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How long should patients with Salmonella typhi or Salmonella paratyphi be followed-up? A comparison of published guidelines.

Records were examined for 242 individuals infected with Salmonella typhi or S. paratyphi identified in Birmingham between 1981 and 1988, with a total of 335 person years of follow-up. Of these cases 77 and 78 per cent respectively were followed beyond the point at which surveillance would have ceased under guidelines published by the American Public Health Association and by the Public Health Laboratory Service for England and Wales. Under these two sets of guidelines only seven (3.8 per cent) and eight (4.3 per cent) cases respectively had subsequent positive faecal or urine cultures over a median of 335 and 295 days of additional follow-up. After 0, 1, 2, 3, 4 and 5 prior consecutive negative sets of cultures obtained at weekly intervals the likelihood of the next set of cultures being positive was 26, 9, 5, 2.2, 2.4 and 0 per cent respectively. Only 38 (1.7 per cent) of 2184 follow-up urine cultures were positive; these results did not influence duration of follow-up. Only 26 (2.6 per cent) of 1002 contacts were infected; the yields of the first, second and third sets of cultures were 1.5, 0.6 and 0.5 per cent respectively.

Adolescent↗

Epidemiological analysis of Salmonella typhi-murium infections on the basis of laboratory methods. I. Distribution of phage types and biotypes of Salmonella typhi-murium isolated in Hungary in the period 1960 to 1981.

Phage and biochemical types were determined of 34 937 Salmonella typhi-murium cultures including 31 708 human strains, 2732 animal strains and 497 strains isolated from water. Phage type 4, not typable strains (nt) and phage type 2b were predominant among the strains of human and animal origin, and nt, 4 and 2b among the strains isolated from water. The most frequent phage types and the nt strains were subdivided by biotyping and additional phages. The incidence of S. typhi-murium var. copenhagen strains was 12.4%, they belonged mainly to phage types 2b and nt. The number of S. typhi-murium isolates of human origin showed a 2-4 year periodical fluctuation between 1960 and 1981. A connection was found between the incidence of the predominant phage types (4, nt, 2b) and the periodical changes in the total number of isolates. Phage type 4, which predominated among the strains of human and animal origin till 1976, was ousted gradually by nt ones. In the period when the change in predominance was observed the number of epidemics decreased and the number of sporadic cases increased. The change in the frequency of phage types took place at the same time when the frequency of phage types changed among the strains isolated from cattle and meat-products (4----nt). The increased number of sporadic cases after 1976 refers to infections from cattle and not from poultry.

Animals↗

Vaccination of calves against Salmonella dublin with aromatic-dependent Salmonella typhimurium.

Ten Holstein calves were divided into 2 groups. Five calves served as nonvaccinated controls, and 5 calves were vaccinated IM at 2 and 3 weeks of age with 10(9) aromatic-dependent (aro-) Salmonella typhimurium strain SL1479 containing O antigens 1, 4, 12. Serious adverse reactions to vaccination were not observed in the calves. Mean maximum rectal temperature increase in the vaccinated calves was 1.5 C. One calf had diarrhea and depressed appetite for 1 day after vaccination. At 5 weeks of age, all calves were challenge exposed orally with 1.5 X 10(11) virulent S dublin strain SL1367 (O antigens 9,12). After challenge-exposure inoculum was given, 1 of 5 vaccinated calves died and 4 of the 5 nonvaccinated calves died (P less than 0.05). Thus, some cross serotype protection against S dublin was induced by parenteral vaccination of calves with aro- S typhimurium strain SL1479, although protection was not complete.

Animals↗

Proteins from Salmonella R-mutants mediating protection against Salmonella typhimurium infection in mice I. Preparation of proteins free from lipopolysaccharide using various chromatographic methods.

Different chromatographic methods are described to remove traces of LPS from proteins extracted from the surface of several R-mutants of Salmonella. The protein preparations obtained by these methods were found to contain less than 0.001% of beta-hydroxymyristic acid as determined by gas liquid chromatography. This corresponds to less than 0.003 to 0.006% of R-form-LPS (depending on the chemotype of the mutant) and less than 0.014% of S-form-LPS. From some protein mixtures LPS could be removed by repeated gel filtration on a column of Sepharose CL-6B. Recycling gel filtration proved to be more effective considering the high recovery of protein in much shorter time. Ion exchange chromatography on DEAE Sepharose CL-6B was also useful in some cases. Thus, using one or the other method LPS free protein preparations could be obtained with extracts obtained from 6 out of 7 bacterial strains.

Animals↗

Interactions Between Macrophages of Guinea Pigs and Salmonellae II. Phagocytosis of Salmonella typhimurium by Macrophages of Normal Guinea Pigs.

