Inhibitory effect of isolated Lactobacillus from oral cavity against bacterial Pathogens and its effect on health promotion Efecto inhibidor de Lactobacillus aislado de la cavidad oral contra patógenos bacte- rianos y su efecto en la promoción de la salud Khalil Azizian1,2, 3, Sepehr Taghizadeh1, Rasoul Hosseinpour3, Asghar Tanomand4, Elham Sheykhsaran1, Naser Alizadeh2, Behrooz Naghili1, Hossein Samadi Kafil1,* 1. Infectious and Tropical Diseases Research Center, Tabriz university of medical science, Tabriz, Iran. 2. Department of Medical Microbiology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran. 3. Liver and Gastrointestinal Diseases Research Center, Tabriz university of medical science, Tabriz, Iran. 4. Department of Basic Sciences, Maragheh University of Medical Sciences, Maragheh, Iran. http://dx.doi.org/10.30827/ars.v60i1.7943 Artículo original ABSTRACT Original Article Aims: To determine the inhibitory effect of oral Lactobacillus against bacterial pathogens and investigate Correspondencia correlation between presence of Lactobacillus strains and health promotion. Correspondence Hossein Samadi Kafil Method: One hundred saliva samples were collected from oral cavity of domestic dairy consumers and PhD, Assistant professor were investigated for the isolation and identification of Lactobacillus strain by conventional culture and Department of Microbiology, Faculty of sequencing of 16SrRNA. Furthermore, well diffusion assay was performed to determination of antibac- Medicine, Tabriz University of Medical terial activity of Lactobacillus strains against bacterial pathogens including Salmonella typhimurium, Kleb- Sciences, Tabriz, Iran [email protected] siella pneumoniae, Shigella sonnei, Shigella dysenteriae, Enterococcus faecalis and Enterococcus faecium. Finally, +98-9127184735 association between health condition and isolation of Lactobacillus were investigated and obtained data Financiación using questionary form were analysed by chi-square test. Fundings Results: Thirty Lactobacillus strains recovered from 100 hundred saliva samples. The The authors declare that no funding whatsoever was received from any sour- most common isolated strain was L. gasseri (n=18) and followed by L. vaginalis (n=3) and ce in carrying out the research work. L. salivarius (n=3). All Lactobacillus strains demonstrated antibacterial activity against at Agradecimientos least one of the investigated pathogens. However, the strongest results were obtained by L. vaginalis Acknowledgements against K. pneumonia. The correlation between the presence of thirty Lactobacillus strains and health pro- This study was granted by Infectious motion not found. However, only L. gasseri species has significant positive impact on health in their hosts and Tropical Diseases Research Center, (P < 0.05). Tabriz University of Medical Sciences and approved by Tabriz University of Conclusion: Only some Lactobacillus species have a positive impact on health promotion. Despite of Medical Sciences. Authors would like to weak activity against the investigated pathogens, L. gasseri has a positive impact on the mental problem thank all staff of Biomedicine (Medical (intense anger and depression) of their hosts. Biology) Research Institute. This study was supported by Infectious Keywords: Lactobacillus; Health condition; Antibacterial activity; Oral cavity. and Tropical Diseases Research Center, Tabriz University of Medical Sciences. We would like to thank all participants ABSTRACT and DARC center for their collaboration. Objetivos: Determinar el efecto inhibidor del Lactobacillus aislado de la cavidad oral contra patógenos Conflicto de interés bacterianos e investigar la correlación entre la presencia de cepas de Lactobacillus y la promoción de la Competing interest salud. All Authors declare no conflicts of interest. Método: se recolectaron cien muestras de saliva de la cavidad oral de consumidores de productos lácteos Declaración ética y se investigó el aislamiento e identificación de la cepa de Lactobacillus mediante cultivo convencional y Ethical Statement secuenciación de 16SrRNA. Además, se realizó un ensayo de difusión en pocillos para determinar la ac- This study has no results of experimen- tividad antibacteriana de las cepas de Lactobacillus contra patógenos bacterianos que incluyen Salmonella tal studies on human participants or typhimurium, Klebsiella pneumoniae, Shigella sonnei, Shigella dysenteriae, Enterococcus faecalis y Enterococcus use of laboratory animals, therefore no Ethical approve is required. faecium. Finalmente, se investigó la asociación entre el estado de salud y el aislamiento de Lactobacillus y se obtuvieron los datos utilizando el cuestionario mediante la prueba de chi-cuadrado. Received: 15.09.2018 Accepted: 08.02.2019 Resultados: Se aislaron treinta cepas de Lactobacillus de 100 muestras de saliva. La cepa aislada más común fue L. gasseri (n = 18), seguida por L. vaginalis (n = 3) y L. salivarius (n = 3). Todas las cepas de LICENSE 3.0 UNPORTED. Lactobacillus demostraron actividad antibacteriana contra al menos uno de los patógenos investigados. Ars Pharm. 