Differential diagnosis of Bacillus cereus, Bacillus anthracis, and Bacillus cereus var. mycoides.
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Bacillus↗
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beta-Amylase production by some Bacillus cereus, Bacillus megaterium and Bacillus polymyxa [correction of polymaxa] strains.
The production of extracellular beta-amylase by some Bacillus cereus, Bacillus megaterium and Bacillus polymyxa [corrected] strains was investigated, and the maximal yields of the enzyme were 3.6; 9.3 and 20.4 U/mL of the culture fluid, respectively (U, 1 mumol of maltose equivalent per min at 30 degrees C). Several cultivation media were used for beta-amylase production. Bacillus cereus and some strains of Bacillus megaterium gave good yields of beta-amylase only in medium with the addition of nutrient broth. However, beta-amylase produced during growth in protein rich medium (nutrient broth) was highly unstable, probably due to inactivation by proteolytic enzymes co-existing in the culture fluid. Bacillus polymyxa [corrected] strains can produce good yields of beta-amylase on a semi-synthetic medium consisting of inorganic salts, potato starch and inexpensive soybean extract instead of costly peptone and meat extract. The most potential beta-amylase producer was the strain Bacillus polymyxa [corrected] NCIB 8524. The tested Bacillus megaterium and Bacillus polymyxa [corrected] strains were apparently differentiated by temperature cultivation (30 and 37 degrees C) suitable for beta-amylase amylase yield.
Bacillus↗
Diversity and differential distribution of IS231, IS232 and IS240 among Bacillus cereus, Bacillus thuringiensis and Bacillus mycoides.
Bacillus cereus, Bacillus thuringiensis and Bacillus mycoides are very closely related bacteria, generally considered as subspecies of B. cereus sensu lato. Different transposable elements have been isolated from B. thuringiensis, including IS231, IS232 and IS240 and their variants. The distribution of these three insertion sequences (IS) within the B. cereus group has been investigated in 90 strains of B. thuringiensis (representing 61 serovars), in 30 reference strains of B. cereus and in 33 strains of B. mycoides. Since these IS elements are delimited by well-conserved and specific inverted repeats, the use of primers corresponding to these ends allowed their amplification by PCR. The results showed that IS231 is the most abundant element in the three taxa, whereas IS232 is apparently exclusively associated with B. thuringiensis. Hybridization and Dral RFLP analysis of the PCR products confirmed and extended knowledge of the heterogeneity previously observed among iso-IS231 elements. Moreover, a similar diversity was observed among iso-IS240 elements. This contrasted with the relative homogeneity displayed by iso-IS232 elements. No specific association appeared to exist between any particular iso-element and a specific strain or serotype.
Bacillus↗
Bacillus cereus, Bacillus thuringiensis and Bacillus mycoides differentiation using a PCR-RE technique.
A method was developed to differentiate between Bacillus cereus, Bacillus mycoides and Bacillus thuringiensis using the polymerase chain reaction combined with a restriction endonuclease (PCR-RE) technique. This fast and simple protocol, applied to pure culture strains, was developed using the gyrB DNA sequence, as previously proposed by other authors. Strains from international collections were used to optimize the method which was then applied to the identification of strains isolated from food samples. Amplifications were specific for the B. cereus group. Only Staphylococcus aureus gave the same size PCR product, but it was easily differentiated from strains in the B. cereus group by using restriction analysis, based on digestion with the RsaI, Sau3AI and EcoRI endonucleases. Specific amplifications and good differentiations were obtained using pure strains, suggesting the possibility of using the method described to identify the B. cereus group directly in food samples.
Bacillus↗
A comparative study of Bacillus cereus, Bacillus thuringiensis and Bacillus anthracis extracellular proteomes.
Bacillus cereus, Bacillus thuringiensis and Bacillus anthracis are closely related species that share a similar genetic background but occupy different ecological niches. Virulence plasmids bearing genes coding for toxins, may explain, at least partly, this specialization. We have compared by 2-DE in the early stationary phase of growth the extracellular proteomes of three strains of these species that have lost their virulence plasmids. Proteins expected to be secreted or to belong to the cell wall or to the cytosol were found in the three proteomes. For the cell wall and cytosolic proteins located in the extracellular space, the three proteomes were similar. Cytosolic proteins included enolase, GroEL, PdhB, PdhD, SodA and others. Cell surface proteins were mainly autolysins, proteases, nucleotidases and OppAs. In contrast, the secreted proteins profiles of B. cereus and B. thuringiensis were quite different from that of B. anthracis. B. cereus and B. thuringiensis extracellular proteomes both contained large amounts of secreted degradative enzymes and toxins, including nine proteases, three phospholipases, two haemolysins and several enterotoxins. Most of the genes encoding these enzymes and toxins are controlled by the transcriptional activator PlcR. The extracellular proteome of the pXO1-, pXO2- B. anthracis 9131 strain contained only one secreted protein: the metalloprotease InhA1, also found in the proteomes of the two other strains and possibly involved in antibacterial peptide degradation.
