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Interaction of the bacteria Xenorhabdus nematophila (Enterobactericeae) and Bacillus subtilis (Bacillaceae) with the hemocytes of larval Malacosoma disstria (Insecta: Lepidoptera: Lasiocampidae).

Malacosoma disstria larvae are a pest of deciduous trees. Little is known on the interaction of bacteria with the immediate hemocytic antimicrobial responses of these insects. Incubating dead Xenorhabdus nematophila and Bacillus subtilis with a mixture of serum-free granular cells and plasmatocytes in vitro revealed differential bacterial-hemocyte adhesion and differential discharge of lysozyme and phenoloxidase but not total protein. Although active phenoloxidase adhered equally to both bacterial species, X. nematophila limited enzyme activation whereas B. subtilis enhanced activation. Serum with active phenoloxidase (as opposed to tropolone-inhibited phenoloxidase) and purified insect lysozyme increased bacterial-hemocyte adhesion of both bacterial species. An apolipophorin-III-like protein when incubated with hemocytes, limited their responses to glass slides and bacterial adhesion. However, initial binding of the protein to both bacteria increased granular cell levels with bacteria while lowering the plasmatocyte levels with adhering procaryotes. The protein also increased lysozyme and phenoloxidase activities. Although B. subtilis in vivo elicited a nodulation-based decline in total hemocyte counts and did not affect hemocyte viability, dead X. nematophila elevated hemocyte counts and damaged the hemocytes as lipopolysaccharide levels increased and X. nematophila emerged into the hemolymph. Apolipophorin-III-like protein once bound to the bacteria slowed their removal from the hemolymph.

Animals↗

[The importance of aerobic spore-forming Bacillaceae in routine demonstration of microorganisms (author's transl)].

According to our studies the number of spore-forming bacilli at the average amounts to 2 to 15% within the air and food flora. Due to their presence in the air they can contaminate test samples. Likewise it is to be taken into consideration that they can cause pathological changes in susceptible organisms. In our laboratory we isolated aerobic bacilli from biological material in seven cases whereby pathogenicity was established in five cases thereof.

Aerobiosis↗

Viability and stability of biological control agents on cotton and snap bean seeds.

Cotton and snap bean were selected for a multi-year, multi-state regional (south-eastern USA) research project to evaluate the efficacy of both commercial and experimental bacterial and fungal biological control agents for the management of damping-off diseases. The goal for this portion of the project was to determine the viability and stability of biological agents after application to seed. The biological seed treatments used included: (1) Bacillaceae bacteria, (2) non-Bacillaceae bacteria, (3) the fungus Trichoderma and (4) the fungus Beauveria bassiana. Seed assays were conducted to evaluate the following application factors: short-term (< or = 3 months) stability after seed treatment; quality (i.e. isolate purity); compatibility with chemical pesticides and other biocontrol agents; application uniformity between years and plant species. For the bacterial treatments, the Bacillaceae genera (Bacillus and Paenibacillus) maintained the greatest population of bacteria per seed, the best viability over time and the best application uniformity across years and seed type. The non-Bacillaceae genera Burkholderia and Pseudomonas had the least viability and uniformity. Although Beauveria bassiana was only evaluated one year, the seed fungal populations were high and uniform. The seed fungal populations and uniformity for the Trichoderma isolates were more variable, except for the commercial product T-22. However, this product was contaminated with a Streptomyces isolate in both the years that it was evaluated. The study demonstrated that Bacillaceae can be mixed with Trichoderma isolates or with numerous pesticides to provide an integrated pest control/growth enhancement package.

Bacillaceae↗

Growth and survival of immature Heamatobia irritans (Diptera; Muscidae) is influenced by bacterial isolated from cattle manure and conspecific larvae.

