Taxonomic studies of some cultured strains of Cyanobacteria (Nostocales) isolated from the Yenisei River basin

Twenty-one strains of cyanobacteria representing the genera Anabaena, Cylindrospermum, Mojavia, Nostoc, Trichormus, and Wollea (Nostocales, Cyanobacteria) isolated from algocenoses of the Yenisei River basin (eastern Siberia, Russia) were taxonomically studied. New taxa characteristic of this region were discovered. The properties of pure cultures of Anabaena sedovii, A. zinserlingii,Cylindrospermum stagnale f. tortuosum, Nostoc kihlmani, Trichormus variabilis f. tenuis, and Wollea saccata have never been described before. The primary taxonomic features (the position of akinetes and heterocysts, the width of vegetative cells, akinetes and heterocysts, and the shape of terminal cells) of these cyanobacteria when cultured were shown to fit the diagnosis of the identified taxa.     

Taxonomic diversity of Nitrosovibrio strains isolated from building sandstones

Abstract Five strains of Nitrosovibrio , isolated from building sandstones of historic monuments in Germany, were compared. They were characterized by the presence or absence of carboxysomes, generation time, tolerance to ammonium chloride and sodium chloride, and temperature optima. Three strains were distinguishable by DNA homologies, ultrastructure, and growth mode. Only two strains were closely related, indicated e.g. by a DNA homology of 72%. Taxonomical diversity of the endolithic Nitrosovibrio strains may be related to the existence of microniches in building sandstones. Cell aggregation was observed for endolithic cells and for cells of isolated Nitrosovibrio strains. This could be a response to acidic environments.     

Taxonomic studies on Cryptococci and related yeasts

A taxonomic methodology based on current keys and the ready availability of refrigerated slant media is presented. It utilizes defined media, surface inoculation, and reagent indicators which simplify final readings. Yeast Nitrogen Base w/o Amino Acids or Ammonium Sulfate (DIFCO) is used as a source of vitamins and trace nutrilites. The medium consists of the Seitz-filtered vitamin base (0.1 % w/v), nitrogen source (0.5 %), and carbon source (1.0 %) mixed into 1.5 % agar slants. Test inocula are taken from a vitamin base-ammonium sulfate-dextrose positive control slant. Cultures are incubated for 2–4 days at 25° C. Nitrate assimilation is tested with the usual sulfanilic acid-alphanaphthylamine reagents. Carbon assimilation is tested with 0.04 % aqueous brom-phenol-blue. These reagent tests help discriminate between negative, weak, and doubtful test results. Tables of test results onCryptococcus, Rhodotorula, and non-fermentingTorulopsis species are presented.

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Zusammenfassung Eine taxonomische, an gegenwärtige Schlüssel gebundene Methode und die leichte Verfügbarkeit eines gekühlten, schrägen Nährbodens werden beschrieben. Es sind definierte Nährböden, Oberflächenverimpfung, und Reagensindikatoren benützt, um die Endablesung zu erleichtern. Hefen-Stickstoffbase mit Aminosäuren oder Ammonium sulfate (DIFCO) war als Vitaminquelle und als Spurnährstoffe benützt. Der Nährboden besteht aus einer Seitz-filtrierten Vitaminbase (0.1 % w/o), Stickstoffquelle (0.5 %) und Kohlenstoffquelle (1.0 %) mit einem 1.5 % Schrägagar. Testinoculums sind von einem positiven Kontrol-schrägagar mit Vitaminbase-Ammonium sulfate-Dextrose genommen. Kulturen sind für 2 bis 4 Tage bei 25° C bebrühtet. Nitrat-Assimilation ist mit dem üblichen sulfanilic acid-alphanaphtylamine Reagens getestet. Kohlenstoff-Assimilation ist mit 0.04 % wässerigem Bromphenol-blue getestet. Diese Reagenstests erleichtern die Unterscheidung unter den negativen, schwachen und zweifelhaften Testergebnissen. Testergebnisse anCryptocoscus, Rhodotorula und an nicht vergärendenTorulopsis-Arten sind beschrieben.

