European scale application of atmospheric reactive nitrogen measurements in a low-cost approach to infer dry deposition fluxes

A European scale network was established in 2006 as part of the NitroEurope Integrated Project to infer reactive nitrogen (Nr) dry deposition fluxes, based on low-cost sampling of gaseous and aerosol species and inferential modelling. The network provides monthly measurements of NH₃, NH₄⁺, HNO₃ and NO₃⁻, as well as SO₂, SO₄²⁻, HCl, Cl⁻ and base cations at 58 sites. Measurements are made with an established low-cost denuder methodology (DELTA) as a basis to: (1) examine temporal trends and spatial patterns across Europe, (2) improve and calibrate inferential modelling techniques to estimate exchange of Nr species, (3) provide best estimates of atmospheric dry N deposition, and (4) permit an analysis of net GHG exchange in relation to atmospheric and agricultural N inputs at the European scale. Responsibility for measurements is shared among seven European laboratories. An inter-comparison of the DELTA implementation by 6 laboratories at 4 test sites (Montelibretti, Italy; Braunschweig, Germany; Paterna, Spain and Auchencorth, UK) from July to October 2006 provided training for the laboratories and showed that good agreement was achieved in different climatic conditions (87% of laboratory site-means within 20% of the inter-laboratory median). Results obtained from the first year of measurements show substantial spatial variability in atmospheric Nr concentrations, illustrating the major local (NH₃) and regional (HNO₃, NO₃⁻, and NH₄⁺) differences in Nr concentrations. These results provide the basis to develop future estimates of site-based Nr dry deposition fluxes across Europe, and highlight the role of NH₃, largely of agricultural origin, which was the largest single constituent and will dominate dry Nr fluxes at most sites.     

Litter decomposition and nitrogen release in a sloping Mediterranean subtropical agroecosystem on the coast of Granada (SE, Spain): Effects of floristic and topographic alteration on the slope

On the coast of Granada (SE, Spain), an economically important area for subtropical fruit cultivation, the crops are grown on orchard terraces. Also, high amounts of fertilizers, often excessive, are used in this type of intensive agriculture. However, each year significant fractions of nutrients taken up by the trees return to the soil by fallen leaves. Using a litter-bag technique, we assessed the decomposition rates and N-release in various types of litter. Our main purpose was to compare two different agroecosystem scenarios: (1) an unaltered slope consisting mainly of a mixture of herbaceous plants (Papaver rhoeas, Convolvulus sp., Malva sylvestris, Reseda phyteuma, Anacyclus sp., Sinapis arvensis, Medicago sp.) among spontaneous perennial woody shrubs (Genista umbellata, Olea europaea, Lavandula officinalis, Phlomis purpurea, Retama sphaerocarpa), and (2) an altered slope cultivated with subtropical trees on terraces: loquat (Eriobotrya japonica), mango (Mangifera indica), avocado (Persea americana), and cherimoya (Annona cherimola), with groundcover plantings of aromatic, medicinal, and melliferous plants (AMMPs) on the taluses of the terraces, which are usually used for erosion control: Lavandula dentata, Thymus mastichina, Satureja obovata, Rosmarinus officinalis, Anthyllis cytisoides. In the leaves from the subtropical crops, we found the highest decomposition rates in cherimoya and the lowest in mango (1.30 and 0.64 years⁻¹, respectively). Leaves from mango and loquat registered initial peaks of N immobilization and later N-release, which was highest in cherimoya and avocado leaves (71.2 and 56.8% of the initial remaining N). In the spontaneous woody shrubs, O. europaea and G. umbellata were the slowest in decomposing (1.18 and 1.01 years⁻¹, respectively) contrary to L. officinalis, which decomposed fastest (2.22 years⁻¹). Only L. officinalis and P. purpurea registered a net N-release at the end of the study. The AMMPs showed different decomposition patterns: L. dentata registered the highest decomposition rates and Rosmarinus the lowest (1.9 and 1.1 years⁻¹, respectively). T. mastichina, L. dentata, and S. obovata had the highest N-release, whereas R. officinalis and A. cytisoides showed N immobilization (183 and 122% of the initial N). Knowledge of the dynamics of nutrient release and litter decomposition will be useful for predicting nutrient availability and nutrient cycles in these types of agroecosystems where subtropical orchards are grown on terraces.     

