Peer review of the pesticide risk assessment of the active substance Trichoderma afroharzianum (formerly harzianum) T‐22

Abstract

The conclusions of the European Food Safety Authority (EFSA) following the peer review of the initial risk assessments carried out by the competent authorities of the rapporteur Member State, Sweden, and co‐rapporteur Member State, Netherlands, for the pesticide active substance Trichoderma afroharzianum (formerly harzianum) T‐22 are reported. The context of the peer review was that required by Commission Implementing Regulation (EU) No 844/2012. The conclusions were reached on the basis of the evaluation of the representative uses of Trichoderma afroharzianum (formerly harzianum) T‐22 as a fungicide on grass seed crops, root and tuber vegetables and stem vegetables. The reliable end points, appropriate for use in regulatory risk assessment, are presented. Issues that could not be finalised are identified.

SUMMARY

Commission Implementing Regulation (EU) No 844/2012 lays down the procedure for the renewal of the approval of active substances submitted under Article 14 of Regulation (EC) No 1107/2009. The list of those substances is established in Commission Implementing Regulation (EU) No 686/2012 as amended by Commission Implementing Regulation (EU) No 2016/183. Trichoderma afroharzianum (formerly harzianum) T‐22 is one of the active substances listed in that regulation.

In accordance with Article 1 of Regulation (EU) No 844/2012, the rapporteur Member State (RMS), Sweden, and co‐rapporteur Member State (co‐RMS), Netherlands, received an application from Koppert B.V. for the renewal of approval of the active substance Trichoderma afroharzianum strain T‐22.

An initial evaluation of the dossier on Trichoderma afroharzianum T‐22 was provided by the RMS in the renewal assessment report (RAR), and subsequently, a peer review of the pesticide risk assessment on the RMS evaluation was conducted by EFSA in accordance with Article 13 of Commission Implementing Regulation (EU) No 844/2012. The following conclusions are derived.

The representative uses of Trichoderma afroharzianum T‐22, applied as a fungicide by spraying on field crops of grass seed, root, tuber and stem vegetables, as well as in greenhouses and indoors for stem vegetables, as proposed at EU level, result in sufficient fungicidal efficacy against the target pests.

The assessment of the data package revealed no issues that could not be finalised or that need to be included as critical areas of concern with respect to the identity of the microbial active substance, physical and technical properties of the representative formulation and analytical methods. An issue not finalised was identified in relation to having insufficient evidence to exclude that Trichoderma afroharzianum T‐22 might have the biological property of being pathogenic to cereals including maize.

In the area of mammalian toxicology, the available evidence was not considered sufficient to exclude the presence of metabolites of concern in the MPCP. A data gap was identified for one metabolite and the risk from non‐dietary exposure could not be finalised.

In the area of residues, without data addressing the occurrence of emodin and pachybasin (non‐viable residues) including its potential in situ formation by Trichoderma afroharzianum T‐22 (viable residues) following the representative use conditions, the consumer risk assessment cannot be finalised.

None of the five criteria outlined in the guidance (SANCO/11188/2013 Rev. 2) were considered met for Trichoderma afroharzianum T‐22.

With respect to fate and behaviour in the environment, Trichoderma afroharzianum T‐22 can be expected to persist and multiply in soil in concentrations considerably higher than the natural background levels, taking into account repeated applications over the years. Currently, the available information is insufficient to determine whether the metabolites of potential concern (e.g. emodin) meet the criteria set out in Part B, Section 7 (iv) of Commission Regulation (EU) No 283/2013 and on their presence in the MPCP and/or their in situ production in the environment. A data gap was identified for the exposure of secondary metabolite of potential concern for environmental compartments.

In the area of ecotoxicology, it is not possible to conclude on the infectivity and pathogenicity of Trichoderma afroharzianum T‐22 for non‐target arthropods other than bees. In addition, the risk assessment to non‐target organisms exposed to secondary metabolites of potential concern could not be finalised.

BACKGROUND

Commission Implementing Regulation (EU) No 844/2012 ¹ (hereinafter referred to as ‘the Regulation’) lays down the provisions for the procedure of the renewal of the approval of active substances submitted under Article 14 of Regulation (EC) No 1107/2009. ² This regulates for the European Food Safety Authority (EFSA) the procedure for organising the consultation of Member States, the applicant(s) and the public on the initial evaluation provided by the rapporteur Member State (RMS) and/or co‐rapporteur Member State (co‐RMS) in the renewal assessment report (RAR), and the organisation of an expert consultation where appropriate.

In accordance with Article 13 of the Regulation, unless formally informed by the European Commission that a conclusion is not necessary, EFSA is required to adopt a conclusion on whether the active substance can be expected to meet the approval criteria provided for in Article 4 of Regulation (EC) No 1107/2009 within 5 months from the end of the period provided for the submission of written comments, subject to an extension of an additional 3 months where additional information is required to be submitted by the applicant(s) in accordance with Article 13(3).

In accordance with Article 1 of the Regulation, the RMS, Sweden, and co‐RMS, Netherlands, received an application from Koppert B.V. for the renewal of approval of the active substance Trichoderma afroharzianum (formerly harzianum) T‐22. Complying with Article 8 of the Regulation, the RMS checked the completeness of the dossier and informed the applicant, the co‐RMS (The Netherlands), the European Commission and EFSA about the admissibility.

The RMS provided its initial evaluation of the dossier on Trichoderma afroharzianum T‐22 in the RAR, which was received by EFSA on 31 March 2021 (Sweden, 2021).

In accordance with Article 12 of the Regulation, EFSA distributed the RAR to the Member States and the applicant, Koppert B.V., for consultation and comments on 21 July 2021. EFSA also provided comments. In addition, EFSA conducted a public consultation on the RAR. EFSA collated and forwarded all comments received to the European Commission on 21 September 2021. At the same time, the collated comments were forwarded to the RMS for compilation and evaluation in the format of a reporting table. The applicant was invited to respond to the comments in column 3 of the reporting table. The comments and the applicant's response were evaluated by the RMS in column 3.

The need for expert consultation and the necessity for additional information to be submitted by the applicant in accordance with Article 13(3) of the Regulation were considered in a teleconference between EFSA and the RMS on 17 February 2022. On the basis of the comments received, the applicant's response to the comments and the RMS's evaluation thereof, it was concluded that additional information should be requested from the applicant, and that EFSA should conduct an expert consultation in the areas of potential effects on human/mammalian health and potential effects on non‐target organisms.

The outcome of the teleconference, together with EFSA's further consideration of the comments, is reflected in the conclusions set out in column 4 of the reporting table. All points that were identified as unresolved at the end of the comment evaluation phase and which required further consideration, including those issues to be considered in an expert consultation, were compiled by EFSA in the format of an evaluation table.

The conclusions arising from the consideration by EFSA, and as appropriate by the RMS, of the points identified in the evaluation table, together with the outcome of the expert consultation and the written consultation on the assessment of additional information, where these took place, were reported in the final column of the evaluation table.

A final consultation on the conclusions arising from the peer review of the risk assessment took place with Member States via a written procedure in June–July 2026.

