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The food industry is unceasingly searching for accurate ways to check food safety. The presence of microbial toxins such as bacterial toxins, fungal toxins, and algal toxins in food products has become a concerning issue worldwide. These toxins can contaminate food during various stages of its production, transportation, and storage, posing a serious threat to human health. The reliable and quick determination of microbial toxins has become significant for eliminating the disease outbreaks caused by contaminated foodstuffs. Thus, there is an increased interest in developing low-cost and high-throughput sensors for the determination of microbial toxins in food matrices. Recently, fluorescent sensors have gained importance as one of the effective alternatives in food safety monitoring. The use of nanomaterials in the fabrication of fluorescent sensors has imparted amazing properties such as high selectivity and sensitivity, rapid results, good stability, and reproducibility to analyze microbial toxins in real food matrices. The present review summarizes the recent developments in nanomaterial-based fluorescent sensors for the detection of microbial toxins in food products. The review delves into the details of various fluorescent nanomaterials such as metal-based nanomaterials, upconversion nanoparticles, semiconductor quantum dots, carbon-based nanomaterials, and luminescent metal-organic frameworks describing their unique properties, advantages, drawbacks, and applications in the detection of microbial toxins in food products. Also, the performance comparison of the fluorescent nanomaterials in the detection of these toxins has been discussed. Further, the commercial aspects of fluorescent nanosensors for the detection of microbial toxins have been included, along with the future insights and challenges associated with microbial toxin detection. [ABSTRACT FROM AUTHOR]

Abstract Secretion systems play a crucial role in microbe-microbe or host-microbe interactions. Among these systems, the extracellular contractile injection system (eCIS) is a unique bacterial and archaeal extracellular secretion system that injects protein toxins into target organisms. However, the specific proteins that eCISs inject into target cells and their functions remain largely unknown. Here, we developed a machine learning classifier to identify eCIS-associated toxins (EATs). The classifier combines genetic and biochemical features to identify EATs. We also developed a score for the eCIS N-terminal signal peptide to predict EAT loading. Using the classifier we classified 2,194 genes from 950 genomes as putative EATs. We validated four new EATs, EAT14-17, showing toxicity in bacterial and eukaryotic cells, and identified residues of their respective active sites that are critical for toxicity. Finally, we show that EAT14 inhibits mitogenic signaling in human cells. Our study provides insights into the diversity and functions of EATs and demonstrates machine learning capability of identifying novel toxins. The toxins can be employed in various applications dependently or independently of eCIS.

Insect host defense comprises two complementary dimensions, microbial killing-mediated resistance and microbial toxin neutralization-mediated resilience, both jointly providing protection against pathogen infections. Insect defensins are a class of effectors of innate immunity primarily responsible for resistance to Gram-positive bacteria. Here, we report a newly originated gene from an ancestral defensin via genetic deletion following gene duplication in Drosophila virilis, which confers an enhanced resilience to Gram-positive bacterial infection. This gene encodes an 18-mer arginine-rich peptide (termed DvirARP) with differences from its parent gene in its pattern of expression, structure and function. DvirARP specifically expresses in D. virilis female adults with a constitutive manner. It adopts a novel fold with a 310 helix and a two CXC motif-containing loop stabilized by two disulfide bridges. DvirARP exhibits no activity on the majority of microorganisms tested and only a weak activity against two Gram-positive bacteria. DvirARP knockout flies are viable and have no obvious defect in reproductivity but they are more susceptible to the DvirARP-resistant Staphylococcus aureus infection than the wild type files, which can be attributable to its ability in neutralization of the S. aureus secreted toxins. Phylogenetic distribution analysis reveals that DvirARP is restrictedly present in the Drosophila subgenus, but independent deletion variations also occur in defensins from the Sophophora subgenus, in support of the evolvability of this class of immune effectors. Our work illustrates for the first time how a duplicate resistance-mediated gene evolves an ability to increase the resilience of a subset of Drosophila species against bacterial infection. [ABSTRACT FROM AUTHOR]

Frequent contamination of foods with microbial toxins produced by microorganisms such as bacteria, fungi, and algae represents an increasing public health problem that requires the development of quick and easy tools to detect them at trace levels. Recently, it has been found that colorimetric detection methods may replace traditional methods in the field because of their ease of use, quick response, ease of manufacture, low cost, and naked-eye visibility. Therefore, it is suitable for fieldwork, especially for work in remote areas of the world. However, the development of colorimetric detection methods with low detection limits is a challenge that limits their wide applicability in the detection of food contaminants. To address these challenges, nanomaterial-based transduction systems are used to construct colorimetric biosensors. For example, gold nanoparticles (AuNPs) provide an excellent platform for the development of colorimetric biosensors because they offer the advantages of easy synthesis, biocompatibility, advanced surface functionality, and adjustable physicochemical properties. The selectivity of the colorimetric biosensor can be achieved by the combination of aptamers and gold nanoparticles, which provides an unprecedented opportunity to detect microbial toxins. Compared to antibodies, aptamers have significant advantages in the analysis of microbial toxins due to their smaller size, higher binding affinity, reproducible chemical synthesis and modification, stability, and specificity. Two colorimetric mechanisms for the detection of microbial toxins based on AuNPs have been described. First, sensors that use the localized surface plasmon resonance (LSPR) phenomenon of gold nanoparticles can exhibit very strong colors in the visible range because of changes caused by aggregation or disaggregation. Second, the detection mechanism of AuNPs is based on their enzyme mimetic properties and it is possible to construct a colorimetric biosensor based on the 3,3',5,5'-tetramethylbenzidine/Hydrogen peroxide, TMB/H2O2 reaction to detect microbial toxins. Therefore, this review summarizes the recent applications of AuNP-based colorimetric aptasensors for detecting microbial toxins, including bacterial toxins, fungal toxins, and algal toxins focusing on selectivity, sensitivity, and practicality. Finally, the most important current challenges in this field and future research opportunities are discussed. [ABSTRACT FROM AUTHOR]

Currently, investigations on the microbiota of sports centers and related facilities have been carried out in some countries, which showed that Microsporum gypseum, Trichophyton mentagrophytes and T. tonsurans are important dermatoprotofungi. In China, some research on athletes and sports equipment between the fungal community and public health has made some interesting achievements. However, the bacterial group among them has not been reported. Therefore, The aim of this study was to uncover (I) gymnastic equipment is there potential pathogenic factors and (ii) is there any difference in the biomarker of bacterial in different types of gymnastic room? The samples were collected from the gymnastics halls of one university in western China and main sports equipment, including gymnastics carpets, moving barres, hoops and balls, as well as wall bars, parallel bars and horizontal bars. The 16S rDNA of all the samples was sequenced, and the analyses were performed using FaproTax, Bug base function prediction and Line Discriminant Analysis (LDA) Effect Size. A 16S rDNA sequence analysis revealed abundant bacterial species biodiversity on gymnasts and apparatuses from two gymnastics halls at a university in western China. An analysis using the FaproTax and Bugbase functional prediction platforms showed that there were some opportunistic pathogens on the athletes and equipment from the Rhythmic Gymnastics (RG) and Artistic Gymnastics (AG) halls, such as Staphylococcus and Corynebacteiaceae. Infectious agents associated with cancer induction and development, such as Ruminococcaceae, Veillonellaceae and Moraxellaceae, as well as microbial toxin producers with a potential impact on human health, were also detected. According to a line discriminant analysis (LDA effect size), the bacterial biomarker groups of the two gymnasiums were different at the phylum-genus level: for RG, Erysipelatoclostridium, Lachnospiraceae and Bacteroidales, while for AG, Rhizobiales. Based on the results of the investigation, we suggest that more comprehensive consideration should be given to indoor microbial biodiversity and related public health problems in school gymnasiums. [ABSTRACT FROM AUTHOR]

