Synbiotic Smallanthus Sonchifolius (Yacon) and Streptococcus Salivarius Inhibit Candida Albicans biofilm formation

Oral biofilms are comprised of extracellular polysaccharides and polymicrobial microorganisms. The objective of this study was to determine the effect of synbiotic Smallanthus sonchifolius (yacon) with Streptococcus salivarius K12 (K12) on polymicrobial biofilm formation of Candida albicans with the hypothesis that polymicrobial biofilm biomass of C. albicans is inhibited by synbiotic S. sonchifolius with K12. Initially, disk diffusion and well diffusion assay were conducted to determine the susceptibility of C. albicans towards S. sonchifolius and K12. Following that, C. albicans was mixed with S. salivarius in nutrient broth (NB) or RPMI-1640 to determine the effect of probiotic on the polymicrobial biofilm. To determine the effect of synbiotic, similar protocol was repeated by adding 800 mg mL-1 of S. sonchifolius aqueous extract extract into the same followed by a 72 h incubation. Finally, biofilm biomass was measured using a crystal violet assay. C. albicans ATCC MYA-4901, ALC2 and ALC3 were found to be susceptible to S. sonchifolius extract and S. salivarius K12. However, the biofilm of all of C. albicans strains ATCC MYA-4901, ALC2 and ALC3 were found to reduce ranged in between 20% to 39.4% when co-cultured with synbiotic compared to prebiotic culture in NB. In conclusion, synbiotic S. sonchifolius with K12 inhibit polymicrobial biofilm. This indicates the potential use of synbiotic in dental application for the prevention C. albicans infection

Synbiotic Musa acuminata skin extract and Streptococcus salivarius K12 inhibit candida species biofilm formation

This study aimed to determine the effect of synbiotic Musa acuminata skin extract (MASE) and Streptococcus salivarius K12 (K12) on Candida species biofilm formation. Liquid chromatography quadrupole time-of-flight (LC-Q-TOF-MS) was conducted to characterize MASE. To determine the effect of synbiotic on Candida biofilm, 200 mL of RPMI-1640 containing Candida, K12, and MASE were pipetted into the same well and incubated at 37�C for 72h. A similar protocol was repeated with K12 or MASE to determine the probiotic and prebiotic effects, respectively. Dimorphism, biofilm biomass, and Candida total cell count (TCC) were determined. A total of 60 compounds were detected in MASE. C. albicans (ALT5) and Candida lusitaniae exhibited the highest reduction in biofilm biomass when co-cultured with prebiotic (77.70 ± 7.67%) and synbi- otic (97.73 ± 0.28%), respectively. All Candida spp. had decreased TCC and hyphae when co-cul- tured with synbiotic. In conclusion, MASE and K12 inhibit Candida biofilm formation

Isolation of Candida species in children and their biofilm-forming ability on nano-composite surfaces

Candida species including Candida albicans, Candida krusei and Candida glabrata are opportunistic microorganisms that inhabit oral cavity. The objective of this study is to determine the effect of dental caries on Candida spp. biofilm-forming ability on nano-composite with the hypothesis that dental caries enhances the colonization of Candida spp. To assess Candida spp. colonisation in the oral cavity of the paediatric patient, samples were obtained from 30 subjects aged five to six years old from Kuantan, Pahang, Malaysia. The samples were collected from buccal mucosa, palate and tooth surfaces using sterile swabs. 10 mL of patient’s saliva suspension was also collected. Following that, the samples were inoculated on CHROMagar and incubated for 24 h at 37 ºC. Candida biofilm of caries isolate C. albicans (HNFC2), and C. albicans ATCC 32354 were developed on three different types of nano-composites. The study showed that no C. albicans was isolated from the caries-free oral cavity while 76% of children with caries possessed Candida spp. 65% of the yeasts were isolated from the tooth surface. Only 35% of the total isolates were obtained from soft tissues, including palatal and buccal mucosa. C. albicans is the most isolated Candida spp. with 82% and 67% of the yeast were obtained from the tooth surface and buccal mucosa, respectively. Besides, HNFC2 significantly colonised the nano-composites more than the ATCC (P < 0.05). In the comparison of the three types of nano-composites, nano-hybrid-based containing pre-polymerised filler (cB) exhibited the least C. albicans HNFC2 cells colonisation with 7.7 x 10³ cells mL-1. In contrast, the nano-composite that contained bulk-filled nanohybrid (cC) was the most colonised with 14.3 x 10³ cells mL-1. In conclusion, dental caries enhances the colonization of Candida spp. in children's oral cavity, and that caries isolate form more biofilm on nano-composites compared to the lab strain C. albicans

