Rapid dye reduction tests for the determination of microbiological quality of meat

Rapid dye reduction tests have been developed to determine the quality of meat. Three chemical indicators, resazurin and two tetrazolium compounds, were used to correlate the microbial numbers and reduction times in meat samples. Twenty-five surface samples from sheep carcasses were subjected to each reduction test. Total viable counts given were obtained at 37°C. Resazurin reduction time was 90–120 min when the bacterial counts ranged from 1.5×10⁶ to 7.7×10⁶/cm². Samples showing bacterial counts between 1.5×10⁶ and 6.0×10⁶/cm² reduced tetrazolium (NBT) in 360–390 min whereas samples containing bacterial counts of 2.1×10⁶/cm² took 420–450 min to reduce iodophenyl nitrophenyl tetrazolium (INT) dye. Regression equations relating the number of organisms per cm² and reduction time were applied to predict the microbiological quality of meat samples from reduction time data. Among the three dyes, resazurin gave the lowest reduction time.     

An interlaboratory comparison of methods used to assess antioxidant potentials

Many analytical methods are used to measure the antioxidative activity of substances yet little is known about the comparability of the test results between laboratories. After an initial evaluation of a broad range of methods conducted by one laboratory, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, the trolox equivalent antioxidant capacity (TEAC) assay, the lipid assay (or 2,2'-azobis(2-aminepropane) (ABAP) assay) and the thiobarbituric acid (TBA) assay were selected to be evaluated in the interlaboratory study. The antioxidative potentials of trolox, tocopherol, lipochroman-6, ascorbic acid, 4-methyl-brenzcatechin, and/or 3,5-di-tert-butyl-4-hydroxytoluene (BHT) were assessed using each of the methods. These methods were then evaluated in respect of their reproducibility and classification properties. Based on the results of this study, the DPPH assay followed by the TEAC assay yielded the best results based on reproducibility and sensitivity both within one laboratory and between laboratories. The results of the interlaboratory study were then compared with the single center results obtained from the commercially available photochemolumiescence (PCL) kit. To assess the transferability of chemical data to biological systems, they were also compared with the single center results obtained using the cell-based Dichlorodihydrofluoresceine (DCFH) assay.     

Evaluation and validation of two microbiological tests for screening antibiotic residues in honey according to the European guideline for the validation of screening methods

Two microbiological kits based on Bacillus stearothermophilus (Eclipse 50® and Premi®Test) have been evaluated and validated according to the European guideline for the validation of screening methods (January 2010) and in relation to the concentrations recommended by the EU-RL in 2007. Both tests are robust, a fast method and easy to implement. Both tests are applicable to a very large variety of honeys from different floral and geographical origins (rosemary, lavender, scrub, heath, alder, forest, lemon, acacia, chestnut, raspberry, mountain and flowers) as well as honey of different colours (from blank honey to brown honey, including yellow and orange honey). A satisfactory false-positive rate of 5% was obtained for the Eclipse 50® test. The observed detection capabilities CCβ of the Eclipse 50® kit were: chlortetracycline (>75?µg?kg^?1), oxytetracycline (≤200?µg?kg^?1), tetracycline (>100?µg?kg^?1), cloxacillin (≤40?µg?kg^?1), tylosin (≤200?µg?kg^?1), desmycosin (>400?µg?kg^?1), sulfadiazine (≤300?µg?kg^?1), sulfadimethoxine (≤250?µg?kg^?1), sulfamerazine (>300?µg?kg^?1), sulfamethazine (>1000?µg?kg^?1), sulfamethizole (>75?µg?kg^?1), sulfamethoxazole (≤25?µg?kg^?1), sulfanilamide (?1000?µg?kg^?1), sulfaquinoxaline (>75?µg?kg^?1), sulfathiazole (≤250?µg?kg^?1) and lincomycin (>1500?µg?kg^?1). These levels were all higher than the recommended concentrations where they exist. Due to its lack of sensitivity, it cannot be recommended for reliable routine use. The observed CCβ of the Premi®Test kit were: chlortetracycline (10?µg?kg^?1), oxytetracycline (>10?µg?kg^?1), tetracycline (≤10?µg?kg^?1), cloxacillin (≤5?µg?kg^?1), tylosin (≤10?µg?kg^?1), desmycosin (≤15?µg?kg^?1), sulfadiazine (≤25?µg?kg^?1), sulfadimethoxine (≤25?µg?kg^?1), sulfamerazine (≤25?µg?kg^?1), sulfamethazine (≤25?µg?kg^?1), sulfamethizole (≤25?µg?kg^?1), sulfamethoxazole (≤10?µg?kg^?1), sulfanilamide (≤25?µg?kg^?1), sulfaquinoxaline (≤10?µg?kg^?1), sulfathiazole (25?µg?kg^?1) and lincomycin (≤25?µg?kg^?1). The Premi®Test kit could be recommended for reliable use in routine control due to its low detection capabilities (except for aminoglycosides), but the disadvantage is a high false-positive rate of 14%.     

