On the evolution of decoys in plant immune systems

The Guard-Guardee model for plant immunity describes how resistance proteins (guards) in host cells monitor host target proteins (guardees) that are manipulated by pathogen effector proteins. A recently suggested extension of this model includes decoys, which are duplicated copies of guardee proteins, and which have the sole function to attract the effector and, when modified by the effector, trigger the plant immune response. Here we present a proof-of-principle model for the functioning of decoys in plant immunity, quantitatively developing this experimentally-derived concept. Our model links the basic cellular chemistry to the outcomes of pathogen infection and resulting fitness costs for the host. In particular, the model allows identification of conditions under which it is optimal for decoys to act as triggers for the plant immune response, and of conditions under which it is optimal for decoys to act as sinks that bind the pathogen effectors but do not trigger an immune response.Comment: 15 pages, 6 figure

Bioaffinity detection of pathogens on surfaces

The demand for improved technologies capable of rapidly detecting pathogens with high sensitivity and selectivity in complex environments continues to be a significant challenge that helps drive the development of new analytical techniques. Surface-based detection platforms are particularly attractive as multiple bioaffinity interactions between different targets and corresponding probe molecules can be monitored simultaneously in a single measurement. Furthermore, the possibilities for developing new signal transduction mechanisms alongside novel signal amplification strategies aremuchmore varied. In this article, we describe some of the latest advances in the use of surface bioaffinity detection of pathogens. Three major sections will be discussed: (i) a brief overview on the choice of probe molecules such as antibodies, proteins and aptamers specific to pathogens and surface attachment chemistries to immobilize those probes onto various substrates, (ii) highlighting examples among the current generation of surface biosensors, and (iii) exploring emerging technologies that are highly promising and likely to form the basis of the next generation of pathogenic sensors

Protective Yeasts Control V. anguillarum Pathogenicity and Modulate the Innate Immune Response of Challenged Zebrafish (Danio rerio) Larvae

Indexación: Web of ScienceWe investigated mechanisms involved in the protection of zebrafish (Danio rerio) larvae by two probiotic candidate yeasts, Debaryornyces hansenii 97 (Dh97) and Yarrowia Iypolitica 242 (YI242), against a Vibrio anguillarum challenge. We determined the effect of different yeast concentrations (10(4)-10(7) CFU/mL) to: (i) protect larvae from the challenge, (ii) reduce the in vivo pathogen concentration and (iii) modulate the innate immune response of the host. To evaluate the role of zebrafish microbiota in protection, the experiments were performed in conventionally raised and germ free larvae. In vitro co-aggregation assays were performed to determine a direct yeast-pathogen interaction. Results showed that both yeasts significantly increased the survival rate of conventionally raised larvae challenged with V. anguillarum. The concentration of yeasts in larvae tended to increase with yeast inoculum, which was more pronounced for Dh97. Better protection was observed with Dh97 at a concentration of 106 CFU/mL compared to 104 CFU/mL. In germ-free conditions V anguillarum reached higher concentrations in larvae and provoked significantly more mortality than in conventional conditions, revealing the protective role of the host microbiota. Interestingly, yeasts were equally (Dh97) or more effective (YI242) in protecting germ-free than conventionally-raised larvae, showing that protection can be exerted only by yeasts and is not necessarily related to modulation of the host microbiota. Although none of the yeasts co aggregated with V anguillarum, they were able to reduce its proliferation in conventionally raised larvae, reduce initial pathogen concentration in germ-free larvae and prevent the upregulation of key components of the inflammatory/anti-inflammatory response (il1b, tnfa, c3, mpx, and il10, respectively). These results show that protection by yeasts of zebrafish larvae challenged with V anguillarum relates to an in vivo anti-pathogen effect, the modulation of the innate immune system, and suggests that yeasts avoid the host-pathogen interaction through mechanisms independent of co-aggregation. This study shows, for the first time, the protective role of zebrafish microbiota against V. anguillarum infection, and reveals mechanisms involved in protection by two non-Saccharomyces yeasts against this pathogen.http://journal.frontiersin.org/article/10.3389/fcimb.2016.00127/ful

Characterising the role of GABA and its metabolism in the wheat pathogen Stagonospora nodorum

