Numerical Modeling of Mass Transfer in the Interaction between River Biofilm and a Turbulent Boundary Layer

In this article dedicated to the modeling of vertical mass transfers between the biofilm and the bulk flow, we have, in the first instance, presented the methodology used, followed by the presentation of various results obtained through analyses conducted on velocity fields, different fluxes, and overall transfer coefficients. Due to numerical constraints (resolution of relevant spatial scales), we have restricted the analysis to low Schmidt numbers (Sc=0.1, Sc=1, and Sc=10) and a single roughness Reynolds number (Re*=150). The analysis of instantaneous concentration fields from various simulations revealed logarithmic concentration profiles above the canopy. In this zone, the concentration is relatively homogeneous for longer times. The analysis of results also showed that the contribution of molecular diffusion to the total flux depends on the Schmidt number. This contribution is negligible for Schmidt numbers Sc≥0.1, but nearly balances the turbulent flux for Sc=0.1. In the canopy, the local Sherwood number, given by the ratio of the total flux (within or above the canopy) to the molecular diffusion flux at the wall, also depends on the Schmidt number and varies significantly between the canopy and the region above. The exchange velocity, a purely hydrodynamic parameter, is independent of the Schmidt number and is on the order of 10% of in the present case. This study also reveals that nutrient absorption by organisms near the wall depends on the Schmidt number. Such absorption is facilitated by lower Schmidt numbers.

Biofilm formation under high temperature causes the commensal bacteria Bacillus cereus WPySW2 to shift from friend to foe in Neoporphyra haitanensis in vitro model

Although biofilm formation may promote growth,biofilms are not always beneficial to their hosts.The biofilm formation characteristics of Bacillus cereus WPySW2 and its changes at different temperatures were studied.Results show that B.cereus WPySW2 promoted the growth of Neoporphyra haitanensis(an economically cultivated seaweed)at 20℃ but accelerated algal rot at 28℃.Thicker B.cereus WPySW2 biofilms covered the surface of N.haitanensis thalli at 28℃,which hindered material exchange between the algae and surrounding environment,inhibited algal photosynthesis and respiration,and accelerated algal decay.Compared with planktonic bacteria,mature biofilm cells had lower energy consumption and metabolic levels.The biofilm metabolic characteristics of B.cereus WPySW2 changed significantly with temperature.High temperature accelerated biofilm maturation,which made it thicker and more stable,allowing the bacteria to easily adapt to environmental changes and obtain greater benefits from their host.High temperature did not affect the production or increased the abundance of toxic metabolites,indicating that the negative effects of B.cereus WPySW2 on algae were not caused by toxins.This study shows that increased temperature can transform a harmless bacterium into a detrimental one,demonstrating that temperature may change the ecological function of phycospheric bacteria by affecting their morphology and metabolism.

Antimicrobial Activities of Extracts of Macrosphyra longistyla against Gram-Positive Oral Biofilm-Formers from School Children in Southwestern Nigeria and Toxicity Studies Using Brine Shrimps

The world will benefit from more effective antimicrobial agents against oral conditions arising from the actions of biofilm forming bacteria. Also, information is lacking on the oral biofilm-forming bacterial diversity in Southwestern Nigeria. In this study, we isolate and characterize oral biofilm producing bacteria in the oral cavities of schoolchildren in Southwestern Nigeria. We also investigate the antimicrobial properties of Macrosphyra longistyla extracts against the biofilm-formers and the toxicity of potent extracts. Samples were obtained from 109 schoolchildren aged 4 - 14 years from Lagos, Oyo and Osun States. Agar well diffusion technique was used in the antimicrobial susceptibility testing. Toxicity testing was done using brine shrimps (Artemia salina). Biofilm-formers in this study are Klebsiella sp., Streptococcus sp., Staphylococcus sp., and Micrococcus sp. Ethanol leaf extracts had the highest activity against all biofilm-producing bacteria. Ethanol stem bark extract, which elicited activity against Klebsiella only, was found to be less toxic than the ethanol leaf extract. Staphylococcus showed >10 mm susceptibility to the ethanol and aqueous extracts of Macrosphyra longistyla. Streptococcus and Micrococcus were susceptible to the antimicrobial actions of the ethanolic leaf extracts. Although the ethanol extracts of the leaves had lower minimum inhibitory concentrations than the ethanol extracts of the stem bark, toxicity studies showed ethanol extracts of the stem-bark to be more toxic than the ethanol extracts of the leaves. In conclusion, ethanolic extracts of Macrosphyra longistyla show potential as sources of antimicrobials against gram-positive, oral biofilm-forming bacteria.