Twenty species of bacteria were isolated from cattle manure and seven species were isolated from the gut of larval horn fly Hematobia irritans (L.). Bacteria in manure belonged to the Bacillaceae, Pseudomonadaceae, Micrococcaceae, Corynebacteriaceae, Enterobacteriaceae, Microbacteriaceae, and two unassigned genera. Gut bacteria belonged to the Enterobacteriaceae, Bacillaceae, Neisseriaceae, and Pseudomonadaceae. H. irritans larval survival and growth on the various bacterial species were evaluated by rearing larvae in sterilized cattle manure that was inoculated with single bacterial isolates. H. irritans larvae failed to develop in sterilized, uninoculated manure, indicating that bacteria are necessary for larval development. Survival averaged 74% in nonsterilized manure and ranged from 4 to 53% in manure with individual isolates. Survival was highest when larvae were reared on manure inoculated with Pseudomonadaceae, Corynebacteriaceae, Micrococcaceae, and Bacillaceae and was lowest when reared in manure inoculated with Enterobacteriaceae and Microbacteriaceae. Pupal weights were heaviest when reared on the Flavobacteria, followed by the Pseudomonadaceae and Corynebacteriaceae. Pupae averaged 4.9 +/- 0.08 mg when reared on gram-negative isolates, compared with 3.6 +/- 0.09 mg when reared on gram-positive isolates. Pupal weights were not significantly correlated with larval survival, indicating that bacteria that promote growth do not necessarily promote survival. A reproductive index was used as a measure of fitness and was highest for larvae reared in the nonsterile control, followed most closely by Pseudomonadaceae and Corynebacteriaceae. These groups appeared to best meet the nutritional requirements of larvae and may be used in further experiments to define an artificial rearing media for H. irritans.

Actinomycetales↗

Thermalkalibacillus uzonensis gen. nov. sp. nov, a novel aerobic alkali-tolerant thermophilic bacterium isolated from a hot spring in Uzon Caldera, Kamchatka.

A novel thermophilic, alkali-tolerant, and CO-tolerant strain JW/WZ-YB58(T) was isolated from green mat samples obtained from the Zarvarzin II hot spring in the Uzon Caldera, Kamchatka (Far East Russia). Cells were Gram-type and Gram stain-positive, strictly aerobic, 0.7-0.8 mum in width and 5.5-12 mum in length and produced terminal spherical spores of 1.2-1.6 mum in diameter with the mother cell swelling around 2 mum in diameter (drumstick-type morphology). Cells grew optimally at pH(25 degrees C) 8.2-8.4 and temperature 50-52 degrees C and tolerated maximally 6% (w/v) NaCl. They were strict heterotrophs and could not use either CO or CO(2 )(both with or without H(2)) as sole carbon source, but tolerated up to 90% (v/v) CO in the headspace. The isolate grew on various complex substrates such as yeast extract, on carbohydrates, and organic acids, which included starch, D: -galactose, D: -mannose, glutamate, fumarate and acetate. Catalase reaction was negative. The membrane polar lipids were dominated by branched saturated fatty acids, which included iso-15:0 (24.5%), anteiso-15:0 (18.3%), iso-16:0 (9.9%), iso-17:0 (17.5%) and anteiso-17:0 (9.7%) as major constituents. The DNA G+C content of the strain is 45 mol%. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain JW/WZ-YB58(T) is distantly (<93% similarity) related to members of Bacillaceae. On the basis of 16S rRNA gene sequence, physiological and phenotypic characteristics, the isolate JW/WZ-YB58(T) (ATCC BAA-1258; DSM 17740) is proposed to be the type strain for the type species of the new taxa within the family Bacillaceae, Thermalkalibacillus uzoniensis gen. nov. sp. nov. The Genbank accession number for the 16S rRNA gene sequence is DQ221694.

Bacteria, Aerobic↗

Phylogenetic relationships between Bacillus species and related genera inferred from comparison of 3' end 16S rDNA and 5' end 16S-23S ITS nucleotide sequences.