Parkinson's disease (PD) is a chronic, progressive and second most prevalent neurological disorder affecting the motor system. Cardinal motor impairment and α-synucleinopathy are the characteristic features of PD. Recently, it has been identified that the gut-brain axis is substantially regulated by the gut microbiome (GM) through an immunological, neuroendocrine, and neural mechanism. However, disturbance in the gut-microbiome-brain axis in PD might proceed to gastrointestinal manifestations intermittently leading to the motor system and the PD pathogenesis itself. The gut microbial toxins may induce the production of α-synuclein (α-syn) aggregates in the enteric nervous system (ENS), which may proliferate and propagate in a prion-like-manner through the vagus nerve to the central nervous system (CNS); supporting the hypothesis that, GM might play a pivotal role in PD pathogenesis. Overstimulated innate immune system due to intestinal bacterial overgrowth or gut dysbiosis and the enhanced intestinal permeability may persuade systemic inflammation, while the activation of enteric glial cells and enteric neurons may contribute to α-synucleinopathy. Gut microbiota can bear a significant impact on neurological outcomes such as learning, memory and cognition. In this review paper, we summarize how the alterations in gut microbiota and ENS inflammation are associated with PD pathogenesis. The evidence supporting the causative role played by gut-associated dysbiosis and microbial byproducts, in the onset of PD is also discussed. We have highlighted the landmark discoveries in the field of PD particularly focusing on the gut-brain axis. A better comprehension of the interaction between the gut-brain axis, gut microbiota, and PD can usher in novel therapeutic and diagnostic approaches. [ABSTRACT FROM AUTHOR]

Recent research in the field of electrochemical biosensors equipped with peptides and nanomaterials have been categorized, reviewed, and critically analyzed. Indeed, using these innovative biosensors can revolutionize biomedical diagnostics in the future. Saving lives, time, and money in this field will be considered as some main benefits of this type of diagnosis. Here, these biosensors have been categorized and evaluated in four main sections. In the first section, the focus is on investigating the types of electrochemical peptide-based nanobiosensors applied to detect pathogenic microorganisms, microbial toxins, and viruses. In the second section, due to the importance of rapid diagnosis and prognosis of various cancers, the electrochemical peptide-based nanobiosensors designed to detect cancer biomarkers have been reviewed and analyzed. In the third section, the electrochemical peptide-based nanobiosensors, which were applied to detect the essential and effective biomolecules in the various diseases, and health control, including enzymes, hormones, biomarkers, and other biomolecules, have been considered. Finally, using a comprehensive analysis, all the used elements in these biosensors have been presented as conceptual diagrams that can effectively guide researchers in future developments. The essential factors in evaluating and analyzing these electrochemical peptide-based nanobiosensors such as analyte, peptide sequence, functional groups interacted between the peptide sequences and other biosensing components, the applied nanomaterials, diagnostic techniques, detection range, and limit of detection have also been included. Other analyzable items such as the type of used redox marker and the location of the peptide sequence against the signal transducer were also considered. [ABSTRACT FROM AUTHOR]

Some of pathogenic bacteria and fungi have the ability to produce fetal toxins which may be the direct causes of cytotoxicity or cellular dysfunction in the colonization site. Biological and non-biological environmental factors, challenge and microbes influence the effect of toxins on these pathogens. Modern research mentions that many natural materials can reduce the production of toxins in pathogenic microbes. However, researches that explain the mechanical theories of their effects are meager. This review aimed to discuss the ameliorative potential role of plant-derived compounds and probiotics to reduce the toxin production of food-borne microbes either in poultry bodies or poultry feedstuff. Moreover, studies that highlight their own toxicological mechanisms have been discussed. Adding natural additives to feed has a clear positive effect on the enzymatic and microbiological appearance of the small intestine without any adverse effect on the liver. Studies in this respect were proposed to clarify the effects of these natural additives for feed. In conclusion, it could be suggested that the incorporation of probiotics, herbal extracts, and herbs in the poultry diets has some beneficial effects on productive performance, without a positive impact on economic efficiency. In addition, the use of these natural additives in feed has a useful impact on the microbiological appearance of the small intestine and do not have any adverse impacts on intestinal absorption or liver activity as evidenced by histological examination. [ABSTRACT FROM AUTHOR]

The rapid implementation of anaerobic digestion for renewable energy production has resulted in increased generation of anaerobically digested slurry, which contains a variety of pollutants and therefore has the potential to cause serious environmental problems. Tidal flow constructed wetlands, which could generate beneficial oxygen conditions, were investigated for their success in removing nitrogen, organic matter and pathogens in anaerobically digested slurry. The results indicated that tidal operation had a positive effect on promoting NH -N and organic matter (chemical oxygen demand (COD)) removal. With an average influent NH -N concentration of 288 mg/L and COD concentration of 839 mg/L, the average removal efficiency reached up to 93% (325 g/m day) for NH -N and 53% (603 g/m day) for COD, with total inorganic nitrogen (TIN) removal efficiency of 51% (226 g/m day). The nitrogen removal in the tidal-operated CWs is highly dependent on the flooded and drained (F/D) time ratio. Changing flooded time from 3 to 5 h enhanced denitrification (nitrite reductase-K (nirK) abundance) and further resulted in improved TIN removal efficiency of 62% (237 g/m day). The removal of faecal indicators was also examined, with reduction rate of approximately 0.9 log CFU/100 mL for both Escherichia coli and total coliforms, which was independent of the influent loadings and differing flooded/drained time ratio. Tidal flow CWs were demonstrated to have the high potential to treat diluted anaerobically digested slurry. [ABSTRACT FROM AUTHOR]

Analytical techniques for detection of ochratoxin A (OTA) in food products and blood serum are of great significance. In this study, a fluorescent aptasensor was developed for sensitive and specific detection of OTA, based on a DNA pyramid nanostructure (DPN) and PicoGreen (PG) dye. The designed aptasensor inherits characteristics of DPN, such as high stability and capacity for PG loading. PG, as a fluorescent dye, could bind to double-stranded DNA (dsDNA). In the absence of OTA, the pyramid structure of DPN remains intact, leading to a very strong fluorescence emission. Because of higher affinity of aptamer for its target relative to its complementary strand, upon addition of target, the pyramid structure of DPN is disassembled, leading to a weak fluorescence emission. The presented aptasensor showed high specificity toward OTA with a limit of detection (LOD) as low as 0.135 nM. Besides, the designed sensing strategy was successfully utilized to recognize OTA in serum and grape juice with LODs of 0.184 and 0.149 nM, respectively. [ABSTRACT FROM AUTHOR]

Asparagus officinalis L. is an important crop in many European countries, likely infected by a number of Fusarium species. Most of them produce mycotoxins in plant tissues, thus affecting the physiology of the host plant. However, there is lack of information on Fusarium communities in wild asparagus, where they would definitely have considerable environmental significance. Therefore, the main scientific aim of this study was to identify the Fusarium species and quantify their typical mycotoxins present in wild asparagus plants collected at four time points of the season. Forty-four Fusarium strains of eight species- Fusarium acuminatum, Fusarium avenaceum, Fusarium culmorum, Fusarium equiseti, Fusarium oxysporum, Fusarium proliferatum, Fusarium sporotrichioides, and Fusarium tricinctum-were isolated from nine wild asparagus plants in 2013 season. It is the first report of F. sporotrichioides isolated from this particular host. Fumonisin B was the most abundant mycotoxin, and the highest concentrations of fumonisins B-B and beauvericin were found in the spears collected in May. Moniliformin and enniatins were quantified at lower concentrations. Mycotoxins synthesized by individual strains obtained from infected asparagus tissues were assessed using in vitro cultures on sterile rice grain. Most of the F. sporotrichioides strains synthesized HT-2 toxin and F. equiseti strains were found to be effective zearalenone producers. [ABSTRACT FROM AUTHOR]

