The fate of pathogens in treated wastewater-soil-crops continuum and the effect of physical barriers.

Secondary treated wastewater (TWW) could provide a cheap and sustainable alternative to potable water (PW) irrigation and ensure food security, especially in arid and semi-arid regions. However, TWW may pose a health risk by introducing pathogens to the irrigated soil and crop, and especially to irrigated vegetables that are eaten raw. To avoid contamination, national and international authorities have mandated the use of physical barriers, such as drip irrigation and plastic mulch, to separate the irrigation water and the crops. Although the barriers are mandated, it is not clear whether they prevent contamination of crops. To evaluate the role of barriers on crop safety, cucumbers and melons were cultivated in a field irrigated with TWW or PW with the application of barriers including surface and subsurface drip irrigation and plastic mulch. Over 500 samples of water, soil and the model crops (surface and tissue) were collected during two growing seasons and used to monitor fecal indicator bacteria and pathogens using culture dependent and independent methods. The results showed that there were no statistically significant differences in both the fecal indicator and the pathogen abundance between treatments in either the soil or the crops, regardless of the water quality or barrier applied, even though TWW supported higher diversity and abundance of indicators and pathogens than PW. Moreover, the microbial communities detected in the irrigated soils and crops could not be linked to the irrigation water. The obtained results suggest that irrigation with TWW does not result in fecal pathogen contamination of the irrigated soil or crops.

Importance of soil texture to the fate of pathogens introduced by irrigation with treated wastewater.

World-wide water scarcity is urging the use of treated wastewater (TWW) for irrigation but this practice may have adverse effects on soil and crop contamination due to the introduction of potential microbial pathogens. The objective of this study was to evaluate the potential health risks caused by TWW irrigation of soils differing in their texture, i.e., soil particle fractions including sand, silt and clay. We predicted that the presence of fecal indicator bacteria (FIB) and pathogens would not be linked to TWW irrigation, yet their abundance would be favored by the smallest soil fraction (~2 nm, e.g., clay) as it provides the largest surface area. To test our hypotheses, culture dependent and independent techniques were used to monitor the presence, abundance and source of FIB and microbial pathogens (bacteria and protists) in water (TWW and potable water) and three irrigated soil types (clay, loam and loamy-sand) in a field study spanning two years. The results showed that FIB and pathogens' abundance were significantly different between water types, yet these differences did not carry to the irrigated soils. The abundance and presence of FIB and potential opportunistic or obligate human pathogens did not significantly differ (p > 0.05) between TWW and potable water irrigated soils. Moreover, the source of the FIB and potential pathogens could not be linked to irrigation with TWW. Yet, soil type significantly altered the potential pathogens' diversity (p < 0.05) and abundance (p < 0.05), and differences were affected by clay content, as predicted. The results gave no indication for potential adverse health effects associated with the application of TWW but demonstrated that clay has a particular stabilizing effect on the potential presence of microbial pathogens.

Enhanced Host-Parasite Resistance Based on Down-Regulation of Phelipanche aegyptiaca Target Genes Is Likely by Mobile Small RNA.

RNA silencing refers to diverse mechanisms that control gene expression at transcriptional and post-transcriptional levels which can also be used in parasitic pathogens of plants that Broomrapes (Orobanche/Phelipanche spp.) are holoparasitic plants that subsist on the roots of a variety of agricultural crops and cause severe negative effects on the yield and yield quality of those crops. Effective methods for controlling parasitic weeds are scarce, with only a few known cases of genetic resistance. In the current study, we suggest an improved strategy for the control of parasitic weeds based on trans-specific gene-silencing of three parasite genes at once. We used two strategies to express dsRNA containing selected sequences of three Phelipanche aegyptiaca genes PaACS, PaM6PR, and PaPrx1 (pma): transient expression using Tobacco rattle virus (TRV:pma) as a virus-induced gene-silencing vector and stable expression in transgenic tomato Solanum lycopersicum (Mill.) plants harboring a hairpin construct (pBINPLUS35:pma). siRNA-mediated transgene-silencing (20-24 nt) was detected in the host plants. Our results demonstrate that the quantities of PaACS and PaM6PR transcripts from P. aegyptiaca tubercles grown on transgenic tomato or on TRV-infected Nicotiana benthamiana plants were significantly reduced. However, only partial reductions in the quantity of PaPrx1 transcripts were observed in the parasite tubercles grown on tomato and on N. benthamiana plants. Concomitant with the suppression of the target genes, there were significant decreases in the number and weight of the parasite tubercles that grew on the host plants, in both the transient and the stable experimental systems. The results of the work carried out using both strategies point to the movement of mobile exogenous siRNA from the host to the parasite, leading to the impaired expression of essential parasite target genes.

Vetiver (Vetiveria zizanioides) responses to fertilization and salinity under irrigation conditions.

Vetiver (Vetiveria zizanioides) has not been widely introduced in arid and semi-arid regions where irrigation, fertilization, and salinity are important factors in plant growth. The main objective of this study was to determine the response of vetiver to fertilization (fertigation) and salinity and their interactions under irrigated conditions. The experiment was conducted in a greenhouse in 10-L pots. Combined effects of three nutrients concentrations and three salinity levels of electrical conductivity (EC) 1, 3 and 6 dS/m in the irrigation water on growth and transpiration of vetiver plants and the content of different elements in their foliage were studied. Similar contents of approximately 3.7 g/kg Na, approximately 5.77 g/kg Ca and approximately 2.55 g/kg Mg were found in the foliage of all the plants irrigated with the different fertilizer and salinity levels. Concentrations of 59 mg/L N and 36.1mg/L K in the irrigation water were sufficient for vetiver plants needs at the different salinity levels tested. The salinity threshold (the maximum EC in the soil solution that does not cause a significant yield reduction) for vetiver was between 3 and 6 dS/m. A concentration of 15.2mg/L P in the irrigation water was the optimum value for vetiver growth in the three salinity levels, resulting in an average content of 5.95 g/kg P in plant foliage. It is suggested that vetiver is sensitive to excess P (>8.66 g/kg). Increasing EC in the irrigation water to 6 dS/m decreased plant foliage biomass mainly due to an increase in the osmotic potential of the irrigation water and high Cl(-) concentration in the foliage.