Bacteriocin Typing Research Articles

Taxonomy of Yersinia pestis.

This chapter summarized the taxonomy and typing works of Yersinia pestis since it's firstly identified in Hong Kong in 1894. Phenotyping methods that based on phenotypic characteristics, including biotyping, serotyping, antibiogram analysis, bacteriocin typing, phage typing, and plasmid typing, were firstly applied in classification of Y. pestis in subspecies level. And then, with the advancement of molecular biological technology, the methods based on outer membrane protein profiles, fatty acid composition, and bacterial mass fingerprinting were also used to identify the populations within Y. pestis. However, Y. pestis is a highly homogenous species; therefore, the above typing methods could only provide low resolution, e.g., only one serotype and one phage type were observed for the whole species. Since the 1990s, molecular typing based on DNA variations, including single-nucleotide polymorphism, gene gain/loss, variable-number tandem repeats, clustered regularly interspaced short palindromic repeat, etc., was introduced and improved the resolution and robust of typing result. Especially in recent years, genotyping-based whole-genome-wide variations were successfully employed in Y. pestis, which built the "gold standard" of typing scheme of the species and could distinguish the samples under the strain level. The taxonomy and typing works leaved us enormous polymorphism data; therefore, a comprehensive fingerprint database of Y. pestis was needed to collect and standardize these data, for facilitating future works on evolution, plague surveillance and control, anti-bioterrorism, and microbial forensic researches.

Screening bactericidal effect of Pectobacterium carotovorum subsp. carotovorum strains against causal agent of potato soft rot.

This study focuses on the potential of Pectobacterium carotovorum subsp. carotovorum (Pcc) strains producing bacteriocin as a tool to control potato soft rot disease. Thirty out of 48 purified bacterial strains were characterized as Pcc using specific PCR and phenotypic tests. The pathogenicity and pectate degrading assays were recorded positive for 13 strains. Bacteriocin typing clustered producers into three groups according to their antimicrobial spectra. Majority of the producers except strains of group II showed antibacterial activity toward relative genus and the role of UV or mitomycin C was inductive. In addition, none of the distant genus was sensitive to Pcc bacteriocins except Rhizobium vitis. Molecular detection of four bacteriocins including carotovoricin, carosin S1, S2 and carosin D was performed. Overall, 54.5% of group I, 47.3 and 70% of groups II and III strains carried carotovoricin and four strains harbored gene corresponding to carosin S1. According to our data divers antimicrobial patterns obtained by Pcc strains and existence of new bateriocines could be possible. Moreover, our findings recommended that direct application of P29 or expression of corresponding genes of Pog22 or P21 in a nonpathogenic strain as a biocontrol agent may improve soft rot disease control.

