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Comparison of traditional and molecular methods for typing nontoxigenic strains of Corynebacterium diphtheriae.

Thirty-eight nontoxigenic strains of Corynebacterium diphtheriae isolated between 1987 and 1992 from clinical specimens of French patients were typed by biotyping, antibiograms, bacteriophage typing, ribotyping, and restriction analysis by pulsed-field gel electrophoresis (PFGE). Excellent correlation occurred between the genotypes defined by PFGE SfiI profiles or by ribotype BstEII profiles. Genotyping revealed seven genotype patterns among the 26 biotype mitis isolates, five among the nine biotype gravis isolates, and three among the three biotype belfanti isolates. Phage typing was nonreactive for nine of the 38 isolates. A combination of all the typing methods led to the identification of 19 different types of Corynebacterium diphtheriae.

Anti-Bacterial Agents↗

Bacteriophage-encoded type III effectors in Salmonella enterica subspecies 1 serovar Typhimurium.

Salmonella spp. are Gram-negative bacteria which cause infections ranging from mild, self-limiting enterocolitis to systemic (typhoid) disease. Recent work has established that the genetic makeup varies considerably between different Salmonella strains. Phages play an important role in this diversity. In fact, Salmonella has emerged as a prime example for the involvement of virulence factor encoding phages in the emergence of new epidemic strains. Among other virulence factors, Salmonella enterica utilizes two specialized protein secretion systems termed type III secretion systems (TTSS) to deliver effector proteins into host cells which manipulate host cell signaling cascades. These two TTSS and several effectors are encoded within Salmonella pathogenicity islands 1 and 2. Some effectors including SopE, SspH1, SseI and SopE2 are encoded by phages or phage remnants. These phage-encoded effectors seem to be transferred between different Salmonella strains. They have attracted much interest because they might contribute to the evolution of Salmonella spp. Here we will focus on SopEPhi which encodes the SPI-1 effector SopE. It provides an excellent example to illustrate how horizontally transferred effector proteins are integrated into the complex regulatory network of a TTSS in a recipient bacterium. Additional data supporting the hypothesis are presented. This is a prerequisite to allow optimization of the bacterium host cell interaction by reassortment of the phage-encoded effector protein repertoire.

Bacterial Proteins↗

Influence of environmental conditions on infection of Klebsiella pneumoniae by two different types of bacteriophages.

The adsorption and efficiency of plating of bacteriophages FC3-1 and FC3-9 on Klebsiella pneumoniae C3 (serotype O1:K66) cells grown at different pHs and temperatures were quantitated. Bacteriophage FC3-1, with lipopolysaccharide as its bacterial receptor, showed a large decrease in efficiency of plating on bacteria grown at low pH or low temperature. Under the same conditions, no significant decrease in efficiency of plating was found for bacteriophage FC3-9, a phage requiring capsule and lipopolysaccharide for its adsorption and carrying capsule-depolymerizing activity. We demonstrate that K. pneumoniae C3 cells grown at low pH or low temperature have less lipopolysaccharide exposed on their surface. We conclude that this is why lipopolysaccharide-specific phage FC3-1 less efficiently infects bacterial cells grown under those conditions. We propose that bacteriophage FC3-9 efficiently infects bacterial cells grown at low pH or low temperature because its enzymatic activity on the capsule makes lipopolysaccharide available to this phage.

Adsorption↗

Molecular subtyping of Salmonella enteritidis phage type 8 strains from the United States.

Salmonella enteritidis is now the most common serotype of the genus Salmonella reported in the United States. Bacteriophage typing has been helpful for subdividing S. enteritidis strains from different sources in the United States. Most S. enteritidis outbreaks reported were egg related, and the majority of them were caused by strains of phage type 8. To determine whether restriction fragment length polymorphism of the rRNA genes (ribotyping) and of the genomic DNAs from two lysogenic phages from S. enteritidis could be used to discriminate between S. enteritidis phage type 8 strains, we conducted Southern hybridization studies on 24 isolates from different outbreaks and six non-outbreak-associated strains using DNA probes for 16S and 23S rRNA genes and S. enteritidis typing phages 1 and 2 from the Ward typing system (L. R. Ward, J. D. H. de Sa, and B. Rowe, Epidemiol. Infect. 99:291-294, 1987). Of seven restriction endonucleases screened with the probe for rRNA genes, AccI provided the best discrimination between strains; six distinct patterns were observed. AccI ribosomal DNA patterns 1 to 6 were detected among 76.7, 3.3, 6.7, 3.3, 3.3, and 6.7% of isolates tested, respectively. Strains of AccI ribosomal DNA pattern 3 could be further subdivided into two additional patterns by using SmaI. Epidemiologically related strains had identical patterns. No discrimination between strains was achieved by probes for phages 1 and 2. No sequences homologous to the phage I probe were detected among phage type 8 strains, and all strains tested with six restriction enzymes had the same hybridization pattern with the phage 2 probe. These findings demonstrate that ribotyping with AccI and SmaI provides an additional means of discriminating between some phage type 8 strains; however, ribotyping and the phage 2 hybridization results from egg-related outbreak strains support previous findings that these strains are closely related.