Bacterial Exofactors Modulate Biofilm Growth and Resistivity to Antimicrobial Drugs

Some bacteria have the ability to co-exist, proliferate and survive in a multicellular community, biofilm. Each participating bacteria can form its colonies and encases itself by a self-produced insoluble extracellular matrix substance (EPS). Microcolonies within biofilm are held together by interactions and bonding of the substances present in the EPS with their separation from the water channels. Similar to insoluble EPS, bacterial microcolonies release soluble exofactors that have direct impacts on the survivability, growth and antibacterial resistivity of other microcolonies made of single- or multi-species bacteria in the same biofilm. How the exofactors of microcolonies of one-type bacteria impact on microcolonies of other-type bacteria is still unclear. We studied about the role of exofactors released from Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa, which are common biofilm-forming pathogenic bacteria. Exofactors facilitate to transform the microenvironment where bacteria can acquire alternative lifestyle with a long survival period and resistivity to certain antimicrobial drugs.

Staphylococcus aureus and biofilms:transmission, threats, and promising strategies in animal husbandry

Staphylococcus aureus(S. aureus) is a common pathogenic bacterium in animal husbandry that can cause diseases such as mastitis, skin infections, arthritis, and other ailments. The formation of biofilms threatens and exacerbates S. aureus infection by allowing the bacteria to adhere to pathological areas and livestock product surfaces, thus triggering animal health crises and safety issues with livestock products. To solve this problem, in this review, we provide a brief overview of the harm caused by S. aureus and its biofilms on livestock and animal byproducts(meat and dairy products). We also describe the ways in which S. aureus spreads in animals and the threats it poses to the livestock industry. The processes and molecular mechanisms involved in biofilm formation are then explained. Finally, we discuss strategies for the removal and eradication of S. aureus and biofilms in animal husbandry, including the use of antimicrobial peptides, plant extracts, nanoparticles, phages, and antibodies. These strategies to reduce the spread of S. aureus in animal husbandry help maintain livestock health and improve productivity to ensure the ecologically sustainable development of animal husbandry and the safety of livestock products.

Evolution of Biofilm and Its Effect on Microstructure of Mortar Surfaces in Simulated Seawater

To explore the role of biofilm formation on the corrosion of marine concrete structures, we investigated the attachment of biofilm on mortar surfaces in simulated seawater and the influence of biofilm on the microstructure of mortar surfaces. The results show that the evolution of biofilm on mortar surfaces in simulated seawater is closely related to the corrosion suffered by the mortar, and the process of biofilm attachment and shedding is continuous and cyclical. It is found that the specimens in the absence of biofilm attachment are more severely eroded internally by the corrosive medium in simulated seawater than those in the presence of biofilm attachment. For the specimens without biofilm attachment, after 60 days, gypsum forms,and after 120 days, the number of pores in the mortar is reduced. In contrast, for the specimens in the presence of biofilm attachment, gypsum could only be detected after 90 days, and fewer pores are filled. Therefore, the formation of biofilm could delay the invasion of the corrosive medium into the interior of mortar during the evolution of biofilm on mortar surfaces, mitigating the corrosion of mortars in seawater.

Knowledge and the Attitude on the Use of Mouthwash among Two Selected Senior High Schools in Kumasi

BACKGROUND: The route of most systemic diseases begins in the oral cavity. Oral health knowledge of mouthwashes and their uses is indispensable for the general population and especially adolescents. The use of mouthwash by adolescents can be a beneficial adjunct to their oral hygiene routine, providing additional protection against dental diseases and promoting fresher breath. Overuse or misuse of mouthwash, particularly those containing alcohol or other potentially irritating ingredients, may lead to adverse effects such as oral mucosal irritation, dry mouth, or alteration of the oral microbiome. OBJECTIVES: To determine the knowledge, attitude, and use of mouthwash among senior high school students in Kumasi. METHODOLOGY: 120 students responded to a standard questionnaire by a convenient sample technique. The Statistical Package for Social Sciences (SPSS) 22.0 and MS Excel were used for data management and analysis. The results of the study were presented using tables, bar charts, and pie chart. RESULTS: The ages of the respondents ranged from 14 to 20 years. Out of the 120 participants, 71 students that represent 59.2% of the total, used mouthwash. Majority of the participants (63%) utilized a mouthwash after brushing their teeth. 49% of the participants reported using mouthwash to address halitosis, 37% used it to combat periodontal disease, and 10% used it for relief from a sore throat. CONCLUSION: In general, most of the participants who use mouthwash had excellent knowledge and a positive attitude toward the use of mouthwash.

