- Volume 139, Issue 11, 1993

Three-hundred and twenty-six strains of Xanthomonas campestris pv. manihotis from 22 countries were studied to detect and assess genetic and evolutionary relationships within the pathovar. A range of techniques was used for this study including restriction fragment length polymorphism (RFLP) analysis. The probes used for the RFLP analysis were 16 + 23S rRNA genes from Escherichia coli and three restriction fragments from the chromosomal and plasmid DNA of X. campestris pv. manihotis. Analysis of the rRNA probe data showed five RFLP groups whilst the other three probes were used to further sub-divide these groupings. Genetic variability of X. campestris pv. manihotis was pronounced in strains from South America where the host plant originated but was limited in strains from other regions. The results obtained confirm the hypothesis that the pathogen has been introduced only recently to Africa and suggests that African strains have not as yet diversified significantly at the chromosomal level. Our results indicate that rRNA and DNA probes are useful tools for epidemiological studies and in following the genetic evolution of strains.

This paper describes the characterization and taxonomic status of N16, a chlamydial isolate from the respiratory tract of a horse. N16 contains plasmid DNA, has normal elementary body morphology and its inclusions do not stain with iodine. Its major outer-membrane protein (MOMP) gene was completely sequenced and compared with the MOMP genes of Chlamydia pneumoniae, C. psittaci, C. trachomatis and C.pecorum. This analysis revealed that N16 is closely related to the TWAR strain of C. pneumoniae (94·5% and 94·4% DNA homology with TWAR isolates IOL-207 and AR-39 respectively). By comparison, N16 shows between 72·1% and 73·7% DNA homology with C. psittaci strains, 70·9% and 71·1% homology with C. pecorum strains LW613 and 1710S and 69.2% homology with C. trachomatis serotype E. The MOMP gene of N16 shares 93·8% DNA homology with the MOMP gene of a chlamydial isolate KC from the conjunctiva of a koala. Monoclonal antibodies raised to C. pneumoniae IOL-207 and shown to be C. pneumoniae-specific confirmed that N16 was more closely related to C. pneumoniae than to C. psittaci. Thus DNA homology and monoclonal antibody data both suggest that horse chlamydiae, as exemplified by N16, form a new second strain of C. pneumoniae. This species is probably more widespread and diverse than the current literature would suggest.

Out of 22 Enterobacter phages investigated, nine were found to be suitable for phage typing based on their different lytic spectra on 398 strains of Enterobacter spp. isolated from milk powder and other foods. These phages were compared on the basis of morphology, protein composition, restriction endonuclease patterns and DNA-DNA hybridization. Two phages (WS-EP19, WS-EP13) belonged to the Podoviridae family (morphotype C1), and three (WS-EP20, WS-EP26, WS-EP28) were classified as Siphoviridae (morphotype B1). The other four phages were Myoviridae of the morphological groups A1 (WS-EP57) and A2 (WS-EP32, WS-EP94, WS-EP96). SDS-PAGE revealed individual protein profiles for each phage, which corresponded to different restriction enzyme fragment patterns. DNA-DNA hybridization demonstrated the close relationship of phages WS-EP20 and WS-EP26, and of WS-EP94 and WS-EP96. In general, a good correlation was found between groupings obtained with the various methods. The nine phages could be attributed to existing enterobacterial phage species although some differences to the described type phages were observed.

Antigens of Eubacterium species including E. alactolyticum, E. brachy, E. nodatum, E. saburreum, E. timidum, E. yurii subsp. yurii and E. yurii subsp. margaretiae, which have been isolated frequently from periodontal pockets and associated with periodontal diseases, were extracted by ultrasonication from whole bacterial cells. Antigens were also prepared from E. aerofaciens, E. lentum and E. rectale, which have been found in intestinal tracts and infected abscesses in human oral cavities. The antigens of the oral Eubacterium species were compared with antigens from E. limosum, the type species of the genus Eubacterium, by using SDS-PAGE and Western immunoblot assays. SDS-PAGE gels stained with Coomassie brilliant blue indicated that no major peptide bands were common among the Eubacterium species examined. The protein profile patterns were distinctly different from each other. Western immunoblotting reactions with rabbit antisera showed that the Eubacterium species could be clearly distinguished serologically, and that the species-specific antigens were peptide components of ultrasonic extracts from the whole bacterial cells. The present study demonstrates that these Eubacterium species show great heterogeneity in their peptide components and immunological reactions, which may be useful for identification of the Eubacterium species from human oral specimens.