Enteric bacteriophages as potential fecal indicators in ground beef and poultry meat.

Recovery efficiencies of enteric bacteriophages (F+ RNA coliphages, somatic coliphages, and Salmonella phages) as alternative fecal indicators were determined from ground beef and chicken breast meat using amino acid eluants (glycine and threonine) and a complex eluant (3% beef extract). Levels of F+ RNA coliphages (MS2, GA, Qbeta, FI, and SP), the somatic coliphage phiX174, and three environmental isolates of Salmonella phages (isolated from raw sewage) were assayed using three respective hosts: Escherichia coli Famp, E. coli C, and Salmonella Typhimurium. When 8% polyethylene glycol and 0.1 M NaCl were used to precipitate bacteriophages eluted with five different eluants, the highest recoveries of the three phage groups were with 0.5 M threonine and 0.25 M glycine-threonine. The average recoveries of F+ RNA coliphages, somatic coliphages, and the Salmonella phages from ground beef and chicken meat were 100, 69, and 65%, respectively, with threonine (0.5 M, pH 9.0) as the eluate. Of eight market food samples tested, F+ RNA coliphages were detected in five (63%) and somatic coliphages were detected in seven (88%). The overall detection sensitivity of the method was 3 PFU/100 g of ground beef or chicken meat. Levels of bacteriophages and bacterial indicators on chicken carcass surfaces were determined at identified critical control points at a poultry plant. Through the processing steps of evisceration, washing, and chilling, the levels of F+ RNA coliphages and fecal coliforms were reduced by 1.6 and 1.9 log10 PFU or CFU/100 g, respectively. F+ RNA coliphages and perhaps other enteric bacteriophages may be effective candidate indicators for monitoring the microbiological quality of meat, poultry, and perhaps other foods during processing. The bacteriophage concentration method developed provides a simple, rapid, and practical tool for the evaluation of fecal contamination levels in ground beef and processed chicken meat.     

Particle-size distribution as indicator for fecal bacteria contamination of drinking water from karst springs

Continuous monitoring of particle-size distribution (PSD), total organic carbon (TOC), turbidity, discharge and physicochemical parameters, together with analyses of fecal indicator bacteria, particularly Escherichia coli, made it possible to better understand the processes governing pathogen transport in karst groundwater and to establish PSD as indicator for possible microbial contamination of drinking water from karst springs. In the study area near Yverdon-les-Bains, Switzerland, tracer tests proved connection between a sinking stream draining agricultural land and several springs, 4.8-6.3 km away. Tracing and monitoring results demonstrate that (i) suspended particles (turbidity) in the spring water either originate from remobilization of sediments inside the aquifer (autochthonous) or from the sinking stream and land surface (allochthonous); (ii) allochthonous turbidity coincides with increased E. coli and TOC levels; (iii) PSD makes it possible to distinguish the two types of turbidity; (iv) a relative increase of finer particles (0.9-10 microm) indicates allochthonous turbidity and thus possible fecal contamination. The method permits to optimize water treatment and identify periods when the spring water must be rejected. Findings from other test sites confirm the feasibility of this approach.     

Raman microspectroscopy-based identification of individual fungal spores as potential indicators of indoor contamination and moisture-related building damage

We present an application of Raman microspectroscopy (RMS) for the rapid characterization and identification of individual spores from several species of microfungi. The RMS-based methodology requires minimal sample preparation and small sample volumes for analyses. Hence, it is suitable for preserving sample integrity while providing micrometer-scale spatial resolution required for the characterization of individual cells. We present the acquisition of unique Raman spectral signatures from intact fungal spores dispersed on commercially available aluminum foil substrate. The RMS-based method has been used to compile a reference library of Raman spectra from several species of microfungi typically associated with damp indoor environments. The acquired reference spectral library has subsequently been used to identify individual microfungal spores through direct comparison of the spore Raman spectra with the reference spectral signatures in the library. Moreover, the distinct peak structures of Raman spectra provide detailed insight into the overall chemical composition of spores. We anticipate potential application of this methodology in the fields of public health, forensic sciences, and environmental microbiology.     

