Visualization and functional analysis of RNA-dependent RNA polymerase lattices

Positive-strand RNA viruses such as poliovirus replicate their genomes on intracellular membranes of their eukaryotic hosts. Electron microscopy has revealed that purified poliovirus RNA-dependent RNA polymerase forms planar and tubular oligomeric arrays. The structural integrity of these arrays correlates with cooperative RNA binding and RNA elongation and is sensitive to mutations that disrupt intermolecular contacts predicted by the polymerase structure. Membranous vesicles isolated from poliovirus-infected cells contain structures consistent with the presence of two-dimensional polymerase arrays on their surfaces during infection. Therefore, host cytoplasmic membranes may function as physical foundations for two-dimensional polymerase arrays, conferring the advantages of surface catalysis to viral RNA replication.     

Health of farmed fish: its relation to fish welfare and its utility as welfare indicator

This brief review focuses on health and biological function as cornerstones of fish welfare. From the function-based point of view, good welfare is reflected in the ability of the animal to cope with infectious and non-infectious stressors, thereby maintaining homeostasis and good health, whereas stressful husbandry conditions and protracted suffering will lead to the loss of the coping ability and, thus, to impaired health. In the first part of the review, the physiological processes through which stressful husbandry conditions modulate health of farmed fish are examined. If fish are subjected to unfavourable husbandry conditions, the resulting disruption of internal homeostasis necessitates energy-demanding physiological adjustments (allostasis/acclimation). The ensuing energy drain leads to trade-offs with other energy-demanding processes such as the functioning of the primary epithelial barriers (gut, skin, gills) and the immune system. Understanding of the relation between husbandry conditions, allostatic responses and fish health provides the basis for the second theme developed in this review, the potential use of biological function and health parameters as operational welfare indicators (OWIs). Advantages of function- and health-related parameters are that they are relatively straightforward to recognize and to measure and are routinely monitored in most aquaculture units, thereby providing feasible tools to assess fish welfare under practical farming conditions. As the efforts to improve fish welfare and environmental sustainability lead to increasingly diverse solutions, in particular integrated production, it is imperative that we have objective OWIs to compare with other production forms, such as high-density aquaculture. However, to receive the necessary acceptance for legislation, more robust scientific backing of the health- and function-related OWIs is urgently needed.     

Top-spraying soybean meal-based diets with phytase improves protein and mineral digestibilities but not lysine utilization in rainbow trout, Oncorhynchus mykiss (Walbaum)

Two digestibility trials and two growth trials were carried out to evaluate the influence of top‐sprayed phytase on apparent digestibility coefficients (ADCs) of protein and mineral and utilization in rainbow trout fed with soybean meal‐based diets. In Trial 1, a semi‐purified diet containing 50% soybean meal was supplemented with graded levels of phytase (0, 500, 1000, 2000 and 4000 U kg^?1 diet), and fed to triplicate groups of fish. In Trial 2, commercial‐type extruded feeds containing 36% soybean meal with either 0 or 2000 U phytase kg^?1 were fed to five replicate groups of fish. Phytase clearly decreased phytic acid content of feces from 35 to 5 mg and from 34 to 14 mg phytic acid per g faecal dry matter in Trials 1 and 2 respectively. Apparent digestibility coefficient of P improved from 23% to 83% in Trial 1 and from 35% to 54% in Trial 2 by phytase. Apparent protein increased by 1.2% and 3.2%‐units by phytase in Trials 1 and 2. Zinc digestibility was significantly increased in Trial 1, but not in Trial 2. Trials 3 and 4 were conducted to evaluate the influence of phytase on dietary P (Trial 3) and lysine (Trial 4) utilization. Three diets were prepared for each trial: P (Trial 3)‐ or lysine (Trial 4)‐deficient basal diets, basal diets with phytase supplementation (2000 U kg^?1) and P (Trial 3)‐ or lysine (Trial 4)‐fortified diets. Rainbow trout (initial weight 20 g) were fed for 10 weeks using four and six replicates for Trials 3 and 4 respectively. Phytase increased P utilization in Trial 3 as demonstrated by an increase in vertebra ash from 24.1% to 45.4%, and by an increase in weight gain from 243% to 459% of the initial weight. Phytase did not increase lysine utilization, since neither protein retention nor weight gain were enhanced by phytase. Supplemental lysine increased protein retention and weight gain to 43.1% and 514%, respectively, and also decreased whole‐body lipid contents significantly from 120 to 123 g kg^?1 in fish fed the basal diet and phytase‐supplemented diet to 106 g kg^?1 in fish fed with lysine‐fortified diet.     

