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O Olsvik

Publications and source records attributed to O Olsvik.

At least 37 records · Page 2Linked to original sources
PubMed1998

Virulence properties of Escherichia coli isolated from Ethiopian patients with acute or persistent diarrhoea.

Escherichia coli strains isolated from faecal specimens of 108 Ethiopian patients with acute watery diarrhoea (n = 30), acute bloody (n = 9), and persistent (n = 25) diarrhoea, and from 44 patients who recently had recovered from diarrhoea were analyzed for the presence of virulence factors using DNA probes, and for adhesion to HeLa cells. Eighty-two patients were under five years of age. Enterotoxigenic E.coli (ETEC) were most frequently isolated (63 patients, 58%). Eighteen of the ETEC strains also hybridized with probes for EPEC adherence factor (EAF) and Enteroaggregative (EAgg) adherence. Enteroaggregative E. coli (EAggEC) were more frequently isolated than EAF positive E.coli, and more frequently from patients with persistent diarrhoea (10/25) than from patients with acute diarrhoea (11/39). In total, 103 of the patients harboured faecal E. coli which hybridized with one or more of the virulence probes. Haemagglutination of one or more erythrocyte species was expressed by 65/70 strains. Using monoclonal antibodies to Colonization Factor Antigen I and Coli Surface antigens 1-5, only 18/66 strains were found to produce one or more of these adhesions and no more than 15 of 43 ETEC strains were agglutinated by the antisera to these adhesins. Forty-nine strains adhered to HeLa cells in autoaggregative (23 strains), localized (17 strains) or diffuse (9 strains) pattern. The study shows that E.coli strains carrying genes for the different virulence factors are prevalent in Ethiopia. Testing for the presence of these virulence factors, as well as for putative colonization factor antigens, should be included in epidemiological studies in this area.

Acute Disease↗
PubMed1997

Evolutionary relationships among members of the genus Chlamydia based on 16S ribosomal DNA analysis.

Nucleotide sequences from strains of the four species currently in the genus Chlamydia, C. pecorum, C. pneumoniae, C. psittaci, and C. trachomatis were investigated. In vitro-amplified RNA genes of the ribosomal small subunit from 30 strains of C. pneumoniae and C. pecorum were subjected to solid-phase DNA sequencing of both strands. The human isolates of C. pneumoniae differed in only one position in the 16S rRNA gene, indicating genetic homogeneity among these strains. Interestingly, horse isolate N16 of C. pneumoniae was found to be closely related to the human isolates of this species, with a 98.9% nucleotide similarity between their 16S rRNA sequences. The type strain and koala isolates of C. pecorum were also found to be very similar to each other, possessing two different 16S rRNA sequences with only one-nucleotide difference. Furthermore, the C. pecorum strains truncated the 16S rRNA molecule by one nucleotide compared to the molecules of the other chlamydial species. This truncation was found to result in loss of a unilaterally bulged nucleotide, an attribute present in all other eubacteria. The phylogenetic structure of the genus Chlamydia was determined by analysis of 16S rRNA sequences. All phylogenetic trees revealed a distinct line of descent of the family Chlamydiaceae built of two main clusters which we denote the C. pneumoniae cluster and the C. psittaci cluster. The clusters were verified by bootstrap analysis of the trees and signature nucleotide analysis. The former cluster contained the human isolates of C. pneumoniae and equine strain N16. The latter cluster consisted of C. psittaci, C. pecorum, and C. trachomatis. The members of the C. pneumoniae cluster showed tight clustering and strain N16 is likely to be a subspecies of C. pneumoniae since these strains also share some antigenic cross-reactivity and clustering of major outer membrane protein gene sequences. C. psittaci and strain N16 branched early out of the respective cluster, and interestingly, their inclusion bodies do not stain with iodine. Furthermore, they also share less reliable features like normal elementary body morphology and plasmid content. Therefore, the branching order presented here is very likely a true reflection of evolution, with strain N16 of the species C. pneumoniae and C. psittaci forming early branches of their respective cluster and with C. trachomatis being the more recently evolved species within the genus Chlamydia.

Animals↗
PubMed1996

Isolation of cmr, a novel Escherichia coli chloramphenicol resistance gene encoding a putative efflux pump.