Peritoneal macrophages from normal guinea pigs were allowed to phagocytize Salmonella typhimurium in vitro. The extent of phagocytosis was determined by quantitative viable counts of the bacteria released after lysis of the phagocytes with sodium deoxycholate. It was shown that the avirulent strain RIA of salmonellae was more susceptible to ingestion by macrophages than the virulent strain SR-11. The presence of immune serum in the phagocytic mixture greatly enhanced the extent with which strain SR-11 was phagocytized. Also, the virulent bacteria recovered from infected mice exhibited a greater resistance to phagocytosis than those maintained in artificial media.

Journal Article↗

A study of drug resistance among Salmonella typhi and Salmonella paratyphi A in an endemic area, 1977-79.

Tests for antibiotic resistance were carried out on 198 strains of Salmonella typhi and S. paratyphi A isolated from cases of enteric fevers. Their minimal inhibitory concentrations for streptomycin, chloramphenicol, ampicillin, furazolidine and co-trimoxazole were estimated by plate dilution technique. Four strains of S. typhi and one strain of S. paratyphi A were found to show multiple resistance with a set pattern of resistance to chloramphenicol, streptomycin, sulphonamide, tetracycline and spectinomycin. All the five strains carried R-factors. Three of the resistant S. typhi belonged to Phage type 'O' and one was in Phage type 'A'. The single resistant S. paratyphi A belonged to Phage type '2'.

Ampicillin↗

Identification of contemporary plasmid virulence genes in ancestral isolates of Salmonella enteritidis and Salmonella typhimurium.

Six epidemiologically distinct ancestral strains of Salmonella enteritidis and 5 of S. typhimurium from the pre-antibiotic era were examined for plasmid content, and for presence of plasmid genes implicated in mouse-virulence. Five sizes of plasmid were detected in S. enteritidis varying from 1 to 60 MDa. Two sizes of plasmid were found in S. typhimurium, 28 and 60 MDa. Plasmids of the same size were not common to both serovars. The HindIII restriction patterns of 3 of the ancestral S. enteritidis plasmids were identical to the modern 38 MDa plasmid, while all contained identical bands of 3.5, 2.7 and 1.9 kb. All the 60-MDa S. typhimurium plasmids, ancestral and contemporary, had an identical restriction pattern. Three different sized S. enteritidis plasmids and one size S. typhimurium plasmid contained a 3.5-kb DNA fragment carrying the virulence locus VirA. The VirB virulence locus was located on a 2.7-kb DNA fragment in S. enteritidis and on a 2.5-kb fragment in S. typhimurium. Both loci were precisely conserved between the ancestral strains and the modern representatives of both serovars.

DNA Fingerprinting↗

Effect of different purine auxotrophic mutations on mouse-virulence of a Vi-positive strain of Salmonella dublin and of two strains of Salmonella typhimurium.

Ten cfu of a Vi-positive strain of Salmonella dublin, given i.p. to BALB/c mice, caused death. Of transductional derivatives with nutritional requirements expected to reduce virulence, those with complete blocks at aroA (therefore aromatic-dependent) or at purA (therefore adenine-dependent) were non-virulent; no deaths from i.p. inocula of 10(6) or 10(7) cfu, respectively. Transductants with identified blocks at purF, purG, purC or in the purJHD operon, all with purine requirement satisfied by hypoxanthine or any other purine, were of reduced virulence, with some deaths from 10(4) cfu or, for most of the derivatives, from only 10(2) cfu. Blocks at guaA or guaB, causing guanine requirement, likewise resulted only in reduced virulence, with some deaths from inocula of 10(2). Transductional derivatives of two mouse-virulent (LD50, i.p., less than 25 cfu) strains of S. typhimurium were similarly tested, with generally similar results, in that purA and purB (adenine-requiring) transductants were non-virulent, no deaths from 2.5 x 10(7) cfu, and those with purF, purG or purC defects (hypoxanthine-responding) retained partial virulence. However, the guaA and guaB (guanine-requiring) derivatives were of much reduced virulence; all mice survived 2.5 x 10(5) cfu but only one survived 2.5 x 10(7) cfu. In both the S. dublin and the S. typhimurium tests autopsy cultures showed that fatal infections resulted from multiplication of the auxotrophic strain, not from that of non-exacting revertants.

Animals↗

Vaccination of chickens with a Salmonella enteritidis aroA live oral Salmonella vaccine.

