Abstract
Background: Probiotics are living microorganisms that have a healthy influence on a host.
Objective: The aim of this study was to isolate a probiotic Pediococcus acidilactici strain from newborn faeces and develop and optimize a selective high-performance liquid chromatography method for the determination and validation of its lactic acid content and also evaluate some probiotic characteristics.
Methods: Isolated strains were identified by the API 50 CH system and 16S rDNA gene sequence analysis and tested for antibiotic susceptibility, bile salt tolerance, low pH resistance, proteolytic, haemolytic activity, as well as the production of bacteriocin, hydrogen peroxide, and lactic acid. Antimicrobial activity of selected strain against standard test microorganisms was determined by the spot lawn method and the quantitation of lactic acid was carried out by high-performance liquid chromatography on a Rezex ROA organic acid (300x7.8 mm) analytical column.
Results: P. acidilactici M7 strain was evaluated as a potential probiotic due to its ability to survive at low pH values or in the presence of pepsin, pancreatin, and bile salts. The lactic acid amount of strain was found in the range between 5.59-5.94 mg mL-1 by HPLC. M7 strain was also found to be resistant to vancomycin, had no bacteriocin, and hydrogen peroxide production and was able to inhibit the growth of P. aeruginosa and E. faecalis by its lactic acid content.
Conclusion: This study explains a simple, selective, and fully validated procedure for the determination of lactic acid from probiotic bacteria.
Keywords: HPLC, lactic acid, Pediococcus acidilactici, probiotic ability, quantification, validation.
Graphical Abstract
[2]
Redondo-Lopez, V.; Cook, R.L.; Sobel, J.D. Emerging role of lactobacilli in the control and maintenance of the vaginal bacterial microflora. Rev. Infect. Dis., 1990, 12, 856-872.
[3]
Gumustas, M.; Uslu, B.; Ozkan, S.A. In:Soft chemistry and food fermentation; Grumezescu, A.M.; Holban, A.M., Eds.; Elsevier: Netherlands, 2017.
[4]
Klare, I.; Konstabel, C.; Werner, G.; Huys, G.; Vankerckhoven, V.; Kahlmeter, G.; Hildebrandt, B.; Müller-Bertling, S.; Witte, W.; Goossens, H. Antimicrobial susceptibilities of Lactobacillus, Pediococcus and Lactococcus human isolates and cultures intended for probiotic or nutritional use. J. Antimicrob. Chemother., 2007, 59, 900-912.
[5]
Papagianni, M.; Anastasiadou, S. Encapsulation of Pediococcus acidilactici cells in corn and olive oil microcapsules emulsified by peptides and stabilized with xanthan in oil-in-water emulsions: studies on cell viability under gastro-intestinal simulating conditions. Enzyme Microb. Technol., 2009, 45, 514-522.
[6]
Okorhi, B.F. Anti-pseudomonas activity of organic acids produced by lactic acid bacteria. IBSPR, 2014, 2, 106-114.
[7]
Ray, B.; Bhunia, A. Fundamental food microbiology; 5th ed.; CRC Press, Taylor & Francis Group: Boca Raton,. , 2013.
[8]
Verna, E.C.; Lucak, S. Use of probiotics in gastrointestinal disorders: What to recommend. Therap. Adv. Gastroenterol., 2010, 3, 307-319.
[9]
Anal, A.K.; Singh, H. Recent advances in microencapsulation of probiotics for industrial applications and targeted delivery. Trends Food Sci. Technol., 2007, 18, 240-251.
[10]
Elmer, G.W.; Surawicz, C.M.; Mcfarland, L.V. Biotherapeutic agents: A negleeted modality for the treatment and prevention of selected intestinal and vaginal infections. JAMA, 1996, 275, 870-876.
[11]
Gan, B.S.; Kim, J.; Reid, G.; Cadieux, P.; Howard, J.C. Lactobacillus fermentum RC-14 inhibits Staphylococcus aureus infection of surgical implants in rats. J. Infect. Dis., 2002, 185, 1369-1372.