A novel gene designated cmr, which mapped to 18.8 min of the Escherichia coli K-12 genome, was shown to mediate resistance to chloramphenicol when it was expressed from a multicopy vector. The accumulation of chloramphenicol was significantly less in cells overexpressing cmr than in control cells harboring the vector without insert. After the addition of a proton motive force blocker, the level of accumulation of chloramphenicol in the resistant cells rapidly approached the levels found in sensitive cells carrying only the chromosomal cmr. Northern (RNA) blot analyses revealed that the cmr gene is expressed as a 1.3-kb transcript. This size corresponds very well with a predicted size of 1,293 nucleotides (nt) based on the mapping of the transcription initiation site to a G residue 24 nt upstream of the start codon and the presence of a putative rho-independent terminator sequence ending 36 nt downstream of the 1,233-nt open reading frame encoding the putative Cmr protein. The 411-residue-long derived amino acid sequence contains 12 putative transmembrane segments and displays significant sequence similarities to several known drug resistance protein sequences of the major facilitator family. We provide evidence strongly suggesting that the resistance mediated by Cmr involves active exclusion of chloramphenicol.

Amino Acid Sequence↗
PubMed1995

Probes and polymerase chain reaction for detection of food-borne bacterial pathogens.

DNA-hybridization and the polymerase chain reaction (PCR) are techniques commonly used to detect pathogenic bacteria. In this paper, the use of these techniques for detection of Salmonella, E. coli, V. cholerae, non-O1 Vibrio, Yersinia enterocolitica, Campylobacter, Listeria monocytogenes, Staphylococcus aureus, Bacillus cereus, Clostridium perfringens, and C. botulinum is reviewed with emphasis on application in food microbiology. In food control, DNA-techniques have most often been used in a 'culture confirmation' fashion, i.e. bacteria are enriched and sometimes even purified by traditional culture procedures and thereafter identified by the use of DNA-based methods. The most desirable approach is, however, to detect organisms directly in the food, but major problems remain to be solved before this can be routinely performed.

Bacteria↗
PubMed1995

A transferable multiple drug resistance plasmid from Vibrio cholerae O1.

Ten multiple antimicrobial-resistant isolates of Vibrio cholerae O1 isolated from patients in Uganda were characterized, and the transferability of resistance to bacteria of diverse origins was investigated. The isolates were toxigenic and belonged to biotype E1 Tor, serotype Ogawa, and ribotype 8, and possessed a 130-MDa plasmid of incompatibility group 6-C. This plasmid, designated pRVC1, was shown to confer resistance to trimethoprim (mediated by a dhfrI gene), sulfonamides (a suII gene), tetracycline [a tet(C) gene], chloramphenicol (a catI gene), ampicillin (a beta-lactamase gene other than blaTEM or blaSHV), and streptomycin. pRVC1 proved to be transmissible at frequencies between 1 x 10(-1) and 5 x 10(-6) transconjugants per recipient to a variety of bacterial pathogens, including those of humans, animals, and fish. Most efficient transfer was observed from V. cholerae to strains of Shigella flexneri, Escherichia coli, Vibrio parahaemolyticus, and three Aeromonas species. The present in vitro study suggests that pRVC1 may spread from V. cholerae to other bacteria pathogenic to man, animals, and fish in natural environments.

Animals↗
PubMed1995

Molecular subtyping of toxigenic Vibrio cholerae O139 causing epidemic cholera in India and Bangladesh, 1992-1993.

Since October 1992, > 150,000 cases of cholera have been reported from India and Bangladesh; the great majority of Vibrio cholerae isolates belong to the newly established serogroup O139. To better understand the interaction of genetic and epidemiologic factors responsible for their sudden appearance and rapid spread, representative toxigenic V. cholerae O139 isolates were molecularly characterized and compared with a set of toxigenic V. cholerae O1 and non-O1/non-O139 strains. DNA sequences of the cholera toxin B subunit gene and multilocus enzyme electrophoresis markers of V. cholerae O139 strains were identical to those of V. cholerae O1 isolates of the seventh pandemic. Two distinct ribotypes and four pulsed-field gel electrophoretic patterns were observed for O139 strains. V. cholerae O139 strains were very similar to V. cholerae O1 strains of the seventh pandemic but clearly different from the toxigenic V. cholerae strains of serogroups other than O1 and O139.

Bacterial Typing Techniques↗
PubMed1994

Molecular subtyping of Vibrio cholerae O1 strains recently isolated from patient, food and environmental samples in Spain.