English

INTRODUCTION: Pseudomonas aeruginosa (P.aeruginosa) is a ubiquitous Organism that is often found in environment. It is a Gram-negative Bacillus, motile, straight (or) slightly curved measuring 0.5 – 8 Micrometer by 1.5 to 3.0 micrometer, non-spore forming, and Noncapsulated & obligate aerobes. Pseudomonas putida can appear Elongated. Organism from old culture may appear pleomorphic Gram Negative. Mucoid strains are seen as long filaments surrounded by Darker pink staining material (Alginate). Gram negative bacilli that are Showing intra cellular presence in polymorphonuclear leucocytes are Clinically important. Flagellar staining reveals single polar flagella. It Grows at wide temperature range between 6-42oc . Most pseudomonas species can reside in a wide variety of Environmental niches. Since their preferred temperature requirements Are between 4 and 36oc, Pseudomonas are found in processed foods, in Hands of health professional and they colonise animals and human. They Can spread among individuals. Ear infection and an irritating folliculitis may be acquired from Swimming pools, hot tubs. Community associated P. Aeruginosa Infections may occur as consequent to exposure to the organism in hot Tubs, whirlpools, and swimming pools. Keratitis due to P.aerugionsa May occur in individuals who store contact lenses in tap water and or Contaminated contact lens solutions. Intra venous drug users may Develop endocarditis and osteomyelitis as a result of injection of Contaminated injectables. P. Aeruginosa is also of nosocomial Significance The organism has been isolated from a variety of solutions Including soaps, ointments, irrigation and dialysis fluids, eye drops and Disinfectants as well as from fomites such as shower heads, respiratory Therapy unit, sinks, baths etc. Populations at risk for significant Morbidity and mortality include intubated patients in intensive care, Patients on chronic ambulatory peritoneal dialysis, and burns patients. Pseudomonas Species are associated with a number of Infections. They produce infections of wound and burns giving rise to Blue green pus. P. Aeruginosa can cause meningitis usually following Trauma (or) surgery. Corneal infections result from contaminated contact Lenses (or) injury, contaminated eye drops, and ointments. Pseudomonas Causes endocarditis through valve replacement. People who spend long periods for swimming are at risk for External ear infections (Swimmer’s ear). Malignant external otitis is a Virulent form of disease seen primarily in diabetes and elderly patients. P. Aeruginosa is the leading cause of chronic otitis media. Folliculltis is a common infection caused by Pseudomonas Species resulting from immersion in contaminated water (e.g. Hot tubs, Whirlpools, and swimming pools). Thermal burns of skin inhibit an Essential component of body defence against infection; the physical Barrier of intact skin .The resulting damaged tissue is a rich culture Medium and is a great risk for colonization and infection by Pseudomonas. Such infections have been the leading cause of morbidity And mortality. Infection of burn wound with Gram negative bacteria Typically occur one week after injury .The moist surface of the burnt Skin and lack of neutrophilic response to tissue invasion predispose Patients to such infections. Infections of the urinary tract are seen Primarily in patients with long-term indwelling catheter. The lungs of Children with cystic fibrosis are particularly susceptible to P. Aeruginosa. Use of ventilator which may introduce the organism to lower airways Produces lung abscess and pneumonia .The mortality rate is 70%. Epidemiological investigations of P. Aeruginosa infection have different Typing methods as follows 1 Biotyping 2. Antiobiotic Sensitivity Typing 3. Serotyping 4. Bacteriophage typing 5. Bacteriocin Typing 6. Modified Diene’s mutual Inhibition Test 7. Random Amplified Polymorphic DNA Study The Bacteriophage and Bacteriocin typing are outdated because Cross-reactions are more common. The Diene’s mutual inhibition test has been used as an Epidemiological tool to characterize Pseudomonas aeruginosa like Proteus mirabilis. The test was simple to perform and cheap and may Have utility in initial screening of P.aeruginosa isolates in suspected Common source epidemic. Genomic finger printing methods are now regarded as the most Accurate methods for the typing of microorganisms for epidemiological Purposes. These methods include pulse field gel electrophoresis, Ribotyping, and Polymerase chain reaction based finger printing (RAPD – PCR). In the present study, P.aeruginosa was isolated from heterogenous Clinical samples from patients attending Govt Rajaji Hospital, Madurai And were subjected to various tests to identify strain similarity. Further They were screened for similar homology using phenotypic test, Diene’s Mutual Inhibition test, and genotypic test RAPD – PCR.

Bacteriocin Typing of Staphylococcus Aureus Isolated from Different Sources in Ibb City , Yemen

