Comparison of Five Media for the Isolation of Coliform Organisms from Dehydrated and Deep Frozen Foods

S ummary . A significantly higher number of coliform and faecal coliform organisms were isolated using lactose-glutamic acid medium than by using lauryl sulphate-tryptone, lactose, brilliant green-bile or EE broths. With brilliant green-bile and lauryl sulphate-tryptone broth, coliforms/Enterobacteriaceae were isolated less often than with the others. For the detection of faecal coliforms, EE broth proved less good than all other media. All 5 media showed more coliforms/Enterobacteriaceae after 48 h than after 24 h at 30°. Few false positive results were obtained using any of these media.     

Optimum Membrane Structures for Growth of Coliform and Fecal Coliform Organisms

The purpose of this study was to determine the optimum membrane filter structure and characteristics for recovery of coliform organisms. Additionally, other factors such as sterilization method and membrane composition were examined. Fecal coliform growth tests with varied samples indicated that the most critical factor in recovery was surface pore morphology and not other factors previously suspected. Fecal coliform counts showed a dramatic increase, with increasing surface opening sizes. Membrane structures with surface openings large enough to surround the entrapped bacteria are required for optimum growth of fecal coliform organisms. Maximum fecal coliform recoveries are obtained using membranes composed of mixed esters of cellulose exhibiting a surface opening diameter of 2.4 μm and a retention pore size of 0.7 μm.     

Radiometric Method for the Detection of Coliform Organisms in Water

A new radiometric method for the detection of coliform bacteria in water has been described. The method is based on the release of ¹⁴CO₂ from [¹⁴C]lactose by bacteria suspended in growth medium and incubated at 37 C. The evolved ¹⁴CO₂ is trapped by hyamine hydroxide and counted in a liquid scintillation spectrometer. The method permits the detection of 1 to 10 organisms within 6 h of incubation. Coliform bacteria suspended in water for several days recover from starvation and may be quantitated by the proposed method. Bacteria from water samples may also be concentrated by filtration through membrane filters and detected by the radiometric assay.     

Comparative Analysis of Modeling Techniques for Coliform Organisms in Streams

The use of models for predicting changes in water quality parameters is currently considered an integral part of river basin management. The application of modeling techniques to coliform organisms is in its infancy due to the complexities involved and the lack of definitive information on coliform populations in natural environments. The purpose of this study was to make a comparative analysis of the available models for coliform organisms in order to improve on the state of the art of this subject. The available coliform models may be classified into deterministic or statistical types. In this study, six different models, three of each type, were selected for analysis and were applied to coliform data available on the Leaf River. Results of comparing the models indicated that a deterministic model was best suited for total coliform and a statistical model was best suited for fecal coliform. Ultimate selection of a model for coliform organisms is dependent not only on the accuracy of the model but on ease of implementation. Current technology would probably dictate the use of a deterministic model because of the lack of a complete data base on which to base statistical models.     

应用不同方法检测食品中大肠菌群结果的比较

评价检测食品中大肠菌群不同方法。比较国家标准、行业标准和显色培养基检测方法检测大肠菌群结果的差别。国家标准和行业标准检测结果基本一致,但有差异,应用显色培养基检测大肠菌群优于目前使用的国家标准和出口食品检验行业标准方法。检测食品中大肠菌群,显色培养基检测方法快速、灵敏、特异。     

Comparison of Fecal Coliform Agar and Violet Red Bile Lactose Agar for Fecal Coliform Enumeration in Foods

A 24-h direct plating method for fecal coliform enumeration with a resuscitation step (preincubation for 2 h at 37 ± 1°C and transfer to 44 ± 1°C for 22 h) using fecal coliform agar (FCA) was compared with the 24-h standardized violet red bile lactose agar (VRBL) method. FCA and VRBL have equivalent specificities and sensitivities, except for lactose-positive non-fecal coliforms such as Hafnia alvei, which could form typical colonies on FCA and VRBL. Recovery of cold-stressed Escherichia coli in mashed potatoes on FCA was about 1 log unit lower than that with VRBL. When the FCA method was compared with standard VRBL for enumeration of fecal coliforms, based on counting carried out on 170 different food samples, results were not significantly different (P > 0.05). Based on 203 typical identified colonies selected as found on VRBL and FCA, the latter medium appears to allow the enumeration of more true fecal coliforms and has higher performance in certain ways (specificity, sensitivity, and negative and positive predictive values) than VRBL. Most colonies clearly identified on both media were E. coli and H. alvei, a non-fecal coliform. Therefore, the replacement of fecal coliform enumeration by E. coli enumeration to estimate food sanitary quality should be recommended.     