The nucleotide sequences of the 3' end of the 16S rDNA and the 16S-23S internal transcribed spacer (ITS) of 40 Bacillaceae species were determined. These included 21 Bacillus, 9 Paenibacillus, 6 Brevibacillus, 2 Geobacillus, 1 Marinibacillus and 1 Virgibacillus species. Comparative sequence analysis of a 220 bp region covering a highly conserved 150 bp sequence located at the 3' end of the 16S rRNA coding region and a conserved 70 bp sequence located at the 5' end of the 16S-23S ITS of the 40 species and six sequences available in GenBank were used to infer the phylogenetic relationships between all 46 taxa. When a maximal distance (D(max), where D refers to the number of nucleotide substitutions per site) of 0.31 was introduced as a threshold to determine groupings, 10 phylogenetically distinct clusters were revealed. Twenty-six Bacillus species were separated in seven groups (I, II, III, IV, V, VI and X), but Bacillus circulans remained ungrouped. All six Brevibacillus species under study were in Group VII. The nine Paenibacillus species fell into two distinct groups (VIII and IX). Species with D(max) values within 0.05 were considered to be very closely related. These were Bacillus psychrophilus and Bacillus psychrosaccharolyticus in Group II; 'Bacillus maroccanus' and Bacillus simplex in Group II; Bacillus amyloliquefaciens, Bacillus atrophaeus, Bacillus mojavensis and Bacillus subtilis in Group VI; Bacillus fusiformis and Bacillus sphaericus in Group VI; Brevibacillus brevis and Brevibacillus formosus in Group VII; Paenibacillus gordonae and Paenibacillus validus in Group VIII; and Bacillus anthracis, Bacillus cereus, Bacillus mycoides and Bacillus thuringiensis in Group X. The phylogenetic classification presented here is, in general, in agreement with current classifications based on phenotypic and molecular data. Our findings suggest, however, that in some cases, further divisions or, conversely, further groupings might be warranted. Should current classifications be re-examined in the light of our results, D(max) values of 0.31 and 0.05, as exemplified here, may prove useful threshold values for the grouping of Bacillaceae into taxa akin to genera and species, respectively. These D(max) thresholds may also reveal, in a different way, bacterial species for which further characterization might be warranted for proper classification and/or reassignment.

3' Untranslated Regions↗

Enumeration and characterization of cellulolytic bacteria from refuse of a landfill.

Enumeration and phenotypic characterization of aerobic cellulolytic bacteria were performed on fresh, 1 year old and 5 years old refuse samples of a French landfill site. Numbers of cellulolytic bacteria ranged from 1.1x10(6) to 2.3x10(8) c.f.u. (g dry wt.)(-1) and were lower in 5 years old refuse samples. A numerical analysis of phenotypic data based on 80 biochemical tests and performed on 321 Gram-positive isolates from refuse, revealed a high phenotypic diversity of cellulolytic bacteria which were distributed into 21 clusters. Based on the phenotypic analysis and the sequencing of 16S rDNA of five representative strains of major clusters, the predominant cellulolytic groups could be assigned to the family of Bacillaceae and to the genera Cellulomonas, Microbacterium and Lactobacillus. Furthermore, chemical parameters such as pH, carbohydrates and volatile solid contents influenced the composition of the cellulolytic bacterial groups which were reduced essentially to the family of Bacillaceae in the oldest refuse samples.

Journal Article↗

Comparative allostery of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthetase as a molecular basis for classification.

The allosteric pattern of control for 3-deoxy-d-arabino-heptulosonate 7-phosphate (DAHP) synthetase has previously been shown to be strongly conserved among the member species of a given genus in bacteria. The implications of this finding as a procedural tool of bacterial phylogeny were pursued by a study of two organisms, Sporosarcina ureae and Aeromonas formicans, the taxonomic positions of which have been historically controversial. S. ureae has characteristics of both Bacillaceae and Micrococcaceae, and A. formicans has characteristics of both Enterobacteriaceae and pseudomonads. Since the patterns of control for DAHP synthetase in all four of these microbial groups are different from one another but internally homogeneous within each group, the results obtained from the two test organisms were unambiguous. It was concluded that S. ureae is properly classified within Bacillaceae, probably deserving generic rank, and that A. formicans belongs with the family Enterobacteriaceae.

Aeromonas↗

H2-producing bacterial communities from a heat-treated soil inoculum.

