Analysis Of Whole-cell Protein Patterns Research Articles

Diversity and enzymatic potential of thermophilic bacteria associated with terrestrial hot springs in Algeria.

This study aims to determine the diversity of culturable thermophilic bacteria isolated from eight terrestrial hot springs in Northeastern of Algeria using the conventional methods, SDS-PAGE fingerprinting of whole-cell proteins and 16S rRNA gene sequencing. In addition, their hydrolytic enzyme activities were also investigated. A total of 293 strains were isolated from the hot springs' water and sediment using different culture media. Overall, five distinct bacterial groups were characterized by whole-cell protein pattern analysis. Based on the 16S rRNA gene sequencing of 100 selected strains, the isolates were assigned to the following three major phyla: Firmicutes (93%), Deinococcus-Thermus (5%), and Actinobacteria (2%), which included 27 distinct species belonging to 12 different phylotypes, Aeribacillus, Aneurinibacillus, Anoxybacillus, Bacillus, Brevibacillus, Geobacillus, Laceyella, Meiothermus, Saccharomonospora, Thermoactinomyces, Thermobifida, and Thermus. The screening for nine extracellular enzymes showed that 65.87% of the isolates presented at least five types of enzyme activities, and 6.48% of strains combined all tested enzymes (amylase, cellulase, pectinase, esculinase, protease, gelatinase, lipase, lecithinase, and nuclease). It was found that Bacillus, Anoxybacillus, Aeribacillus, and Aneurinibacillus were the genera showing the highest activities. Likewise, the study showed an abundant and diverse thermophilic community with novel taxa presenting a promising source of thermozymes with important biotechnological applications. This study showed that a combined identification method using SDS-PAGE profiles of whole-cell proteins and subsequent 16S rRNA gene sequence analysis could successfully differentiate thermophilic bacteria from Algerian hot springs.

Detection and Identification of Vibrio Species Using Whole-Cell Protein Pattern Analysis

Outbreaks of foodborne diseases associated with Vibrio species such as V. parahaemolyticus, V. vulnificus, and V. cholerae frequently occur in countries having a dietary habit of raw seafood consumption. For rapid identification of different Vibrio species involved in foodborne diseases, whole-cell protein pattern analysis for 13 type strains of 12 Vibrio species was performed using SDS-PAGE analysis. Pathogenic Vibrio species such as V. parahaemolyticus, V. vulnificus, V. cholerae, V. alginolyticus, V. fluvialis, and V. mimicus were included in the 12 Vibrio species used in this study. Each of the 12 Vibrio species showed clearly specific band patterns of its own. Two different strains of V. parahaemolyticus showed two different SDS-PAGE wholecell protein patterns, giving the possibility of categorizing isolated strains in the same V. parahaemolyticus species into two subgroups. The 36 Vibrio isolates collected from sushi restaurants in Busan were all identified as V. parahaemolyticus by comparing their protein patterns with those of Vibrio type strains. The identified isolates were categorized into two different subgroups of V. parahaemolyticus. The whole-cell protein pattern analysis by SDS-PAGE can be used as a specific, rapid, and simple identification method for Vibrio spp. involved in foodborne diseases at the subspecies level.

Burkholderia bannensis sp. nov., an acid-neutralizing bacterium isolated from torpedo grass (Panicum repens) growing in highly acidic swamps