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Presented at the American Society for Microbiology meeting in Miami Beach on May 5, 1969.     

Taxonomic and phylogenetic evaluation of Limnothrix strains (Oscillatoriales, Cyanobacteria) by adding Limnothrix planktonica strains isolated from central China

Six Limnothrix strains, isolated for the first time from a shallow eutrophic lake in central China, were taxonomically and phylogenetically evaluated by investigating their polyphasic characteristics, including morphological features, cellular ultrastructures, and 16S rRNA gene sequences. All the six strains were morphologically similar, and their trichomes were in average 1.7 μm wide and cells 4.0 μm long, and having small gas vesicles within cells, and therefore identified as Limnothrix planctonica (Woloszynska) Meffert. Cellular ultrastructures of them showed that peripheral thylakoids with 3–5 parallel layers were parietally distributed in the cells. The phylogenetic results based on the 16S rRNA gene sequences showed that all the Limnothrix strains, including the six in this study and those from the Genbank, formed two distinct clusters. The similarity in 16S rDNA sequences between these two clusters was lower than 90%, indicating that these Limnothrix strains belong to different genera. This is the first report on the morphology and phylogeny of L. planctonica strains, providing the new information on taxonomy of the genus Limnothrix.     

Analysis of Salmonella enteritidis strains isolated from food-poisoning cases in Taiwan by pulsed field gel electrophoresis, plasmid profile and phage typing

AIMS: To establish the molecular typing data for Salmonella enteritidis due to its increasing role in Salmonella infections in Taiwan. METHODS AND RESULTS: Sixty-three Salm. enteritidis strains isolated from related and unrelated patients suffering from food-borne poisoning during 1991-97 were collected and subjected to pulsed field gel electrophoresis (PFGE), plasmid analysis and phage typing. For PFGE, XbaI, SpeI and NotI restriction enzymes were used for chromosomal DNA digestion. The results showed that, for these 63 Salmonella strains, 10 PFGE pattern combinations were found. Of these, pattern X3 S3 N3 was the major subtype, since 46 strains isolated from different locations at different times during 1991-97 showed this PFGE pattern. Plasmid analysis showed only three plasmid profiles and phage typing showed that most of the Salmonella strains were of the phage type PT4. CONCLUSION: Most of the Salm. enteritidis strains circulating in Taiwan are of very similar genetic types or are highly related and that strains of PFGE pattern X3 S3 N3 are the prevalent and recirculating strains of Salm. enteritidis which caused food-poisoning cases in Taiwan in 1991-97. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information that in Salmonella infection, certain subtypes of Salm. enteritidis should be scrutinized.     

Taxonomic studies on Corynebacterium beticola (Abdou)

Morphological, physiological and chemical studies were performed on the type strain of Corynebacterium beticola in an attempt to clarify its taxonomy. The results indicate that Corynebacterium beticola is a Gram negative, facultatively anaerobic bacterium which should be assigned to the genus Erwinia as Erwinia herbicola .     

Characterization of hydrocarbon-degrading bacterial strains isolated from oil-polluted soil

Using enrichment culturing method, a microbial population was detected in an oil-contaminated soil nearby an extraction field. Isolated strains were able use medium-length n-alkanes as sole carbon and energy source as assessed by growth experiments. Results showed a high diversity among strains at molecular level, and also in the metabolic profiles. Physiological and biochemical tests showed a similarity within a group of four strains, as confirmed by Biolog MicroLog analysis. Based on 16S rDNA sequences strains were identified as Rhodococcus erythropolis, Acinetobacter baumanii, Burkholderia cepacia and Achromobacter xylosoxidans. Alkane monooxygenase gene (alkB) was successfully detected in all our strains and for the first time alkB genes were found in an A. xylosoxidans strain. This bacterial species has been previously reported as part of microbial communities from oil polluted environments, but there are few studies that address mechanisms details of A. xylosoxidans involvement in n-alkane degradation process.     