Structural insights on anti-biofilm mechanism of heated slightly acidic electrolyzed water technology against multi-resistant Staphylococcus aureus biofilm on food contact surface

Slightly acidic electrolyzed water(SAEW)has proven to be an efficient and novel sanitizer in food and agriculture field.This study assessed the efficacy of SAEW(30 mg/L)at 40℃on the inactivation of foodbome pathogens and detachment of multi-resistant Staphylococcus aureus(MRSA)biofilm.Furthermore.the underlying mechanism of MRS A biofilm under heated SAEW at 40℃treatment on metabolic profiles was investigated.The results showed that the heated SAEW at 40℃significantly effectively against foodbome pathogens of 1.96-7.56(lg(CFU/g))reduction in pork,chicken,spinach,and lettuce.The heated SAEW at 40℃treatment significantly reduced MRS A biofilm cells by 2.41(lg(CFU/cm^(2))).The synergistic effect of SAEW treatment showed intense anti-biofilm activity in decreasing cell density and impairing biofilm cell membranes.Global metabolic response of MRSA biofilms,treated by SAEW at 40℃,revealed the alterations of intracellular metabolites,including amino acids,organic acid,fatty acid,and lipid.Moreover,signaling pathways involved in amino acid metabolism,energy metabolism,nucleotide synthesis,carbohydrate metabolites,and lipid biosynthesis were functionally disrupted by the SAEW at 40℃treatment.As per our knowledge,this is the first research to uncover the potential mechanism of heated SAEW treatment against MRSA biofilm on food contact surface.

The clinical impact of bacterial biofilms

Bacteria survive in nature by forming biofilms on surfaces and probably most, if not all, bacteria （and fungi） are capable of forming biofilms. A biofilm is a structured consortium of bacteria embedded in a self-produced polymer matrix consisting of polysaccharide, protein and extracellular DNA. Bacterial biofilms are resistant to antibiotics, disinfectant chemicals and to phagocytosis and other components of the innate and adaptive inflammatory defense system of the body. It is known, for example, that persistence of staphylococcal infections related to foreign bodies is due to biofilm formation. Likewise, chronic Pseudomonas aeruginosa lung infections in cystic fibrosis patients are caused by biofilm growing mucoid strains. Gradients of nutrients and oxygen exist from the top to the bottom of biofilms and the bacterial cells located in nutrient poor areas have decreased metabolic activity and increased doubling times. These more or less dormant cells are therefore responsible for some of the tolerance to antibiotics. Biofilm growth is associated with an increased level of mutations. Bacteria in biofilms communicate by means of molecules, which activates certain genes responsible for production of virulence factors and, to some extent, biofilm structure. This phenomenon is called quorum sensing and depends upon the concentration of the quorum sensing molecules in a certain niche, which depends on the number of the bacteria. Biofilms can be prevented by antibiotic prophylaxis or early aggressive antibiotic therapy and they can be treated by chronic suppressive antibiotic therapy. Promising strategies may include the use of compounds which can dissolve the biofilm matrix and quorum sensing inhibitors, which increases biofilm susceptibility to antibiotics and phagocytosis.

A novel integrated step-feed biofilm process for the treatment of decentralized domestic wastewater in rural areas of China

For wastewater treatment in rural areas, a novel three-stage step-feed wastewater treatment system, combined with a drop-aeration biofilm process, was tested in the laboratory to investigate its performance in removing suspended solids （SS）, chemical oxygen demand （COD）, NH4＋-N, total nitrogen （TN）, and total phosphorus （TP）. The removal rates of SS, COD and NH4＋-N were 90%, 80%, and 90% in efluent concentrations less than 10 mg/L, 50 mg/L and 8 mg/L, respectively. The TP removal rate was less satisfactory. The C/N ratio in the raw wastewater was often less than 3.5, and the removal efficiency of TN was therefore limited. A carbon-release batch experiment was carried out to measure the feasibility of enhancing denitrification at low influent C/N ratios. The result showed that the C/N could be over 9.0 in the supernatant. Polymerase chain reaction denaturing gradient gel electrophoresis technology was used to reveal the changes in the bacterial community during different stages of the integrated step-feed biofilm process. The results showed that banding patterns and the distribution of dominant bands for the same experimental period in different aerobic zones were similar. Phylogenetic analysis indicated that lanes 10, 11 and 12, which presented three aerobic zones at the same operation period, had the closest phylogenetic relationship among the lanes.

