Pest categorisation of Colletotrichum aenigma , C. alienum , C. perseae , C. siamense and C. theobromicola

Abstract The EFSA Plant Health Panel performed a pest categorisation of Colletotrichum aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola, five clearly defined fungi of the C. gloeosporioides complex causing anthracnose. The pathogens are widely distributed in at least three continents. C. aenigma and C. siamense are reported from Italy and C. alienum from Portugal, including the Madeira Islands, with a restricted distribution. C. perseae and C. theobromicola are not known to be present in the EU. However, there is uncertainty on the status of the pathogens worldwide and in the EU because of the taxonomic re‐evaluation of the genus Colletotrichum and the lack of specific surveys. The pathogens are not included in Commission Implementing Regulation (EU) 2019/2072 and there are no reports of interceptions in the EU. With the exception of C. perseae, which has a very limited number of hosts, the other four Colletotrichum species have relatively wide host ranges. Therefore, this pest categorisation focused on those hosts for which there is robust evidence that the pathogens were formally identified by a combination of morphology, pathogenicity and multilocus sequence analysis. Host plants for planting and fresh fruits are the main entry pathways into the EU. Host availability and climate suitability factors occurring in some parts of the EU are favourable for the establishment of the pathogens. No yield losses have been reported so far in the EU but in non‐EU areas of their current distribution, the pathogens have a direct impact on cultivated hosts that are also relevant for the EU. Phytosanitary measures are available to prevent the further introduction and spread of C. aenigma, C. alienum and C. siamense into the EU as well as the introduction and spread of C. perseae and C. theobromicola. C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola satisfy the criteria that are within the remit of EFSA to assess for these species to be regarded as potential Union quarantine pests.

The new Plant Health Regulation (EU) 2016/2031, on the protective measures against pests of plants, is applying from 14 December 2019. Conditions are laid down in this legislation in order for pests to qualify for listing as Union quarantine pests, protected zone quarantine pests or Union regulated non-quarantine pests. The lists of the EU regulated pests together with the associated import or internal movement requirements of commodities are included in Commission Implementing Regulation (EU) 2019/2072. Additionally, as stipulated in the Commission Implementing Regulation 2018/2019, certain commodities are provisionally prohibited to enter in the EU (high risk plants, HRP). EFSA is performing the risk assessment of the dossiers submitted by exporting to the EU countries of the HRP commodities, as stipulated in Commission Implementing Regulation 2018Regulation /2018. Furthermore, EFSA has evaluated a number of requests from exporting to the EU countries for derogations from specific EU import requirements.
In line with the principles of the new plant health law, the European Commission with the Member States are discussing monthly the reports of the interceptions and the outbreaks of pests notified by the Member States. Notifications of an imminent danger from pests that may fulfil the conditions for inclusion in the list of the Union quarantine pest are included. Furthermore, EFSA has been performing horizon scanning of media and literature.
As a follow-up of the above-mentioned activities (reporting of interceptions and outbreaks, HRP, derogation requests and horizon scanning), a number of pests of concern have been identified. EFSA is requested to provide scientific opinions for these pests, in view of their potential inclusion by the risk manager in the lists of Commission Implementing Regulation (EU) 2019/2072 and the inclusion of specific import requirements for relevant host commodities, when deemed necessary by the risk manager.

Terms of Reference
EFSA is requested, pursuant to Article 29(1) of Regulation (EC) No 178/2002, to provide scientific opinions in the field of plant health.
EFSA is requested to deliver 53 pest categorisations for the pests listed in Annex 1A, 1B, 1D and 1E (for more details see mandate M-2021-00027 on the Open.EFSA portal). Additionally, EFSA is requested to perform pest categorisations for the pests so far not regulated in the EU, identified as pests potentially associated with a commodity in the commodity risk assessments of the HRP dossiers (Annex 1C; for more details see mandate M-2021-00027 on the Open.EFSA portal). Such pest categorisations are needed in the case where there are not available risk assessments for the EU.
When the pests of Annex 1A are qualifying as potential Union quarantine pests, EFSA should proceed to phase 2 risk assessment. The opinions should address entry pathways, spread, establishment, impact and include a risk reduction options analysis.
Additionally, EFSA is requested to develop further the quantitative methodology currently followed for risk assessment, in order to have the possibility to deliver an express risk assessment methodology. Such methodological development should take into account the EFSA Plant Health Panel Guidance on quantitative pest risk assessment and the experience obtained during its implementation for the Union candidate priority pests and for the likelihood of pest freedom at entry for the commodity risk assessment of High Risk Plants.

Additional information
The pest categorisation was initiated following the commodity risk assessment of Persea americana from Israel.

2.
Data and methodologies 2.1. Data 2.1.1. Information on pest status from NPPOs In the context of the current mandate, EFSA is preparing pest categorisations for new/emerging pests that are not yet regulated in the EU. When official pest status is not available in the European and Mediterranean Plant Protection Organization (EPPO) Global Database (EPPO, online), EFSA consults the NPPOs of the relevant MSs. To obtain information on the official pest status for Colletotrichum aenigma, C. alienum and C. siamense, EFSA has consulted the NPPOs of Italy, Portugal and Spain. The results of this consultation are presented in Section 3.2.2.

Literature search
A literature search on Colletotrichum aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola and their synonyms was conducted at the beginning of the categorisation in the ISI Web of Science bibliographic database, using the scientific name of the pest as search term. Papers relevant for the pest categorisation were reviewed, and further references and information were obtained from experts, as well as from citations within the references and grey literature.

Database search
Pest information, on host(s) and distribution, was retrieved from the EPPO Global Database, the CABI databases and scientific literature databases as referred above in Section 2.1.1.
Data about the import of commodity types that could potentially provide a pathway for the pest to enter the EU and about the area of hosts grown in the EU were obtained from EUROSTAT (Statistical Office of the European Communities).
The Europhyt and TRACES databases were consulted for pest-specific notifications on interceptions and outbreaks. Europhyt is a web-based network run by the Directorate General for Health and Food Safety (DG SANT E) of the European Commission as a subproject of PHYSAN (Phyto-Sanitary Controls) specifically concerned with plant health information. TRACES is the European Commission's multilingual online platform for sanitary and phytosanitary certification required for the importation of animals, animal products, food and feed of non-animal origin and plants into the European Union and the intra-EU trade and EU exports of animals and certain animal products. Up until May 2020, the Europhyt database managed notifications of interceptions of plants or plant products that do not comply with EU legislation, as well as notifications of plant pests detected in the territory of the Member States and the phytosanitary measures taken to eradicate or avoid their spread. The recording of interceptions switched from Europhyt to TRACES in May 2020. GenBank was searched to determine whether it contained any nucleotide sequences for Colletotrichum aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola, which could be used as reference material for molecular diagnosis. GenBank ® (www.ncbi.nlm.nih.gov/genbank/) is a comprehensive publicly available database that as of August 2019 (release version 227) contained over 6.25 trillion base pairs from over 1.6 billion nucleotide sequences for 450,000 formally described species (Sayers et al., 2020).