Two strains of acid-neutralizing bacteria, E25(T) and E21, were isolated from torpedo grass (Panicum repens) growing in highly acidic swamps (pH 2-4) in actual acid sulfate soil areas of Thailand. Cells of the strains were gram-negative, aerobic, non-spore-forming rods, 0.6-0.8 µm wide and 1.6-2.1 µm long. The strains showed good growth at pH 4.0-8.0 and 17-37 °C. The organisms contained ubiquinone Q-8 as the predominant isoprenoid quinone and C(16 : 0), C(17 : 0) cyclo and C(18 : 1)ω7c as the major fatty acids. Their fatty acid profiles were similar to those reported for other Burkholderia species. The DNA G+C content of the strains was 65 mol%. On the basis of 16S rRNA gene sequence similarity, the strains were shown to belong to the genus Burkholderia. Although the calculated 16S rRNA gene sequence similarity of E25(T) to strain E21 and the type strains of Burkholderia unamae, B. tropica, B. sacchari, B. nodosa and B. mimosarum was 100, 98.7, 98.6, 97.6, 97.4 and 97.3 %, respectively, strains E25(T) and E21 formed a group that was distinct in the phylogenetic tree; the DNA-DNA relatedness of E25(T) to E21 and B. unamae CIP 107921(T), B. tropica LMG 22274(T), B. sacchari LMG 19450(T), B. nodosa LMG 23741(T) and B. mimosarum LMG 23256(T) was 90, 42, 42, 42, 45 and 35 %, respectively. The results of physiological and biochemical tests including whole-cell protein pattern analysis allowed phenotypic differentiation of these strains from previously described Burkholderia species. Therefore, strains E25(T) and E21 represent a novel species, for which the name Burkholderia bannensis sp. nov. is proposed. The type strain is E25(T) ( = NBRC 103871(T) = BCC 36998(T)).

Burkholderia acidipaludis sp. nov., aluminium-tolerant bacteria isolated from Chinese water chestnut (Eleocharis dulcis) growing in highly acidic swamps in South-East Asia

Two strains of aluminium-tolerant bacteria, SA33(T) and 7A078, were isolated from Chinese water chestnut (Eleocharis dulcis) growing in highly acidic swamps (pH 2-4) in actual acid sulfate soil areas of Vietnam (SA33(T)) and Thailand (7A078). The strains were Gram-negative, aerobic, non-spore-forming rods, 0.6-0.7 mum wide and 1.3-1.7 mum long. These strains showed good growth at pH 3.0-8.0 and 17-37 degrees C. The organisms contained ubiquinone Q-8 as the predominant isoprenoid quinone and C(16 : 0), C(18 : 1) ω 7c and C(17 : 0) cyclo as the major fatty acids. Their fatty acid profiles were similar to those reported for other Burkholderia species. The DNA G+C content of these strains was 64 mol%. On the basis of 16S rRNA gene sequence similarity, the strains were shown to belong to the genus Burkholderia. Although the 16S rRNA gene sequence similarity values calculated for strain SA33(T) to 7A078 and the type strains of Burkholderia kururiensis, B. sacchari and B. tuberum were 100, 97.3, 97.1 and 97.0 %, respectively, strains SA33(T) and 7A078 formed a group that was distinct in the phylogenetic trees; the DNA-DNA relatedness of strain SA33(T) to 7A078 and these three type strains were respectively 90, 47, 46 and 45 %. The results of physiological and biochemical tests, including whole-cell protein pattern analysis, allowed phenotypic differentiation of these strains from described Burkholderia species. Therefore, strains SA33(T) and 7A078 represent a novel species, for which the name Burkholderia acidipaludis sp. nov. is proposed. The type strain is SA33(T) (=NBRC 101816(T) =VTCC-D6-6(T)). Strain 7A078 (=NBRC 103872 =BCC 36999) is a reference strain.

Burkholderia heleia sp. nov., a nitrogen-fixing bacterium isolated from an aquatic plant, Eleocharis dulcis, that grows in highly acidic swamps in actual acid sulfate soil areas of Vietnam

