Isolation of CDC group M-5 and Staphylococcus intermedius from infected dog bites.
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Biomedical subjects
Publications and source records attributed to B Holmes.
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Twenty-one type or other reference strains, each representing a different Campylobacter, Helicobacter, or Arcobacter taxon, and a reference strain of Staphylococcus aureus were used to assess the reproducibility of nine enzyme detection tests used in the identification of campylobacters. For five of the tests (alkaline phosphatase, DNase, and H2S production, indoxyl acetate hydrolysis, and nitrate reduction), more than one procedure was employed to determine the most suitable method. Alkaline phosphatase test results were better defined and more reproducible if read after 1 h of incubation. Detection of DNase was fully reproducible with each method (except with Helicobacter pylori), but reactions were generally weaker than those of other DNase-producing organisms. Both procedures for determining H2S production were irreproducible for the same strains. The reproducibility of indoxyl acetate hydrolysis was improved by using disks impregnated with 25 microliters of substrate. Reduction of nitrate was best determined by Cook's plate method. Results for the other tests examined (catalase, oxidase, and urease production and hippurate hydrolysis) were both pertinent and fully reproducible for all strains.
This study compared the effect of varied training frequencies on the development of isometric lumbar extension torque (strength) over 12- and 20-week training periods. Fifty-six subjects were randomly assigned to training once every other week (training group 1, n = 10), once per week (training group 2, n = 12), twice per week (training group 3, n = 12), or three times per week (training group 4, n = 7) or to a nontraining control group (n = 15). Training consisted of one set of 8 to 12 variable-resistance lumbar extensions to volitional muscular fatigue. Prior to and following 12 and 20 weeks of training, subjects were given a test that evaluated isolated isometric lumbar extension torque in a seated position at seven positions (angles) through a 72-degree range of motion. The control group showed no change in isometric torque. All training groups showed significant increases in lumbar extension torque at 12 and 20 weeks of training, whereas no significant differences were found among the groups with respect to the magnitude of torque gained. Pooled training showed a significant time x angle interaction at 12 weeks and a continuing trend at 20 weeks, indicating that the shape of the isometric torque-angle curve changed as a result of training. This effect was due to greater increases in isometric torque at the fully extended position than at the fully flexed position at 12 weeks (92% versus 16%, respectively) and at 20 weeks (123% versus 17%, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)
Seventy-five strains of Proteus mirabilis (mainly of human origin) were characterized by one-dimensional SDS-PAGE of cellular proteins. The strains came from various countries; 24 were from urine, 17 from faeces (including one from a dog and one from a monkey), 12 from the respiratory tract (including nine from sputum), four from a cerebral abscess, 12 from miscellaneous sources and six from unknown sources. The protein patterns, which contained 45 to 50 discrete bands, were highly reproducible. The patterns of the 75 P. mirabilis cultures plus those of the type strains of six Proteus and Providencia species were used as the basis for two numerical analyses. In the first, which included all the protein bands, the P. mirabilis cultures formed nine clusters at the 85% S level. In the second analysis, in which the principal protein bands (in the 34.0-44.6 kDa range) were excluded, 74 of the 75 cultures of P. mirabilis formed a single cluster at the 77% S level distinct from the six Proteus and Providencia reference strains. The P. mirabilis strain which failed to cluster with the others had a background band pattern typical of the species in the lower molecular weight region but appeared to be less typical in the heavier bands. It is concluded that high resolution PAGE combined with computerized analysis of protein patterns provides the basis for typing clinical strains of P. mirabilis. Reference strains of each of the nine PAGE types identified are available from NCTC for inclusion in future studies.