Nicotine derivatives in wastewater and surface waters: application as chemical markers for domestic wastewater

Nicotine is extensively metabolized in the human body to a number of compounds, which may enter natural waters via discharge of domestic wastewater. However, little is known on exposure of and potential effects on the aquatic environment. In this study, two major urinary metabolites, cotinine and 3'-hydroxycotinine, as well as a further tobacco alkaloid, N-formylnornicotine, were measured in wastewater and water from Swiss lakes using an analytical procedure based on SPE and LC-MS/MS SRM with cotinine-d3 as internal standard (LOQs, 1.0-1.5 ng/L). Typical concentrations of cotinine and 3'-hydroxycotinine were approximately 1-10 microg/L in untreated wastewater, but clearly less in treated wastewater (approximately 0.01-0.6 microg/L), corresponding to elimination efficiencies of 90-99%. N-Formylnornicotine, however,was found at similar concentrations in untreated and treated wastewater (0.02-0.15 microg/L). Its apparent persistence during wastewater treatment was further confirmed by incubation experiments with activated sludge. In lakes, cotinine, 3'-hydroxycotinine, and N-formylnornicotine were detected at concentrations up to 15, 80, and 6 ng/L, respectively. Concentrations in lakes correlated with the expected anthropogenic burden by domestic wastewater (ratio population per water throughflow), demonstrating the suitability of these nicotine derivatives as hydrophilic, anthropogenic markers. In small receiving waters with significant wastewater discharges, concentrations of a few hundred ng/L may be expected. Possible ecotoxicological risks associated with such environmental concentrations, can, however, not be assessed at present as data on effects on aquatic organisms are very limited, in particular on long-term effects.     

Metabolomic profiling of food matrices: Preliminary identification of potential markers of microbial contamination

The research aimed to generate an early warning system highlighting in real-time bacterial contamination of meat matrices and providing information which could support companies in accepting or rejecting batches. Current microorganisms’ detection methods rely on techniques (plate counting), which provide retrospective values for microbial contamination. The purpose of this research was to evaluate the ability of the headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC/MS) methodologies to detect volatile organic carbons (VOCs), which may be associated to a peculiar microbiological contamination of food. The disposal of fast headspace gas chromatography-mass spectrometry (HS-SPME-GC/MS) able to accurately and rapidly (30 min per sample) detect pathogens in raw meat could replace the traditional and time-consuming (3 to 4 days) standardized microbiological analysis required by regulations. Experiments focused on qualitative and quantitative evaluations of VOCs produced by Salmonella Typhimurium, Campylobacter jejuni, and Staphylococcus aureus in different types of raw meat (beef, pork, chicken). HS-SPME-GC/MS allowed to use smaller sample volumes compared to traditional methods with no sample processing and the potentiality for its application on various food matrices for the detection of a wide variety of pathogens. Data analysis showed the identification of unique VOCs’ profiles being possible markers of meat contamination due to their association to specific pathogens. The identification of VOCs markers in association to selected bacterial pathogens and their metabolites could support the rapid determination of specific meat samples contamination. Further research is required to outline-specific metabolic profiles for each microorganism responsible of meat contamination and prevent false positives.     

New phytochemicals as potential human anti-aging compounds: Reality,promise, and challenges

Aging is an inevitable process influenced by genetic, lifestyle, and environmental factors. Indirect evidence shows that several phytochemicals can have anti-aging capabilities, although direct evidence in this field is still limited. This report aims to provide a critical review on aspects related to the use of novel phytochemicals as anti-aging agents, to discuss the obstacles found when performing most anti-aging study protocols in humans, and to analyze future perspectives. In addition to the extensively studied resveratrol, epicatechin, quercetin, and curcumin, new phytochemicals have been reported to act as anti-aging agents, such as the amino acid L-theanine isolated from green tea, and the lignans arctigenin and matairesinol isolated from Arctium lappa seeds. Furthermore, this review discusses the application of several new extracts rich in phytochemicals with potential use in anti-aging therapies. Finally, this review also discusses the most important biomarkers to test anti-aging interventions, the necessity of conducting epidemiological studies and the need of clinical trials with adequate study protocols for humans.     