转基因产品免疫学检测技术研究进展

目前转基因产品的检测主要有基于DNA的检测方法以及基于蛋白质的免疫学检测方法,简述了免疫学检测方法中的免疫印迹法(Western blotting)、酶联免疫吸附试验(ELISA)和试纸条法的原理、特点、应用例子以及存在问题。对一些新的方法如免疫PCR法、生物传感器法也略作介绍。     

Coliform Behavior in Frozen Foods: I. Rapid Test for the Recovery of Escherichia coli from Frozen Foods

An assortment of 496 samples of frozen foods consisting of fish or marine products, variety types, and cream pie desserts were subjected to four parallel examinations for the recovery of Escherichia coli. The test procedures consisted of two low-temperature (35 C) and two high-temperature (44 C) presumptive tests, followed by an E C confirmatory test at 45.5 C. Of all the test methods examined, a single Lauryl Sulfate Tryptose (LST) presumptive test at 44 C gave best E. coli recovery (425). This recovery compared favorably with the lengthier Association of Official Analytical Chemists test with which only 420 E. coli cells were recovered. The LST (44 C) test saves much time, since it renders a follow-up 48-hr confirmatory test unnecessary. Moreover, since 96% of all the E. coli are recovered within 24 hr by LST (44 C), it is essentially a 24-hr test. The results of this study also confirmed earlier findings, in that it is possible to describe a specific coliform bacteriological test method by simple reproducible productivity ratios. E. coli recovery dilution data and coliform group behavior were also examined.     

The Practical Significance in the Microbiological Examination of Cold Stored Foods of the Allegedly Low Heat Resistance among Psychrotrophic Organisms

Summary: The significance in food microbiology of the observation of Stapert, Sokolski & Northam (1962), that some bacteria occurring in water have such low heat resistances that they would be affected by the warm agar used for pouring plates, was tested. Forty-two samples of 6 different foods stored for 7 days at 3° to enrich their psychrotrophic microflora, were examined using the same medium, but in thin poured plates and on spread plates, respectively, incubated at 14°. The logarithmic average count of the foods was c. 10⁸/g and included about 60% Gram-negative rods and 30% cocci. In 90% of the samples no differences were observed between the results from the two methods of counting. The slightly higher spread plate counts in the remaining instances could be accounted for by the increased disruption of bacterial agglomerations always observed in this procedure. Hence there is no reason to query the results of poured plate counts in general. Where it is yet probably safer to use spread plates, rigorous asepsis in pouring and storing should be observed to prevent the development of colonies of contaminants too small to be detected when the plates are inoculated, but leading to erroneous counts after incubation.     

Statistical Analysis of the Recovery of Coliform Organisms on Gelman and Millipore Membrane Filters

The recovery of coliform organisms on Gelman and Millipore membranes was analyzed by using both a model I (which assumes no error in the x variable) and model II (which allows errors in both the variables) regression analysis. The two models afford estimates of the slope which agree within their 95% confidence limits. Using equations derived in this paper, the model II confidence limits on the intercept are obtained. This range does not include the model I intercept limits, thereby demonstrating the differences between results from an incorrect (model I) and correct (model II) approach. In addition, fecal coliform show no differences in response to the two membranes, whereas total coliform exhibit higher recoveries on Gelman membranes.     

Examination of Prepared Foods in Plastic Packages for Clostridium botulinum

The incidence of Clostridium botulinum organisms was determined in a variety of plastic-packaged "vulnerable" foods (food requiring little or no heating prior to consumption). A total of 113 foods were examined by use of an enrichment recovery procedure followed by toxin testing in animals. Results of the survey indicate that the incidence of C. botulinum organisms in these vulnerable foods is extremely low. The ability of inoculated food products to support growth and toxigenesis of C. botulinum type E was then tested. The 64 packaged foods were inoculated with type E spores and incubated anaerobically at 30 C for 11 days. A slurry of each food was prepared, smears for fluorescent-antibody testing were made, and animal tests were performed for toxin. If the animal tests were negative, enrichment cultures were prepared from the slurry and incubated at 30 C. On direct examination of the slurries for toxin, only samples of turkey roll and soybean cake supported growth and toxigenesis by C. botulinum type E. However, the enrichment culture method was able to induce growth and toxin production in 60 of the remaining 62 samples.     

