Chelate-Capped Nano-AgZn3 Dual Interphase Remodeling the Local Environment for Reversible Dendrite-Free Zinc Anode.

Rechargeable aqueous zinc-ion batteries (AZIBs) are among the most promising candidates for next-generation energy-storage devices. However, the large voltage polarisation and infamous dendrite growth hinder the practical application of AZIBs owing to their complex interfacial electrochemical environment. In this study, a hydrophobic zinc chelate-capped nano-silver (HZC-Ag) dual interphase is fabricated on the zinc anode surface using an emulsion-replacement strategy. The multifunctional HZC-Ag layer remodels the local electrochemical environment by facilitating the pre-enrichment and de-solvation of zinc ions and inducing homogeneous zinc nucleation, thus resulting in reversible dendrite-free zinc anodes. The zinc deposition mechanism on the HZC-Ag interphase is elucidated by density functional theory (DFT) calculations, dual-field simulations, and in situ synchrotron X-ray radiation imaging. The HZC-Ag@Zn anode exhibited superior dendrite-free zinc stripping/plating performance and an excellent lifespan of >2000 h with ultra-low polarisation of ≈17 mV at 0.5 mA cm-2 . Full cells coupled with a MnO2 cathode showed significant self-discharge inhibition, excellent rate performance, and improved cycling stability for >1000 cycles. Therefore, this multifunctional dual interphase may contribute to the design and development of dendrite-free anodes for high-performance aqueous metal-based batteries.

Comparison of Yersinia enterocolitica DNA Isolation from Swabs Without Pre-Enrichment on Selective Media With Isolation Preceded by Warm and Cold Enrichment.

Bacteriological methods for the identification of Yersinia enterocolitica are laborious and time-consuming. The aim of this study was to compare Y. enterocolitica DNA isolation from swabs without pre-enrichment on selective media with isolation preceded by warm and cold enrichment. The material for the study consisted of 150 rectal swabs taken from 50 clinically healthy fattening pigs. Forty-one Y. enterocolitica strains were isolated after warm enrichment and 43 after cold enrichment. The presence of ail, ystA, and yadA gene fragments was detected in all isolates. DNA isolation from swabs without pre-enrichment supported the detection of Y. enterocolitica in only nine samples. This study demonstrates that Y. enterocolitica DNA can be isolated from swabs without pre-enrichment on selective media, but this method is less sensitive than the approach where DNA isolation is preceded by warm or cold enrichment.

Selective pre-enrichment method to lessen time needed to recover Salmonella from commercial poultry processing samples.

Conventional Salmonella detection is time consuming, often employing a 24-h pre-enrichment step in buffered peptone water (BPW), followed by a 24-h selective enrichment in either Rappaport Vassiliadis (RV) or tetrathionate (TT) broths before streaking onto selective indicator agar. To reduce this time, we sought to optimize pre-enrichment for Salmonella recovery by evaluating the addition of selective chemicals to BPW. Duplicate samples each representative of 500 carcasses were collected by catching processing water drip under moving carcass shackle lines immediately after feather removal in each of nine commercial processing plants. Carcass drip samples were cultured under selective pre-enrichment conditions in parallel with BPW pre-enrichment followed by RV and TT selective enrichment. Addition of bile salts (1 g/L) and novobiocin (0.015 g/L) resulted in Salmonella recovery from 89% samples when plated directly after pre-enrichment compared to 67% recovery in non-selective BPW alone. Salmonella serovar identities were determined using CRISPR-SeroSeq. Overall, serovars matched between selective pre-enrichment and traditional enrichment methods. These data suggest that increasing the selectivity of Salmonella pre-enrichment step may lessen the need for a separate selective enrichment step thereby reducing time required for Salmonella isolation by 24 h.

Impact of desiccation and heat exposure stress on Salmonella tolerance to acidic conditions.