The type strains of six Campylobacter species or subspecies were examined in eight tests used for the identification of such organisms. False-positive results were obtained in certain tests (growth on 1% bile, brilliant green, selenite, trimethylamine-N-oxide, 2,3,5-triphenyl tetrazolium chloride, and minimal media) when an inoculum yielding 10(7) to 10(8) CFU/ml was used. Each tolerance test was examined with blood, nutrient, and brucella agars as basal media. The type of basal medium used could also affect the test outcome. With the inoculum standardized to a density yielding approximately 10(6) CFU/ml, reproducible and pertinent results were obtained, provided an appropriate basal medium was used. However, 95% confidence limits for viable counts done on these basal media indicated that blood agar may yield more consistent results than the other agars examined.
Twenty type or other reference strains, each representing a different Campylobacter, Helicobacter, or Arcobacter taxon, were used to assess the reproducibility of 25 phenotypic tests that are used in the identification of such organisms. Twenty-two of the tests depended on growth inhibition, and each of these tolerance tests was performed by using three different basal media. Although the overall reproducibility of the tests with each basal medium exceeded 89%, the proportion of strains that were able to grow in a reproducible manner on the basal media varied from 100% for blood agar and 50% for nutrient agar to 5% for brucella agar. In general, test reproducibility was highest with the basal medium that supported the most luxuriant growth. For the majority of tests, the basal medium which gave the optimum reproducibility could be determined.
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Following isolation of a multi-antibiotic-resistant pseudomonad from a newborn infant on admission to the Special Care Baby Unit and further isolation of apparently the same organism from two additional infants, a full investigation was instigated in an attempt to discover the source of the organism. This revealed a further 13 infants apparently colonized with the same organism. Repeated screening of the infants with a commercial sterile swab/transport medium failed to isolate the organism. Examination of bottles of the in-house transport medium, which had been stored under a sink, produced further isolates of the same organism. Water splashed from the sink was suspected as the ultimate source of contamination. Biochemical characterization showed that P. pickettii and at least one other Pseudomonas species were involved. The epidemiological, clinical and economic implications of the 'outbreak' are discussed together with the ultimate financial implications for investigation of such incidents.
Twenty-four cultures comprising 20 clinical isolates of 'Klebsiella aerogenes' from two hospitals, a reference strain of 'K. aerogenes' and the type strains of three other Klebsiella species, were characterized by one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of whole-cell proteins. The protein patterns were highly reproducible and were used as the basis of a numerical analysis which divided the clinical isolates into 12 protein types. Comparison with established typing methods indicated that the level of discrimination of SDS-PAGE was similar to that achieved with conventional typing methods but the strains were grouped differently. Protein typing subdivided five serotype K3 isolates that could also be distinguished by phage typing. Conversely, three strains of protein type 11 were clearly distinguishable by both serotyping and phage typing. We conclude that high-resolution SDS-PAGE of proteins provides an effective adjunct to other methods for typing isolates of 'K. aerogenes'.
Twenty-five cultures comprising 18 clinical isolates of Serratia marcescens from two hospitals, the type strain of S. marcescens, two reference strains of S. marinorubra, the type or a reference strain of three other Serratia species and a reference strain of undetermined species, were characterized by one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of whole-cell proteins. The protein patterns were highly reproducible and were used as the basis of a numerical analysis which divided the clinical isolates into eight protein types. Comparison with O-serotyping indicated that the level of discrimination by SDS-PAGE was similar. As with O-serotyping, a secondary scheme, such as phage typing, is necessary to differentiate strains of the same protein type. We conclude that high-resolution SDS-PAGE of proteins provides an effective adjunct to other methods for typing isolates of S. marcescens.