Hydrogen gas (approximately 60% H(2)) was produced in a continuous flow bioreactor inoculated with heat-treated soil, and fed synthetic wastewater containing glucose (9.5 g l(-1)). The pH in the bioreactor was maintained at 5.5 to inhibit consumption of H(2) by methanogens. The objective of this study was to characterize bacterial communities in the reactor operated under two different hydraulic retention times (HRTs of 30-h and 10-h) and temperatures (30 degrees C and 37 degrees C). At 30-h HRT, the H(2) production rate was 80 ml h(-1) and yield was 0.91 mol H(2)/mol glucose. At 10-h HRT, the H(2) production rate was more than 5 times higher at 436 ml h(-1), and yield was 1.61 mol H(2)/mol glucose. Samples were removed from the reactor under steady-state conditions for PCR-based detection of bacterial populations by ribosomal intergenic spacer analysis (RISA). Populations detected at 30-h HRT were more diverse than at 10-h HRT and included representatives of Bacillaceae, Clostridiaceae, and Enterobacteriaceae. At 10-h HRT, only Clostridiaceae were detected. When the temperature of the 10-h HRT reactor was increased from 30 degrees C to 37 degrees C, the steady-state H(2) production rate increased slightly to 463 ml h(-1) and yield was 1.8 mol H(2)/mol glucose. Compared to 30 degrees C, RISA fingerprints at 37 degrees C from the 10-h HRT bioreactor exhibited a clear shift from populations related to Clostridium acidisoli (subcluster Ic) to populations related to Clostridium acetobutylicum (subcluster Ib).

Bacillaceae↗

Investigation of the hygienic safety of continuous positive airways pressure devices after reprocessing.

With the widespread use of continuous positive airways pressure (CPAP) therapy, the safety of CPAP devices after reprocessing is the subject of debate. In this study, the contamination of CPAP devices and the effectiveness of disinfection was investigated. A total of 122 CPAP devices were examined including 50 CPAP devices used by patients, which were examined before and after reprocessing. Seventy-two new CPAP devices that had not been in contact with patients served as controls. The results of this study show that the microbial contamination of new and used CPAP devices is only minimal. Contaminating micro-organisms were predominantly micro-organisms reflecting the normal environmental microflora such as Penicillium spp., Aspergillus spp., Micrococcaceae and Bacillaceae. Gram-negative species could only be found in rare cases. The data obtained give no indication of poor disinfection of CPAP devices.

Air Microbiology↗

Microbial community analysis of thermophilic contact oxidation process by using ribosomal RNA approaches and the quinone profile method.

Microbial community structure of a lab scale thermophilic aerobic wastewater treatment reactor was analyzed by a combination of culture-independent methods. Quinone profile method provides for chemical analysis of respiratory quinone molecular species, which corresponds to bacterial groups. Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rDNA partial sequences (PCR-DGGE) clarifies community changes at species level, as DGGE can separate DNA fragments of different sequences. Certain phvlogenetic groups of bacterial cells can be labeled by fluorescence in situ hybridization (FISH). Quinone profile showed a predominant presence of MK-7. PCR-DGGE revealed that constituents of the community were unchanged during the stable phase. FISH demonstrated the existence of the relatives of Bacillus lentus and B. thermocloacae in considerable proportions. The community was mainly composed of Bacillaceae, and obligate thermophilic and mesophilic Bacillus appeared in spite of the temperature fluctuation from 35 degrees C to 60 degrees C. The combination of these culture-independent methods revealed the community precisely enough to evaluate the reactor performance.

Bacillaceae↗

Suprageneric classification of thermoactinomyces vulgaris by nucleotide sequencing of 5S ribosomal RNA.

The 5S rRNA nucleotide sequence of Thermoactinomyces vulgaris was determined and compared with published sequences of representative Gram-positive bacteria. The primary and secondary structure of the sequence is of the type characteristic of Gram-positive bacteria that have DNA with a low proportion of guanine plus cytosine. It was evident from the phylogenetic trees that T. vulgaris has little in common with actinomycetes but is related to the genus Bacillus, showing a moderately high relationship with B. stearothermophilus. The taxonomic implications of these relationships are discussed and an emended description of the family Bacillaceae is given.