A mouse-virulent strain of Salmonella enteritidis, Se795 (LD50 less than 10 organisms for mice), was non-virulent for 12-day-old chickens given 10(6) cfu intravenously; the organisms were cleared from liver and spleen by day 14 as measured by direct plating and by day 21 by enrichment. An Se795aroA mutant, CU58, was also cleared from liver and spleen by day 14 after intravenous inoculation of 10(7) cfu. Day-old chicks vaccinated orally with either one dose of 10(9) CU58 at 1 day of age, 10(7) at 1 and 14 days, or 10(5) at 1 and 7 days followed by 10(9) at 14 and 21 days of age, were challenged orally with a nalidixic acid resistant variant of the virulent phage type 4 S. enteritidis strain 109. All vaccinated groups showed a reduction in faecal shedding of the challenge. Chickens given four doses of CU58 showed a significant reduction of cfu in liver, spleen and faeces following intravenous challenge with virulent strain 109. Intramuscular vaccination with 10(9) cfu of Aro strain CU58 at 1 day of age gave no protection against oral challenge with virulent strain 109. Serum antibody production to LPS (ELISA) was minimal in all vaccinated birds. The results indicate that oral vaccination with Aro- S. enteritidis can confer protection to day old chicks against virulent S. enteritidis.

Animals↗

A Salmonella-based, propionate-inducible, expression system for Salmonella enterica.

The expression and regulatory properties of a propionate-regulated overexpression system (Salmonella enterica prpBCDE promoter (P(prpB)) and positive regulator (prpR)) were evaluated in wild-type S. enterica serovar Typhimurium TR6583 and prpB(-) or prpD(-) versions of this strain and compared with the arabinose-regulated T7 expression system. The wild-type strain showed low expression in the absence of propionate and high expression in the presence of propionate under all growth conditions. In 96-well plates and culture tubes, the wild-type strain exhibited a long delay before full induction; the time delay was significantly shorter in shake flasks. The prpD(-) strain exhibited low expression in the presence of glucose, highly regulatable expression over a wide range of propionate concentrations, and, in contrast to the wild-type strain, fast induction to full expression under all growth conditions. In contrast, the prpB(-) strain showed very high background expression in both culture tubes and shake flasks.

Arabinose↗

Proteins from Salmonella R-mutants mediating protection against Salmonella typhimurium infection in mice. II. Protection tests performed with proteins free from lipopolysaccharide.

Lipopolysaccharide-free proteins obtained from R-mutants of Salmonella typhimurium, S. minnesota and S. dublin as well as those from an S-form of S. typhimurium mediated protection in mice against experimental infection with S. typhimurium. The protection was measured by LD50 and is statistically significant. The level of protection of all the preparations was similar. Non-bacterial proteins had no effect under similar experimental conditions. Protection afforded by the purified proteins was lower than that of the corresponding complex crude extract; supplementation of proteins with lipopolysaccharide and phospholipids in model membrane vesicles enhanced their potency. However, lipopolysaccharides or phospholipids alone were not able to increase the efficacy of purified proteins. Some other fractions obtained by gel filtration of the crude extract also afford protection.

Animals↗

Survival of Salmonella east bourne and Salmonella typhimurium in chocolate.

Experiments were carried out to assess the reduction rate of two salmonella strains (S. eastbourne and S. typhimurium) in chocolate bars. After artificial contamination of chocolate, after 'conching', with about 10(6) S. eastbourne/g. this organism was still recovered after 9 months storage. The strain of S. typhimurium was less resistant. Both serotypes died off more rapidly in bitter chocolate than in milk chocolate. After contamination with a smaller dose (about 10(3)/g.) with these two serotypes, similar differences were observed.

Cacao↗

Utilization of d-tartaric acid by Salmonella paratyphi B and Salmonella java: comparison of anaerobic plate test, lead acetate test and turbidity test.

d-Tartrate dehydrase of Salmonella java is an oxygen-sensitive enzyme active in cultures incubated under the poorly aerated conditions of static culture but not in fully aerated shaken cultures nor on plates incubated aerobically. On plates of d-tartrate minimal agar incubated anaerobically the enzyme or the degradation products of d-tartrate are exported from d-tartrate-positive cells and are available to d-tartrate-negative bacteria. This may give misleading growth results when d-tartrate-positive and d-tartrate-negative strains are tested for growth on the same plate of d-tartrate minimal agar. The lead-acetate test terminated at 24 h, the 24 h turbidity test and the ability to grow on d-tartrate minimal agar within 48 h differentiated 53 S. paratyphi B strains that were negative in each of the three tests from 76 S. java that were positive in each of the tests. An intermediate group of eight strains utilized d-tartrate in Difco bacto-peptone water to give a positive lead acetate reaction at 2 days, were stimulated to a varying degree by d-tartrate in Oxoid peptone water within the same period of incubation and grew poorly on d-tartrate minimal agar. These latter strains may be deficient in a permease controlling uptake of d-tartrate or export of d-tartrate dehydrase. Inability to utilize d-tartrate is unlikely to be the single character accountable for the reputed enhanced pathogenicity of S. paratyphi B when compared with S. java. Indications for the existence of an enzyme, complementary to and mutually exclusive with d-tartrate dehydrase, that has a positive correlation with pathogenicity are discussed.

