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Food Standards Agency, & Food Standards Scotland. (2024). Safety Assessment on the safety and efficacy of Pediococcus ⁠pentosaceus DSM 32292 as a feed additive for all animal species (RP552). FSA Research and Evidence. https:/​/​doi.org/​10.46756/​001c.121317

Abstract

An application was submitted to the Food Standards Agency in March 2021 from Microferm Limited (‘the applicant’) for the new authorisation of an additive (Pediococcus pentosaceus DSM 32292), under the category of ‘technical additive’ and functional group ‘silage additives’. The additive is proposed to be used in all animal species, with a proposed minimum content of 1 x 1010 CFU / g to be applied to silage at a minimum of 5 x 107 CFU / kg.

To support the Food Standards Agency (FSA) and Food Standards Scotland (FSS) in evaluating the dossier, the Animal Feed and Feed Additives Joint Expert Group (AFFAJEG) and the Advisory Committee on Animal Feedingstuffs (ACAF) were asked to review the dossier and the supplementary information from the applicant.

The FSA/FSS concluded, based on the ACAF’s advice, that the additive was correctly identified and characterised, and that the additive can be considered safe for the target species, consumer, and the environment. The additive was identified as potentially harmful by contact with skin or eyes and a respiratory sensitiser.

It was concluded that the additive can be considered efficacious for moderately difficult to ensile forages. No conclusion could be reached on efficacy for difficult to ensile forages.

The views of AFFAJEG and ACAF have been taken into account in the safety assessment which represents the opinion of the FSA and FSS.

1. Introduction

The FSA and FSS have undertaken a safety assessment for a feed additive (Pediococcus pentosaceus DSM 32292 – Microferm Ltd., Spring Lane North, Malvern Link, Worcestershire, WR14 1BU, UK) under Assimilated Regulation (EC) No 1831/2003 (EC, 2003) under the category of ‘technical additives’ and functional group ‘silage additives’. The additive is proposed to be used in all animal species, with a proposed minimum content of 1 x 1010 CFU / g to be applied to silage at a minimum of 5 x 107 CFU / kg.

To support the safety assessment by FSA and FSS, the AFFAJEG and the ACAF provided advice to the FSA and FSS outlined in this document.

In line with Article 8 of 1831/2003, the assessment has considered whether the feed additive complies with the conditions laid down in Article 5, including: safety considerations for human, animal and environmental health; efficacy of the additive for its intended effect; potential impairment of the distinctive features of animal products. This, and the guidance put in place by EFSA for the evaluation of feed additive applications, has formed the basis and structure for the assessment.

The dossier was evaluated by the AFFAJEG at their May 2022 meeting and by the ACAF at their June 2023 meeting. Further information was provided by the applicant responding to queries by the FSA in June 2022, March 2023 and July 2023. The ACAF accepted the conclusions on this application at their April 2024 meeting.

The views of ACAF have been taken into account in this safety assessment which represents the opinion of the FSA/FSS.

Table 1.Table showing products included in this assessment
Title Product type Intended use/s Intended species or categories of animals
Pediococcus pentosaceus DSM 32292 Feed additive Silage additive All animal species

2. Assessment

2.1. Section II: Identity, characterisation and conditions of use

The additive is formulated to contain viable cells of Pediococcus pentosaceus DSM 32292, sucrose (5%), skimmed milk powder (20%), and glycerine (3%). The conditions of use proposed by the applicant are listed in Table 2. The additive specification table information is located within Table 3.

Table 2.Identity table for the additive Pediococcus pentosaceus DSM 32292
Composition
Pediococcus pentosaceus DSM 32292 Minimum 1x1010 cfu/g
Physico-chemical properties
Dusting potential 10.19 g/m3
Particle size distribution
(50 % of particles are less than)
134 µm
Purity
Microbial Impurities
Presumptive Coliforms < 10 CFU / g
Yeast < 100 CFU / g
Salmonella spp. Not Detected in 25 g
Presumptive E. coli < 10 CFU / g
Moulds < 100 CFU/g
Mycotoxins
Aflatoxin B1 ≤ 0.04 µg/kg
Aflatoxin B2 < 0.01 µg/kg
Aflatoxin G1 < 0.01 µg/kg
Aflatoxin G2 < 0.03 µg/kg
Total Aflatoxin ≤ 0.06 µg/kg
Heavy Metals
Arsenic ≤ 0.11 mg/kg
Lead ≤ 0.27 mg/kg
Cadmium ≤ 0.2 mg/kg
Mercury ≤ 0.02 mg/kg
Table 3.Proposed conditions of use of Pediococcus pentosaceus DSM 32292
Proposed mode of use in animal nutrition
Additive Pediococcus pentosaceus DSM 32292
Registration Number 1k1018
Category of additive Technical Feed Additive
Functional group of additive Silage Additive
Conditions of use
Species or category of animal Min. content Withdrawal Period
All animal species Min. 1 x 1010 CFU/g to be applied to silage at a Min. 5 x 10 7 CFU/kg.
  1. --

In the first evaluation, the AFFAJEG identified several pieces of information that the applicant would need to provide to inform the assessment of the product, including microbial resistance gene information, laboratory accreditation certificates, an additive label, and physical properties data for the additive. These queries were satisfactorily addressed by the applicant. The additive is a significantly dusty product, and measures should be taken to limit exposure through inhalation.