A total of 207 specimens were collected from different sources including patients, health care staff and hospital environment in Ibb city, Yemen. The study used the bacteriocin produced from active producer strains in typing of Staphylococcus aureus. Depending on the morphological, cultural and biochemical characteristics, 54 (26.09%) isolates of Staphylococcus aureus were identified. An antibiotic sensitivity test was done for the bacterial isolates, and the results showed that there were multiple resistant antibiotics. The Staphylococcin production of these isolates has been detected by using wells assay. Fifty one isolates were Staphylococcin producer. Four isolates (staph19, staph25, staph28 and staph43) were chosen as good Staphylococcin producers, and used locally as indicators in bacteriocin typing. Depending on S. aureus typing, the isolates fell into (9) groups. The most numerous group was characterized by susceptibility to all four staphylococcin and comprised 61.11% isolates of S. aureus, while the lowest numerous were found in three groups with a ratio of 1.85%; the remaining groups had little percentages ranging from 3.70% to 11.11%. We observed that about (94.44%) of the isolates were bacteriocin producers, and among them, four isolates had a strong bacteriocin production. Based on typing, most isolates had one pattern.

Purification and characterization of streptin, a type A1 lantibiotic produced by Streptococcus pyogenes.

Approximately 10% of Streptococcus pyogenes strains inhibit the growth of all nine indicators in a standardized streptococcal bacteriocin typing scheme. The present study has shown that this inhibitory profile, referred to as bacteriocin producer (P)-type 777 activity, is due to the type A1 lantibiotic streptin. Two major forms of streptin were purified to homogeneity from 95% acidified (pH 2) methanol extracts of S. pyogenes M25 cells by using a series of reversed-phase chromatographic separations. The fully processed form of streptin (streptin 1) is a 23-amino-acid peptide with a mass of 2,424 Da. The 2,821-M(r) form of the peptide (streptin 2) has three additional amino acids (TPY) at the N terminus. Strain M25 extracts also contained small quantities of the streptin 1 and streptin 2 peptides in various stages of dehydration. Streptin 1 and streptin 2 were each capable of specifically inducing streptin production when added to strain M25 cultures. The streptin gene cluster resembled that of other type A1 lantibiotics but appeared to lack a streptin-specific proteinase gene. Although the streptin structural gene (srtA) was widespread within S. pyogenes, being detected in 40 of 58 strains, each representing a different M serotype, only 10 of these srtA-positive strains produced active streptin. The failure of some strains to express streptin was attributed to an approximately 4.5-kb deletion in their streptin loci, encompassing genes putatively encoding proteins involved in streptin processing (srtB and srtC) and transport (srtT). In other strains, srtA transcription appeared to be defective. No direct association could be detected between the production of streptin and the production of the lantibiotic-like hemolysin streptolysin S in strain M25.

Transferable plasmid mediating resistance to multiple antimicrobial agents in Klebsiella pneumoniae isolates in Greece

Objective To investigate the underlying resistance mechanisms in 10 Klebsiella pneumoniae isolates.Methods Ten K. pneumoniae strains according to distinct bacteriocin typing and REP-PCR, were examined for their plasmid content, their ability to transfer their resistance to aminoglycosides and third-generation cephalosporins, and their production of aminoglycoside-modifying enzymes and β-lactamases.Results Transfer of resistance to the above-mentioned antibiotics as well as to co-trimoxazole and tetracycline in Escherichia coli strain RC 85 at a frequency of 5–106 was achieved for all strains by conjugation. Similar strains harbor a self-transferable multiresistant plasmid (80 kb) with similar EcoRI and HindIII restriction patterns. This plasmid encodes an extended-spectrum β-lactamase which confers high-level resistance to third-generation cephalosporins and aztreonam. It produces SHV-5 β-lactamase, as demonstrated by isoelectric focusing and DNA sequencing. Aminoglycoside resistance was co-transferred, and AAC(6′)-I, mediating resistance to gentamicin, tobramycin, netilmicin and amikacin, and AAC(3)-I, mediating resistance to gentamicin and sisomycin, were encoded in all isolates and their transconjugants, while APH(3′)-I, mediating resistance to kanamycin and neomycin, was encoded in seven strains.Conclusions It appears that a multiresistant transferable plasmid encoding the SHV-5 β-lactamase, causing unusually high resistance to ceftazidime and aztreonam, and the combination AAC(6′)-I + AAC(3)-I of acetylating enzymes causing, also resistance to all clinically available aminoglycosides, is established in K. pneumoniae in Greece.