Methodologies
The Panel performed the pest categorisation for Colletotrichum aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola, following guiding principles and steps presented in the EFSA guidance on quantitative pest risk assessment (EFSA PLH Panel, 2018), the EFSA guidance on the use of the weight of evidence approach in scientific assessments (EFSA Scientific Committee, 2017) and the International Standards for Phytosanitary Measures No 11 (FAO, 2013).
The criteria to be considered when categorising a pest as a potential Union quarantine pest (QP) are given in Regulation (EU) 2016/2031 Article 3 and Annex I, Section 1 of the Regulation. Table 1 presents the Regulation (EU) 2016/2031 pest categorisation criteria on which the Panel bases its conclusions. In judging whether a criterion is met, the Panel uses its best professional judgement (EFSA Scientific Committee, 2017) by integrating a range of evidence from a variety of sources (as presented above in Section 2.1) to reach an informed conclusion as to whether or not a criterion is satisfied.
The Panel's conclusions are formulated respecting its remit and particularly with regard to the principle of separation between risk assessment and risk management (EFSA founding regulation (EU) No 178/2002); therefore, instead of determining whether the pest is likely to have an unacceptable impact, deemed to be a risk management decision, the Panel will present a summary of the observed impacts in the areas where the pest occurs, and make a judgement about potential likely impacts in the EU. Whilst the Panel may quote impacts reported from areas where the pest occurs in monetary terms, the Panel will seek to express potential EU impacts in terms of yield and quality losses and not in monetary terms, in agreement with the EFSA guidance on quantitative pest risk assessment (EFSA PLH Panel, 2018). Article 3 (d) of Regulation (EU) 2016/2031 refers to unacceptable social impact as a criterion for quarantine pest status. Assessing social impact is outside the remit of the Panel.

3.
Pest categorisation 3.1. Identity and biology of the pest 3.1.1. Identity and taxonomy Is the identity of the pest clearly defined, or has it been shown to produce consistent symptoms and/or to be transmissible?
The genus Colletotrichum constitutes a large monophyletic group of ascomycete fungi with more than 200 accepted species classified into at least 14 species complexes and singletons . The genus includes endophytes, saprobes and plant pathogens, with the latter being responsible for several diseases of many crops worldwide Udayanga et al., 2013). In the past, cultural and morphological characters (colour and growth rate of the colonies, size and shape of conidia and appressoria, presence or absence of setae, etc.) were used to Identity of the pest (Section 3.1) Is the identity of the pest clearly defined, or has it been shown to produce consistent symptoms and to be transmissible?
Absence/presence of the pest in the EU territory (Section 3.2) Is the pest present in the EU territory? If present, is the pest in a limited part of the EU or is it scarce, irregular, isolated or present infrequently? If so, the pest is considered to be not widely distributed. Pest potential for entry, establishment and spread in the EU territory (Section 3.4) Is the pest able to enter into, become established in, and spread within, the EU territory? If yes, briefly list the pathways for entry and spread.
Potential for consequences in the EU territory (Section 3.5) Would the pests' introduction have an economic or environmental impact on the EU territory? Available measures (Section 3.6) Are there measures available to prevent pest entry, establishment, spread or impacts?

Conclusion of pest categorisation (Section 4)
A statement as to whether (1) all criteria assessed by EFSA above for consideration as a potential quarantine pest were met and (2) if not, which one(s) were not met.   (Griessinger and Roy, 2015;EPPO, 2019) for this species is COLLSM (EPPO, online).   (Griessinger and Roy, 2015;EPPO, 2019) for this species is COLLTH (EPPO, online).

Biology of the pest
Species of the genus Colletotrichum show different lifestyles that vary between species complexes, with most species being able to sequentially switch between lifestyles (de Silva et al., 2017a). The lifestyle patterns found in Colletotrichum species can be broadly categorised as necrotrophic, hemibiotrophic, saprotrophic, latent or quiescent, and endophytic. Evidence suggests that the interaction between the host plant and the endophytic Colletotrichum species can sometimes switch from mutualistic to antagonistic or pathogenic depending on the physiological condition of the plant, host genotype and environmental conditions (de Silva et al., 2017a). Therefore, Colletotrichum species may have different interactions with their hosts and exhibit differences in their life cycles independently whether they belong to the same species complex or not (da . Occurrence of crosspathogenicity between Colletotrichum species from different hosts is also observed (Braganc ßa et al., 2016;Eaton et al., 2021).
Colletotrichum aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola have life cycles similar to those of other Colletotrichum species (Figure 1). Their life cycles include both asexual and sexual reproductive stages de Silva et al., 2017a). Infection occurs via an appressorium that develops from the germinating conidium on the plant surface, followed by turgordriven penetration of the cuticle (Deising et al., 2000) and, in some cases, also of the epidermal cells by fungal hyphae (Bailey et al., 1992). Establishment within plant tissues is aided via production by the fungus of host-induced virulence effectors (Kleemann et al., 2012;O'Connell et al., 2012). Subsequently, the pathogens enter a biotrophic phase during which they remain quiescent or latent within the host tissues until environmental conditions and host physiology become conducive for their reactivation and further development (Boufleur et al., 2020). The biotrophic life strategies adopted by Colletotrichum species may also contribute to their prominence as symptomless endophytes of living plant tissues (Lu et al., 2004;Joshee et al., 2009;Rojas et al., 2010;Yuan et al., 2011). Following the biotrophic phase, Colletotrichum spp. enter a necrotrophic phase that results in death of host plant cells and the appearance of disease symptoms. During their active growth in the plant tissues, the pathogens produce acervuli (asexual fruiting structures) with masses of mucilage-embedded conidia ( Figure 1). The mucilaginous matrix is composed of glycoprotein and germination inhibitors that protect conidia against desiccation and toxins produced by the host defence mechanism (Leite and Nicholson, 1992). The conidia are dispersed over relatively short distances by water (rain, irrigation), wind-driven rain (de Silva et al., 2017a). Dispersal is also possible by insects (Gasparoto et al., 2017). They are produced on the infected host tissues throughout the season resulting in polycyclic disease cycles. The sexual stage of many Colletotrichum species, including C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola, has been observed in in vitro cultures on synthetic media but not under field conditions (Jayawardena et al., 2021).
Although it has not been documented, seeds of host plants are possibly one of the main sources of primary inoculum for the above-mentioned five Colletotrichum species, similarly to other Colletotrichum species .  Silva et al., 2017a) Like other Colletotrichum species, host infection by C. aenigma, C. alienum, C. perseae, C. siamense or C. theobromicola depends on different factors including humidity, temperature, host physiology and inoculum level (Freeman et al., 1998). In general, warm, wet or humid environmental conditions favour host infection by Colletotrichum species. According to Zhang et al. (2020a,b), the optimum temperature for the in vitro mycelial growth of C. aenigma and C. siamense was 28°C. At 36°C, no mycelial growth of C. aenigma was observed, whereas C. siamense proved to be more tolerant to temperatures higher than 36°C suggesting the potential threat posed by this species to hosts grown in areas with hot and rainy weather.

