Detection Of Microbial Contamination Research Articles

Electronic Noses as Flexible Tools to Assess Food Quality and Safety: Should We Trust Them?

This paper presents three different applications of an electronic nose (EN) based on a metal oxide sensor array, in order to illustrate the broad spectrum of potential uses of the technique in food quality control. The following scenarios are considered: 1) the screening of a typical error that may occur during the processing of tomato pulp, which leads to sensory damage of the product; 2) the detection of microbial contamination by Alicyclobacillus spp. (ACB) affecting soft drinks; and 3) the proof of evidence of extra virgin olive oil fraudulently adulterated with hazelnut oil. In each case, the EN is able to identify the spoiled product by means of the alterations in the pattern of volatile compounds, reconstructed by principal component analysis of the sensor responses.

Analysis and detection of microbial contamination and biofilms in technical fluids and its impact on fluid management

Analysis and detection of microbial contamination and biofilms in technical fluids and its impact on fluid management

Solar disinfection of wastewater to reduce contamination of lettuce crops by Escherichia coli in reclaimed water irrigation

Low-cost disinfection methods to allow safe use of recycled wastewater for irrigation can have important beneficial implications in the developing world. This study aims to assess the efficiency of solar disinfection to reduce microbial contamination of lettuce crops when solar-treated wastewater effluents are used for irrigation. The irrigation study was designed as a complete experimental loop, including (i) the production of irrigation water through solar disinfection of real municipal wastewater treatment plant effluents (WWTPE), (ii) the watering of cultivated lettuce crops at the end of solar treatment, and (iii) the detection of microbial contamination on the irrigated crops 24 h after irrigation. Solar disinfection was performed using two types of reactors: (i) 20-L batch borosilicate glass reactors equipped with CPC to optimize solar irradiation, and (ii) 1.5-L PET bottles, i.e. the traditional SODIS recipients commonly used for disinfection of drinking water in developing communities. Both solar and H2O2-aided solar disinfection processes were tested during ≤5 h exposure of WWTPE, and Escherichia coli inactivation was analysed. A presence/absence detection method was developed to analyse lettuce leaves sampled 24 h after watering for the detection of E. coli. Results of inactivation assays show that solar disinfection processes can bring down bacterial concentrations of >103–104E. coli CFU mL−1 in real WWTPE to <2 CFU/mL (detection limit). The absence of E. coli on most lettuce samples after irrigation with solar-disinfected effluents (26 negative samples/28) confirmed an improved safety of irrigation practices due to solar treatment, while crops irrigated with raw WWTPE showed contamination.

A suitable and efficient procedure for the removal of decontaminating antibiotics from tissue allografts

The presence of residual antibiotics in tissue allografts after decontamination with antibiotic cocktails may result in widely documented adverse effects in predisposed subjects. Moreover, antibiotic residues may mask contaminating microorganisms, resulting in falsely negative sterility tests, with potential risk of post-surgical infections. The objective of the present study was to define a rinsing procedure capable of eliminating antibiotic residues from cardiovascular, bone and skin tissues after decontamination with BASE.128. Different washing patterns, employing BASE medium, were applied. The presence of antibiotic residues in tissue homogenates was assessed by agar diffusion test at different stages of tissue processing. To test whether antibiotic residues can result in falsely negative microbiological analysis, we induced a superficial tissue contamination with known inoculum concentration. By employing four different porcine tissues, we here report direct evidence that the presence of even limited amounts of antibiotics in decontaminated tissues interferes with sterility testing. This has implications in terms of increased risk of infections in allograft recipients. To minimize this risk, we developed a procedure for extensive removal of antibiotics from allografts, allowing for subsequent detection of microbial contaminations that may occur during transportation, storage or processing prior to allograft transplantation. Our study emphasizes the importance of validating all processes and analytical methods in tissue banking, in order to warrant tissue safety. This will minimize the risks of post-surgical infections as well as antibiotic-induced anaphylaxis in predisposed patients.

