Influence of the Provox Flush, blowing and imitated coughing on voice prosthetic biofilms in vitro.

OBJECTIVE: This study investigates the effect of regular airflow, as an isolated single factor, through Groningen and Provox2 voice prostheses on biofilm formation. MATERIAL AND METHODS: Groningen and Provox2 voice prostheses were placed in a modified Robbins device and inoculated with the total microflora from an explanted Groningen voice prosthesis. After 3 days, prostheses were either flushed 3 times per day with the Provox flush, treated with an airflow using an increasing order of air pressure (10, 15 and 20 cmH2O) or vigorously perfused by means of imitated coughing (air pressure 20 cmH2O). As a control, prostheses were left undisturbed to promote biofilm growth. Following flushing, blowing or coughing, each artificial throat was perfused with 200 ml of phosphate-buffered saline. This procedure was repeated three times a day for 9 days. At the end of each day, the artificial throats were filled with growth medium for 30 min and left empty during the night after draining. After 12 days the microflora on each voice prosthesis was quantified by plating on blood agar for bacteria and on de Man, Rogosa and Sharpe agar for yeasts. RESULTS: The use of the Provox flush reduced bacterial prevalence on Groningen and Provox2 voice prostheses to 71% and 45% of the control values, respectively, without affecting the number of yeasts. Increasing airflows and imitated coughing yielded reductions of 45-70% in bacterial and yeast prevalence on Provox2 voice prostheses. On the Groningen voice prostheses the effects of increasing airflows and imitated coughing were less pronounced: reductions in bacterial and yeast prevalence of 56-87% were observed. CONCLUSION: This study shows that use of the Provox flush has a cleansing effect, especially on Provox2 voice prostheses, and furthermore suggests that daily airflow through voice prostheses as part of a daily maintenance scheme reduces biofilm formation and can be expected to prolong the life of these devices.

Biofilm formation on voice prostheses: in vitro influence of probiotics.

In order to determine the influence of probiotic bacteria on biofilm formation on Groningen and Provox 2 voice prostheses in an artificial throat, we grew biofilms on both types of voice prostheses and exposed them 3 times daily to a probiotic bacterial suspension. As a control, we perfused an artificial throat with phosphate-buffered saline solution. Perfusion with Lactococcus lactis 53 suspension reduced the percentage numbers of bacteria and yeasts, respectively, on the Groningen prostheses to 17% and 22% and on the Provox 2 prostheses to 19% and 45%, compared to the number of colony-forming units on the control prosthesis, which was set at 100%. A suspension of Streptococcus thermophilus B reduced the percentage numbers of bacteria and yeasts, respectively, on the Groningen prostheses to 53% and 33% and on the Provox 2 prostheses to 14% and 0%, as compared to the control prosthesis. All other probiotic strains tested caused some reduction in the percentages of bacteria or yeasts, but strong differences between the types of prostheses were observed. In conclusion, L. lactis 53 and S. thermophilus B strongly reduce the occurrence of yeasts and bacteria in voice prosthetic biofilms.

Air-flow resistances of silicone rubber voice prostheses after formation of bacterial and fungal biofilms.

Laryngectomized patients use silicone rubber voice prostheses to rehabilitate their voice. However, biofilm formation limits the lifetime of voice prostheses by causing leakage or an increased air-flow resistance and the prosthesis has to be replaced. To determine which bacterial or yeast strains, isolated from explanted voice prostheses, contribute most to increases in air-flow resistance of silicone rubber voice prostheses, biofilms consisting of either a bacterial or a yeast strain were grown on voice prostheses in the artificial throat model. The effects of these biofilms on air-flow resistances were determined by calculating the difference in air-flow resistance of the individual voice prosthesis as covered with a 7-day-old biofilm with the situation prior to biofilm formation. Conspicuously, voice prosthetic biofilms formed by the bacterial strains Staphylococcus aureus GB 2/1 and Rothia dentocariosa GBJ 41/25B and their excreted organic matter showed larger increases in air-flow resistance (more then 30 cm H(2)O.s/L) than biofilms formed by Candida species. This is contrary to the literature, where there seems to be agreement that Candida species are mainly responsible for clinical failure of silicone rubber voice prostheses.

Effect of probiotic bacteria on prevalence of yeasts in oropharyngeal biofilms on silicone rubber voice prostheses in vitro.

The proliferation of yeasts in the mixed bacterial and fungal biofilms colonising silicone rubber voice prostheses in laryngectomised patients is the main cause of malfunctioning of the valve mechanism on the oesophageal side of the prostheses. Indwelling voice prostheses usually have to be replaced every 3-4 months. The consumption of probiotic bacteria is largely motivated by health claims related to the urogenital and lower digestive tract, but not to the upper digestive tract. The present study examined the influence of probiotic bacteria on the prevalence of yeasts in oropharyngeal biofilms on silicone rubber voice prostheses, as formed in a modified Robbins device. Exposure of oropharyngeal biofilms on voice prostheses to suspensions of Bifidobacterium infantis 420 or Enterococcus faecium 603 did not significantly reduce the number of yeasts in the biofilm. However, suspensions of Lactobacillus fermentum B54, L. rhamnosus 744 or L. lactis cremoris SK11 led to a reduction in the number of yeasts harvested from the voice prostheses. Suspensions of L. casei Shirota and Streptococcus thermophilus B significantly reduced the number of yeasts in the biofilm to 39% and 33%, respectively. The reduction brought about in yeast prevalence in the mixed biofilm was greatest by exposure to a suspension of L. lactis 53, with yeast prevalence only 4% of the control. In conclusion, the study demonstrated that the prevalence of yeasts in oropharyngeal biofilms on silicone rubber voice prostheses might be controlled by consumption of probiotic bacteria.

Antimicrobial activity of synthetic salivary peptides against voice prosthetic microorganisms.

OBJECTIVES: To investigate whether synthetic salivary antimicrobial peptides have an inhibitory effect on the growth of bacteria and yeasts isolated from used silicone rubber voice prostheses. METHODS: The antimicrobial activities of six synthetic salivary peptides (histatin 5, dhvarl, dhvar4, dhvar5, lactoferrin b 1730 [LFb 17-30], and cystatin S1-15) at concentrations of 2 and 4 mg/mL were determined against different oropharyngeal yeast (four) and bacterial (eight) strains and against a "total microflora" isolated from explanted voice prostheses using agar diffusion tests. The spectrum of susceptible microorganisms was determined qualitatively. RESULTS: Histatin 5 and cystatin S1-15 did not show any antimicrobial activity against the microorganisms involved in this study. Dhvar1 was active against some of the oropharyngeal microorganisms tested, including the yeast strains, but not against Rothia dentocariosa, Staphylococcus aureus, Escherichia coli, and the total microflora Dhvar4 was active against all microorganisms tested, including the total microflora. Dhvar5 lacked activity against E coli and the total microflora LFb 1730 did not inhibit the growth of any of the yeast strains involved and showed only minor activity against some of the bacterial strains. LFb 1730 slightly inhibited the growth of the total microflora from an explanted prosthesis. CONCLUSIONS: The synthetic salivary peptide dhvar4 has a broad antimicrobial activity against all microorganisms that are commonly isolated from explanted voice prostheses, including yeasts. Therewith, it may represent a useful drug, as an alternative for antibiotics and antimycotics employed in various ways to prolong the lifetime of voice prostheses in laryngectomees.