Graham Selby Wilson: 10 September 1895-5 April 1987.
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Biomedical subjects
Publications and source records attributed to E S Anderson.
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Since 1969 strains of Salmonella wien that are resistant to multiple antibacterial drugs have caused widespread epidemics of enteritis in Europe and North Africa. Of 113 British strains examined between January 1970 and January 1977, 67 were multi-resistant. These strains and 22 strains from six other countries were examined to determine their plasmid content. Two plasmids were found in the vast majority of strains: an FIme factor, conferring resistance to ampicillin, chloramphenicol, kanamycin, streptomycin, spectinomycin, sulfonamides, and tetracyclines (ACKSSpSuT), and a nonautotransferring plasmid of resistance type ASSu. The FIme plasmids have dual incompatibility: they are incompatible with group FI factors and with the MP10 plasmid of Salmonella typhimurium, which belongs to a separate group. Other plasmids found in S. wien included principally a ColIa factor and an autotransferring plasmid that codes for ampicillin resistance and belongs to compatibility group I2. The similarity in plasmid content of strains isolated in widely separated areas suggests that they have a clonal origin.
A simple gel electrophoresis method has been described for the detection of plasmid DNA in bacteria (Meyers et al., 1976). We investigated further the problems encountered in using this method for the analysis of plasmids in wild enterobacterial strains. The migration of open circular and linear plasmid DNA was examined, since these forms sometimes caused difficulty in the interpretation of the plasmid content of uncharacterized strains. Electrophoresis at different agarose concentrations was employed to resolve clearly plasmid DNA from the chromosomal DNA fragments in the crude preparations. Dissociation of some plasmids occurs in Salmonella typhimurium, and this was detected by electrophoresis. The technique was applied to the study of drug-resistant strains of S. typhimurium phage type 208 from several Middle Eastern countries. The cultures carry a drug resistance plasmid of the FIme compatibility group, and at least two other plasmids which were detected and identified by gel electrophoresis. The studies supported and extended the genetic findings and provided information on the distribution of particular plasmids.
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A series of 2092 cultures of Salmonella typhimurium isolated from human, animal and other sources in 57 countries were differentiated into 204 phage types and 19 primary and 147 full biotypes. Different biotypes belonged to the same phage type and different phage types to the same biotype, so the combination of typing methods differentiated strains more finely than either method alone: 574 different ;phage type/biotypes' were distinguished in 1937 cultures belonging to the 204 recognized phage types.The combination of biotyping with phage-typing was valuable in studying the phylogeny and spread of epidemic strains by distinguishing clones of different biotype within the same phage type and by confirming the relationship between cultures isolated from widely dispersed clones and that between cultures isolated before and after a clone had undergone variation in phage type, biotype, colicin type or antibiotic-sensitivity pattern.A widespread outbreak of infection with S. typhimurium phage type 141 in Scotland comprised independent dissemination of three clones of different biotypes, 1f, 9f and 31bd. During its epidemic spread in cattle in Britain between 1962 and 1969, another strain underwent variations in phage type (type 44 to type 29), biotype (type 26a to types 26d, 26bd, 26dgi, 26dz and 26i) and antibiotic sensitivity. A group of 275 non-fimbriate, non-inositol-fermenting and non-rhamnose fermenting (FIRN) strains, particularly associated with avian infections and thought to be clonal in origin, contained 27 phage types and 22 full biotypes in the primary biotypes 29-32.
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Strains of Salmonella typhimurium of predominantly Middle Eastern origin, but distributed from England to India, were found to carry at least three types of resistance plasmid. The most important was initially identified as an F(I) plasmid by compatibility tests, but differs from the F factor on the one hand and the F(I) factors R162 and ColV on the other. The three groups of F(I) plasmids can be distinguished by their compatibility reactions with the MP10 plasmid of S. typhimurium (Smith, Humphreys, Grindley, Grindley & Anderson, 1973) and group H(1) factors: the F factor is unilaterally incompatible with group H(1) (Smith, Grindley, Humphreys & Anderson, 1973; Anderson, 1975b); the F(I) factors are compatible with MP10 and group H(1); and F(I)me factors are incompatible with MP10 but compatible with H(1). The majority of S. typhimurium cultures belonged to phage type 208; most of those that did not, belonged to types related to 208. Only a minority of their F(I)me plasmids were autotransferring. The remainder were mobilizable by F-like plasmids, and by group H(1) and H(2) factors, but not by the fi(-) I(1) factor Delta, or by plasmids of the I(2), B, P, W, N and com 7 groups. The compatibility reactions of the autotransferring F(I)me plasmids were identical with those of the non-transferring members of the group, and both were large, single-copy plasmids.The S. typhimurium strains of this series carried A or AK, and SSu resistance determinants: small, probably multicopy, non-transferring plasmids similar to those originally described in phage type 29 of S. typhimurium (Anderson & Lewis, 1965b).These S. typhimurium cultures probably represent a clone of wide geographical distribution. The accurate epidemiological study of such clonal outbreaks requires, in addition to phage typing, precise identification of the plasmids harboured by the epidemic strains, and may have to be carried to the molecular level.F(I)me plasmids were identified in other drug-resistant salmonellas, notably in a strain of S. wien which caused large outbreaks of mainly paediatric infection in Algeria, and also spread to Britain. An F(I)me plasmid was found in S. typhi phage type 44 from Algeria, in which the phage-restricting properties of the plasmid are responsible for the specificity of the type.