Lactobacilli isolated from chicken intestines: potential use as probiotics

Lactobacillus strains were tested for their in vitro probiotic properties. Cell surface hydrophobicity was found to be very high for Lactobacillus fermentum subsp. cellobiosus and Salmonella Gallinarum; high values could indicate a greater ability to adhere to epithelial cells. Studies on Lactobacillus animalis indicated relative cell surface hydrophobicities smaller than those of L. fermentum subsp. cellobiosus and L. fermentum. L. animalis and Enterococcus faecalis were able to coaggregate with L. fermentum subsp. cellobiosus and L. fermentum, respectively, but not with Salmonella Gallinarum. After mixed-culture studies for determining suitable growth behavior, the pair of strains L. animalis plus L. fermentum subsp. cellobiosus was selected for an attempted challenge against Salmonella Gallinarum. Double and triple mixed-culture studies indicated that selected lactobacillus strains were able to retain their beneficial characteristics in the presence of Salmonella Gallinarum such as presence of lectins, production of antimicrobial compounds, and ability to grow and compete. The selected microorganisms can be considered as potential ingredients for a chicken probiotic feed formulation intended to control salmonellosis and also improve poultry sanitation.     

Use of phenolic compounds from olive mill wastewater as valuable ingredients for functional foods

ABSTRACT

Olive mill wastewater (OMW) is a pollutant by-product from the virgin olive oil production. Its high content in phenolic compounds makes them play an important role for their use in foods, for their high antioxidant significance. The present paper gives an overview on the techniques for OMW valuable ingredient separation, focusing on the most effective ones for their use in food products as functional ingredients. We report on effective methods to recover OMW phenolics, and give several examples on the use these extracts in foods. When added into vegetable oils, their effect on retarding lipid oxidation improves the oxidative status of the product, whilst several challenges need to be faced. OMW phenolic extracts were also used in food emulsions, milk products or other model systems, showing promising results and little or no negative impact on the sensory characteristics or other properties. Their possible use as antimicrobial agents is also another promising approach, as positive results were obtained when applied in meat products. Other examples of using natural phenolic extracts from other sources are suggested also for OMW extracts, to expand their use and thus to improve the nutritional and technological quality of foods.     

Olive mill wastewater as a potential source of antibacterial and anti-inflammatory compounds against the food-borne pathogen Campylobacter

Phenolics and secoiridoids from crude olive mill wastewater (OMW) were purified using a preparative hybrid separation process: pressure-driven cross-flow membrane micro-(MF) and ultrafiltration (UF) and solid phase extraction (SPE). Five fractions with different contents of these compounds were obtained and their antibacterial activity against several strains of Campylobacter was evaluated. The results showed that the fraction with the highest content of phenolic and secoiridoid compounds had a relevant antibacterial activity against all tested strains with a strain-dependent character. Pre-treatment of LPS-stimulated cells with this fraction decreased the production of nitric oxide (NO), showing its potential as anti-inflammatory agent. Finally, the effect of the same fraction on the viability of human intestinal epithelial cell lines was evaluated. In conclusion, OMW could be useful as a potential source of bioactive compounds against Campylobacter, increasing the valorisation of olive oil by-products.     