Equine and canine Anaplasma phagocytophilum strains isolated on the island of Sardinia (Italy) are phylogenetically related to pathogenic strains from the United States

The presence of Anaplasma phagocytophilum, a tick-transmitted zoonotic pathogen, was investigated in Sardinia using a molecular approach. Phylogenetic analysis revealed that Sardinian strains are genetically distinct from the two lineages previously described in Europe and are closely related to strains isolated in different areas of the United States.     

Biodegradation of chrysene by the bacterial strains isolated from oily sludge

The biodegradation studies were conducted to test the ability of the bacterial strains (Chry2 and Chry3) isolated from the oily sludge obtained from Gujarat refinery, India, for utilization of chrysene in the liquid medium. Biodegradation of the compound was confirmed using gas chromatography and the percent degradation was calculated to be 15.0 and 17% by Chry2 and Chry3, respectively. The biodegradation results were supported by increase in viable cell count and dry biomass, in the presence of chrysene as the sole carbon source. Both the cultures produced biosurfactant which was indicated by the reduction in surface tension of the growth medium. Presence of catechol 2, 3-dioxygenase gene in Chry3 indicated its potential for degradation of PAHs through meta cleavage degradation pathway. Both the strains were found to possess catechol 1,2-dioxygenase and catechol 2,3-dioxygenase enzyme activities. Based on morphological and biochemical tests, the cultures were tentatively identified as Bacillus sp. (Chry2) and Pseudomonas sp. (Chry3).     

Enterotoxigenicity of Staphylococcus aureus strains isolated from poultry: raw poultry carcases as a potential food-poisoning hazard

Staphylococcus aureus strains were isolated from end-of-lay poultry carcases obtained from a plant at two different stages of processing before and after storage at different temperatures. These strains were supplemented with Staph. aureus strains isolated from poultry from a wide range of sources and biotyped, phage typed, and tested for production of enterotoxins A-E. The isolates were found to consist of poultry and human specific strains and each of these groups contained strains able to produce enterotoxin. Poultry strains produced only enterotoxin D whereas human strains produced enterotoxins A, C and D. The hen carcases used in storage experiments were found to be naturally contaminated with enterotoxin D producing staphylococci. No enterotoxin D could be detected on any of the carcases even after storage at temperatures which allowed multiplication of the organisms to occur (final Staph. aureus counts ranged from 10² to 10⁷/16 cm² of breast skin).     

Characterization of "Pasteurella" piscicida isolated from white perch and cultivated yellowtail.

The morphological, physiological, and biochemical characteristics of twelve "Pasteurella" piscicida strains isolated from white perch and yellowtail are described and the present uncertain taxonomic status of the organisms is discussed. The organisms isolated were gram-negative rods showing bipolar staining and pleomorphism. No spores or flagella were observed. They were non motile and viscid colonies were formed. Growth was observed in a temperature range of 20 to 30 C and the salinity range of growth was between 0.5% and 4.0%. They were aerobic and facultative anaerobic. Oxidase and catalase were produced. Nitrates were not reduced to nitrites. Lysine and ornithine decarboxylases were not produced but arginine dihydrolase was produced. The egg yolk reaction was positive. Tween 80 and tributyrin were decomposed. Phosphatase was produced. Beta hemolysis was revealed on a medium containing defibrinated blood from chickens or carp but not from mammals. Methyl red and Voges-Proskauer tests were positive, and acetoin was produced from 2,3-butanediol. Glucose was fermented without gas production. Acid was produced from fructose, mannose, galactose, sucrose, maltose, dextrin and glycerol. These organisms differed from all other members of the genus Pasteurella with respect to nitrate reduction, arginine dihydrolase, Voges-Proskauer and methyl red tests. The formation of viscid colonies and inability to grow in a medium without sodium chloride or at 37 C were additional characteristics of these organisms.     

