Efficacy of metalaxyl for blue mould control and its persistence on tobacco

Foliar sprays of metalaxyl, benalaxyl, and cymoxanil plus mancozeb gave better control of blue mould (Peronospora tabacina) than mancozeb alone applied as a spray, or metalaxyl applied to the soil. Before flue-curing, the mean values of metalaxyl residues in tobacco leaves were significantly higher from foliar spray treatments (5.09 μg g⁻¹), than from soil treatments (0.93 μg g⁻¹). Residues had decreased after flue-curing (foliar spray treatment 2.51 μg g⁻¹; soil treatment 0.69 μg g⁻¹), especially on samples taken in September. Curing considerably reduced metalaxyl residues in all cases. Residues of mancozeb ranged from 59.5 to 224.2 μg g⁻¹ before flue-curing and from 11.0 to 22.1 μg g⁻¹ after flue-curing. All these residues were calculated on a dry weight basis. Imidazolidine-2-thione (ethylenethiourea) residues from mancozeb were always below the sensitivity limit of the method used.     

Characterization and genetic analysis of field isolates ofBotryotinia fuckeliana (Botrytis cinerea) resistant to dichlofluanid

Field isolates ofBotryotinia fuckeliana were collected from naturally infected plants. Their responses to the multisite fungicide dichlofluanid in mycelium growth test fell into three phenotypic classes, characterized by the following EC₅₀ (and MIC) values ing ml^–1: sensitivity, 1–3 (6–10); low resistance, 3–10 (> 100); high resistance, 10–30 (> 100). The corresponding values obtained for these classes in a spore germination test were respectively: 0.05 (0.2), 0.05–0.1 (0.5), 0.5–1 (0.9–1.5). Resistant isolates were crossed with two sensitive and two resistant strains of appropriate mating type to determine the genetic basis of resistance. Distribution of resistance phenotypes in ascospore progeny indicated that a gene, namedDic1, was probably responsible for the low or high resistance of 14 mutants selectively collected from experimental plots of greenhouse-grown gerbera sprayed several times with dichlofluanid or tolyfluanid. A second gene, namedDic2, was probably responsible for the low resistance displayed by two isolates (from grapevine and from carnation) maintained in the laboratory collection. As a result of the investigation, the use of dichlofluanid in integrated management programmes against grey mould is discussed.     

Combined interventional procedure and cardiopulmonary bypass surgery in a dog with cor triatriatum dexter,patent foramen ovale,and pulmonary stenosis

A 2‐year‐old American Pit Bull dog was presented for surgical evaluation of imperforate cor triatriatum dexter (CTD) and patent foramen ovale (PFO). Echocardiography identified an imperforate CTD associated with a right‐to‐left shunting PFO and valvular pulmonary stenosis. A 2‐step interventional and surgical approach was used. Initially, a pulmonary balloon valvuloplasty was performed, and subsequently the dog underwent a surgical correction of the atrial anomaly under cardiopulmonary bypass.     

The role of parameterization in comparing source attribution models based on microbial subtyping for salmonellosis

Source attribution methods attribute human cases of a zoonotic disease to a certain source putatively responsible for this disease. Identifying and quantifying the contribution of different sources to human infections is important for taking appropriate actions for reducing the exposure of the consumer to zoonotic pathogens. One widely used method is the microbial subtyping approach, whose principle is to compare the frequency of pathogen subtypes from different sources (e.g. animals or food) with the frequency of these subtypes in human cases. This paper studies the relationship between a Bayesian microbial subtyping approach described by Hald and coworkers subsequently modified by David and coworkers, here called the Hald model, and a frequentist approach known as the “Dutch model.” The comparison between the Bayesian and frequentist model is done for two data sets on salmonellosis in Germany from different time periods (year 2004–2007 and 2010–2011). The results of both approaches are in good agreement with each other for the used data. It is shown here mathematically that a certain parameterization can be used to transform the probabilistic Hald model into a deterministic form, which is equivalent to the Dutch model. That certain parameterization secures independence of the model outcomes from the choice of so‐called unique subtypes (which are unique in the sense that they are found exclusively in one of the sources). It is shown that deviating from that certain parameterization leads variations in the model outcome dependent on which unique subtypes are chosen in the process of modelling.     

Integrated standardization concept for Angelica botanicals using quantitative NMR

Despite numerous in vitro/vivo and phytochemical studies, the active constituents of Angelica sinensis (AS) have not been conclusively identified for the standardization to bioactive markers. Phytochemical analyses of AS extracts and fractions that demonstrate activity in a panel of in vitro bioassays, have repeatedly pointed to ligustilide as being (associated with) the active principle(s). Due to the chemical instability of ligustilide and related issues in GC/LC analyses, new methods capable of quantifying ligustilide in mixtures that do not rely on an identical reference standard are in high demand. This study demonstrates how NMR can satisfy the requirement for simultaneous, multi-target quantification and qualitative identification. First, the AS activity was concentrated into a single fraction by RP-solid-phase extraction, as confirmed by an alkaline phosphatase, (anti-)estrogenicity and cytotoxicity assay. Next, a quantitative ¹H NMR (qHNMR) method was established and validated using standard compounds and comparing processing methods. Subsequent 1D/2D NMR and qHNMR analysis led to the identification and quantification of ligustilide and other minor components in the active fraction, and to the development of quality criteria for authentic AS preparations. The absolute and relative quantities of ligustilide, six minor alkyl phthalides, and groups of phenylpropanoids, polyynes, and poly-unsaturated fatty acids were measured by a combination of qHNMR and 2D COSY. The qNMR approach enables multi-target quality control of the bioactive fraction, and enables the integrated biological and chemical standardization of AS botanicals. This methodology can potentially be transferred to other botanicals with active principles that act synergistically, or that contain closely related and/or constituents, which have not been conclusively identified as the active principles.     