This conclusion report summarises the outcome of the peer review of the risk assessment of the active substance and the formulation for representative uses, evaluated on the basis of the representative uses of Trichoderma afroharzianum T‐22 as a fungicide on grass seed crops, root and tuber vegetables, and stem vegetables, as proposed by the applicant. In accordance with Article 12(2) of Regulation (EC) No 1107/2009, risk mitigation options identified in the RAR and considered during the peer review, if any, are presented in the conclusion.

A list of the relevant end points for the active substance and the formulation is provided in Appendix A.

A key supporting document to this conclusion is the peer review report (EFSA, 2026), which is a compilation of the documentation developed to evaluate and address all issues raised in the peer review, from the initial commenting phase to the conclusion. The peer review report comprises the following documents, in which all views expressed during the course of the peer review, including minority views, where applicable, can be found:

• the comments received on the RAR;

• the reporting table (23 August 2021);

• the evaluation table (31 July 2026);

• the reports of the scientific consultation with Member State experts (where relevant);

• the comments received on the assessment of the additional information (where relevant);

• the comments received on the draft EFSA conclusion.

Given the importance of the RAR, including its revisions (Sweden, 2025), and the peer review report, both documents are considered as background documents to this conclusion and thus are made publicly available.

It is recommended that this conclusion and its background documents would not be accepted to support any registration outside the EU for which the applicant has not demonstrated that it has regulatory access to the information on which this conclusion report is based.

THE MICROBIAL ACTIVE SUBSTANCE AND THE FORMULATION FOR REPRESENTATIVE USES

The active substance Trichoderma afroharzianum T‐22 has been deposited in the American Type Culture Collection with the accession number ATCC 20847. Trichoderma afroharzianum T‐22 is a laboratory induced mutant strain obtained in vitro as an UV‐treated protoplast fusion hybrid of the Trichoderma harzianum T95 and Trichoderma harzianum T12, both also deposited in the American Type Culture Collection with the accession numbers 60850™ and 20737™, respectively. The notifier did not provide all known differences between the modified microorganism and the parent wild strains (data gap, see Section 10). Trichoderma afroharzianum T‐22 was previously identified as Trichoderma harzianum; however, based on recent results of phylogenetic studies, the strain T‐22 was re‐classified as Trichoderma afroharzianum.

The microbial pest control product (MPCP) for the evaluation of the representative uses is ‘Trianum‐P’, a wettable granule (WG) containing 1 × 10¹² to 2.4 × 10¹² CFU/Kg (nominal content: 1 × 10¹² CFU/Kg) of Trichoderma afroharzianum T‐22.

The information on the active substance and the formulation for representative uses, including the co‐formulants in this formulation, was considered in the overall assessment during the peer review. None of the co‐formulants is an unacceptable co‐formulant listed in Annex III of Regulation (EC) No 1107/2009. ³ One co‐formulant is a currently approved active substance under Regulation (EC) 1107/2009, ⁴ however, as it is approved for a different function, is not expected to contribute to the efficacy of the product as a fungicide. ⁵ Details on the composition of the formulations cannot be reported because of the provisions in Article 63(2)(d) of Regulation (EC) No 1107/2009; however, this information was fully available and evaluated during the peer review. A proposal for classification of the formulation according to Regulation (EC) 1272/2008 was provided by the applicant and assessed by the RMS (please see Volume 3 MP of the RAR).

The representative EU uses evaluated were spray applications on grass seed, tuberous plants, root crops, stem vegetables in the field and spray applications on stem vegetables in greenhouses (permanent) and temporary protected structures (e.g. walk‐in tunnels) and indoors, as a fungicide against Pythium, Rhizoctonia, Fusarium, Clarireedia and Sclerotinia. Full details of the GAP can be found in the list of end points in Appendix A.

Data were submitted to conclude that the use of Trichoderma afroharzianum T‐22 according to the representative uses proposed at the EU level results in sufficient fungicidal efficacy following the guidance document SANCO/2012/11251‐rev. 4 (European Commission, 2014).

General aspects

With regard to mammalian toxicity, the toxicological profile of the formulation for representative uses was discussed in the Pesticides Peer Review Teleconference (TC) 159 ⁶ (see Section 3). Acute toxicity, pathogenicity and infectiveness and sensitisation studies were available for the formulation for representative uses ‘TRIANUM‐P’.

With regard to the co‐formulants contained in ‘TRIANUM‐P’, sufficient toxicological data were available to address the genotoxicity and repeated dose toxicity potential of ‘TRIANUM‐P’, over the short and long term and no concerns were identified. ⁷

The availability of ecotoxicity data with the formulation for representative uses was discussed at the Pesticides Peer Review Experts' TC 161 ⁸ (refer to Section 6). The experts also discussed the data retrieval search and the available data for the individual components and no concerns were identified.

1. SPECIFICATION OF THE MICROORGANISM, PHYSICAL AND TECHNICAL PROPERTIES AND METHODS OF ANALYSIS

The following guidance documents were followed in the production of this conclusion: European Commission (2000, 2012), EFSA FEEDAP Panel (2012).

The microbial pest control agent as manufactured (MPCA‐AM) is only a hypothetical stage in the continuous production process of the formulation for the representative uses. As a consequence, the specification is provided only for the formulation ‘Trianum‐P' based on batch data from industrial scale plant production. The minimum and maximum content of Trichoderma afroharzianum T‐22 in the ‘Trianum‐P’ was 1.0 × 10¹² – 2.4 × 10¹² CFU/kg, and the nominal was 1.0 × 10¹² CFU/kg. The content of microbial contaminants of the ‘Trianum‐P’ was below the limits defined in the Working Document on Microbial Contaminant Limits (European Commission, 2012).

Data for ‘Trianum‐P’ were not provided for the flammability ⁹ and storage stability, ¹⁰ including the content of active substance and microbial contaminants after storage (data gap, see Section 10).

Identification of the active substance at strain level is achieved through sequence analysis of the ITS1 and tef1 regions. Trichoderma afroharzianum T‐22 can be distinguished from its parent strain T‐95 by isoenzyme electrophoresis and from its parent strain T‐12 using 12 marker sequences derived from high‐quality genomic sites, validated by PCR and subsequent Sanger sequencing.

Acceptable methods were available for the determination of the microorganism and the content of contaminating microorganisms in ‘Trianum‐P’. Residue definitions were not set for Trichoderma afroharzianum T‐22 (see Sections 4 and 6); therefore, monitoring methods are not required.

2. BIOLOGICAL PROPERTIES OF THE MICROORGANISM

Historical background: Historically Trichoderma species have been used as microbial biological control agents in agriculture, but also for enzyme and other chemicals production in food industries, wine and beer industries, textile industries, medical applications and for bioremediation.

Natural occurrence and geographical distribution: Trichoderma afroharzianum T‐22 mainly occurs in the soil and it does not occur naturally as it is a laboratory induced mutant strain. Wild strains of Trichoderma afroharzianum have been identified in Africa and Asia, but not in Europe.

The target organisms for the representative uses are soil‐borne fungal pathogens like Pythium, Rhizoctonia, Fusarium, Clarireedia, Microdochium and Sclerotinia.

The provided information indicates that the mode of action of Trichoderma spp. involves competition for nutrients, antibiosis, mycoparasitism and induced resistance in plants.