The effect of synbiotic Streptococcus salivarius K12 and Yacon (Smallanthus sonchifoliuson) on Candida albicans biofilm formation

Candida albicans is an opportunistic fungus that is known for its ability to form biofilms. Streptococcus salivarius K12 is an oral probiotic while yacon is a source of prebiotic. The objective of this study is to investigate the effect of S. salivarius K12 and yacon aqueous extract (synbiotic) on C. albicans with the hypothesis that S. salivarius K12 and yacon inhibit C. albicans biofilm formation. To develop mono-species biofilm, C. albicans (ATCC MYA-4901 and cancer isolates, ALC2 and ALC3 strains) and S. salivarius K12 were standardised to 105 cells and 106 cells, respectively and grown in 96-well plate in nutrient broth (NB) or RPMI at 37 °C for 72 h. Polymicrobial biofilms were developed by inoculating both microorganisms in the same well with similar cell number as in mono-species. To determine the effect of the synbiotic, a similar protocol was repeated by mixing with 800 mg mL-1 of yacon extract and incubated at 37 °C for 72 h. The medium was replenished at every 24 h, aseptically. Finally, the biofilms were assessed using the crystal violet assay, and the optical density was measured at OD620nm. The combination of both prebiotic and probiotic has effectively reduced all the C. albicans strain (MYA-4901, ALC2 and ALC3) in both NB and RPMI. All C. albicans strain when grown in polymicrobial with S. salivarius K12 in NB that is predominated by yeast-form C. albicans, exhibited decreased biofilms by 51.34±11.6, 8.20±43.9 and 11.3±82.7%, respectively when compared to the expected biofilms. Meanwhile in RPMI, which C. albicans strain ATCC MYA-4901, ALC2 and ALC3 were predominated by hyphal-form showed decreased biofilms by 43.3±12.1%, 39.4±15.7% and 25.7±56.8%, respectively when compared to the expected biofilms. S. salivarius K12 and yacon extract synbiotic inhibits biofilm formation of C. albicans yeast and hyphal forms thus supported the hypothesis of the present study

Suppression of non-albicans candida species (NAC) biofilm formation by probiotic Streptococcus salivarius

Introduction: Candida spp. are most common opportunistic pathogenic yeast that inhabit human oral cavity, epider­mis, gastrointestinal tract, and vagina leading to candidiasis. The transition of this yeast from commensal to potent pathogen is facilitated by numbers of virulence factors including biofilm formation. While most reports on candidi­asis are associated with formation Candida albicans biofilms, however, non-albicans Candida species prevalence is of growing concern. Recently, the use of probiotics as antifungal and antibiofilm has gained an increasing attention. As such, we aim to evaluate the inhibitory effect of monomicrobial and polymicrobial of Streptococcus Salivarius on six strains of NAC namely Candida dubliniensis, Candida glabrata, Candida krusei, Candida lusitanaei, Candida parapsilosis and Candida tropicalis. Methods: Antifungal activity of S. salivarius on NAC species was performed using well diffusion method on Mueller Hinton Agar (MHA) and the diameter of inhibition zone were assessed. For formation of monomicrobial biofilm, standardized cell suspensions of NAC species and probiotic Streptococcus salivarius were grown in RPMI or nutrient broth media at 37°C for 72 h. Meanwhile to study polymicrobial biofilm -of both NAC and 5. salivarius, similar protocol was employed by inoculating both microorganisms with a similar cell density as in monomicrobial. Finally, biofilrm formation was assessed through quantification of total biomass by crystal violet (CV) assay and the absorbance of adherent biofilm was measured in triplicate at 620nm. Results: Antifungal susceptibility testing of S. salivarius on all six NAC species discerned no zone of inhibition. Furthermore, our results showed variability of monomicrobial and polymicrobial biofilm biomass between NAC species and growth medium. All six polymicrobial NB-grown and RPMl-grown exhibited decreased of the biofilm formation. C. parapsilosis co-cultured with 5. salivarius in NB medium had shown lowest biofilm bio­mass by 75.51 + _1 .34% while in RPMI medium, C. lusitanaei demonstrated with most reduced biofilm biomass by 67.03+ _5. 19. Conclusion: Our study elucidated the antagonistic relationship between Streptococcus salivarius and non-albicans Candida by supressing the growth of polymicrobial biofilm and pseudohyphae/hyphae of NAC species

