Mucormycosis is a rare and opportunistic infection caused by fungi belonging to the order Mucorales.Recent reports have demonstrated an increasing incidence of mucormycosis, which is frequently lethal, especially in patients suffering from severe underlying conditions such as immunodeficiency. In addition, even though conventional mycology and histopathology assays allow for the identification of Mucorales, they often fail in offering a species-specific diagnosis. Due to the lack of other laboratory tests, a precise identification of these molds is thus notoriously difficult. In this study we aimed to develop a molecular biology tool to identify the main Mucorales involved in human pathology. A PCR strategy selectively amplifies genomic DNA from molds belonging to the genera Absidia, Mucor, Rhizopus, and Rhizomucor, excluding human DNA and DNA from other filamentous fungi and yeasts. A subsequent digestion step identified the Mucorales at genus and species level. This technique was validated using both fungal cultures and retrospective analyses of clinical samples. By enabling a rapid and precise identification of Mucorales strains in infected patients, this PCR-restriction fragment length polymorphism-based method should help clinicians to decide on the appropriate treatment, consequently decreasing the mortality of mucormycosis.

Stock cultures of Xanthomonas campestris NRRL B-1459 require special attention to maintenance and propagation to assure consistent production in good yields of the extracellular polysaccharide xanthan. Under customary conditions of propagative maintenance on agar slants, variant colony types develop that are smaller in size than the normal type. The rate of regression of the normal to the variant forms was diminished when the D-glucose content of the stock medium was sufficient to avoid depletion during storage and when transfer to fresh medium was reduced to 14-day intervals. Under conditions for polysaccharide production, the normal large-colony type gives crude culture liquors after 48 h of 7000 centipoise (cp) viscosity; the predominant variant form gives only 4000 cp. On the basis of 2.1% initial D-glucose, biopolymer yields for the normal and variant strains were 62 and 43%, respectively. Polysaccharide produced by the variant (small-colony type) differs adversely in solution properties from that of the parent strain (large-colony type); it differs also in its lower content of pyruvic acid and O-acetyl substituents. The molar ratios of constituent sugars (D-glucose, D-mannose, and D-glucuronic acid), however, were identical in polysaccharides with the normal and variant strains. Exclusion of colonial variants from fermentations is prerequisite to production of xanthan optimum in properties and yield.

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SummaryXanthan gum is a heteropolysaccharide synthesized by Xanrhomonas cumpestris NRRL B-1459 and is composed of D-glucose, D-mannose, and D-glucuronic acid, in addition to acetic and pyruvic acids. Different amounts of pyruvic acid ketal are found in various preparations which can influence the viscosities of dilute xanthan solutions. Polysaccharide production on synthetic media was studied in small-scale fermentors. Fermentation conditions were established for production of both high and low pyruvic acid gums (about 4 and 2% pyruvic acid, respectively). Low nitrogen [O. 1% (NH4),HP04)] and air (0.25 vol/liter/min) levels favor production of low pyruvate gum; increasing (NH,),HPOI to 0.15%, adding K2HP04, and increasing the air flow to 1.5 vol/liter/min favored production of normal gum. Both processes gave xanthan yields of 50 to 6096, based on 2.5% initial D-glUCOSe substrate, in two to three days. Differences in pyruvic acid content and in the quantity of gum produced under a given set of conditions were attributed to strain variability. Substrains were isolated that have desirable characteristics for production of xanthan gum; i.e., the ability to give good yields of high-pyruvate gum when grown on both complex and synthetic media.