Molecular detection of anthrax spores on animal fibres.

AIMS: To develop a rapid, specific and sensitive diagnostic test for the detection of the spores of Bacillus anthracis on commercial samples of animal fibres (e.g. wool and cashmere). METHODS AND RESULTS: Extraction of DNA from spores using a mechanical disruption method based on bead beating was evaluated but subsequently abandoned as it compromised the sensitivity of the overall protocol. A multiplex PCR and two nested amplification reactions designed for B. anthracis were developed during this study. CONCLUSIONS: A simple selective incubation step in combination with multiplex PCR was found to be more effective than generic DNA extraction coupled to a sensitive nested amplification reaction. SIGNIFICANCE AND IMPACT OF THE STUDY: The rapid diagnostic test could be applied to the analysis of commercial fibre samples for the detection of anthrax as required by health and safety legislation resulting in considerable savings in time and expense.

Assessment of cooked alpaca and llama meats from the statistical analysis of data collected using an 'electronic nose'.

As part of an EU-funded project to assist in developing the production chain of meat from camelids in South America we have investigated the possibility of using an electronic nose to distinguish between the different types of meat of commercial interest. On-site monitoring of freshly cooked camelid meat using a Bloodhound electronic nose has been carried out in Peru and Bolivia. Sampling was carried out using inert, collapsible plastic bags. Linear discriminant analysis of data generated by the electronic nose classified the samples of meat. Some problems experienced in analysing the data relating to sample size are discussed.

Talking rot ... and mildew on textiles.

Nature has evolved some very elegant elementary mechanisms for re-cycling of organic substances in the environment. These processes are normally beneficial. This article describes why microorganisms may attack textiles during production and in use, causing financial losses, and what can be done to prevent this happening.

Biocompatibility of potential wound management products: hydrogen peroxide generation by fungal chitin/chitosans and their effects on the proliferation of murine L929 fibroblasts in culture.

Agaricus bisporus, Fusarium graminearum, Phycomyces blakesleeanus, unbleached and bleached, Rhizomucor miehei, and Rhizopus oryzae were examined as sources of fungal chitin/chitosan. The nitrogen content of the alkalitreated mycelia/sporangiophores obtained after optimization of culture conditions, and of similarly treated A. bisporus stipes, was 2.87, 1.29, 6.27, 6.50, 4.80, and 4.95% w/w, respectively, which relates to an estimated chitin content of 42, 19, 91, 94, 70, and 72%, respectively. The hydrogen peroxide (H2O2)-generating ability of the treated fungal materials after 8 h at pH 7.4 and 37 degrees C decreased in the order R. oryzae > P. blakesleeanus unbleached approximately R. miehi > F. graminearum > A. bisporus > P. blakesleeanus bleached. This did not correlate with estimated chitin content. The effect of these fungal materials on the rate of proliferation of murine L929 fibroblasts in culture also was examined. Both pro- and antiproliferant effects were observed. Significant (P < .05) proproliferant effects were observed on day 6 with R. miehei, R. oryzae, and P. blakesleeanus (unbleached and bleached) at 0.01% w/v. The greatest antiproliferant effect was observed with R. oryzae at 0.05% w/v on day 6 (-63% relative to the control, P < .05; cell viability, 95%). In contrast, A. bisporus failed to affect cell yield significantly at either 0.01 or 0.05% w/v. Addition of catalase to cultures containing R. oryzae or R. miehei at 0.05% w/v failed to abolish the antiproliferant effect on day 3, instead producing a small but significant (P < .05) increase in the effect. Catalase also failed to affect significantly the antiproliferant effect of F. graminearum at 0.05% w/v, but did abolish the proproliferant effect of P. blakesleeanus (unbleached and bleached) on day 3. Overall, our results suggest that the H2O2 being generated by the fungal materials modulates cell proliferation but that this effect is superimposed upon a H2O2-independent antiproliferant effect manifesting itself at the higher concentrations of fungal material. The antiproliferant effect was not attributable to Ca2+, Mg2+, or Fe2+ depletion although chelation of Fe2+ did correlate with H2O2-generating ability. Only P. blakesleeanus appears to lack this antiproliferant activity while retaining H2O2-generating activity. These results may aid the selection of fungal chitin/chitosan for further evaluation as a potential wound management material.

Biocompatibility of potential wound management products: fungal mycelia as a source of chitin/chitosan and their effect on the proliferation of human F1000 fibroblasts in culture.

Aspergillus oryzae, Mucor mucedo, and Phycomyces blakesleeanus cultures were examined as sources of chitin/chitosan. The nitrogen content of the alkali-treated mycelia/sporangiophores of A. oryzae, M. mucedo, and P. blakesleeanus was 2.52, 3.61, and 6.27% w/w, which relates to an estimated chitin content of 37, 52, and 91%, respectively. The effect of these fungal materials on the rate of proliferation of human F1000 fibroblasts in culture was examined. At 0.01% w/v, all three materials exhibited significant (P < .05) proproliferant activity over a period of 13 days. However, at 0.05% w/v, P. blakesleeanus further enhanced cell proliferation, whereas A. oryzae and M. mucedo produced a significant (P < .05) antiproliferant effect. Higher concentrations of P. blakesleeanus (0.1 and 0.5%) caused marked inhibition of F1000 cell proliferation when measured on days 3 and 6. Only the proproliferant effect of these fungal materials appears to correlate to their chitin content. Furthermore, the cytomorphology of the fibroblasts indicated that P. blakesleeanus, and to a lesser extent M. mucedo, possessed cell attractant properties, again correlating with chitin content. If developed for use as wound management materials, the sporangiophores of P. blakesleeanus and the mycelium of M. mucedo could possibly promote the growth of fibroblasts and provide a matrix for their anchorage, thus contributing to the granulation phase of the healing cascade.

Protoplast fusion as a tool for genetic analysis in Cephalosporium acremonium.

Protoplasts of nutritionally complementary strains of Cephalosporium acremonium were fused and plated onto media which supressed the growth of both parents. The regenerating colonies were used for genetic analysis and were found to be of two types, stable haploid recombinants and unstable heterozygotes (aneuploids and/or diploids). Analysis of these colonies provided evidence for eight linkage groups and a relatively high rate of mitotic crossing-over. The gene order for three of the markers on one linkage group was also determined.