Assessment of molecular detection of Fusarium circinatum in insects and passive spore traps in Pinus radiata plantations

Fusarium circinatum is the causal agent of pitch canker, a destructive disease that threatens natural and planted pine forests around the world. Although pitch canker has caused problems in Spain and Portugal, concerning Europe as a whole, the fungus is not established across the pine distribution area. Its dispersion by wind and/or insect vectors could nevertheless play a role in the colonization of currently uninfected stands. It is therefore crucial to develop monitoring tools for its detection. To this end, we assessed the molecular detection of the pathogen in environmental samples of bark beetles and passive spore traps, collected in two infected Pinus radiata plantations in Basque country, Spain. The spread pattern of F. circinatum was assessed by an experimental design that included insect and spore traps installed at the centre, at the edge and outside the plots. Our results showed that F. circinatum was detected in both types of samples, at almost all collection dates. In both type of samples, positive detections were mainly found at the centre of the plots, a lower proportion at the edge, and very few outside. This suggests that long‐distance dispersion of Fusarium circinatum does not rely on wind spore dispersal neither on insect flight. Our study also shows that molecular methods are a powerful tool to monitor the pathogen in environmental samples.     

Validation and accreditation of a duplex real‐time PCR test for reliable in planta detection of Chalara fraxinea1

This work reports the optimization and provides statistically supported validation data for a real‐time detection tool targeting the emerging fungus Chalara fraxinea. The test proved to be specific and inclusive, based on computer simulation and in vitro assessments, and could detect as little as 20 fg of C. fraxinea DNA. The duplex real‐time polymerase chain reaction assay proved to be significantly more efficient and rapid than isolation on agar plates. To monitor the performance of the whole analysis process and to ensure reliability of the results, a set of controls was used systematically during the assays. A diagnosis flow diagram is proposed. The Laboratoire National de la Protection des végétaux was recently accredited by the French accreditation body for this protocol.     

Assessment of the Contamination of Cultivated Soils by Eighteen Trace Elements Around Smelters in the North of France

Ag, As, Bi, Cd, Co, Cr, Cu, Hg, In, Ni, Pb, Sb, Se, Sn,Tl, Th, U and Zn contamination of cultivated surfacehorizons has been assessed around two lead and zincsmelters in the North of France. The verticaldistribution of Ag, As, Bi, Cu, Hg, Se, Sb and Tl inthe soils has also been examined. The soils around thelead and zinc smelter at Noyelles-Godault arecontaminated by Ag, As, Bi, Cd, Cu, Hg, In, Ni, Pb, Sb,Se, Sn, Tl and Zn. The original concentration in themost contaminated soils may be multiplied by a factorof around 2 to 100, according to the element. Cadmium,Pb and Zn are the most abundant contaminants. The Pband Zn concentrations are correlated to those of theother contaminants, with the exception of Se. Aroundthe Auby zinc smelter, there is a contamination by thesame elements, but in different proportions, inaddition to Cr contamination. Contamination by Se canreach a depth of around 1 m, whereas contamination byAg, As, Bi, Cu, Hg, Sb and Tl is confined to the top 30cm. Although the contaminant content in most soilsdepends on the distance from the plant, Secontamination would appear to vary to a greater extentaccording to the physico-chemical soil conditions.     

Trace element distributions in soils developed in loess deposits from northern France