Anaerobiosis↗

The geographical distribution of Salmonella typhi and Salmonella paratyphi A and B phage types during the period 1 January 1970 to 31 December 1973. A report of the International Federation for Enteric Phage-Typing (IFEPT).

The frequency distribution is presented of the Vi-phage types of Salmonella typhi and of the phage types of S. paratyphi A and S. paratyphi B, examined by the National Centres represented in the IFEPT from 1970 to 1974. The most common types encountered in this international study were A and E1 (= E 1 a), as has been previously observed. There was, however, a wide range of types, many of which are characteristic of particular geographical zones. The occurrence of such types out of their habitual context indicates their 'exotic' origin, providing a useful clue to their probable source. Plasmid-mediated chloramphenicol resistance appeared in epidemic typhoid fever for the first time during this period. It occurred in Mexico, India, Vietnam and Thailand, and has assumed important proportions. All the plasmids concerned belonged to compatibility group H 1. The study of this phenomenon continues, and the findings will appear in future reports.

Bacteriophage Typing↗

Presentation of Salmonella antigens by peritoneal cells of normal and Salmonella-infected mice.

A comparison of the ability of normal peritoneal cells (PC) and those harvested from mice 1-3 days after intraperitoneal immunization with live Salmonella enteritidis 11RX (11RX) to present antigen to 11RX-primed T cells was made using formalin-killed 11RX and a soluble 11RX antigen extract as antigens. Unfractionated PC and the adherent and non-adherent PC populations were analysed separately and the effects of the lysosomal function-impairing drug chloroquine and the fixative paraformaldehyde, used before or after antigen-pulsing, were also determined. The results presented indicate that immunization with live 11RX did not induce any detectable modulation of APC function which could account for the ability of live 11RX to induce cell-mediated immune responses involving Lyt 2+ T cells.

Animals↗

A fluorescent amplified fragment length polymorphism study of Salmonella enterica serovar Sofia, the major Salmonella serovar isolated from chickens in Australia.

Fluorescent amplified fragment length polymorphism (FAFLP) analysis was performed on 68 isolates of Salmonella enterica subsp. salamae serovar Sofia (S. Sofia). Fifty eight isolates were obtained over a period of approximately 15 years from a range of human, chicken industry and environmental sources throughout Australia. A further ten isolates were identified from human and poultry sources in Israel from 1972 to 1987. Analysis of FAFLP profiles for fragments between 50 to 500 base pairs in length indicated distinct clusters of isolates. All but seven isolates clustered into four groups of >90% similarity and all isolates displayed at least 70% similarity with each other. No cluster could be attributed to a particular geographical, temporal or source-of-isolation origin. It is concluded that S. Sofia is genetically variable with certain clones persisting over time but no group appears unique to Australia.

Animals↗

Bacteriophages mediating somatic antigenic conversion in Salmonella cholerae-suis: their isolation from sewage and other Salmonella serotypes possessing the somatic 6 antigen.

Bacteriophages which mediate the conversion of the O somatic antigen of Salmonella cholerae-suis from the 6(2)7 to the 6(1)7 phenotype have been isolated from two strains of S. newport and one of S. muenchen, and also from sewage collected from two areas where there have been no reports of S. cholerae-suis infection for several years. The phages differed from each other by cross-resistance tests.

Antigens, Bacterial↗

Salmonella typhimurium DT 193: differentiation of an epidemic phage type by antibiogram, plasmid profile, plasmid fingerprint and salmonella plasmid virulence (spv) gene probe.

Of over 2000 isolates of Salmonella typhimurium DT 193 from humans examined in the 2 year period 1991-92, 93% were antibiotic-resistant with the most common R-types being ASSuT (38%) and T (29%). Fourteen plasmid profiles were identified in DT 193 R-type ASSuT with the majority of isolates being characterized by a single plasmid of 80 MDa (pDEP 34) which in addition to coding for ASSuT, also hybridized with a spv gene probe prepared from the 50 MDa Salm. dublin serovar-specific plasmid. On the basis of restriction fragment length polymorphisms, two variant lines of pDEP 34-like plasmids were identified and a third line which had lost the genes coding for resistance to ampicillin, streptomycin and sulphonamides, was recognized. Although 18 plasmid profile types were identified in DT 193 R-type T, all isolates carried a high mol. wt plasmid which coded for tetracycline resistance only. Further discrimination was achieved on the basis of hybridization of tetracycline resistance plasmids with the spv gene probe and restriction enzyme fingerprinting. These results demonstrate that Salm. typhimurium DT 193 can be rapidly subdivided by antibiogram and that further subdivision can be achieved on the basis of plasmid profile, plasmid fingerprint and hybridization with a spv gene probe.

Animals↗