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L K Ng

Publications and source records attributed to L K Ng.

At least 37 records · Page 2Linked to original sources

Evaluation of Etest for testing antimicrobial susceptibilities of Neisseria gonorrhoeae isolates with different growth media.

The MICs for 101 isolates of Neisseria gonorrhoeae obtained by Etest (AB-Biodisk, Solna, Sweden) and the agar dilution method on GC medium base supplemented with 1% Kellog's defined supplement (GCMB) were compared. The overall percent agreement (within 1 log2 dilution) between methods was greater than 97.9. The Pearson's correlation coefficients for penicillin, tetracycline, erythromycin, and ceftriaxone for the two methods were 0.98, 0.97, 0.93, and 0.93 (P = 0.001), respectively, for comparisons on GCMB. The overall percent agreement was lower when hemoglobin-supplemented GCMB was used. Etest is an attractive alternative to the agar dilution method for gonococcal antimicrobial susceptibility testing and should be further analyzed in multicenter studies.

Culture Media↗

Typing by serovar, antibiogram, plasmid content, riboprobing, and isoenzyme typing to determine whether Neisseria gonorrhoeae isolates requiring proline, citrulline, and uracil for growth are clonal.

Neisseria gonorrhoeae isolates requiring proline, citrulline, and uracil for growth (PCU-) have homogeneous phenotypes; most are plasmid-free, belong to few serovars, and are significantly associated with intermediate levels of susceptibility to penicillin, tetracycline, erythromycin, and cefoxitin. Because of their lack of variation by these criteria, molecular typing methods, ribotyping (restriction fragment length polymorphism [RFLP] of rRNA genes), and multilocus enzyme electrophoresis were explored as tools for further distinguishing PCU- isolates. By ribotyping, selected PCU- isolates could be separated into four groups on the basis of the hybridization patterns (RFLPs) of SmaI- and AvaII-digested DNA with probes containing rRNA sequences. Most of the isolates (18 of 23 isolates) belonged to a single RFLP (group I). One isolate each was in groups II and IV, and three isolates were in group III. All isolates except one, isolate NS791, had similar multilocus enzyme electrophoresis patterns. Strain NS791 was unusual in that it contained a variant cryptic plasmid with an insert in the 0.46-kb MspI-HinfI fragment of the 4.2-kb plasmid, it was the only isolate belonging to RFLP group IV, and it differed in its multilocus enzyme electrophoresis pattern, having different mobilities for glyceraldehyde phosphate dehydrogenase, phosphoglucose isomerase, 6-phosphogluconate dehydrogenase, and glutamate dehydrogenase. Serovars of PCU- isolates appeared to be more indicative of strain divergence than RFLP or isoenzyme typing. Multilocus enzyme electrophoresis indicated that PCU- isolates are clonal.

Bacterial Typing Techniques↗

Molecular fingerprinting of isolates of the genus Peptostreptococcus using rRNA genes from Escherichia coli and P. anaerobius.

Restriction fragment length polymorphisms (RFLPs) of rRNA genes were evaluated as a tool for intra- and interspecies differentiation of Peptostreptococcus isolates. RFLPs from a collection of 20 clinical isolates and five ATCC strains representing five Peptostreptococcus spp. (P. anaerobius, P. asaccharolyticus, P. magnus, P. micros and P. prevotii) were obtained by hybridization of Southern blots of HindIII- or EcoRI-digested genomic DNA with three probes: probe A, a 0.98 kb HindIII fragment with a partial 16S rRNA gene sequence from P. anaerobius ATCC 27337; probe B, cloned Escherichia coli rrnB operon in plasmid pKK3535; and probe C, E. coli 16S and 23S rRNA. The hybridization patterns varied, but all yielded RFLPs useful for both intra- and inter-species differentiation. RFLPs of P. asaccharolyticus clinical isolates were closely related to each other and differed significantly from those of the ATCC type strains. The profiles of P. prevotii differed from those of the other four species studied, and based on the HindIII- and EcoRI-generated RFLPs, the strains in this species are more heterogeneous than the other four species studied.

Cloning, Molecular↗

Plasmid analysis and antimicrobial susceptibilities of Peptostreptococcus species.

There are no published methods on plasmid isolation from Peptostreptococcus spp., therefore two methods of plasmid isolation from this genera were analysed: the boiling and alkaline-SDS methods. Plasmid DNA was not recovered by the boiling method, however, with the alkaline-SDS method, cryptic plasmid DNA was detected in two P. asaccharolyticus and one P. magnus strains. To achieve optimum lysis, Peptostreptococcus cells were treated with lysozyme (2 mg/ml) for 15 min. at 37 degrees C followed by proteinase K (0.2 mg/ml) for 1 h at 37 degrees C. In addition we report, the occurrence of clindamycin or metronidazole-resistant peptostreptococci, but these phenotypes were not correlated with plasmid carriage.