Infectiveness, dispersal and colonisation ability: Non‐Good Laboratory Practice (non‐GLP) data indicated that when grown on a potato dextrose medium, Trichoderma afroharzianum T‐22 grows at temperatures between 10°C and 30°C, but not at 37°C.

Plant pathogenicity: Trichoderma afroharzianum has recently been reported as a plant pathogenic fungus on maize (Zea mays) in several regions of Europe, where it has been associated with ear rot disease. The first cases were described in 2018, and the species is currently included in the EPPO Alert List as a causal agent of ear rot on maize (EPPO RS 2022/087; 2023/194; 2024/140, 2025). In addition to the references cited in the EPPO reports, recent publications in peer‐reviewed journals and on preprint servers have provided further evidence on the pathogenicity of Trichoderma strains on maize (Harish et al., 2025; Pfordt et al., 2024; Pfordt, Douanla‐Meli, Schäfer, et al., 2025; Pfordt, Douanla‐Meli, Voll, et al., 2025; Pfordt & von Tiedemann, 2025). In this context, the possibility that T. afroharzianum T‐22 may exert harmful effects on maize or on other plant species cannot be excluded on the basis of the information currently available in the dossier assessed, which was submitted by the applicant before the plant pathogenic nature of T. afroharzianum had been fully recognised. Therefore, robust evidence is required to demonstrate that Trichoderma afroharzianum T‐22 does not cause adverse effects on maize (data gap, see Section 9.1 ). As this has been identified as an issue not finalised, an appropriate limitation of use, according to Commission Regulation (EU) No 546/2011 ¹¹ Annex Part II C 2.4.2, may need to be considered by risk managers to prevent unacceptable effects on treated plants and adjacent and succeeding crops.

Secondary metabolites of (potential) concern: Based on available literature, several metabolites of potential concern can be produced by Trichoderma afroharzianum T‐22, such as trichothecenes, diketopiperazine‐ like compounds (e.g. gliovirin and gliotoxin), peptaibols (e.g. harzianins, alamethicin), volatile organic compounds (including 6‐PP) polyketides (e.g. tricholignan B, T‐22‐azaphilone), anthraquinones (e.g. pachybasin, emodin) and alkaloids (e.g. camptothecin and cytochalasans).

The genetic capacity of producing peptaibols and polyketides has been confirmed by the whole genome sequencing (WGS), while evidence indicates that the strain lacks the genetic capacity to produce trichothecenes and diketopiperazine‐ like compounds (e.g. gliovirin and gliotoxin). The genetic capacity to produce the volatile organic compound (VOC) 6‐PP, anthraquinones (e.g. emodin and pachybasin) and alkaloids (e.g. cytochalasans and camptothecin) was not investigated.

A non‐GLP screening test on several MPCP batches detected the presence of 6‐PP, emodin, tricholignan B and several harzianins. Several other peptaibols (e.g. alamethicin) were screened and not detected. Analytical quantification of emodin, 6‐pentyl‐2H‐pyran‐2‐one (6‐PP), and alamethicin in a GLP study confirmed that these metabolites were not detected above the limit of detection (LOD) in the MPCP (additional details about analytical determination of the individual metabolites belonging to the families mentioned above are reported in Appendix A). Further considerations on the potential concern for human and animal health are provided in Section 3.

Antifungal susceptibility: Non‐GLP data on testing Trichoderma afroharzianum T‐22 with several antimycotic agents allowed to determine the minimum inhibitory concentration (MIC) for amphotericin B, voriconazole, caspofungin and ketoconazole. These antimycotic agents correspond to three different modes of action. For fluconazole, itraconazole and flucytosine, no MICs could be derived, as even at the maximum applied concentrations, growth inhibition could not be observed. It is noted that EUCAST breakpoints for Trichoderma afroharzianum species together with the tested antifungal agents were not reported to allow the strain to be clearly categorised as resistant or susceptible to tested substances. None of the antibiotics used in human and veterinarian medicine are known to be produced by the active microorganism.

3. POTENTIAL EFFECTS ON HUMAN/MAMMALIAN HEALTH

The toxicological profile of Trichoderma afroharzianum T‐22 and its metabolites was discussed at the Pesticides Peer Review TC 159 (31 January 2025).

3.1. Mammalian infectivity and pathogenicity of the microorganism and sensitisation effects

As regards medical information, there is no evidence that Trichoderma afroharzianum species causes infection in immuno‐competent healthy humans. No adverse health effects have been reported in workers as a result of exposure to Trichoderma afroharzianum T‐22 during production and handling.

Trichoderma afroharzianum is not included in the EFSA qualified presumption of safety (QPS) list, as filamentous fungi are currently excluded from QPS evaluation due to safety and complexity considerations (EFSA BIOHAZ Panel, 2025).

Current methods for testing dermal sensitisation are not suitable for testing microorganisms, and no validated test methods exist for assessing sensitisation via inhalation. Based on their characteristics, microorganisms such as Trichoderma afroharzianum T‐22 shall be regarded as potential sensitisers.

Due to their pathogenic effects on other fungi, several Trichoderma species are used in biocontrol. There is no known specific human or animal (pets or livestock) pathogen within the genus Trichoderma; however, some Trichoderma species can act as opportunistic human pathogens.

No signs of toxicity or pathogenicity have been detected (technical grade material) after acute administration by oral, intratracheal, intraperitoneal and intravenous route in rats of the active substance Trichoderma afroharzianum T‐22. Microbial analysis of faeces, fluids and tissues/organs was only carried out after oral administration of the formulation for representative uses ‘TRIANUM P’ and demonstrated full clearance (no infectivity) of the active organism.

Skin and eye irritation were not observed in rabbits after exposure to the formulation for representative uses, as well as skin sensitisation. Nevertheless, the use of personal protective equipment (PPE) and respiratory protective equipment (RPE) for operators and workers can be considered as non‐specific risk mitigation measure considering that microorganisms are also potential sensitisers.

It is concluded that Trichoderma afroharzianum T‐22 has no potential for infectivity/pathogenicity in humans based on the available data.

3.2. Mammalian toxicity of the metabolites of (potential) concern

Trichoderma afroharzianum T‐22 can produce peptaibols, certain polyketides, anthraquinones (e.g. emodin and pachybasin), volatile organic compounds (e.g. 6‐PP) and alkaloids (e.g. cytochalasans and camptothecin) (see Section 2).

For peptaibols, in vitro cytotoxicity has been reported in boar sperm and somatic cells. It is known that peptaibols cause low acute toxicity in vivo. Therefore, based on the available limited information, they are unlikely to be metabolites of potential concern for human health. Alamethicin, a peptaibol, was below the LOD in the MPCP.

Emodin (belonging to the anthraquinones’ family) is considered a potential genotoxic compound since it is structurally related to the known genotoxic aloe‐emodin (EFSA ANS Panel, 2018). Pachybasin, also belonging to anthraquinones in the polyketides' families, showed low toxicity potential in acute and repeated dose studies in mice. No evidence of genotoxicity has been reported in the available literature review. However, in view of the structural alert related to the presence of the anthraquinone ring system, its genotoxic potential should be further clarified.

Emodin was not investigated in the WGS analysis and resulted below the LOD in the MPCP (see Section 2). No WGS analysis nor analytical determination has been provided for pachybasin (data gap, see Section 9.1).