Amino Acid Sequence↗
Comparative analysis of two-component signal transduction systems of Bacillus cereus, Bacillus thuringiensis and Bacillus anthracis.
Members of the Bacillus cereus group are ubiquitously present in the environment and can adapt to a wide range of environmental fluctuations. In bacteria, these adaptive responses are generally mediated by two-component signal transduction systems (TCSs), which consist of a histidine kinase (HK) and its cognate response regulator (RR). With the use of in silico techniques, a complete set of HKs and RRs was recovered from eight completely sequenced B. cereus group genomes. By applying a bidirectional best-hits method combined with gene neighbourhood analysis, a footprint of these proteins was made. Around 40 HK-RR gene pairs were detected in each member of the B. cereus group. In addition, each member contained many HK and RR genes not encoded in pairs ("orphans"). Classification of HKs and RRs based on their enzymic domains together with the analysis of two neighbour-joining trees of these domains revealed putative interaction partners for most of the "orphans". Putative biological functions, including involvement in virulence and host-microbe interactions, were predicted for the B. cereus group HKs and RRs by comparing them with those of B. subtilis and other micro-organisms. Remarkably, B. anthracis appeared to lack specific HKs and RRs and was found to contain many truncated, putatively non-functional, HK and RR genes. It is hypothesized that specialization of B. anthracis as a pathogen could have reduced the range of environmental stimuli to which it is exposed. This may have rendered some of its TCSs obsolete, ultimately resulting in the deletion of some HK and RR genes.
Adaptation, Physiological↗
Heat-stable toxin production by strains of Bacillus cereus, Bacillus firmus, Bacillus megaterium, Bacillus simplex and Bacillus licheniformis.
Strains of Bacillus cereus can produce a heat-stable toxin (cereulide). In this study, 101 Bacillus strains representing 7 Bacillus species were tested for production of heat-stable toxins. Strains of B. megaterium, B. firmus and B. simplex were found to produce novel heat-stable toxins, which showed varying levels of toxicity. B. cereus strains (18 out of 54) were positive for toxin production. Thirteen were of serovar H1, and it was of interest that some were of clinical origin. Two were of serovars 17B and 20, which are not usually implicated in the emetic syndrome. Partial purification of the novel B. megaterium, B. simplex and B. firmus toxins showed they had similar physical characteristics to the B. cereus emetic toxin, cereulide.
Bacillus↗
Cloning and nucleotide sequencing of genes for a second type of small, acid-soluble spore proteins of Bacillus cereus, Bacillus stearothermophilus, and "Thermoactinomyces thalpophilus".
The nucleotide sequences of the single genes coding for the B-type small, acid-soluble spore proteins (SASP) of Bacillus cereus, B. stearothermophilus, and "Thermoactinomyces thalpophilus" were determined, and the amino acid sequences of all B-type SASP were compared. While this type of SASP showed significant sequence conservation around the two spore protease cleavage sites, alignment of these sequences required the introduction of gaps, and even then only 19 of the residues were conserved exactly in all five proteins. However, all five B-type SASP did contain a large (27 to 35-residue), rather well-conserved amino acid sequence repeat, and four of the five proteins had well-conserved regions of 14 to 17 amino acids which appeared three times.
Amino Acid Sequence↗
Growth promotion of red pepper plug seedlings and the production of gibberellins by Bacillus cereus, Bacillus macroides and Bacillus pumilus.
The growth of red pepper plug seedlings was promoted by Bacillus cereus MJ-1, B. macroides CJ-29, and B. pumilus CJ-69 isolated from the rhizosphere. Gibberellins (GAs), a well-known plant growth-promoting hormone, were detected in the culture broth of their rhizobacteria. Among the GAs, the contents of GA1, GA3, GA4, and GA7, physiologically active GAs, were comparatively higher than those of others, suggesting that the growth promoting effect was originated from the GAs. This isthe first report on the production of GA5, GA8, GA34, GA44, and GA53 by bacteria.
Bacillus↗
[The identification of Bacillus cereus, Bacillus lichenformis and Bacillus subtilis strains using the coagglutination reaction].
