Recovery of Francisella tularensis from soil samples by filtration and detection by real-time PCR and cELISA

The aim of this study was to develop a specific and highly sensitive method able to detect very low concentrations of Francisella tularensis in soil samples by real-time PCR (qPCR) with SYBR Green I. tul4 gene, which encodes the 17-kDa protein (TUL4) in F. tularensis strains, was amplified using a LightCycler (LC) device. We achieved a detection limit of 0.69 fg of genomic DNA from F. tularensis subp. holarctica live vaccine strain (LVS), corresponding to a value less than 3.4 genome equivalents per reaction. The qPCR was shown to be specific, highly sensitive and reproducible. In addition, we evaluated 2 new methods for recovering bacteria from soil based on 1-step filtration using glass fiber filters and PVDF filters. These filtration methods enabled us to recover F. tularensis efficiently from soil samples. As few as 50 CFU per 0.5 g of soil were detected by qPCR. Capture enzyme-linked immunosorbent assay (cELISA) allowed us to detect and quantify the amount of bacteria recovered from soil by an immunological method. Although qPCR was more sensitive than cELISA, we did not observe substantial differences in the amount of bacteria quantified by both methods.     

Distribution of endocrine disruptors in the Mondego River estuary, Portugal

Recent studies in the Mondego River estuary demonstrated signs of pollution in the area, but the nature of the contamination remains unexplored. Because there seems to be any studies in that zone checking for the simultaneous presence of xenoestrogenic endocrine disrupting compounds, either of animal (estradiol and estrone), vegetal (daidzein, genistein and biochanin A), pharmaceutical (17alpha-ethynylestradiol) or industrial (bisphenol A, 4-octylphenol, 4-nonylphenol) origins, the main objective of this study was to investigate their presence, in every year season and at eight points along the estuarine gradient. For this propose, water samples (2 L) were collected in high and low tides, preconcentrated in the Oasis HLB cartridges and cleaned in silica cartridges before their analysis by HPLC-DAD and GC-MS. The current data showed the absence of contamination by animal or pharmaceutical estrogens and by alkylphenols, but demonstrated the presence of high levels of phytoestrogens (up to 1.1 mug/L) and of bisphenol A (up to 880.0 ng/L). Because these two chemicals existed in high environmental concentrations in the most upstream sampling station, and taking in account that these pollutants may additively contribute to endocrine disruption, namely on fish, it was concluded that at least this particularly area of the Mondego estuary deserves continuous monitoring programmes.     

Assessment of human exposure to benzene through foods from the Belgian market

Benzene is a volatile organic compound known to be carcinogenic to humans (Group 1) and may be present in food. In the present study, 455 food samples from the Belgian market were analyzed for benzene contents and some possible sources of its occurrence in the foodstuffs were evaluated. Benzene was found above the level of detection in 58% of analyzed samples with the highest contents found in processed foods such as smoked and canned fish, and foods which contained these as ingredients (up to 76.21 μg kg⁻¹). Unprocessed foods such as raw meat, fish, and eggs contained much lower concentrations of benzene. Using the benzene concentrations in food, a quantitative dietary exposure assessment of benzene intake was conducted on a national representative sample of the Belgian population over 15 years of age. The mean benzene intake for all foods was 0.020 μg kg bw d⁻¹ according to a probabilistic analysis. These values are below the minimum risk level for oral chronic exposure to benzene (0.5 μg kg bw d⁻¹).     

A method for the analysis of organophosphorus pesticide residues in Mexican axolotl

A method based on matrix solid-phase dispersion (MSPD) was developed for quantitative extraction of three organophosphorus pesticides (OPPs) from the Mexican axolotl, Ambystoma mexicanum. The determination was carried out using high- performance liquid chromatography (HPLC) with diode array spectrophotometric UV detection (DAD). The MSPD extraction with octadecylsilyl (C18) sorbent combined with a silica gel clean-up and acetonitrile elution was optimised for chlorpyrifos, fenthion and methyl parathion. The method was validated, yielding recovery values higher than 90%. The precision, expressed as the relative standard deviation (RSD), was less than or equal to 6% in muscle samples at spiking levels of 10 and 5 ppm. Linearity was studied from 15 to 60 ppm for chlorpyrifos and fenthion, and from 7.5 to 30 ppm for methyl parathion. The limits of detection (LODs) were found to be less than or equal to 0.5 ppm_.This method was applied to the analysis of samples from a chlorpyrifos-exposed axolotl, demonstrating its use as an analytical tool for toxicological studies.     