Production of bacteriocin by Bacteriodes fragilis and partial characterization.

The ability of Bacteroides fragilis strains, isolated from various sources, to produce bacteriocin was evaluated. All strains isolated from intestinal infections were producers in high levels and less susceptible to the others. Strains from other origins were found to produce bacteriocin at a medium level and they were variably susceptible. Some properties of one bacteriocin produced by the Bact. fragilis 079298-3 strain were analysed, providing evidence of its protein nature, with stability over a wide range of pH and retained inhibitory activity after heating. This variability seems to suggest that bacteriocin typing is a good method for this species.

Surveillance of Pseudomonas aeruginosa in Intensive Care Units: Clusters of Nosocomial Cross-Infection and Encounter of a Multiple-Antibiotic Resistant Strain

Serogrouping (determination of O antigens) and bacteriocin typing (based on susceptibility to one or more of 18 bacteriocins) were employed to survey 210 isolates of Pseudomonas aeruginosa from 201 patients in 8 intensive care units (ICU) during an observation period of 18 months. Eighty-eight isolates (41.9%) were nonserogroupable (NT); most common were serogroups O1, O9, O11, and O3. All except 5 isolates (97.6%) were bacteriocin-typable. However, phenotypic variation of bacteriocin susceptibility, in particular the receptor for bacteriocin No. 13, rendered this typing method presumptive as well. Bacteriocin susceptibility profiles were not predictive of serogroup and vice versa. Workup of 19 isolates from 9 patients disclosed phenotypic variation of antibiotic susceptibility in 3 patients, superinfection by a different strain in 4 patients, and persistence (3 months) of the same strain in 2 patients, respectively. Serotyping and bacteriocin susceptibility test data revealed 15 clusters of putative cross-infection of 2 patients each, 8 clusters involving 3 patients each, one outbreak (serogroup NT, bacteriocin profile 777736) involving 4 patients in the pediatric ICU, one outbreak due to a multiple-antibiotic resistant (MAR) strain in the surgical ICU (4 patients, serogroup O12, bacteriocin profile 30400), and two putative outbreaks in the pneumonology ICU involving 6 patients (serogroup NT, bacteriocin profile 777726) and 9 patients (serogroup NT, bacteriocin profile 777736). Pulsed-field gel electrophoresis (PFGE) macrorestriction analysis (SpeI, XbaI) confirmed the pediatric and surgical ICU strains as singular strains. However, the two putative outbreaks in the pneumonology ICU were due to one particular strain which had infected 13 of the 15 patients as determined with the PFGE genotypic method. Isolates comprising the MAR strain of P. aeruginosa were susceptible only to amikacin, fosfomycin, and polymyxin B; the isolates varied in susceptibility to aztreonam and ceftazidime. This MAR strain was susceptible to the bactericidal activity of 65 vol% of fresh defibrinated human blood from donors B, L, and T. Either amikacin (16 µg/ml) or fosfomycin (8 µg/ml) plus blood and amikacin (8 µg/ml) combined with fosfomycin (8 µg/ml) with and without blood consistently killed isolates of the MAR strain, which thus was amenable to antibiotic therapy.

Mutacin production by Streptococcus mutans may promote transmission of bacteria from mother to child.