The role of bacterial biofilm in persistent infections and control strategies

Bacterial biofilms can be viewed as a specific type of persistent bacterial infection. After initial invasion, microbes can attach to living and non-living surfaces, such as prosthetics and indwelling medical devices, and form a biofilm composed of extracellular polysaccharides, proteins, and other components. In hosts, bioffim formation may trigger drug resistance and inflammation, resulting in persistent infections. The clinical aspects of biofilm formation and leading strategies for hiofilm inhihitors will be discussed in this mini-review.

Effect of biofilm formation by clinical isolates of Helicobacter pylori on the efflux-mediated resistance to commonly used antibiotics

AIM To evaluate the role of biofilm formation on the resistance of Helicobacter pylori(H. pylori) to commonly prescribed antibiotics, the expression rates of resistance genes in biofilm-forming and planktonic cells were compared.METHODS A collection of 33 H. pylori isolates from children and adult patients with chronic infection were taken for the present study. The isolates were screened for biofilm formation ability, as well as for polymerase chain reaction(PCR) reaction with HP1165 and hp1165 efflux pump genes. Susceptibilities of the selected strains to antibiotic and differences between susceptibilities of planktonic and biofilm-forming cell populations were determined. Quantitative real-time PCR(qP CR) analysis was performed using 16 S r RNA gene as a H. pylori-specific primer, and two efflux pumps-specific primers, hp1165 and hefA.RESULTS The strains were resistant to amoxicillin, metronidazole, and erythromycin, except for one strain, but they were all susceptible to tetracycline. Minimum bactericidalconcentrations of antibiotics in the biofilm-forming cells were significantly higher than those of planktonic cells. qP CR demonstrated that the expression of efflux pump genes was significantly higher in the biofilm-forming cells as compared to the planktonic ones.CONCLUSION The present work demonstrated an association between H. pylori biofilm formation and decreased susceptibility to all the antibiotics tested. This decreased susceptibility to antibiotics was associated with enhanced functional activity of two efflux pumps: hp1165 and hefA.

In vitro biofilm formation by uropathogenic Escherichia coli and their antimicrobial susceptibility pattern

Objective:To detect in vitro biofilm formation of uropathogenic Escherichia coli(E.coli)(UPEC) strains isolated from urine specimens and also to determine their antimicrobial susceptibility pattern using 13 commonly used antibiotics.Methods:The present study comprised of 166 urine specimens collected from tertiary care hospitals in and around Coimbatore.South India. All the specimens were subjected to gram staining,bacterial culture and the E.coli strains were screened for biofilm formation using Tube Method(TM),Congo Red Agar(CRA) and Tissue Culture Plate method(TCP) respectively.Subsequently,the antimicrobial susceptibility test was performed by Kirby Bauer-disk diffusion method for the biofilm and non-biofilm producing E. coli strains.Results:Of the 100(60.2%) E.coli strains,72 strains displayed a biofilm positive phenotype under the optimized conditions in the Tube Method and the strains were classified as highly positive(17,23.6%),moderate positive(19.26.3%) and weakly positive(36.50.0%). similarly under the optimized conditions on Congo Red agar medium,biofilm positive phenotype strains were classified as highly positive(23,23%).moderate positive(37.37%) and weakly positive(40,40%).While in TCP method,the biofilm positive phenotype strains were also classified as highly positive(6.6%),moderate positive(80.80%) and weakly positive(14,14%),it didn’t not correlate well with the tube method for detecting biofilm formation in E.coli.The rates of antibiotic resistance of biofilm producing E.coli were found to be 100%for chloramphenicol and amoxyclav(amoxicillin and clavulanic acid),86%for gentamicin and cefotaxime.84%for ceftazidime,83%for cotrimoxazole and piperacillin/tazobactam,75%for tetracycline and 70% for amikacin,Conclusions:This study reveals the prevalence and antimicrobial susceptibility pattern of biofilm and non-biofilm producing uropathogenic E.coli strains.