Identification of microbial volatile organic compounds (MVOCs) emitted from fungal isolates found on cinematographic film

We report the detection of microbial contamination and growth of cinematographic film utilising headspace solid phase micro-extraction coupled with Gas Chromatography–Mass Spectrometry. Microbial Volatile Organic Compounds (MVOCs) are produced only when the mould is actively growing on the cine film. Over 150 volatile compounds were detected from 16 fungal isolates, with over 40 being common to 2 or more isolates. It was found that 1-octen-3ol was produced from 13 of the isolates analysed, 3-octanone from 10 of the isolates and 3-octanol from 4 isolates. These three key chemical markers are indicative of viable fungal growth on cinematographic film precluding the need for traditional microbiology laboratory culture methods. Such an approach would prevent valuable historical footage from being discarded due to health and safety concerns regarding spore inhalation, and would enable safe handling.

Electronic Nose for Microbiological Quality Control of Food Products

Electronic noses (ENs) have recently emerged as valuable candidates in various areas of food quality control and traceability, including microbial contamination diagnosis. In this paper, the EN technology for microbiological screening of food products is reviewed. Four paradigmatic and diverse case studies are presented: (a)Alicyclobacillusspp. spoilage of fruit juices, (b) early detection of microbial contamination in processed tomatoes, (c) screening of fungal and fumonisin contamination of maize grains, and (d) fungal contamination on green coffee beans. Despite many successful results, the high intrinsic variability of food samples together with persisting limits of the sensor technology still impairs ENs trustful applications at the industrial scale. Both advantages and drawbacks of sensor technology in food quality control are discussed. Finally, recent trends and future directions are illustrated.

Validation of the BacT/ALERT®3D automated culture system for the detection of microbial contamination of epithelial cell culture medium

Living tissue engineering for regenerative therapy cannot withstand the usual pharmacopoeia methods of purification and terminal sterilization. Consequently, these products must be manufactured under aseptic conditions at microbiologically controlled environment facilities. This study was proposed to validate BacT/ALERT(®)3D automated culture system for microbiological control of epithelial cell culture medium (ECCM). Suspensions of the nine microorganisms recommended by the European Pharmacopoeia (Chap. 2.6.27: "Microbiological control of cellular products"), plus one species from oral mucosa and two negative controls with no microorganisms were prepared in ECCM. They were inoculated in FA (anaerobic) and SN (aerobic) culture bottles (Biomérieux, Lyon, France) and incubated in a BacT/ALERT(®)3D automated culture system. For each species, five sets of bottles were inoculated for reproducibility testing: one sample was incubated at the French Health Products Agency laboratory (reference) and the four others at Cell and Tissue Bank of Lyon, France. The specificity of the positive culture bottles was verified by Gram staining and then subcultured to identify the microorganism grown. The BacT/ALERT(®)3D system detected all the inoculated microorganisms in less than 2 days except Propionibacterium acnes which was detected in 3 days. In conclusion, this study demonstrates that the BacT/ALERT(®)3D system can detect both aerobic and anaerobic bacterial and fungal contamination of an epithelial cell culture medium consistent with the European Pharmacopoeia chapter 2.6.27 recommendations. It showed the specificity, sensitivity, and precision of the BacT/ALERT(®)3D method, since all the microorganisms seeded were detected in both sites and the uncontaminated medium ECCM remained negative at 7 days.

Production of Coturnix quail immunoglobulins Y (IgYs) against Vibrio parahaemolyticus and Vibrio vulnificus

Production of chicken immunoglobulins Y (IgYs) and their applications in prophylactic, therapeutic, detection of microbial contaminants and as a diagnostic tool has been widely studied with limited information from other avians. This study produced Coturnix quail (Coturnix coturnix japonica) egg yolk IgYs against Vibrio parahaemolyticus and Vibrio vulnificus. Formalin inactivated (FIVP, FIVV, mixed FI-VP/VV) and heat inactivated (HIVP, HIVV, mixed HI-VP/VV) Vibrio immunogens (109 CFU/mL) were intramuscularly immunized into quail through thigh muscles. Egg yolk IgY was purified by water dilution-ammonium sulfate precipitation method and the activity was determined by enzyme-linked immunosorbent assay (ELISA). Formalin inactivated immunogens induced high humoral immune response for both V. parahaemolyticus and V. vulnificus over heat inactivated immunogens. However, IgYs resulted from HIVP and FIVV immunogens, showed high specificity to V. parahaemolyticus and V. vulnificus respectively. Detection limits of the indirect ELISA using the produced IgYs were 105 CFU/mL for V. parahaemolyticus and 106 CFU/mL for V. vulnificus. The developed antibodies showed high binding affinity to their corresponding immunogens, very little cross reactivity to Staphylococcus aureus and not other bacteria strains (p<0.05), a phenomenon which was also observed in Western blot.