A pedo‐geochemical survey was carried out in the Nord‐Pas de Calais region (France) on soils developed in loess deposits. Total concentrations of Al, Fe and 18 trace elements, as well as common soil characteristics, were determined in samples from 52 surface and 97 deep horizons developed in these loess deposits. The Pb isotopic composition was determined in two sola. The composition of deep horizons, compared with that of the upper continental crust, with that of horizons developed from 21 other sedimentary rocks from the region and with that of loess from various parts of the world, confirms that loess from the Nord‐Pas de Calais region derives from multi‐recycled and well‐mixed ancient sedimentary rocks. Correlation analysis shows that least mobile (i.e. ionic potential (Z/r) is between 3 and 7) geogenic elements (Bi, Co, Cr, Cu, In, Ni, Pb, Sn, Tl, V, Zn) are associated with the fraction <2 µm (which we define as ‘lutum’). More mobile elements (As, Cd, Hg, Mn, Mo, Sb, Se) are less associated with this fraction. Cadmium is particularly linked to Mn. The distribution of [trace element]/([Al] or [Fe]) in the French loess gives the background content for soils developed from most sedimentary materials in northwestern Europe. Topsoils are enriched with all the trace elements examined, except Co, Cr and Ni. Enrichments with Cd, Cu, Mn and Zn are greater in cultivated soils than in forest soils. Enrichments with Pb and with Cu, Hg, Mo, Sb, Se and Sn are mainly due to human contamination through atmospheric fallout. Organic matter seems to act as a sink for all the exogenous trace elements.     

Contamination of Urban Soils in an Area of Northern France Polluted by Dust Emissions of Two Smelters

The contamination of 27 urban topsoils has been assessed around two lead and zinc smelters (Metaleurop Nord and Umicore) in the North of France. Eighteen trace elements have been analysed (Ag, As, Bi, Cd, Co, Cr, Cu, Hg, In, Ni, Pb, Sb, Se, Sn, Tl, Th, U and Zn). The investigation included the study of the vertical distribution of Cd, Pb and Zn as indicators of pollution. It was shown that Cd, In, Pb, Sb and Zn were major pollutants followed in lesser quantities by Ag, Bi, Cu and Hg. In addition, As, Ni, Se, Sn and Tl were present at levels slightly higher than regional agricultural values. The other elements (Co, Cr, Th and U) were at endogenous levels. The observations have highlighted the strong heterogeneity of the physico-chemical parameters of urban soils and the existence of heavy contamination of the under layers by Cd, Pb and Zn. A potential transfer of metals from the topsoil to the deeper layers and especially Cd and Zn, is not excluded. Indeed the soil rework is not the only factor explaining contamination level of the deeper layers of the studied soils. The comparison of the studied element concentrations in urban soils with nearby local agricultural values shows that the dust emission originating from the Metaleurop and Umicore smelters were not the only source of contamination. Thus a large contamination of the studied urban soils by Sb and In could be explained by domestic combustion of coal for heating.     

Test performance study of diagnostic procedures for identification and detection of Gibberella circinata in pine seeds in the framework of a EUPHRESCO project

Gibberella circinata (anamorph Fusarium circinatum) is a harmful fungus infecting pine trees that has been subject to European Union emergency phytosanitary measures since 2007. This fungus is spread over long distances primarily via pine seeds, consequently reliable detection tools and methods are needed to test pine seed lots imported and marketed in the EU. Several detection methods are available in the scientific literature but very limited validation data is available. For that purpose, one European EUPHRESCO project called ‘Gibcir‐Diagseed’ was launched, involving 10 countries. This project aimed to select and validate one or more diagnostic procedures targeting this pest via inter‐laboratory tests, as well as a suitable sampling method. Performance criteria were calculated for three procedures chosen by participants: isolation followed by morphological characterization, isolation followed by conventional PCR and biological enrichment followed by real‐time PCR. The results showed that all three procedures provided acceptable performance criteria but also demonstrated that none of them could generate 100% of correct results. Regarding the sampling issue, the National Plant Protection Organizations consultation enabled an agreement to be reached on the number of seeds to be tested: 400 per sample, for a 95% confidence level that an infection of at least 1% will be detected.     