It is noted that 6‐PP (6‐pentyl‐2H‐pyran‐2‐one) is a volatile organic compound authorised as a food flavouring substance (FL no. 10.031) by Commission Implementing Regulation (EU) No 872/2012. ¹²

Based on the assessment as a flavouring substance, a NOAEL of 12.1 mg/kg bw per day was identified in a 90‐day study in rats with 6‐PP and it was concluded not to pose safety concern at estimated levels of intake (EFSA CEF Panel, 2011) (see Section 4). Available literature information indicates that it would not be of toxicological concern, with the exclusion of possible local effects in the respiratory system, such as irritation and inflammation. The analytical quantification demonstrated that 6‐PP levels in the MPCP are below the LOD (see Section 2). Adverse effects in the respiratory tract are not expected to be of concern for non‐dietary exposure, considering that personal protective equipment PPE are required for PPPs. For bystanders and residents, the non‐dietary exposure resulting from the supported uses is not a concern considering that a safe consumption level has been identified as a food flavouring and that the levels in the field (i.e. during/after application) are not expected to trigger an adverse effect by inhalation.

Based on the available literature data, information on adverse effects was not retrieved for polyketides, alkaloids (e.g. camptothecins and cytochalasins), anthraquinones (other than emodin) and the other metabolites identified in the screening (see Section 2).

The RMS and Member States (MSs) supported the view that no metabolites of concern for human health are identified associated with Trichoderma afroharzianum T‐22 under the proposed conditions of use of the representative formulation ‘Trianum‐P'. The RMS and MSs were also in agreement that the in situ production of metabolites is not considered of concern.⁶

Based on the available toxicological information and analytical results, EFSA identified no concern for non‐dietary exposure to the product, except in relation to pachybasin (data gap, Section 9.1), unless its concentration is demonstrated to be below the threshold for genotoxicity classification. Contrary to the view of the RMS and MSs, EFSA considers that if in situ production of secondary metabolites of potential toxicological concern (such as emodin and pachybasin) is demonstrated, this may trigger further toxicological investigation.

4. RESIDUES

The assessment in the residue section is based on the following guidance documents: Commission guidance SANCO/11188/2013 Rev. 2 (European Commission, 2015). Trichoderma afroharzianum T‐22 is included in Annex IV of Regulation (EC) No 396/2005.

With regard to the criteria outlined in the guidance for an assessment of active substances for a possible inclusion into Annex IV of Regulation (EC) No 396/2005, Trichoderma afroharzianum T‐22 does not meet criterion I and II (not approved as basic substance and not listed in Annex I of Regulation (EC) No 396/2005).

Trichoderma afroharzianum T‐22 may produce emodin and pachybasin, representing hydroxyanthraquinone/anthracene derivatives, as secondary metabolites of potential concern (see Section 3.2). Their occurrence on edible crops relevant for their potential in situ production as non‐viable residues, has not been investigated (data gap on non‐viable residues; see Section 9.1).

Based on the current information, peptaibols are unlikely to be of concern.

A data gap was set for qualitative and quantitative information on the residues identified as metabolites of potential toxicological concern (emodin and pachybasin) in edible commodities under conditions of the representative uses on stem vegetables and on root and tuber vegetables. Demonstration of the levels of these metabolites in edible commodities is needed. Therefore, criterion III of the above‐mentioned guidance that there are no hazardous properties related to Trichoderma afroharzianum T‐22 cannot be concluded.

The RMS is of the view that neither non‐viable residues of toxicological relevance nor viable residues of potential concern are expected to occur. This view is supported by one Member State (see also Section 3.2.).

The representative uses consist of soil spray applications at BBCH 00 on root and tuber vegetables, whereas stem vegetables may receive a second application up to harvest, making this the more critical use pattern.

Information on absence of viable residues of Trichoderma afroharzianum T‐22 at harvest on carrots following treatments matching the representative uses on root and tuber vegetables is available. However, viable residues were not investigated during crop development, and therefore, the potential in situ formation of non‐viable residues cannot be excluded for root and tuber vegetables. Data were also not available for the residue behaviour of viable residues which could potentially form toxicologically relevant metabolites in situ up to harvest for stem vegetables, representing the more critical use pattern due to the possibility of a second application (data gap on viable residues, see Section 9.1 ). Supporting information on dispersion of viable residues of Trichoderma atroviride SC1 following soil treatment reports significant higher residues in treated (100 times higher in bottom and 10 times higher in top leaves of grapevine plants) than in untreated plants. Therefore, dispersal of residues from soil onto edible plant parts cannot be excluded for Trichoderma afroharzianum T‐22.

Based on the available scientific data, it cannot be concluded that non‐viable residues belonging to the anthraquinone family (for the representative uses on stem and root and tuber vegetables) would occur at levels below a potential health concern for the consumer. There is also insufficient information to conclude whether exposure to non‐viable residues linked to the representative uses of Trichoderma afroharzianum T‐22 is lower, equal to or higher than concentrations naturally occurring (data gap on non‐viable residues, see Section 9.1 ) Therefore, it cannot be concluded whether criterion IV of the above‐mentioned guidance is met.

The potential production of metabolite 6‐PP does not raise a concern for consumers. This conclusion is supported by a previous assessment of 6‐PP as a food flavouring (see Section 3.2), in which no safety concerns were identified at an estimated intake of 84 μg/person per day. Should Trichoderma afroharzianum T‐22 produce 6‐PP, residues on crops are likely to be very low, if present at all. Consequently, dietary exposure is expected to be well below the intake level established as safe by the EFSA CEF Panel in its assessment of 6‐PP as a flavouring.

Consumer exposure to viable cells of Trichoderma afroharzianum T‐22 on stem vegetables and to its potentially in situ formed metabolites on stem and root, and tuber vegetables cannot be excluded for the representative uses. Therefore, criterion V of the above‐mentioned guidance concerning a possible exclusion of dietary consumer exposure to Trichoderma atroviride T‐22 is not considered to be met.

Consequently, the consumer risk assessment could not be finalised (see Section 9.1). With regard to the five assessment criteria according to the Commission guidance SANCO/11188/2013 Rev. 2 (European Commission, 2015), none of the criteria were considered met for Trichoderma afroharzianum T‐22.

5. ENVIRONMENTAL FATE AND BEHAVIOUR

Filamentous fungi such as Trichoderma afroharzianum T‐22, including their conidia, are not expected to interfere with the analytical systems for the control of the quality of drinking water provided for in Directive 98/83/EC ¹³ (see specific Annex VI decision‐making criteria in Part II of Commission Regulation (EU) No 546/2011 ¹⁴ ).

Trichoderma afroharzianum T‐22 is a laboratory induced mutant not existing previously in nature (see Section 2). Therefore, natural background levels of this strain in Europe should be considered inexistent before its introduction as a biopesticide. At species level, Trichoderma afroharzianum has been reported to exist naturally in Africa and Asia (see Section 2).

Trichoderma afroharzianum has been recently identified as a pathogenic fungus in maize in certain regions of Europe being attributed as the cause of ear‐rot disease, which has been described recently (2018) for the first time in Europe, for which an EPPO alert is currently active (see Section 2).

