Herbicide risk assessment for non-target aquatic plants: sulfosulfuron--a case study

Herbicides entering the aquatic environment by spray drift, run-off and leaching to field drains may cause adverse effects on non-target aquatic vegetation. The potential for such effects has typically been evaluated from tests with floating, monocotyledonous Lemna sp. However, concern has been expressed as to whether this species could be used to indicate potential effects on other vegetation types, particularly rooted, submerged, emergent or dicotyledonous species. In 1997, the Centre for Aquatic Plant Management undertook development of new tests based on the additional species, Glyceria maxima (Hartm) Holmb, Lagarosiphon major (Ridl) Moss and Myriophyllum spicatum L. The resulting methodology was used to assess the effects of the sulfonylurea herbicide, sulfosulfuron on these species. Data presented here demonstrate that exposure to initial sulfosulfuron concentrations of 3.33 microg litre(-1) for up to 21 days was tolerated by these species and that adverse effects were observed only when plants were exposed to initial concentrations of 3.33 and 10 microg litre(-1) for 70 days. As the occurrence of such high initial concentrations for long periods is unlikely in the aquatic environment, sulfosulfuron is not expected to have adverse effects on the growth of these species. This study has also demonstrated that G maxima, L major and M spicatum grown in small outdoor tanks can be used successfully to assess the effects of crop-protection products on non-target aquatic flora.     

Evaluation of biological quality of protein concentrates from different vegetable protein sources

The quality of protein concentrates coming from conventional and non-conventional vegetable sources was evaluated. The protein quality was assessed by the multi-point assay RPV. Whole body weight gain, weight gain corrected for the gastrointestinal content (G.I.), total body nitrogen and total liver nitrogen were used as response parameters. The various protein sources ranked differently depending upon the response parameters utilized, but a great majority of the sources had a good nutritional value (over 0.5). The gastrointestinal content is one of the principal factors responsible for the differences found. However, even after correcting for the G.I. content, some differences remained. For the various protein sources, the nitrogen deposition in the liver correlates satisfactorily with the whole body nitrogen deposition, if in the calculation of the regression line, the data for faba bean and tobacco were omitted (respectivelyr=0.610 andr=0.925). On the whole, these data indicate that most plant protein concentrates in suitable combinations and properly processed, may become a promising alternative to animal proteins.     

Co-operational PCR coupled with dot blot hybridization for detection and 16SrX grouping of phytoplasmas

A protocol based on Co-operational PCR has been successfully applied to the detection of phytoplasmas. A triprimer reaction coupled with hybridization using general and specific probes permitted detection of ' Candidatus Phytoplasma mali', ' Ca . Phytoplasma prunorum' and ' Ca . Phytoplasma pyri', and their identification as members of 16S ribosomal quarantine group X. The sensitivity of this method was at least one hundred times greater than conventional PCR and similar to that achieved by nested PCR and real-time PCR. The method was validated by testing field samples collected from Malus , Prunus and Pyrus spp. and Olea europaea and compared with seven phytoplasmas maintained in Catharanthus roseus .     

Factors Affecting Pseudomonas savastanoi pv. savastanoi Plant Inoculations and Their Use for Evaluation of Olive Cultivar Susceptibility

ABSTRACT Pseudomonas savastanoi pv. savastanoi causes olive knot disease, which is present in most countries where olive trees are grown. Although the use of cultivars with low susceptibility may be one of the most appropriate methods of disease control, little information is available from inoculation assays, and cultivar susceptibility assessments have been limited to few cultivars. We have evaluated the effects of pathogen virulence, plant age, the dose/response relationship, and the induction of secondary tumors in olive inoculation assays. Most P. savastanoi pv. savastanoi strains evaluated were highly virulent to olive plants, but interactions between cultivars and strains were found. The severity of the disease in a given cultivar was strongly dependent of the pathogen dose applied at the wound sites. Secondary tumors developed in noninoculated wounds following inoculation at another position on the stem, suggesting the migration of the pathogen within olive plants. Proportion and weight of primary knots and the presence of secondary knots were evaluated in 29 olive cultivars inoculated with two pathogen strains at two inoculum doses, allowing us to rate most of the cultivars as having either high, medium, or low susceptibility to olive knot disease. None of the cultivars were immune to the disease.     