Influence of storage approaches on instability of propiconazole resistance in Monilinia fructicola

BACKGROUND: The long‐term preservation of interesting phenotypes in plant pathogenic fungi allows for follow‐up studies in the future. Twelve storage approaches were investigated to determine their effects on instability of propiconazole resistance for three demethylation inhibitor (DMI) fungicide‐resistant and two DMI‐sensitive isolates of Monilinia fructicola. They included mycelium in PDA slants under mineral oil, in PDA plugs under 10% glycerol, on dried filter paper and conidia on silica gel, each stored for 36 weeks at 4, ? 20, and ? 80 °C. RESULTS: None of the storage approaches prevented the rapid decline of EC₅₀ values for propiconazole in the three resistant isolates, and no significant differences were found among storage approaches (P = 0.787) or between storage approaches and consecutive transfers (P = 0.053). Most of the decline in resistance occurred during the first 4 weeks of storage. The DMI resistance‐associated genetic element Mona, located in the immediate upstream region of the MfCYP51 gene, was still present in the three resistant isolates after 36 weeks of storage and weekly transfers. Furthermore, the Mona element and a portion of the MfCYP51 gene, which encodes the target enzyme for DMIs, did not reveal signs of DNA methylation. Resistance to propiconazole was partially regained in resistant isolates after two growth cycles on fresh peach fruit. CONCLUSIONS: Obtained data indicate that the decline of DMI resistance in M. fructicola cannot be prevented using commonly employed storage methods at various temperatures. The number of consecutive transfers and the storage duration prior to fungicide sensitivity tests in M. fructicola should be indicated in scientific papers. Copyright © 2011 Society of Chemical Industry     

Benefits and well-being perceived by people visiting green spaces in periods of heat stress

In urban environments, green spaces have proven to act as ameliorating factors of some climatic features related to heat stress, reducing their effects and providing comfortable outdoor settings for people. In addition, green spaces have demonstrated greater capacity, compared with built-up areas, for promoting human health and well-being. In this paper, we present results of a study conducted in Italy and the UK with the general goal to contribute to the theoretical and empirical rationale for linking green spaces with well-being in urban environments. Specifically, the study focused on the physical and psychological benefits and the general well-being associated with the use of green spaces on people when heat stress episodes are more likely to occur. A questionnaire was set up and administered to users of selected green spaces in Italy and the UK (n=800). Results indicate that longer and frequent visits of green spaces generate significant improvements of the perceived benefits and well-being among users. These results are consistent with the idea that the use of green spaces could alleviate the perception of thermal discomfort during periods of heat stress.     

Prevalence of Ehrlichia canis infection in thrombocytopenic dogs from Rio de Janeiro, Brazil

BACKGROUND: Infection with Ehrlichia canis causes a highly variable, multisystemic disease in dogs. Nevertheless, many clinicians in Rio de Janeiro, Brazil, use the presence of only thrombocytopenia to make a presumptive diagnosis of E canis infection. OBJECTIVE: The objective of this study was to determine the prevalence of E canis in thrombocytopenic dogs from Rio de Janeiro, Brazil, using polymerase chain reaction (PCR). METHODS: Following DNA extraction of whole blood samples from 226 dogs, PCR assays were done using primers for rickettsial DNA (including Ehrlichia spp, Anaplasma platys and A phagocytophilum) and using E canis-specific primers (16S rRNA gene). Dogs were grouped as thrombocytopenic and nonthrombocytopenic based on platelet counts. The null hypothesis that there was no difference in the prevalence of E canis in these groups was rejected at P<.05. RESULTS: Thirty-six (32.1%) of the thrombocytopenic dogs and 4 (3.5%) of the nonthrombocytopenic dogs were positive for rickettsial gene sequences (P<.0001). Further, 30 (26.8%) of thrombocytopenic dogs and 4 (3.5%) nonthrombocytopenic dogs were positive for E canis-specific gene sequences (P<.0001). CONCLUSIONS: Although the prevalence of E canis infection was higher in thrombocytopenic dogs, less than one third of these dogs had demonstrable E canis infection. Thus, thrombocytopenia is not specific for the detection of E canis infection and should not be used solely to establish a diagnosis of canine ehrlichiosis, even in a geographic area with relatively high disease prevalence.     

Biochemical characterisation of some non fermenting, non arginine hydrolysing mycoplasmas of ruminants

The pattern and kinetics of substrate oxidation by type and recent field strains of Mycoplasma agalactiae, Mycoplasma bovis, Mycoplasma bovigenitalium and Mycoplasma ovine/caprine serogroup 11 were investigated by measurement of oxygen uptake. Metabolism of a range of organic acids, sugars and alcohols was detected. All the test strains were unable to oxidise sugars, glycerol and the organic acids, fumarate, malate and alpha-ketoglutarate (1 mM). All strains oxidised organic acid l-lactate, 2-oxobutyrate and pyruvate and demonstrated the ability to oxidise alcohols, particularly isopropanol, which was oxidised at a high rate and high affinity (0.5 mol/mol isopropanol). Its oxidation was consistent with acetone formation, which may be of important in relation to pathogenicity. All strains oxidised similar substrates, however differences were observed between strains in terms of the relative rates and kinetic values for some substrates.