Experimental comparison of excision and swabbing microbiological sampling methods for carcasses

Bovine sides, ovine carcasses, and porcine carcasses were individually inoculated by dipping in various suspensions of a marker organism (Escherichia coli K-12 or Pseudomonas fluorescens), alone or in combination with two meat-derived bacterial strains, and were sampled by two standard methods: cotton wet-dry swabbing and excision. The samples were examined for bacterial counts on plate count agar (PCA plate counts) and on violet red brilliant green agar (VRBGA plate counts) by standard International Organization for Standardization methods. Average bacterial recoveries by swabbing, expressed as a percentage of the appropriate recoveries achieved by excision, varied widely (2 to 100%). Several factors that potentially contributed to relatively low and highly variable bacterial recoveries obtained by swabbing were investigated in separate experiments. Neither the difference in size of the swabbed area (10, 50, or 100 cm2 on beef carcasses) nor the difference in time of swabbing (20 or 60 min after inoculation of pig carcasses) had a significant effect on the swabbing recoveries of the marker organism used. In an experiment with swabs preinoculated with the marker organism and then used for carcass swabbing, on average, 12% of total bacterial load was transferred inversely (i.e., from the swab to the carcass during the standard swabbing procedure). In another experiment, on average, 14% of total bacterial load was not released from the swab into the diluent during standard swab homogenization. Use of custom-made swabs with abrasive butts, around which metal pieces of pan scourers were wound, markedly increased PCA plate count recoveries from noninoculated lamb carcasses at commercial abattoirs compared with cotton swabs. In spite of the observed inferiority of the cotton wet-dry swabbing method compared with the excision method for bacterial recovery, the former is clearly preferred by the meat industry because it does not damage the carcass. Therefore, further large-scale evaluation of the two carcass sampling methods has been undertaken under commercial conditions and reported separately.     

Sampling methods for microbiological analysis of red meat and poultry carcasses

Microbiological analysis of carcasses at slaughterhouses is required in the European Union for evaluating the hygienic performance of carcass production processes as required for effective hazard analysis critical control point implementation. The European Union microbial performance standards refer exclusively to the excision method, even though swabbing using the wet/dry technique is also permitted when correlation between both destructive and nondestructive methods can be established. For practical and economic reasons, the swab technique is the most extensively used carcass surface-sampling method. The main characteristics, advantages, and limitations of the common excision and swabbing methods are described here.     

Analysing collaborative trials for qualitative microbiological methods: accordance and concordance

In qualitative (detection) food microbiology, the usual measures of repeatability and reproducibility are inapplicable. For such studies, we introduce two new measures: accordance for within laboratory agreement and concordance for between laboratory agreement, and discuss their properties. These measures are based on the probability of finding the same test results for identical test materials within and between laboratories, respectively. The concordance odds ratio is introduced to present their relationship. A method to test whether accordance differs from concordance is discussed.     

关于三种实验室间比对数据统计分析方法的研究

目前国内实验室间比对常采用CNAS—GL02《能力验证结果的统计处理和能力评价指南》中推荐的稳健(Robust)统计法对数据进行统计分析评价,GB/T 6379.2—2004《测量方法与结果的准确度(正确度与精密度)第2部分确定标准测量方法重复性与再现性的基本方法》中推荐采用格拉布斯检验法(Grubbs)和科克伦(Co...     

