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Biomedical subjects

W M Johnson

Publications and source records attributed to W M Johnson.

At least 37 records · Page 2Linked to original sources
PubMed1997

Screening for Salmonella with a murine monoclonal antibody M105 detects both Felix O1 bacteriophage sensitive and resistant Salmonella strains.

Ten Felix O1 (FO1) bacteriophage sensitive Salmonella strains as well as their phage resistant derivates together with 39 strains of FO1-resistant Salmonella were tested for their reactivities with a murine monoclonal antibody, M105, by indirect whole cell and competitive ELISA. All FO1 phage sensitive and 48 of the 49 FO1-resistant Salmonella strains were found to react with M105. The single Salmonella strain not reacting with M105 was a FO1 resistant derivative selected by exposing the sensitive parent strain to the phage. This M105-negative and FO1-resistant strain was also found to be a rough mutant without O-antigens and possibly lacks the terminal LPS core sugars which form the M105 reactive epitope.

Antibodies, Bacterial↗
PubMed1997

Salmonella isolated from humans, animals and other sources in Canada, 1983-92.

A total of 89760 human and 22551 non-human isolates of salmonella were serotyped in Canada during the period 1983-92. There were 2180 reported outbreaks associated with 10065 cases during the 10-year period. The most common salmonella serovars isolated from human and non-human sources were S. typhimurium and S. hadar. The third and fourth most common serovars from human sources were S. enteritidis and S. heidelberg, respectively, and from non-human sources they were S. heidelberg and S. infantis. The number of S. typhimurium isolations from human and non-human sources showed a downward trend over the 10-year period. A total of 222 outbreaks of S. typhimurium associated with 1622 cases occurred. The S. hadar isolations from human and non-human sources reached a peak during the years 1987-90 and declined thereafter. The number of human isolates of S. enteritidis increased until 1985 and fluctuated at a level of 8.3-12.8% of all human isolates thereafter. Seventy-three outbreaks of S. enteritidis infection associated with 568 cases occurred. More than 50% of the S. enteritidis infections in humans were caused by phage type (PT) 8. During the review period, infections caused by PT4 were less common and were almost exclusively found in people who had travelled abroad. The annual isolation rates of S. heidelberg from human and non-human sources increased steadily during the period. Bacteriophage typing of serovars from outbreaks showed that contaminated food products of poultry and bovine origin were common sources of human infection. Salmonella typhi was identified as the cause of 43 small outbreaks affecting 116 persons.

Animals↗
PubMed1997

Human salmonellosis associated with exotic pets.

During the period from 1994 to 1996, an increase in the number of laboratory-confirmed cases of human salmonellosis associated with exposure to exotic pets including iguanas, pet turtles, sugar gliders, and hedgehogs was observed in Canada. Pet turtle-associated salmonellosis was recognized as a serious public health problem in the 1960s and 1970s, and in February 1975 legislation banning the importation of turtles into Canada was enacted by Agriculture Canada. Reptile-associated salmonellosis is once again being recognized as a resurgent disease. From 1993 to 1995, there were more than 20,000 laboratory-confirmed human cases of salmonellosis in Canada. The major source of Salmonella infection is food; however, an estimated 3 to 5% of all cases of salmonellosis in humans are associated with exposure to exotic pets. Among the isolates from these patients with salmonellosis, a variety of Salmonella serotypes were also associated with exotic pets and included the following: S. java, S. stanley, S. poona, S. jangwani, S. tilene, S. litchfield, S. manhattan, S. pomona, S. miami, S. rubislaw, S. marina subsp. IV, and S. wassenaar subsp. IV.

Animals↗
PubMed1997

Genetic identification of Mycobacterium bovis BCG by restriction fragment length polymorphism analysis of the direct-repeat region.

Restriction fragment length polymorphism (RFLP) analysis was performed on the direct repeat (DR) regions of 14 strains of Mycobacterium bovis BCG. With AluI-digested DNA, BCG Japanese, Russian, and Mexican had differing RFLP patterns but 11 strains, including Pasteur, Glaxo, and Tice, had an identical pattern not detected in over 60 strains of the M. tuberculosis complex. DR analysis can aid in confirming the identification of clinical BCG isolates.