New and conjugated mycotoxins of concern to regulators are frequently being identified, necessitating the costly need for new method development and sample reanalysis. In response, we developed an LC-data independent acquisition (LC-DIA) method on a Q-Exactive Orbitrap mass spectrometer tailored for mycotoxins analysis. This method combines absolute quantification of targeted fungal metabolites with non-targeted digital archiving (DA) of data on all ionizable compounds for retrospective analysis. The quantitative power of this approach was assessed by spiking 23 mycotoxins at a range of concentrations into clean maize extracts. The linearity and limits of detection achieved were comparable to conventional LC-MS/MS and significantly better than 'all-ion-fragmentation' scanning mode. This method was applied to single kernel analysis of Fusarium infected maize, where we quantified nine Fusarium metabolites and three metabolites from unexpected contaminations by Alternaria and Penicillium species. Retrospective analysis of this data set allowed us to detect the recently reported 15-acetyldeoxynivalenol-3- O-β-D-glucoside without requiring re-analysis of the samples. To our knowledge, this is the first reported occurrence of this conjugated mycotoxin in naturally contaminated maize, and led us to further study maize artificially inoculated with the 3-acetyldeoxynivalenol and 15-acetyldeoxynivalenol chemotypes of Fusarium graminearum. Analysis of these samples showed that the maize genotype tested glycosylates 15-acetyldeoxynivalenol but not 3-acetyldeoxynivalenol likely because the glycosylation site was blocked. In addition to confirming that these two F. graminearum chemotypes behave differently when infecting the host plant, it demonstrates the utility of using a single screening method to quantify known mycotoxins and archive a completely non-targeted dataset for future analysis. [ABSTRACT FROM AUTHOR]

Lead (Pb) and decabromodiphenyl ether (BDE209) are main pollutants at electric waste (e-waste) recycling sites (EWRSs), and their joint toxicological effects have received extensive attention. Frequently, soil pollution at EWRSs usually results from the occurrence of repeated single or multiple pollution events, with continuous impacts on soil microorganisms. Therefore, a laboratory incubation study was conducted to determine Pb bioavailability and microbial toxicity in repeated Pb-polluted soil in the presence of BDE209 for the first time. We evaluated the impacts of repetitive exposure trials on chemical fractions of Pb, and the results showed that repeated single Pb pollution event resulted in an increase of carbonates fraction of Pb, which was different from one-off single Pb exposure. Moreover, one-off Pb-treated groups exhibited higher I (reduced partition index) values on day 30 and all treatments remained the same I level at the end of incubation period. The parameters of microbial toxicity were well reflected by soil enzymes. During the entire incubation, the dehydrogenase and urease activities were significantly inhibited by Pb ( P < 0.01), and BDE209 supply could weaken the adverse influence. Additionally, significant correlations between available or metastable Pb and the two soil enzymes were clearly observed ( P < 0.05 or 0.01). Such observations would provide useful information for ecological effects of Pb and BDE209 at EWRSs. [ABSTRACT FROM AUTHOR]

Rapid and profound changes in earth surface environments and biota across the Permian-Triassic boundary are well known and relate to the end-Permian mass extinction event. This major crisis is demonstrated by abrupt facies change and the development of microbialite carbonates on the shallow marine shelves around Palaeo-Tethys and western Panthalassa. Microbialites have been described from a range of sites in end-Permian and basal Triassic marine sedimentary rocks, immediately following the end-Permian mass extinction. Here, we present geochemical data primarily focused on microbialites. Our geochemical analysis shows that U, V, Mo and REE (Ce anomaly) may be used as robust redox proxies so that the microbialites record the chemistry of the ancient ambient sea water. Among the three trace metals reputed to be reliable redox proxies, one (V) is correlated here with terrigenous supply, the other two elements (U and Mo) do not show any significant authigenic enrichment, thereby indicating that oxic conditions prevailed during the growth of microbialites. REE profiles show a prominent negative Ce anomaly, also showing that the shallow marine waters were oxic. Our geochemical data are consistent with the presence of some benthic organisms (ostracods, scattered microgastropods, microbrachiopods and foraminifers) in shallow marine waters that survived the mass extinction event. [ABSTRACT FROM AUTHOR]

Benthic cyanobacterial blooms are increasing worldwide and can be harmful to human and animal health if they contain toxin-producing species. Microbial interactions are important in the formation of benthic biofilms and can lead to increased dominance and/or toxin production of one or few taxa. This study investigated how microbial interactions contribute to proliferation of benthic blooms dominated by the neurotoxin-producing Phormidium autumnale. Following a rainfall event that cleared the substrate, biofilm succession was characterised at a site on the Hutt River (New Zealand) by sampling every 2-3 days over 32 days. A combination of morphological and molecular community analyses (automated ribosomal intergenic spacer analysis and Illumina™ MiSeq sequencing) identified three distinct phases of succession in both the micro-algal and bacterial communities within P. autumnale-dominated biofilms. Bacterial composition shifted between the phases, and these changes occurred several days before those of the micro-algal community. Alphaproteobacteria and Betaproteobacteria dominate in the early phase; Alphaproteobacteria, Betaproteobacteria, Sphingobacteria and Flavobacteria in the mid-phase; and Sphingobacteria and Flavobacteria in the late phase. Collectively, the results suggest that succession is driven by bacteria in the early stages but becomes dependent on micro-algae in the mid- and late stages of biofilm formation. [ABSTRACT FROM AUTHOR]

Elevated plant and microbial toxicity throughout the season was found in field experiments on bioremediation of diesel fuel (DF)-contaminated soil (4.5%). This was an indication of the presence of mobile toxic DF components and their metabolites. Introduction of granulated activated charcoal (GAC) was shown to decrease the bio- and phytotoxicity of petroleum-contaminated soil, resulting in a sharp increase in the abundance of petroleum degraders (both aboriginal and inoculated ones), as well as to create conditions for improved growth at the stage of post-treatment by phytoremediation. In spite of short-time deceleration of DF degradation, more complete decrease of hydrocarbon concentrations occurred in the presence of GAC, while simultaneous introduction of the sorbent and a biopreparation (and association of mesophilic petroleum-degrading strains) provided the best results. In these variants the concentration of petroleum hydrocarbons decreased to 0.19-0.21 and 0.13-0.14%, respectively, which was 1.5 and 2 times lower than the values for unsupplemented control. Thus, GAC introduction during bioremediation of DF-contaminated soils increases the efficiency of remediation and localizes the pollutants in the treated layer, which decreases the risk of their penetration into groundwater during in situ soil treatment. [ABSTRACT FROM AUTHOR]

This study, based within the catchment area of the River Frome, an important chalk stream in the south of England, compared ciliated protozoan communities associated with three species of aquatic macrophyte common to lotic habitats: Ranunculus penicillatus subsp. pseudofluitans, Nasturtium officinale and Sparganium emersum. A total of 77 ciliate species were counted. No species-specific ciliate assemblage was found to be typical of any one plant species. Ciliate abundance between plant species was determined to be significantly different. The ciliate communities from each plant species were unique in that the number of species increased with ciliate abundance. The community associated with R. penicillatus subsp. pseudofluitans showed the highest consistency and species richness whereas S. emersum ciliate communities were unstable. Most notably, N. officinale was associated with low ciliate abundances and an apparent reduction in biofilm formation, discussed herein in relation to the plant's production of the microbial toxin phenethyl isothiocyanate. We propose that the results reflect differences in the quantity and quality of biofilm present on the plants, which could be determined by the different plant morphologies, patterns of plant decay and herbivore defense systems, all of which suppress or promote the various conditions for biofilm growth. [ABSTRACT FROM AUTHOR]