An evaluation of methods for inoculating Listeria monocytogenes into butter for microbiological experimentation

The behaviour of Listeria monocytogenes Scott A was assessed when inoculated into butter using six different methods. The butter was made using a laboratory-scale butter churn to give consistency in the butter ingredients. No increase in count was seen using the 'stab' or 'contact' methods of butter inoculation. No inoculation method was found to be ideal for every inoculation experiment that may be required. However, it was concluded that the 'mix' and 'bead' methods were most appropriate if the 'cream' inoculation method could not be used.     

食品检验中的菌种保藏方法

综述了食品检验中常用的菌种保藏方法,包括定期移植保藏法、液体石蜡保藏法、载体保藏法、低温保藏法和冷冻干燥保藏法,旨在为食源性致病菌菌种的保藏提供参考。     

Petrifilm--up-to-date tests for microbiological control of food products, raw materials and environmental objects

The principle of action, advantages and different types of Petrifilm as alternative test to the classical microbiological analysis are considered. Petrifilm is up-to-date high technology test systems for the rapid quantitative microbiological control in foodstuff of sanitary-significant and pathogenic microorganisms--yeast and moulds, coliforms, Enterobacteriaceae, Staphylococcus, Listeria spp. and Lactobacilli spp. Automatic testing of colonies on Petrifilm with use of the Petrifilm-Reader allows to reduce time for colony counting, to exclude errors of personnel and to document results of the analysis.     

Measurement uncertainty of the EU methods for microbiological examination of red meat

Three parallel trials were made of EU methods proposed for the microbiological examination of red meat using two analysts in each of seven laboratories within the UK. The methods involved determination of aerobic colony count (ACC) and Enterobacteriaceae colony count (ECC) using simulated methods and a freeze-dried standardised culture preparation. Trial A was based on a simulated swab test, Trial B a simulated meat excision test and Trial C was a reference test on reconstituted inoculum. Statistical analysis (ANOVA) was carried out before and after rejection of outlying data. Expanded uncertainty values (relative standard deviation x2) for repeatability and reproducibility, based on the log10 cfu/ml, on the ACC ranged from +/-2.1% to +/-2.7% and from +/-5.5% to +/-10.5%, respectively, depending upon the test procedure. Similarly for the ECC, expanded uncertainty estimates for repeatability and reproducibility ranged from +/-4.6% to +/-16.9% and from +/-21.6% to +/-23.5%, respectively. The results are discussed in relation to the potential application of the methods.     

Changes in vegetable microbiological quality introduced by processing methods

The main objective of this research was to investigate the effect in microbial quality of vegetables, because of the chain of buyers and sellers involved in the collection, processing and selling of vegetables, from the primary production sector up to the consumer level (from farm to table). Two processing plants and six hotels were selected and 240 vegetables, 30 vegetable containers, 18 water samples used for sanitation purposes and samples of 18 personnel's hands, were microbiologically analysed. Based on actual results and processing plants auditing, we conclude that initial vegetable microbial quality is critical. The storage, separation and packaging processes, cleaning procedures and retention time at processing plants can influence vegetable microbial quality. Buyers–sellers involved between vegetable production in primary sector and final consumer can offer a dramatic decrease in vegetable quality. The use of ISO 9001:2000 can improve product quality through techniques such as product traceability and resources management.     

显齿蛇葡萄提取物抗菌作用的研究

为推进梨酒减硫增质进程,促进天然抗氧化剂在梨酒中的应用。本文通过单独使用二氢杨梅素（Dihydromyricetin,DMY）或与SO₂联合使用酿造库尔勒香梨酒,分别测定各组梨酒自由基清除率、总酚含量、总黄酮含量、酒体色度及风味物质,研究了DMY对梨酒抗氧化活性以及风味物质的影响。结果表明,各组梨酒基础理化指标无显著性差异（P>0.05）,发酵均能正常完成;H4组（SO₂ 30 mg/L、DMY 100 mg/L）抗氧化活性最佳,DPPH自由基清除率为70.08%,ABTS⁺自由基清除率达到95.89%;单独使用DMY（浓度为150或200 mg/L）或与SO₂联合使用时,均能促进酒体总酚及总黄酮的生成（P<0.05）。此外,DMY对梨酒挥发性风味物质影响较大,DMY浓度越高,越有益于主体香气物质的生成;DMY和SO₂联合使用亦能提高挥发性物质的种类和总量,其中H3组（SO₂ 30 mg/L、DMY 75 mg/L）风味物质种类达到24种,总含量高达11846.14 μg/L,乙酸乙酯和辛酸乙酯含量相比CK组分别增加了107.61、420.27 μg/L。综合来看,H3组总酚含量更高、颜色更加稳定,酒体香气更加浓郁、层次感愈加分明,感官品评高达85.7分 。 DMY与SO₂的混合使用在抑制褐变、促进总酚方面综合效果最佳,适量DMY可增加改善香梨酒香气成分,提高其品质。     