In a recent study, the pH of commonly used Salmonella pre-enrichment media became acidic (pH 4.0 to 5.0) when feed or feed ingredients were incubated for 24 h. Acidic conditions have been reported to injure or kill Salmonella. In this study, cultures of four known feed isolates (S. montevideo, S. senftenberg, S. tennessee, and S. schwarzengrund) and four important processing plant isolates (S. typhimurium, S. enteritidis, S. infantis, and S. heidelberg) were grown on meat and bone meal and later subjected to desiccation and heat exposure to stress the microorganism. The impact of stress on the isolates ability to survive in acidic conditions ranging from pH 4.0 to 7.0 was compared to the non-stressed isolate. Cell injury was determined on xylose lysine tergitol 4 (XLT4) and cell death determined on nutrient agar (NA). When measured by cell death in non-stressed Salmonella, S. typhimurium was the most acid tolerant and S. heidelberg was the most acid sensitive whereas in stressed Salmonella, S. senftenberg was the most acid tolerant and S. tennessee was the most acid sensitive. The pH required to cause cell injury varied among isolates. With some isolates, the pH required for 50% cell death and 50% cell injury was similar. In other isolates, cell injury occurred at a more neutral pH. These findings suggest that the pH of pre-enrichment media may influence the recovery and bias the serotype of Salmonella recovered from feed during pre-enrichment.

Evaluation and comparison of rapid methods for the detection of Salmonella in naturally contaminated pine nuts using different pre enrichment media.

Foodborne outbreaks, involving pine nuts and peanut butter, illustrate the need to rapidly detect Salmonella in low moisture foods. However, the current Bacteriological Analytical Manual (BAM) culture method for Salmonella, using lactose broth (LB) as a pre enrichment medium, has not reliably supported real-time quantitative PCR (qPCR) assays for certain foods. We evaluated two qPCR assays in LB and four other pre enrichment media: buffered peptone water (BPW), modified BPW (mBPW), Universal Pre enrichment broth (UPB), and BAX(®) MP media to detect Salmonella in naturally-contaminated pine nuts (2011 outbreak). A four-way comparison among culture method, Pathatrix(®) Auto, VIDAS(®) Easy SLM, and qPCR was conducted. Automated DNA extraction techniques were compared with manual extraction methods (boiling or InstaGene™). There were no significant differences (P > 0.05) among the five pre enrichment media for pine nuts using the culture method. While both qPCR assays produced significantly (P ≤ 0.05) higher false negatives in 24 h pre enriched LB than in the other four media, they were as sensitive as the culture method in BPW, mBPW, UPB, and BAX media. The VIDAS Easy and qPCR were equivalent; Pathatrix was the least effective method. The Automatic PrepSEQ™ DNA extraction, using 1000 µL of pre enrichment, was as effective as manual extraction methods.

Optimizing buffering chemistry to maintain near neutral pH of broiler feed during pre-enrichment for Salmonella.

Salmonella is a human pathogen that can accompany live broilers to the slaughter plant, contaminating fully processed carcasses. Feed is one potential source of Salmonella to growing broilers. Monitoring feed for the presence of Salmonella is part of good agricultural practice. The first step in culturing feed for Salmonella (which may be at low numbers and sub-lethally stressed) is to add it to a pre-enrichment broth which is incubated for 24 h. During the course of pre-enrichment, extraneous bacteria metabolize carbohydrates in some feed and excrete acidic byproducts, causing the pH to drop dramatically. An acidic pre-enrichment pH can injure or kill Salmonella resulting in a failure to detect, even if it is present and available to infect chickens. The objective of this study was to test an array of buffering chemistries to prevent formation of an injurious acidic environment during pre-enrichment of feed in peptone water. Five grams of feed were added to 45 mL of peptone water buffered with carbonate, Tris pH 8, and phosphate buffering ingredients individually and in combination. Feed was subjected to a pre-enrichment at 35°C for 24 h; pH was measured at 0, 18, and 24 h. Standard phosphate buffering ingredients at concentrations up to 4 times the normal formulation were unable to fully prevent acidic conditions. Likewise, carbonate and Tris pH 8 were not fully effective. The combination of phosphate, carbonate, and Tris pH 8 was the most effective buffer tested. It is recommended that a highly buffered pre-enrichment broth be used to examine feed for the presence of Salmonella.