An Exometabolomics Approach to Monitoring Microbial Contamination in Microalgal Fermentation Processes by Using Metabolic Footprint Analysis

The early detection of microbial contamination is crucial to avoid process failure and costly delays in fermentation industries. However, traditional detection methods such as plate counting and microscopy are labor-intensive, insensitive, and time-consuming. Modern techniques that can detect microbial contamination rapidly and cost-effectively are therefore sought. In the present study, we propose gas chromatography-mass spectrometry (GC-MS)-based metabolic footprint analysis as a rapid and reliable method for the detection of microbial contamination in fermentation processes. Our metabolic footprint analysis detected statistically significant differences in metabolite profiles of axenic and contaminated batch cultures of microalgae as early as 3 h after contamination was introduced, while classical detection methods could detect contamination only after 24 h. The data were analyzed by discriminant function analysis and were validated by leave-one-out cross-validation. We obtained a 97% success rate in correctly classifying samples coming from contaminated or axenic cultures. Therefore, metabolic footprint analysis combined with discriminant function analysis presents a rapid and cost-effective approach to monitor microbial contamination in industrial fermentation processes.

Colorimetric bacteria sensing using a supramolecular enzyme-nanoparticle biosensor.

Rapid and sensitive detection of pathogens is a key requirement for both environmental and clinical settings. We report here a colorimetric enzyme-nanoparticle conjugate system for detection of microbial contamination. In this approach, cationic gold nanoparticles (NPs) featuring quaternary amine headgroups are electrostatically bound to an enzyme [β-galactosidase (β-Gal)], inhibiting enzyme activity. Analyte bacteria bind to the NP, which releases the β-Gal and restores its activity, providing an enzyme-amplified colorimetric readout of the binding event. Using this strategy, we have been able to quantify bacteria at concentrations of 1 × 10(2) bacteria/mL in solution and 1 × 10(4) bacteria/mL in a field-friendly test strip format.

Detection of microbial contaminants in mammalian cell cultures using a new fluorescence-based staining method

Microbial contamination of cell culture production processes is a current concern for biopharmaceutical industries. Traditional testing methods require several days to detect contamination and may advantageously be replaced by a rapid detection method. We developed a new method combining membrane filtration to microcolonies fluorescence staining method (MFSM) and compared it to epifluorescence microscopy. Both methods were used to detect bacteria in CHO cells cultures. The epifluorescence microscopy showed to be limited by filterability, media interference and nonrobustness issues, whereas MFSM enabled consistent detection of Bacillus cereus, Staphylococcus epidermidis and Propionibacterium acnes after, respectively, 8, 9 and 48 h of incubation. Thanks to the nondestructive feature of the MFSM, stained membranes could be reincubated on culture media to yield visible colonies used for identification. The new method described in this study showed its ability to detect microbial contaminants in cell culture samples with time-to-results from 2-5 times shorter than the traditional testing method. The MFSM can be used as monitoring tool for cell cultures to significantly shorten detection times of microbial contamination, while preserving the ability to identify the contaminants and their viability.