Polyclonal rabbit antisera against 4 B. cereus strains were selected from a total of 9 B. cereus antisera and pooled. This serum agglutinated all available B. cereus strains (n = 63) at a titre > or = 1:64 when tested by a slide co-agglutination reaction. One hundred and thirty-six Bacillus strains belonging to 17 other species reacted with higher serum concentrations only (titres mainly < or = 1:16). A pooled antiserum comprised of two B. licheniformis antisera and two B. subtilis antisera agglutinated all 43 B. licheniformis strains and all 38 B. subtilis strains at a titre > or = 1:128, but one strain each of the species B. pumilus, B. macerans and B. fastidiosus was also agglutinated at a titre 1:128.
Agglutination Tests↗
[Treatment of iron-deficiency hypochromic anemia with Bacillus cereus & Bacillus coagulans in patients with enterocolitis].
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Anemia↗
MICs of selected antibiotics for Bacillus anthracis, Bacillus cereus, Bacillus thuringiensis, and Bacillus mycoides from a range of clinical and environmental sources as determined by the Etest.
This paper presents Etest determinations of MICs of selected antimicrobial agents for 76 isolates of Bacillus anthracis chosen for their diverse histories and 67, 12, and 4 cultures, respectively, of its close relatives B. cereus, B. thuringiensis, and B. mycoides derived from a range of clinical and environmental sources. NCCLS breakpoints are now available for B. anthracis and ciprofloxacin, penicillin, and tetracycline; based on these breakpoints, the B. anthracis isolates were all fully susceptible to ciprofloxacin and tetracycline, and all except four cultures, three of which had a known history of penicillin resistance and were thought to originate from the same original parent, were susceptible to penicillin. Based on NCCLS interpretive standards for gram-positive and/or aerobic bacteria, all cultures were susceptible to amoxicillin-clavulanic acid and gentamicin and 99% (one with intermediate sensitivity) of cultures were susceptible to vancomycin. No group trends were apparent among the different categories of B. cereus (isolates from food poisoning incidents and nongastrointestinal infections and food and environmental specimens not associated with illness). Differences between B. anthracis and the other species were as expected for amoxicillin and penicillin, with all B. anthracis cultures, apart from the four referred to above, being susceptible versus high proportions of resistant isolates for the other three species. Four of the B. cereus and one of the B. thuringiensis cultures were resistant to tetracycline and a further six B. cereus and one B. thuringiensis cultures fell into the intermediate category. There was a slightly higher resistance to azithromycin among the B. anthracis strains than for the other species. The proportion of B. anthracis strains fully susceptible to erythromycin was also substantially lower than for the other species, although just a single B. cereus strain was fully resistant. The Etest compared favorably with agar dilution in a subsidiary test set up to test the readings, and it compared with other published studies utilizing a variety of test methods.
Anti-Bacterial Agents↗
Cloning and nucleotide sequencing of genes for small, acid-soluble spore proteins of Bacillus cereus, Bacillus stearothermophilus, and "Thermoactinomyces thalpophilus".
As found previously with other Bacillus species, spores of B. stearothermophilus and "Thermoactinomyces thalpophilus" contained significant levels of small, acid-soluble spore proteins (SASP) which were rapidly degraded during spore germination and which reacted with antibodies raised against B. megaterium SASP. Genes coding for a B. stearothermophilus and a "T. thalpophilus" SASP as well as for two B. cereus SASP were cloned, their nucleotide sequences were determined, and the amino acid sequences of the SASP coded for were compared. Strikingly, all of the amino acid residues previously found to be conserved in this group of SASP both within and between two other Bacillus species (B. megaterium and B. subtilis) were also conserved in the SASP coded for by the B. cereus genes as well as those coded for by the genes from the more distantly related organisms B. stearothermophilus and "T. thalpophilus." This finding strongly suggests that there is significant selective pressure to conserve SASP primary sequence and thus that these proteins serve some function other than simply amino acid storage.
Amino Acid Sequence↗
Studies on megacinogeny in Bacillus cereus. II. Bacillus cereus isolates characterized by prophage-controlled production of megacin A (phospholipase A).
Five out of a number of Bacillus cereus strains isolated from soil produced high titre specific bacteriocin (megacin A) in mitomycin C-induced cultures. In the course of cultivation with ethidium bromide, the strains gave off segregants not producing bacteriocin (cin-). The lysate of two wild strains formed plaques on the corresponding cin- bacteria. The two phages (wx23 and wx26) were identical in antigenic structure with phage wx was present in the lysate of B. cereus strain W, and converted cin- derivatives into cultures producing megacin A (phospholipase A). The phages produced plaques at 26 degrees C but not at 37 degrees C. In the lysates of the remaining three strains phages were not detected with biological and morphological methods; these cultures have been assumed to carry defective prophage genome. As the corresponding prophages are responsible for the determination of inducible phospholipase A production, phages named wx seem to form a separate group of B. cereus phages.
Bacillus cereus↗
Bovine abortion caused by Bacillus cereus.
