Evaluation of PCR,IEF and ELISA techniques for the detection and identification of potato cyst nematodes from field soil samples in England and Wales

Effective management of potato cyst nematodes (PCNs) requires simple, rapid and accurate identification and quantification of field populations. Soil samples from a survey of 484 fields in potato rotations in England and Wales were used to compare the identification and quantification of PCNs using IEF, PCR, ELISA and bait plant tests. The cyst counts and bait plant test revealed that 64.3% of field samples contained PCNs. Bait plant tests increased the detection rate of PCNs in field samples by 4–6.4%. This means that some infestations are cryptic and would not normally be detected by standard counts. IEF, PCR and ELISA methods distinguished between Globodera rostochiensis and G pallida and were able to register mixed populations; however they were not in full agreement. All methods suggested that G pallida is the dominant species in the field samples tested. The PCR results indicated that 66% of field samples contained pure G pallida, 8% contained pure G rostochiensis and 26% contained mixtures of the two species. Estimates of the relative process times taken per sample in the PCR, IEF and ELISA techniques are given. © 2001 Society of Chemical Industry     

The in vitro motility response to various anthelmintics of third-stage larvae of Oesophagostomum spp. from pigs

The in vitro activities of thiabendazole, levamisole, pyrantel, morantel and ivermectin against Oesophagostomum spp., the nodular worm of pigs, were determined and compared. The study was carried out using isolates of O. dentatum and O. quadrispinulatum, which had been defined in vivo. Infective larvae were exposed to the anthelmintics for 24 h and then placed in a micromotility meter. All the treatments significantly reduced the motility of the ensheathed L₃ larvae, but the micromotility meter was not able to differentiate between anthelmintic resistant and anthelmintic susceptible isolates.     

Genetic typing of bovine viral diarrhoea virus: most Slovenian isolates are of genotypes 1d and 1f

A selection of 43 bovine viral diarrhoea viruses isolated from mainly persistently infected cattle on 23 Slovenian farms between 1997 and 2001 were characterised genetically. Viral RNA was extracted from infected cell cultures, reverse transcribed and amplified by PCR with primers targeting the 5'-UTR and the N(pro) gene, followed by direct sequencing of purified PCR products obtained for both genomic regions. The N(pro) sequences provided the best genetic resolution, and gave also higher statistical support for phylogenetic classification of the viruses. Thirty-eight of the Slovenian isolates were of genetic subtypes 1d and 1f, four were 1b, and one subtype 1g. No BVDV type 2 viruses were found. This genetic prevalence matched those previously reported for neighbouring countries, as opposed to findings reported for more distant European countries, e.g. France, Spain and the UK. From eight cattle herds several virus isolates were analysed; with one exception all isolates from each herd were of the same genetic group. Extended sequencing of the N(pro) and part of the C gene of virus isolates with identical 5'-UTR sequences allowed differentiation between isolates obtained at different times from one herd.     

On the fracture morphology in wood

Summary Structural changes in spruce and aspen wood samples subjected to axial compression were examined by means of the scanning electron microscope. For comparison, macroscopic model experiments were carried out on tube-shaped samples made of paper so as to represent segments of fibriform xylem cells. The results show that there are fracture patterns characteristic of wood in general and others characteristic of the species of wood. The phenomena characteristic of wood in general are prevalent at the cellular level and in the fracture behaviour of the cell wall layers and perforations. The characteristics of the various species of wood dominate at the level of the growth rings and are decisively influenced by the composition and arrangement of the tissues. The model experiments show that the failure morphology of the individual cells of the xylem may be explained to some extent by their geometry.     

Chemical composition of hemicelluloses as a factor participating in the substrate specificity of wood-destroying fungi

Mycelium of twenty species of wood-destroying fungi was cultivated in nutrient solutions with one of the following sugars as the only carbon source: glucose, arabinose, galactose, mannose and xylose. The growth of the species occurring in nature only on the wood of Angiosperms was equal or lower in the solutions with galactose and mannose than in the solution with glucose. The utilization of mannose was practically the same as that of xylose. The species which are specialized in nature in the wood of conifers grow better in the solutions with galactose and mannose, than in the solution with glucose; likewise mannose is beter utilized than xylose. Fungi attacking both kinds of wood do not exhibit any dependence on the utilization of the sugars used in these investigations.     

