Microbiological Quality of Probiotic Products

Gordana Zavišić; Slavica Ristić; Branka Petković; Dragoslava Živkov Šaponja; Nikola Jojić; Drina Janković

doi:10.5937/arhfarm73-42160

Gordana Zavišić University Business Academy in Novi Sad, Faculty of Pharmacy
Slavica Ristić University of Belgrade – Faculty of Medicine, Institute for Medical and Clinical Biochemistry http://orcid.org/0000-0002-6983-4412
Branka Petković University of Belgrade – Institute for Biological Research “Siniša Stanković” – National Institute of the Republic of Serbia, Department of Neurophysiology http://orcid.org/0000-0001-7817-4092
Dragoslava Živkov Šaponja University Business Academy in Novi Sad, Faculty of Pharmacy
Nikola Jojić University Business Academy in Novi Sad, Faculty of Pharmacy https://orcid.org/0000-0002-9926-7658
Drina Janković University of Belgrade, Vinča Institute of Nuclear Sciences – National Institute of the Republic of Serbia

DOI: https://doi.org/10.5937/arhfarm73-42160

Keywords: probiotic, microbiological quality, safety, stability

Abstract

Microorganisms used as probiotics should meet elementary safety aspects (non-toxicity, absence of antibiotic resistance genes and translocation) and functional/technological aspects (resistance and survival in the acid gastric environment, adhesiveness, stability, and cell viability). Probiotics with the health claim of being a dietary product or a pharmabiotic (drug category) should be clinically tested, validated, documented, and continuously controlled for quality. Important quality parameters include the identification of declared probiotic strains, the number of viable microorganisms (probiotic bacteria and/or fungi), and microbiological purity (absence of specified pathogenic/opportunistic pathogenic bacteria and fungi, and limitation of total unspecified contaminants such as aerobic bacteria, yeasts, and molds). Due to numerous reports of low-quality commercial probiotics marketed for human use, this review discusses the methods used to test the probiotic microorganism content, safety for the intended use, and proven health benefits of those probiotics whose microbiological quality deviates from the manufacturer’s stated content, as well as the maintenance of cell viability, i.e., stability of the probiotic during the shelf life. In addition, the adverse effects of probiotics and the potential hazards to the health of the user are addressed.

References

1. Food and Agriculture Organization/World Health Organisation. Evaluation of Health and Nutritional Properties of Probiotics in Food. Córdoba, 2001.

2. Hill C, Guarner F, Reid G, Gibson GR, Merenstein DJ, Pot B, et al. Expert consensus document. The International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nat Rev Gastroenterol Hepatol. 2014;11(8):506-14.

3. Kong Q, Liu T, Xiao H. Editorial: Effects of probiotics and prebiotics on gut pathogens and toxins. Front Microbiol. 2022;13:856779.

4. Zawistowska-Rojek A, Zaręba T, Tyski S. Microbiological testing of probiotic preparations. Int J Environ Res Public Health. 2022;19(9):5701.

5. Patra F, Duary RK. Determination and safety aspects of probiotic cultures. In: Panesar PS, Anal AK, editors. Probiotics, Prebiotics and Synbiotics. New York: John Wiley & Sons, Inc.; 2022; p. 122-60.

6. Suvorov A. What is wrong with enterococcal probiotics? Probiotics Antimicrob Proteins. 2020;12(1):1-4.

7. EFSA on Dietetic Products, Nutrition and Allergies (NDA). Guidance on the scientific requirements for health claims related to gut and immune function. EFSA Journal. 2011;9(4):1984.

8. EFSA Panel on Dietetic Products, Nutrition, and Allergies (NDA). Scientific Opinion on Dietary reference values for water. EFSA Journal. 2010;8(3):1459.

9. EFSA Panel on Biological Hazards (BIOHAZ). Statement on the update of the list of QPS-recommended biological agents intentionally added to food or feed as notified to EFSA 14: suitability of taxonomic units notified to EFSA until March 2021. EFSA Journal. 2021;19(7):6689.