A degrading microbial consortium highly tolerant to three-, four- and five-ring polycyclic aromatic hydrocarbons (PAHs) was selected from 50 fungal and bacterial isolates obtained from crude oil-contaminated soils. Morphological and molecular studies indicated that isolated fungi belonged to genera Aspergillus, Penicillium, Fusarium, Trichoderma, Scedosporium, and Acremonium and bacteria to Pseudomonas, Klebsiella, Bacillus, Enterobacter, Streptomyces, Stenotrophomonas, Kocuria, and Delftia genera. Individual fungal and bacterial isolates were evaluated for their potential to tolerate high concentrations of different molecular weight PAHs, as phenantrene (Phe), pyrene (Pyr), and benzo[ a]pyrene (BaP) by surface plate assays, showing significant differences in extension rates for fungi and inhibition ratios for bacteria when both were exposed to 0-6,000 mg of PAHs per liter. Trichoderma asperellum H15, Aspergillus nomius H7, Aspergillus flavus H6, Pseudomonas aeruginosa B7, Klebsiella sp. B10, and Stenotrophomonas maltophilia B14 grew using PAHs as sole carbon source and presented a remarkably high tolerance to PAHs, up to 6,000 mg l. The consortium composed of 12 fungal and bacterial PAH-tolerant isolates for the bioremediation of a PAH-contaminated soiled to a removal of 87.76 % Phe, 48.18 % Pyr, and 56.55 % BaP after 14 days. The degrading microbial consortium presented high potential for bioremediation and may be useful for the treatment of sites polluted with PAHs due to their elevated tolerance to high molecular weight (HMW) PAHs and their capacity to utilize them as energy source. This is the first study which evaluated the microbial tolerance to extreme concentrations of PAHs, resulting in a degrading consortium and highly tolerant consortium compared with those reported in other studies, where the concentrations tested are low. [ABSTRACT FROM AUTHOR]

The retrograde membrane transport pathways from endosomes to the trans-Golgi network (TGN) are now recognized as critical intracellular pathways to recycle and shuttle a selective subgroup of membrane proteins, including sorting receptors, membrane-bound enzymes, transporters, as well as providing an avenue for the intracellular transport of various bacterial toxins. Multiple pathways from endosomes to the TGN have now been defined which differ between the cargo transported and the machinery used. Here, we review advances in these pathways and the requirement for TGN organization, and also discuss the development of unbiased analytical approaches to quantitatively track cargo that use these endosome-to-TGN pathways. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the incidence of total aflatoxin (AF), ochratoxin A, T-2 toxin, deoxynivalenol (DON), zearalenone (ZEA), and fumonisin (FB) in dairy cattle, beef cattle, and lamb-calf feeds. A total of 180 dairy cattle, beef cattle, and lamb-calf feeds (60 samples each) were randomly collected from farms, feed mills, and villages in Burdur province, between September 2006 and August 2007. All samples were analyzed by the competitive Enzyme Linked Immuno Sorbent Assay (ELISA). The most frequent mycotoxin detected was total AF, which was found in 108 samples (60 %) in concentrations ranging from 3.82 to 116.83 μg kg, followed by DON that was detected in 87 samples (48.3 %), in concentrations ranging from 18.50 to 500 μg kg. Ochratoxin A (OTA), T-2 toxin, ZEA, and FB were found in 84 (46.7 %), 85 (47.2 %), 57 (31.7 %), and 19 (10.6 %) samples, respectively, in concentrations of 1.01 to 15.85 μg kg for OTA, 3.85 to 52.36 μg kg for T-2 toxin, 2.10 to 29.30 μg kg for ZEA, and 2.69 to 4.96 mg kg for FB. It was concluded that feed samples in Burdur province were contaminated by mycotoxins, and the levels of total aflatoxin in the samples were considered a risk to animal health. [ABSTRACT FROM AUTHOR]

Enterohemorrhagic (EHEC) and enteroaggregative (EAEC) are two pathotypes of diarrheagenic Escherichia coli. EAEC strains express adhesins called aggregate adherence fimbriae (AAFs) which the bacteria use to adhere to intestinal mucosa. EHEC virulence factor is Shiga toxin which belongs to the AB5 toxin family. B subunit, the nontoxic part of Shiga toxin (StxB), forms a homo pentamer and is responsible for binding to target cells. StxB has recently been proven to have adjuvant activity. In the current study we fused StxB encoding gene to 3' end of genes encoding two variants of AAFs, i.e., AAF/I and AAF/II. The in silico studies on tertiary structure and biochemical characteristics of Shiga toxin A subunit (StxA) revealed more resemblance to AAF/II than AAF/I. The constructs were prepared in a way that StxB could imitate its natural structure (pentamer formation) and its position (C-terminus) in the native toxin complex. The expression of these constructs showed the formation of AAF/II-B as a protein complex but with lower molecular mass than its expected size. In contrast, the AAF/I-B complex was not formed. Overall, the results of in silico studies and expression experiments together revealed that despite AAF/II-B expression, StxB failed to form pentamer. Therefore the observed protein complex has lower molecular mass. Since StxB is bound to AAF/II through disulfide bond, this bond prevents pentamer formation of StxB. However, due to the lack of disulfide bond between AAF/I and StxB, no protein complex is formed, thus StxB maintains its pentamer structure. [ABSTRACT FROM AUTHOR]

The sensitivities of four different kinds of bioassays to the toxicities of arsenite, chromate, cadmium, and copper were compared. The different bioassays exhibited different sensitivities, i.e., they responded to different levels of toxicity of each of the different metals. However, with the exception of the α-glucosidase enzyme activity, arsenite was the most toxic compound towards all the tested organisms, exhibiting the highest toxic effect on the seeds of Lactuca, with an EC value of 0.63 mg/L. The sensitivities of Lactuca and Raphanus were greater than the sensitivities of two other kinds of seeds tested. Therefore, these were the seeds appropriate for use in a seed germination assay. A high revertant mutagenic ratio (5:1) of Salmonella typhimurium was observed with an arsenite concentration of 0.1 μg/plate, indicative of a high possibility of mutagenicity. These different results suggested that a battery of bioassays, rather than one bioassay alone, is needed as a more accurate and better tool for the bioassessment of environmental pollutants. [ABSTRACT FROM AUTHOR]

Maize ( Zea mays) considered to be one of the most frequent crops in Croatia, is often contaminated with Fusarium mycotoxins. The aim of this study was to investigate the possible contamination of maize with T-2 toxin and fumonisins on 46 samples from different regions of Croatia. The highest concentrations of T-2 toxin and fumonisins were 210 and 25,200 ng/g, with mean values of 110 and 4,509 ng/g, respectively, pointing to maize contamination with these mycotoxins after the period of the year with extremely high rainfall. [ABSTRACT FROM AUTHOR]

The wheat rhizosphere-inhabiting nonpathogenic Fusarium sambucinum isolate FS-94 protected tomato from Fusarium wilt ( F. oxysporum f. sp. lycopersici) in laboratory experiments. Seed soaking or immersion of seedling roots in a FS-94 spore suspension prior to inoculation with the pathogen delayed the appearance of wilt symptoms and significantly reduced disease severity in plants of a susceptible tomato cultivar. Quantification of fungal ergosterol in infected tomato showed that protection against wilt agent was related to limitation of the pathogen growth in plants exposed to FS-94. Incubation of tomato seedlings in a FS-94 spore suspension for 48 or 72 h led to plant protection and increased the salicylic acid (SA) concentration in their roots, suggesting that this isolate was involved in a plant-mediated mode of action and induced resistance. Soaking tomato seeds in the spore suspension did not induce SA accumulation in seedling roots, but nevertheless resulted in a significant reduction in wilt severity when the seedlings were challenged with the pathogen. In response to pathogen attack, the SA content in susceptible seedlings grown from FS-94-treated seeds started to increase within 1 day and remained elevated for 72 h. This suggests that F. sambucinum isolate FS-94 primed a SA-dependent signaling system in tomato. [ABSTRACT FROM AUTHOR]