DNA, Bacterial↗
PubMed1997

Analysis of genetic polymorphism in the phospholipase region of Mycobacterium tuberculosis.

mtp40 was originally identified as a short genomic region that was found in strains of Mycobacterium tuberculosis but not in Mycobacterium bovis. Subsequent studies have revealed that the sequence is part of the mpcA gene, which encodes a phospholipase C. To investigate further the distribution of the mtp40 sequence, we analyzed strains of the M. tuberculosis complex by PCR and were able to amplify the mtp40 sequence in 90 of 94 strains of M. tuberculosis and in 2 strains of Mycobacterium microti but not in M. bovis or M. bovis BCG. Based on this, we developed a dot blot assay using genomic DNA which allows M. bovis to be distinguished from the majority of M. tuberculosis strains. We also probed Southern blots of 140 clinical isolates of M. tuberculosis to determine the frequency of strains lacking mtp40. This revealed an unexpected polymorphism in the phospholipase region. Two fragments were detected in 57% of samples. The expected fragment of 0.75 kbp corresponds to the region of mpcA containing mtp40. A 2.1-kbp fragment was observed to belong to a recently discovered second phospholipase gene, mpcB. In addition, some strains appeared to lack both genes, while others showed only the presence of mpcA. A few strains had additional bands, suggesting the existence of other homologs to the two phospholipase genes. We also detected the insertion of IS6110 in the mpcA coding region of one strain. The absence of these genes in some clinical isolates raises questions about their function during infection and in the development of tuberculosis disease in humans.

Genes, Bacterial↗
PubMed1996

Immunochemical characterization of a haemagglutinating antigen of Arcobacter spp.

The Arcobacter haemagglutinin has been identified by Western immunoblot to be an immunogenic protein of about 20 kDa. The haemagglutinating activity is sensitive to proteolytic enzyme digestion and heat treatment of 80 degrees C and above. The Arcobacter haemagglutinin is possibly a lectin-like molecule binding to erythrocytes via a glycan receptor containing D-galactose as part of its structure.

Animals↗
PubMed1996

Environmental gasoline-utilizing isolates and clinical isolates of Pseudomonas aeruginosa are taxonomically indistinguishable by chemotaxonomic and molecular techniques.

A total of 42 Pseudomonas aeruginosa strains was isolated previously from clinical sources (27 strains) and from a gasoline-contaminated aquifer (15 strains). Selected strains were subjected to taxonomic tests involving chemical and molecular biological techniques, including membrane fatty acid analysis, phage-sensitivity, growth temperature range, presence of plasmids, and PCR-amplification and sequencing of a species-specific 16S-23S rDNA internal transcribed spacer region. The clinical and environmental isolates formed a coherent taxonomic group with few distinguishing characteristics. Of the phenotypes observed, a consistent difference was the ability of the aquifer strains to utilize gasoline supplied in the gas phase as sole carbon source and, conversely, the inability of the clinical strains to do so. Fourteen of the 15 environmental strains possessed similar-sized cryptic plasmids. The clinical isolates either lacked detectable plasmids or contained plasmids of a different size. The observation that the clinical and environmental isolates of P. aeruginosa were taxonomically indistinguishable is discussed in terms of its relevance to environmental-regulatory guidelines because P. aeruginosa, a known opportunistic pathogen, is a prime candidate for use in bioremediation processes involving deliberate release of this organism to the environment.

Bacteriophages↗
PubMed1996

A selective medium for the isolation of Arcobacter from meats.

A method, including enrichment in Arcobacter Selective Broth (ASB) and isolation on semisolid Arcobacter Selective Medium (ASM) under aerobic conditions at 24 degrees C, is described for the isolation of Arcobacter from retail meat products. Selective agents used in ASB and ASM were cefoperazone, trimethoprim, piperacillin and cycloheximide. Arcobacters were isolated from 53 (24.1%) of 220 poultry meat products and also, at lower incidence from samples of beef and pork. The isolates were identified as A. butzleri or A. butzleri-like and belonged to a wide variety of serotypes and biotypes.

Animals↗
PubMed1996

Characterization of cytotoxic, hemolytic Aeromonas caviae clinical isolates and their identification by determining presence of a unique hemolysin gene.