Nitrogen-fixing bacteria, strains SA41(T), SA42 and SA53, were isolated from an aquatic plant, Eleocharis dulcis, that grows in highly acidic swamps (pH 2-4) in actual acid sulfate soil areas of Vietnam. The isolates were Gram-negative, aerobic, non-spore-forming, rod-shaped bacteria, having a cell width of 0.6-0.7 microm and a length of 1.5-1.7 microm. They showed good growth between pH 3.0 and 7.0, and between 17 and 37 degrees C. The organisms contained ubiquinone Q-8 as the predominant isoprenoid quinone, and C(16 : 0), C(17 : 0) cyclo, C(18 : 1) omega7c and summed feature 3 (C(16 : 1) omega7c and/or iso-C(15 : 0) 2-OH) as major fatty acids. Their fatty acid profiles are similar to those reported for other Burkholderia species. The DNA G+C content of these strains was 64 mol%. On the basis of 16S rRNA gene sequence similarity, these strains were shown to belong to the genus Burkholderia. Although their calculated 16S rRNA gene sequence similarity values to Burkholderia silvatlantica, Burkholderia mimosarum, Burkholderia ferrariae and Burkholderia tropica were 98.5, 98.2, 98.0 and 97.0 %, respectively, the isolates formed a distinct group in phylogenetic trees, and the DNA-DNA relatedness values of strain SA41(T) to these species were 39, 41, 39 and 33 %, respectively. The results of physiological and biochemical tests, including whole-cell protein pattern analysis, allowed phenotypic differentiation of these strains from the published Burkholderia species. Therefore, strains SA41(T), SA42 and SA53 represent a novel species for which the name Burkholderia heleia sp. nov. is proposed. The type strain is SA41(T) (=NBRC 101817(T)=VTCC-D6-7(T)).

Discrimination and numerical analysis of human pathogenic Candida albicans strains based on SDSPAGE protein profiles

In the present study, 21 Candida albicans strains were investigated using the commercial kit API 20C AUX system and the numerical analysis of whole-cell protein profiles. The results of the commercial kit confirmed that the all the strains belonged to C. albicans species. However, the research indicated that SDS-PAGE of polypeptides of whole-cell extracts can provide more valuable taxonomic information than conventional yeast test kits at the subspecies level. Despite the fact that C. albicans subtypes isolated from different anatomical sites had similar protein profiles, there were some distinctive protein bands. Numerical analysis of whole-cell protein profiles of all strains revealed 2 major clusters at similarity degrees of between 46.26 and 100%. Moreover, the results of numerical analysis confirmed that each cluster had characteristic and distinctive protein profiles. The research showed that, the morphological examination of yeast isolates remains essential to obtaining a correct identification, both the commercial yeast kit system and the numerical analysis of whole-cell protein patterns can be useful for the more reliable identification of C. albicans strains.

Angular Leaf Spot of Cucumber Caused by Pseudomonas syringae pv. lachrymans in Kurdistan.

During the spring of 2004 and 2005, angular leaf spot was observed on cucumber (Cucumis sativus L.) in some areas of Kurdistan Province in Iran for the first time. Disease incidence in affected fields was approximately 100%. Symptoms were initially small, round or irregular, water-soaked spots on leaves ranging from 3 to 5 mm in diameter. These spots were limited by the leaf veins that gave them an angular appearance. Under humid conditions, tiny, white exudates formed on the undersides of the leaves and severely infected leaves turned yellow. A fluorescent pseudomonad was consistently isolated from lesions on King's medium B and characterized. Twelve strains were selected from Marivan, Dehgolan and Kamyaran in western Iran. When compared with previously identified strains, (1,2) on the basis of phenotypic, biochemical, and physiological properties, isolates were identified as Pseudomonas syringae pv. lachrymans. This was confirmed with data from whole-cell protein pattern analysis, which indicated that the strains were highly similar to reference strain 4963T (International Collection of Microorganism from Plants [ICMP]). Isolates produced round, white colonies that were 1 to 2 mm in diameter. All strains were gram negative, aerobic, levan positive, oxidase negative, potato soft rot negative, arginine dihydrolase negative, and induced a hypersensitive response on tobacco leaves. The strains were positive for catalase, urease, and 4% NaCl tolerance and negative for nitrate reduction, methyl red production, acetoin and indole production, phosphatase, gas from glucose, reducing substances from sucrose, and ketolactose tests. All strains hydrolyzed Tween 80, esculin, casein, and gelatin, but failed to hydrolyze starch and lecithin. Results for growth at 41°C and production of hydrogen sulfide from cystein and peptone were negative. In Ayers' medium, all strains produced acid from d-galactose, citrate, sucrose, raffinose, fructose, d-xylose, glucose, inositol, mannitol, sorbitol, glycerin, mannose, ribose but not from trehalose, maltose, salicin, l-rhamnose, adonitol, cellobiose, ethanol, l-sorbose, inulin, dulcitol, starch, lactose or melibiose. All strains used l-asparagine, l-lysine, aspartate and l-arginine but did not use l-tartrate, propionate, ornithine, l-tyrosine, borate, benzoate, l-tryptophan, or acetate as carbon sources. Pathogenicity of four strains was confirmed by injecting bacterial suspensions (108 CFU/ml) into the undersides of 3- to 4-week-old cucumber leaves using a sterile syringe. Sterile water was injected into cucumber leaves as a negative control. Inoculated plants were maintained in the greenhouse at 25 to 28°C with 90 to 98% relative humidity until symptoms were assessed 4 to 7 days after inoculation. Angular leaf lesions developed on inoculated cucumber. Bacterial strains were reisolated from infected tissues and confirmed as P. syringae pv. lachrymans by biochemical characterization as previously described. This disease has been observed in other areas of Iran, but to our knowledge, this is the first report that characterizes the phenotypic and biochemical properties of the bacterium and disease in Kurdistan Province.