A reverse genetics approach was used to investigate the role of γ-aminobutyric acid metabolism in the wheat pathogenic fungus Stagonospora nodorum. The creation of mutants lacking Sdh1, the gene encoding succinic semialdehyde dehydrogenase, resulted in strains that grew poorly on γ-aminobutyric acid as a nitrogen source. The sdh1 mutants were more susceptible to reactive oxygen stress but were less affected by increased growth temperatures. Pathogenicity assays revealed that the metabolism of γ-aminobutyric acid is required for complete pathogenicity. Growth assays of the wild-type and mutant strains showed that the inclusion of γ-aminobutyric acid as a supplement in minimal media (i.e., not as a nitrogen or carbon source) resulted in restricted growth but increased sporulation. The addition of glutamate, the precursor to GABA, had no effect on either growth or sporulation. The γ-aminobutyric acid effect on sporulation was found to be dose dependent and not restricted to Stagonospora nodorum with a similar effect observed in the dothideomycete Botryosphaeria sp. The positive effect on sporulation was assayed using isomers of γ-aminobutyric acid and other metabolites known to influence asexual development in Stagonospora nodorum but no effect was observed. These data demonstrate that γ-aminobutyric acid plays an important role in Stagonospora nodorum in responding to environmental stresses while also having a positive effect on asexual development.The work was supported by Australian Research Council and Grains Research and Development Corporation

Assessing populations of the major cereal pathogens for reduced sensitivity to MBC, DMI and Strobilurin fungicides

End of project reportStudies of eyespot populations in winter wheat crops in the period 2001 to 2003 showed that the R type (Tapesia acuformis) is the dominant strain comprising 77% of isolates. Over 90% of isolates were resistant to MBC fungicides, 53% showed reduced sensitivity to prochloraz and 22% reduced sensitivity to cyprodinil. A study of winter wheat crops in February and March 2003 showed that resistance to strobilurin fungicides was widespread in Mycosphaerella graminicola (Septoria tritici) populations. Resistance was found in all but one of 21 crops sampled, at levels ranging from 9% to 84% with an average of 48%. Subsequent studies of 27 crops over the summer of 2003 showed that strobilunin resistance increased from an average of 31% before the application of the second (T2) spray to an average of 73%, three to four weeks after the application of the final (T3) spray. Strobilurin resistance in M. graminicola remained high in 2004, ranging from 50% to 100% with an average of 83% in populations in winter wheat crops sampled in March. The effect of different fungicide programmes on resistance was studied during summer 2003 at two experimental sites. Levels of strobilurin resistance in M. graminicola populations increased during the summer, in unsprayed plots and plots treated with triazole fungicides as well as in those treated with strobiurin fungicides at both sites. M. graminicola populations in winter wheat crops sampled in 2003 and 2004 were predominantly resistant to MBC-generating fungicides and sensitive to the triazole-based fungicides. Studies of the barley leaf blotch pathogen Rhynchosporium secalis showed that resistance to MBC fungicides occurred in 20% of isolates collected from crops from 2001 to 2003, but all isolates were sensitive to triazole and strobilurin fungicides

Drivers of Microbial Risk for Direct Potable Reuse and de Facto Reuse Treatment Schemes: The Impacts of Source Water Quality and Blending.

Although reclaimed water for potable applications has many potential benefits, it poses concerns for chemical and microbial risks to consumers. We present a quantitative microbial risk assessment (QMRA) Monte Carlo framework to compare a de facto water reuse scenario (treated wastewater-impacted surface water) with four hypothetical Direct Potable Reuse (DPR) scenarios for Norovirus, Cryptosporidium, and Salmonella. Consumer microbial risks of surface source water quality (impacted by 0-100% treated wastewater effluent) were assessed. Additionally, we assessed risks for different blending ratios (0-100% surface water blended into advanced-treated DPR water) when source surface water consisted of 50% wastewater effluent. De facto reuse risks exceeded the yearly 10-4 infections risk benchmark while all modeled DPR risks were significantly lower. Contamination with 1% or more wastewater effluent in the source water, and blending 1% or more wastewater-impacted surface water into the advanced-treated DPR water drove the risk closer to the 10-4 benchmark. We demonstrate that de facto reuse by itself, or as an input into DPR, drives microbial risks more so than the advanced-treated DPR water. When applied using location-specific inputs, this framework can contribute to project design and public awareness campaigns to build legitimacy for DPR