Progress toward understanding the contribution of alkali generation in dental biofilms to inhibition of dental caries

Alkali production by oral bacteria is believed to have a major impact on oral microbial ecology and to be inibitory to the initiation and progression of dental caries. A substantial body of evidence is beginning to accumulate that indicates the modulation of the alkalinogenic potential of dental biofilms may be a promising strategy for caries control. This brief review highlights recent progress toward understanding molecular genetic and physiologic aspects of important alkali-generating pathways in oral bacteria, and the role of alkali production in the ecology of dental biofilms in health and disease.

Biofilms and Helicobacter pylori: Dissemination and persistence within the environment and host

The presence of viable Helicobacter pylori(H. pylori) in the environment is considered to contribute to the levels of H. pylori found in the human population, which also aids to increase its genetic variability and its environment adaptability and persistence. H. pylori form biofilms both within the in vitro and in vivo envi-ronment. This represents an important attribute that assists the survival of this bacterium within environ-ments that are both hostile and adverse to prolifera-tion. It is the aim of this paper to review the ability of H. pylori to form biofilms in vivo and in vitro and to address the inherent mechanisms considered to sig-nificantly enhance its persistence within the host and in external environments. Furthermore, the dissemi-nation of H. pylori in the external environment and within in the human body and its impact upon infec-tion control shall be discussed.

Targeted Antimicrobial Therapy Against Streptococcus mutans Establishes Protective Non-cariogenic Oral Biofilms and Reduces Subsequent Infection

Aim Dental biofilms are complex communities composed largely of harmless bacteria. Certain pathogenic species including Streptococcus mutans （S. mutans） can become predominant when host factors such as dietary sucrose intake imbalance the biofilm ecology. Current approaches to control S. mutans infection are not pathogen-specific and eliminate the entire oral community along with any protective benefits provided. Here, we tested the hypothesis that removal of S. mutans from the oral community through targeted antimicrobial therapy achieves protection against subsequent S. mutans colonization. Methodology Controlled amounts of S. mutans were mixed with S. mutans-free saliva, grown into biofilms and visualized by antibody staining and cfu quantization. Two specifically-targeted antimicrobial peptides （STAMPs） against S. mutans were tested for their ability to reduce S. mutans biofilm incorporation upon treatment of the inocula. The resulting biofilms were also evaluated for their ability to resist subsequent exogenous S. mutans colonization. Results S. mutans colonization was considerably reduced （9 ± 0.4 fold reduction, P=0.01） when the surface was preoccupied with saliva-derived biofilms. Furthermore, treatment with S. mutans-specific STAMPs yielded S. mutans-deficient biofilms with significant protection against further S. mutans colonization （5 minutes treatment： 38 ± 13 fold reduction P=0.01; 16 hours treatment： 96 ± 28 fold reduction P=0.07）. Conclusion S. mutans infection is reduced by the pre- sence of existing biofilms. Thus maintaining a healthy or ＂normal＂ biofilm through targeted antimicrobial therapy （such as the STAMPs） could represent an effective strategy for the treatment and prevention of S. mutans colonization in the oral cavity and caries progression.

Biofilm three-dimensional architecture influences in situ pH distribution pattern on the human enamel surface

To investigate how the biofilm three-dimensional(3D) architecture influences in situ pH distribution patterns on the enamel surface. Biofilms were formed on human tooth enamel in the presence of 1% sucrose or 0.5% glucose plus 0.5% fructose. At specific time points, biofilms were exposed to a neutral pH buffer to mimic the buffering of saliva and subsequently pulsed with 1% glucose to induce re-acidification. Simultaneous 3D pH mapping and architecture of intact biofilms was performed using two-photon confocal microscopy. The enamel surface and mineral content characteristics were examined successively via optical profilometry and microradiography analyses. Sucrose-mediated biofilm formation created spatial heterogeneities manifested by complex networks of bacterial clusters(microcolonies). Acidic regions(pH<5.5) were found only in the interior of microcolonies,which impedes rapid neutralization(taking more than 120 min for neutralization). Glucose exposure rapidly re-created the acidic niches, indicating formation of diffusion barriers associated with microcolonies structure. Enamel demineralization(white spots),rougher surface, deeper lesion and more mineral loss appeared to be associated with the localization of these bacterial clusters at the biofilm-enamel interface. Similar 3D architecture was observed in plaque-biofilms formed in vivo in the presence of sucrose. The formation of complex 3D architectures creates spatially heterogeneous acidic microenvironments in close proximity of enamel surface, which might correlate with the localized pattern of the onset of carious lesions(white spot like) on teeth.