The production of bacteriocin-like inhibitory substances, mutacins, by mutans streptococci varies among isolates. To find if the degree of mutacin activity of an isolate was related to its transmission between mother and her child, 19 mothers and their 18-month- to 3-year-old children were sampled for their oral mutans streptococci. In addition, the stability of mutacin activity was studied with isolates from the mothers and with isolates from five unrelated 5-year-old children in 5- to 7-year follow-up studies. A total of 145 oral mutans streptococcal isolates were serotyped by immunodiffusion, ribotyped, and mutacin typed by the stab culture technique. Mutacin was produced by 88% of the strains against more than 1 of the 14 indicator strains, representing mutans streptococci, Streptococcus sanguis, Streptococcus salivarius, Streptococcus oralis, Streptococcus gordonii, and Streptococcus pyogenes. Streptococcus mutans isolates showed more inhibitory activity than did Streptococcus sobrinus isolates. Identical ribotypes had similar mutacin activity profiles within a subject, initially and in the follow-up studies, in all but two cases. The mothers harbored a total of 37 different mutans streptococcal ribotypes. Six children were negative for mutans streptococci. Transmission was probable in 9 of 20 mother-child pairs on the basis of the presence of identical strains, as determined by ribotyping and bacteriocin (mutacin) typing. S. mutans strains shared between a mother and her child showed a broader spectrum of inhibitory activity than did nontransmitted strains. In conclusion, the mutacin activity of clinical isolates is reasonably stable, and this virulence factor seems to be of clinical importance in early colonization by S. mutans.

Strain differentiation of Clostridium perfringens by bacteriocin typing, plasmid profiling and ribotyping.

Bacteriocin typing, plasmid profiling and ribotyping were used to type 34 food and patient Clostridium perfringens isolates from 10 food poisoning cases, respectively, outbreaks. In nine cases/outbreaks bacteriocin patterns showed identical main groups. Subgroups differed within all cases/outbreaks. Plasmid profiles were identical for all isolates within each of three outbreaks. In eight food poisoning cases and outbreaks, all the ribotypes of each food and stool isolate were found to be identical. All three typing methods give valuable results for the characterization of C. perfringens beyond the species level. Bacteriocin typing represents a suitable addition to plasmid typing, particularly since the results do not show any correlation between losses of plasmids and changes in bacteriocin sensitivity patterns. Ribotyping was found to be a suitable tool to determine the genetic relationship of C. perfringens isolates in the context of food-borne poisoning.

Multiresistant Pseudomonas aeruginosa serogroup O:11 outbreak in an intensive care unit.

OBJECTIVE: To determine whether 15 multiresistant Pseudomonas aeruginosa isolates from an intensive care unit (ICU) outbreak were related, were endemic, and belonged to the O:12 European clone. METHODS: Forty-six P. aeruginosa isolates from a large hospital were investigated with respect to their antibiotic resistance profiles, serogroups, bacteriocin types and DNA fingerprints obtained by pulsed-field gel electrophoresis (PFGE) of genomic DNA digested with Xbal. RESULTS: Fourteen of the ICU outbreak isolates were indeed identical with respect to their serogroup, O:11, pyocin type, 10/a, and PFGE type, A. Clone A was endemic and dominant throughout the hospital, even though, within the ICU, it underwent phenotypic alterations, such as loss of cell wall lipopolysaccharide side-chains, or acquisition of ceftazidime and imipenem resistance. Bacteriocin typing was more discriminatory than serotyping, but PFGE could differentiate further among phenotypically identical strains. It also allowed the tracking of an O:6 strain, as it was becoming gradually more resistant and undergoing a bacteriocin-type conversion while remaining genotypically unaltered. CONCLUSIONS: Using three typing methods, a nosocomial multiresistant strain distinct from the previously described dominant European O:12 clone was characterized, and the ability of PFGE to identify clonal isolates even when these appear phenotypically distinct was demonstrated.

Serratia marcescens.

