Isolation of the Bacteriophages Inhibiting the Expression of the Genes Involved in Biofilm Formation by Streptococcus mutans

Background: The potential of Streptococcus mutans for biofilm formation makes it one of the main organisms causing dental caries. Various preventive strategies have been applied to reduce tooth decay. Objectives: In the current study, we aimed to isolate S. mutans bacteriophages from sewage and to investigate their effects on the expression of the genes involved in bacterial biofilm formation in dental caries. Methods: Eighty-one dental plaque samples were collected. Then to isolate and identify S. mutans, bacterial culture media and molecular tests were used. Moreover, the biofilm formation capability of the isolated S. mutans was determined. Also, lytic bacteriophages were isolated from raw urban sewage, and phage morphology was determined by transmission electron microscopy (TEM). Real-time PCR was used to assess the effects of the isolated bacteriophages on the expression of the genes involved in biofilm formation. Results: Overall, 32 (39.5%) samples were positive for the presence of S. mutans. All of the isolates contained the gtfD gene. The frequencies of other genes were as follows: gtfB (17, 53.12%), gtfC (19, 53.37%), SpaP (13, 40.62%), and luxS (23, 17.87%). The isolated S. mutans bacteria presented different ranges of biofilm formation ability. Based on TEM results, two sewage-isolated bacteriophages, belonging to Siphoviridae and Tectiviridae families, were able to prevent biofilm formation up to 97%. Conclusions: Our findings indicate that phage therapy can be an optional way for controlling biofilm development and reducing the colonization of teeth surface by S. mutans.

[1]  S. Palmer,et al.  The Biology of Streptococcus mutans , 2019, Microbiology spectrum.

[2]  D. Drake,et al.  Are the mutans streptococci still considered relevant to understanding the microbial etiology of dental caries? , 2018, BMC Oral Health.

[3]  E. Bair,et al.  The oral bacterial microbiome of occlusal surfaces in children and its association with diet and caries , 2017, PloS one.

[4]  A. Rahimi Foroushani,et al.  Phage therapy: assessment of the efficacy of a bacteriophage isolated in the treatment of salmonellosis induced by Salmonella enteritidis in mice , 2017, Gastroenterology and hepatology from bed to bench.

[5]  K. Gregorczyk,et al.  Phage Therapy in Bacterial Infections Treatment: One Hundred Years After the Discovery of Bacteriophages , 2016, Current Microbiology.

[6]  Fabien J. Cousin,et al.  Isolation of a Novel Phage with Activity against Streptococcus mutans Biofilms , 2015, PloS one.

[7]  Chengfei Zhang,et al.  Morphological and proteomic analyses of the biofilms generated by Streptococcus mutans isolated from caries-active and caries-free adults , 2015 .

[8]  M. Bouzari,et al.  Biotechnological Applications of Two Novel Lytic Bacteriophages of Streptococcus mutans in Tooth Decay Bio-Controlling , 2015 .

[9]  A. Mira,et al.  Solving the etiology of dental caries. , 2015, Trends in microbiology.

[10]  L. Jianjun,et al.  Effect of Lactobacillus species on Streptococcus mutans biofilm formation. , 2014, Pakistan journal of pharmaceutical sciences.

[11]  S. Gharavi,et al.  Assessment of Biofilm Formation and Resistance to Imipenem and Ciprofloxacin among Clinical Isolates of Acinetobacter baumannii in Tehran , 2014, Jundishapur journal of microbiology.

[12]  S. Opal,et al.  A historical overview of bacteriophage therapy as an alternative to antibiotics for the treatment of bacterial pathogens , 2013, Virulence.

[13]  E. Ferdosi,et al.  Frequency, biofilm formation and acid susceptibility of streptococcus mutans and streptococcus sobrinus in saliva of preschool children with different levels of caries activity , 2013, Dental research journal.

[14]  S. Kazem,et al.  The Role Of Streptococcus Mutants In Dental Caries In Two Groups Of Sensitive And Resistance Children Age Between 3 To 5 Years , 2013 .

[15]  M. Okada,et al.  Caries prevalence associated with Streptococcus mutans and Streptococcus sobrinus in Japanese schoolchildren. , 2012, International journal of paediatric dentistry.

[16]  R. Salehi,et al.  Lactobacillus Acidophilus-Derived Biosurfactant Effect on GTFB and GTFC Expression Level in Streptococcus Mutans Biofilm Cells , 2011, Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology].

[17]  M. Kutateladze,et al.  Bacteriophages as potential new therapeutics to replace or supplement antibiotics. , 2010, Trends in biotechnology.

[18]  M. Björklund,et al.  Streptococcus mutans, Caries and Simulation Models , 2010, Nutrients.

[19]  M. Jalali,et al.  The effect of nisin on biofilm forming foodborne bacteria using microtiter plate method , 2009 .

[20]  F. A. de Ávila,et al.  Detection of Streptococcus mutans and Streptococcus sobrinus in dental plaque samples from Brazilian preschool children by polymerase chain reaction. , 2007, Brazilian dental journal.

[21]  M. Shemesh,et al.  Expression of biofilm-associated genes of Streptococcus mutans in response to glucose and sucrose. , 2007, Journal of medical microbiology.

[22]  P. Taylor,et al.  Isolation of bacteriophages from the oral cavity , 2004, Letters in applied microbiology.

[23]  T. Fujiwara,et al.  PCR detection and identification of oral streptococci in saliva samples using gtf genes. , 2004, Diagnostic microbiology and infectious disease.

[24]  D. Steinberg,et al.  Bacteriophage isolation from human saliva , 2003, Letters in applied microbiology.

[25]  M. Gabriele,et al.  Effect of removable orthodontic appliances on oral colonisation by mutans streptococci in children. , 2001, European journal of oral sciences.

[26]  W. Summers,et al.  Bacteriophage therapy. , 2001, Annual review of microbiology.

[27]  K. Hirota,et al.  Detection of Streptococcus mutans by PCR amplification of spaP gene. , 1994, Journal of medical microbiology.