Fast detection of Candida albicans and/or bacteria in blood plasma by “sample-self-focusing” using capillary electrophoresis-laser-induced fluorescence

Detection of microbial contamination in blood plasma is critical and necessary in different medical and research fields. Most of the current standard procedures for the detection of bacteria and fungi can be time-consuming, for example, direct inoculation methods of microbial cultures in respective growth media can take a few days to several weeks. A fast analysis method with high sensitivity output such as CE-laser-induced florescence becomes an attractive alternative. Previously, a spacer-injection method with the use of zwitterionic surfactant (SB3-10) as a blocking agent to negate the cells’ mobility, induce aggregation and single microbial peak formation in a buffer solution was reported. Here, a fast, simple direct method for microbial detection in blood plasma without using the spacer and blocking agent is reported. To compensate for the natural electrophoretic heterogeneity of microbes, a CTAB additive was used to sweep all microbial cells towards the plasma peak where a single sharp microbial peak is formed and detected. With the use of BacLight™ Green bacterial stain, the microbial peak, generally, can be detected within 10 min in front of the plasma peak using capillary electrophoresis coupled with laser-induced florescence detection. The LOD of microbes detectable were 5 cells per injection. This technique provides a great advantage over traditional, time-consuming microbial inoculation methods.

Current perspectives on detection of microbial contamination in bioethanol fermentors

In recent years bioethanol has encompassed worldwide interest as a non-conventional bioenergy source. This fact has driven several bioethanol industries to produce more ethanol on a large scale via cost effective methods. However in the process of scaling up ethanol production bacterial contamination is becoming one of the more challenging problems facing the bioethanol industry. There are several traditional microbiological methods available to detect and subsequently limit these bacterial contaminants. These methods are time consuming, laborious and can be less sensitive. Consequently, it is necessary to find novel sensitive and economic detection methods to eradicate the contaminants long before they disrupt ethanol production. Molecular methods that can detect the contaminants even at very low numbers at any given stage would help in the design of more cost effective eradication strategies and better targeted antimicrobial treatments. Application of rapid molecular detection approaches have the potential to provide much more sensitive and rapid means to not only detect but quantitate microbial contaminants long before they become problematic to overall bioethanol formation.

Sterility testing by CE: A comparison of online preconcentration approaches in capillaries with greater internal diameters

Detection of microbial contamination is of critical importance in the medical and the food industry. Rapid tests for the absence or presence of viable microorganisms are in urgent demand. CE is a modern analytical technique that can be adapted for rapid screening of microbial contamination. However, the small dimensions of capillaries allow the introduction of only a small fraction of the sample, which can be problematic when examining large samples. In this article, we examine the possibilities of introducing larger sample volumes using capillaries with greater id together with different stacking techniques. The use of 0.32 mm id capillary and the injection of 60% of the capillary volume led to approximately 120-fold improvement of the injected sample volume over the classical injection 5% of a 0.10-mm id capillary. The setup we described opens new possibilities in sterility testing using CE.

Surface Charge Based Rapid Method for Detection of Microbial Contamination in Drinking Water and Food Products

Surface Charge Based Rapid Method for Detection of Microbial Contamination in Drinking Water and Food Products

Early detection of microbial contamination in processed tomatoes by electronic nose

Microbial contamination can easily affect processed tomato, thus determining both organoleptic adulterations and potential health risks for customers. Innovative techniques for a rapid and reliable diagnose of spoilage, such as electronic nose technology, are highly requested in order to guarantee food safety and to improve production. In this work canned peeled tomatoes were artificially spoiled with different kinds of microbial flora and then were analyzed by means of an electronic nose based on thin film metal oxide gas sensors. Preliminary analyses by dynamic-headspace gas chromatographic–mass spectrometry showed significant differences in the semi-quantitative volatile compounds profile of spoiled tomato samples just after few hours from contamination, thus suggesting to employ the electronic nose for an early diagnose of microbial presence. The electronic nose was indeed able to reveal contamination, even at early stages depending on the type of contaminant (e.g. for Saccharomyces cerevisiae and Escherichia coli), and to recognize spoiled tomato samples with good classification performances.