5.1. Fate and behaviour in the environment of the microorganism and any other forms

The persistence and multiplication in soil was reported in a peer‐reviewed published study on the application of Trichoderma afroharzianum T‐22 to onion and carrot seed during drum priming, and their subsequent survival and establishment in the rhizosphere once the seed was planted. Trichoderma afroharzianum T‐22 remained relatively constant in number throughout the 8‐week period of the experiment but showed better survival on carrot than onion roots. Similar results were found in three different soil types (light sandy loam, peat soil and sandy clay loam (mixed 4:1) with vermiculite to improve its structure).

Trichoderma species were monitored in agricultural soil in response to application of biopreparations to lettuce crops using multiplex‐PCR. Indigenous Trichoderma species were identified in field soil; however, their abundance was estimated to be relatively low 10 ³ CFU, when compared to 10⁵ CFU g⁻¹ of dry soil after biopreparations application, and these fungi persisted at this level even after 2 years.

The information provided in these peer reviewed scientific literature studies indicates that Trichoderma spp. may persist overwinter and that this is achieved through the formation of chlamydospores.

Overall, the information on the species was sufficient to conclude that Trichoderma afroharzianum T‐22 can be expected to persist for over 2 years and multiply in soil in concentrations considerably higher than the background levels for the species, taking into account repeated applications over the years (Commission Regulation (EU) No 546/2011). It is acknowledged that there are many abiotic and biotic factors influencing persistence, survival and growth of the microorganism and that there are natural fluctuations in the population densities. Predicted environmental densities (PED) in soil (see Appendix A) can be used for a preliminary risk assessment assuming single annual use of Trichoderma afroharzianum T‐22. However, for repeated applications across multiple years, further data may be required to determine the potential accumulation levels of Trichoderma afroharzianum T‐22 in soil over time. This requires particular attention due to the recently identified uncertainty associated with the pathogenicity of Trichoderma afroharzianum to maize crops (see Section 2). The applicant should demonstrate that the available PED estimates adequately address plateau levels following repeated seasonal applications and provide information to show whether populations decline over time after the last application (data gap, see Section 10 ).

Specific studies on the mobility in soil of Trichoderma afroharzianum T‐22 were not available in the dossier. However, literature data presented in the RAR indicate that Trichoderma species can migrate within a few metres horizontally and a few decimetres vertically from the application site within few weeks of last application. Using the results from these studies, the horizontal and vertical mobility of Trichoderma afroharzianum T‐22 in soil is expected to be limited, although some dispersal to adjacent soil habitats cannot be excluded under the representative field and temporary protection production system (e.g. walk‐in tunnel) uses. Given the recently identified concerns regarding the pathogenicity of Trichoderma afroharzianum to maize crops, exposure of non‐target plants in the vicinity of the treated crops might require investigation.

No specific studies on the persistence and multiplication in surface water and sediment of Trichoderma afroharzianum T‐22 were available. Trichoderma afroharzianum T‐22 has been assessed as not pathogenic to aquatic organisms (see Section 6). Predicted environmental densities (PED) in surface water were provided based on the loading through spray drift.

Inadequate information was provided on the survival, competitiveness and population dynamics of Trichoderma afroharzianum T‐22 in air. However, conidia and spores of Trichoderma afroharzianum T‐22 may be dispersed by air to nearby areas surrounding the application site. Exposure to the air compartment and to other compartments through air cannot be excluded from the representative uses as spray application.

5.2. Fate and behaviour in the environment of secondary metabolite(s) of (potential) concern produced by the microorganism under relevant conditions

Several metabolites identified as being of potential concern (see Section 2) were screened in 11 MPCP batches, and selected compounds were quantified in five additional batches. The low levels detected, or the absence of detection, for the screened metabolites indicate that no more than approximately 1 g/ha of these secondary metabolites would be applied under the proposed uses of the MPCP. In most cases, such levels can be considered environmentally insignificant, provided that no severe toxicological or ecotoxicological concerns are associated with the metabolites in question.

Nevertheless, not all metabolites reported in the literature were included in the screening, and only a subset was quantified in the MPCP. Furthermore, no information is available on the potential in situ production of secondary metabolites by T. afroharzianum T‐22 following application. Therefore, a data gap is identified for information on the production of secondary metabolites considering their levels present in the environment after product application and their potential toxicity (e.g. genotoxic emodin and pachybasin, see Sections 2, 3.2, 6 and 9.1) in order to conclude on the exposure of these metabolites to the environment (soil, surface water, groundwater) according to the representative uses except for the uses indoors and in greenhouses (permanent), in relation to groundwater.

6. POTENTIAL EFFECTS ON NON‐TARGET ORGANISMS

The ecotoxicology risk assessment was discussed in the Pesticides Peer Review Experts' TC 161 (February 2025).

6.1. Birds and mammals

Studies for birds are available with the active substance and the formulation for representative uses. In both studies, no adverse effects were reported. The available data do not indicate that the active substance is infective or pathogenic to birds.

For mammals, acute toxicity studies are available with the active substance. The available data do not indicate that the active substance is infective or pathogenic to wild mammals.

The available literature studies and whole‐genome sequencing analyses revealed the potential for Trichoderma afroharzianum T‐22 to produce various secondary metabolites of potential concern (see Section 2). The hazard characterisation and exposure assessment of secondary metabolites produced by Trichoderma afroharzianum T‐22 could not be finalised with the limited data set (see Section 5.2). For birds and wild mammals, a low risk cannot be concluded for the intended uses in the field and temporary protected cropping systems (e.g. walk‐in tunnels) (data gap leading to an issue that could not be finalised, see Section 9.1). Based on the representative use in greenhouses (permanent) and in line with EFSA (2015), minimal exposure to secondary metabolites is expected; hence, low risk can be concluded for birds and mammals for the uses indoors and in greenhouses (permanent). Nevertheless, it is noted that the RMS disagreed and considered that, for all representative uses, secondary metabolites are expected to be of limited or no concern and a low risk can be concluded.

6.2. Aquatic organisms

One study for fish is available with the active substance. Studies with the representative formulation are available for aquatic invertebrates. Furthermore, published literature studies with Trichoderma asperellum ICC012 were provided for fish, aquatic invertebrates, algae and aquatic plants. These studies were accepted to be used in the assessment for Trichoderma afroharzianum T‐22.

The available data do not indicate that Trichoderma afroharzianum T‐22 is infective or pathogenic to any aquatic organisms.

No indication is available with regard to the production of secondary metabolites in the aquatic environments. Toxicity data were not available for secondary metabolites to perform a hazard characterisation (data gap leading to an issue that could not be finalised, see Section 9.1). The risk assessment for secondary metabolites could not be finalised for aquatic organisms (fish, freshwater invertebrates, algae and aquatic plants other than algae) for the intended uses (see also Section 5.2). The RMS disagreed and considered that, for all representative uses, secondary metabolites are expected to be of limited or no concern and a low risk can be concluded.

6.3. Bees

One study is available for honey bees with the representative formulation. Two published studies with the representative formulation are available for bumble bees. Furthermore, literature studies for honey bees and bumble bees were provided with other Trichoderma spp., and one published 3‐year study on honey bees and bumblebees with T. afroharzianum T‐22. These studies were accepted to be used in the assessment for Trichoderma afroharzianum T‐22.