Host range/Species affected
With the exception of C. perseae, which, so far, has been reported to affect a very limited number of hosts, the other four Colletotrichum species, i.e. C. aenigma, C. alienum, C. siamense and C. theobromicola, have relatively wide host ranges (see Appendix A). It should be noted that, in some cases, more than one of the above-mentioned five Colletotrichum species were identified to be associated with anthracnose on a single host, whereas in other cases, other species of the C. gloeosporioides complex or of other Colletotrichum species complexes were also involved Sharma et al., 2017;Yokosawa et al., 2017;Fu et al., 2019;Chen et al., 2020;Zhang et al., 2020a,b). The host range of each of the five Colletotrichum species and particularly that of C. perseae, which has been identified recently , might be wider than that currently reported as, in the past, when molecular tools were not available, Colletotrichum species identified as C. gloeosporioides sensu lato based on morphology and pathogenicity, might have belonged to one of the above-mentioned species.
Given that Colletotrichum species are commonly found on many plant species as pathogens, endophytes and occasionally as saprobes, and that their accurate identification and their discrimination from other closely-related Colletotrichum species is only possible by using molecular tools, this Pest categorisation will focus on those hosts for which there is robust evidence in the literature that (i) the pathogens were isolated and identified by both morphology and multilocus gene sequencing analysis, (ii) the Koch's postulates were fulfilled through pathogenicity tests performed on unwounded plant tissues, and (iii) their impacts on crop yield were documented. The reported hosts in the literature of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola that fulfil the above-mentioned criteria are considered by the Panel as main hosts and are listed in Table 2. Appendix B provides an overview on the main hosts which can be infected or co-infected by more than one of the five Colletotrichum species.

Intraspecific diversity
The sexual stage of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola has been observed in in vitro cultures but not under field conditions. However, other species of the C. gloeosporioides complex form perithecia (sexual fruiting bodies) on their hosts (Dowling et al., 2020). The ability of Colletotrichum species to differentiate sexual reproductive stages enhances their genomic plasticity and adaptation to various and/or adverse environmental conditions, including the selection of fungicide-resistant populations. It is generally acknowledged that the risk of fungicide resistance development increases when sexual recombination occurs in the life cycle (FRAC, 2014). With this respect, many isolates of C. siamense from commercial peach orchards in South Carolina (USA) were found to be resistant to quinone outside inhibitors (QoI) fungicides and some even dual resistant to QoI and benzimidazole fungicides .
No information exists in the literature on differences in aggressiveness among isolates of each of the five Colletotrichum species, although such differences have been reported for other species of the C. gloeosporioides complex .

Detection and identification of the pest
Are detection and identification methods available for the pest?
Yes, methods for the detection and identification of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola and their discrimination from other closely related Colletotrichum species are available.
Plants infected by C. aenigma, C. alienum, C. perseae, C. siamense or C. theobromicola show symptoms of anthracnose, which may include dark brown stem and fruit spots, stem cankers, pre-and post-harvest fruit rot, leaf spots and wilt, shoot-tip dieback and defoliation (Rodrigues et al., 2014; Diao et al., 2017;Sharma et al., 2017Sharma et al., , 2022de Silva et al., 2017b;Giblin et al., 2018;Chethana et al., 2019;Costa et al., 2019;Chaves et al., 2020;Chen et al., 2020;Chung et al., 2020;Mao et al., 2020;Yokosawa et al., 2020;Andrioli et al., 2021;Carbone et al., 2021;Luo et al., 2021;Moral et al., 2021;Han et al., 2022). However, these symptoms are similar to those caused by other Colletotrichum species belonging either to the C. gloeosporioides complex or to other Colletotrichum species complexes. If fruiting structures (acervuli with conidia and/or perithecia with ascospores) are detected on the symptomatic plant tissues using a magnifying lens, they are similar to those of other Colletotrichum species. It should be also noted that during the biotrophic phase, the pathogens remain quiescent or latent within the host tissues until environmental conditions and host physiology become conducive for their reactivation and the development of disease symptoms (see Section 3.1.2 Biology of the pest). Based on the above, it is unlikely that C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola could be detected based only on visual inspection of their host plants.
The pathogens can be readily isolated on culture media and description of their cultural and morphological characteristics is available in the literature Rojas et al., 2010;Weir et al., 2012;Sharma et al., 2017;Hassan et al., 2018;Fu et al., 2019;Ahmad et al., 2021). However, as some of these characteristics are similar to or overlap with those of other Colletotrichum species, and moreover, they vary under changing environmental conditions Liu et al., 2016), the pathogens cannot be reliably identified based only on morphology. A polyphasic approach, combining the application of molecular methods, such as multilocus gene sequencing analysis with morphological and pathogenicity data, is currently recognised as being the most reliable method for the identification of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola and their discrimination from other closely related Colletotrichum species Weir et al., 2012). More specifically, C. aenigma can be distinguished from other closely related Colletotrichum species based on sequence analysis of tub2 and gs genes ; C. alienum using cal or gs genes ; C. perseae can be well resolved with sequence analysis of ApMAT and gs genes ; C. siamense is distinguished by cal or tub2 gene sequence data and C. theobromicola by ITS sequences . Nucleotide sequences of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola are available in GenBank (www.ncbi.nlm.nih.gov/genbank) and could be used as reference material for molecular diagnosis.
No EPPO Standards are available for the detection and identification of C. aenigma, C. alienum, C. perseae, C. siamense or C. theobromicola.