High performance enrichment and analysis of fluoroquinolones residues in environmental water using cobalt ion mediated paper-based molecularly imprinted polymer chips.

BACKGROUND: Fluoroquinolones (FQs) are widely used for their excellent antimicrobial properties, yet their release into aquatic environments pose risks to ecosystems and public health. The accurate monitoring and analysis of FQs present challenges due to their low concentrations and the complex matrices found in actual environmental samples. To address the need for auto-pretreatment and on-line instrumental analysis, developing new microextraction materials and protocols is crucial. Such advancements will provide better analytical assurance for the effective extraction and determination of FQs at trace levels, which is of great significance to environmental protection and human health. RESULTS: In this work, we presented a Co2+ mediated paper-based molecularly imprinted polymer chip (CMC@Co-MIP), combined with UPLC analysis, to develop an effective analytical method for identifying and quantifying trace amounts of ciprofloxacin (CIP) and enrofloxacin (ENR) in water samples. Notably, the addition of Co2+ in CMC@Co-MIP helped to capture the template molecule CIP through coordination before imprinting, which significantly improved the ordering of the imprinted cavities. CMC@Co-MIP exhibited a maximum adsorption capacity up to 500.20 mg g-1 with an imprinting factor of 4.12, surpassing previous reports by a significant margin. Furthermore, the enrichment mechanism was extensively analyzed by various characterization techniques. The developed method showed excellent repeatability and reproducibility (RSD < 13.0 %) with detection limits ranging from 0.15 to 0.21 µg L-1 and recoveries ranging from 64.9 % to 102.3 % in real spiked water samples. SIGNIFICANCE: We developed a novel microextraction paper-based chip based on Co2+ mediation, which effectively improved the selectivity and convenience of extracting FQs. This breakthrough allowed the chip to have a high enrichment efficiency as well as provide a robust on-line instrumental program. It also confirms that the imprinting scheme based on metal ion coordination is a high-performance strategy.

A novel electrode for simultaneous detection of multiple heavy metal ions without pre-enrichment in food samples.

Integrating a pre-enrichment step into electrochemical detection methodologies has traditionally been employed to enhance the performance of heavy metal detection. However, this augmentation also introduces a degree of intricacy into the sensing process and increases energy consumption. In this work, Mo-doped WO3 is grown in situ on carbon cloth by one-step electrodeposition. The electrode detect multiple heavy metal ions simultaneously in the range of 0.1-100.0 µM with LODs ranging from 11.2 to 17.1 nM. The electrode successfully detected heavy metal ions in diverse food samples. This pioneering detection strategy realized the direct and simultaneous detection of multiple heavy metal ions by utilizing the valence property of WO3 and oxygen vacancies generated by molybdenum doping. The Mo-WO3/CC pre-enrichment-free detection electrode boasts straightforward preparation, a streamlined detection procedure, swift response kinetics, and superior performance relative to previously reported electrodes, which makes it possible to develop a portable heavy metal ion detection device.

Buffering Capacity Comparison of Tris Phosphate Carbonate and Buffered Peptone Water Salmonella Pre-Enrichments for Manufactured Feed and Feed Ingredients.

Various culture-based methods to detect Salmonella in animal feed have been developed due to the impact of this bacterium on public and animal health. For this project, tris phosphate carbonate (TPC) and buffered peptone water (BPW) buffering capacities were compared as pre-enrichment mediums for the detection of Salmonella in feed ingredients. A total of 269 samples were collected from 6 feed mills and mixed with the pre-enrichments; pH was measured before and after a 24 h incubation. Differences were observed when comparing pH values by sample type; DDGS and poultry by-product meal presented lower initial pH values for TPC and BPW compared to the other samples. For both TPC and BPW, meat and bone meal presented higher final pH values, while soybean meal and peanut meal had lower final pH values. Furthermore, for BPW, post cooling, pellet loadout, and wheat middlings reported lower final pH values. Additionally, most feed ingredients presented significant differences in pH change after 24 h of incubation, except DDGS. From meat and bone meal samples, four Salmonella isolates were recovered and identified: three using BPW and one using TPC. TPC provided greater buffer capacity towards neutral pH compared to BPW, but BPW was more effective at recovering Salmonella.