2019; 60(1): 27-33 27 Azizian K, et al. Sin embargo, los resultados más fuertes fueron obtenidos por L. have useful role in promotion of oral and gastrointestinal vaginalis contra K. pneumonia. No se encontraron correlación entre health 4. la presencia de algunas de las treinta cepas de Lactobacillus y la promoción de la salud. Sin embargo, solo la especie L. gasseri tuvo The milk products including yogurt and cheese are im- un impacto positivo significativo en la salud de sus hospedadores portant dietary supplements which contain Lactobacillus (P <0,05). 14. Consumption of these products is main way to probi- Conclusión: solo algunas especies de Lactobacillus tienen un im- otic transmission to the host body. Thus, there is probably pacto positivo en la promoción de la salud. A pesar de la actividad strong association between diet and commensal microor- débil contra los patógenos investigados, L. gasseri tiene un impacto ganism including probiotics bacteria 15. positivo en el problema mental (ira intensa y depresión) de sus huéspedes. The aims of this study were investigate the inhibitory activ- ity of isolated Lactobacillus from oral cavity and also, deter- Palabras clave: Lactobacillus; condición de salud; actividad antibac- mination of association between isolation rates of Lactoba- teriana; cavidad oral. cillus and health quality in the host. INTRODUCTION MATERIAL AND METHODS In the past decade role of probiotic microorganisms in Study population health promotion and as preventative agents were inves- 1,2 tigated . Probiotics described as microorganisms which One hundred healthy persons who use domestic dairy 3,4 are useful if used in sufficient number . Different bacteria products were recruited in this study. The study was con- such as Bifidobacterium, Streptococcus, and Lactobacillus in- ducted from May to August 2017 at rural area of Bukan, troduced as probiotics, and known as generally recognized Western Azerbaijan province, Iran. Also, the questioner 3,5,6 as safe (GRAS) and non-pathogenic bacteria . Recently, forms were applied to all persons. These forms were in- Lactobacillus spp. considered as main probiotic in medical cluded age, gender, experience of infectious disease, severe 1,2 and industrial scope . Lactobacillus spp. are heterogeneous gastric pain, herpes, and mental problem (intense anger group of bacteria including about 145 species of non-spore and depression). The current study was approved by the forming and Gram-positive rods which can be grown in Research Ethics Committee of Tabriz University of Medical 3 anaerobic or microaerophilic conditions . Also, Lactobacil- Science; also signed informed consent form was obtained lus was used in dairy and fermented food for promotion from each patient prior to the initiation of sampling. Par- 3 of health quality . However, Lactobacillus spp. is presented ticipants were subjected for sampling without any haz- in various places like environment and human body (oral ardous action and participation was completely volunteer. 1,2,5,7 cavity, gastrointestinal and vaginal tract) . This research has been conducted in full accordance with the World Medical Association Declaration of Helsinki and It was previously demonstrated that consumption of Lacto- was done after confirmation of ethic committee of Tabriz bacillus has several benefits including decrease of time and University of medical sciences. severity of diarrhea 3, antibacterial and antifungal charac- 8 7 teristics , and improvement of vaginal health . Sample collection Inhibitory activity against the pathogens is one of the most According to Kohler and Bratthall (1979), sample collec- characteristics of bacteria which known as probiotics. Pro- tion was performed by modified spatula method 16. Briefly, biotic inhibitory activity is related to reduce of pathogenic wooden spatula was inserted in oral cavity to moisten it colonization and causing adverse growth conditions for with saliva. Then both sides of spatula placed on Rogosa SL pathogens by production of hydrogen peroxide (H2O2) agar (MicroMaster, India). The plates were incubated under and bacteriocin 8-10. Moreover, it has recently been demon- anaerobic condition (80% N2, 10% H2, and 10% CO2) pro- strated that Lactobacillus species isolated from human are vided by Anoxomat (Mart Microbiology BV) at 37°C for 72 better probiotics, because these strains were adapted to the h. Then dominant