3.2.
Pest distribution 3.2.1. Pest distribution outside the EU The current geographical distribution of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola outside the EU is shown in Table 3 and Figures 2-6. The records are based on CABI Crop Protection Compendium (online; accessed on 15/2/2022), Farr and Rossman (online; https://nt. ars-grin.gov/fungaldatabases/; accessed on 15/2/2022) and other sources (published articles until May 2022) as well as on whether the species is included in the EPPO Global Database (online; last accessed on 15/2/2022). Details of the current geographical distribution of each of the above-mentioned pathogens outside the EU are presented in Appendix C.
There is uncertainty with respect to the actual geographical distribution of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola outside the EU, as in the past, when molecular tools (i.e. multigene phylogenetic analysis) were not available, the pathogens might have been misidentified based on morphology and pathogenicity tests only, which cannot reliably identify them.

Pest distribution in the EU
Is the pest present in the EU territory? If present, is the pest in a limited part of the EU or is it scarce, irregular, isolated or present infrequently? If so, the pest is considered to be not widely distributed.
Colletotrichum aenigma, C. alienum and C. siamense have been reported to be present in the EU, with a restricted distribution. More specifically, C. aenigma and C. siamense have been reported from Italy and C. alienum from Portugal, including Madeira Islands. There are no reports of C. perseae and C. theobromicola being present in the EU.
According to Schena et al. (2014), C. aenigma was isolated from Citrus sinensis, Pyrus communis and Olea europaea in Italy during the period 1992-2011 and was tested for its pathogenicity on detached olive fruits (cv. Coratina). Results showed that C. aenigma was a weak pathogen on green olives, but it induced noticeable rot on olives at the colour changing stage (ripening olives). In Mosca et al. (2014) studies, one isolate of C. aenigma from P. communis originated in Apulia (Italy) was used as reference material. According to the NPPO of Italy, the only report of C. aenigma in Italy is on Olea europaea; the NPPO further noted that the fungus is a weak pathogen with sporadic presence.
C. alienum has been reported from Portugal, including Madeira Islands. More specifically, in their studies, Liu et al. (2013)  Far and Rossman (online; https://nt.ars-grin.gov/fungaldatabases/) reported C. alienum as being present in Spain and cited Crous et al. (2013). However, this is an unreliable record as, in none of the primary literature sources , C. alienum is reported as present in Spain. The NPPO of Spain stated that C. alienum is not known to be present in Spain.
In a study on the biodiversity of fungi on Vitis vinifera (grapevine), Jayawardena et al. (2018) isolated among other fungi, C. siamense from grapevines grown in the Forl ı-Cesena Province, Italy. However, the pathogenicity of the fungus on grapevine was not investigated. According to the NPPO of Italy, C. siamense has been isolated in 2021 from receptive stigmas of walnut fruit but compared to other Colletotrichum species (C. fioriniae and C. nymphaeae), to date it is not pathogenic on walnut in Italy. C. perseae and C. theobromicola have not been reported from the EU territory. There is uncertainty with respect to the actual distribution of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola in the EU, as in the past, when molecular tools (i.e. multigene phylogenetic analysis) were not available, the pathogens might have been misidentified based on morphology and pathogenicity tests, which cannot reliably identify them.

Hosts or species affected that are prohibited from entering the Union from third countries
A list of hosts included in Annex VI of Commission Implementing Regulation (EU) 2019/2072 is provided in Table 4. Some of the hosts which belong to the genera Acacia, Annona, Diospyros, Jasminum, Juglans, Persea, Prunus, Persea as well as Ficus carica, are included in the Commission Implementing Regulation (EU) 2018/2019 on high-risk plants.

3.4.
Entry, establishment and spread in the EU

Entry
Is the pest able to enter into the EU territory? If yes, identify and list the pathways.
Yes. C. aenigma, C. alienum and C. siamense have already entered the EU and they may further enter via the host plants for planting and the fresh fruit pathways. Similarly, C. perseae and C. theobromicola could potentially enter the EU territory via the host plants for planting and the fresh fruit pathways.
Comment on plants for planting as a pathway.
Host plants for planting is a main pathway for the entry of the pathogens into the EU territory.
The Panel identified the following main pathways for the entry of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola into the EU territory: 1) host plants for planting, and 2) fresh fruit of host plants, originating in infested third countries (Table 3) The pathogens could potentially enter the EU territory on nuts, cut flowers and plant parts of their hosts for ornamental or medicinal purposes. However, these are considered minor pathways for the entry of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola into the EU.
Although seeds are reported as one of the primary sources of inoculum for many Colletotrichum species, there is no evidence of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola being transmitted by seeds of their host plants. Therefore, uncertainty exists on seeds of host plants as a pathway for the entry of the above-mentioned five Colletotrichum species into the EU.
No information specific for C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola exists in the available literature on their potential to survive in soil, but in general, Colletotrichum species appear to be poor competitors in soil (see Section 3.1.2 Biology of the pest). Therefore, uncertainty exists on the soil and other substrates associated or not with host and non-host plants for planting as a pathway of entry of the pathogens into the EU territory.
C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola are unlikely to enter the EU by natural means (rain, wind-driven rain, insects, etc.) because of the long distance between the infested third countries and the EU Member States. Although there are no quantitative data available, spores of the pathogens may be also present as contaminants on other substrates or objects (e.g. non-host plants, second hand agricultural machinery and equipment, crates, etc.) imported into the EU. Nevertheless, these are considered minor pathways for the entry of the pathogens into the EU territory.  (Table 4). Among the third countries from where the introduction of the above-mentioned plant material is not prohibited are Turkey and UK, which have been reported to be infested with C. siamense and C. aenigma, respectively (see Section 3.2.1). • Annex VI (9) prohibits the introduction of plants for planting of Malus Mill., Prunus L., Pyrus L. and Fragaria L., other than seeds, from certain third countries ( for the introduction into the Union from certain third countries of fruits (fresh or chilled) of Actinidia Lindl., Annona L., Carica papaya L., Diospyros L., Fragaria L., Malus L., Mangifera L., Persea americana Mill., Prunus L., Pyrus L. and Vitis L. (Table 4).
Among the third countries from which a phytosanitary certificate is not required, Turkey has been reported to be infested with C. siamense (see Section 3.2.1).
• Annex XI, Part A (5) requires phytosanitary certificate for the introduction into the Union from certain third countries of fruits of Punica granatum L. (Table 4). Among the third countries from which a phytosanitary certificate is not required, are countries which have been reported to be infested with the pathogens (see Section 3.2.1). Among the third countries from which a phytosanitary certificate is not required, Turkey has been reported to be infested with C. siamense.