The effect of synbiotic Streptococcus salivarius K12 and Musa acuminata on Candida albicans biofilm formation

Introduction: Candida a/bicans is an opportunistic fungus that is associated with oral carcinogenesis. In addition, biofilm formation has been one of the important virulence factors of the yeast. Streptococcus salivarius Kl 2 is an oral probiotic while Musa acuminata is a well-known prebiotic. The objective of this study is to investigate the effect of 5. salivarius IK1 2 and M. acuminata skin aqueous extract (synbiotic) on C. albicans with the hypothesis that S. salivarius Kl 2 and M. acuminata inhibit C. a/bicansbiofilm formation. Methods: To develop mono-species biofilnn, C. a/bicans (A TCC MYA-4901 and cancer isolates, ALC2 and ALC3 strains) and 5. sa/ivarius Kl 2 were standardised to 1 OS cells and 106 cells, respectively and grown in 96-well plate iin nutrient broth (NB) or RPMI at 37 cc for 72 h. Polymicro­bial biofilms were developed by inoculating both microorganisms in the same well with similar cell number as in mono-species. To determine the effect of synbiotic, similar protocol was repeated by mixing with 800 mg ml-1 of M. acuminata skin extract and incubated at 37 cc for 72 h. The medium was replenished at every 24 h, aseptically. Finally, the biofilms were assessed using crystal violet assay and the optical density was measured at 00620nm. Results: C albicans strain MYA-4901 and ALC3, when grown in polymicrobial with 5. salivarius Kl 2 in NB that is predominated by yeast-form C. albicans, exhibited decreased biofilms by 71.40± 11.7% and 49.40±3.9%, respec­tively when compared to the expected biofilms. Meanwhile in RPMI, which C. a/bicansstrain ATCC MYA-4901, ALC2 and ALC3 were predominated by hyphal-form showed decreased biofilms by 72.0±26.7%, 53.4± 14.4% and 65.7±6.7%, respectively when compared to the expected biofilms. Conclusion: 5. sa/ivarius Kl 2 and M. acuminata skin extract syn biotic inhibit biofilm formation of C. a/bicans yeast and hyphal forms thus supported tlhe hypothesis of the present study

Characterization of Cervus timorensis velvet antler and its effect on biofilm formation of Candida species

Oral biofilms comprise extracellular polysaccharides and polymicrobial microorganisms. The objectives of the study were to characterize the deer velvet antler (DVA) compounds and their effect on Candida species biofilm formation with the hypothesis that DVA inhibits the biofilm of Candida spp. Liquid Chromatography-Quadrupole Time of Flight-Mass Spectrometry (LC-QTOF-MS) was conducted to characterize the DVA compounds. To study the effect of DVA on biofilm, Candida albicans ATCC MYA-4901 (ALT5), AIDS isolate (ALC2), oral cancer isolate (ALC3), C. dubliniensis ATCCMYA-2975,C.glabrataATCC90030,C.krusei14 243,C.lusitaniaeATCC34449,C.parapsilosisATCC22019,andC.tropicalisATCC13803 were inoculated with DVA in separate wells of a 96-well plate containing RPMI-1640 followed by 72 h incubation. A total of 45 compounds were detected in the DVA extract. C. lusitaniae exhibited a higher percentage of biofilm biomass reduction when treated with DVA extract (66.10% ± 5.33), followed by ALC3 (44.12% ± 6.24). However, C. glabrata, C. krusei, and C. parapsilosis showed no reduction in biofilm biomass after being treated with DVA extract. Most Candida strains also exhibited decreased total cell count when treated with DVA extract, except for ALC3 and C. krusei. ALT5 had the lowest total cell count (0.17 × 105 cells/ml) when cultured with DVA extract. In conclusion, DVA extract inhibits Candida spp. biofilm formation except for C. glabrata, C. krusei, and C. parapsilosis