Estimation of Genetic Variation Among Verticillium Isolates using AFLP Analysis

Amplified fragment length polymorphism (AFLP) analysis was used to study genetic variation among 76 isolates of Verticillium. A dendrogram based on the AFLP data revealed three main groups. One group consisted of 35 European isolates derived from Brassica napus together with five Californian isolates taken from B. oleracea. This group displayed a high degree of genetic similarity and included three isolates earlier classified as Verticillium longisporum, indicating that all isolates in this group probably should be regarded as members of V. longisporum. V. dahliae isolates constituted the second group while the third group contained four V. albo-atrum isolates. In addition to these three groups, a cluster of six V. nigrescens isolates was observed. However, the genetic distances between the isolates of V. nigrescens were much higher than those between members in the other groups and the bootstrap value for the V. nigrescens cluster was subsequently low. Four isolates classified as V. tricorpus were highly diverse and did not cluster together. Analysis of molecular variance revealed that the isolates of V. longisporum were separated into four subgroups, based on geographic origin. The study furthermore shows that AFLP is a suitable method for studying population structure in Verticillium.     

Analysis of Mycosporine-Like Amino Acids in Selected Algae and Cyanobacteria by Hydrophilic Interaction Liquid Chromatography and a Novel MAA from the Red Alga Catenella repens

Mycosporine-like amino acids (MAAs), a group of small secondary metabolites found in algae, cyanobacteria, lichens and fungi, have become ecologically and pharmacologically relevant because of their pronounced UV-absorbing and photo-protective potential. Their analytical characterization is generally achieved by reversed phase HPLC and the compounds are often quantified based on molar extinction coefficients. As an alternative approach, in our study a fully validated hydrophilic interaction liquid chromatography (HILIC) method is presented. It enables the precise quantification of several analytes with adequate retention times in a single run, and can be coupled directly to MS. Excellent linear correlation coefficients (R² > 0.9991) were obtained, with limit of detection (LOD) values ranging from 0.16 to 0.43 µg/mL. Furthermore, the assay was found to be accurate (recovery rates from 89.8% to 104.1%) and precise (intra-day precision: 5.6%, inter-day precision ≤6.6%). Several algae were assayed for their content of known MAAs like porphyra-334, shinorine, and palythine. Liquid chromatography-mass spectrometry (LC-MS) data indicated a novel compound in some of them, which could be isolated from the marine species Catenella repens and structurally elucidated by nuclear magnetic resonance spectroscopy (NMR) as (E)-3-hydroxy-2-((5-hydroxy-5-(hydroxymethyl)-2-methoxy-3-((2-sulfoethyl)amino)cyclohex-2-en-1-ylidene)amino) propanoic acid, a novel MAA called catenelline.     

The Influence of Organic and Integrated Production on Nutritional,Sensory and Agricultural Aspects of Vegetable Raw Materials for Food Production

In a six-year crop rotation trial organically and integrated grown vegetables were produced according to current good agricultural practices, taking quality and quantity aspects into consideration. The raw materials assessed focussed on materials used for industrial food production. Nutritional, sensory and agricultural aspects were evaluated. Carrot, cabbage, onion, pea and potato are possible to grow organically for industrial purposes. Depending on crop, the yield was lower (65-90%) for organically grown compared to integrated grown. Cultivation of organic spinach and dill turned out to be difficult due to problems with weed and discoloration. The chemical analyses included pesticide residues, nitrate, glycoalkaloid, dry matter, vitamin C and 25 different minerals and trace elements. Overall, the organically grown crops had higher dry matter content than the integrated grown. However, when examining the data for the different crops contradictory results were noted. No significant differences due to growing system were noticed for vitamin C and the other nutrients except for 4 of the trace elements. The growing system did not influence the sensory properties.     

Classification of cassava into ‘bitter’ and ‘cool’ in Malawi: From farmers' perception to characterisation by molecular markers

Cassava roots, a major food in Africa, contain cyanogenic glucosides that may cause toxic effects. Malawian women farmers considered fields of seemingly similar cassava plants to be mixes of both ‘cool’ and ‘bitter’ cultivars. They regard roots from ‘cool’ cultivars as non-toxic. Roots of ‘bitter’ were considered to require extensive traditional processing done by women to be safe for consumption. But curiously, these women farmers preferred ‘bitter’ cultivars since toxicity confers protection against theft, which was a serious threat to the food security of their families. We studied how well these farmers comprehend the effects of genetic variations in cassava when dealing with cyanogenesis in this complex system. Using molecular markers we show that most plants farmers identified as belonging to a particular named cultivar had a genotype typical of that cultivar. Farmers' ethno-classification into ‘cool’ and ‘bitter’ cultivars corresponded to a genetic sub-division of the typical genotypes of the most common cultivars, with four-fold higher cyanogenic glucoside levels in the bitter cultivars. Examining morphology, farmers distinguished genotypes better than did the investigators when using a standard botanical key. Undoubtedly, these women farmers grasp sufficiently the genetic diversity of cassava with regard to cyanogenesis to simultaneously benefit from it and avoid its dangers. Consequently, acyanogenic cassava – the breeding of which is an announced good of some cassava genetic improvement programmes – is not a priority to these farmers. Advances in molecular genetics can help improve food supply in Africa by rapid micropropagation, marker assisted breeding and introduction of transgenic varieties, but can also help to elucidate tropical small-scale farmers' needs and skills. This revised version was published online in August 2006 with corrections to the Cover Date.     