Leaves of host plants
• Annex XI, Part A (3) requires phytosanitary certificate for the introduction into the Union from third countries other than Switzerland of Camellia spp. L. plant parts, other than fruits and seeds It should be noted that among the host plant genera included in Table 5 The volume of fresh produce of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola main hosts originated in infested third countries and imported into the EU territory during the period 2016-2020 is presented in Table 6. Appendix D provides import statistics for individual third countries.  Notifications of interceptions of harmful organisms began to be compiled in Europhyt in May 1994 and in TRACES in May 2020. No records of interceptions by EU Members States specific for C. aenigma, C. alienum, C. perseae, C. siamense or C. fructicola exist in Europhyt (accessed on 6 September 2022). Nevertheless, until May-June 2020, there have been 21 interceptions of unidentified at species level Colletotrichum. No records of any of the five Colletotrichum species exist in TRACES database since May 2020 (accessed on 15/5/2022). However, there is only one report of C. acutatum.

Establishment
Is the pest able to become established in the EU territory?
Yes. C. aenigma, C. alienum and C. siamense are present in the EU, which indicates that both the biotic (host availability) and abiotic (climate suitability) factors occurring in parts of the EU are also favourable for the establishment of the other two species, i.e. C. perseae and C. theobromicola.
Given their biology, the five Colletotrichum species could potentially be transferred from the pathways of entry to the host plants grown in the EU via splash-dispersed spores, contaminated soil and other plant growth substrates associated with plants for planting, and rain or irrigation water. The frequency of this transfer will depend on the volume and frequency of the imported commodities, their destination (e.g. nurseries, retailers, packinghouses) and its proximity to the hosts grown in the EU territory, as well as on the management of plant residues and fruit waste.
Climatic mapping is the principal method for identifying areas that could provide suitable conditions for the establishment of a pest taking key abiotic factors into account (Baker et al., 2000). Availability of hosts is considered in Section 3.4.2.1. Climatic factors are considered in Section 3.4.2.2.

EU distribution of main host plants
As noted above and shown in Appendix A, except for C. perseae, whose host range is limited so far to pepper, olive, avocado and grapevine, the other four Colletotrichum species, i.e. C. aenigma, C. alienum, C. siamense and C. theobromicola have relatively wide host ranges. In addition, most of the main hosts of the above-mentioned five Colletotrichum species (Table 2) are widely distributed in the EU territory, in commercial production (fields, orchards, greenhouses) and in home gardens. The harvested area of most of the main hosts of each of the above-mentioned Colletotrichum species cultivated in the EU 27 in recent years is shown in Table 7. Appendix E provides production statistics for individual Member States.  Of the five Colletotrichum species, C. aenigma and C. perseae have been reported from three continents (i.e. C. aenigma from America, Asia and Europe and C. perseae from America, Asia and Oceania), C. theobromicola from four continents (i.e. Africa, America, Asia and Oceania) and C. alienum and C. siamense from all the five continents (i.e. Africa, America, Asia, Europe and Oceania).
The global K € oppen-Geiger climate zones (Kottek et al., 2006) describe terrestrial climate in terms of average minimum winter temperatures and summer maxima, amount of precipitation and seasonality (rainfall pattern).
Based on the data available in the literature on the exact locations of the infested areas, C. aenigma has been reported from areas with BSh, BSk, Cfa, Cfb, Csa, Dfb and Dfc climate zones; C. alienum from areas with BSh, BSk, Cfa, Cfb, Cfc, Csa, Csb and Dfc climate zones; C. perseae from areas with BSh, Cfa, Cfb, Csa and Dfb climate zones; C. siamense from areas with BSh, BSk, Cfa, Cfb, Cfc, Csa, Csb, Csc, Dfb and Dfc climates and C. theobromicola from areas with BSh, BSk, Cfa, Cfb, Cfc, Csa, Dfb and Dfc climates. The above-mentioned climate zones, where each of those five Colletotrichum species is currently present, are comparable to those occurring in parts of the EU territory where hosts are also grown (Figures 7-11).
Therefore, it can be concluded that the climatic conditions occurring in some parts of the EU territory are favourable for the establishment of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola. However, uncertainty exists on whether the pathogens could potentially establish in EU areas belonging to other than the climate zones shown in Figures 7-11, where hosts are also present.

Spread
Describe how the pest would be able to spread within the EU territory following establishment?
Following establishment, Colletotrichum aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola could potentially spread within the EU territory by natural and human-assisted means.
Host plants for planting is one of the main means of spread of the pathogens within the EU territory.
Following their introduction into the EU territory, C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola, similarly to other Colletotrichum species, could potentially spread via natural and human-assisted means.
Spread by natural means. Colletotrichum species can spread locally mainly by water (rain, irrigation) (Madden et al., 1996;Freeman et al., 2002;Mouen Bedimo et al., 2007;Penet et al., 2014). Winddriven rain and insects may also contribute to the dispersal of Colletotrichum spp. spores (Gasparoto et al., 2017). In some pathosystems (e.g. C. acutatum sensu stricto and C. gloeosporioides sensu stricto affecting citrus), spread of the pathogens may also occur via wind-disseminated ascospores (Silva-Junior et al., 2014). However, there is uncertainty on the potential of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola to spread via wind-borne ascospores, as the presence of their sexual stage has not been reported so far under field conditions (see Section 3.1.2 Biology of the pest).
Spread by human-assisted means. The pathogens can spread over long distances via the movement of infected host plants for planting (rootstocks, grafted plants, scions, etc.), including dormant plants, as well as fresh fruits, contaminated agricultural machinery, tools, irrigation, etc.
Uncertainty exists on the potential of the pathogens to spread via the seeds of their host plants and soil or other plant growth substrates, due to lack of evidence.