DNA, Bacterial↗

Strategies for molecular characterisation of methicillin- and gentamicin-resistant Staphylococcus aureus in a Canadian nosocomial outbreak.

Sixteen methicillin-resistant Staphylococcus aureus (MRSA) isolates, from a single nosocomial outbreak, were tested for molecular and phenotypic relationships. Two of the 16 outbreak strains were gentamicin resistant (Gmr) and the plasmids that they carried were characterised by reverse field electrophoresis, restriction endonuclease analysis and gene hybridisation. The gentamicin-resistant (Gmr) strains harboured two plasmids, a Gmr plasmid of 36.5 kb and a cryptic plasmid of 25.4 kb, whereas the other 14 isolates contained only the cryptic plasmid. Gentamicin resistance was encoded by a 2.5-kb HindIII fragment of the 32.8-kb plasmid and is similar to the 2.5-kb HindIII fragment also described for S. aureus Gmr plasmids from Australia and the USA. The Gmr plasmid was non-conjugal and was cured by ethidium bromide at a frequency of 4%. Two MRSA strains isolated subsequently from the same hospital were also Gmr and had identical plasmid and restriction endonuclease profiles to the two Gmr strains studied initially. Two other S. aureus isolates from the original carrier detected in this study and from his son were methicillin and gentamicin susceptible and had novel profiles. Since large plasmids show anomalous migration in agarose gels, more definitive analyses than simple plasmid identification should be considered when studying nosocomial outbreaks.

Cross Infection↗

The effects of cranial TENS on measures of autonomic somatic and cognitive activity.

Peripheral and central nervous system effects of the cranial application of sub-threshold transcutaneous electrical nerve stimulation (TENS), of sinusoid waveform passed between earlobe electrodes at an AC frequency of 100 Hertz, were investigated. In a single-blind study, each of thirty healthy volunteer subjects was administered one 30 minute treatment of either active TENS, placebo TENS or no treatment. Pretreatment to post-treatment changes in measures of autonomic activity (blood pressure, pulse rate, peripheral vasomotor activity), somatic activity (skeletal muscle tension), and anxiety were evaluated. Significant reductions in systolic blood pressure (p less than .05), diastolic blood pressure (p less than .01), pulse rate (p less than .05), peripheral vasomotor activity (marginally significant: p less than .07) and anxiety (p less than .05) were observed subsequent to active. TENS as compared to both placebo TENS and no TENS. No significant placebo TENS effect was observed. Possible mechanisms of action of this form of cranial TENS on the peripheral and central nervous system are discussed.

Adolescent↗

Characterization of freshly isolated Campylobacter coli strains and suitability of selective media for their growth.

Typical and atypical Campylobacter strains were isolated from the colons of cattle and swine by techniques that enabled the selective pressures of antibiotics to be avoided. Some cephalothin-susceptible strains and a strain with an indeterminate hippurate reaction were classified as Campylobacter coli by DNA homology testing. Tetracycline-resistant isolates were obtained from animals with no recorded exposure to antibiotics. A selection of 12 C. coli and 6 C. jejuni strains was used to determine the ability of fresh isolates to grow on a range of selective media. C. coli isolates were inhibited more than C. jejuni on selective media containing antibiotics. The least inhibitory media were Skirrow medium (M. B. Skirrow, Br. Med. J. 2:9-11, 1977) and the charcoal-based media developed by Hutchinson and Bolton (D. N. Hutchinson and F. J. Bolton, J. Clin. Pathol. 37:956-957, 1984) and Karmali et al. (M. A. Karmali, A. E. Simon, M. Roscoe, P. C. Fleming, S. S. Smith, and J. Lane, J. Clin. Microbiol. 23:456-459, 1986). The plasmid contents of the isolates did not appear to be related to their sensitivity to growth on selective (antibiotic-containing) media. The study indicates that selective media used to detect Campylobacter spp. could select against the isolation of C. coli.

Animals↗

Classification of Campylobacter strains using DNA probes.

DNA homology determinations were used to classify atypical strains of Campylobacter spp. and Campylobacter-like organisms. Using the slot-blot hybridization technique, atypical strains were classified as C. coli or C. jejuni. Some of the Campylobacter-like organisms isolated from animals were catalase-positive, but they did not hybridize with the DNA probes prepared from the total genomic DNA of C. coli, C. jejuni, C. laridis, C. fetus subsp. fetus and C. hyointestinalis. The taxonomic classification of these strains remains uncertain. The slot-blot hybridization technique was simplified by applying whole cells from a 24 h broth culture, instead of pure DNA to nitrocellulose membranes. The results of DNA homology from both methods were comparable, and the classification of these organisms using these methods was the same. Therefore, the dot-blot hybridization method is recommended as a more economical and time-saving method for use in the taxonomic classification of atypical Campylobacter spp.