[12]
Giraffa, G.; Carminati, D. In: Molecular techniques in the microbial ecology of fermented foods; Cocolin, L.; Ercolini, D., Ed.; Springer Science + Business Media: New York, . , 2008.
[13]
Gumustas, M.; Kurbanoglu, S.; Uslu, B.; Ozkan, S.A. UPLC versus HPLC on drug analysis: Advantageous, applications and their validation parameters. Chromatographia, 2013, 76, 1365-1427.
[14]
Guetarni, H.; Bensaid, A.; Bensoltane, A. Analysis of lactic acid responsible for inhibition in-vitro of Helicobacter pylori by high performance chromatography (HPLC). J. Biotechnol. Lett., 2012, 3, 34-36.
[15]
Özçelik, S. Organic acid production by lactic acid bacteria in different fish infusion broths using HPLC., MSD Thesis, Çukurova University: Adana, . 2011.
[16]
Yang, V.A.; Clausen, C.A. Antifungal metabolites of Lactobacilli.In:Woodframe Housing Durability and Disaster Issues Conference, Las Vegas, Nevada, USA Madison, WI,2004, pp. 307-311.
[17]
Kordiš-Krapež, M.; Abram, V.; Kač, M.; Ferjančič, S. Determination of organic acids in white wine by RP-HPLC. Food Technol. Biotechnol., 2001, 39, 93-99.
[18]
ICH., Harmonised Tripartite Guideline Validation of Analytical Procedure Text and Methodology Q2 (R1); International Council on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use, 2005.
[20]
Blaiotta, G.; Pepe, O.; Mauiello, G.; Villani, F.; Andolfi, R.; Moschetti, G. 16S-23S rDNA intergenic spacer region polymorphism of Lactococcus garvieae, Lactococcus raffinolactis and Lactococcus lactis as revealed by PCR and nucleotide sequence analysis. Syst. Appl. Microbiol., 2002, 25, 520-527.
[21]
European Food Safety Authority (EFSA).Guidance on the assessment of bacterial susceptibility of antimicrobials of human and veterinary importance: EFSA Panels on additives and products or substances used in animal feed. EFSA J., 2012, 10, 2740.
[22]
Maragkoudakis, P.A.; Zoumpopoulou, G.; Miaris, C.; Kalantzopoulos, G.; Pot, B.; Tsakalidou, E. Probiotic potential of Lactobacillus strains isolated from dairy products. Int. Dairy J., 2006, 16, 189-199.
[23]
Charteris, W.P.; Kelly, P.M.; Morelli, L.; Collins, J.K. Development of an in vitro methodology to determine the transit tolerance of potentially probiotic Lactobacillus and Bifidobacterium species in the upper human gastrointestinal tract. J. Appl. Microbiol., 1998, 84, 759-768.
[24]
Lima, E.T.; Andreatti Filho, R.L.; Okamoto, A.S.; Noujaim, J.C.; Barros, M.R.; Crocci, A.J. Evaluation in vitro of the antagonistic substances produced by Lactobacillus spp. isolated from chickens. Can. J. Vet. Res., 2007, 71, 103-107.
[25]
Sahnouni, F.; Boutiba-Maatallah, A.; Bouhadi, D.; Boutiba, Z. Characterization of bacteriocin produced by Lactococcus lactis ssp. lactis strains isolated from marine fish caught in Algerian west coast. TURKJANS, 2014, 2, 1838-1843.
[26]
Tuncer, Y.; Özden, B.; Avşaroğlu, M.D. Determination of some microbiological characteristics and antibacterial activity of lactic acid bacteria isolates of boza. Süleyman Demirel Uni. Physical Sci. Inst. J., 2008, 12, 19-25.