Utilization of drought-tolerant bacterial strains isolated from harsh soils as a plant growth-promoting rhizobacteria (PGPR)

Drought stress adversely affects plant health and productivity. Recently, drought-resistant bacterial isolates are used to combat drought resistance in crops. In this in vitro study, 20 bacterial isolates were isolated from harsh soil; their drought tolerance was evaluated using four concentrations of polyethylene glycol (PEG) 6000. The two most efficient isolates (DS4 and DS9) were selected and identified using 16S rRNA genetic sequencing. They were registered in the NCBI database and deposited under accession numbers MW916285 and MW916307 for Bacillus cereus (DS4) and Bacillus albus (DS9), respectively. These isolates were screened for plant growth-promoting properties compared to non-stressed conditions. Biochemical parameters; Proline, salicylic acid, gibberellic acid (GA), indole acetic acid (IAA), antioxidant activity, and antioxidant enzymes were measured under the same conditions, and in vitro seed germination was tested under stress conditions and inoculation with selected isolates. The results showed that under the harsh conditions of PEG6000, DS4 produced the highest amount of IAA of 1.61 µg/ml, followed by DS9 with 0.9 µg/ml. The highest amount of GA (49.95 µg/ml) was produced by DS9. On the other hand, the highest amount of siderophore was produced from DS4 isolate followed by DS9. Additionally, DS4 isolate recorded the highest exopolysaccharide (EPS) content of 3.4 mg/ml under PEG (-1.2 MPa) followed by DS9. The antioxidant activity increased in PEG concentrations depending manner, and the activity of the antioxidant enzymes increased, as catalase (CAT) recorded the highest activity in DS4 with an amount of 1.095 mg/ml. additionally, an increase in biofilm formation was observed under drought conditions. The isolated mixture protected the plant from the harmful effects of drought and showed an increase in the measured variables. Under unstressed conditions, the highest rates of emulsification index (EI 24%) were obtained for DS4 and DS9, at 14.92 and 11.54, respectively, and decreased under stress. The highest values of germination, total seedling length, and vigor index were obtained upon inoculation with the combination of two strains, and were 100%, 4.10 cm, and 410, respectively. Therefore, two strains combination is an effective vaccine capable of developing and improving drought tolerance in dryland plants.     

Taxonomic studies on Solarium section Solarium (Maurella)

Various taxonomic methods are being used to clarify the systematics of Solarium section Solanum , the large and variable species group centring around the black nightshade, S. nigrum. To put the experimental work into perspective, a summary of former taxonomic treatments is given, together with an outline of the provisional taxonomic framework of the section, initially established from earlier classical studies, and supplemented by later experimental work.
Experimentation with living populations has shown that Solanum taxonomy is complicated by several factors. In addition to the historical factors, phenotypic plasticity, genetic variation, the existence of a polyploid series, and the possibilities of interspecific hybridization accompanied by various pre- and post-zygotic isolating mechanisms are dealt with.
The various taxonomic categories attached to Solanum accessions during earlier classical studies derive support from the crossability behaviour of the taxa concerned. Morphological divergence is generally accompanied by genetical isolation which can manifest itself at any time between pollination and the maturation of the F₂ progeny. Data from the hybridization studies, in which 14 diploid species, five tetraploid species, four hexaploid species and one octoploid taxon were used, are presented in detail.     

Physiological traits of bacterial strains isolated from phenol-degrading aerobic granules

The physiological characteristics of ten bacterial strains isolated from phenol-degrading aerobic granules were evaluated in order to identify competitive traits for dominant growth in aerobic granules. The ten strains showed a wide diversity in specific growth rates and oxygen utilization kinetics, and could be divided into four catabolic types of phenol degradation. While some strains degraded phenol mainly via the meta pathway or the ortho pathway, other strains degraded phenol via both these pathways. The ten strains also exhibited high levels of autoaggregation and coaggregation activity. Within the collection of ten strains, 36.7% of all possible strain pairings displayed a measurable degree of coaggregation. Strain PG-08 possessed the strongest autoaggregation activity and showed significant coaggregation (coaggregation indices of 67% to 74%) with PG-02. The three strains PG-01, PG-02 and PG-08 belonging to dominant groups in the granules possessed different competitive characteristics. Microcosm experiments showed the three strains could not coexist at the high phenol concentration of 250 mg L(-1), but could coexist at lower phenol concentrations in a spatially heterogeneous environment. This study illustrated that the spatial heterogeneity provided by the aerobic granules led to niche differentiation and increased physiological diversity in the resident microbial community.     