Bacterial Typing Techniques↗

[Morphology of Newcastle bacteriophages].

A study was made of morphology of 6 clones of Newcastle bacteriophages of different origin divided into 3 types. Bacteriophage H-18 referred to the III morphological type by the Tikhonenko classification was characterized by a comparatively short process and a head in the form of an isometric polyhedron; H-1, H-5, H-10 and H-17 bacteriophages referred to type V, despite their antigenic difference were morphologically identical: they had a comparatively large head in the form of an elongated polyhedron and a process with a complicated structure ending by a besal plate with 3 indentions originating from it. Bacteriophage H-4 was referred to the IV type and was characterized by a head in the form of an elongated polyhedron and a long curved noncontracting process; in difference from the others it had no basal plate on the end of the process. The revealed morphological peculiarities of the particles of the Newcastle bacteriophages only partially correlated with their division on the basis of serolological properties and the size of the negative colonies.

Bacteriophages↗

A new phage typing scheme for Proteus mirabilis and Proteus vulgaris strains. 1. Morphological analysis.

A new bacteriophage typing set, composed of 22 phages, was established as a tool for differentiation of Proteus strains. All the phages were tailed and included 4 morphological types (A1, A2, B1 and C1). They were classified into the families Myoviridae, Siphoviridae and Podoviridae. From the set, 19 phages had double-stranded DNA and 3 were single-stranded DNA phages.

Bacteriophage Typing↗

Plasmid analysis and phage typing in the study of staphylococcal colonization and disease in newborn infants.

Plasmid analysis, bacteriophage typing and antibiograms were used to detect the source and rate of Staphylococcus aureus colonization and disease in newborn infants in a nursery. Staphylococcal colonization on the nasal mucosa and umbilicus of 140 neonates took place within 12 hours after birth. The colonization rates reached 100% from the fourth day onward and fell to 66.7%, 25.5% and 17.5% by the age of 1 month, 6 months and 1 year, respectively. The incidence of nursery-acquired infection caused by S. aureus was at least 16.4%. All infections were confined to the skin or mucosa and were not serious. It is believed that the S. aureus which colonized and infected the newborn infants mainly originated from the nares and the hands of nurses, because only S. aureus strains from nurses and infected infants had the same phage patterns (52/52A/47/53/75/77/81) and carried 2 plasmids of the same size (1.4 and 1.7 Md), whereas S. aureus isolated from other sources did not have the same phage patterns or plasmid profiles. In view of the results of this study, it is considered that the importance of careful washing of the hands of personnel before and after handling infants should be emphasized.

Bacteriophage Typing↗

Molecular typing of Escherichia coli O157:H7 (H-) isolates from cattle in Japan.

A total of 77 Escherichia coli O157:H7 (H-) isolates from cattle in Japan were investigated by molecular biological methods. Most of these isolates (43 isolates) possessed the stx-2 gene, but not stx1. Fifteen bacteriophage types and 50 pulsed-field gel electrophoresis (PFGE) profiles were observed. One isolate was indistinguishable from the human outbreak strain by these methods. This indicates that cattle must be considered as a possible source of human E. coli O157:H7 infection in Japan.

Animals↗

Epidemiology of Clostridium difficile colonization in newborns: results using a bacteriophage and bacteriocin typing system.

We used a typing system based on bacteriophage and bacteriocin susceptibility to study the epidemiology of Clostridium difficile colonization of newborn infants. C. difficile was found in the stools of 30 (16.0%) of 187 infants who were screened. Increased length of stay in the nursery (P less than .001) and delivery by cesarian section (P less than .001) were associated with higher rates of colonization. The isolates initially detected from the environment and the infants were strain B1811-1700. Strain B1537/Cld7 became the predominant isolate obtained from the infants; positive cultures were also obtained from the environment and the hands of personnel who worked in the nursery and had strain B1537/Cld7. Our results suggest that the infants acquired C. difficile through transfer from the hands of hospital staff.

Bacterial Proteins↗