Burkholderia tropica sp. nov., a novel nitrogen-fixing, plant-associated bacterium

In an ecological survey of nitrogen-fixing bacteria isolated from the rhizosphere and as endophytes of sugarcane, maize and teosinte plants in Brazil, Mexico and South Africa, a new phylogenetically homogeneous group of N(2)-fixing bacteria was identified within the genus Burkholderia. This polyphasic taxonomic study included microscopic and colony morphology, API 20NE tests and growth on different culture media at different pH and temperatures, as well as carbon source assimilation tests and whole-cell protein pattern analysis. Analysis of 16S rRNA gene sequences showed 99.2-99.9 % similarity within the novel species and 97.2 % similarity to the closest related species, Burkholderia sacchari. The novel species was composed of four distinct amplified 16S rDNA restriction analysis groups. The DNA-DNA reassociation values within the novel species were greater than 70 % and less than 42 % for the closest related species, B. sacchari. Based on these results and on many phenotypic characteristics, a novel N(2)-fixing species is proposed for the genus Burkholderia, Burkholderia tropica sp. nov., with the type strain Ppe8(T) (=ATCC BAA-831(T)=LMG 22274(T)=DSM 15359(T)). B. tropica was isolated from plants grown in geographical regions with climates ranging from temperate subhumid to hot humid.

Enterococcus hermanniensis sp. nov., from modified-atmosphere-packaged broiler meat and canine tonsils.

Isolates 302, 334, 356, 377 and 379, detected in modified-atmosphere-packaged broiler meat, together with strains LMG 12317T and LMG 13617, detected in dog tonsils, were analysed in a polyphasic taxonomy study, including numerical analysis of ribopatterns and whole-cell protein patterns, 16S rRNA gene sequence analysis, DNA-DNA hybridization and determination of some phenotypic properties. The results indicated that these isolates represent a novel species in the genus Enterococcus. The isolates showed classical phenotypic reactions for the genus Enterococcus with the exception of not possessing the Lancefield group D antigen. Isolates 334, LMG 12317T and LMG 13617 showed the highest 16S rRNA gene sequence similarity (98.3-99.0 %) to the Enterococcus pallens type strain. In the distance matrix tree based on 16S rRNA gene sequences, the three isolates were located in the Enterococcus avium group with E. pallens as their closest phylogenetic neighbour. Numerical analyses of whole-cell protein patterns and HindIII/EcoRI ribotypes placed all seven isolates together in a single cluster separated from the E. avium group reference strains. The DNA-DNA hybridization level between strains 334 and LMG 12317T was 93.5 %, confirming that they represent the same species. Low hybridization levels (12-30 %) were, by contrast, obtained with the E. pallens and Enterococcus raffinosus type strains. The name Enterococcus hermanniensis sp. nov. is proposed, with strain LMG 12317T (= CCUG 48100T) as the type strain.