10. Binda S, Hill C, Johansen E, Obis D, Pot B, Sanders ME, et al. Criteria to qualify microorganisms as "probiotic" in foods and dietary supplements. Front Microbiol. 2020;11:1662.

11. Timmerman HM, Koning CJ, Mulder L, Rombouts FM, Beynen AC. Monostrain, multistrain and multispecies probiotics – A comparison of functionality and efficacy. Int J Food Microbiol. 2004;96(3):219-33.

12. Puvanasundram P, Chong CM, Sabri S, Yusoff MS, Karim M. Multi-strain probiotics: Functions, effectiveness and formulations for aquaculture applications. Aquac Rep. 2021;21:100905.

13. Arora M, Baldi A. Regulatory categories of probiotics across the globe: A review representing existing and recommended categorization. Indian J Med Microbiol. 2015;33(Suppl):2-10.

14. de Simone C. The unregulated probiotic market. Clin Gastroenterol Hepatol. 2019;17(5):809-17.

15. Saarela MH. Safety aspects of next generation probiotics. Curr Opin Food Sci. 2019;30:8-13.

16. Zavišić G, Radulović Ž, Vranić V, Begović J, Topisirović L, Strahinić I. Characterization and antimicrobial activity of vaginal Lactobacillus isolate. Arch Biol Sci. 2011;63(1):29-35.

17. Zavišić G, Petričević S, Radulović Ž, Begović J, Golić N, Topisirović L, et al. Probiotic features of two oral Lactobacillus isolates. Braz J Microbiol. 2012;43(1):418-28.

18. Prabhurajeshwar C, Chandrakanth RK. Probiotic potential of Lactobacilli with antagonistic activity against pathogenic strains: An in vitro validation for the production of inhibitory substances. Biomed J. 2017;40(5):270-83.

19. Lau LYJ, Chye FY. Antagonistic effects of Lactobacillus plantarum 0612 on the adhesion of selected foodborne enteropathogens in various colonic environments. Food Control. 2018;91:237-47.

20. Kim SJ, Park SH, Sin HS, Jang SH, Lee SW, Kim SY, et al. Hypocholesterolemic effects of probiotic mixture on diet-induced hypercholesterolemic rats. Nutrients. 2017;9(3):293.

21. Zavišić G, Ristić S, Petričević S, Novaković Jovanović J, Radulović Ž, Janać Petković B, et al. Characterisation and preliminary lipid-lowering evaluation of Lactobacillus isolated from a traditional Serbian dairy product. Benef Microbes. 2015;6(1):119-28.

22. Zavišić G, Ristić S, Rikalović M, Petković B, Janković D, Vukadinović A, Petričević S. Beneficial effects of probiotic supplementation on glucose and triglycerides in a mouse model of metabolic syndrome. J Funct Foods. 2022;95:105167.

23. de Almada CN, Almada CN, Martinez RC, Sant'Ana AS. Paraprobiotics: Evidences on their ability to modify biological responses, inactivation methods and perspectives on their application in foods. Trends Food Sci Technol. 2016;58:96-114.

24. Maria Carmen C, Seppo S, Vinderola G. Postbiotics: Defining the impact of inactivated microbes and their metabolites on promotion of health. In: Koren O, Rautava S, editors. The Human Microbiome in Early Life. Cambridge, CA, USA: Academic Press; 2021; p. 257-68.

25. Aggarwal S, Sabharwal V, Kaushik P, Joshi A, Aayushi A, Suri M. Postbiotics: From emerging concept to application. Front Sustain Food Syst. 2022;6:887642.

26. Thorakkattu P, Khanashyam AC, Shah K, Babu KS, Mundanat AS, Deliephan A, et al. Postbiotics: Current trends in food and pharmaceutical industry. Foods. 2022;11(19):3094.

27. Salminen S, Collado MC, Endo A, Hill C, Lebeer S, Quigley EMM, et al. The International Scientific Association of Probiotics and Prebiotics (ISAPP) consensus statement on the definition and scope of postbiotics. Nat Rev Gastroenterol Hepatol. 2021;18(9):649-67.