[27]
Ozkan, S.A.; Kauffmann, J.M.; Zuman, P. In:Biomedical and pharmaceutical sciences: voltammetry, amperometry, biosensors, applications; Springer: Berlin, 2015.
[28]
Waters, V.; Smyth, A. Cystic fibrosis microbiology: Advances in antimicrobial therapy. J. Cyst. Fibros., 2015, 14, 551-560.
[29]
Food and Agriculture Organization/World Health Organization. Guidelines for the evaluation of probiotics in food: report of a joint FAO/WHO working group on drafting guidelines for the evaluation of probiotics in food. London, , 2002.
[30]
Abbasiliasi, S.; Tan, J.S.; Ibrahim, T.A.T.; Ramanan, R.T.; Vakhshiteh, F.; Mustafa, S.; Ling, T.C.; Rahim, R.A.; Ariff, A.B. Isolation of Pediococcus acidilactici Kp10 with ability to secrete bacteriocin-like inhibitory substance from milk products for applications in food industry. BMC Microbiol., 2012, 12, 260-271.
[31]
Barbosa, J.; Borges, S.; Teixeira, P. Pediococcus acidilactici as a potential probiotic to be used in food industry. Int. J. Food Sci. Technol., 2015, 50, 1151-1157.
[32]
Osmanagaoglu, O.; Kiran, F.; Ataoglu, H. Evaluation of in vitro probiotic potential of Pediococcus pentosaceus OZF isolated from human breast milk. Probiotics Antimicrob. Proteins, 2010, 2, 162-174.
[33]
Haakensen, M.; Vickers, D.M.; Ziola, B. Susceptibility of Pediococcus isolates to antimicrobial compounds in relation to hop-resistance and beer-spoilage. BMC Microbiol., 2009, 9, 190.
[34]
Loomis, H.F. Digestion in the Stomach., Food Enzyme Institute. http://www.foodenzymeinstitute.com/articles/digestion-in-the-stomach.aspx?list=bydate&print=yes. (Accessed October 8, 2016).
[35]
Feng, J.; Wang, L.; Zhou, L.; Yang, X.; Zhao, X. Using in vitro immunomodulatory properties of lactic acid bacteria for selection of probiotics against salmonella infection in broiler chicks. PLoS One, 2016, 11, 1-14.
[36]
Noohi, N.; Ebrahimipour, G.; Rohani, M.; Talebi, M.; Pourshafie, M.R. Evaluation of potential probiotic characteristics and antibacterial effects of strains of Pediococcus species isolated from broiler chickens. Br. Poult. Sci., 2016, 57, 317-323.
[37]
Oh, Y.J.; Jung, D.S. Evaluation of probiotic properties of Lactobacillus and Pediococcus strains isolated from Omegisool, a traditionally fermented millet alcoholic beverage in Korea. LWT-Food Sci. Technol., 2015, 63, 437-444.
[38]
Begley, M.; Sleator, R.D.; Gahan, C.G.M.; Hill, C. The contribution of three bileassociated loci (bsh, pva, btlB) to gastrointestinal persistence and bile tolerance of Listeria monocytogenes. Infect. Immun., 2015, 73, 894-904.
[39]
Kaboré, D.; Sawadogo-Lingani, H.; Dicko, M.H.; Diawara, B.; Jakobsen, M. Acid resistance, bile tolerance and antimicrobial properties of dominant lactic acid bacteria isolated from traditional “maari” baobab seeds fermented condiment. Afr. J. Biotechnol., 2012, 11, 1197-1206.
[40]
Servin, A.L. Antagonistic activities of lactobacilli and bifidobacteria against microbial pathogens. FEMS Microbiol. Rev., 2004, 28, 405-440.
[41]
Ibrahim, S.A.; Yang, H.; Seo, C.W. Antimicrobial activity of lactic acid and copper on growth of Salmonella and Escherichia coli O157:H7 in laboratory medium and carrot juice. Food Chem., 2008, 109, 137-143.
Article Metrics
37
3
1
1