Pre-enriched saline gargle samples for detection of SARS-CoV-2.

A self­collected gargle sample, which avoids discomfort and largely reduces the dependency on medical resources, is emerging for detection of SARS­CoV­2. However, the incomplete usage of starting materials for both routine oropharyngeal swabs (OPS)/nasopharyngeal swabs (NPS) and saline gargle (SG) samples implies sensitivity can be further improved. Presented here is a bead­based strategy for pre­enrichment of SG samples, and results revealed that it acquired about 20 times the starting materials obtained from OPS samples for downstream detection of SARS­CoV­2. The sensitivity and specificity of this pre­enrichment strategy were validated in 100 paired pre­enriched saline gargle (PenSG) and OPS samples and 89 PenSG samples from healthy volunteers. In addition to detection of SARS­CoV­2, this pre­enrichment strategy may also be implemented in more clinical settings to optimize detection of other diseases.

Assessing Salmonella prevalence and complexity through processing using different culture methods.

Conventional Salmonella surveillance requires a week for isolation, confirmation, and subsequent serotyping. We previously showed that this could be reduced by 24 h by combining the pre-enrichment and enrichment steps into a single selective pre-enrichment step and was tested on directly after picking. The goal of this study was 2-fold: 1) to evaluate the use of selective pre-enrichment through each step of processing, including postintervention when the Salmonella load is reduced, and 2) to assess any changes in serovar populations in Salmonella positive samples. Duplicate carcass drip samples, each representative of 500 broiler carcasses, were collected by catching processing water drip under moving carcass shackle lines in each of three commercial broiler slaughter plants. Samples were collected post-pick, post-inside-outside bird wash (IOBW), and post-chill; duplicate wing rinses were performed pre- and post-antimicrobial parts dip. Each processing plant was sampled 6 times for a total of 180 samples collected. The number of Salmonella positives identified with selective pre-enrichment conditions (48/180) was similar to traditional selective enrichment culture conditions (52/180), showed good concordance in recovery rate between the 2 culture methods (Fisher's exact test, P = 0.72). We also found that the incidence of Salmonella reduced dramatically after antimicrobial intervention (post-pick 66.7% vs. post chill 8.3%). When serovar populations were evaluated in Salmonella positive samples using CRISPR-SeroSeq, we detected four different Salmonella serovars, Kentucky, Infantis, Schwarzengrund, and Typhimurium, and their incidence rose between post-pick and post-IOBW. The relative abundance of Infantis within individual samples increased between post-pick and post-IOBW while the relative abundance of the other 3 serovars decreased. These results suggest that a selective pre-enrichment step reduces the time required for Salmonella isolation without negatively affecting detection and serovar profiles in culture positive samples were not altered between culture conditions used.

FISH in Food Samples.

Foodborne diseases are a major global public health concern. The gold standard detection techniques, namely culture plating techniques, are nowadays considered inadequate for the modern food industry mainly due to the time requirements of this sector. As such, the adoption of faster detection methods to be routinely used in screening the protocols of foodborne pathogens is required. Fluorescence in situ Hybridization (FISH) methods have been described as a valid alternative to standard plating techniques and are compatible with the requirements of the food industry.Here, we give an overview of the methodological aspects to consider regarding sample preparation and sample analysis for pathogen detection in food matrices by FISH methodologies.

Metal-Organic Framework-Derived Hollow and Hierarchical Porous Multivariate Metal-Oxide Microspheres for Efficient Phosphoproteomics Analysis.