食用槟榔加工工艺及其化学与微生物污染研究进展

论述目前食用槟榔加工主要采取的工艺方法、发展趋势及其存在的化学及微生物污染风险,阐述目前槟榔加工过程中质量安全控制技术研究现状,指出未来食用槟榔加工技术的研究重点。     

Optimal fluorescence excitation and emission bands for detection of fecal contamination

Fecal contamination of food products is a critical health issue. To test the feasibility of the use fluorescent techniques to detect fecal contamination, fluorescence excitation and emission characteristics of fecal matter from cows, deer, swine, chickens, and turkeys in the UV to far-red regions of the spectrum were evaluated. To allow the optimization of the detection of fecal contamination on animal carcasses and cut meats, emission-excitation spectra of the feces were compared with spectra for animal meats. The feedstuffs for the swine, chickens, and turkeys were also analyzed. Excitation at approximately 410 to 420 nm yielded the highest level of fluorescence for both feces and feedstuffs. Emission maxima were in the red region (at 632 nm for chicken feces and at 675 nm for the feces of the other species). The major constituent responsible for emission at 632 nm was tentatively identified as protoporphyrin IX; emission at 675 nm most likely emanates from chlorophyll a or its metabolites. Animal meats emitted strong fluorescence in the blue-green regions, but no emission peaks were observed in the red region for these meats. These results suggest that fluorescence emissions from naturally occurring chlorophyll a and its metabolites are good markers for fecal contamination and that with excitation at 410 to 420 nm, the responses of fecal matter can easily be differentiated from the responses of animal meats. We suggest that the detection of fecal contamination can be enhanced by requiring a minimum chlorophyll a content in the finishing diets of all farm animals.     

Effects of pharmaceutically active compounds on a mixed microbial community originating from a municipal wastewater treatment plant

The growth and composition of microorganisms found in a municipal wastewater treatment plant were investigated in the presence of four pharmaceutically active compounds (PhACs) [ketoprofen, naproxen, carbamazepine, and clofibric acid] in batch reactors at varying organic loadings. Overall, the data suggest that microbial growth inhibition was correlated to organic loading rather than PhAC concentration. Significant inhibition ranging from 34 to 43% was observed under the lowest organic loading for all PhACs other than clofibric acid. No inhibition was observed at the highest organic loading. Higher microbial inhibition was not observed with increased PhAC concentration for a given organic loading. These results indicatethatthe presence of PhAC may affect microbial growth especially under lower organic loading conditions. Further validation is required with additional PhACs, organic substrates, and a wider loading range. In addition, significant microbial shifts were observed in the presence of ketoprofen and naproxen. These data suggest that, in addition to their effect on overall microbial growth, PhACs may affect the microbial ecology and additional research should be carried out to identify PhACs that have the potential of affecting ecologically important microorganisms in wastewater treatment processes and aquatic environments in general.     

Phylogenetic and functional biomakers as indicators of bacterial community responses to mixed-waste contamination

Few studies have demonstrated changes in community structure along a contaminant plume in terms of phylogenetic, functional, and geochemical changes, and such studies are essential to understand how a microbial ecosystem responds to perturbations. Clonal libraries of multiple genes (SSU rDNA, nirK, nirS, amoA, pmoA, and dsrAB) were analyzed from groundwater samples (n = 6) that varied in contaminant levels, and 107 geochemical parameters were measured. Principal components analyses (PCA) were used to compare the relationships among the sites with respect to the biomarker (n = 785 for all sequences) distributions and the geochemical variables. A major portion of the geochemical variance measured among the samples could be accounted for by tetrachloroethene, 99Tc, No3, SO4, Al, and Th. The PCA based on the distribution of unique biomarkers resulted in different groupings compared to the geochemical analysis, but when the SSU rRNA gene libraries were directly compared (deltaC(xy) values) the sites were clustered in a similar fashion compared to geochemical measures. The PCA based upon functional gene distributions each predicted different relationships among the sites, and comparisons of Euclidean distances based upon diversity indices for all functional genes (n = 432) grouped the sites by extreme or intermediate contaminant levels. The data suggested that the sites with low and high perturbations were functionally more similar than sites with intermediate conditions, and perhaps captured the overall community structure better than a single phylogenetic biomarker. Moreover, even though the background site was phylogenetically and geochemically distinct from the acidic sites, the extreme conditions of the acidic samples might be more analogous to the limiting nutrient conditions of the background site. An understanding of microbial community-level responses within an ecological framework would provide better insight for restoration strategies at contaminated field sites.     