Effect of buffer strips and soil texture on runoff losses of flufenacet and isoxaflutole from maize fields

The influence of buffer strips and soil texture on runoff of flufenacet and isoxaflutole was studied for two years in Northern Italy. The efficacy of buffer strips was evaluated on six plots characterized by different soil textures; two plots had Riva soil (18.6% sand, 63.1% silt, 18.3% clay) while the remaining four plots had Tetto Frati (TF) soil (37.1% sand, 57% silt, 5.9% clay). Additionally, the width of the buffer strips, constituted of spontaneous vegetation grown after crop sowing, was also compared for their ability to abate runoff waters. Chemical residues in water following runoff events were investigated, as well as their dissipation in the soil. After the first runoff events, concentrations of herbicides in water samples collected from Riva plots were as much as four times lower in waters from TF plots. On average of two growing seasons, the field half-life of flufenacet in the upper soil layer (5 cm) ranged between 8.1 and 12.8 days in Riva soil, 8.5 and 9.3 days in TF soil. Isoxaflutole field half-life was less than 1 day. The buffer strip was very affective by the uniformity of the vegetative cover, particularly, at the beginning of the season. In TF plots, concentration differences were generally due to the presence or absence of the buffer strip, regardless of its width.     

Fate of atrazine in a soil under different agronomic management practices

Agricultural management affects the movement of atrazine in soil and leaching to groundwater. The objective of this study was to determine atrazine adsorption in a soil after 20 years of atrazine application under agronomic management practices differing in tillage practice (conventional and zero tillage), residue management (with and without residue retention) and crop rotation (wheat-maize rotation and maize monoculture). Atrazine sorption was determined using batch and column experiments. In the batch experiment, the highest distribution coefficient K_d (1.1 L kg^?1) at 0–10 cm soil depth was observed under zero tillage, crop rotation and residue retention (conservation agriculture). The key factor in adsorption was soil organic matter content and type. This was confirmed in the column experiment, in which the highest K_d values were observed in treatments with residue retention, under either zero or conventional tillage (0.81 and 0.68 L kg^?1, respectively). Under zero tillage, the fact that there was no soil movement helped to increase the K_d. The increased soil organic matter content with conservation agriculture may be more important than preferential flow due to higher pore connectivity in the same system. The soil's capacity to adsorb 2-hydroxyatrazine (HA), an important atrazine metabolite, was more important than its capacity to adsorb atrazine, and was similar under all four management practices (K_d ranged from 30 to 40 L kg^?1). The HA adsorption was attributed to the type and amount of clay in the soil, which is unaffected by agronomic management. Soils under conservation agriculture had higher atrazine retention potential than soils under conventional tillage, the system that predominates in the study area.     

The influence of environmental factors on the molluscicidal activity of Euphorbia milii latex

Abstract

The present study was undertaken to investigate the influence of biotic (snail size and presence of food during the test) and abiotic factors (temperature, water hardness and concentration of organic materials) on the molluscicidal activity of Euphorbia milii latex. Bioassays were conducted with B. glabrata (10 snails per concentration) and snail lethality was evaluated after 24 hr and 48 hr of exposure to lyophilized latex solutions. Neither the degree of water hardness, nor the presence of food during the test affected latex‐induced snail mortality. The snail size had a minor influence on E.milii‐induced snail lethality. Newly‐hatched (shell diameter ≤ 1mm) as well as young (3–8 mm) snails were slightly less susceptible than older (10–25 mm) mollusks. On the other hand, the molluscicidal effect of E.milii latex was modified by environmental factors such as temperature (i.e. LC₅₀ and LC₉₀ values were halved for every 10°C rise in temperature) and concentration of organic materials in the water (i.e. the higher the concentration of organic matter, the higher the LC₅₀ value). The efficacy of E.milii latex as a molluscicide can be modified by factors such as water temperature and concentration of organic materials, and to a lesser extent by snail size.     

Reconstruction of the environmental evolution of a Sicilian saltmarsh (Italy)

The present study deals with the reconstruction of the environmental evolution of a Trapani saltmarsh (southwestern Sicily, Italy) by combining different analytical approaches such as metal content evaluation, low-field nuclear magnetic resonance (NMR) relaxometry, and benthic foraminifera identification. A 41 cm core was collected in the sediments of a Trapani saltmarsh (southwestern Sicily, Italy) at a water depth of about 50 cm. Different time intervals were recognized, each characterized by peculiar features that testify different environmental conditions. In particular, the bottom layers of the sediment core (41–28 cm) comprised the lowest amount of mud fraction, only some selected metals, and the lowest foraminiferal density. Here, co-occurrence of abundant microcrystals of gypsum and Ammonia tepida is indicative of hyper-saline conditions. In the sediments from 28 to 6 cm, mud fraction and number of metal elements resulted higher due to the increase of the anthropogenic pressure. The sediments in the last time interval, corresponding to the environmental recovery of the saltmarsh, showed an increase of foraminiferal density, a decrease of the mud fraction, and a trend in the metal concentration attributable to the protection policy applied since 1990. NMR relaxometry parameters highlighted the changes of sediment chemical–physical heterogeneity going from the bottom to the top of the core. These heterogeneities have been related to the different intervals recognized as aforementioned. The present study highlights how the anthropogenic pressure modifies the environmental conditions of a transitional ecosystem like saltmarshes.