Nineteen Vibrio cholerae O1 strains isolated in Spain from patient, food and environmental samples in the period 1990-1992 were characterized by detection of cholera toxin by enzyme immunoassay, detection of cholera toxin gene by polymerase chain reaction, and by biotyping, ribotyping and pulsed-field gel electrophoresis. Ten isolates were toxigenic and were further characterized by multilocus enzyme electrophoresis. Molecular subtyping methods allowed precise differentiation between isolates, indicating their geographic origin. Isolates associated with the ongoing seventh pandemic were distinguishable from those associated with the present Latin American epidemic. All isolates from the environment and seafood were nontoxigenic, and were genetically different and more diverse than toxigenic isolates. The data suggest that a focus of endemic cholera does not exist in Spain, and that the analyzed nontoxigenic Vibrio cholerae O1 isolates from imported seafood were not a threat to public health.

Animals↗
PubMed1994

Magnetic separation techniques in diagnostic microbiology.

The principles of magnetic separation aided by antibodies or other specific binding molecules have been used for isolation of specific viable whole organisms, antigens, or nucleic acids. Whereas growth on selective media may be helpful in isolation of a certain bacterial species, immunomagnetic separation (IMS) technology can isolate strains possessing specific and characteristic surface antigens. Further separation, cultivation, and identification of the isolate can be performed by traditional biochemical, immunologic, or molecular methods. PCR can be used for amplification and identification of genes of diagnostic importance for a target organism. The combination of IMS and PCR reduces the assay time to several hours while increasing both specificity and sensitivity. Use of streptavidin-coated magnetic beads for separation of amplified DNA fragments, containing both biotin and a signal molecule, has allowed for the conversion of the traditional PCR into an easy-to-read microtiter plate format. The bead-bound PCR amplicons can also easily be sequenced in an automated DNA sequencer. The latter technique makes it possible to obtain sequence data of 300 to 600 bases from 20 to 30 strains, starting with clinical samples, within 12 to 24 h. Sequence data can be used for both diagnostic and epidemiologic purposes. IMS has been demonstrated to be a useful method in diagnostic microbiology. Most recent publications describe IMS as a method for enhancing the specificity and sensitivity of other detection systems, such as PCR, and providing considerable savings in time compared with traditional diagnostic systems. The relevance to clinical diagnosis has, however, not yet been fully established for all of these new test principles. In the case of PCR, for example, the presence of specific DNA in a food sample does not demonstrate the presence of a live organism capable of inducing a disease. However, all tests offering increased sensitivity and specificity of detection, combined with reduced time of analysis, have to be seriously evaluated.

Animals↗
PubMed1993

Detection of streptococcal pyrogenic exotoxin genes by a nested polymerase chain reaction.

Severe invasive disease associated with group A Streptococcus (GAS) has recently increased in frequency. Isolates of GAS from normally sterile sites were examined for the streptococcal pyrogenic exotoxin genes spe A, spe B and spe C to determine if they play a role in this disease. Four primers for each gene were used in a nested polymerase chain reaction (PCR) configuration. The first PCR generated fragments of 818, 1106, and 801 bp, respectively, for the extotoxin genes. The second PCR generated fragments of 500, 912 and 654 bp for the spe A, spe B and spe C genes using the fragments from the first PCR as template. Of 62 strains tested, 35 (56%) contained the spe A gene, and 17 (27%) contained the spe C gene. All GAS strains studied, regardless of disease association, contained the spe B gene. These data corroborate accumulating evidence that the genes encoding pyrogenic exotoxin types B and C are not associated with severe invasive streptococcal illness including streptococcal toxic shock-like syndrome. This PCR-based gene detection system has clinical and epidemiologic applications because of its ease of performance, non-isotope labelling, high specificity and sensitivity, and lack of requirement for purified DNA.

Bacterial Proteins↗
PubMed1993

The molecular epidemiology of cholera in Latin America.

To explain the sudden appearance and rapid spread of cholera in Latin America in January 1991, molecular techniques were used to define Vibrio cholerae O1 isolates from around the world. Restriction fragment length polymorphisms of rRNA and ctxA genes, DNA sequence of cholera toxin B subunit gene ctxB, and multilocus enzyme electrophoresis data were used to characterize 197 isolates. Worldwide, there are at least four distinct toxigenic El Tor V. cholerae O1 clones: the seventh pandemic (Eastern Hemisphere), US Gulf Coast, Australian, and Latin American. Nontoxigenic V. cholerae O1 previously isolated in Brazil, Mexico, and Peru are unlike current toxigenic isolates. The Latin American clone probably represents an extension of the seventh pandemic into the Western Hemisphere, while the US Gulf Coast clone most likely evolved separately. These data will be useful in monitoring the spread of cholera, determining the origin of outbreaks in both hemispheres, and implicating specific vehicles of transmission.