Comment on “Effect of biofilm formation by clinical isolates of Helicobacter pylori on the efflux-mediated resistance to commonly used antibiotics”

Attaran et al[1] have recently shown that decreased susceptibility of established Helicobacter pylori(H. pylori) biofilms to specific antibiotics,was associated with the overtly enhanced transcription of two efflux pump genes,hp1165 and hef A,involved in specific resistance to tetracycline and multiple antibiotics,respectively. Apart from antibiotic exposure,secretion of multiple antimicrobial peptides,such as human β-defensins(hβDs),by the gastric epithelium upon Hp challenge,may act as early triggering events that positively impact biofilm formation and thus,antibiotic resistance. In this regard,we undertook genomic transcriptional studies using Hp 26695 strain following exposure to sublethal,similar to those present in the gastric niche,concentrations of hβDs in an attempt to provide preliminary data regarding possible mechanisms of immune evasion and selective sensitivity of Hp. Our preliminary results indicate that hβD exposure ignites a rapid response that is largely due to the activation of several,possibly interconnected transcriptional regulatory networks – origons-that ultimately coordinate cellular processes needed to maintain homeostasis and successful adaptation of the bacterium in the gastric environment. In addition,we have shown that both antibiotic and hβD resistance are mediated by dedicated periplasmic transporters,including the aforementioned efflux pump genes hp1165 and hef A,involved in active export of antibiotics from the cell membrane and/or,as recently suggested,substrate sensing and signalling. Furthermore,itappears that sublethal doses of hβDs may enhance biofilm formation by the sustained expression of,mainly,quorum sensing-related genes. In conclusion,we provide additional data regarding the role of specific innate immune molecules in antibiotic cross-resistance mechanisms that may deepen our understanding in the context of the development of novel eradication regimens.

Insights into Cronobacter sakazakii Biofilm Formation and Control Strategies in the Food Industry

Cronobacter sakazakii(C.sakazakii)is a foodborne opportunistic pathogen that can cause life-threatening invasive diseases,such as necrotizing enterocolitis,meningitis,and sepsis in infants.The potential risk of C.sakazakii contamination of powdered infant formula(PIF)is an issue that has attracted considerable attention from manufacturers,regulators,and consumers.C.sakazakii biofilms on the surfaces of equipment and in diverse food-production environments constitute a mode of cell growth that protects the pathogen from hostile environments,and are an important source of persistent contamination of food products.Bacterial biofilms are difficult to remove due to their resistant properties.Conventional cleaning and sterilizing procedures may be insufficient for biofilm control,and may lead to further biofilm development and dispersal.Consequently,novel control strategies are being developed,such as nanotechnology-based delivery systems,interspecies interactions,antimicrobial molecules of microbial origin,natural extracts,and phages.This review focuses on describing the mechanisms underlying the biofilm formation and behavior of C.sakazakii and discussing potential control strategies.

The photodynamic therapy on Streptococcus mutans biofilms using erythrosine and dental halogen curing unit

The purpose of our study was to evaluate the effect of photodynamic therapy （PDT）, using erythrosine as a photosensitizing agent and a dental halogen curing unit as a light source, on Streptococcus mutans in a biofilm phase. The S. mutans biofilms were formed in a 24-well cell culture cluster. Test groups consisted of biofilms divided into four groups： group 1： no photosensitizer or light irradiation treatment （control group）; group 2： photosensitizer treatment alone; group 3： light irradiation alone; group 4： photosensitizer treatment and light irradiation. After treatments, the numbers of colony-forming unit （CFU） were counted and samples were examined by confocal laser scanning fluorescence microscopy （CLSM）. Only group 4 （combined treatment） resulted in significant increases in cell death, with rates of 75% and 55% after 8 h of incubation, and 74% and 42% at 12 h, for biofilms formed in brain-heart infusion （BHI） broth supplemented with 0% or O. 1% sucrose, respectively. Therefore, PDT of S. mutans biofilms using a combination of erythrosine and a dental halogen curing unit, both widely used in dental clinics, resulted in a significant increase in cell death. The PDT effects are decreased in biofilms that form in the presence of sucrose.