Weissella kimchii sp. nov., a novel lactic acid bacterium from kimchi.

A gram-positive, catalase-negative, non-sporulating, facultatively anaerobic, short rod-shaped bacterium, with cells measuring 0.3-0.5 x 1-2 microm and designated strain CHJ3T, was isolated from partially fermented kimchi, a traditional Korean fermented vegetable food. The strain produced CO2 gas, D-lactate from glucose and dextran from sucrose and hydrolysed aesculin and arginine. It also fermented N-acetylglucosamine, amygdalin, arbutin, cellobiose, D-fructose, galactose, beta-gentiobiose, gluconate, D-glucose, maltose, D-mannose, salicin, sucrose and D-xylose. The G+C content of the DNA was 48.2 mol%. Phylogenetic analysis of 16S rRNA showed that strain CHJ3T is a member of the genus Weissella. The nearest phylogenetic relative of strain CHJ3T was Weissella confusa, with 16S rRNA similarity of 98.3%. However, strain CHJ3T could be differentiated from W. confusa on the basis of some phenotypic characteristics, analysis of whole-cell protein patterns and DNA-DNA hybridization data. These data suggest that strain CHJ3T be classified in the genus Weissella as a novel species, Weissella kimchii sp. nov. The type strain is CHJ3T (= KCCM 41287T = DSM 14295T = KCTC 3746T).

Isolation and characterization of Lactococcus piscium strains from vacuum-packaged refrigerated beef.

To characterize gram-positive, catalase-negative, psychrotrophic, lactic acid-homofermentative, non-motile cocci isolated from vacuum-packaged refrigerated beef using phenotypic and genotypic methods. A total of 89 strains was isolated at 2 and 6 weeks as one of the predominant microflora of five samples of vacuum-packaged beef stored at 2 degrees C. The strains were compared with reference strains of some gram-positive, catalase-negative cocci using SDS-PAGE whole-cell protein pattern analysis, biochemical characterization and 16S rDNA sequencing. The biochemical and physiological characteristics of the isolates resembled those of Lactococcus piscium GTC 552(T). Numerical analysis of the SDS-PAGE whole-cell protein patterns resulted in close clustering of the strains with L. piscium GTC 552(T) (r > 0.68). Other Lactococcus and Leuconostoc species could be distinguished from the isolates using SDS-PAGE whole-cell protein patterns (r < 0.58) and biochemical characteristics. The 16S rDNA sequencing of four randomly selected strains showed that the strains differed from L. piscium GTC 552(T) by two to three bases in the highly variable region of the sequence. This is the first report on the isolation of L. piscium from vacuum-packaged beef. The gram-positive catalase-negative cocci isolated from vacuum-packaged refrigerated beef have been identified as L. piscium. The findings of this work contribute to the knowledge of the microflora of vacuum-packaged refrigerated beef.

Atopobacter phocae gen. nov., sp. nov., a novel bacterium isolated from common seals.

Two strains of a Gram-positive, catalase-negative, facultatively anaerobic, rod-shaped bacterium isolated from common seals were characterized using phenotypic and molecular taxonomic methods. The two strains closely resembled each other based on their biochemical characteristics, and PAGE analysis of whole-cell protein patterns confirmed their close phenotypic affinity. 16S rRNA gene sequencing showed that the two strains were genetically highly related (99.8% sequence similarity) and that they constitute a new line of descent within the lactic acid group of bacteria. The nearest phylogenetic neighbours of the unknown bacterium were Granulicatella spp., with related taxa such as enterococci, carnobacteria, Desemzia incerta, Lactosphaera pasteurii, Melissococcus plutonius, tetragenococci and vagococci more distantly related. Based on phylogenetic and phenotypic evidence it is proposed that the unknown bacterium from seals be classified in a new genus as Atopobacter phocae gen. nov., sp. nov. The type strain of Atopobacter phocae is CCUG 42358T (= CIP 106392T).