Pre-enrichment of the biological samples is a crucial step in phosphoproteomics research. At present, metal-oxide affinity chromatography (MOAC) is one of the most recognized enrichment strategy. Therefore, the design and preparation of a MOAC-based affinity material with better enrichment properties will be of great significance for the phosphoproteomics study. In this work, we obtained a novel multivariate metal-oxide microsphere (NiFe2O4@C@TiO2) with a hollow and hierarchical porous structure through pyrolysis of TiO2-modified Fe/Ni-based metal-organic frameworks (MOFs). After pyrolysis, the carbon matrix derived from the MOFs provided support and porous properties. Meanwhile, multivariate metal oxides endowed the microspheres with an excellent magnetic response property and superior enrichment performance for phosphorylated biomolecules. The unique hollow and hierarchical porous structure greatly enhanced the diffusion of phosphorylated biomolecules. Therefore, the microspheres exhibited excellent enrichment performance for phosphorylated biomolecules: a large adsorption capacity (124 µmol g-1), excellent selectivity (α-casein/BSA, 1:5000, m/m), perfect size-exclusion performance (α-casein digests/α-casein/BSA, 1:500:500), and extremely low detection limit (2 fmol). Furthermore, the microspheres showed excellent enrichment performance in a series of real biological samples, such as nonfat milk, serum, saliva, rat brain tissue, and plasma exosomes of patients with esophageal cancer, which further demonstrated its huge application potential in MS-based phosphoproteomics research.

An electrodialytic device for automated inorganic anion preconcentration with determination by ion chromatography-conductivity detection.

A 4-layer sandwiched device (4LSD) well suited for coupling to online ion chromatography (IC) systems was described and simultaneously performed target anion enrichment, matrix removal and sample injection within seconds. The basic assembly consisted of an extraction solution channel, a sample solution channel and two electrolyte channels. Cation-exchange resin (CER) was utilized to support the solution chamber, increase electrical conductivity and improve pressure resistance to achieve compatibility with a peristaltic pump. Filter placement ensured loop circulation of the 4LSD and prevented resin leakage. The 4LSD showed comparable performance to that of conventional solid-phase extraction (SPE) pretreatment in terms of matrix interference removal while enabling automation. The applied current, sample/extraction solution flow rate ratio, and initial concentration were discussed and optimized. Controllable 1-40-fold enrichment can be ensured. The migration phenomenon of different anions was discussed. F-, Cl-, NO2-, Br-, NO3-, SO42- and ClO4- exhibited satisfactory linear detection ranges within 2.5-1000 µg·L-1, and the calculated limits of detection (LODs) in milk formula were within the 0.097-0.79 mg·kg-1 range. The 4LSD was successfully applied to the determination of anions in milk formula with good spiked recoveries ranging between 92.54% and 107.2%, except for the NO2- recovery. The relative standard deviations (RSDs) ranged from 0.69% to 8.29%.

Development of a qPCR for the detection and quantification of Salmonella spp. in sheep feces and tissues.

Salmonella spp. are common causes of disease in intensive livestock production systems, and contamination of foodstuffs is of significant concern for public health. Therefore, the identification and quantification of Salmonella spp. is important for monitoring the level of fecal shedding or tissue colonization in infected animals and animal products. We developed and evaluated a quantitative PCR (qPCR) method on spiked sheep tissue and fecal samples for the detection and quantification of Salmonella spp. Without the use of a pre-enrichment step, the qPCR limit of detection (LOD) results for sheep fecal (4 × 104-6 × 103 cfu/g) and tissue (4 × 105-4 × 103 cfu/g) samples were not adequate for detection purposes. With the inclusion of a 6-h pre-enrichment step in buffered peptone water (BPW), the LOD was 9 cfu/g (2.57 × 101 copies/g) in sheep feces, and 5.4 cfu/g (3.22 copies/g) sheep tissue. Comparison of the 6-h BPW qPCR method with a 24-h mannitol-selenite-cystine broth enrichment culture method using spiked samples revealed a sensitivity of 91% and 92%, respectively, and a specificity of 100% for both methods. The correlation was significant between the quantity (copies/mL) of Salmonella spp. in BPW at 6 h and at 0 h, allowing semiquantitative analysis. Our results demonstrate that, following inclusion of a 6-h pre-enrichment step in BPW, qPCR is semiquantitative with improved LODs of Salmonella spp. in sheep fecal and tissue samples.

