Description
Pediococcus pentosaceus CECT 8330 is a gram-positive, non-spore-forming, live probiotic bacterium that was isolated from the human microbiome. It has been deposited in the Spanish Coleccion Espanola de Cultivos Tipo (CECT) under accession number, CECT 8330. The ingredient is manufactured under anaerobic fermentation conditions, concentrated, and freeze dried in the presence of cryoprotectants. Non-GMO corn starch-derived maltodextrin is then added for ingredient stability. The concentration of the final ingredient is >4 x 1011 colony-forming units (CFU)/g.
| Test | Method reference | Acceptance criteria |
|---|---|---|
| Characterisation | ||
| General properties | ||
| Colour and appearance | Visual | Ivory-white, free flowing powder |
| Solubility | USP <1236> | Slightly soluble in water, insoluble in methanol |
| Particle size | Pass through 1 mm sieve | 100% (w/w) |
| Loss on drying | USP <731> | <5% |
| Sulfated ash | Ph Eur 2.4.14 | <15% |
| pH | USP <791> | 6.5 - 7.5 |
| Water activity | Ph Eur 2.9.39 | <0.20 |
| Colony Morphology on MRS[1] medium (5% CO2) atmosphere | Visual | White, circular raised elevation and entire margin colonies |
| Microscopic Morphology on MRS[1] medium (5% CO2) atmosphere | Microscopic, Visual | Cocci (~0.6 - 1.0 mm); Tetrads form via cell division in two perpendicular directions in a single plane; distinctive of Pediococci |
| Gram staining | USP <1113> | Gram-positive |
| Antimicrobial resistance | EFSA[2] | Limited, non-transferable antimicrobial resistance to Kanamycin, Streptomycin and Tetracycline |
Antimicrobial susceptibility
| EFSA[2] | Susceptible to Ampicillin, Gentamicin, Erythromycin, Clindamycin and Chloramphenicol |
| Virulence factors | EFSA[3] | Absent |
| Pathogenic attributes | EFSA[3] | Probabilities of being a human pathogen of 0.172 and 0.223 (where a pathogen is 1.0) |
| Toxigenic attributes | OECD 420[4] | Absent |
| Identity | ||
| 16S rRNA Sequencing | USP <1113>
| Matches Pediococcus pentosaceus CECT 8330 |
| Whole-Genome fingerprint (PGFE) | USP <1113> | Matches Pediococcus pentosaceus CECT 8330 |
| Assay | ||
| Total Viable cell count | Ph Eur 5.1.6/EN 15786:2021[5] | >4 x 1011 CFU/g |
| Impurities and incidental constituents | ||
| Elemental impurities | ||
| Lead | USP <233> | Not more than 2.0 ppm |
| Arsenic | USP <233> | Not more than 1.5 ppm |
| Cadmium | USP <233> | Not more than 0.5 ppm |
| Mercury | USP <233> | Not more than 0.1 ppm |
| Microbial contaminants | ||
| Total aerobic microbial count (TAMC) | Ph Eur 2.6.13/Ph Eur 2.6.36 | <102 CFU/g |
| E. coli | Ph Eur 2.6.13/Ph Eur 2.6.36 | Not detected in 1 g |
| Bile-tolerant gram-negative bacteria | Ph Eur 2.6.13/Ph Eur 2.6.36 | Not detected in 1 g |
| S. aureus | Ph Eur 2.6.13/Ph Eur 2.6.36 | Not detected in 1 g |
| Salmonella | Ph Eur 2.6.13/Ph Eur 2.636 | Not detected in 1 g |
Footnotes
MRS (deMan, Rogosa, and Sharpe media) is an enriched selective medium for the cultivation of Lactobacillus. This animal-derived medium is not used in the production of Pediococcus pentosaceus CECT 8330.
EFSA (2018). Guidance on the characterisation of microorganisms used as feed additives or as production organisms, EFSA Journal,16(3), pp. 5206.
EFSA (2021). EFSA statement on the requirements for whole genome sequence analysis of microorganisms intentionally used in the food chain, EFSA Journal,19(7), pp. 6506.
OECD (2001). OECD Guideline for testing of chemicals: Acute oral toxicity - Fixed dose procedure [Online]. Available: https://ntp.niehs.nih.gov/sites/default/files/iccvam/suppdocs/feddocs/oecd/oecd_gl420.pdf [Accessed 28 April 2025].
EN 15786:2021: Animal feeding stuffs: Methods of sampling and analysis - Detection and enumeration of Pediococcus spp. used as feed additive.
Abbreviations
- BP: British Pharmacopoeia
- EFSA: European Food Safety Authority
- EMA: European Medicines Agency
- EN: Europäische Norm (European Standard)
- GMO: Genetically Modified Organism
- MRS: deMan, Rogosa, and Sharpe media
- OECD: Organisation for Economic Co-operation and Development
- PGFE: Pulsed-Field Gel Electrophoresis
- Ph Eur: European Pharmacopoeia
- USP: United States Pharmacopoeia