Reclassification of Flavobacterium odoratum (Stutzer 1929) Strains to a New Genus, Myroides, as Myroides odoratus comb. nov. and Myroides odoratimimus sp. nov.

On the basis of genotypic, chemotaxonomic, and phenotypic data, Flavobacterium odoratum was excluded from the emended genus Flavobacterium. In the present study, the known heterogeneity within this species was examined by a polyphasic approach that included DNA-rRNA hybridizations, a determination of DNA base ratios, DNA-DNA hybridizations, a numerical analysis of whole-cell protein patterns, a determination of cellular fatty acid compositions, and a phenotypic analysis. All of the methods revealed the presence of two distinct species. We propose that F. odoratum strains should be placed in a new genus, the genus Myroides, which comprises two species, Myroides odoratus comb. nov. and Myroides odoratimimus sp. nov.

Phenotypic identification and differentiation of Lactococcus strains isolated from animals

Summary Lactococcus lactis subsp. lactis, Lactococcus garvieae and Lactococcus raffinolactis were identified in samples from different domestic animal species, mainly in tonsils of ruminants, dogs and cats, and milk samples from cases of subclinical mastitis in dairy cows. Species differentiation was mainly based on SDS-PAGE analysis of whole-cell protein patterns. A comparison was made with results of biochemical activity tests and sensitivity patterns to clindamycin and oxacillin. These results were used to construct an improved identification scheme for lactococci from animal origin. By SDS-PAGE analysis of whole-cell proteins, Enterococcus seriolicida was shown to be closely related to Lactococcus garvieae .

Taxonomic Analysis of Thermophilic Strains of the Genus Methanobacterium: Reclassification of Methanobacterium thermoalcaliphilum as a Synonym of Methanobacterium thermoautotrophicum

DNA reassociation, a comparative analysis of whole-cell protein patterns, and indirect immunofluorescence methods were used to determine the taxonomic relationships of thermophilic Methanobacterium strains. On the basis of the results dendrograms showing degrees of similarity among the organisms were constructed. The organisms studied are members of three different groups. One group, whose members exhibit 10 to 40% cross-hybridization, includes Methanobacterium wolfeii, “Methanobacterium defluvium” ADZ, and “Methanobacterium thermoflexum” IDZ. The second group contains Methanobacterium thermophilum MT (T = type strain) and Methanobacterium thermoaggregans (levels of DNA relatedness, 30 to 45%). The third group, whose members exhibit 65 to 99% cross-hybridization, includes Methanobacterium thermoautotrophicum ΔHT, F-1, and DV; Methanobacterium thermoformicicum Z-245T; and Methanobacterium thermoalcaliphilum AC60T. The combination of three independent taxonomic methods showed that the group of strains studied is phenotypically, genotypically, and antigenically diverse. The most distinct organisms are M. wolfeii, M. thermoaggregans, M. thermophilum, “M. defluvium,”“M. thermoflexum,” and M. thermoautotrophicum. These species names should be adopted. The results of a DNA-DNA hybridization study (level of hybridization, 99%), an immunological analysis (+3 reaction), and a protein similarity study (level of similarity, 75%) indicate that M. thermoalcaliphilum should be reclassified as a synonym of M. thermoautotrophicum.

Cellular fatty acid compositions of "Achromobacter groups B and E"

Strains of "Achromobacter groups B and E" were examined for cellular fatty acid (CFA) composition to evaluate their chemical relatedness to known bacterial species and groups. The CFAs were liberated from whole cells by base hydrolysis, methylated, and analyzed by capillary gas-liquid chromatography. The CFA profiles of the two groups were identical and were distinct from CFA profiles of all other bacteria we have previously tested. These data provide support for results from whole-cell protein pattern analysis and DNA-DNA and rRNA-DNA hybridization studies, which show that "Achromobacter groups B and E" are biotypes of a single new genus and species.

Polyphasic taxonomic study of the emended genus Arcobacter with Arcobacter butzleri comb. nov. and Arcobacter skirrowii sp. nov., an aerotolerant bacterium isolated from veterinary specimens.