Aeromonas caviae has recently been recognized as an important enteropathogen and its hemolysin is purported to be one of the virulence factors, In this study, a total of 80 clinical isolates of Aeromonas spp. were investigated by PCR with synthetic oligonucleotides targeting a cloned hemolysin-encoding sequence from an A. caviae isolate of clinical origin. Of the 35 clinical A. caviae isolates tested, only 6 contained the target sequence.

Aeromonas↗
PubMed1996

Evaluation of ribotyping as epidemiologic tool for typing Escherichia coli serogroup O157 isolates.

A total of 121 representative Escherichia coli O157:H7 and O157:NM (nonmotile) isolates were characterized by ribotype, phage type, verotoxin genotype, and genomic fingerprints generated by pulsed-field gel electrophoresis. Ribotyping was not able to discriminate between O157:H7 isolates, and phage typing and pulsed-field gel electrophoresis were the most valuable and discriminatory techniques.

Bacterial Typing Techniques↗
PubMed1996

Identification of IS1356, a new insertion sequence, and its association with IS402 in epidemic strains of Burkholderia cepacia infecting cystic fibrosis patients.

Burkholderia cepacia is now recognized as an important opportunistic pathogen in cystic fibrosis (CF) and other compromised patients. Epidemicity among CF patients has been attributed to at least one particularly infectious strain (strain ET12), and both genetic evidence and anecdotal evidence suggest that this strain, currently endemic in Ontario, and those causing an epidemic in the United Kingdom, are indeed the same. Our study was conducted to determine whether there was any association between the presence of various insertion sequence (IS) elements, the cable pilin subunit gene (cblA), electrophoretic type (ET), and ribotype (RT) in a collection of 97 clinical and 2 environmental isolates of B. cepacia. No apparent linkage was found for IS elements IS401, IS402, IS406, IS407, and IS408 with ET or RT. The cblA target, said to be a marker for high infectivity, was detected in 100% (38 of 38) of strains of B. cepacia ET12 and in a single strain of ET13 that differed in a single enzyme allele. A new IS, IS1356, identified during the investigation, was present in 71.7% of all isolates, and 50.7% of these isolates harbored IS1356 as a hybrid IS element inserted into IS402. IS1356 is 1,353 bp in length, and when it is inserted into IS402 it results in a 10-bp duplication at the site of insertion. IS1356 contains one major open reading frame of 1,260 bp coding for a putative transposase which has significant homology to ISRm3 in Rhizobium meliloti (59%) and to an undesignated IS element in Corynebacterium diphtheriae (49%). The IS402-IS1356 element was found exclusively in the epidemic strains from Ontario and the United Kingdom, being detected in 94.7% (36 of 38 isolates) of B. cepacia ET12 isolates. Of the two ET12 isolates found to be devoid of the IS402-IS1356 element, both contained IS1356 unassociated with IS402, one was temporally unrelated to the epidemic, and the other was from a CF patient in a geographic area remote from Ontario and the United Kingdom. It is evident that the IS402-IS1356 hybrid element, the cblA pilin subunit gene, and the allelic suite represented by multilocus enzyme electrophoretic type ET12 may provide useful markers for the epidemic, highly transmissible transatlantic strain isolated in Ontario and the United Kingdom.

Amino Acid Sequence↗
PubMed1995

Identification of epidemiologic markers for Neisseria meningitidis using difference analysis.

The feasibility of identifying epidemiologic markers based solely on the identification of DNA fragments present in outbreak-associated isolates was investigated using Neisseria meningitidis (Nm) as a model system. The clonal structure of Nm has been well characterized using multilocus electrophoresis. In Canada, electrophoretic types ET1, ET5, ET9 and ET21 are being displaced from the natural population by type ET15, and the latter type is associated with an increased prevalence of serogroup C meningococcal disease. Difference analysis, which uses subtractive hybridization and polymerase chain reaction (PCR) amplification, was employed to identify amplifiable DNA fragments (amplicons) that differ between the ET15 and the ET1, ET5, ET9 and ET21 genomes. 14 amplicons were cloned which were further characterized by Southern blot analysis to identify six amplicons that represent fragments either unique to or highly polymorphic in the ET15 genome. Oligodeoxyribonucleotide primer pairs were designed for each of the six amplicons, and PCR amplification was used to determine their prevalence across a panel of 167 Nm isolates representative of other serogroups and ETs. Among group C isolates only two of the six amplicons, designated as A and G, were effective in discriminating ET15 from non-ET15 isolates. Amplicon A detects a deletion in the dhps gene which effectively differentiates sulfonamide-sensitive and -resistant serogroup C isolates. The frequency of amplicon A and G detection in the other serogroups and ETs was too great to facilitate their direct use as diagnostic markers for the differentiation of virulent Nm isolates.