The global prevalence of severe Clostridium difficile infection highlights the profound clinical significance of clostridial glucosylating toxins. Virulence is dependent on the autoactivation of a toxin cysteine protease, which is promoted by the allosteric cofactor inositol hexakisphosphate (InsP6). Host mechanisms that protect against such exotoxins are poorly understood. It is increasingly appreciated that the pleiotropic functions attributed to nitric oxide (NO), including host immunity, are in large part mediated by S-nitrosylation of proteins. Here we show that C. difficile toxins are S-nitrosylated by the infected host and that S-nitrosylation attenuates virulence by inhibiting toxin self-cleavage and cell entry. Notably, InsP6- and inositol pyrophosphate (InsP7)-induced conformational changes in the toxin enabled host S-nitrosothiols to transnitrosylate the toxin catalytic cysteine, which forms part of a structurally conserved nitrosylation motif. Moreover, treatment with exogenous InsP6 enhanced the therapeutic actions of oral S-nitrosothiols in mouse models of C. difficile infection. Allostery in bacterial proteins has thus been successfully exploited in the evolutionary development of nitrosothiol-based innate immunity and may provide an avenue to new therapeutic approaches. [ABSTRACT FROM AUTHOR]

Toxic blooms of the cyanobacterium Microcystis aeruginosa affect humans and animals in inland water systems worldwide, and it has been hypothesized that the development of these blooms will increase under the future scenario of global change, considering eutrophication and temperature increase as two important consequences. The importance of genetic adaptation, chance and history on evolution of growth rate, and toxin production of M. aeruginosa was studied under these new conditions. The experiment followed the idea of 'replaying life's tape' by means of the simultaneous propagation of 15 independent isolates of three M. aeruginosa strains, which were grown under doubled nutrient concentration and temperature during c. 87 generations. Adaptation by new mutations that resulted in the enhancement of growth rate arose during propagation of derived cultures under the new environmental conditions was the main component of evolution; however, chance also contributed in a lesser extension to evolution of growth rate. Mutations were selected, displacing the wild-type ancestral genotypes. In contrast, the effect of selection on mutations affecting microcystin production was neutral. Chance and history were the pacemakers in evolution of toxin production. Although this study might be considered an oversimplification of the reality, it suggest that a future scenario of global change might lead to an increase in M. aeruginosa bloom frequency, but no predictions about the frequency of toxicity can be made. [ABSTRACT FROM AUTHOR]

Humidity is an important determinant of the mycotoxin production (DON, ZEA) by Fusarium species in the grain ears. From a landscape perspective humidity is not evenly distributed across fields. The topographically-controlled redistribution of water within a single field rather leads to spatially heterogeneous soil water content and air humidity. Therefore we hypothesized that the spatial distribution of mycotoxins is related to these topographically-controlled factors. To test this hypothesis we studied the mycotoxin concentrations at contrasting topographic relief positions, i.e. hilltops and depressions characterized by soils of different soil moisture regimes, on ten winter wheat fields in 2006 and 2007. Maize was the preceding crop and minimum tillage was practiced in the fields. The different topographic positions were associated with moderate differences in DON and ZEA concentrations in 2006, but with significant differences in 2007, with six times higher median ZEA and two times higher median DON detected at depression sites compared to the hilltops. The depression sites correspond to a higher topographic wetness index as well as redoximorphic properties in soil profiles, which empirically supports our hypothesis at least for years showing wetter conditions in sensitive time windows for Fusarium infections. [ABSTRACT FROM AUTHOR]

The article focuses on a study done on the regulation of mycotoxin biosynthesis by chemicals. It states that mycotoxins, which are highly toxic pollutants of agricultural raw materials and foodstuff, are classified with polyketides. It describes how chemicals such as fungicide tricyclazole, 1-aminoethyl phosphinic acid and phosphinic analogues of alanine, aspartate and leucine can suppress polyketide biosynthesis.

Rapid microscale toxicity tests make it possible to screen large numbers of compounds and greatly simplify toxicity identification evaluation and other effect directed chemical analyses of effluents or environmental samples. Tests using Vibrio fischeri (such as Microtox®) detect toxicants that cause non-specific narcosis, but are insensitive to other important classes of contaminants. The microbial assay for risk assessment (MARA) is a 24 h multi-species test that seeks to address this problem by using a battery of ten bacteria and a fungus. But there has been little independent evaluation of this test, and there is no published information on its sensitivity to pesticides. Here, we assess the performance of MARA using a range of toxicants including reference chemicals, fungicides and environmental samples. Mean MARA microbial toxic concentrations and ICs (20% Inhibitory concentrations) indicate the toxicant concentrations affecting the more sensitive micro-organisms, while the mean IC (50% Inhibitory concentration) was found to be the concentration that was toxic to most MARA species. For the two fungicides tested, the yeast ( Pichia anomalia) was the most sensitive of the ten MARA species, and was more sensitive than the nine other yeasts tested. The test may be particularly valuable for work with fungicides. Mean MARA ICs were comparable to values for nine other yeast species and the lowest individual ICs for each toxicant were comparable to reported ICs for Daphnia magna, Selenastrum capricornutum and Microtox® bioassays. MARA organisms exhibited more variable sensitivities, with the most sensitive organism being different for different samples, enhancing the likelihood of toxicity detection and giving a toxicity 'fingerprint' that may help identify toxicants. The test, therefore, has great potential and would be valuable for ecotoxicological testing of pollutants. [ABSTRACT FROM AUTHOR]

Mycotoxins are natural contaminants produced by a range of fungal species. Their common occurrence in food and feed poses a threat to the health of humans and animals. This threat is caused either by the direct contamination of agricultural commodities or by a “carry-over” of mycotoxins and their metabolites into animal tissues, milk, and eggs after feeding of contaminated hay or corn. As a consequence of their diverse chemical structures and varying physical properties, mycotoxins exhibit a wide range of biological effects. Individual mycotoxins can be genotoxic, mutagenic, carcinogenic, teratogenic, and oestrogenic. To protect consumer health and to reduce economic losses, surveillance and control of mycotoxins in food and feed has become a major objective for producers, regulatory authorities and researchers worldwide. However, the variety of chemical structures makes it impossible to use one single technique for mycotoxin analysis. Hence, a vast number of analytical methods has been developed and validated. The heterogeneity of food matrices combined with the demand for a fast, simultaneous and accurate determination of multiple mycotoxins creates enormous challenges for routine analysis. The most crucial issues will be discussed in this review. These are (1) the collection of representative samples, (2) the performance of classical and emerging analytical methods based on chromatographic or immunochemical techniques, (3) the validation of official methods for enforcement, and (4) the limitations and future prospects of the current methods. [ABSTRACT FROM AUTHOR]

