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Login Register Login using. You can login by using one of your existing accounts. We will be provided with an authorization token please note: passwords are not shared with us and will sync your accounts for you. The organisation of the P. The percentage identity of the product of each putative gene to their P. ClustalX alignments of putative B. Diamonds indicate a potential Fe-S binding site at 4 cysteine residues in HcnA. We looked for cyanide production by B.
The pH of the culture media was measured and was always around pH 8, which is the same as that seen for P. This suggests that cyanide was not being lost from the culture as hydrogen cyanide gas due to acidification of the culture medium. Glycine is the precursor for hydrogen cyanide synthesis in P.
However, no cyanide was detected when B. Oxygen is a regulator of cyanide synthesis in P. However, despite varying the O 2 -transfer coefficient k L a values from We conclude that planktonic cultures of B. Cyanide production by P. Under these colonial growth conditions cyanide production was detected from both B. The concentrations of cyanide trapped were 8. However, when normalised to the CFU count of the culture cyanide production from both B. This demonstrated that B.
Concentration of cyanide given off by plate grown culture and trapped in NaOH.
Professor Brajesh Singh
Concentration of trapped cyanide normalised to the CFU count for the plate. Values are the averages of 3 independent replicates with SE error bars. We next screened other members of the Bcc for cyanide production. A total of 34 strains with at least one representative from each species genomovar were assayed using the LB agar plate method Table 1.
Every strain tested was cyanogenic except for two B.
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In this assay a P. The limited number of replicates for some strains meant that mean cyanide production could only be compared between B. The mean normalised cyanide concentration for the B. All strains were also assayed for cyanide production in liquid culture, but like K and J none of them tested positive. Cyanide production by members of the Bcc grown on solid media. Concentration of cyanide given off by plate grown cultures of Bcc strains. Concentration of cyanide trapped in NaOH.
Cyanide concentrations from A normalised to the CFU count for the plate. The numbers under the axis correspond to stain names for key to strain names see Table 1.
In summary, all nine species in the Bcc are capable of producing cyanide when grown on solid media. The level of cyanide production in the Bcc appears to be strain rather than species specific. The finding that cyanide was not detected in liquid cultures of B.
Bcc can grow as a biofilm [ 10 , 27 ] and we were interested to test whether B. We grew Bcc strains as biofilms on 6 mm diameter glass beads immersed in LB broth in a Petri dish, which provided a large surface area for growth. Biofilm growth was detected visually and by crystal violet staining [ 10 , 27 ]. Cyanide production was assessed by trapping cyanide given off from the biofilm culture in 4 M NaOH.
Control cultures comprised Petri dishes containing bacterial culture without glass beads. Figure 4A shows the data obtained for the P. Cyanide was detected in the presence and absence of glass beads, consistent with P. Cyanide production from glass bead "biofilm" model. Concentration of cyanide given off by glass bead biofilm cultures over 6 hours and trapped in 1 ml 4 M NaOH.
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Solid bars: with glass beads, diagonal striped bars: without glass beads. Measurements are averages of 3 independent replicates with SE error bars. For the B. Cyanide was not detected until day 2 or day 3 for K and J respectively. This is consistent with cyanide production resulting from biofilm growth, which takes time to establish on the glass bead surface.
Providing B. This, together with the findings that Bcc strains produce detectable cyanide when grown on solid media but not in liquid culture, suggests that cyanide production in B. In this paper we have demonstrated that B. This is the first time that any member of the Bcc has been reported to produce cyanide, although there are published reports of unsuccessful attempts to detect cyanide production by B.
We can only surmise that strain or growth condition differences prevented the detection of cyanide production in these previous studies. Like P. Therefore, this finding is potentially significant in terms of CF lung infection, especially in light of our recent finding that cyanide accumulates in the lungs of P.