Over the last 30 years, Serratia marcescens has become an important cause of nosocomial infection. There have been many reports concerning the identification, antibiotic susceptibility, pathogenicity, epidemiological investigations and typing of this organism. Accurate identification is important in defining outbreaks. The API 20E system has been used widely, but is not individually satisfactory. The growth of S. marcescens in the environment has been investigated in relation to water, disinfectants and plastics such as blood bags. Certain extracellular products are unique to S. marcescens. Pigment (prodigiosin) biosynthesis by S. marcescens has been investigated fully since the emergence of the organism as a cause of infection. Many other aspects of the pathogenicity and virulence of S. marcescens have been studied, including adherence and hydrophobicity, lipopolysaccharide (LPS) and extracellular products. Two modes of adhesion to host epithelial surfaces have been suggested. These are mannose-resistant (MR) pili and mannose-sensitive (MS) pili. LPS, which is responsible for the biological activity of endotoxin, has been investigated fully and 24 somatic antigens have been described. The production of different enzymes by S. marcescens as virulence factors has also been reported, including chitinase, lipase, chloroperoxidase and an extracellular protein, HasA. Antibiotics used to treat serratia infection include beta-lactam agents, aminoglycosides and fluoroquinolones and a variety of different resistance mechanisms have been demonstrated. Typing methods used to study the epidemiology of S. marcescens include biotyping, bacteriocin typing, phage typing, plasmid analysis, polymerase chain reaction amplification of enterobacterial repetitive intergenic consensus sequences (ERIC-PCR) and ribotyping. Serological typing has also been used and this method seems to be a suitable first-line typing method for S. marcescens, although some strains remain untypable. RAPD-PCR has also been applied to a small number of isolates and seems to be a promising method, especially for rapid monitoring of an outbreak and tracing the source of initial infection.

Strain Differentiation and Taxonomic Characterisation of a Thermophilic Group of Phenol-degrading Bacilli

Thirteen new thermophilic phenol degrading strains were isolated from a number of geographically distinct thermal sources. The strains demonstrated similar growth characteristics with pH optima between 6.0–7.2, temperature optima between 60 °C–70 °C and similar substrate utilisation spectra. The comparison of chromosomal restriction endonuclease, ribotyping and total cellular protein patterns in addition to bacteriocin typing was used for strain differentiation and resulted in the formation of five different groups to which the isolates could be assigned. Eight of the isolates were assigned to group 2 based on identical patterns generated for each method applied. Additional methods such as the comparison of antibiotic MIC values, phenol tolerance, co-metabolic transformation of chlorophenols, fatty acid methyl ester (FAME) patterns and RFLP patterns revealed minor differences between the isolates of group 2. Isolate A2, of this group and A7, which displayed differences using every technique applied, were selected for taxonomic identification and were identified as B. thermoleovorans and B. thermoglucosidasius respectively. This is the first report of phenol degrading isolates belonging to these species.

An Introduction to the Hows and Whys of Molecular Typing

Until recently, the relatedness of bacterial isolates has been determined solely by testing for one or several phenotypic markers, using methods such as serotyping, phage typing, biotyping, antibiotic susceptibility testing, and bacteriocin typing. However, there are problems in the use of many of these phenotype-based methods. For example, phage and bacteriocin typing systems are not available for all bacterial species and serotyping can be labor-intensive and costly. In addition, phenotypic markers may not be stably expressed under certain environmental or culture conditions. In contrast, some of the newer molecular typing methods involving the analysis of DNA offer many advantages over traditional techniques. One of the more important advantages is that since DNA can always be extracted from bacteria, all bacteria should be typeable. Another is that the discriminatory power of DNA-based methods is greater than that of phenotypic procedures. This review focuses on the basics of molecular typing along with the advantages and disadvantages of several of the newer genotypic typing techniques. This includes methods such as plasmid typing, pulsed-field gel electrophoresis, ribotyping and its variations, and polymerase chain reaction-based methods such as random amplified polymorphic DNA analysis. Molecular typing of microorganisms has made great strides in the last decade, and many food microbiology laboratories have become more knowledgeable and better equipped to carry out these new molecular techniques. Molecular typing procedures can be broadly defined as methods used to differentiate bacteria, based on the composition of biological molecules such as proteins, fatty acids, carbohydrates, etc., or nucleic acids. The latter can also be more specifically defined as genotyping, and is the subject of this review.