Bacillaceae↗

Tenuibacillus multivorans gen. nov., sp. nov., a moderately halophilic bacterium isolated from saline soil in Xin-Jiang, China.

Two Gram-positive, rod-shaped, spore-forming and moderately halophilic bacteria (strains 28-1T, 28-4), isolated from a soil sample from a neutral salt lake in Xin-Jiang, China, were characterized polyphasically. On the basis of fasta (ungapped) analyses of 16S rRNA gene sequences, strains 28-1T and 28-4 were shown to belong to the Bacillaceae and to be closely related to Filobacillus milensis DSM 13259T (97.0 %) and Bacillus haloalkaliphilus DSM 5271T (95.7 %). 16S rRNA gene sequence similarity with other recognized species was not more than 94.1 %. Phylogenetic, chemotaxonomic, physiological and biochemical data supported the differentiation of these novel strains from F. milensis and B. haloalkaliphilus. Therefore these two previously unidentified strains are considered to represent a new genus and species, for which the name Tenuibacillus multivorans gen. nov., sp. nov. is proposed. The type strain is 28-1T (=AS 1.3442T=NBRC 100370T).

Bacillaceae↗

Salinibacillus aidingensis gen. nov., sp. nov. and Salinibacillus kushneri sp. nov., moderately halophilic bacteria isolated from a neutral saline lake in Xin-Jiang, China.

Three Gram-positive, moderately halophilic, heterotrophic bacterial strains were isolated from a neutral saline lake in the Xin-Jiang area of China. The strains, designated 8-2(T), W11-1 and 25-7(T), were motile, spore-forming, aerobic rods and contained meso-diaminopimelic acid in their cell walls. Their DNA G+C contents were 37.4, 37.2 and 39.9 mol%, respectively. The main fatty acids in the cellular membranes of these novel strains were C(15) and C(17) methyl-branched. No species with validly published names showed 16S rRNA gene sequence similarity of more than 95 % with respect to these novel isolates; the most closely related species was a halophilic denitrifier, Bacillus halodenitrificans (94.6 %). Polyphasic taxonomic studies revealed that these strains belong to the Bacillaceae and are distantly related to other genera of the family. It is proposed that a new genus, Salinibacillus, should be created, with Salinibacillus aidingensis (type strain, 25-7(T)=AS 1.3565(T)=JCM 12389(T)) as the type species. Another species, Salinibacillus kushneri, is also proposed, with 8-2(T) (=AS 1.3566(T)=JCM 12390(T)) as the type strain.

Bacillaceae↗

Vulcanibacillus modesticaldus gen. nov., sp. nov., a strictly anaerobic, nitrate-reducing bacterium from deep-sea hydrothermal vents.

A novel anaerobic, moderately thermophilic, spore-forming bacterium, designated strain BRT, was isolated from deep-sea hydrothermal core samples collected at the Rainbow vent field on the Mid-Atlantic Ridge (36 degrees 14' N 33 degrees 54' W). The cells were found to be rod-shaped, non-motile, Gram-positive and spore-forming. The organism grew in the temperature range 37-60 degrees C, with an optimum at 55 degrees C, and at pH values in the range 6-8.5, with an optimum around pH 7. NaCl concentrations for growth were in the range 10-40 g l(-1), with an optimum at 20-30 g l(-1). Strain BRT grew chemo-organoheterotrophically with carbohydrates, proteinaceous substrates and organic acids with nitrate as electron acceptor. The novel isolate was not able to ferment. The G+C content of the genomic DNA was 34.5 mol%. Phylogenetic analysis of the 16S rRNA gene sequence placed strain BRT in the Bacillaceae within the class 'Bacilli'. On the basis of the phenotypic and phylogenetic data, this isolate should be described as a member of a novel genus, for which the name Vulcanibacillus gen. nov. is proposed. The type species is Vulcanibacillus modesticaldus sp. nov., with the type strain BRT (=DSM 14931T=JCM 12998T).

Atlantic Ocean↗