kinetics of salted codfish (Gadus Morhua): semi‐empirical,diffusive and neural network models

This work aims to compare the accuracy of several drying modelling techniques namely semi‐empirical, diffusive and artificial neural network (ANN) models as applied to salted codfish (Gadus Morhua). To this end, sets of experimental data were collected to adjust parameters for the models. Modelling of codfish drying was performed by resorting to Page and Thompson semi‐empirical models and to a Fick diffusion law. The ANN employed a neural network multilayer ‘feed‐forward’, consisting of one input layer, with four neurons, one hidden layer, formed by five neurons and one output layer with a convergence criterion for training purposes. The simulations showed good results for the ANN (correlation coefficient between 0.987 and 0.999) and semi‐empirical models (correlation coefficient ranging from 0.992 to 0.997 for Page’s model, and from 0.993 to 0.996 for Thompson’s model), while improvements were required to obtain better predictions by the diffusion model (correlation coefficients ranged from 0.864 to 0.959).     

Efficacy of chromocult coliform agar for coliform and Escherichia coil detection in foods

Chromocult coliform agar (CCA) was compared with Petrifilm Escherichia coli count plate (PEC) for identifying coliforms and E. coli in a variety of meat products. Products examined included 45 raw beef samples, 12 sausage emulsion samples, 11 samples of meat-based ready-to-eat appetizers, and 8 pork trimming samples. Coliforms from CCA and PEC were confirmed by gassing in brilliant green lactose broth plus a positive reaction on purple broth agar plus lactose after incubation at 35 degrees C for 48 h. Lauryl sulfate tryptose plus methylumbelliferyl-beta-glucuronide and tryptophan broth were used to confirm E. coli from CCA and PEC with 48-h incubations at 35 and 42.5 degrees C, respectively. API 20E test strips were inoculated for final confirmation. The overall respective confirmation percentages (CFU/g) for the PEC and the CCA methods were 93.1 and 93.7% for coliforms and 99.8 and 98.1% for E. coli, although the CCA method yielded significantly (P < 0.001) higher mean CFU/g values for both coliforms and E. coli. Regression analyses of these data indicated a strong positive linear relationship existed between the two methods over a wide CFU/g range for both coliforms and E. coli. The respective correlation coefficients obtained for coliforms and E. coli of 0.89 and 0.86 indicate that the CCA method provides a reliable optional method for these determinations in meat products.