Heavy Metals in Soil, Crops and Grass as a Source of Human Exposure in the Former Mining Areas (6 pp)

Background, Aim and Scope   In the former mining area of Northern France, the number of diseases (cancers, malformations, backwardness...) is about ten times above the national average. Environmental surveys conducted by the Ministry of Health have showed that more than 10% of children living around a lead smelter located in this area had lead levels higher than 100 μg L-1 of blood (25% in the closest city). Two main factors contribute to the population exposure: the ingestion of plants produced in the contaminated area, and also the ingestion or the inhalation of contaminated dust or soil. It is usually known that these particles are key routes of exposure to lead for younger children, in particular via hand-to-mouth transfer. For a better understanding of this problem, researchers investigated different exposure sources like soil and vegetable contamination. Materials and Methods: All these parameters that contribute to the assessment of environmental and health risks of metal contamination have been measured. About 170 composite samples of soils around the smelter or from reference areas have been analysed, mainly from fields (70), kitchen gardens (60) and lawns (38). Total contents of Cd and Pd were measured in the organo-mineral layer (0-25cm) of soils. Composite grass samples were also taken on the lawns. Crops and vegetables were sampled from fields and kitchen gardens and then, parts of the plant intended only for consumption were washed and analysed. Results: For the organo-mineral horizon of the studied sites and located in various environmental contexts (contaminated or reference), we found a very broad range of metal concentrations. Generally, the data showed a strong variability of the physicochemical parameters of the urban soils (kitchen gardens, lawns), in particular with regard to lead. This work showed that cultivated plants (agricultural or kitchen garden) or lawn grass are also significantly contaminated by heavy metals, especially lawn grasses, cereal grains (wheat, barley) and lettuces sampled around the smelter. Moreover, the proportion of lead present on lawn grasses could reach 50% of the total contamination of the plants because of the deposition of contaminated dust. Discussion: The values of pollution in agricultural field and kitchen garden soils located near the smelter were respectively about 20 and 30 times above the agricultural regional reference values, probably due to the atmospheric emissions from a smelter that significantly increases the concentrations of metals in the upper horizons of the neighbouring soils. This pollution results in a high level of contamination of plants grown of these soils. Conclusions: The results showed that heavy metals in soils, particles (dust and soil) or plants of kitchen gardens, lawns or playgrounds could be potentially transferable to the users, and in particular to young children, and could then contribute, to a considerable share, to the exposure of the population living in a contaminated area around a smelter brought about through inhalation, direct ingestion of particles or consumption of plants. Recommendations and Perspectives: These results highlight that the consumption of home-grown vegetables can constitute a risk of exposure, in particular for cadmium, and especially in children. This work also underlined the role of the contaminated soil particles in the Pb contamination of the human food chains and their environment. Because of the complexity of the various methods of population exposure, it will be necessary to complete the data base, in partucular in urban areas. The main objective of this future work will be to relate the degree of environmental contamination with the lead level in child bood, and to integrate other environmental compartments like outdoor and indoor dust of the dwellings into the model of pathway exposure.     

Investigation of heavy metal concentrations on urban soils,dust and vegetables nearby a former smelter site in Mortagne du Nord,Northern France

Environmental situations near former industrial sites like Mortagne du Nord (Northern France) in which the soil, dust and homegrown vegetables have been contaminated by heavy metals can represent a sanitary risk to the surrounding population. In order to evaluate the contamination level in different exposition media in this site, 26 urban topsoils, 17 dust and 38 vegetable samples were taken near the former zinc smelter site. All the observed levels on urban soil samples (except one) are over regional agricultural reference values for Cd and Pb. 45 % of the vegetable samples are over the European foodstuff limits and one dust sample taken in the school playground outdoor tarmac area exceeds the French limit in dust set up at 1,000 μg m^?2 for Pb. The Cd and Pb levels fluctuate depending on the use of the ground (kitchen garden, lawn, courtyard) and its location. The lack of simple correlations between the distance from the former smelter site and the Pb concentrations on sampled soils let us think that human activities contributed in the local urban contamination. In this context, who is now responsible for the soil contamination and the human exposure?     

How Raw and Gypsum Modified Bauxite Residues Affect Seed Germination,Enzyme Activities,and Root Development of Sinapis alba

Water, Air, & Soil Pollution - Isotopic effects of carbonyls were investigated using the simulation reactions of carbonyls with the cysteamine derivatization. Furthermore, variations of carbon...