Bacterial Blight and Dieback of Eucalyptus Species, Hybrids, and Clones in South Africa.

During 1998, a new disease appeared on trees representing a Eucalyptus grandis × E. nitens (GN) hybrid in a nursery in KwaZulu/Natal. The disease has subsequently spread to other Eucalyptus species, hybrids, and clones in nurseries and plantations throughout South Africa. Typical symptoms of the disease include dieback of young shoots and leaf blight. This ultimately leads to stunting of trees. The objective of this study was to isolate and identify the causal agent of the disease. A bacterium was consistently isolated from infected tissue. Pathogenicity tests were undertaken with a range of bacterial strains. Four pathogenic strains were selected from different geographical regions and Eucalyptus hosts for further study. The bacterium causing Eucalyptus leaf and shoot blight is gram negative and rod-shaped, varying in size from 0.5 to 0.75 µm wide and 1.0 to 2.0 µm long. Colonies of this bacterium have a yellow pigment. The results from the Biolog tests identified the bacterium as Pantoea agglomerans with a similarity index of 0.315. The 16S rDNA sequences of the purported Pantoea sp. were compared with those of other related Enterobacteriaceae from GenBank/EMBL. Phylogenetic analysis using PAUP revealed that the isolates group together with P. agglomerans, P. ananatis, and P. stewartii subsp. stewartii. The fatty acid profiles and phenotypic characteristics of the new pathogen are similar to P. ananatis, and % G + C is within the range of this species. DNA:DNA hybridization between the four strains and the type strain of P. ananatis conclusively showed that the bacterium causing blight and dieback of Eucalyptus in South Africa belongs to this species. This is the first report in which P. ananatis has been found as a causal agent of a disease on Eucalyptus.

BTEX catabolism interactions in a toluene-acclimatized biofilter.

BTEX substrate interactions for a toluene-acclimatized biofilter consortium were investigated. Benzene, ethylbenzene, o-xylene, m-xylene and p-xylene removal efficiencies were determined at a loading rate of 18.07 g m(-3) h(-1) and retention times of 0.5-3.0 min. This was also repeated for toluene in a 1:1 (m/m) ratio mixture (toluene: benzene, ethylbenzene, or xylene ) with each of the other compounds individually to obtain a final total loading of 18.07 g m(-3) h(-1). The results obtained were modelled using Michaelis-Menten kinetics and an explicit finite difference scheme to generate vmax and Km parameters. The Vmax/Km ratio (a measure of the catalytic efficiency, or biodegradation capacity, of the reactor) was used to quantify substrate interactions occurring within the biofilter reactor without the need for free-cell suspended and monoculture experimentation. Toluene was found to enhance the catalytic efficiency of the reactor for p-xylene, while catabolism of all the other compounds was inhibited competitively by the presence of toluene. The toluene-acclimatized biofilter was also able to degrade all of the other BTEX compounds, even in the absence of toluene. The catalytic efficiency of the reactor for compounds other than toluene was in the order: ethylbenzene > benzene > o-xylene > m-xylene>p-xylene. The catalytic efficiency for toluene was reduced by the presence of all other tested BTEX compounds, with the greatest inhibitory effect being caused by the presence of benzene, while o-xylene and p-xylene caused the least inhibitory effect. This work illustrated that substrate interactions can be determined directly from biofilter reactor results without the need for free-cell and monoculture experimentation.

A novel oxylipin-associated 'ghosting' phenomenon in yeast flocculation.

Research on the distribution of oxylipins (3-hydroxy fatty acids) in flocculant strains of the yeast Saccharomyces cerevisiae led to the uncovering of a novel 'ghosting' phenomenon observed during assumed lectin-mediated aggregation. We found that intracellular oxylipin-containing osmiophilic layers migrate through yeast cell walls in a 'ghostlike' fashion without visually affecting the cell wall structure or the layers. This migration resulted in the binding of these layers to cell walls of adjacent cells. Consequently, 'ghosting' seems a prerequisite for flocculation to occur. However, 'ghosting' alone may not be sufficient to ensure flocculation.

Identification of classical Propionibacterium species using 16S rDNA-restriction fragment length polymorphisms.

The phenotypic identification of the classical propionibacteria is essentially still problematic and alternative techniques for the identification of the various species are required. A rapid and sensitive technique for the routine identification of the classical propionibacteria, based on the amplification of 16S rRNA genes using the polymerase chain reaction and the subsequent restriction endonuclease digestion of the PCR products, was previously described. Although this technique enabled differentiation between the various classical species examined it was only evaluated on a limited number of type and reference strains. During this study, the taxonomic relationship between 135 Propionibacterium strains from diverse ecological niches, representing four classical species was investigated using this PCR/RFLP technique. Visual differentiation between the classical Propionibacterium was possible after restriction endonuclease digestion of the PCR products obtained using primers 16sP1-16sP4 and 16sP3-16sP4 with the restriction endonucleases HaeIII, AluI and HpaIII, respectively. With the exception of strains independently identified as "P. rubrum" and "P. sanguineum", the results of this study confirm the consolidation of the "old" species into the various classical species as they currently exist. It was therefore concluded that the PCR/RFLP protocol is suitable for the rapid and routine identification of the classical propionibacteria.

Propionibacterium species diversity in Leerdammer cheese.

Thirteen 'classical' Propionibacterium strains, isolated from Leerdammer cheese samples, using three different media were characterized phenotypically. The phenotypic data of 74 tests, conducted on 27 propionibacteria, including four type, 10 reference strains and the 13 cheese isolates were analysed by numerical taxonomical techniques, using the simple matching coefficient and single linkage cluster analysis. All the strains were grouped in four major clusters, with a final linkage at the 81% S-level. The clusters were equated with the 'classical' P. acidipropionici, P. freudenreichii, P. jensenii and P. thoenii species. The species were identified by relating them to specific type strains and by comparison of phenotypic characteristics. Differential characteristics of each cluster were determined. Strains of P. acidipropionici, P. freudenreichii and P. jensenii, but no P. thoenii strains were isolated from the Leerdammer cheese samples. No 'cutaneous' propionibacteria were isolated. The largest cluster, representing 46% of the cheese isolates was equated with P. jensenii. Various red/brown pigmented strains, which could be identified as the old 'P. rubrum' species were isolated from the cheese. These strains were, however, phenotypically identified as P. jensenii and also grouped in the P. jensenii cluster.