28. Żółkiewicz J, Marzec A, Ruszczyński M, Feleszko W. Postbiotics – A step beyond pre- and probiotics. Nutrients. 2020;12(8):2189.

29. Opinion of the Scientific Committee on a request from EFSA on the introduction of a Qualified Presumption of Safety (QPS) approach for assessment of selected microorganisms referred to EFSA. EFSA Journal. 2007;587:1-16.

30. U.S. Food and Drug Administration [Internet]. Generally Recognized as Safe (GRAS) [cited 2023 Feb 11]. Available from: http://www.fda.gov/Food/IngredientsPackagingLabeling/GRAS/default.htm.

31. Higgins JPT, Thomas J, Chandler J, Cumpston M, Li T, Page MJ, et al. Cochrane Handbook for Systematic Reviews of Interventions. 2nd Edition. Chichester (UK): John Wiley & Sons; 2019.

32. van Loveren H, Sanz Y, Salminen S. Health claims in Europe: probiotics and prebiotics as case examples. Annu Rev Food Sci Technol. 2012;3:247-61.

33. Cordaillat-Simmons M, Rouanet A, Pot B. Live biotherapeutic products: the importance of a defined regulatory framework. Exp Mol Med. 2020;52(9):1397-1406.

34. Hempel S, Newberry S, Ruelaz A, Wang Z, Miles JN, Suttorp MJ, et al. Safety of probiotics used to reduce risk and prevent or treat disease. Evid Rep Technol Assess (Full Rep). 2011;(200):1-645.

35. Salminen MK, Salminen M. Lactobacillus bacteremia, with special focus on the safety of probiotic Lactobacillus rhamnosus GG. Espoo: Minna K. Salminen; 2006. 95 p.

36. Berg RD. Bacterial translocation from the gastrointestinal tract. Adv Exp Med Biol. 1999;473:11-30.

37. Potruch A, Schwartz A, Ilan Y. The role of bacterial translocation in sepsis: a new target for therapy. Therap Adv Gastroenterol. 2022;15:17562848221094214.

38. Rubin IMC, Stevnsborg L, Mollerup S, Petersen AM, Pinholt M. Bacteraemia caused by Lactobacillus rhamnosus given as a probiotic in a patient with a central venous catheter: a WGS case report. Infect Prev Pract. 2022;4(1):100200.

39. Rossi F, Amadoro C, Colavita G. Members of the Lactobacillus Genus Complex (LGC) as opportunistic pathogens: A review. Microorganisms. 2019;7(5):126.

40. Kothari D, Patel S, Kim SK. Probiotic supplements might not be universally-effective and safe: A review. Biomed Pharmacother. 2019;111:537-47.

41. Pravilnik o dodacima ishrani (dijetetski suplementi). Službeni glasnik RS 45/2022.

42. European Pharmacopoeia. 10th ed., Strasbourg, France: EDQM, 2019; 876-878.

43. European Pharmacopoeia. 10th ed., Strasbourg, France: EDQM, 2019; 250-254.

44. European Pharmacopoeia. 10th ed., Strasbourg, France: EDQM, 2019; 255-258.

45. Dreher-Lesnick SM, Schreier JE, Stibitz S. Development of phage lysin LysA2 for use in improved purity assays for live biotherapeutic products. Viruses. 2015;7(12):6675-88.

46. United States Pharmacopeia. General Tests and Assays: Probiotic Tests, USP42-NF37, Rockville, MD, USA: United States Pharmacopeial Convention, 2019.

47. United States Pharmacopeia. General Chapter: Microbial Enumeration Tests –Nutritional and Dietary Supplements, USP42-NF37, Rockville, MD, USA: United States Pharmacopeia, 2019.

48. United States Pharmacopeia. General Chapter: Microbiological Attributes of Nonsterile Nutritional and Dietary Supplements, USP42-NF37, Rockville, MD, USA: United States Pharmacopeial Convention, 2019.