2.1.1. Conclusions on Section II

The ACAF concluded the additive was correctly identified and characterised. The product is dusty. No further concerns were raised for Section II of this dossier.

2.2. Section III: Safety

The Committee noted that the active substance (Pediococcus pentosaceus DSM 32292) is a qualified presumption of safety (QPS) microorganism and as such no toxicological tests were required to evaluate the safety for the target species, consumers, or the environment.

The applicant noted that the product will be labelled as a respiratory sensitiser. The ACAF concluded the additive is likely to form a dust that could be inhaled by workers. As no studies of skin or eye irritancy or skin sensitisation were provided, the Committee regard the additive as potentially harmful by contact with skin or eyes.

2.2.1. Conclusions on safety

The AFFAJEG concluded that the additive can be considered safe for target species, consumers, and the environment. The additive is likely to form a dust that has the potential to be inhaled by workers. The additive should be considered a respiratory sensitiser and potentially harmful by contact with skin and eyes.

2.3. Section IV: Efficacy

The applicant provided studies on the effect that Pediococcus pentosaceus DSM 32292 had within three separate ensiling trials on ‘moderately difficult to ensile’ forages.

  • Study A consisted of approximately 15 % perennial ryegrass, 60 % lucerne, 20 % red clover and 5 % herbs (mainly dandelion).

  • Study B was composed of approximately 30 % grasses (mainly timothy, meadow fescue and perennial ryegrass), 60 % red clover, 5 % lucerne, and 5 % herbs (mainly dandelion).

  • Study C consisted of approximately 25 % grasses (mainly timothy, meadow fescue and perennial ryegrass), 55 % red clover, 10 % white clover, and 10 % herbs (mainly dandelion).

The results demonstrated that the application of the P. pentosaceus inoculant improved the silage quality parameters pH, lactate, ethanol and 2,3-butanediol significantly in all three trials (Appendix A, Tables 4-6).

In the evaluation of the additive’s efficacy, the ACAF noted that the additive was only tested against a single class of forage. The Committee was able to conclude the additive was efficacious in moderately difficult to ensile forages. The Committee requested further information from the applicant however they were unable to provide studies within other classes of forage. Due to a lack of evidence the ACAF was unable to conclude on the additive’s efficacy in difficult to ensile forages.

2.3.1. Conclusions on efficacy

The Committee concluded that the additive is efficacious for moderately difficult to ensile forages. Due to a lack of evidence the ACAF was unable to conclude on the additive’s efficacy in difficult to ensile forages.

3. Analytical methods evaluation

Conclusions on the analytical methods are presented here as an extract from the Evaluation Report of the European Union Reference Laboratory for Feed Additives on the Method(s) of the Analysis for Pediococcus pentosaceus DSM 32292 (EURL-FA, 2020).

"For the identification of Pediococcus pentosaceus DSM 32292, the EURL recommends for official control (i) Whole Genome Sequencing Analysis (WGS) or (ii) Pulsed-Field Gel Electrophoresis (PFGE). The EURL considers that both methodologies are fit for purpose for the bacterial identification of authorised additives at a strain level.

For the enumeration of Pediococcus DSM 32292 in the feed additive, the EURL recommends for official control the ring-trial validated spread plate method EN 15786.

Since the unambiguous determination of the content of DSM 32292 initially added to silage is not achievable by analysis, the EURL cannot evaluate nor recommend any method for official control for the determination of Pediococcus pentosaceus DSM 32292 in silage."

FSA/FSS accepts the EURL analytical method evaluation reports. FSA/FSS determined the analytical method as appropriate for official controls for this feed additive.

4. Conclusions

The Committee concluded the additive was correctly identified and characterised. The additive shows a high dusting potential.

The ACAF concluded that the additive can be considered safe for target species, consumers, and the environment. The additive should be considered a respiratory sensitiser and potentially harmful by contact with skin and eyes.

The Committee concluded that the additive is efficacious for moderately difficult to ensile forages. Due to a lack of evidence the ACAF was unable to conclude on the additive’s efficacy in difficult to ensile forages.

The FSA/FSS agree with the conclusions reached by the AFFAJEG/ACAF. FSA/FSS accepts the EURL analytical method evaluation reports. FSA/FSS determined the analytical method as appropriate for official controls for this feed additive.