Pore-forming toxins (PFTs) are a class of potent virulence factors that convert from a soluble form to a membrane-integrated pore. They exhibit their toxic effect either by destruction of the membrane permeability barrier or by delivery of toxic components through the pores. Among the group of bacterial PFTs are some of the most dangerous toxins, such as diphtheria and anthrax toxin. Examples of eukaryotic PFTs are perforin and the membrane-attack complex, proteins of the immune system. PFTs can be subdivided into two classes, α-PFTs and β-PFTs, depending on the suspected mode of membrane integration, either by α-helical or β-sheet elements. The only high-resolution structure of a transmembrane PFT pore is available for a β-PFT—α-haemolysin from Staphylococcus aureus. Cytolysin A (ClyA, also known as HlyE), an α-PFT, is a cytolytic α-helical toxin responsible for the haemolytic phenotype of several Escherichia coli and Salmonella enterica strains. ClyA is cytotoxic towards cultured mammalian cells, induces apoptosis of macrophages and promotes tissue pervasion. Electron microscopic reconstructions demonstrated that the soluble monomer of ClyA must undergo large conformational changes to form the transmembrane pore. Here we report the 3.3 Å crystal structure of the 400 kDa dodecameric transmembrane pore formed by ClyA. The tertiary structure of ClyA protomers in the pore is substantially different from that in the soluble monomer. The conversion involves more than half of all residues. It results in large rearrangements, up to 140 Å, of parts of the monomer, reorganization of the hydrophobic core, and transitions of β-sheets and loop regions to α-helices. The large extent of interdependent conformational changes indicates a sequential mechanism for membrane insertion and pore formation. [ABSTRACT FROM AUTHOR]

Soil pseudomonads increase their competitiveness by producing toxic secondary metabolites, which inhibit competitors and repel predators. Toxin production is regulated by cell–cell signalling and efficiently protects the bacterial population. However, cell communication is unstable, and natural populations often contain signal blind mutants displaying an altered phenotype defective in exoproduct synthesis. Such mutants are weak competitors, and we hypothesized that their fitness depends on natural communities on the exoproducts of wild-type bacteria, especially defence toxins. We established mixed populations of wild-type and signal blind, non-toxic gacS-deficient mutants of Pseudomonas fluorescens CHA0 in batch and rhizosphere systems. Bacteria were grazed by representatives of the most important bacterial predators in soil, nematodes (Caenorhabditis elegans) and protozoa (Acanthamoeba castellanii). The gacS mutants showed a negative frequency-dependent fitness and could reach up to one-third of the population, suggesting that they rely on the exoproducts of the wild-type bacteria. Both predators preferentially consumed the mutant strain, but populations with a low mutant load were resistant to predation, allowing the mutant to remain competitive at low relative density. The results suggest that signal blind Pseudomonas increase their fitness by exploiting the toxins produced by wild-type bacteria, and that predation promotes the production of bacterial defence compounds by selectively eliminating non-toxic mutants. Therefore, predators not only regulate population dynamics of soil bacteria but also structure the genetic and phenotypic constitution of bacterial communities.The ISME Journal (2009) 3, 666–674; doi:10.1038/ismej.2009.26; published online 26 March 2009 [ABSTRACT FROM AUTHOR]

The mosquito-larvicidal binary toxin from Bacillus sphaericus is composed of two polypeptides called BinA and BinB with molecular masses of approximately 42 and 51 kDa. Both components are required for full activity, with BinB acting as a specificity determinant and BinA being responsible for toxic action. To investigate the role of the selected charged residues in BinA, four mutants were generated by replacing charged amino acids with alanine (R97A, E98A, R101A, and E114A). All mutant proteins were produced at high levels and formed inclusion bodies similar to that of the wild type. Mosquito-larvicidal assays against Culex quinquefasciatus larvae revealed that the mutant R97A completely lost its activity and mutants E98A, R101A, and E114A showed significantly reduced toxicity. Intrinsic fluorescence spectroscopy analysis indicated that alanine substitutions at these positions did not alter the overall structure of the toxin. Binding of the mutants to BinB was not different from that of the wild type, suggesting that these mutations did not affect BinA-BinB interaction. Results demonstrated that R97, E98, R101, and E114 neither play a direct role in maintenance of BinA structure nor are involved in BinA-BinB interaction. Since these residues are required for full activity, they may play an important role during toxin internalization and/or toxic action of BinA inside the target cells. [ABSTRACT FROM AUTHOR]

Colinearity in gene content and order between rice and closely related cereal crops has been a powerful tool for gene identification. Using a comparative genomic approach, we have identified the rice genomic region syntenous to the region of the short arm of wheat chromosome 2D, on which quantitative trait loci (QTLs) for Fusarium head blight (FHB) resistance and for controlling accumulation of the mycotoxin deoxynivalenol (DON) are closely located. Utilizing markers known to reside near the FHB resistance QTL and data from several wheat genetic maps, we have limited the syntenous region to 6.8 Mb of the short arm of rice chromosome 4. From the 6.8-Mb sequence of rice chromosome 4, we found three putative rice genes that could have a role in detoxification of mycotoxins. DNA sequences of these putative rice genes were used in BLAST searches to identify wheat expressed sequence tags (ESTs) exhibiting significant similarity. Combined data from expression analysis and gene mapping of wheat homologues and results of analysis of DON accumulation using doubled haploid populations revealed that a putative gene for multidrug resistance-associated protein (MRP) is a possible candidate for the FHB resistance and/or DON accumulation controlling QTLs on wheat chromosome 2DS and can be used as a molecular marker to eliminate the susceptible allele when the Chinese wheat variety Sumai 3 is used as a resistance source. [ABSTRACT FROM AUTHOR]

Stress acclimating plants respond to abiotic and biotic stress by remodeling membrane fluidity and by releasing α-linolenic (18:3) from membrane lipids. The modification of membrane fluidity is mediated by changes in unsaturated fatty acid levels, a function provided in part by the regulated activity of fatty acid desaturases. Adjustment of membrane fluidity maintains an environment suitable for the function of critical integral proteins during stress. α-Linolenic acid, released from membrane lipid by regulated lipase activity, is the precursor molecule for phyto-oxylipin biosynthesis. The modulation of chloroplast oleic acid (18:1) levels is central to the normal expression of defense responses to pathogens in Arabidopsis. Oleic (18:1) and linolenic (18:2) acid levels, in part, regulate development, seed colonization, and mycotoxin production by Aspergillus spp. [ABSTRACT FROM AUTHOR]

A procedure for the analytical validation of a rapid supercritical fluid extraction amperometric screening method for controlling macrocyclic lactone mycotoxins in maize flour samples has been developed. The limit established by European legislation (0.2 mg kg−1), in reference to zearalenone (ZON) mycotoxin, was taken as the reference threshold to validate the proposed method. Natural ZON metabolites were also included in this study to characterize the final screening method. The objective was the reliable classification of samples as positive or negative samples. The cut-off level was fixed at a global concentration of mycotoxins of 0.17 mg kg−1. An expanded unreliability zone between 0.16 and 0.23 mg kg−1 characterized the screening method for classifying the samples. A set of 30 samples was used for the final demonstration of the reliability and usefulness of the method. [ABSTRACT FROM AUTHOR]

Ochratoxin A is a mycotoxin produced by several Aspergillus and some Penicillium species which may be present in food and feed products. It can be enzymatically hydrolyzed into ochratoxin α and l-β-phenylalanine, thereby decreasing its toxicity. The ochratoxin A degradation capacity of Aspergillus niger is well known and here we report the isolation and purification of a novel enzyme from A. niger that hydrolyzes this mycotoxin. A wheat germ medium supplemented with ochratoxin A was used to produce the enzyme, which was purified from culture filtrate by acetone precipitation and anion exchange chromatography. An overall purification of 2.5-fold with a recovery of 68% and a final specific activity of 36 U/mg was obtained. The enzyme is a metalloenzyme as it was inhibited at 10 mM EDTA, whereas PMSF had no effect. The ochratoxin A hydrolytic enzyme presented a V max of 0.44 μM/min and a K m of 0.5 mM when the reaction was carried out at pH 7.5 and 37°C. [ABSTRACT FROM AUTHOR]