Forty-two cultures of pseudomonas comprising 28 clinical isolates from a pseudo-outbreak on a Special-Care Baby Unit and 14 reference strains, including 9 type strains, of various Pseudomonas species, were characterized by one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of whole-cell proteins. The protein patterns were highly reproducible and were used as the basis for a numerical analysis which divided the strains into 9 phenons. Two of the 28 clinical isolates were identified by biochemical tests as P. pickettii and their identification was confirmed by SDS-PAGE as they fell in the same phenon as the type strain of the species. The remaining 26 isolates, which could not be identified on phenotypic tests, fell in the same phenon as three reference strains of 'P. thomasii'. The protein patterns provided the first clear evidence that P. pickettii and 'P. thomasii' were separate taxa and that the 'outbreak' was polymicrobial in origin, in line with the probable aqueous source of contamination. We conclude that high-resolution SDS-PAGE of proteins provides an effective method of identifying and differentiating pseudomonads, especially where this cannot be done adequately using conventional biochemical tests.
Variation amongst Achromobacter-like strains was examined by DNA restriction endonuclease digestion and rDNA gene patterns generated using a non-radioactive probe. Chromosomal DNA was extracted from 12 cultures representing Achromobacter groups B, E and F, all from human blood cultures. DNA fingerprinting using EcoRI, Hae III or HindIII sub-divided the strains in a similar manner to that obtained by their protein patterns. The HaeIII patterns, with their small number of bands, were the easiest to interpret. The EcoRI patterns included a species-species triplet of bands but minor band patterns allowed further differentiation. The Achromobacter group F strains comprised a separate taxon and were distinct from the group B and E strains by all techniques examined. The study demonstrates that, in addition to total DNA digest analysis, rDNA gene restriction patterns provide a simple but discriminatory electrophoretic method for distinguishing within Achromobacter groups B and E.
Sixty-five strains of Morganella morganii (mainly of human origin) were characterized by one-dimensional SDS-PAGE of cellular proteins. The strains came from various countries; 13 were from stools (including one from a toucan), 13 from wounds, 11 from urine, five from blood (including one from a snake), five from the respiratory tract (four sputum, one lung), 12 from miscellaneous sources and six from unknown sources. The protein patterns, which contained 45 to 50 discrete bands, were highly reproducible. The patterns of 67 M. morganii cultures plus those of the type strains of seven Proteus and Providencia species were used as the basis for two numerical analyses. In the first, which included all the protein bands, the M. morganii strains formed 21 clusters at the 91% S level. In the second analysis, in which the principal protein bands (in the 31.6-43.2 kDa range) were excluded, the 67 M. morganii cultures formed a single cluster at the 80% S level distinct from the seven Proteus and Providencia reference strains. We conclude that high resolution PAGE combined with computerized analysis of protein patterns provides the basis for typing clinical strains of M. morganii. Reference strains of each of the 21 PAGE types identified are available from NCTC for inclusion in future studies.
Thirty-seven strains of Group EF-4 bacteria (from various countries) were characterized by one-dimensional SDS-PAGE of cellular proteins. They comprised 21 from dog-bite wounds of humans, three from cat-bite wounds of humans and five from human limb wounds which may have been inflicted by dogs or cats; there was also one each from a pet monkey, a tiger lung (fatal), a dog tonsil, a mouse, a cat liver, a wallaby mandible, a human vagina and one from a human limb wound which was apparently not inflicted by an animal. The protein patterns, which contained 45 to 50 discrete bands, were highly reproducible and were used as the basis for three numerical analyses. In the first, in which the principal protein bands (in the 34.8 to 41.3 kD range) were excluded, the 37 Group EF-4 strains formed, at the 62% S level, two major clusters corresponding to strains producing a dihydrolase for arginine and those not doing so. In the second analysis, which included all the protein bands and which was performed only on the 22 arginine-positive strains, two phenons formed (one of which could be further divided into two sub-phenons) at the 56% S level. The third analysis, also based on all the protein bands, divided the 15 arginine-negative strains into three clusters at the 56% S level. We conclude that high resolution PAGE combined with computerised analysis of protein patterns correlates exactly with the separation of Group EF-4 into two biovars (also with the distinction of the biovars on the basis of G + C content). Reference strains of each of the PAGE types identified are available from the NCTC for inclusion in future studies.