Based on the results of the studies, no clear or consistent indication of infectivity and pathogenicity is concluded. However, no information was available demonstrating that the potentially produced metabolites of concern were present in the studies. Toxicity data were not available for secondary metabolites to perform a hazard characterisation (data gap leading to an issue that could not be finalised, see Section 9.1). The risk assessment for secondary metabolites could not be finalised for honey bees for the representative uses in field and temporary protected cropping systems (e.g. walk‐in tunnels) (see also Section 5.2). Based on the representative use indoors and in greenhouses (permanent) and in line with EFSA (2015), minimal exposure to secondary metabolites is expected; hence, low risk can be concluded for bees for the uses indoors and in greenhouses (permanent). The RMS disagreed and considered that, for all representative uses, secondary metabolites are expected to be of limited or no concern and a low risk can be concluded.

6.4. Non‐target arthropods other than bees

No studies were available with Trichoderma afroharzianum T‐22 or the representative formulation. Studies with other Trichoderma spp., namely, T. asperellum ICC012 and T. gamsii ICC080 with the representative formulation Remedier, were provided showing significant adverse effects on reproduction at the lower doses tested. In addition, it is not possible to conclude whether effects were due to toxicity or pathogenicity, since the study duration was shorter than recommended in the test guideline OPPTS 885.4340. One open literature study with Trichoderma harzianum on soil oribatid mites was provided in which some reproductive parameters were adversely affected.

During the peer review, further information on the potential infectiveness and pathogenicity were submitted indicating potential adverse effects after exposure to Trichoderma harzianum. Although pathogenicity cannot be excluded, insufficient information was available to perform the risk assessment and to finalise the assessment of toxicity, infectivity and pathogenicity of Trichoderma afroharzianum T‐22 to non‐target arthropods other than bees. A low risk cannot be concluded for the intended uses in field and temporary protected cropping systems (e.g. Walk‐in tunnels) (data gap leading to an issue that could not be finalised, see Section 9.1). It is noted that the RMS disagreed and considered a risk is identified.

Based on the representative use indoors and in greenhouses (permanent) and in line with EFSA (2015), minimal exposure to the active substance and secondary metabolites is expected; hence, low risk can be concluded for non‐target arthropods other than bees for the uses in these production systems. The RMS disagreed and considered that, for all representative uses, secondary metabolites are expected to be of limited or no concern and a low risk can be concluded.

6.5. Soil organisms

No study is available with soil macro‐organisms other than earthworms. For earthworms, no study is available with the active substance. One 14‐day study is available with the representative formulation which does not report adverse effects.

Additional open literature studies are available with other Trichoderma spp. Adverse effects were not observed in relevant studies, and no evidence of infectivity or pathogenicity was found.

The available information is considered sufficient to finalise the assessment of infectivity and pathogenicity of Trichoderma afroharzianum T‐22 to earthworms.

Toxicity data were not available for secondary metabolites to perform a hazard characterisation (data gap leading to an issue that could not be finalised, see Section 9.1). The risk assessment for secondary metabolites could not be finalised for earthworms for the representative uses in field and temporary protected cropping systems (e.g. walk‐in tunnels) (see also Section 5.2).

Based on the representative use indoors and greenhouses (permanent) and in line with EFSA (2015), minimal exposure to secondary metabolites is expected; hence, low risk can be concluded for soil organisms for these representative uses. The RMS disagreed and considered that, for all representative uses, secondary metabolites are expected to be of limited or no concern, and a low risk can be concluded.

For soil microorganisms, relevant studies with the active substance are not available. Studies with other Trichoderma spp., namely T. asperellum ICC012, are available. Moreover, a study on the formulated product Remedier is also available. Limited, transient or no effects were reported in these studies. The available information is considered sufficient to finalise the risk assessment. A low risk can be concluded for soil microorganisms for all intended uses.

6.6. Additional information

No study is available for non‐target terrestrial plants with either the active substance or the formulation for representative uses.

During the peer review, further data on the potential infectivity and pathogenicity of Trichoderma afroharzianum T‐22 were requested. One literature study carried out with Trichoderma harzianum (now Trichoderma afroharzianum) strains T39 and T12 reporting pathogenicity to maize was identified. It should be noted that generally plant pathogens should be considered host and strain‐specific; hence, extrapolation from maize to other non‐target terrestrial plants, and from Trichoderma afroharzianum T‐22 to other strains of Trichoderma afroharzianum is uncertain. Therefore, with the available data, it cannot be concluded whether Trichoderma afroharzianum T‐22 has the potential to exert pathogenic effects on non‐target terrestrial plants.

7. OVERVIEW OF THE RISK ASSESSMENT OF THE ORGANISM OR METABOLITES OF (POTENTIAL) CONCERN LISTED IN RESIDUE DEFINITIONS TRIGGERING ASSESSMENT OF EFFECTS DATA FOR THE ENVIRONMENTAL COMPARTMENTS (TABLES 1, 2, 3, 4)

TABLE 1.

Soil.

Compound (name and/or code)	Ecotoxicology
T. afroharzianum T‐22	Low risk
Secondary metabolites of potential concern	Risk assessment not finalised for the representative uses in field and temporary protected cropping systems (e.g. walk‐in tunnels)

TABLE 2.

Groundwater. ^a

Compound (name and/or code)	> 0.1 μg/L at 1 m depth for the representative uses b Step 2	Biological (pesticidal) activity/relevance Step 3a.	Hazard identified Steps 3b. And 3c.	Consumer RA triggered Steps 4 and 5	Human health relevance
T. afroharzianum T‐22	Not assessed (quantitative limit applicable only for chemical pesticides)	Yes	–	Not assessed	Yes
Secondary metabolites of potential concern (including emodin)	Open	Open	Emodin: genotoxic potential, but levels present in the MPCP were below the LOD Other metabolites: Open	Open	Open

^a Assessment according to European Commission guidance of the relevance of groundwater metabolites (2003).

^b FOCUS scenarios or relevant lysimeter.

TABLE 3.

Surface water and sediment.

Compound (name and/or code)	Ecotoxicology
T. afroharzianum T‐22	Low risk to aquatic organisms
Secondary metabolites of potential concern	Risk assessment not finalised

TABLE 4.

Air.

Compound (name and/or code)	Toxicology
Trichoderma afroharzianum T‐22 (conidia, spores)	No evidence of toxicity, pathogenicity or infectivity in rat after acute intratracheal (LD50 > 5.8 × 107 CFU/animal) administration
6‐PP (volatile compound)	Authorised as a food flavouring substance (FL no. 10.031) by Commission Implementing Regulation (EU) No 872/2012. Adverse effects in the respiratory tract (irritation and histamine release)

8. PARTICULAR CONDITIONS PROPOSED TO BE TAKEN INTO ACCOUNT BY RISK MANAGERS

Risk mitigation measures (RMMs) identified following consideration of Member State (MS) and/or applicant's proposal(s) during the peer review, if any, are presented in this section. These measures applicable for human health and/or the environment leading to a reduction of exposure levels of operators, workers, bystanders/residents, environmental compartments and/or non‐target organisms for the representative uses are listed below. The list may also cover any RMMs as appropriate, leading to an acceptable level of risks for the respective non‐target organisms.