Recilia banda Kramer (Hemiptera: Cicadellidae), a vector of Napier stunt phytoplasma in Kenya

Napier grass (Pennisetum purpureum) is the most important fodder crop in smallholder dairy production systems in East Africa, characterized by small zero-grazing units. It is also an important trap crop used in the management of cereal stemborers in maize in the region. However, production of Napier grass in the region is severely constrained by Napier stunt disease. The etiology of the disease is known to be a phytoplasma, 16SrXI strain. However, the putative insect vector was yet unknown. We sampled and identified five leafhopper and three planthopper species associated with Napier grass and used them as candidates in pathogen transmission experiments. Polymerase chain reaction (PCR), based on the highly conserved 16S gene, primed by P1/P6-R16F2n/R16R2 nested primer sets was used to diagnose phytoplasma on test plants and insects, before and after transmission experiments. Healthy plants were exposed for 60 days to insects that had fed on diseased plants and acquired phytoplasma. The plants were then incubated for another 30 days. Nested PCR analyses showed that 58.3% of plants exposed to Recilia banda Kramer (Hemiptera: Cicadellidae) were positive for phytoplasma and developed characteristic stunt disease symptoms while 60% of R. banda insect samples were similarly phytoplasma positive. We compared the nucleotide sequences of the phytoplasma isolated from R. banda, Napier grass on which these insects were fed, and Napier grass infected by R. banda, and found them to be virtually identical. The results confirm that R. banda transmits Napier stunt phytoplasma in western Kenya, and may be the key vector of Napier stunt disease in this region.     

Soil microbial response to herbicides applied to glyphosate-resistant canola

Adoption of glyphosate-resistant canola (Brassica napus L.) has increased glyphosate applications to this crop, and concerns have been raised about unintended consequences of these multiple applications. A field trial was conducted to evaluate the effects of pre-seed and in-crop glyphosate and alternative herbicides on soil microbial community functional structure, diversity and biomass. Pre-seed treatments were 2,4-D, glyphosate and 2,4-D + glyphosate, and in-crop treatments were glyphosate applied once, glyphosate applied twice, ethalfluralin, ethalfluralin + sethoxydim + ethametsulfuron + clopyralid, and sethoxydim + ethametsulfuron. Rhizosphere and bulk soil was collected at flowering stage of canola and analyzed for bacterial community-level substrate utilization patterns and microbial biomass C (MBC). Where differences were significant, pre-seed application of both 2,4-D and glyphosate altered the functional structure and reduced the functional diversity of soil bacteria, but increased MBC. These effects were not necessarily concurrent. The reduction in functional diversity was due to reduction in evenness, which means that the soil where both pre-seed herbicides had been applied was dominated by only few functional groups. In 1 year, two in-crop applications of glyphosate also reduced the functional diversity of soil bacteria when applied after pre-seed 2,4-D, as did in-crop sethoxydim + ethametsulfuron following pre-seed glyphosate. Even though significant differences between herbicides were fewer than non-significant differences, i.e., there were no changes in soil microbial community structure, diversity or biomass in response to glyphosate or alternative herbicides applied to glyphosate-resistant canola in most cases, the observed changes in soil microbial communities could affect soil food webs and biological processes.     