Intermediate landscape disturbance maximizes metapopulation density

The viability of metapopulations in fragmented landscapes has become a central theme in conservation biology. Landscape fragmentation is increasingly recognized as a dynamical process: in many situations, the quality of local habitats must be expected to undergo continual changes. Here we assess the implications of such recurrent local disturbances for the equilibrium density of metapopulations. Using a spatially explicit lattice model in which the considered metapopulation as well as the underlying landscape pattern change dynamically, we show that equilibrium metapopulation density is maximized at intermediate frequencies of local landscape disturbance. On both sides around this maximum, the metapopulation may go extinct. We show how the position and shape of the intermediate viability maximum is responding to changes in the landscape’s overall habitat quality and the population’s propensity for local extinction. We interpret our findings in terms of a dual effect of intensified landscape disturbances, which on the one hand exterminate local populations and on the other hand enhance a metapopulation’s capacity for spreading between habitat clusters.     

Bleeding of Farmed Atlantic Cod: Residual Blood,Color, and Quality Attributes of Pre- and Postrigor Fillets as Affected by Perimortem Stress and Different Bleeding Methods

The main objectives of the study were to (a) assess the effect of perimortem stress on blood drainage, (b) compare the efficiency of gill cutting and direct gutting as bleeding methods, and (c) compare pre- and postrigor filleting strategies for presence of residual blood in Atlantic cod fillets. Anesthetized cod had significantly higher drainage of blood compared to stressed fish. Nevertheless, the visual assessments of residual blood were not affected by stress or bleeding method. Some minor, but significant, differences between pre- and postrigor fillets were found. Stressed fish were initially less light in color than those unstressed, but after ice storage there was no noticeable difference. However, initially, the largest difference in fillet color was due to different bleeding methods. Fillets cut from fish subjected to gill cutting were lighter and less red in color than those bled by direct gutting. After 7 and 21 days of storage, the color differences observed in fresh and salted fillets, respectively, were mainly due to the different rigor status when they were filleted, indicating that fillets cut postrigor were superior to fillets cut prerigor. Prerigor filleting resulted in lower water holding capacity after ice storage in anesthetized, direct gutted cod.     

Solid-phase extraction of pesticides from surface water using discs,bulk sorbents and supercritical fluid extraction (SFE)

Extraction of pesticides from river water samples using the classical liquid-liquid extraction (LLE) approach is a labour-intensive procedure that requires relatively large volumes of solvent. With the solid-phase extraction (SPE) technique using membranes, the volumes of solvent are reduced. However, it is necessary to pre-filter and often also to acidify the river water to pH < 2 to prevent plugging of the capillary pores. This low pH may decompose some pesticides and therefore a fast and inexpensive technique with bulk sorbent and without prefiltration and acidification was developed. To reduce the solvent volumes further, extractions of discs and bulk sorbent were performed with supercritical fluids (SF). Seven pesticides with different polarities were studied. Mean recoveries from the bulk sorbent and the discs were of the same order (61%). However, recoveries from the latter improved to 82% with SFE of the discs, providing results comparable to LLE (94%) recovery.     

Acrylamide and N-methylolacrylamide poisoning in a herd of Charolais crossbreed cattle

Seven beef cattle from a herd accidentally exposed to acrylamide and N-methylolacrylamide while grazing were observed for eight months. They showed clinical signs of impaired nerve function, mainly in the hindlegs, with varying degrees of weakness and ataxia. The animals were irritable, nervous and hypersensitive to touch. Both pupils of the most badly affected animal were dilated and it had poor pupillary light responses; it also showed signs of axonal neuropathy. Selected haematological and clinical chemistry variables were normal. The severity of the neurological signs was correlated with the concentrations of haemoglobin adducts of acrylamides. The animals recovered substantially after their exposure. The gestations of four of the animals which were in calf proceeded normally.     

Effects of topical application of three pyrethroid insecticides on spontaneous activity in sensory nerves of Periplaneta americana and Blabera craniifer

Electrophysiological techniques were used to analyse the effects, on the activity in the sensory nerves of Periplaneta americana and Blabera craniifer, after topical applications of three different pyrethroid insecticides: deltamethrin, (1R)-tetramethrin, and 5-benzyl-3-furylmethyl (E)-(1R)-cis-2,2-dimethyl-3-(2-oxothiolan-3-ylidenemethyl)cyclopropanecarboxylate (RU-15525; ‘Kadethrin’). The insecticides were applied: (a) to the cercus, and the activity was recorded on the cercal nerve and on the central nervous connectives; or (b) to the last segments of the metathoracic leg, and the corresponding activity was recorded on the crural nerve. In both cases, several categories of receptor were affected but the major part of the recorded activity came from the mechanoreceptors, the functional properties of which were monitored throughout the experiments. In most cases, topical application of small amounts of pyrethroid were found to induce an increase in the spontaneous activity, followed by decrease in excitability. Characteristic differences were, however, found between the three compounds and the effects were concentration- and solvent-dependent. The results were consistent with those obtained from experiments on the sense organs of vertebrates but did not support the idea that the effectiveness of pyrethroid insecticides with an α-cyanobenzyl group, compared with those without an α-cyanobenzyl group, originates from a greater effect on the peripheral sensory receptors.