Technological properties and fire performance of medium density fibreboard (MDF) treated with selected polyphosphate-based fire retardants

ABSTRACT

The objective of the work was to evaluate the efficacy of two new polyphosphate-based fire retardants (FRs) and one commercial product named Siriono^® on the fire performance and physical–mechanical properties of medium density fibreboard (MDF) fabricated in the laboratory from Scots pine (Pinus sylvestris L.) wood. The fibres were treated with aqueous solutions of fire retardants, at 12% loading (dry salt on dry wood), and bonded with a melamine urea formaldehyde (MUF) adhesive. The physical and mechanical properties of panels were assessed using the European standards, whereas their fire performance was evaluated using an in-house method and the Cone calorimeter. In overall, the chemicals added enhanced the fire and smoke properties of the panels to varying degrees. Critical FR parameters such as peak heat release rate (peak HRR), total heat release (THR) and total smoke production (TSP) were significantly improved in the FR-treated panels, as exhibited in cone calorimeter tests. However, the internal bond strength of treated panels largely decreased by the addition of fire retardants, while thickness swell and water absorption negatively affected to a significant extent. In contrast, the formaldehyde release of the panels was considerably decreased at the E1 class level, with the incorporation of the polyphosphate-based additives.     

Localization of CD9 Molecule on Bull Spermatozoa: Its Involvement in the Sperm–Egg Interaction

Tetraspanin CD9 is one of the egg membrane proteins known to be essential in fertilization process. The presence and localization of CD9 molecule in spermatozoa and its possible function in reproduction are still unclear. In our study, we describe the localization of CD9 on bull spermatozoa. In the immunofluorescence assay, the positive signal has been observed in the high proportion of sperm cells as a fine grains either on the apical part or through the entire anterior region of sperm head. CD9 recognized by monoclonal antibody IVA‐50 was detected on freshly ejaculated (83.4 ± 3.7%) and frozen‐thawed (84.3 ± 2.3%) sperm. The same reaction pattern was observed on sperm capacitated for 1 h, 2 h, 3 h and 4 h (83.6 ± 2.0%; 84.0 ± 1.5%; 85.7 ± 0.8%; 77.5 ± 10.8%). The presence of CD9 exclusively on plasma membrane of the bovine sperm has been detected by Western blot analysis of the protein fractions after the discontinuous sucrose gradient fractionation of the bull sperm. Moreover, probable role of the sperm CD9 molecule in fertilization process of cattle has been suggested as sperm treatment with anti‐CD9 antibody significantly reduced (by 25%, p ≤ 0.001) the number of fertilized oocytes compared to control group in fertilization assay in vitro.     

Chigger mites (Acari: Trombiculidae) from wild birds in Costa Rica, with a description of three new species

Three new species of chigger mites, Eutrombicula costaricensis sp. n., Eutrombicula passerinoruni sp. n., and Eutrombicula hectochaeta sp. n. are described from wild birds from Costa Rica. Two species, Eutrombicula pacae (Floch et Fauran, 1957) and Parasecia findata (Brennan, 1969), are recorded for the first time in Costa Rica and on new host species. Data on the distribution of Blankaartia sinnamaryi (Floch et Fauran, 1956) in Costa Rica are also reported.     

Can the structure of dormant cambium and the widths of phloem and xylem increments be used as indicators for tree vitality?

We investigated the structure and width of the dormant cambium and of the increments of phloem and xylem of Quercus robur to estimate their potential as indicators for tree vitality. The samples were taken from three woodlands, two in Slovenia [Krakovo forest (KRA) and Murska Suma (MUS)] and one in Croatia [Kobiljak (KOB)], with reported tree decline. The number of dormant cells seems to reflect the initial capacity of the cambium to accomplish cell division. With the exception of two trees at KRA, cell production was always higher on the xylem side than on the phloem side. The annual phloem increments were narrower, less variable among trees and with clear lower and upper limits. With increased cambial cell productivity, the share of the xylem in the total annual radial increment increased following a curvilinear function. In trees with an annual radial increment >3.5 mm, the xylem size represented more than 90 % of the total radial growth. The anatomical variables analyzed show that the most limiting environmental conditions seem to prevail at KRA, whereas the conditions at MUS seem to be most favorable in terms of radial growth. Analysis of the width and structure of xylem and phloem increments, the number of dormant cambial cells and their inter-relationships can provide additional information on the vitality of oaks.