Clathrin seems to be dispensable for some endocytic processes and, in several instances, no cytosolic coat protein complexes could be detected at sites of membrane invagination. Hence, new principles must in these cases be invoked to account for the mechanical force driving membrane shape changes. Here we show that the Gb3 (glycolipid)-binding B-subunit of bacterial Shiga toxin induces narrow tubular membrane invaginations in human and mouse cells and model membranes. In cells, tubule occurrence increases on energy depletion and inhibition of dynamin or actin functions. Our data thus demonstrate that active cellular processes are needed for tubule scission rather than tubule formation. We conclude that the B-subunit induces lipid reorganization that favours negative membrane curvature, which drives the formation of inward membrane tubules. Our findings support a model in which the lateral growth of B-subunit–Gb3 microdomains is limited by the invagination process, which itself is regulated by membrane tension. The physical principles underlying this basic cargo-induced membrane uptake may also be relevant to other internalization processes, creating a rationale for conceptualizing the perplexing diversity of endocytic routes. [ABSTRACT FROM AUTHOR]

The mammalian olfactory system mediates various responses, including aversive behaviours to spoiled foods and fear responses to predator odours. In the olfactory bulb, each glomerulus represents a single species of odorant receptor. Because a single odorant can interact with several different receptor species, the odour information received in the olfactory epithelium is converted to a topographical map of multiple glomeruli activated in distinct areas in the olfactory bulb. To study how the odour map is interpreted in the brain, we generated mutant mice in which olfactory sensory neurons in a specific area of the olfactory epithelium are ablated by targeted expression of the diphtheria toxin gene. Here we show that, in dorsal-zone-depleted mice, the dorsal domain of the olfactory bulb was devoid of glomerular structures, although second-order neurons were present in the vacant areas. The mutant mice lacked innate responses to aversive odorants, even though they were capable of detecting them and could be conditioned for aversion with the remaining glomeruli. These results indicate that, in mice, aversive information is received in the olfactory bulb by separate sets of glomeruli, those dedicated for innate and those for learned responses. [ABSTRACT FROM AUTHOR]

Milo disease in sorghum is caused by isolates of the soil-borne fungus Periconia circinata that produce PC-toxin. Susceptibility to milo disease is conditioned by a single, semi-dominant gene, termed Pc. The susceptible allele ( Pc) converts to a resistant form ( pc) spontaneously at a gametic frequency of 10−3 to 10−4. A high-density genetic map was constructed around the Pc locus using DNA markers, allowing the Pc gene to be delimited to a 0.9 cM region on the short arm of sorghum chromosome 9. Physically, the Pc-region was covered by a single BAC clone. Sequence analysis of this BAC revealed twelve gene candidates. Several of the predicted genes in the region are homologous to disease resistance loci, including one NBS-LRR resistance gene analogue that is present in multiple tandem copies. Analysis of pc isolines derived from Pc/Pc sorghum suggests that one or more members of this NBS-LRR gene family are the Pc genes that condition susceptibility. [ABSTRACT FROM AUTHOR]

Toxic contaminants may enter estuarine ecosystems through a variety of pathways. When sediment contaminant levels become sufficiently high, they may impact resident biota. One approach to predict sediment-associated toxicity in estuarine ecosystems involves the use of sediment quality guidelines (ERMs, ERLs) and site-specific contaminant chemistry while a second approach utilizes site-specific ecological sampling to assess impacts at the population or community level. The goal of this study was to utilize an integrated approach including chemical contaminant analysis, sediment quality guidelines and grass shrimp population monitoring to evaluate the impact of contaminants from industrial sources. Three impacted sites and one reference site were selected for study. Grass shrimp populations were sampled using a push-netting approach. Sediment samples were collected at each site and analyzed for metals, polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and pesticides. Contaminant levels were then compared to sediment quality guidelines. In general, grass shrimp population densities at the sites decreased as the ERM quotients increased. Grass shrimp densities were significantly reduced at the impacted site that had an ERM exceedance for chromium and the highest Mean ERM quotient. Regression analysis indicated that sediment chromium concentrations were negatively correlated with grass shrimp density. Grass shrimp size was reduced at two sites with intermediate levels of contamination. These findings support the use of both sediment quality guidelines and site-specific population monitoring to evaluate the impacts of sediment-associated contaminants in estuarine systems. [ABSTRACT FROM AUTHOR]

The occurrence of mycotoxins in barley, sorghum, teff (Eragrostis tef) and wheat from Ethiopia has been studied. Samples were analyzed for aflatoxin B1 (AFB1), ochratoxin A (OTA), deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEN) using high performance liquid chromatography (HPLC) and for fumonisins (FUM) using enzyme linked immunosorbent assay (ELISA). AFB1 and OTA were detected in samples of all the four crops. AFB1 was detected in 8.8% of the 352 samples analyzed at concentrations ranging from trace to 26 μg kg−1. OTA occurred in 24.3% of 321 samples at a mean concentration of 54.1 μg kg−1 and a maximum of 2106 μg kg−1. DON occurred in barley, sorghum and wheat at 40–2340 μg kg−1 with an overall incidence of 48.8% among the 84 mainly ‘suspect’ samples analyzed; NIV was co-analyzed with DON and was detected at 40 μg kg−1 in a wheat sample and at 50, 380, and 490 μg kg−1 in three sorghum samples. FUM and ZEN occurred only in sorghum samples with low frequencies at concentrations reaching 2117 and 32 μg kg−1, respectively. The analytical results indicate higher mycotoxin contamination in sorghum, which could be related to the widespread storage of sorghum grain in underground pits leading to elevated seed moisture contents. This is the first report on the occurrence of OTA in teff. [ABSTRACT FROM AUTHOR]

A survey was conducted at a poultry feed production unit in Kuwait for mycotoxin contamination in the samples of yellow maize, soybean meal, wheat bran used as raw material and the poultry feed prepared for broiler starter, broiler finisher, and layer mash. Individual aflatoxins were determined by high-pressure liquid chromatography after immunoaffinity column purification. Repeated analysis revealed average aflatoxin concentration in maize at 0.27 ppb (range 0 to 1.69 ppb), soybean meal at 0.20 ppb (range 0 to 1.27 ppb), wheat bran at 0.15 ppb (range 0 to 1.07 ppb), prepared poultry feed for broiler starter at 0.48 ppb (range 0 to 3.26 ppb), broiler finisher at 0.39 ppb (range 0 to 1.05 ppb), and layer mash at 0.21 ppb (range 0 to 1.30 ppb). Other mycotoxins (ochratoxin, fumonisin, deoxynivalenol (DON), and zearalenone), were detected by quantitative enzyme-linked immunosorbent assay. The average levels of ochratoxin A ranged from 4.6 to 9.6 ppb, fumonisin from 1.4 to 3.2 ppm, DON from 0.17 to 0.29 ppm, and zearalenone from 46.4 to 67.6 ppb in various commodities and prepared feed samples. The study revealed the coexistence of determined mycotoxins, although their concentrations in general were found to be lower than the permissible levels, wherever defined, for the poultry feed. [ABSTRACT FROM AUTHOR]

A new method based on the use of a two-phase separation system is proposed for the clean-up of aflatoxins after their extraction from food samples. After extraction from the sample with an appropriate solvent, aflatoxins contained in organic solvent diffuse to the aqueous phase. At that stage their relative concentration is measured fluorimetrically at 350 nm (λexc). Different organic solvents were tested, and the composition of the aqueous phase was varied. The data obtained showed that the best efficiencies were observed with toluene and PBS-65% methanol. The method was simpler and shorter, and capable of monitoring aflatoxins in samples with a LOD equal to 0.3 µg kg−1 (AFB1). However, the proposed method shows limited selectivity and could thus serve as a screening method for total aflatoxins or be combined with chromatographic techniques for individual aflatoxin determination. [ABSTRACT FROM AUTHOR]