Impacts
Would the pests' introduction have an economic or environmental impact on the EU territory?
Yes, the introduction of, C. perseae, C. theobromicola and the further introduction of Colletotrichum aenigma, C. alienum and C. siamense in the EU is likely to have yield and quality impacts in some parts of the territory. Nevertheless, the magnitude of the impacts is not known, especially in cases where more than one of the above-mentioned Colletotrichum species would coinfect a single host.
Species of the genus Colletotrichum are known to infect several economically important cultivated tropical, subtropical and temperate fruit crops, vegetables and ornamentals, causing severe damage and, consequently, resulting in significant losses (Bailey and Jeger, 1992;Lima et al., 2011;Cannon et al., 2012;Anderson et al., 2013;Guarnaccia et al., 2016;de Silva et al., 2017b).
In the areas of their current distribution, C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola are reported to cause anthracnose and pre-and post-harvest fruit rots on their host plants (see Section 3.1.5). Andrioli et al. (2021) reported that in Brazil, early infection of sweet persimmon fruit by anthracnose caused by C. aenigma, C. asianum, C. fructicola and C. nymphaeae caused premature fruit drop. Moreover, the disease developed further during fruit ripening and after harvest, leading to post-harvest losses of 50-90%. In China, anthracnose of strawberry caused by C. aenigma, C. siamense, C. fructicola and C. gloeosporioides sensu stricto was responsible for nearly 50% of seedlings necrosis and > 40% of production losses in nurseries as well as for up to 80% yield losses . Chung et al. (2020) reported that in Taiwan from 2010 to 2016, C. siamense together with C. karstii, C. fructicola and C. boninense was responsible for a 30-40% loss of strawberry seedlings and of approximately 20% loss of plants after transplanting. In Hubei province, China, an incidence of 45% of strawberry crown rot caused by C. siamense was reported by Luo et al. (2021). Gimcheon, South Korea (Kim et al., 2021); the most severely affected vineyards showed a disease incidence on grape berries of up to 50% with the infected berries displaying sunken necrotic lesions covered by orange conidial masses. In 2020, a 5% fruit damage caused by C. aenigma and C. perseae was estimated on pepper crops in two different locations in Southern (Fatsail) and Central Israel (Sde Warburg), respectively (Sharma et al., 2022). At both locations, anthracnose symptoms were observed only on pepper fruits and not on leaves or stems. According to Wang et al. (2020c), C. aenigma was the causal agent of a serious anthracnose disease of walnut orchards in Xingtai Hebei, China. Disease symptoms included brown to black circular or irregular sunken lesions on walnut fruits, with an incidence of 31-41% and circular to irregular brown to grey lesions on leaves, with an incidence of 1-2%. Additionally, Wang et al. (2017) identified C. siamense as the causal agent of walnut anthracnose in Shandong Province, China, which resulted in 50% yield loss. Ahmad et al. (2021) reported that in 2019, 30% of mango fruits at different markets of the Fengtai district, Beijing, China, exhibited severe typical symptoms of anthracnose caused by C. alienum. Li et al. (2019) identified C. siamense as the most dominant among 13 Colletotrichum species causing anthracnose on mango crops in the Provinces of Hainan, Yunnan, Sichuan, Guizhou, Guangdong and Fujian of Southern China. The same authors reported that the annual yield loss because of the disease was 30-60% reaching 100% under favourable climatic conditions.

C. aenigma was identified to be the causal agent of anthracnose outbreaks in different vineyards in
Anthracnose is the most devastating disease of olive in Uruguay (Leoni et al., 2018), particularly in orchards located in areas characterised by frequent high relative humidity and rainfall (around 1,100 mm per year). During the last 10 years, those areas were massively planted with olives for oil production using an intensive rainfed plantation system, which favours anthracnose development. According to Moreira et al. (2021), since 2017, severe anthracnose outbreaks have been observed in those areas leading to high yield losses and decreased olive oil quality (increased acidity and decreased organoleptic properties). C. alienum and C. theobromicola were identified as the causal agents of those outbreaks together with C. acutatum sensu stricto, C. nymphaeae and C. fioriniae of the C. acutatum complex .
Avocado is a high value crop grown in tropical and subtropical areas worldwide. Under the subtropical Mediterranean conditions of Israel, avocado fruit that set during the winter are seriously affected by post-harvest anthracnose which causes significant reduction in their shelf-life and marketability (Freeman et al., 1998). Sharma et al.'s (2017) studies showed that nine Colletotrichum species among which C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola were involved in anthracnose disease of avocado in Israel. In Chile, anthracnose has increased during the last decades along with the establishment of new avocado orchards in humid areas. A survey carried out by Bustamante et al. (2020) in four commercial orchards located in the regions of Valparaiso, Metropolitana and O'Higgins (Chile) revealed that C. perseae and nine more Colletotrichum species belonging to four different Colletotrichum species complexes were associated with avocado anthracnose.
Although no quantitative data are available, C. alienum has been identified as the most economically important pathogen of Proteaceae in Australia, Europe and South Africa .
In India, chilli anthracnose caused by both C. siamense and C. fructicola was reported to adversely affect the quality of chilli fruits resulting in significant yield losses and reduced marketability . Similarly, Oo et al. (2021) reported that C. siamense caused typical anthracnose symptoms on approximately 15-20% of chilli fruits (cv. Manita) growing in Goesan County, Chungcheong province, South Korea. C. siamense and C. truncatum were identified as the causal agents of severe anthracnose outbreaks in onion crops in southwest India (Chowdappa et al., 2015). P erez-Mora et al. (2020) showed that in northern Sinaloa, Mexico, C. siamense was the only Colletotrichum species causing anthracnose symptoms (petal necrosis, fruit lesions) on Mexican lime (C. aurantifolia) resulting in high crop losses and unmarketable fruits. C. siamense was reported to cause a 50-90% incidence of anthracnose on red-fleshed apples (Malus niedzwetzkyana) in commercial orchards in Shandong province, China (Han et al., 2022).
Nine Colletotrichum species among which C. siamense and C. theobromicola were found to be associated with anthracnose of Annona spp., the most important disease of annonaceous crops in Brazil, causing yield losses of up to 70% particularly in periods of extended rainfall during the flowering and fruit developmental stages . It should be noted that A. squamosa (sugar apple) and A. muricata (soursop) have been arousing great interest in the international market for their fresh and processed fruit as well as for the production of biocomposites of medicinal, allelopathic or pesticide importance (Lemos, 2014). According to Veloso et al. (2018), C. siamense was the most dominant among seven Colletotrichum species causing anthracnose on cashew in Brazil with more than 40% yield losses.
Citrus anthracnose caused by Colletotrichum spp. is a serious disease limiting production globally. Preharvest anthracnose reduces yield, while post-harvest anthracnose affects fruit quality, negatively impacting fruit export and marketability (Phoulivong et al., 2012). During a survey conducted in citrus orchards severely affected by anthracnose in Australia (Victoria, New South Wales, Queensland), Wang et al. (2021) identified six Colletotrichum species as the causal agents among which C. siamense and C. theobromicola. C. siamense was also reported to cause anthracnose of papaya fruit in China with an average disease incidence of 30% and over 60% in some orchards (Zhang et al., 2021a,b,c). According to Pardo- De la Hoz et al. (2016), six Colletotrichum species of the C. gloeosporioides complex, among which C. siamense and C. theobromicola and three species of the C. boninense complex were associated with up to 60% of yield losses in mango plantations in the state of Tolima, Colombia.
It should be noted that, in cases where anthracnose disease on a single host was reported to be associated with more than one of the five Colletotrichum species (i.e. C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola) or in cases where other species of the C. gloeosporioides complex or of other Colletotrichum species complexes were also involved, the individual contribution of C. aenigma, C. alienum, C. perseae, C. siamense or C. theobromicola to the overall impact was not determined Sharma et al., 2017;Yokosawa et al., 2017;Fu et al., 2019;Chen et al., 2020;Zhang et al., 2020a,b).
Based on the above, it is expected that the introduction of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola into the EU territory would potentially cause yield and quality losses in parts of the risk assessment area where susceptible hosts are grown. However, neither the magnitude of this impact is known nor whether the agricultural practices and chemical control measures currently applied in the EU could potentially reduce the impact of the pathogens' introduction. It is worth mentioning that, although C. aenigma and C. siamense are reported from Italy and C. alienum from Portugal, including Madeira Islands, no crop losses have been reported so far.