Phenotypic, antigenic, and molecular characterization of Pasteurella piscicida strains isolated from fish.

We compared Pasteurella piscicida strains isolated from different fish species in several European countries with strains isolated in Japan and the United States. The taxonomic analysis revealed that, regardless of the geographic origin and source of isolation, all the strains exhibited the same biochemical and physiological characteristics. Serological assays with different rabbit antisera demonstrated a high level of antigenic similarity among strains, with cross-agglutination titers of 20,480 to 40,960. This serological homogeneity was supported by the lipopolysaccharide (LPS) and membrane protein profiles. All the P. piscicida strains had the same electrophoretic LPS pattern, showing O side chains with a ladder-like structure, and shared at least four major outer membrane proteins, of 20, 30, 42, and 53 kDa. Western blot (immunoblot) analysis with LPS and protein indicated that all the P. piscicida strains are immunologically related. In addition, the chromosomal DNA fingerprint patterns obtained for the European strains with the enzymes EcoRI and BamHI were practically identical to those of the Japanese and U.S. strains. Although some differences were found in the plasmid profiles of P. piscicida, a large number of strains possessed in common plasmid bands of 20 and 7 MDa. In addition, a plasmid of 50 MDa was present in the majority of the European strains. Restriction endonuclease analysis demonstrated the genetic homology of the plasmid bands shared by most of the European strains. All the P. piscicida strains had the same drug resistance patterns, indicating that a correlation between plasmid carriage and resistance to a specific antimicrobial agent cannot be established. The high levels of phenotypic, serological, and genetic homogeneity found among the P. piscicida strains should facilitate the development of DNA probes with diagnostic purposes as well as the design of effective vaccines.     

Phenotypic, antigenic, and molecular characterization of Pasteurella piscicida strains isolated from fish.

We compared Pasteurella piscicida strains isolated from different fish species in several European countries with strains isolated in Japan and the United States. The taxonomic analysis revealed that, regardless of the geographic origin and source of isolation, all the strains exhibited the same biochemical and physiological characteristics. Serological assays with different rabbit antisera demonstrated a high level of antigenic similarity among strains, with cross-agglutination titers of 20,480 to 40,960. This serological homogeneity was supported by the lipopolysaccharide (LPS) and membrane protein profiles. All the P. piscicida strains had the same electrophoretic LPS pattern, showing O side chains with a ladder-like structure, and shared at least four major outer membrane proteins, of 20, 30, 42, and 53 kDa. Western blot (immunoblot) analysis with LPS and protein indicated that all the P. piscicida strains are immunologically related. In addition, the chromosomal DNA fingerprint patterns obtained for the European strains with the enzymes EcoRI and BamHI were practically identical to those of the Japanese and U.S. strains. Although some differences were found in the plasmid profiles of P. piscicida, a large number of strains possessed in common plasmid bands of 20 and 7 MDa. In addition, a plasmid of 50 MDa was present in the majority of the European strains. Restriction endonuclease analysis demonstrated the genetic homology of the plasmid bands shared by most of the European strains. All the P. piscicida strains had the same drug resistance patterns, indicating that a correlation between plasmid carriage and resistance to a specific antimicrobial agent cannot be established. The high levels of phenotypic, serological, and genetic homogeneity found among the P. piscicida strains should facilitate the development of DNA probes with diagnostic purposes as well as the design of effective vaccines.