Harmonised pesticide risk trend indicator for food (HAPERITIF): The methodological approach

To provide a harmonised European approach for pesticide risk indicators, the Sixth EU Framework Programme recently financed the HAIR (HArmonised environmental Indicators for pesticide Risk) project. This paper illustrates the methodology underlying a new indicator-HAPERITIF (HArmonised PEsticide RIsk Trend Indicator for Food), developed in HAIR, for tracking acute and chronic pesticide risk trends for consumers. The acute indicator, HAPERITIF(ac), is based on the ratio between an estimated short-term intake (ESTI), calculated as recommended by the World Health Organisation (WHO), and the acute reference dose (ARfD); the chronic indicator HAPERITIF(chr) is based on the ratio between an estimated daily intake (EDI) and the admissible daily intake (ADI). HAPERITIF can be applied at different levels of aggregation. Each level gives information for proper risk management of pesticides to reduce the risk associated with food consumption. An example of application using realistic scenarios of pesticide treatments on a potato crop in central-northern Italy is reported to illustrate the different steps of HAPERITIF.     

A genome-wide analysis of cardiac lesions of pigs that die during transport: Is heart failure of in-transit-loss pigs associated with a heritable cardiomyopathy?

While heart failure is a primary cause of death for many in-transit-loss (ITL) pigs, the underlying cause of these deaths is not known. Cardiomyopathies are considered a common cause of heart failure in humans and often have a genetic component. The objective of this study was to determine if genes associated with cardiomyopathies could be identified in ITL pigs. Samples from the hearts of pigs that died during transport to an abattoir in Ontario, Canada were collected and genotyped along with samples from pigs that did not die during transport (ILT hearts: n = 149; non-ITL/control hearts: n = 387). Genome-wide analyses were carried out on each of the determined phenotypes (gross cardiac lesions) using a medium density single nucleotide polymorphism (SNP) chip and 500 kb windows/regions for analysis, with 250 kb regions of overlap. The distribution derived by a multidimensional scaling (MDS) analysis of all phenotypes demonstrated a lack of complete separation between phenotypes of affected and unaffected animals, which made diagnosis difficult. Although genetic differences were small, a few genes associated with dilated cardiomyopathy (DCM) and arrhythmogenic right ventricular cardiomyopathy (ARVM) were identified. In addition, multiple genes associated with cardiac arrhythmias and ventricular hypertrophy were identified that can possibly result in heart failure. The results of this preliminary study did not provide convincing evidence that a single, heritable cardiomyopathy is the cause of heart failure in ITL pigs.     

Proteomic profile of pre-implantational ovine embryos produced in vivo

The present study was conducted to decipher the proteome of in vivo-produced pre-implantation ovine embryos. Ten locally adapted Morana Nova ewes received hormonal treatment and were inseminated 12 hr after ovulation. Six days later, 54 embryos (morula and blastocyst developmental state) were recovered from eight ewes and pooled to obtain sufficient protein for proteomic analysis. Extracted embryo proteins were analysed by LC-MS/MS, followed by identification based on four database searches (PEAKS, Proteome Discoverer software, SearchGUI software, PepExplorer). Identified proteins were analysed for gene ontology terms, protein clusters and interactions. Genes associated with the ovine embryo proteome were screened for miRNA targets using data sets of TargetScan ( http://www.targetscan.org ) and mIRBase ( http://www.mirbase.org ) servers. There were 667 proteins identified in the ovine embryos. Biological processes of such proteins were mainly related to cellular process and regulation, and molecular functions, to binding and catalytic activity. Analysis of the embryo proteins revealed 49 enriched functional clusters, linked to energy metabolism (TCA cycle, pyruvate and glycolysis metabolism), zona pellucida (ZP), MAPK signalling pathway, tight junction, binding of sperm to ZP, translation, proteasome, cell cycle and calcium/phospholipid binding. Sixteen miRNAs were related to 25 pre-implantation ovine embryo genes, all conserved in human, bovine and ovine species. The interaction network generated by miRNet showed four key miRNAs (hsa-mir-106b-5p; hsa-mir-30-5p; hsa-mir-103a-5p and hsa-mir-106a-5p) with potential interactions with embryo-expressed genes. Functional analysis of the network indicated that miRNAs modulate genes related to cell cycle, regulation of stem cell and embryonic cell differentiation, among others. Retrieved miRNAs also modulate the expression of genes involved in cell signalling pathways, such as MAPK, Wnt, TGF-beta, p53 and Toll-like receptor. The current study describes the first major proteomic profile of 6-day-old ovine embryos produced in vivo, setting a comprehensive foundation for our understanding of embryo physiology in the ovine species.     