Available measures and their limitations
Are there measures available to prevent pest entry, establishment, spread or impacts such that the risk becomes mitigated?
Yes. Although not specifically targeted against C. aenigma, C. alienum, C. perseae, C. siamense or C. theobromicola, existing phytosanitary measures (see Sections 3.3.2 and 3.4.1) mitigate the likelihood of the pathogens' entry into the EU territory. Potential additional measures also exist to further mitigate the risk of entry and spread of the pathogens in the EU (see Section 3.6.1).

Identification of potential additional measures
Phytosanitary measures (prohibitions) are currently applied to some hosts of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola, (see Section 3.3.2). Potential additional control measures are listed in Table 8. Additional potential risk reduction options and supporting measures are shown in Sections 3.6.1.1 and 3.6.1.2.

Additional potential risk reduction options
Potential additional control measures are listed in Table 8.
Colletotrichum aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola: Pest categorisation Require pest freedom Plants, plant products and other objects come from a pest-free country or a pest-free area or a pest-free place of production.

Entry/Spread
Growing plants in isolation The use of transplants raised from pathogen-free propagation material, as well as growing transplants in weed-free areas and away from other crops that are known hosts of the pathogens may represent an effective control measure.

Entry/Spread
Managed growing conditions Anthracnose disease is generally more severe in tropical and subtropical countries. Hot and humid environmental conditions support the spread of Colletotrichum spp. Therefore, proper field drainage, plant distancing, cutting of pruning debris into small pieces for faster decomposition and removal of severely infected plants in the field or in the greenhouse represent effective strategies to manage anthracnose.

Entry/Spread/Impact
Crop rotation, associations and density, weed/volunteer control Crop rotation (wherever feasible) and control of volunteer plants may also represent effective means to reduce inoculum sources and potential survival of the pathogens on alternative hosts.

Establishment/Spread/ Impact
Roguing and pruning Infection of host plants by the pathogens usually occurs from conidia formed on infected plants or plant debris which can act as inoculum sources. These propagules are dispersed from the infected plant parts and debris to healthy plants by rain splash, free water or high humidity. To reduce the sources of inoculum, pruning of the infected by the pathogens plant parts is highly recommended.

Spread/Impact
Biological control and behavioural manipulation Some antagonistic fungi and bacteria have been tested in vitro for the biological control of the pathogens, but none of them was effective under field conditions.

Impact
Chemical treatments on crops including reproductive material Several effective fungicides are available to control anthracnose-causing species of Colletotrichum. Copper compounds, triazoles and strobilurins are effective in field treatment as well as when applied on reproductive material. The possibility of selection of fungicideresistant populations to triazoles and strobilurins must be considered.

Establishment/Spread/ Impact
Chemical treatments on consignments or during processing Copper compounds, triazoles and strobilurins are effective as post-harvest treatments against Colletotrichum species causing anthracnose and postharvest fruit rot. Calcium chloride is reported to improve the shelf-life and quality of fruits that are known hosts of anthracnose pathogens. The possibility of selection of fungicide-resistant populations should not be ruled out.

Entry/Spread
Colletotrichum aenigma, C. alienum, C. perseae, C. siamense and C. Irradiation, mechanical cleaning (brushing, washing), sorting and grading and removal of diseased plant parts could be adopted on consignment or during processing of susceptible host plants or fruit. In the packinghouse, proper sanitation practices (e.g. good drainage systems to channel out wastewater or sewage during on-farm fruit disinfection) should be built and regularly cleaned.

Entry/ Spread
Cleaning and disinfection of facilities, tools and machinery Cleaning, disinfection and disinfestation (sanitation) of equipment and facilities (including premises, storage areas) are good cultural and handling practices employed in the production and marketing of any commodity and may contribute to mitigate likelihood of entry or spread of Colletotrichum species.

Entry/Spread
Limits on soil Limits on soil are an efficient measure. Entry/Spread

Soil treatment
Although no specific studies are available on C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola, it is likely that the pathogens could potentially survive in infected plant debris in soil, similarly to other Colletotrichum species. Therefore, soil and substrate disinfection with chemical or physical (heat, soil solarisation) means represents a suitable option for control.

Use of noncontaminated water
Although Colletotrichum species could potentially spread via contaminated irrigation water, physical or chemical treatment of irrigation water is likely not to be feasible.

Spread/Impact
Waste management Treatment of the waste (deep burial, composting, incineration, chipping, production of bio-energy. . .) in authorised facilities and official restriction on the movement of waste.

