J.B. Woolcock

Experimental infection of mice with Rhodococcus equi: Differences in virulence between strains

The growth kinetics in outbred mice of clinical and environmental isolates of Rhodococcus equi were followed by serial bacterial enumeration of organ homogenates. Clinical isolates multiplied until Day 4 before being progressively cleared, but could still be recovered from the liver at 3-4 weeks post-infection. Intravenous inoculation of clinical strains was associated with histopathological responses very similar to those elicited by intravenous infection with various facultative intracellular parasites. Whereas lesions in mice and foals at 7-9 days following respiratory infection are those of severe bronchopneumonia with massive consolidation, a week later the patterns of host response have diverged as the murine lesions resolve. The type strain, NCTC 1621 and 4-6 environmental isolates were eliminated without prior multiplication and these strains caused negligible lesions. The two environmental strains which behaved as the clinical strains were recovered from a stud with an R. equi problem. No association of colonial morphology of R. equi with virulence was apparent.

Early events associated with experimental infection of the murine lung with Rhodococcus equi

Pneumonia due to Rhodococcus equi was induced in the murine lung by deposition of a known dose of organisms. From serial estimations of bacterial numbers in the lungs of inoculated mice, analysis of the cellular composition of bronchoalveolar lavage fluid and morphological examination of the lungs, events in the host-parasite interaction were followed until day 7. Early bacterial clearance from the lung was dose-dependent but was not sustained. A proportion of the inoculated R. equi was susceptible to the early nonspecific phagocytic cell response, and the contribution of neutrophils to bacterial clearance appeared largely limited to the first 24 hours. A substantial fraction of the organisms survived in the alveoli, probably within macrophages. The contribution phagocytes make to resistance against R. equi is similar to that which prevails in infection with Listeria monocytogenes.

Some problems associated with the identification of Corynebacterium equi

One hundred strains of Corynebacterium equi were examined using standard laboratory procedures. Variations in colonial morphology were noted. The ability to reduce nitrate and to produce urease was shown to be valuable for confirmation of identification of C. equi. Production of hydrogen sulphide was a variable characteristic of C. equi. It is concluded that for those situations where the source of the organism gives no clue as to its identify, positive identification of C. equi may be difficult because of variability in colonial appearance and biochemical reactions.

The effect of immunosuppression on resistance to Rhodococcus equi in mice

Rhodococcus equi, a natural pathogen of horses, produces lesions in mice following experimental infection. The effect of various immunosuppressing agents on the sequential development of these lesions has been assessed by measuring the growth of R. equi following intravenous or intranasal challenge and by histological examination. Cyclophosphamide treatment of mice, challenged intranasally, resulted in the development of lesions not unlike that seen in experimental and natural infection in foals. Cortisone acetate also impaired bacterial clearance from the lungs and affected the accumulation of mononuclear cells at infective foci. Most of the agents chosen to impair macrophage function failed to affect the resistance of mice to R. equi. Carbon, carrageenan and silica failed to alter significantly the growth kinetics of R. equi. Dextran sulphate depressed the rate of pulmonary clearance of organisms and affected the ability of animals to eliminate R. equi following rechallenge. Overall, these results support other evidence that cell mediated immunity is involved in host resistance to R. equi and that activated macrophages play a role in acquired immunity to this organism.

Phage types of Australian isolates of Salmonella enterica subsp. enterica serovar Virchow

Australian isolates (79) of Salmonella enterica subsp. enterica serovar Virchow (Salmonella Virchow) were characterized by phage typing. Thirteen phage types were identified, of which phage type (PT) 8, representing 54 of 79 isolates, was predominant, as it had been in England and Wales up to 1994 when it was replaced by PT26. Other phage types identified in Australia were distinct from those observed in England and Wales. This suggests that PT8 may be a global phage type, while others may be distinct to particular geographical regions.

Plasmid analysis of Australian strains of Salmonella enteritidis

Using an in‐well lysis technique, 73 Australian strains of Salmonella enteritidis were shown to possess a large plasmid, similar in size to that possessed by a reference phage type 4 strain. Restriction analysis of the large plasmid from nine strains using EcoRI, HindIII and PstI suggested that these plasmids are similar to or the same as the 38 MDa plasmid described in strains of this species from other parts of the world.

Lipopolysaccharide expression and virulence in mice of Australian isolates of Salmonella enteritidis

The lipopolysaccharide (LPS) of 54 Australian isolates, nine isolates acquired or isolated overseas, and two reference strains of Salmonella enteritidis was studied to assess its relation to pathogenicity. LPS was extracted by proteinase K digestion of whole cells, and analysed by polyacrylamide gel electrophoresis. All isolates possessed an LPS structure identical to that of a reference strain of Salm. enteritidis phage type 4. Representative strains of the clinically prevalent phage types 4, 14 and 26, which express long chain LPS, were assessed for their pathogenicity in mice. Salmonella enteritidis phage type 4 produced a lethal infection in BALB/c mice, but not in C3H/HeJ or Quackenbush (outbred) strains. Phage types 14 and 26 did not produce an obvious infection in any mice, suggesting Australian strains of phage type 4 are more virulent than phage types 14 and 26.

API ZYM for identification of Corynebacterium equi

Summary The API ZYM system has been evaluated for use in identification of C. equi . One hundred strains were tested, and compared with representatives of coryneform and related taxa. The system is rapid and reliable and gives a pattern of reactions of C. equi which is consistent and characteristic.

Sensitivity of Australian isolates of Salmonella enteritidis to nitrofurantoin and furazolidone

Susceptibility of 66 and 62 Australian isolates of Salmonella enteritidis to nitrofurantoin and furazolidone, respectively, was determined. Most isolates were susceptible to both antibiotics. Cross-resistance was low among all isolates, but higher among the subset of phage type 4 isolates. These results contrast directly with those of a previous study (Rampling, A., Upson, R. and Brown, D.F.J. (1990) J. Antimicrob. Chemother., 25: 285-290). Sensitivity among Australian isolates of S. enteritidis does to some extent, support the contention that furans may have played a role in the selection and enhanced colonisation of poultry by Salmonella enteritidis in Britain. Furthermore, nitrofurantoin should not be used as a selective agent in the isolation of Salmonella enteritidis.

Multilocus enzyme electrophoretic (MEE) analysis of Australian isolates of Salmonella Enteritidis

Seventy-three Australian isolates of Salmonella Enteritidis (SE) were analysed by multilocus enzyme electrophoresis (MEE) using a polyacrylamide gel system. Analysis of 11 enzyme loci identified eight electrophoretic types (ETs), with 61 of the isolates assigned to ET1, and 72 isolates considered to represent a clonal lineage. Representative isolates of each of the Australian ETs were then compared with isolates from England, Germany and the United States, using a starch gel system and 13 enzyme loci. The overseas isolates formed a single ET with representatives of the major Australian ET. It is concluded that Australian isolates of SE are closely related genetically to those from countries in which egg-borne transmission is common.