Alleles↗
PubMed1993

Use of automated sequencing of polymerase chain reaction-generated amplicons to identify three types of cholera toxin subunit B in Vibrio cholerae O1 strains.

Cholera toxin is the principal factor causing the profuse intestinal fluid secretion that is characteristic of cholera. The DNA sequences of the cholera toxin subunit B structural genes from 45 Vibrio cholerae O1 strains isolated in 29 countries over a period of 70 years were determined by automated DNA sequencing of polymerase chain reaction-generated amplicons. Three types of cholera toxin B subunit gene (ctxB) were identified. Genotype 1 was found in strains of classical biotype worldwide and El Tor biotype strains associated with the U.S. Gulf Coast, genotype 2 was found in El Tor biotype strains from Australia, and genotype 3 was found in El Tor biotype strains from the seventh pandemic and the recent Latin American epidemic. All base changes correspond to an amino acid substitution in the B subunit of the cholera toxin. Heterogenicity in the B subunit could have implications for vaccine development and diagnostic tests for cholera toxin and antitoxin. We conclude that this technology provides timely and potentially useful epidemiological information.

Africa↗
PubMed1993

Epidemiologic application of a standardized ribotype scheme for Vibrio cholerae O1.

A standardized scheme of 27 different BglI ribotypes and subtypes of Vibrio cholerae O1 strains is proposed on the basis of data from 214 human and environmental strains isolated in 35 countries and 14 U.S. states over the past 60 years. The ribotype patterns obtained are reproducible and stable over time. Seven different but very similar ribotypes (1a to 1g) were observed among 16 strains of the classical biotype. Twenty ribotypes and subtypes were identified among 198 V. cholerae O1 strains of the El Tor biotype. Six different patterns were found among the strains causing the current seventh pandemic. Strains of ribotype 8 originated only in central African countries, while those of ribotype 3 originated mainly in Asia and the Pacific Islands. The most widely distributed strains were those of ribotype 6, which was subdivided into three very similar but still distinguishable subtypes. The present Latin American epidemic is caused by strains of ribotype 5. Strains of this ribotype were isolated from several other geographic locations but can be differentiated from the Latin American strains by other molecular methods. Strains associated with two documented environmental reservoirs exhibited three distinct ribotype patterns; those isolated from patients who ate food from the U.S. Gulf waters were all of ribotype 2, while the strains related to the northeast Australian rivers were of ribotypes 9 and 10. Nontoxigenic V. cholerae O1 strains originating in Latin America and the U.S. Gulf Coast did not form a specific cluster of ribotypes. Ribotyping in combination with other well-defined methods can assist in epidemiologic investigations, helping to trace the movement of strains and to identify their geographic origins.

Animals↗
PubMed1993

Acquired tetracycline resistance genes in nosocomial Salmonella typhimurium infection in a Kenyan hospital.

Tetracyclines have been among the most widely used antibiotics worldwide. Plasmid-mediated tetracycline resistance among hospital strains of bacteria has continued to rise and of major concern has been the transfer of resistance to pathogenic organisms. Bacteraemia due to hospital acquired S. typhimurium has been a major cause of morbidity at Kenyatta National Hospital (KNH), hence the need to study drug susceptibility pattern of this organism. This study also characterized the tetracycline resistance genes using oligonucleotide probes. Ninety seven S. typhimurium strains isolated from patients at KNH were used. Agar dilution method was used to determine minimum inhibitory concentration (MIC). Plasmids were isolated from each strain and the different plasmid profiles were grouped by their molecular weights into 6 patterns. Out of 97, 87 (88%) strains were resistant. MIC ranged from 1 microgram/ml to 128 micrograms/ml. Genes encoding for tetracycline resistance were located on plasmids of molecular weights 65 MDa, 5.2 or both. Plasmid-encoded antimicrobial resistance is likely to spread to other pathogenic organisms, reduce our ability to treat the infection and increase the cost and duration of treatment.

Bacteremia↗
PubMed1992

[The cholera epidemic in Latin America].