The relationships of 77 aerotolerant Arcobacter strains that were originally identified as Campylobacter cryaerophila (now Arcobacter cryaerophilus [P. Vandamme, E. Falsen, R. Rossau, B. Hoste, P. Segers, R. Tytgat, and J. De Ley, Int. J. Syst. Bacteriol. 41:88-103, 1991]) and 6 reference strains belonging to the taxa Arcobacter nitrofigilis, Arcobacter cryaerophilus, and "Campylobacter butzleri" were studied by using a polyphasic approach, in which we performed DNA-rRNA hybridizations, DNA-DNA hybridizations, a numerical analysis of whole-cell protein patterns after sodium dodecyl sulfate-polyacrylamide gel electrophoresis, an analysis of cellular fatty acid compositions, and a phenotypic analysis and determined DNA base ratios. Our results indicate that "C. butzleri" should be transferred to the genus Arcobacter as Arcobacter butzleri comb. nov., as was suggested by Kiehlbauch and coworkers (J. A. Kiehlbauch, D. J. Brenner, M. A. Nicholson, C. N. Baker, C. M. Patton, A. G. Steigerwalt, and I. K. Wachsmuth, J. Clin. Microbiol. 29:376-385, 1991). A rapid screening of all strains in which we used the sodium dodecyl sulfate-polyacrylamide gel electrophoresis technique revealed five major groups, which were identified by using DNA-DNA hybridization data as A. cryaerophilus (two distinct electrophoretic subgroups), A. butzleri, A. nitrofigilis, and a new species, for which we propose the name Arcobacter skirrowii. The phylogenetic position within rRNA superfamily VI was established for each species. A. butzleri strains and strains belonging to one of the electrophoretic subgroups of A. cryaerophilus had similar fatty acid contents. An analysis of fatty acid compositions allowed clear-cut differentiation of all of the other groups. All of the species could be distinguished by using classical phenotypic tests, although erroneous identifications due to a shortage of clear-cut differentiating tests could occur.

Rapid identification of environmental isolates of Pseudomonas aeruginosa, P. fluorescens and P. putida by SDS-PAGE analysis of whole-cell protein patterns

Environmental isolates of fluorescent pseudomonads grown to early stationary phase in glucose-enriched Luria broth were treated with proteinase K in sodium dodecylsulphate (SDS) lysis buffer and subsequently analyzed by polyacrylamide gel electrophoresis (PAGE). Four silver-staining protein-fragment bands could be used for rapid identification at the species level. Pseudomonas aeruginosa isolates were easily recognized by a unique banding pattern. Isolates considered to be P. fluorescen from biochemical and physiological tests (classical biotypes I, II, III, IV and V) also had a characteristic banding pattern, which in turn was different from that of P. putida isolates (classical biotype A). A residual group representing intermediate isolates of P. fluorescens (new biotype VI of Barrett et al., J. Gen. Microbiol. 132, 1986) or P. putida (biotype B) had a banding pattern similar to that of classical P. fluorescens biotypes. On the other hand, a group representing other intermediate isolates of P. putida (new biotype C of Barrett et al., J. Gen. Microbiol. 132, 1986) had a unique banding pattern resembling that of classical P. putida biotype A. A small number of protein fragment bands appearing in SDS-PAGE analysis of whole-cell lysates seems adequate for a rapid identification at the species level of P. aeruginosa, P. fluorescens and P. putida isolated from natural environments.

Rapid identification of environmental isolates of Pseudomonas aeruginosa, P. fluorescens and P. putida by SDS-PAGE analysis of whole-cell protein patterns

Rapid identification of environmental isolates of Pseudomonas aeruginosa, P. fluorescens and P. putida by SDS-PAGE analysis of whole-cell protein patterns

Rapid identification of environmental isolates of Pseudomonas aeruginosa, P. fluorescens and P. putida by SDS-PAGE analysis of whole-cell protein patterns

Rapid identification of environmental isolates of Pseudomonas aeruginosa, P. fluorescens and P. putida by SDS-PAGE analysis of whole-cell protein patterns