The phage-typing scheme of Callow (1959) has been extended. The original number of types was 34; this has now risen to 207. Tables are presented which show the provisional type designations and the definitive designations now being introduced.
An investigation of in vitro mutagenesis of plasmid DNA with hydroxylamine is described. The treated plasmid DNA was used to transform Escherichia coli K12. Mutants of the plasmid NTP3, which codes for resistance to ampicillin and sulphonamides, were isolated and characterised. They were classified according to the reduction in level of their beta-lactamase activity. Hydroxylamine-induced mutants of NTP14 were also isolated. This plasmid codes for ampicillin resistance, synthesis of colicin E1, and the EcoRI restriction and modification enzymes. One class of mutants is lethal to the host strain at temperatures above 33 degrees C, but carrier strains grow well at 28 degrees C. There is evidence that these mutants code for a temperature-sensitive EcoRI modification activity: the lethal effect probably results from the cleavage of the host-cell DNA by the restriction enzyme at non-permissive temperatures. The possible genetic uses of the mutant plasmids for the production of hybrid plasmids in the bacterial cell are discussed.
The properties of two plasmids coding for the CcoRI restriction and modification enzymes are described. Both plasmids are non auto-transferring (NTP) but can be mobilised by transfer factors. Strains carrying NTP13 produce colicin E1 and the EcoRI enzymes. This plasmid has a molecular weight of 6 X 10(6) daltons and is present as approximately 12 copies per chromosome. The second plasmid, NTP14, was detected after mobilisation of the EcoRI plasmid with the R factor RI-19. NTP14 codes for ampicillin resistance, synthesis of the EcoRI enzymes and colicin E1. The molecular weight of NTP14 is 10.7 X 10(6) daltons and there are about 14 copies per chromosome. DNA-DNA reassociation experiments were performed to determine the interrelationships of NTP13, NTP14, ColE1 and the R factor R1-19. NTP13 and NTP14 continue to replicate when cellular protein synthesis is inhibited by the addition of chloramphenicol.
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Polyethylene glycol quantitatively precipitates plasmid DNA of molecular weight 6-123-10-6, from cleared lysates of plasmid-carrying bacterial strains, After resuspension and density-gradient centrifugation of the precipitated DNA, it is unchanged in length and in transformation efficiency for Escherichia coli K12. Plasmid DNA can be easily prepared in large quantities by including a polyethylene glycol precipitation step in standard plasmid isolation procedures.
Transferable chloramphenicol resistance has become common in the typhoid bacillus in countries such as Mexico, India, Vietnam and Thailand. Situations such as this, and others analogous to it in many parts of the world, are the result of the long-term indiscriminate use of chloramphenicol and other antibiotics in the affected areas. They can be rectified only by more rational antibiotic usage.
The molecular length and DNA homology of R factors isolated from enterobacteria of human and animal origin have been examined. DNA from plasmids of the same compatibility group, whether of human or animal origin is indistinguishable, after allowance has been made for the regions coding for different antibiotic resistances. These results indicate that there is a common pool of R factors in man and ainmals.
A new, two-tier system for biotyping Salmonella typhimurium gives a finer and more reliable differentiation of strains than the Kristensen scheme and is capable of future extension by the addition of new types and new tests. Strains are allocated to a primary type (1-32) by their reactions in five primary tests with Bitter's xylose medium, meso-inositol, L-rhamnose, d-tartrate and m-tartrate. Subtypes are distinguished within the primary types by reactions in ten secondary tests, which include observations for flagella and type-1 (haemagglutinating) fimbriae. Full biotypes are designated by letters indicating the subtype reactions appended to the primary-type numbers. A series of 2030 strains of S. typhimurium collected from many different sources and countries during 53 years was classified into 19 of the 32 potential primary biotypes and into 144 full biotypes. Of the series, 14% (275) were non-fimbriate inositol-nonfermenting rhamnose-nonfermenting (FIRN) strain in primary biotypes 29-32. Most other strains were fimbriate and rhamnose fermenting. Observations on several series of cultures isolated from different human or animal sources in the same epidemic showed that the biotype characters of a strain were generally stable during its growth in the natural environment and in the unselective media used for isolation and storage. Most non-fermenting strains gave rise to fermenting mutants on prolonged incubation in the substrate-containing--and therefore selective--test medium, and false-positive results from this cause were avoided by making the definitive readings of tests after a short, carefully chosen period of incubation. A genealogical tree has been drawn to show how eighteen observed primary biotypes may have evolved from a presumed archetypal ancestor of biotype 1.
A controlled field trial was performed in Egypt to evaluate a whole cell typhoid vaccine prepared with a nonmotile mutant of S. typhi Ty2 (TNM1) devoid of flagellar antigen. This vaccine did not elicit an H antibody response, but significant Vi and O agglutinin responses were observed. There were 34 typhoid cases among 21 063 six- to seven-year-old children who received the TNM1 vaccine, and 44 cases among 21 017 children in the control group who received tetanus toxoid. These results suggest that TNM1 vaccine does not provide protection against typhoid fever, and that H antigen may be an important component of an effective vaccine.