Thirty-two clinical strains representing 'Achromobacter' groups B, E and F were characterized by one-dimensional SDS-PAGE of cellular proteins. All the strains were isolated from blood samples from hospital patients in the United Kingdom. The protein patterns, which contained 40 to 45 discrete bands, were highly reproducible and were used as the basis for a numerical analysis which included all the protein bands. The 32 'Achromobacter' strains formed two clusters at the 77% S level. The first, phenon 1, included the 28 group B and the two group E strains and the second, phenon 2, contained the two strains of group F. The strains in each phenon were characterized by a clearly distinct pattern of protein bands. Phenon 1 could be further divided at the 87% S level into three subphenons which correlated with differences in the principal bands found between 40.0 and 42.5 kD. Strains of group E clustered with group B strains from which they could not be distinguished by protein patterns. We conclude that high resolution PAGE combined with computerized analysis of protein patterns provides a useful method for the classification of this group of bacteria. Reference strains of each of the PAGE types identified are available from NCTC for inclusion in future studies.
Eighty-six strains of Providencia stuartii (mainly of human origin) were characterized by one-dimensional SDS-PAGE of cellular proteins. The strains came from various countries; 52 were from urine, 11 from wounds, five from blood (one of these also from urine), four from ear infections, two each from faeces and sputum, one from 'alimentation' and nine from unknown sources. The protein patterns, which contained 45 to 50 discrete bands, were highly reproducible. The patterns of 46 Prov. stuartii strains (selected to represent the full range of protein pattern diversity) plus those of the type strains of the four other Providencia species were used as the basis for two numerical analyses. In the first, which included all the protein bands, the Prov. stuartii strains formed 13 clusters at the 88% S level. In the second analysis, in which the principal protein bands (in the 33.8-40.7 kDa range) were excluded, 45 of the 46 Prov. stuartii strains formed a single cluster at the 82% S level, whilst the four Providencia reference strains remained unclustered. The 40 strains of Prov. stuartii not included in the cluster analysis were assigned to a protein type by calculating their similarity with the strains in the database used for the cluster analysis. We conclude that high resolution PAGE combined with computerized analysis of protein patterns provides the basis for typing clinical strains of Prov. stuartii. Reference strains of each of the 13 PAGE types identified are available from NCTC for inclusion in future studies.
Fifty-eight males and 34 females with nongonococcal urethritis and/or cervicitis were treated to compare the efficacy and safety of 7-day regimens of oral ofloxacin 300 mg twice daily and doxycycline hyclate 100 mg twice daily. Forty-seven patients were randomized to receive ofloxacin and 45 patients to receive doxycycline. The microbiologic response rate was 97% (32/33) for both ofloxacin and doxycycline; the combined microbiologic and clinical cure rates were 98% for both treatment groups (ofloxacin 46/47, doxycycline 44/45). Ofloxacin was as effective as doxycycline in the treatment of chlamydial infections (96% vs. 100%). In patients with Ureaplasma urealyticum, the initial response was complete with either drug, but recurrence of infection was observed with both treatment groups (1 of 4 patients in the ofloxacin group and 2 of 11 patients in the doxycycline group). In the treatment of mixed Chlamydia trachomatis and U. urealyticum infections, all 5 patients treated with ofloxacin and 3 of 4 patients treated with doxycycline were cured. In symptomatic patients whose initial cultures were negative, clinical cures were complete with both drugs, but Ureaplasma was isolated at 3 or more weeks post-treatment in 2 patients treated with ofloxacin. In a study of single-dose ofloxacin treatment of uncomplicated gonorrhea, Neisseria gonorrhoeae was eradicated in all subjects, but C. trachomatis was not reliably eradicated. Both drugs were well tolerated with only minimal adverse effects reported in either treatment group. A multiple-dose regimen of ofloxacin appears to be a highly effective and well-tolerated alternative to doxycycline in nongonococcal sexually transmitted disease.