Typing of nosocomial Strains of Serratia marcescens: Comparison of restriction enzyme cleaved genomic DNA fragment (PFGE) analysis with bacteriocin typing, biochemical profiles and serotyping

Eighty-eight selected clinical isolates of Serratia marcescens, representing 27 putative outbreaks of nosocomial cross-infection encountered during 1980-1995, were tested comparatively by bacteriocin typing, carbon source assimilation tests, serotyping (O and H antigens), and restriction pattern (RFLP) analysis of restriction cleaved (SpeI, XbaI) genomic DNA fragments after pulsed-field gel electrophoresis (PFGE). Serotyping served as the "gold standard" of the phenotypic methods. One pseudo-outbreak (bacteriocin typing incriminated type 26) was uncovered through serotyping as well as the biochemical profile and confirmed by PFGE analysis of genomic DNA. Bacteriocin typing and determination of biochemical profiles disclosed several instances of phenotypic variation; serotyping revealed two episodes of shifts from motility (H12) to nonmotility. Resolution of restricted genomic DNA fragments with the PFGE procedure permitted detection of 27 PFGE patterns (A-M, N-1-N-3, O-1, O-2, P-1-P-3, Q-1-Q-3, R-1-R-3, S-1, and T). Based on the analysis of PFGE patterns against the background of epidemiological data, the number of nosocomically significant strains of S. marcescens could be reduced to 16 (PFGE patterns A-M, N-2, O-1, P-2, and T). It was concluded that PFGE analysis of restricted genomic DNA of S. marcescens was superior to the three phenotypic methods.

Phenotypic variation of clinical Serratia marcescens isolates repeatedly recovered from individual patients

A total of 129 selected isolates of Serratia marcescens which had been recovered from 50 patients during the 1980-1995 period and which revealed phenotypic variation in terms of bacteriocin (phage tail) susceptibility, carbon source assimilation, or serotype, were reexamined with these three phenotypic methods. Seven isolates (5.4%) were bacteriocin nontypable; all 129 isolates utilized carbon sources and could be serotyped. Fourty-eight isolates from 20 patients yielded unambiguous results with these 3 phenotypic methods and were excluded from further analysis. Among the remaining 81 isolates from 30 patients, isolates from 2 patients revealed phenotypic variation in bacteriocin susceptibility only, whereas isolates from 6 patients showed variant bacteriocin types and variant biochemical profiles, but were of identical serotype. Isolates from 20 patients revealed variant biochemical profiles only. Three patients had become superinfected with strains of S. marcescens of different phenotype and genotype. In 4 patients, previously motile (H12) isolates had become nonmotile (H-). PFGE analysis of XbaI and SpeI-restricted genomic DNA of the 81 isolates of the 30 patients demonstrated the isolates of 22 patients to be genotypically identical. The isolates from 3 patients were closely related by genotype, and those from an additional patient proved to be possibly related. PFGE analysis demonstrated one patient to have become infected by two genotypically different strains of S. marcescens of identical serotype, which, however, differed in bacteriocin type and biochemical profile. It was concluded that PFGE analysis of restricted genomic S. marcescens DNA was superior to the three phenotypic methods examined comparatively. Serotyping was more reliable than bacteriocin typing, and the latter technique yielded fewer phenotypic variants than determination of biochemical profiles among consecutively recovered isolates from patients with long-lasting S. marcescens infection.

Bacteriocin Typing of Burkholderia (Pseudomonas) solanacearum Race 1 of the French West Indies and Correlation with Genomic Variation of the Pathogen