Base Composition↗

Isolation and characterization of Campylobacter pyloridis from gastric biopsies.

Gastric biopsy specimens were examined microbiologically and histologically for the presence of Campylobacter pyloridis. Of 51 randomly selected patients, 22 (43%) were found to harbor C. pyloridis in the gastric mucosa. The histologic demonstration of spiral organisms observed by staining with hematoxylin and eosin correlated well with microbiologic isolation of the organisms. There was a strong association (95.5%) between C. pyloridis in the gastric mucosa and histologically defined gastritis. However, there was no obvious association between C. pyloridis and ulcers. All C. pyloridis strains isolated exhibited uniform biochemical characteristics and had almost identical protein profiles, which indicated that they belong to a relatively homogeneous group distinct from other Campylobacter species. All C. pyloridis isolates were uniformly susceptible to ampicillin, amoxicillin, cephalothin, gentamicin, kanamycin, tetracycline, coumermycin, ciprofloxacin, novobiocin, clorobiocin, and nitrofurantoin. They were moderately resistant to nalidixic acid.

Campylobacter↗

DNA probes for identification of tetracycline resistance genes in Campylobacter species isolated from swine and cattle.

Tetracycline-resistant strains of Campylobacter jejuni and Campylobacter coli from swine and cattle colons were isolated and characterized by hybridization with DNA probes. A probe consisting of the 1.8-kilobase (kb) HincII fragment from pUA466 was highly specific for the detection of tetracycline resistance (Tcr) in C. jejuni and C. coli. The 5-kb tetM DNA probe from Streptococcus agalactiae plasmid pJI3 which has homology with the 1.8-kb HincII fragment from pUA466 could also be used to detect Tcr Campylobacter strains. However, the tetM probe had a much lower sensitivity and required a lower stringency of hybridization. Therefore, the 1.8-kb HincII fragment appeared to be more appropriate for the classification of Tcr in Campylobacter spp. No homology was detected between the Tcr determinant from Campylobacter spp. and the tetL and tetN probes from Streptococcus spp. DNA homology was demonstrated between pUA649, a derivative of plasmid pUA466 which had lost most of the Tcr region, and Tcr plasmids from C. jejuni and C. coli isolated from animal and human sources. There was also homology between pUA649 and the chromosomes of C. jejuni and C. coli strains. In this study, all but one of the tetracycline-resistant C. coli and C. jejuni strains contained plasmids of approximately 50 kb which hybridized with the 1.8-kb HincII probe. In one C. coli strain (UA703), Tcr appeared to be chromosomally mediated.

Animals↗

Constant current sine wave transcutaneous nerve stimulation for the evaluation of peripheral neuropathy.

We have evaluated various forms of peripheral neuropathy with a new device which emits a constant sinusoid stimulus at varying frequencies to quantitate current perception thresholds (CPTs). In normal individuals, CPT measures increase with increasing stimulation frequency, are highest on the toe and lowest on the face. There is a significant effect of age and sex on threshold perception. In patients with neuropathy, there is a marked increase in thresholds on hands and feet, as well as a lesser but still significant increase of facial CPTs. Thresholds furthermore correlated with clinical severity in a group of patients with diabetic neuropathy. Facial thresholds were markedly elevated in the patients with moderate to severe neuropathy, suggesting that the device is sensitive to the systemic nature of peripheral neuropathy. The authors believe the device will be a useful tool in screening for sensory neurologic abnormalities.

Adult↗

New screening device for assessment of peripheral neuropathy.

Screening for the onset of toxic and entrapment neuropathy is a major concern in occupational medicine today. Current perception thresholds may be used as a measure of the integrity of the peripheral nervous system. A new transcutaneous nerve stimulator was used to evaluate current perception thresholds in 54 normal persons and 33 diabetic subjects. Current perception thresholds in the normal volunteers with no evidence of peripheral neuropathy were found to vary significantly with the frequency and location of the stimulation, as well as age. The test identified the diabetic peripheral neuropathy with an overall sensitivity of 94%. This new diagnostic technique is quick, simple to perform, noninvasive and nonaversive and provides a sensitive quantitative measure of sensory function. This diagnostic stimulator will be useful for screening occupationally related toxic and entrapment neuropathies in which sensory impairment is an early finding.

Adult↗

Transcutaneous nerve stimulation. Frequency and waveform specificity in humans.

The effects of transcutaneous electrical nerve stimulation (TNS) of a constant alternating current administered at various frequencies and waveforms to volunteer human subjects were investigated. The TNS was found to evoke noncutaneous subjective sensations in all the subjects. Only with a sinusoid waveform of TNS were distinct frequency ranges of the stimulation associated with specific noncutaneous subjective sensations. Our findings suggest that nervous tissue is capable of discriminating the waveform parameters of an electrical stimulus.