Fluoreszenzmikroskopischer Nachweis von DDT und seinen Mitteln

Ohne Zusammenfassung     

Breeding objectives for indigenous chicken: Model development and application to different production systems

A bio-economic model was developed to evaluate the utilisation of indigenous chickens (IC) under different production systems accounting for the risk attitude of the farmers. The model classified the production systems into three categories based on the level of management: free-range system (FRS), where chickens were left to scavenge for feed resources with no supplementation and healthcare; intensive system (IS), where the chickens were permanently confined and supplied with rationed feed and healthcare; and semi-intensive system (SIS), a hybrid of FRS and IS, where the chickens were partially confined, supplemented with rationed feeds, provided with healthcare and allowed to scavenge within the homestead or in runs. The model allows prediction of the live weights and feed intake at different stages in the life cycle of the IC and can compute the profitability of each production system using both traditional and risk-rated profit models. The input parameters used in the model represent a typical IC production system in developing countries but are flexible and therefore can be modified to suit specific situations and simulate profitability and costs of other poultry species production systems. The model has the capability to derive the economic values as changes in the genetic merit of the biological parameter results in marginal changes in profitability and costs of the production systems. The results suggested that utilisation of IC in their current genetic merit and production environment is more profitable under FRS and SIS but not economically viable under IS.     

Pathogenicity,Morpho-Species and Mating Types of <Emphasis Type="Italic">Alternaria</Emphasis> spp. causing Alternaria blight in <Emphasis Type="Italic">Pistacia</Emphasis> spp. in Turkey

Alternaria genus includes many plant pathogens on numerous hosts, causing leaf spots, rots and blights. Alternaria blight has been observed as one of the important fungal diseases of pistachio (Pistacia vera L.) as well as its wild relatives (P. terebinthus, P. lentiscus, P. khinjuk, P. atlantica, P. mutica) in Turkey. Alternaria species were sampled from Pistacia spp. hosts from different geographic regions in Turkey during field trips in late spring to early fall of 2013. Alternaria blight symptoms were observed mainly on fruits and rarely on leaves. Four hundred and twenty two of the isolates were morphologically defined as A. alternata, A. tenuissima, A. arborescens and also intermediate morpho-species between A. alternata/A. arborescens. Pathogenicity of the isolates was confirmed with host inoculations on detached fruits. Mating types of 270 isolates of Alternaria spp. from the collection were identified using a PCR-based mating type assay that amplifies either a MAT1-1 or a MAT1-2 fragment from the mating locus. Although a strongly clonal population structure was expected due to the putative asexual reproduction of these fungi, both idiomorphs were detected at equal frequencies at several different spatial scales. The distribution of mating types within each geographic region, within host species as well as in overall collection was not significantly different from 1:1. Amplified fragments of partial idiomorph sequences were obtained for representative isolates. Parsimony trees were depicted based on sequence data of mating type genes for these representative isolates as well as some other Alternaria species obtained by Genebank. Several point mutations presented a few clusters which are supported by high bootsrapped values. The Alternaria blight disease agents both from cultivated and wild hosts were pathogenic on pistachio which may cause difficulties to control the disease because of extensity of pathogen sources. Besides, equal mating type distribution of the pathogen at both geographic and host species levels suggests a potential for sexual reproduction of Alternaria spp. in Turkey.     

In vitro and in vivo antifungal activity of synthetic pure isothiocyanates against Sclerotinia sclerotiorum

BACKGROUND: Isothiocyanates (ITCs) released by the enzymatic hydrolysis of glucosinolates in the Brassicaceae are potentially useful for controlling fungal pathogens. In vitro activity of pure ITCs against Sclerotinia sclerotiorum (Lib.) de Bary was studied by adding them to glass filters in petri dishes and dissolving them in the growing media. RESULTS: Methyl, allyl and benzyl ITCs were the most fungitoxic of the compounds in bioassays with S. sclerotiorum isolate Ss31. In the volatile phase, mycelial growth was completely inhibited by these three compounds. Aromatic ITCs were less toxic in the petri dishes but were more toxic than aliphatic ITCs when dissolved in the agar. Benzyl ITC exhibited the highest inhibitory effect on sclerotial germination, with an EC₅₀ value of 75.1 µmol L^?1. Butyl and benzyl ITCs reduced apothecial production of S. sclerotiorum by 92.5% at the highest concentration. In in vivo assay, only allyl and 2‐phenylethyl ITCs reduced disease incidence (by 76.7 and 70% respectively) at low concentrations. CONCLUSION: Sclerotinia sclerotiorum in the soil might be suppressed by the higher concentrations of allyl and benzyl ITCs released from decomposition of Brassica juncea, B. carinata, B. nigra and Sinapis spp. Copyright © 2011 Society of Chemical Industry