An outbreak of cholera started in Peru in January 1991 and spread through most Latin American countries within a year. This was the first known epidemic of cholera in America for more than a century. In 1991, 321,334 persons were reported to have cholera in Peru, 119,063 were hospitalized, and 2,906 died. Other countries like Ecuador, Colombia, Guatemala, Brazil, Mexico, Bolivia, Chile, El Salvador, Venezuela and Honduras were also affected, but these countries combined accounted for only 20% of the cases registered in Peru. In April 1992, all Latin American countries except Uruguay, Paraguay and French Guyana have reported cholera. The mortality rate for the epidemic in Latin America was only 1%, mainly owing to good oral rehydration treatment provided by Local health services and the Pan American Health Organization. The causative organism was Vibrio cholerae, serogroup O1, serotype Inaba (and Ogawa) of the El Tor biotype. Genetic characterization shows this strain to be unique, and the designation is reserved for the Latin American strain, distinguishing it from the other El Tor isolates from the 7th pandemic.

Cholera↗
PubMed1992

Characterization of Escherichia coli strains isolated from pigs with edema disease.

Escherichia coli strains belonging to serogroups O 138 and O 139 isolated from pigs with edema disease, were characterized with respect to the presence of genes encoding Shiga-like toxin I, Shiga-like toxin II and Shiga-like toxin IIv (SLT I, SLT II and SLT IIv). Genes coding for the heat-stable and heat-labile enterotoxins (ST I and LT I) were also detected. Plasmid profiling, restriction enzyme digestion of total DNA, and ribotyping were performed for further characterization of the strains. The oligonucleotide probes applied in this study appeared to be useful tools for detecting genes coding cytotoxins and enterotoxins. DNA from 12 of 16 strains hybridized with two SLT II probes, and DNA from two SLT IIv encoding strains also hybridized with the ST I probe. DNA from one SLT IIv negative strain hybridized with the LT I probe. The results from plasmid profiling, restriction enzyme digestion, and ribotyping were compared with serogrouping in attempts to distinguish between the different E. coli edema disease isolates. Fourteen different plasmid profiles were identified, and as restriction patterns barely did, and ribotyping patterns did not, reveal any information useful for differentiation of the strains beyond serogroup level, plasmid profiling seemed to be the most suitable method for discrimination between the edema disease strains investigated here.

Animals↗
PubMed1992

Tetracycline resistance genes in Kenyan hospital isolates of Salmonella typhimurium.

All 97 strains of Salmonella typhimurium isolated from patients at a hospital in Nairobi, Kenya, during 1988-90 were resistant to tetracycline. The minimum inhibitory concentration (MIC) showed a large distribution range from 1 microgram/ml to 128 micrograms/ml. The strains were heterogeneous with respect to plasmid content, but initially all strains possessed, in addition to other plasmids, a large 60-, 63- or 65-MDa plasmid. The tetracycline resistance genes were characterized using oligonucleotide probes, and 20% of the resistant strains possessed tetracycline type A (tetr A), 6% tetr B, and 4% tetrC genes. Three strains possessed both type A and B tetracycline resistance determinants, which were shown to be located on the large 65-MDa plasmid. There was no correlation between strains isolated from stools, blood, cerebrospinal or epidural fluids, pus, or urine, with respect to the tetracycline genotypes, MIC values or plasmid content.

Base Sequence↗
PubMed1992

Use of polymerase chain reaction for detection of toxigenic Vibrio cholerae O1 strains from the Latin American cholera epidemic.

In January 1991, an outbreak of cholera started in Peru and spread throughout most of Latin America within 8 months. As of March 1992, over 450,000 cases and approximately 4,000 deaths have been reported to the Pan American Health Organization. The causative organism is toxigenic Vibrio cholerae O1 of the El Tor biotype and is distinct from the U.S. Gulf Coast strains. A polymerase chain reaction (PCR) that amplifies a 564-bp fragment of the cholera toxin A subunit gene (ctxA) was used to identify toxigenic V. cholerae O1 strains. A total of 150 V. cholerae O1 isolates were tested. They were of unknown toxin status, were associated with recent outbreaks, and were isolated from patients, food, and water. One hundred forty isolates were found to be toxigenic both by PCR and the routine diagnostic enzyme-linked immunosorbent assay. Thirty-eight known toxigenic strains isolated worldwide from 1921 to 1991 were also positive in the PCR. A collection of 18 nontoxigenic V. cholerae O1 strains, 35 Escherichia coli heat-labile-enterotoxin-I-producing strains, 26 Campylobacter strains, and 8 strains of Aeromonas hydrophila, previously reported to produce cholera toxin-like toxin, were all negative in the ctxA PCR. We conclude that this PCR is a diagnostic method that specifically detects toxin genes in V. cholerae O1 strains in a reference laboratory. It is more rapid and less cumbersome than other diagnostic methods for detection of toxicity in these strains.

Bacteriological Techniques↗