国外禽畜胴体非破坏性微生物样品采集方法分析

屠宰生产线上禽畜胴体微生物样品采集方法分可破坏性采样和非破坏性采样。本文收集并分析了国际标准化组织(International Standardization Organization,ISO)、美国农业部、新西兰食品安全局以及澳大利亚检验检疫局关于禽畜胴体非破坏性微生物样品采集的方法,其包括湿-干拭子法(wet and dry swab method)、海绵拭子法(sponge sampling method)和纱布敷料法(gauze tampon method)、胴体冲洗采样法,不同屠宰工艺不同微生物目标其采样时机、采样位点、采样频率和微生物限量不尽相同,以期为我国禽畜产品加工生产和安全监控提供采样技术参考。     

Suitability of rapid microbiological methods for the hygienic management of spray drier plant

A study was initiated to find the most suitable rapid microbiological method to monitor the hygienic performance and facilitate the hygienic management of a single-site spray drier/blending/packing operation. The advantages of employing a rapid method and the criteria and methods used to evaluate commercially available instruments are listed. A brief review of the DEFT, Bactec, Lumac, Bactobridge, Bactometer and Malthus instruments is given, from which we conclude that it is the impedimetric/conductivity instruments that show the greatest potential for the future. We selected the Malthus range of microbial growth analysers as the most suitable for our requirements.     

Redox potential measurement as a rapid method for microbiological testing and its validation for coliform determination

The redox potential is one of the most complex indicators of the physiological state of microbial cultures and its measurement could be a useful tool for the qualitative and quantitative determination of the microbial contamination. During the bacterial growth, the redox potential of the medium decreases. The shape of the redox potential curve is characteristic on the type of microorganism, and the rate of the change (dE/dt) is proportional to the living cell concentration. Defining the time required to reach a significant change in redox potential as Time to Detection (TTD), similarly to the impedimetric measurements, a strict linear correlation could be established between the TTD and the logarithm of the initial concentration of microorganisms. On the base of this calibration curve, the determination of living cell concentration could be simplified. For the experiments, a computer-controlled multi-channel measuring system and software was developed by the authors. The redox potential measurement method was tested and validated for the determination of coliform bacteria. The results have proved the high efficiency and reliability of the new method.     

微生物法测定乳粉中叶酸的不确定度评定

目的 评定微生物法测定乳粉中叶酸的不确定度.方法 按照GB 5009.211—2014《食品安全国家标准食品中叶酸的测定》规定的微生物法对婴儿配方乳粉中叶酸含量进行测定,根据CNAS-CL01-G003:2019要求,依据GB/Z 22553—2010和JJF 1059.1—2012,分析测定过程中影响检验结果的因素主...     

Inhibition of bacterial growth in thawed cake and rapid evaluation methods for microbiological quality

The inhibition of bacterial growth in thawed cake kept under refrigeration and rapid evaluation methods for microbiological quality of the cake were investigated. The effects of the freezing temperature and the addition of ethanol or emulsifier on bacterial numbers in a cake model after storage for 72 hr at 10 degrees C following the thawing process were also studied. Bacterial growth in the cake model was inhibited by the additives under various freezing conditions. In addition, rapid evaluation methods for estimating bacterial numbers in the cake model after incubation for 72 hr at 10 degrees C were studied. High correlations were found between bacterial numbers in the cake model incubated for 24 hr at 20 degrees C and for 6 hr at 35 degrees C with tryptic soy broth and that of the cake model incubated for 72 hr at 10 degrees C. This result indicated that rapid evaluation by incubation for 24 hr at 20 degrees C or for 6 hr at 35 degrees C with tryptic soy broth can be used to predict the bacterial numbers in a cake model after incubation for 72 hr at 10 degrees C. Furthermore, the ATP-bioluminescence method was applied to shorten the testing time, because culture on an agar medium was not necessary.