Bacillus cereus was identified as an infrequent abortigenic agent in cattle. Necrotizing placentitis with no or sporadic lesions in fetal tissues was seen. Bacillus cereus was isolated in pure culture from fetal tissue and/or placenta. The recent identification of a bovine abortion caused by B cereus, prompted a retrospective survey of the pathology files. Eight of 947 bovine abortions were attributed to B cereus. Bacillus cereus is often mistaken as a contaminant in bovine abortion because of the failure to identify lesions in fetal tissues compatible with bacterial invasion. A necrotizing toxin may be responsible for the placentitis, with expulsion of the fetus before bacterial colonization.
Abortion, Septic↗
Effect of canavanine from alfalfa seeds on the population biology of bacillus cereus
Bacillus cereus UW85 suppresses diseases of alfalfa seedlings, although alfalfa seed exudate inhibits the growth of UW85 in culture (J. L. Milner, S. J. Raffel, B. J. Lethbridge, and J. Handelsman, Appl. Microbiol. Biotechnol. 43:685-691, 1995). In this study, we determined the chemical basis for and biological role of the inhibitory activity. All of the alfalfa germ plasm tested included seeds that released inhibitory material. We purified the inhibitory material from one alfalfa cultivar and identified it as canavanine, which was present in the cultivar Iroquois seed exudate at a concentration of 2 mg/g of seeds. Multiple lines of evidence suggested that canavanine activity accounted for all of the inhibitory activity. Both canavanine and seed exudate inhibited the growth of UW85 on minimal medium; growth inhibition by either canavanine or seed exudate was prevented by arginine, histidine, or lysine; and canavanine and crude seed exudate had the same spectrum of activity against B. cereus, Bacillus thuringiensis, and Vibrio cholerae. The B. cereus UW85 populations surrounding canavanine-exuding seeds were up to 100-fold smaller than the populations surrounding non-canavanine-exuding seeds, but canavanine did not affect the growth of UW85 on seed surfaces. The spermosphere populations of canavanine-resistant mutants of UW85 were larger than the spermosphere populations of UW85, but the mutants and UW85 were similar in spermoplane colonization. These results indicate that canavanine exuded from alfalfa seeds affects the population biology of B. cereus.
Journal Article↗
A ferric dicitrate uptake system is required for the full virulence of Bacillus cereus.
Bacillus cereus is an opportunistic human pathogen of increasing prevalence. Analysis of the Bacillus cereus genome sequence identified a potential ferric dicitrate uptake system. The three-gene operon was confirmed to be negatively regulated by the ferric uptake repressor (Fur). The Fec operon was genetically silenced using the integration suicide vector pMUTIN4. The mutant strain displayed no growth defect under iron-limited conditions but was unable to grow on ferric citrate as a sole iron source. The virulence of the mutant strain was attenuated in a lepidopteran infection model, highlighting the importance of iron uptake systems to the virulence of B. cereus and the potential of these systems to act as targets for novel antimicrobial agents.
Animals↗
LETHALITY FOR MICE OF VEGETATIVE AND SPORE FORMS OF BACILLUS CEREUS AND BACILLUS CEREUS-LIKE INSECT PATHOGENS INJECTED INTRAPERITONEALLY AND SUBCUTANEOUSLY.
Lamanna, Carl (Army Research Office, Washington, D.C.) and Linda Jones. Lethality for mice of vegetative and spore forms of Bacillus cereus and Bacillus cereus-like insect pathogens injected intraperitoneally and subcutaneously. J. Bacteriol. 85:532-535. 1963.-Lethality for mice of Bacillus thuringiensis var. thuringiensis (two strains), var. sotto (one strain), var. alesti (one strain), and B. entomocidus var. entomocidus (one strain) maintained on artificial laboratory media is dependent upon the route of injection, the number of the bacilli, and their biological state, i.e., vegetative or spore form. Deaths did not occur upon subcutaneous injection of tenfold greater numbers of organisms than the ld(50) dose for the intraperitoneal route. From 10 to 100 times the number of spores were required to kill than vegetative bacilli given intraperitoneally. These observations duplicated those made with strains of B. cereus. For both types of spore forms, from 5 x 10(5) to 5 x 10(7) vegetative organisms were required for one intraperitoneal ld(50). With two strains of highly virulent B. anthracis, ld(50) values were the same for the vegetative and spore states, and for subcutaneous and intraperitoneal injections. Thus, the insect pathogenic sporeforming bacteria tested are more like B. cereus than like B. anthracis in their capacity to infect mice. The similarity to B. cereus extends to the rapidity of death after fatal injection of the insect pathogenic strains. The medical and taxonomic implications of the observations are discussed.
Animals↗