Adult↗

Susceptibility of Campylobacter species to nalidixic acid, enoxacin, and other DNA gyrase inhibitors.

Nalidixic acid-resistant mutants of Campylobacter jejuni and C. coli as well as "C. laridis" strains showed cross-resistance to another DNA gyrase subunit A inhibitor, enoxacin (MIC, 32 micrograms/ml), whereas C. fetus subsp. fetus, C. fetus subsp. venerealis, and "C. hyointestinalis" strains were all susceptible to enoxacin (MIC, less than or equal to 2 micrograms/ml). All Campylobacter species were resistant to novobiocin (MIC, 32 to 512 micrograms/ml), but most strains were susceptible to the other DNA gyrase subunit B inhibitors coumermycin A1 and clorobiocin.

Anti-Bacterial Agents↗

Estimation of Campylobacter spp. in broth culture by bioluminescence assay of ATP.

The luciferin-luciferase bioluminescence reaction was used to estimate cell numbers of Campylobacter jejuni and Campylobacter coli in broth cultures based on a linear relationship between cell numbers (in excess of 10(4) to 10(5] and ATP levels. The sensitivity was lower than that obtained with Escherichia coli. The calculated amount of intracellular ATP per cell of C. jejuni and C. coli ranged from 1.7 to 2.1 fg.

Adenosine Triphosphate↗

Comparison of basal media for culturing Campylobacter jejuni and Campylobacter coli.

Four strains of Campylobacter jejuni and four strains of Campylobacter coli were used to compare the quantitative growth of Campylobacter cells on blood agar base no. 2 (Oxoid), brucella agar (BBL Microbiology Systems and Difco Laboratories), campylobacter agar base (Difco), Columbia blood agar base (Difco and Oxoid), and Mueller-Hinton agar (Difco and Oxoid). Columbia blood agar base and blood agar base no. 2 were inhibitory to most of the strains tested, as evidenced by reduced (10- to 1,000-fold) colony counts compared with other basal media. One of the brucella agars was inhibitory to two of the C. coli strains. The inhibitory effect of these media could be eliminated by addition of FBP (0.05% each ferrous sulfate hydrate, sodium metabisulfite, and sodium pyruvate) or 7% defibrinated sheep blood. However, addition of FBP or blood to brucella agar, campylobacter agar base, or Mueller-Hinton agar did not significantly affect the count, indicating that supplements are not required in these media for growth of Campylobacter in pure culture.

Campylobacter↗

Inhibition of Campylobacter coli and Campylobacter jejuni by antibiotics used in selective growth media.

The ability of Campylobacter coli and Campylobacter jejuni to grow in the presence of antibiotics used in selective growth media was compared. MIC data for C. coli indicated that some strains were more susceptible to the antibiotics than were the C. jejuni strains tested. A reduction of greater than 1 log cycle in the numbers of cells growing on plates containing antibiotics was considered to be a marked level of inhibition. Only one of nine of the antibiotic combinations studied did not markedly inhibit most of the C. coli strains tested. Although one C. coli strain was not inhibited by any of the antibiotic combinations, the other six strains were inhibited for up to 7 log cycles. The addition of blood or growth supplements reduced but did not eliminate the inhibitory effect. The inhibition of laboratory strains of C. coli on media developed for the isolation of Campylobacter spp. indicates that the incidence of C. coli may be underestimated.

Anti-Bacterial Agents↗

Biochemical characteristics and identification of Enterobacteriaceae isolated from meats.

The isolation and identification of 2,220 Enterobacteriaceae from meats indicated that Escherichia coli biotype I, Enterobacter agglomerans, and Serratia liquefaciens were the principal types to be differentiated in meats. Citrobacter freundii, Klebsiella pneumoniae, Enterobacter cloacae, and Enterobacter hafniae were also commonly identified. Identification of isolates by the Encise II (Roche Diagnostics Inc., Nutley, N.J.) and Minitek (BBL Microbiology Systems, Cockeysville, Md.) coding systems gave similar results with only 255 (11.5%) discrepancies in identity, but both systems required large numbers of supplementary tests for identification of the isolates. Not only the distribution of Enterobacteriaceae types isolated from meats but also some of the biochemical reactions of the isolates differed from those of clinical isolates. The Minitek technique is recommended because of its versatility. However, with the addition of cellobiose and salicin disks and the inclusion of methyl red to the Minitek test and the use of the Voges-Proskauer test and gas production in EC medium at elevated temperature as standard tests, the identification of these Enterobacteriaceae from meats would be greatly facilitated. The inclusion of the motility test, for example, using nitrate motility agar, would also be of value to Enterobacteriaceae identification.

Carbohydrate Metabolism↗