Burkholderia solanacearum race 1 isolates indigenous to the French West Indies were characterized by bacteriocin typing and two genomic fingerprinting methods: pulsed-field gel electrophoresis of genomic DNA digested by rare-cutting restriction endonucleases (RC-PFGE) and PCR with primers corresponding to repetitive extragenic palindromic (REP), enterobacterial repetitive intergenic consensus (ERIC), and BOX elements (collectively known as rep-PCR). The survey comprised 24 reference strains and 65 isolates obtained from a field trial in Guadeloupe in 1993. Comparison of the data identified RC-PFGE as the most discriminatory method, delineating 17 pulsed-field gel profile types. rep-PCR and bacteriocin typing identified nine rep-PCR profile types and nine bacteriocin groups. Independent determination of similarity coefficients and clustering of RC-PFGE and rep-PCR data identified six groups common to both sets of data that correlated to biovar and bacteriocin groups. Further study of bacteriocin production in planta gave results consistent with in vitro bacteriocin typing. It was observed that spontaneous bacteriocin-resistant mutants exhibited a cross-resistance to other bacteriocins as identified by the typing scheme and that such mutants possessed a selective advantage for growth over isogenic nonmutants in the presence of a bacteriocin. The results are significant in the search for biological control of disease by nonpathogenic mutants of the wild-type organism.

Taxonomy and Phylogeny of Industrial Solvent-Producing Clostridia

We performed a systematic study of 55 solvent-producing clostridial strains, the majority of which are currently classified as Clostridium acetobutylicum strains, by using a combination of biotyping and DNA fingerprint analysis. The biotyping procedures used included rifampin susceptibility testing, bacteriocin typing, and bacteriophage typing. The 55 strains examined exhibited a good correlation between their biotypes and DNA fingerprints, which allowed us to divide them into nine groups. The DNA fingerprints of the nine groups differed markedly, but within each group the DNA fingerprints exhibited a high level of similarity. To determine the phylogenetic relationships of the nine groups, we performed a 16S rRNA gene sequence analysis. The results of a comparative analysis of the partial sequence corresponding to positions 830 to 1383 (Escherichia coli numbering) of the 16S rRNA gene indicated that the nine biotype groups could be assembled into four taxonomic groups. The complete 16S rRNA sequences of strains representing these groups were determined. Our phylogenetic analysis revealed that the amylolytic type strain C. acetobutylicum ATCC 824 (taxonomic group I) was only distantly related to the saccharolytic strains belonging to taxonomic groups II, III, and IV (levels of sequence similarity, 90 to 90.5%). The strains belonging to taxonomic groups II, III, and IV, represented by C. acetobutylicum NCP 262, "Clostridium saccharoperbutylacetonicum" N1-4, and C. acetobutylicum NCIMB 8052T (T = type strain), respectively, were closely related (levels of sequence similarity, 98.2 to 98.9%). C. acetobutylicum NCIMB 8052T exhibited a level of similarity of 100% with the type strain of Clostridium beijerinckii. Reclassification of the saccharolytic solvent-producing strains is necessary, and possible names for the four taxonomic groups are discussed.

An Outbreak of Pseudomonas aeruginosa Ventilator-Associated Respiratory Infections Due to Contaminated Food Coloring Dye: Further Evidence of the Significance of Gastric Colonization Preceding Nosocomial Pneumonia

An outbreak of ventilator-associated Pseudomonas aeruginosa respiratory infections in an intensive care unit of a teaching hospital was found to be associated with contaminated food coloring dye. Serotyping and bacteriocin typing indicated that the same strain of Pseudomonas was isolated from the food dye and from the respiratory cultures of the majority of patients. This observation lends credence to the importance of gastric colonization preceding pneumonia in ventilated patients.

An outbreak of Pseudomonas aeruginosa ventilator-associated respiratory infections due to contaminated food coloring dye--further evidence of the significance of gastric colonization preceding nosocomial pneumonia.

An outbreak of ventilator-associated Pseudomonas aeruginosa respiratory infections in an intensive care unit of a teaching hospital was found to be associated with contaminated food coloring dye. Serotyping and bacteriocin typing indicated that the same strain of Pseudomonas was isolated from the food dye and from the respiratory cultures of the majority of patients. This observation lends credence to the importance of gastric colonization preceding pneumonia in ventilated patients.