It is noted that final decisions on the need of RMMs to ensure the safe use of the plant protection product containing the concerned active substance will be taken by risk managers during the decision‐making phase. Consideration of the validity and appropriateness of the RMMs remains the responsibility of MSs at product authorisation, taking into account their specific agricultural, plant health and environmental conditions at national level.

8.1. Particular conditions proposed for the representative uses evaluated (Table 5)

TABLE 5.

Risk mitigation measures proposed for the representative uses assessed.

Representative use	All representative uses
Operator exposure	In the absence of a quantitative risk assessment, the use of personal and respiratory protective equipment (PPE/RPE) for the operators and workers might be considered to reduce the exposure (and the risk of adverse effects due to the sensitisation potential of microorganisms)
Worker exposure

9. CONCERNS AND RELATED DATA GAPS

9.1. Issues that could not be finalised

An issue is listed as ‘could not be finalised’ if there is not enough information available to perform an assessment, even at the lowest tier level, for one or more of the representative uses in line with the uniform principles in accordance with Article 29(6) of Regulation (EC) No 1107/2009 and as set out in Commission Regulation (EU) No 546/2011 ¹⁵ and if the issue is of such importance that it could, when finalised, become a concern (which would also be listed as a critical area of concern if it is of relevance to all representative uses).

An issue is also listed as ‘could not be finalised’ if the available information is considered insufficient to conclude on whether the active substance can be expected to meet the approval criteria provided for in Article 4 of Regulation (EC) No 1107/2009.

The following issues or assessments that could not be finalised have been identified, together with the reasons including the associated data gaps where relevant, which are reported directly under the specific issue to which they are related:

1. The non‐dietary risk assessment cannot be finalised due to the following data gap:

   1. Lack of analysis of the metabolite of potential concern pachybasin in the technical material (relevant for all representative uses, see Section 3).
2. Satisfactory information is not available to demonstrate that T. afroharzianum T‐22 does not cause adverse effects on maize such as the ones reported in the EPPO Alert List for Trichoderma afroharzianum as a causal agent of ear rot on maize (EPPO RS 2022/087; 2023/194; 2024/140, 2025).

   1. Robust evidence was not available to demonstrate that Trichoderma afroharzianum T‐22 does not cause adverse effects on maize (relevant for all representative uses, see Section 2).
3. The dietary consumer risk assessment cannot be finalised considering the identified data gaps (see Section 4).

   1. Adequate qualitative and quantitative information on the residues belonging to the anthraquinone family as metabolites of potential toxicological concern for human health (emodin and pachybasin) (relevant for all representative uses on root and tuber vegetables and on stem vegetables, see Section 4).
   2. Adequate experimental evidence on the absence/presence, persistence and possible multiplication/growth of viable cells of Trichoderma afroharzianum T‐22 on edible commodities (stem vegetables) which is relevant for potential in‐situ production of non‐viable residues (emodin and pachybasin) of human health concern following treatment according to the representative uses (relevant for all representative uses on stem vegetables, see Section 4).
4. Satisfactory information was not available on the production of secondary metabolites (see Section 5.2), their levels present in the environment after application, and their potential toxicity in order to conclude on the risk assessment for non‐target terrestrial organisms (birds, wild mammals, honey bees, non‐target arthropods, earthworms, other soil macro‐ and microorganisms) and non‐target aquatic organisms (fish, freshwater invertebrates, algae and aquatic plants), leading to an assessment not finalised (see Sections 5.2 and 6) considering the identified data gap:

   1. Pending on further investigations on the production of secondary metabolites (see Section 5.2) and their levels present in the environment after application, further considerations will have to be given to their potential toxicity (see Section 6) in order to conclude on the risk assessment for terrestrial non‐target organisms (birds, wild mammals, honey bees, non‐target arthropods, earthworms, other soil macro‐ and microorganisms), relevant for the representative field use) and aquatic organisms (fish, freshwater invertebrates, algae and aquatic plants other than algae, relevant for all representative uses).
5. Potential infectivity, pathogenicity and adverse effects to non‐target arthropods other than bees (see Section 6):

   1. Data and information for the assessment of the potential infectivity and pathogenicity to non‐target arthropods other than bees (relevant for the field and temporary protected cropping systems (e.g. walk‐in tunnel) representative uses evaluated; see Sections 5 and 6).

9.2. Critical areas of concern

An issue is listed as a critical area of concern if there is enough information available to perform an assessment for the representative uses in line with the uniform principles in accordance with Article 29(6) of Regulation (EC) No 1107/2009 and as set out in Commission Regulation (EU) No 546/2011, and if this assessment does not permit the conclusion that, for at least one of the representative uses, it may be expected that a plant protection product containing the active substance will not have any harmful effect on human or animal health or on groundwater, or any unacceptable influence on the environment.

An issue is also listed as a critical area of concern if the assessment at a higher tier level could not be finalised due to lack of information, and if the assessment performed at the lower tier level does not permit the conclusion that, for at least one of the representative uses, it may be expected that a plant protection product containing the active substance will not have any harmful effect on human or animal health or on groundwater, or any unacceptable influence on the environment.

An issue is also listed as a critical area of concern if, in the light of current scientific and technical knowledge using guidance documents available at the time of application, the active substance is not expected to meet the approval criteria provided for in Article 4 of Regulation (EC) No 1107/2009.

The following critical areas of concern are identified, together with any associated data gaps, where relevant, which are reported directly under the specific critical area of concern to which they are related.

Critical areas of concern have not been identified.

9.2.1. Overview of the concerns identified for each representative use considered (Table 6)

TABLE 6.

Overview of concerns reflecting the issues not finalised, critical areas of concern and the risks identified that may be applicable for some but not for all uses or risk assessment scenarios.

Representative use		Grass seeds (field)	Tuberous plants (field)	Stem vegetables (field/temporary protection)	Stem vegetables (indoor/greenhouses (permanent))
Operator risk	Risk identified
	Assessment not finalised	X1	X1	X1	X1
Worker risk	Risk identified
	Assessment not finalised	X1	X1	X1	X1
Resident/bystander risk	Risk identified
	Assessment not finalised	X1	X1	X1	X1
Consumer risk	Risk identified
	Assessment not finalised		X3	X3	X3
Risk to wild non‐target terrestrial vertebrates	Risk identified
	Assessment not finalised	X4	X4	X4
Risk to wild non‐target terrestrial organisms other than vertebrates	Risk identified
	Assessment not finalised	X4, 5	X4, 5	X4, 5
Risk to aquatic organisms	Risk identified
	Assessment not finalised	X4	X4	X4
Groundwater exposure to active substance	Legal parametric value breached
	Assessment not finalised
Groundwater exposure to metabolites	Legal parametric value breached a
	Parametric value of 10 μg/L b breached
	Assessment not finalised	X	X	X

Note: The superscript numbers relate to the numbered points indicated in Sections 9.1 and 9.2. Where there is no superscript number, see Sections 2– 7 for further information.

^a When the consideration for classification made in the context of this evaluation under Regulation (EC) No 1107/2009 is confirmed under Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008.