Analysis of fatty acid steryl esters in tetraploid and hexaploid wheats: identification and comparison between chromatographic methods

Fatty acid steryl esters (FASE) in whole meal of 14 genotypes of tetraploid wheats (Triticum dicocconand T. durum) and 17 genotypes of hexaploid wheats (T. spelta and T. aestivum) were analyzed using different chromatographic strategies. By both GC-FID and HPLC-ELSD, tetraploid wheats are lacking two major peaks. The amounts of FASE, calculated on the basis of the GC-FID analysis, were double in hexaploid species as compared to tetraploids (40 and 20 mg/100 g db, respectively). HPLC with ESI-MS detection enabled the identification of FASE by the characteristic fragmentations and ion-adducts of each molecule. The distribution of steryl residues was not different between the wheat species: the main class of steryl derivatives found was the beta-sitosteryl derivatives, followed by campesteryl derivatives with small amounts of stigmasteryl esters. The esterified fatty acids explain the difference between the hexaploid and tetraploid wheats. In particular, small amounts of campesteryl and beta-sitosteryl, while no trace of stigmasteryl palmitates, were found in T. durum or its hulled ancestor T. dicoccon. Steryl oleates were not detectable in T. aestivum or its hulled ancestor T. spelta, which is consistent with the filogenesis of tetraploid and hexaploid species. Both chromatographic techniques (GC and HPLC) showed that FASE are useful to discriminate between hexaploid and tetraploid wheats from both qualitative and quantitative points of view.     

Effects of glucosinolates and their enzymatic hydrolysis products via myrosinase on the root-knot nematode Meloidogyne incognita (Kofoid et White) Chitw

The root-knot nematode Meloidogyne incognita (Kofoid et White) Chitw. is responsible for large yield losses in several horticultural crops. Fumigation with chemicals has been efficient in fighting this soil pest, but it clearly shows a negative environmental impact. Thus, it is necessary to find an environmentally friendly alternative to control this nematode and meet the requirements imposed by world regulation to ban some chemical fumigants in the world after 2005. The glucosinolate-myrosinase system, typical of the Brassicaceae family, appears to be an important natural alternative for the control of several soilborne pests and pathogens. The aim of this study was to evaluate, in vitro, the biocidal activity of 11 glucosinolates and their degradation products on second-stage juveniles of the root-knot nematode M. incognita expressed by the nematicidal (LD(50)) and immobilization effects, after 24 and 48 h. None of the intact glucosinolates had any biological effect. After myrosinase addition, their hydrolysis products (essentially isothiocyanates) resulted in highly different biocidal activities. Among the hydrolysis products of the tested glucosinolates, 2-phenylethyl, benzyl, 4-methylthiobutyl, and prop-2-enyl isothiocyanate showed the stronger activity, with an LD(50) at concentrations of 11, 15, 21, and 34 microM, respectively. On the basis of the in vitro test results, new genotypes of Brassicaceae had been selected for high content in the roots of the glucosinolates generating the more active isothiocyanates and their agronomic performances verified in view of a full-field application as catch crop plants. With this aim, the qualitative and quantitative glucosinolate contents in the roots of these potentially nematicidal plants are also reported and discussed.