In situ detection and identification of microorganisms at single-colony resolution by spectral imaging

The in situ detection and identification of environmental microorganisms is important for general microbial ecology research. In addition, the rapid detection of microbial contamination in food-processing plants is essential. The current paper proposes a method for the rapid detection and identification of microorganisms using spectral imaging. Photosynthetic and non-photosynthetic bacterial colonies with different absorption spectra in the near-infrared wavelength region were measured directly from a Petri dish. The bacterial colonies were distinguished and subsequently typed using multiple discriminant analysis. The accuracy of the detection and identification of colonies of various sizes was evaluated. The results showed that colonies with diameters of 100 and 300 μm could be detected and identified, respectively, with adequate accuracy. Moreover, using our novel application of this spectral-imaging technique, the bacterial detection and identification times were reduced by more than half and by a few weeks, respectively, compared with conventional methods.

Artificial Odor Discrimination System Using Electronic Nose and Neural Networks for the Identification of Urinary Tract Infection

Current clinical diagnostics are based on biochemical, immunological, or microbiological methods. However, these methods are operator dependent, time-consuming, expensive, and require special skills, and are therefore, not suitable for point-of-care testing. Recent developments in gas-sensing technology and pattern recognition methods make electronic nose technology an interesting alternative for medical point-of-care devices. An electronic nose has been used to detect urinary tract infection from 45 suspected cases that were sent for analysis in a U.K. Public Health Registry. These samples were analyzed by incubation in a volatile generation test tube system for 4-5 h. Two issues are being addressed, including the implementation of an advanced neural network, based on a modified expectation maximization scheme that incorporates a dynamic structure methodology and the concept of a fusion of multiple classifiers dedicated to specific feature parameters. This study has shown the potential for early detection of microbial contaminants in urine samples using electronic nose technology.

RAPID DETECTION AND ENUMERATION OF CONTAMINANTS BY ATP BIOLUMINESCENCE USING THE MILLIFLEX® RAPID MICROBIOLOGY DETECTION AND ENUMERATION SYSTEM

ABSTRACT The Milliflex® Rapid Microbiology Detection and Enumeration System (RMDS) is an automated platform developed for the rapid detection of microbial contaminants in filterable samples. Based on membrane filtration, adenosine triphosphate bioluminescence and image analysis, RMDS gives microorganism enumeration a detection time reduced by a factor of four compared to plate count. After sample filtration and incubation step, a universal lysis solution and bioluminescence reagent are automatically sprayed across the membrane. The membrane is then analyzed in a detection tower where light emission is captured by a charged coupled device camera. Data collected are treated, directly providing a result in colony‐forming units. RMDS sensitivity is one cell and is equivalent to traditional microbiology. Rapid detection of bacteria, mixed bacteria and yeast was performed in various industrial samples. RMDS allowed rapid detection of contaminants in different simple and complex matrices such as water, tea, wine and mammalian cells. PRACTICAL APPLICATIONSThe Milliflex® Rapid Microbiology Detection and Enumeration System (RMDS) is used to detect and enumerate microorganisms in filterable samples with time to detection in hours.The easy handling of this detection platform enables its use in all kinds of industries from food and beverage industries to biotechnology or pharmaceutical industries where procedures and method validation are stricter. One application is the use of RMDS for purified water testing and rinsing water. Rapid enumeration of contaminants will improve the quality control analysis of industrial products. Rapid release of samples will result in the optimization of manufacturing and product testing, allowing high throughput and simultaneous analysis during the process.

The use of cationic surfactants and ionic liquids in the detection of microbial contamination by capillary electrophoresis

A rapid test of whether a laboratory sample contains any microorganisms is important and necessary for many areas of science and technology. Currently, most of the standard procedures for the detection of aerobic bacteria, anaerobic bacteria and fungi, require the preparation of microbial cultures in respective growth media, which are dramatically slow. Different approaches providing fast analysis such as CE are becoming more desired. To compensate for the natural electrophoretic heterogeneity of microbes, various buffer additives were examined to stack all bacteria and fungi in a sample plug into a single peak. This peak was removed from the molecular contaminants in the sample to prevent false positives. Both cationic surfactants and ionic liquids (IL) were investigated as run buffer additives and they are both widely applicable to different species of bacteria and fungi. Given that high concentrations of surfactants can potentially lyse cells, dicationic IL offer attractive auxiliary buffer additives for use in CE-based sterility tests. The analysis can be completed in 10 min, thus providing a great advantage over traditional direct inoculation methods that require several weeks to complete.