Detection of Salmonella Typhimurium and Listeria monocytogenes biofilm cells exposed to different drying and pre-enrichment times using conventional and rapid methods.

The capacity of real-time PCR (RT-PCR), the VIDAS immunoassay system, and the conventional count method for detecting Salmonella enterica serovar Typhimurium and Listeria monocytogenes biofilm cells was evaluated in this study. After biofilm formation, tests were performed under different drying times (0, 6, 12, 24, and 72 h) and pre-enrichment times (0, 6, 18, and 25 h). The direct epifluorescence microscopic results demonstrated that Salmonella Typhimurium and L. monocytogenes biofilm cells can remain viable for 72 h under drying conditions. Pre-enrichment time and type of medium played an essential role in the detection of both microorganisms after drying. Furthermore, RT-PCR was more sensitive than VIDAS and the conventional method for detecting Salmonella Typhimurium and L. monocytogenes cells at different drying times and without pre-enrichment (0 h), with a detection range between 102 and 107 CFU/mL. TSBYE-T80 used as a pre-enrichment medium was effective for detecting both bacteria and was more effective than Demi Fraser-T80 medium for detecting L. monocytogenes. Therefore, pre-enrichment is recommended to avoid false positives and false negatives due to the presence of dead cells or a very low initial concentration of cells after drying.

Detection of Listeria spp. and L. monocytogenes in pooled test portion samples of processed dairy products.

Listeria monocytogenes is a major foodborne pathogen. Testing multiple portions of the same final product is often required to verify the effectiveness of a food safety management system. Therefore, it will be advantageous to the laboratories to combine these test portions and process as one sample. However, combining samples for analysis, i.e., pooling, can be done only if there is no negative impact on the result. The objective of this study was to validate pooling of test portions for the detection of L. monocytogenes and Listeria spp. in dairy products as no scientific evidence currently exists to support this practice. Six representative matrices, namely, pudding, yogurt, brie cheese, 2% milk, ice cream and infant formula were spiked separately with stressed L. monocytogenes and Listeria spp. in 25 g and pooled test portions (375 g/250 g/125 g). Two methods, namely, ISO-11290-1:1996 Amd1:2004 and a validated alternative method Rapid'L.Mono were used for sample testing. Performance of a method in pooled test portions was considered to be satisfactory if the relative limit of detection (RLOD50; LOD50 [pooled test portion]/LOD50 [25 g test portion]) and limit of detection (LOD50) obtained was ≤2.5 and 1 CFU or MPN, respectively. Results obtained from L. monocytogenes and Listeria spp. trials were given equal weightage to decide on the impact of pooling. Acceptable RLOD50 and LOD50 values were consistently obtained in L. monocytogenes and Listeria spp. inoculation experiments when test portions were pooled up to 125 g for all matrices tested with both methods. While there was a slight delay for the primary enrichment of the pooled test portions to reach the desired incubation temperature when compared to the 25 g test portions, it did not negatively impact the outcome when samples were pooled up to 125 g. Background organisms were in general present at low concentrations and did not seem to adversely impact the recovery of the target organism in 125 g samples. Thus, pooling of test portions to up to 125 g for the detection of L. monocytogenes and Listeria spp. by two culture methods in processed dairy products has been validated.

Detection probability models for bacteria, and how to obtain them from heterogeneous spiking data. An application to Bacillus anthracis.

Efficient and correct evaluation of sampling results with respect to hypotheses about the concentration or distribution of bacteria generally requires knowledge about the performance of the detection method. To assess the sensitivity of the detection method an experiment is usually performed where the target matrix is spiked (i.e. artificially contaminated) with different concentrations of the bacteria, followed by analyses of the samples using the pre-enrichment method and the analytical detection method of interest. For safety reasons or because of economic or time limits it is not always possible to perform exactly such an experiment, with the desired number of samples. In this paper, we show how heterogeneous data from diverse sources may be combined within a single model to obtain not only estimates of detection probabilities, but also, crucially, uncertainty estimates. We indicate how such results can then be used to obtain optimal conclusions about presence of bacteria, and illustrate how strongly the sampling results speak in favour of or against contamination. In our example, we consider the case when B. cereus is used as surrogate for B. anthracis, for safety reasons. The statistical modelling of the detection probabilities and of the growth characteristics of the bacteria types is based on data from four experiments where different matrices of food were spiked with B. anthracis or B. cereus and analysed using plate counts and qPCR. We show how flexible and complex Bayesian models, together with inference tools such as OpenBUGS, can be used to merge information about detection probability curves. Two different modelling approaches, differing in whether the pre-enrichment step and the PCR detection step are modelled separately or together, are applied. The relative importance on the detection curves for various existing data sets are evaluated and illustrated.