^b Value for non‐relevant metabolites prescribed in SANCO/221/2000‐rev. 10 final, European Commission (2003).

(If a particular condition proposed to be taken into account to manage an identified risk, as listed in Section 8, has been evaluated as being effective, then ‘risk identified’ is not indicated in Table 6.)

In addition to the issues indicated in Table 6 below, satisfactory information was not available to demonstrate that T. afroharzianum T‐22 does not cause adverse effects on maize as the ones reported in the EPPO Alert List for Trichoderma afroharzianum as a causal agent of ear rot on maize, leading to an issue that could not be finalised.

10. LIST OF OTHER OUTSTANDING ISSUES

Remaining data gaps not leading to critical areas of concern or issues not finalised but considered necessary to comply with the data requirements, and which are relevant for some or all of the representative uses assessed at EU level. Although not critical, these data gaps may lead to uncertainties in the assessment and are considered relevant.

These data gaps refer only to the representative uses assessed and are listed in the order of the sections:

• Information on all known differences between the modified microorganism and the parent wild‐type strains (relevant for all representative uses evaluated; see ‘The microbial active substance and the formulation for representative uses’).

• Data for ‘Trianum‐P’ for flammability and storage stability, including the content of active substance and microbial contaminants after storage (relevant for all representative uses evaluated; see Section 1).

• Information on the identity of the individual secondary metabolites produced by Trichoderma afroharzianum T‐22 is not available, as the identification of the individual metabolites belonging to the recognised classes of secondary metabolites has not been provided (relevant for all representative uses evaluated; see Sections 2 and 3).

• Applicant should demonstrate that available PEDs in soil do address plateau levels after multiple applications (relevant for all representative uses evaluated; see Section 5, that has relevance for issue not finalised number 2).

• Further information identified in the scientific peer‐reviewed open literature on potential, adverse effects on non‐target organisms (relevant for all representative uses evaluated; see data requirement 9.20).

ABBREVIATIONS

AChE

acetylcholinesterase

ADE

actual dermal exposure

AF

assessment factor

AMA

Amphibian Metamorphosis Assay

AP

alkaline phosphatase

AR

androgen receptor

AR

applied radioactivity

a.s.

active substance

AST

aspartate aminotransferase (SGOT)

AV

avoidance factor

BLAST

Basic Local Alignment Search Tool

bp

base pair

BUN

blood urea nitrogen

bw

body weight

CAS

Chemical Abstracts Service

CHO

Chinese hamster ovary cells

CI

confidence interval

CL

confidence limits

cm

centimetre

DAR

draft assessment report

DAT

days after treatment

DNA

deoxyribonucleic acid

EEC

European Economic Community

ETS

external transcribed spacer

FAO

Food and Agriculture Organization of the United Nations

FID

flame ionisation detector

FIR

food intake rate

FOB

functional observation battery

FOCUS

Forum for the Co‐ordination of Pesticide Fate Models and their Use

g

gram

GAP

Good Agricultural Practice

GC

gas chromatography

GC‐FID

gas chromatography with flame ionisation detector

GC‐MS

gas chromatography – mass spectrometry

GC‐NPD

gas chromatography with nitrogen phosphorus selective detector

GLM

Good Laboratory Practice

GM

geometric mean

h

hour(s)

ha

hectare

hL

hectolitre

HR

hazard rate

IMI

International Mycological Institute [CABI Bioscience, Eggham, UK (formerly International Mycological Institute; same as CMI)].

ISO

International Organization for Standardization

ITS

internal transcribed spacer

IUPAC

International Union of Pure and Applied Chemistry

iv

intravenous

kg

kilogram

L

litre

LD₅₀

lethal dose, median; dosis letalis media

m

metre

M

mol

MCV

mean corpuscular volume

mg

milligram

mL

millilitre

mm

millimetre (also used for mean measured concentrations)

mN

milli‐Newton

MOA

mode of action

MPCP

microbial pest control product

MS

mass spectrometry

MSDS

material safety data sheet

ng

nanogram

NOAEL

no observed adverse effect level

NOEL

no observed effect level

NPD

nitrogen–phosphorus detector

OECD

Organisation for Economic Co‐operation and Development

OM

organic matter content

Pa

pascal

PDA

Potato Dextrose Agar

PEC

predicted environmental concentration

pH

pH‐value

PIE

potential inhalation exposure

P_ow

partition coefficient between n‐octanol and water

PPE

personal protective equipment

ppm

parts per million (10⁻⁶)

ppp

plant protection product

PT

proportion of diet obtained in the treated area

PTT

partial thromboplastin time

RAR

Renewal Assessment Report

RBC

red blood cells

rDNA

ribosomal deoxyribonucleic acid

REACH

Registration, Evaluation, Authorisation of Chemicals Regulation

RPE

respiratory protective equipment

S

svedberg, S (10⁻¹³ s)

SAR

systemic acquired resistance

SCAR

sequence characterised amplified region.

SD

standard deviation

SMILES

simplified molecular‐input line‐entry system

TC

technical material

TK

technical concentrate

TWA

time‐weighted average

USDA‐ARS

U.S. Department of Agriculture, Agriculture Research Service

UV

ultraviolet

W/S

water/sediment

w/v

weight per unit volume

w/w

weight per unit weight

WBC

white blood cell

WHO

World Health Organization

ε

decadic molar extinction coefficient

λ

wavelength

μg

microgram

μm

micrometre (micron)

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2017‐00279

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NOTE/UPDATE

This scientific output, approved on 03 September 2026, supersedes the previous output published on 18 October 2013 (EFSA, 2013).

Supporting information

APPENDIX A List of end points for the active substance and the formulation for representative uses

APPENDIX A. List of end points for the active substance and the formulation for representative uses

Appendix A can be found in the online version of this output (‘Supporting Information’ section): https://doi.org/10.2903/j.efsa.2026.10323

APPENDIX B. Clarification of the wording used in Section 6

Wording used in Section 6 of this conclusion to communicate the outcome of the environmental risk assessment

Wording	Interpretation
Low risk	A low risk to the non‐target organism was indicated based on screening, Tier 1 or higher Tier assessment.
High risk	A high risk to the non‐target organism was indicated based on either Tier 1 or higher Tier assessment.
High risk not excluded	Only a screening level assessment was available, which did not show a low risk. This assessment is not considered to confirm a high risk to the non‐target organism.
Pending an identified data gap in another section, further consideration may be needed	Based on the available information, it is not known if a risk assessment for non‐target organisms is triggered.

EFSA CEP Panel (EFSA Panel on Food Contact Materials, Enzymes and Processing Aids) , Álvarez, F. , Arena, M. , Auteri, D. , Leite, S. B. , Binaglia, M. , Castoldi, A. F. , Chiusolo, A. , Colagiorgi, A. , Colas, M. , Crivellente, F. , De Lentdecker, C. , De Magistris, I. , Fait, G. , Ferilli, F. , Santonja, G. G. , Gouliarmou, V. , Ippolito, A. , Istace, F. , Kardassi, D. , … Scattareggia Marchese, A. (2026). Peer review of the pesticide risk assessment of the active substance Trichoderma afroharzianum (formerly harzianum) T‐22. EFSA Journal, 24(10), e10323. 10.2903/j.efsa.2026.10323