Escalating water demand for energy production and the potential for use of treated municipal wastewater

To ensure sufficient thermoelectric power production in the future, the use of alternative water sources to replace freshwater consumption in power plants will be required. The amount of municipal wastewater (MWW) being produced and its widespread availability merit the investigation of this potential source of cooling water. This is particularly important for thermoelectric power plants in regions where freshwater is not readily available. Critical regulatory and technical challenges for using MWW as makeup water in recirculating cooling systems are examined. The existing regulations do not prohibit wastewater reuse for power plant cooling. The challenges of controlling corrosion, mineral scaling, and biofouling in recirculating cooling systems need to be carefully considered and balanced in a holistic fashion. Initial investigations suggest that many of these challenges can be surmounted to ensure the use of MWW in recirculating cooling systems.     

Characterization of homofermentative lactobacilli isolated from kefir grains: potential use as probiotic

Considering that several health promoting properties are associated with kefir consumption and a reliable probiotic product requires a complete identification of the bacterial species, the present work evaluates several proved markers of probiotic potential of eleven isolates of homofermentative lactobacilli isolated from kefir grains and molecular identification and genotypic diversity. Using restriction analysis of amplified ribosomal DNA (ARDRA) and analysis of the 16S-23S rRNA internal spacer region we confirmed that all homofermentative lactobacilli belong to the species Lactobacillus plantarum. RAPD-PCR analysis allowed the discrimination of lactobacilli in five clusters. All isolates exhibited high resistance to bile salt. High survival after one hour of exposure to pH 2.5 was observed in Lb. plantarum CIDCA 8313, 83210, 8327 and 8338. All isolates were hydrophilic and non autoaggregative. Isolate CIDCA 8337 showed the highest percentage of adhesion among strains. All tested lactobacilli had strong inhibitory power against Salmonella typhimurium and Escherichia coli. Seven out of eleven isolates showed inhibition against Sal. enterica and five isolates were effective against Sal. gallinarum. Only CIDCA 8323 and CIDCA 8327 were able to inhibit Sal. sonnei. We did not find any correlation between the five clusters based on RAPD-PCR and the probiotic properties, suggesting that these isolates have unique characteristics.     

Selection of Lactobacillus strains from fermented sausages for their potential use as probiotics

A rapid screening method was used to isolate potentially probiotic Lactobacillus strains from fermented sausages after enrichment in MRS broth at pH 2.5 followed by bile salt stressing (1% bile salts w/v). One hundred and fifty acid- and bile-resistant strains were selected, avoiding preliminary and time-consuming isolation steps. Strains were further characterized for survival at pH 2.5 for 3 h in phosphate-buffered saline and for growth in the presence of 0.3% bile salts with and without pre-exposure at low pH. Twenty-eight strains showed a survival >80% at pH 2.5 for 3 h; moreover, most of the strains were able to grow in the presence of 0.3% bile salts. Low pH and bile resistance was shown to be dependent on both the species, identified by phenotypic and molecular methods, and the strain tested. This is the first report on the direct selection of potentially probiotic lactobacilli from dry fermented sausages. Technologically interesting strains may be used in the future as probiotic starter cultures for novel fermented sausage manufacture.     

Xylan as by-product of biorefineries: Characteristics and potential use for food applications

The conversion or biorefinery of renewable resources gives three main components: cellulose, lignin and hemicelluloses. Xylans are part of the hemicelluloses fraction and of a certain use in nutrition, food technology, new materials and biotechnology. The functionality is based on certain xylan characteristics giving the polysaccharide unique hydrophilic as well as hydrophobic interaction capacities. The rheology of xylan systems indicates that the properties for some conformations of this xylose based macromolecule depend also on associated water clusters or the integration of water molecules. Several applications for the use of the functionality of the hydrocolloid are presented.