Ammonia based sanitation technology

Water-borne sanitation of toilet waste is not a viable option for the estimated 2.6 billion people that lack improved sanitation throughout the world. In Environmental Systems Analysis, source separating sewage systems have proven to be of interest, since both energy and nutrients are saved compared with conventional systems. As the urine and faecal matter contribute with the majority of nutrients to wastewater but constitute a small part of the volume, these fractions are suitable for nutrient recycling to agriculture. The potential content of pathogenic (disease causing) microorganisms makes it a necessity to sanitise the material before use as a fertiliser, especially as many pathogens are zoonotic, infecting both man and animal. The main objective of this study was to evaluate ammonia based sanitation technology for source separated urine and faeces aiming for production of safe fertilisers. To achieve this objective, the inactivation kinetics of several groups of organisms was investigated in relation to concentration of free ammonia, NH3, temperature and dry matter content. Inactivation of Ascaris suum eggs, Salmonella spp. Enterococcus spp., S. Typhimurium phage 28B, an f-specific RNA phage MS2 and a coliphage ΦX 174 was monitored in spiked human urine and faeces. Storage of urine diluted 1:0, 1:1 and 1:3 with water was studied at 4, 14, 24, and 34°C. Faecal material, source separated dry, was treated with urea at concentrations ranging from 0.5% to 2% at 14, 24, and 34°C. Faecal material with ash amendments was studied at 24 and 34°C, separately and with supplementary addition of 1% urea. Temperature was found to be a key factor for the efficiency of the ammonia based sanitation, both through synergy and by affecting transformation of ammonia into NH3. At 34°C the NH3 concentrations in urine and faecal material resulted in short decimal reduction (D) values for microorganism concentrations, except for the bacteriophage 28B, which showed little inactivation in stored faecal material. At 24°C, treatments of both urine and faeces with NH3 concentrations of 50 mM and above gave significant reductions whereas at lower concentrations (urine 1:3 and storage of faecal matter) little inactivation of bacteriophage 28B and ascaris eggs was observed. This means that urine must be collected as concentrated as possible in order to contain sufficient ammonia to reduce pathogens by storage. Treatment with urea, a 2% addition resulted in stable pH and NH3 concentrations that resulted in fast Salmonella spp. inactivation even at 4°C and 14°C, and inactivation of ascaris and the bacteriophage at temperature 24°C and above. Coverage with ash and lime during collection can give an enhanced pathogen inactivation when later treated in closed containers. Accompanying urea treatment of faeces collected with ash is possible but with a high pH (>10) in the material urea will not be degraded and thus not contribute to inactivation

Plant Growth Promotion and Biocontrol of Pythium ultimum by Saline Tolerant Trichoderma Isolates under Salinity Stress

This present study evaluates three isolates of Trichoderma as plant growth promoting or biological control agents: Trichoderma aggressivum f. sp. europaeum, Trichoderma saturnisporum, and the marine isolate obtained from Posidonia oceanica, Trichoderma longibrachiatum. The purpose is to contribute to an overall reduction in pesticide residues in the fruit and the environment and to a decrease in chemical fertilizers, the excess of which aggravates one of the most serious abiotic stresses, salinity. The tolerance of the different isolates to increasing concentrations of sodium chloride was evaluated in vitro, as well as their antagonistic capacity against Pythium ultimum. The plant growth promoting capacity and effects of Trichoderma strains on the severity of P. ultimum on melon seedlings under saline conditions were also analysed. The results reveal that the three isolates of Trichoderma, regardless of their origin, alleviate the stress produced by salinity, resulting in larger plants with an air-dry weight percentage above 80% in saline stress conditions for T. longibrachiatum, or an increase in root-dry weight close to 50% when T. aggressivum f. sp. europaeum was applied. Likewise, the three isolates showed antagonistic activity against P. ultimum, reducing the incidence of the disease, with the highest response found for T. longibrachiatum. Biological control of P. ultimum by T. aggressivum f. sp. europaeum and T. saturnisporum is reported for the first time, reducing disease severity by 62.96% and 51.85%, respectively. This is the first description of T. aggressivum f. sp. europaeum as a biological control agent and growth promoter. The application of these isolates can be of enormous benefit to horticultural crops, in both seedbeds and greenhouses

Reviewing factors affecting the effectiveness of decentralised domestic wastewater treatment systems for phosphorus and pathogen removal

Environmental pollution and risks to human health can result from diffuse sources of pollution originating from decentralised wastewater treatment systems (DWTS). In particular phosphorus pollution can lead to eutrophication and the downgrading of the quality of water bodies, for example, under the Water Framework Directive (WFD) in the EU, and pathogen pollution can result in increased risks of human exposure to pathogens and impacts on industries such as shellfish growing and tourism. The study reported in this paper reviews the effectiveness of various DWTS in removing phosphorus and pathogens from onsite systems. It was found that DWTS are typically not designed to specifically treat these pollutants and the most common type of DWTS, septic tanks, provide only basic treatment. Additional treatment such as filtration-based or wetland systems must be used to achieve desired levels of treatments. The performance of these systems is affected by site specific conditions, such as input load and sources, and climatic conditions and as such operational characteristics and treatment measures must be designed to take account of these factors