49. Food and Drug Administration [Internet]. Revised Guidelines for the Assessment of Microbiological Quality of Processed Foods. No. 2013-010; 2013 [cited 2021 Sep 22]. Available from: https://www.fda.gov.ph/wp-content/uploads/2021/03/FDA-Circular-No.-2013-010.pdf.>

50. Food and Drug Administration [Internet]. Bacteriological Analytical Manual Chapter 23: Methods for Cosmetics; 2021 [cited 2023 Feb 11]. Available from: https://www.fda.gov/food/laboratory-methods-food/bam-chapter-23-methods-cosmetics.

51. ISO 17516:2014. Cosmetics-Microbiology-Microbiological Limits. Geneva, Switzerland: International Organization for Standardization (ISO), 2014.

52. Foglia C, Allesina S, Amoruso A, De Prisco A, Pane M. New insights in enumeration methodologies of probiotic cells in finished products. J Microbiol Methods. 2020;175:105993.

53. Pasulka AL, Howes AL, Kallet JG, VanderKelen J, Villars C. Visualization of probiotics via epifluorescence microscopy and fluorescence in situ hybridization (FISH). J Microbiol Methods. 2021;182:106151.

54. Davis C. Enumeration of probiotic strains: Review of culture-dependent and alternative techniques to quantify viable bacteria. J Microbiol Methods. 2014;103:9-17.

55. United States Pharmacopeia, General Chapter: Microbiological Procedures for Absence of Specified Microorganisms – Nutritional and dietary supplements, USP42-NF37, Rockville, MD, USA: United States Pharmacopeial Convention, 2019.

56. European Pharmacopoeia, 10th ed., Strasbourg, France: EDQM, 2019; 6297-6301.

57. Russian Pharmacopoeia [Internet]. 13th ed. [cited 2021 Sep 17]. Available from: https://pharmacopoeia.ru/en/gosudarstvennaya-farmakopeya-xiii-online-gf-13-online/.

58. FAO/WHO. Working Group on Guidelines for the Evaluation of Probiotics in Food, 2002.

59. Best Practices Guidelines for Probiotics. The Council for Responsible Nutrition. The International Probiotics Association (IPA), 2017.

60. Doron S, Snydman DR. Risk and safety of probiotics. Clin Infect Dis. 2015;60(Suppl 2):S129-34.

61. Ben Braïek O, Smaoui S. Enterococci: Between emerging pathogens and potential probiotics. Biomed Res Int. 2019;2019:5938210.

62. Krawczyk B, Wityk P, Gałęcka M, Michalik M. The many faces of Enterococcus spp. – commensal, probiotic and opportunistic pathogen. Microorganisms. 2021;9(9):1900.

63. Hanchi H, Mottawea W, Sebei K, Hammami R. The genus Enterococcus: Between probiotic potential and safety concerns – An update. Front Microbiol. 2018;9:1791.

64. Huang M-CJ, Tang J. Probiotics in personal care products. Microbiol Discov. 2015;3:5.

65. França K. Topical probiotics in dermatological therapy and skincare: A concise review. Dermatol Ther (Heidelb). 2021;11(1):71-7.

66. Khalfallah G, Gartzen R, Möller M, Heine E, Lütticken R. A new approach to harness probiotics against common bacterial skin pathogens: Towards living antimicrobials. Probiotics Antimicrob Proteins. 2021;13(6):1557-71.

67. Zawistowska-Rojek A, Zareba T, Mrówka A, Tyski S. Assessment of the microbiological status of probiotic products. Pol J Microbiol. 2016;65(1):97-104.

68. Mohar Lorbeg P, Golob M, Kramer M, Treven P, Bogovič Matijašić B. Evaluation of dietary supplements containing viable bacteria by cultivation/MALDI-TOF mass spectrometry and PCR identification. Front Microbiol. 2021;12:700138.

69. Mazzantini D, Calvigioni M, Celandroni F, Lupetti A, Ghelardi E. Spotlight on the compositional quality of probiotic formulations marketed worldwide. Front Microbiol. 2021;12:693973.