Validation of test portion pooling for Salmonella spp. detection in foods.

Pathogen monitoring programs play a crucial role in the verification of the effectiveness of implemented hygiene control measures. Sampling and testing procedures included in pathogen monitoring involve the analysis of multiple test portions where all samples must be negative for the presence of pathogens for a certain test portion size. Many food safety programs require increased testing due to the risks that a pathogen may be present. Analyzing more than one test portion could prove to be expensive and labor intensive. When more than one test portion for a specified food item is to be tested, the test portions could be combined to form a pooled test portion to reduce laboratory workload, costs of reagents and further confirmatory steps, but only when evidence is available that pooling does not affect on the number of false negative results for different matrices. This study has been performed to demonstrate the equivalence of test portion pooling for Salmonella detection with five different methods using cultural, ELISA and Real Time PCR technologies. Twenty-three (23) different food items including confectionary products, meal components, infant formula, pet food and powdered beverages were validated. Other complementary parameters like impact of minimum and maximum incubation time for pre-enrichment, temperature profile, pH and Salmonella concentration after the pre-enrichment and background flora have also been considered in the study. The results showed that pooling test portions up to 375g for Salmonella detection is valid for the methods that were tested. Relative level of detection (RLOD50) values for 22 of the food items tested were acceptable (i.e. lower than 2.5) when comparing the reference sample size (25g) against the alternative pooled sample size (375g), provided the enrichment broth was pre-warmed and maximum incubation time is respected.

Quantification of Growth of Campylobacter and Extended Spectrum ß-Lactamase Producing Bacteria Sheds Light on Black Box of Enrichment Procedures.

Campylobacter is well recognized as the leading cause of bacterial foodborne diarrheal disease worldwide, and is routinely found in meat originating from poultry, sheep, pigs, and cattle. Effective monitoring of Campylobacter contamination is dependent on the availability of reliable detection methods. The method of the International Organization for Standardization for the detection of Campylobacter spp. in food (ISO 10272-1:2006) recommends the use of Bolton broth (BB) as selective enrichment medium, including a pre-enrichment step of 4-6 h at 37°C to revive sublethally damaged cells prior to incubation for 2 days at 41.5°C. Recently the presence of abundantly growing extended spectrum ß-lactamase producing Enterobacteriaceae (ESBL bacteria) has become one of the most important factors that interfere with the isolation of Campylobacter, resulting in false-negative detection. However, detailed growth dynamics of Campylobacter and its competitors remain unclear, where these would provide a solid base for further improvement of the enrichment procedure for Campylobacter. Other enrichment broths, such as Preston broth (PB) and BB plus clavulanic acid (BBc) have been suggested to inhibit competitive flora. Therefore, these different broths were used as enrichments to measure the growth kinetics of several strains of Campylobacter jejuni and ESBL bacteria separately, in co-culture and of strains in chicken samples. The maximum cell numbers and often the growth rates of Campylobacter in mixed culture with ESBL bacteria were significantly lower than in single cultures, indicating severe suppression of Campylobacter by ESBL bacteria, also in naturally contaminated samples. PB and BBc successfully diminished ESBL bacteria and might therefore be a better choice as enrichment medium in possibly ESBL-bacteria contaminated samples. The efficacy of a pre-enrichment step in the BB ISO-procedure was not supported for cold-stressed and non-stressed cells. Therefore, omission of this step (4-6 h at 37°C) might be advised to obtain a less troublesome protocol.