Aspergillus flavus is a filamentous fungus that produces mycotoxins in many food and feed crops, such as maize (Zea mays L.). Isolates were analyzed for toxin production by nucleic acid profiles in an attempt to differentiate aflatoxigenic from nonaflatoxigenic isolates. A total of 41 aflatoxigenic and 34 nonalfatoxigenic isolates were included in the study. The isolates were evaluated initially using DNA amplification fingerprinting (DAF) without clear resolution of the groups. A weak association of aflatoxigenic isolates was observed, as evidenced by their clustering in 18 of 81 trees recovered from maximum parsimony analysis of binary characters derived from arbitrary signatures from amplification profiles (ASAP) data; nonaflatoxigenic isolates exhibited a pattern of paraphyletic laddering. Up to five markers unambiguously supported the aflatoxigenic isolate grouping, but the presence of alternative conflicting topologies in equally parsimonious trees precluded the observation of meaningful statistical support. With additional markers for genome of A. flavus, this method could be used to resolve toxigenic from nontoxigenic strains. This additional work could resolve aflatoxigenic isolates of A. flavus present on maize plants using ASAP, which would reduce labor intense costs and potentially lead to faster determination of resistant cultivars in breeding efforts. [ABSTRACT FROM AUTHOR]

Streptococcus pyogenes (group A Streptococcus [GAS]) is an important pathogen whose virulence is related to the production of exotoxins and the presence of particular surface components. One hundred eighty-two GAS strains were collected in northwestern Italy between 1994 and 2002 and analyzed for phenotypic characteristics (opacity factor, proteolyic activity, and antimicrobial susceptibility) and by polymerase chain reaction for the presence of genes responsible for the production of exotoxins implicated in pathogenesis speA and speF and of prtF 1 (encoding fibronectin-binding protein F1). All strains were speF positive and 19.2% were speA positive and prtF 1 negative, whereas the prtF 1 gene was identified in 39.5% of the other strains. Of these, approximately half revealed the same pulse-field gel electrophoresis (PFGE) pattern but differed in both speA gene and macrolide resistance. [ABSTRACT FROM AUTHOR]

We developed a fusion toxin, DT388IL3, consisting of the catalytic and translocation domains of diphtheria toxin (DT388) linked to interleukin 3 (IL3) for the treatment of patients with acute myeloid leukemia (AML). Our goal in this study was to estimate a range for the maximum tolerated dose (MTD) and to evaluate the dose-limiting toxicity (DLT) of DT388IL3 in cynomolgus monkeys ( Macaca fasicularis), which possess cross-reactive IL3 receptors. In our previous study, we administered up to six infusions of DT388IL3 at 40, 60, or 100 μg/kg every other day to three pairs (one male monkey and one female monkey) of young adult monkeys. In five of six monkeys, results showed a dose-dependent increase in malaise and anorexia but no consistent abnormalities in serum chemistries or blood counts. There was no evidence of organ damage by blood tests or histopathology. However, the female treated at 100 μg/kg, died of moderate to severe vasculitis of multiple tissues. Based on these findings, this study repeated the 100 μg/kg group and added a group that received 150 μg/kg in an effort to confirm a dose response. Two female monkeys were treated with up to six infusions of DT388IL3 at 100 μg/kg or 150 μg/kg every other day. One additional female monkey was treated as a negative control. Monkeys in the 100 μg/kg group showed moderate malaise and anorexia, but no consistent abnormalities in blood counts or serum chemistries. Moderate elevations of liver enzymes were noted in the 150 μg/kg group in addition to severe malaise and anorexia. No significant findings were revealed at gross necropsy. The histopathological findings revealed regenerative myeloid hyperplasia and hepatic degeneration and regeneration in the 150 μg/kg group. Similar lesions of less severity were detected in the 100 μg/kg group. DT388IL3 plasma half-life was approximately 20 min with a peak concentration of approximately 2 μg/ml (30,000 pM). The IC50 for AML blasts in vitro was 6 pM. Collectively, our results suggest that DT388IL3 can be tolerated at doses up to 100 μg/kg in a nonhuman primate, which is higher than previously reported for other AML directed diphtheria toxin fusion proteins, and should in principle allow for dose escalation with reduced toxic side effects. Based on these findings a phase I clinical trial has recently been initiated with DT388IL3 for the treatment of AML. [ABSTRACT FROM AUTHOR]

The mycotoxin ochratoxin A (OTA) is a common contaminant of many foodstuffs and, consequently, is present in a large proportion of tested populations of humans and commercial animals. The predominant effects of OTA are manifested in the kidney where the severity varies from salt wasting to renal carcinoma formation in a concentration-dependent fashion. The MDCK-C7 renal cell culture model responds to various hormones known to regulate electrolyte and fluid balance and was used as a model to study the chronic effects of an acute exposure to low dose OTA. The natriferic hormones aldosterone and insulin-like growth factor 1 (IGF1) both stimulate Na+flux in a reabsorptive direction via activation of the epithelial Na+ channel (ENaC). In contrast, anti-diuretic hormone (ADH) stimulates three separate and temporally distinct ion transport responses, one of which is Na+ reabsorption. Treatment of MDCK-C7 cells with OTA (100 nM) for 48 h selectively and irreversibly inhibits hormone-stimulated Na+ reabsorption via ENaC. This effect was retained for 48 cell passages after the removal of the toxin and mimics the OTA-induced salt-wasting that has been documented in clinical studies. These studies indicate that the effect of the toxin is genomic and therefore, likely to be long lasting in exposed animals and humans. [ABSTRACT FROM AUTHOR]

Bacterial protein toxins alter eukaryotic cellular processes and enable bacteria to successfully colonize their hosts. In recent years, there has been increased recognition that many bacterial toxins are multifunctional proteins that can have pleiotropic effects on mammalian cells and tissues. In this review, we examine a multifunctional toxin (VacA) that is produced by the bacterium Helicobacter pylori. The actions of H. pylori VacA represent a paradigm for how bacterial secreted toxins contribute to colonization and virulence in multiple ways. [ABSTRACT FROM AUTHOR]

Because the ability of the mycotoxin patulin (PAT) to cause gene mutations in mammalian cells is still ambiguous, we have studied the mutagenicity of PAT at the hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene locus in cultured Chinese hamster V79 cells with normal, depleted, and elevated glutathione (GSH) levels. PAT was more toxic to GSH-depleted cells than to normal cells and caused an increase of the intracellular GSH level in normal and GSH-depleted cells. It also caused synchronization of the cell cycle due to a temporary accumulation of cells in the G2/M phase; this G2/M arrest was more persistent in GSH-depleted than in normal cells. PAT gave rise to a clear and concentration-dependent induction of HPRT mutations at non-cytotoxic concentrations in V79 cells with normal GSH level; the lowest PAT concentration causing a significant number of mutant cells was 0.3 micromolar, and the mutagenic potency of PAT equaled that of the established mutagen 4-nitroquinoline-N-oxide. The mutagenicity of PAT was again more pronounced, by a factor of about three, in GSH-depleted V79 cells. Elevated GSH levels abolished all observed effects of PAT. These data support the notion that PAT is a mutagenic mycotoxin, in particular in cells with low GSH concentration. The ability of PAT to cause gene mutations in mammalian cells might have a bearing on its carcinogenicity. [ABSTRACT FROM AUTHOR]