The antennal sensilla of Melipona quadrifasciata (Hymenoptera: Apidae: Meliponini): a study of different sexes and castes

The sensilla of insects are integumental units that play a role as sensory structures and are crucial for the perception of stimuli and for communication. In this study, we compared the antennal sensilla of females (workers and queens), males (haploid (n) and diploid (2n)), and queen-like males (QLMs, resulting from 2n males after juvenile hormone (JH) treatment) in the stingless bee Melipona quadrifasciata. Images of the dorsal antenna surfaces were acquired using a scanning electron microscope. As reported for other hymenopterans, this species exhibits a heterogeneous sensillar distribution along the antennae. Thirteen different types of sensilla were found in the antennae of M. quadrifasciata: trichodea (subtypes I to VI), chaetica (subtypes I and II), placodea, basiconica, ampullacea, coeloconica, and coelocapitula. Sensilla trichodea I were the most abundant, followed by sensilla placodea, which might function in olfactory perception. Sensilla basiconica, sensilla chaetica I, sensilla coeloconica, and sensilla ampullacea were found exclusively in females. In terms of the composition and size of the sensilla, the antennae of QLMs most closely resemble those of the 2n male, although QLMs exhibit a queen phenotype. This study represents the first comparative analysis of the antennal sensilla of M. quadrifasciata. The differences found in the type and amount of sensilla between the castes and sexes are discussed based on the presumed sensillary functions.     

Qualitative and Quantitative Assessment of Hepatitis A Virus in Wastewaters in Tunisia

Hepatitis A causes substantial morbidity in both industrialized and non-industrialized countries and represents an important health problem in several southern Mediterranean countries. The objectives of the study were as follows: (a) to assess the occurrence of hepatitis A virus (HAV) in Tunisia through the monitoring of urban wastewaters collected at wastewater treatment plants (WTPs); (b) to characterize environmental strains; and (c) to estimate the viral load in raw and treated sewages, in order to evaluate the potential impact on superficial waters receiving discharges. A total of 150 raw and treated wastewaters were collected from three WTPs and analyzed by both qualitative (RT-PCR/nested) and quantitative (qRT-PCR) methods. Of these, 100 (66 %) were found to be positive for HAV by the qualitative assay: 68.3 % in influents and 64.7 % in effluents. The vast majority of HAV sequences belonged to sub-genotype IA, with 11 different strains detected found to be identical to clinical strains isolated from Tunisian patients with acute hepatitis. Five unique variants were also detected, not previously reported in clinical cases. Only two IB strains were found, confirming the rarity of this sub-genotype in this country. The results of the present study indicate a wide circulation of the pathogen in the population, most probably in the form of asymptomatic infections, a finding consistent with the classification of the country as having intermediate/high endemicity. Quantitative data showed high viral loads in influents (3.5E+05 genome copies/liter, mean value) as well as effluents (2.5E+05 genome copies/liter, mean value), suggesting that contaminated water could be a critical element in transmission.     

Leaf and fruit necroses associated with vanadium-rich ash emitted from a power plant burning fossil fuel

Necrotic brown lesions developing on leaves and fruits of lemon, and other plants, near fossil-fuel-burning power plants emitting vanadium-rich ash, were reproduced experimentally by inoculating leaves with this type of ash, which is strongly acidic. Comparable lesions were formed when drops of vanadium oxide, dissolved in NaOH, were put on leaves and fruits. Neutralizing, or adding excess H₂SO₄ (to pH 3), did not alter lesion formation. The histology of these lesions was similar to that of lesions attributed to vanadium-rich ash.     

Humic fractions of organic wastes

The distribution and composition of humic and fulvic acids have been investigated in the following organic wastes: straw, farmyard manure, sewage sludge, poultry manure, municipal refuse compost and pig slurry. Of the total organic carbon, the proportion present as humic carbon varied from about 13% in the compost to 50% in the pig slurry; the humic compounds extracted are characterized by a variable chemical composition. As compared to the natural humic substances from soil organic matter, these compounds show a lower content of acidic functional groups, lower E₄/E₆ and C/H ratios and can be classified as young forms of humic substances.