Bacterial Outer Membrane Proteins↗
PubMed1995

Oligonucleotide primers designed to differentiate pathogenic pseudomonads on the basis of the sequencing of genes coding for 16S-23S rRNA internal transcribed spacers.

Universal primers targeting conserved sequences flanking the 3' end of the 16S and the 5' end of the 23S rRNA genes (rDNAs) were used to amplify the 16S-23S rDNA internal transcribed spacers (ITS) from eight species of pseudomonads which have been associated with human infections. Amplicons from reference strains of Pseudomonas aeruginosa, Pseudomonas cepacia, Pseudomonas gladioli, Pseudomonas mallei, Pseudomonas mendocina, Pseudomonas pickettii, Pseudomonas pseudomallei, and Xanthomonas maltophilia were cloned from each species, and sequence analysis revealed a total of 19 distinct ITS regions, each defining a unique sequevar with ITS sizes ranging from 394 (P. cepacia) to 641 (P. pseudomallei) bp. Five distinct ITS sequevars in P. cepacia, four in P. mendocina, three in P. aeruginosa, two each in P. gladioli and P. pseudomallei, and one each in P. mallei, P. pickettii, and X. maltophilia were identified. With the exception of one P. cepacia ITS, all ITS regions contained potential tRNA sequences for isoleucine and/or alanine. On the basis of these ITS sequence data, species-specific oligonucleotide primers were designed to differentiate P. aeruginosa, P. cepacia, and P. pickettii. The specificities of these primers were investigated by testing 220 clinical isolates, including 101 strains of P. aeruginosa, 103 strains of P. cepacia, and 16 strains of P. pickettii, in addition to 24 American Type Culture Collection (ATCC) Pseudomonas strains. The results showed that single primer pairs directed at particular ITSs were capable of specifically identifying the ATCC reference strains and all of the clinical isolates of P. aeruginosa and P. pickettii, but this was not the case with several ITS-based primer pairs tested for P. cepacia. This pathogen, on the other hand, could be specifically identified by primer pairs directed against the 23S rDNA.

Base Sequence↗
PubMed1994

Comparison by extended ribotyping of Pseudomonas cepacia isolated from cystic fibrosis patients with acute and chronic infections.

Multiple isolates of Pseudomonas cepacia, from two cystic fibrosis (CF) patients who were chronically infected and two others who suffered acute fatal lung infections, were examined by multilocus enzyme electrophoresis and four-enzyme ribotyping. The strains isolated from the fatalities belonged to a clone, electropherotype 12 (ET12) that is endemic in the Ontario patients' province of origin. ET12 strains have also been isolated from outbreaks in CF patients in the United Kingdom, where they are considered to be strains of high virulence and transmissibility and epidemiologically related to Ontario strains. Four-enzyme ribotyping (EcoRI, Xho, PstI, and ClaI) established the close genetic relationship of the Ontario ET12 isolates and those from the United Kingdom, particularly an isolate from Manchester. In addition, four enzyme ribotypes of the sequential isolates taken during life and at autopsy of the ET12 clone were highly variable in comparison with the stability of the ribotypes of clone ET16 isolated sequentially from living chronic carriers. This extreme ribotype variability may be indicative of a highly virulent strain and poor prognosis. Isolates from our chronically infected CF patients belonged to a different clone, ET16, and it is also endemic in its region, 1000 miles east of ET12, in Nova Scotia. In both endemic circumstances, person-to-person transmission was easily demonstrated by four-enzyme ribotyping. The ET12 clone was found to be transmitted among summer campers and during a nosocomial outbreak, whereas an E16 strain was found to infect a sibling of a chronically infected patient; both infections were of the same ribotype.

Acute Disease↗