Acknowledgements

With thanks to the members of the AFFAJEG and ACAF during the course of the assessment, who were: Professor Nicholas Jonsson, Professor Emily Burton, Martin Briggs, Professor Katrina Campbell, Professor Matthew Fisher, Hannah Kane, Susan MacDonald, Dr. Oonagh Markey, Christine McAlinden, Dr. Donald Morrison, Derek Renshaw, Dr. Michael Salter, Dr. Adam Smith, Christel Wake, Dr. Helen Warren and Dr. Nick Wheelhouse.

Abbreviations

Acronym Definition
ACAF    Advisory Committee on Animal Feedingstuffs   
AFFAJEG    Animal Feed and Feed Additives Joint Expert Group   
CFU Colony Forming Units
DM Dry Matter
EC    European Commission   
EFSA    European Food Safety Authority   
EU    European Union    
EURL    European Union Reference Laboratory   
FSA    Food Standards Agency   
FSS    Food Standards Scotland   
FM Fresh Matter
GLP Good Laboratory Practices
N Total nitrogen
NH3-N Nitrogen content of ammonia
PFGE Pulsed-Field Gel Electrophoresis
Pr>Chi2 Probability > chi square
QPS Qualified Presumption of Safety
WGS Whole Genome Sequence
WSC Water-soluble carbohydrates

References

EC (European Commission). (2003). Regulation No 1831/2003 of the European Parliament and of the Council on additives for use in animal nutrition. https:/​/​www.legislation.gov.uk/​eur/​2003/​1831/​contents
EURL-FA (European Reference Laboratory for Feed Additives). (2020). Evaluation Report on the Analytical Methods submitted in connection with the Application for Authorisation of a Feed Additive according to Regulation (EC) No 1831/2003. Pediococcus DSM 32292. https:/​/​joint-research-centre.ec.europa.eu/​reports-and-technical-documentation/​fad-2021-0074_en

Appendix A: Efficacy tables

Table 4.Forage A (37% DM; 15 g WSC/kg FM) - Fermentation quality of P. pentosaceus-treated and untreated silages after 90 days.
DM g/kg FM WSC g/kg
DM
NH3-N g/kg N pH Lactic acid g/kg DM Acetic acid g/kg DM n-Butyric acid g/kg DM Propionic
acid g/kg DM
Succinic acid g/kg DM Ethanol g/kg DM 2,3-Butanediol g/kg DM
Control 370 8 80 5.57 20 11 1 3 8 8 5
Treated 370 1 70 4.50 76 8 <1 1 3 5 1
Pr>Chi2 0.77* 0.02 0.02 0.02 0.02 0.02 0.11* 0.013 0.02 0.02 0.02

Values represent treatment means from 4 replicate silos. DM: Dry matter; FM: Fresh matter; WSC: water-soluble carbohydrates; NH3-N: Nitrogen content of ammonia; N: Total nitrogen; Pr>Chi2: Probability>chi square; * notes no statistical significance.

Table 5.Forage B (36% DM; 17 g WSC/kg FM) - Fermentation quality of P. pentosaceus-treated and untreated silages after 90 days.
DM
g/kg
FM
WSC g/kg
DM
NH3-N g/kg N pH Lactic acid g/kg DM Acetic acid g/kg DM n-Butyric acid g/kg DM Propionic
acid g/kg DM
Succinic acid g/kg DM Ethanol g/kg DM 2,3- Butanediol g/kg DM
Control 359 14 67 5.46 16 10 <1 4 7 8 8
Treated 366 5 53 4.28 61 6 <1 1 2 5 1
Pr>Chi2 0.02 0.02 0.02 0.02 0.02 0.02 1.00 0.014 0.02 0.02 0.02

Values represent treatment means from 4 replicate silos. M: Dry matter; FM: Fresh matter; WSC: water-soluble carbohydrates; NH3-N: Nitrogen content of ammonia; N: Total nitrogen; Pr>Chi2: Probability>chi square; * notes no statistical significance.

Table 6.Forage C (34% DM; 24 g WSC/kg FM) - Fermentation quality of P. pentosaceus-treated and untreated silages after 90 days.
DM g/kg
FM
WSC g/kg
DM
NH3-N g/kg N pH Lactic acid g/kg DM Acetic acid g/kg DM n-Butyric acid g/kg DM Propionic
acid g/kg DM
Succinic acid g/kg DM Ethanol g/kg DM 2,3-Butanediol g/kg DM
Control 336 6 124 5.36 31 11 2.3 16 21 14 36
Treated 341 10 75 4.33 73 10 <1 7 10 6 8
Pr>Chi2 0.03 0.02 0.02 0.02 0.02 0.08 0.02 0.02 0.02 0.02 0.02

Values represent treatment means from 4 replicate silos. M: Dry matter; FM: Fresh matter; WSC: water-soluble carbohydrates; NH3-N: Nitrogen content of ammonia; N: Total nitrogen; Pr>Chi2: Probability>chi square; * notes no statistical significance.