Mathematical-Ecological Aspects of the Examination for Enterobacteriaceae of Foods Processed for Safety

S ummary : When the minimum proportion of Enterobacteriaceae to salmonellae in dried foods processed for safety is of the order of 10³, as the authors' surveys show them to be, testing 1 or 2 × 1 g aliquots for Enterobacteriaceae and allowing no positives confers the same degree of consumers' protection as examining 60 × 25 g amounts directly for salmonellae and accepting the consignment only when no positives are detected.     

Inhibition of Shigella flexneri by the Normal Intestinal Flora II. Mechanisms of Inhibition by Coliform Organisms

Of 15 strains of coliform bacteria, all isolated from human feces, 14 inhibited the growth of Shigella flexneri in mixed culture. In every case, when inhibition occurred, exponential growth of Shigella was interrupted in the mixed culture and the organisms entered into either a stationary or a death phase. None of the test coliform strains produced colicines active against Shigella. An analysis of mixed-culture environments at the time Shigella inhibition occurred revealed that the inhibition was not due to nutrient depletion nor to the development of adverse pH or oxidation-reduction potentials in themselves. In mixed cultures, the coliform strains produced formic and acetic acids in concentrations that inhibited Shigella growth. With one exception, the coliform strains also greatly reduced the culture medium. In average concentrations produced, the formic and acetic acids exerted a bactericidal effect on Shigella under the reduced conditions found in mixed cultures. The acids were only moderately toxic for the coliform strains under the same conditions. Results indicate that volatile acid production and concomitant reduction of the medium are the mechanisms by which coliform bacteria inhibit Shigella growth in mixed cultures.     

Simple, Reliable Cold Tray for the Recovery and Examination of Thermosensitive Organisms

A simple chamber is described for the isolation and handling of thermosensitive bacteria on board ship and in conventional research or classroom laboratories.     

Antimicrobial Actions of Degraded and Native Chitosan against Spoilage Organisms in Laboratory Media and Foods

The objective of this study was to determine whether chitosan (poly-β-1,4-glucosamine) and hydrolysates of chitosan can be used as novel preservatives in foods. Chitosan was hydrolyzed by using oxidative-reductive degradation, crude papaya latex, and lysozyme. Mild hydrolysis of chitosan resulted in improved microbial inactivation in saline and greater inhibition of growth of several spoilage yeasts in laboratory media, but highly degraded products of chitosan exhibited no antimicrobial activity. In pasteurized apple-elderflower juice stored at 7°C, addition of 0.3 g of chitosan per liter eliminated yeasts entirely for the duration of the experiment (13 days), while the total counts and the lactic acid bacterial counts increased at a slower rate than they increased in the control. Addition of 0.3 or 1.0 g of chitosan per kg had no effect on the microbial flora of houmous, a chickpea dip; in the presence of 5.0 g of chitosan per kg, bacterial growth but not yeast growth was substantially reduced compared with growth in control dip stored at 7°C for 6 days. Improved antimicrobial potency of chitosan hydrolysates like that observed in the saline and laboratory medium experiments was not observed in juice and dip experiments. We concluded that native chitosan has potential for use as a preservative in certain types of food but that the increase in antimicrobial activity that occurs following partial hydrolysis is too small to justify the extra processing involved.     

Detection and Enumeration of Fecal Indicator Organisms in Frozen Sea Foods: II. Enterococci

Consistently high recoveries of enterococci as compared to the low numbers of coliforms obtained from the same samples of frozen sea foods are indirect evidence that enterococci are better indicators of contamination in such foods.

The use of azide dextrose broth, modified by the incorporation of bromthymol blue, and of ethyl violet azide broth as presumptive and confirmation tests, respectively, were found to be highly specific for the detection and enumeration of enterococci in these samples. Tetrazolium agar medium, when used as a third step after the confirmation test, provides a reliable differentiation of Streplococcus faecalis types from other group D streptococci. A simple procedure is described for further identification of S. faecalis varieties and other enterococcal species.

Incidence of biotypes within certain species is noted and relationships of these subgroups to the organisms described by other workers is discussed.

The striking resistance of all group D streptococci to dihydrostreptomycin and polymyxin B seems to offer promise for evolving a new selective medium for these organisms.