Heat and cold treatments
Hot water treatment at temperatures of 50-60°C for 5-60 min, depending on the host tolerance, may be applied to reduce the likelihood of infestation of the pathogens in susceptible plants or plant organs. The combination of hot water and calcium chloride may increase the efficacy of the treatment. As Colletotrichum spp. are adapted to high temperatures, cold treatment could also mitigate infection of consignments by the pathogens.

Entry/Spread
Conditions of transport Specific requirements for mode and timing of transport of commodities to prevent escape of the pest and/or contamination.

a) physical protection of consignment b) timing of transport/trade
If plant material, potentially infected or contaminated with Colletotrichum spp. has to be transported (including proper disposal of infested waste material), specific transport conditions (type of packaging/ protection, time of transport, transport means) should be defined to prevent the pathogens from escaping. These may include, albeit not exclusively: physical protection; removal of leaves and peduncles from fruit commodities; sorting prior to transport, sealed packaging, etc.

Additional supporting measures
Potential additional supporting measures are listed in Table 9.

Post-entry quarantine and other restrictions of movement in the importing country
This information sheet covers post-entry quarantine (PEQ) of relevant commodities; temporal, spatial and end-use restrictions in the importing country for import of relevant commodities; prohibition of import of relevant commodities into the domestic country. 'Relevant commodities' are plants, plant parts and other materials that may carry pests, either as infection, infestation or contamination.

Inspection and trapping
Inspection is defined as the official visual examination of plants, plant products or other regulated articles to determine if pests are present or to determine compliance with phytosanitary regulations (ISPM 5). The effectiveness of sampling and subsequent inspection to detect pests may be enhanced by including trapping and luring techniques. As the symptoms caused by C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola on their hosts are similar to those of other anthracnose causing Colletotrichum species on the same hosts, it is unlikely that the pathogens could be detected at species level based on visual inspection only.

Entry/Establishment/Spread
Laboratory testing Examination, other than visual, to determine if pests are present using official diagnostic protocols. Diagnostic protocols describe the minimum requirements for reliable diagnosis of regulated pests.
Laboratory testing based on morphological characters and multilocus gene sequencing analysis is required for the detection and reliable identification of the pathogens.

Entry/Spread
Sampling According to ISPM 31, it is usually not feasible to inspect entire consignments, so phytosanitary inspection is performed mainly on samples obtained from a consignment. It is noted that the sampling concepts presented in this standard may also apply to other phytosanitary procedures, notably selection of units for testing.

Entry/Spread
Colletotrichum aenigma, C. alienum, C. perseae, C. siamense and C. For inspection, testing and/or surveillance purposes the sample may be taken according to a statistically based or a non-statistical sampling methodology.
Necessary as part of other risk reduction options Phytosanitary certificate and plant passport An official paper document or its official electronic equivalent, consistent with the model certificates of the IPPC, attesting that a consignment meets phytosanitary import requirements (ISPM 5) a) export certificate (import) b) plant passport (EU internal trade) Recommended for plant species known as hosts of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola

Certified and approved premises
Mandatory/voluntary certification/approval of premises is a process including a set of procedures and of actions implemented by producers, conditioners and traders contributing to ensure the phytosanitary compliance of consignments. It can be a part of a larger system maintained by the NPPO in order to guarantee the fulfilment of plant health requirements of plants and plant products intended for trade. Key property of certified or approved premises is the traceability of activities and tasks (and their components) inherent the pursued phytosanitary objective. Traceability aims to provide access to all trustful pieces of information that may help to prove the compliance of consignments with phytosanitary requirements of importing countries.
Certified and approved premises reduce the likelihood of the plants and plant products originating in those premises to be infected by the pathogens

Entry/Spread
Certification of reproductive material (voluntary/official) Plants come from within an approved propagation scheme and are certified pest free (level of infestation) following testing; Used to mitigate against pests that are included in a certification scheme The risk of entry and/or spread of the abovementioned Colletotrichum species is reduced if host plants for planting, including seeds for sowing, are produced under an approved certification scheme and tested free of these pathogens.

Entry/Spread
Delimitation of Buffer zones ISPM 5 defines a buffer zone as 'an area surrounding or adjacent to an area officially delimited for phytosanitary purposes in order to minimise the probability of spread of the target pest into or out of the delimited area, and subject to phytosanitary or other control measures, if appropriate' (ISPM 5). The objectives for delimiting a buffer zone can be to prevent spread from the outbreak area and to maintain a pest-free production place (PFPP), site (PFPS) or area (PFA).
• The similarity of symptoms and signs caused by C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola with those of other Colletotrichum species makes impossible the detection of the pathogens based on symptoms and signs (e.g. fruiting bodies).
• The lack of rapid diagnostic methods based on serological or molecular approaches does not allow proper in planta identification of the pathogens at entry. Thorough post-entry laboratory analyses may not be feasible for certain commodities as isolation in pure culture is needed prior to proceed with DNA extraction and molecular identification based on multigene sequencing.
• The wide host range of some of those Colletotrichum species (i.e. C. siamense) limits the possibility to develop standard diagnostic protocols for all potential hosts.
• The genome plasticity and the possibility of sexual reproduction leading to genetic recombination in C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola may limit the efficacy of chemical control approaches by favouring the selection of fungicideresistant populations.

Uncertainty
Uncertainty on the actual distribution of the five Colletotrichum species in the EU, particularly with respect to records where multilocus gene sequencing analysis was not used for the identification of the isolated Colletotrichum species.

Conclusions
Of the five Colletotrichum species, C. aenigma and C. siamense are reported to be present in Italy and C. alienum in Portugal, including Madeira Islands, with a restricted distribution. C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola satisfy the criteria that are within the remit of EFSA to assess for these species to be regarded as potential Union quarantine pests (Table 10).

Supporting measure Summary
Risk element targeted (entry/establishment/ spread/impact) In the case of C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola delimitation of a buffer zone around an outbreak area can prevent spread of the pathogens and maintain a pest-free area, site or place of production.
Surveillance C. aenigma, C. alienum and C. siamense have been reported to be present in the EU. Surveillance would be an efficient supporting measure to define the actual distribution of each of those pathogens in the affected MSs and prevent its spread. analysis was not used for the identification of the isolated Colletotrichum species.

Spread
Pest potential for entry, establishment and spread in the EU (Section 3.4) C. aenigma, C. alienum and C. siamense have already entered the EU and they may further enter into, become established in, and spread within the EU. Similarly, C. perseae and C. theobromicola could potentially enter into, become established in, and spread within the EU territory.. The main pathways for the entry/further entry of the pathogens into, and spread within, the EU territory are: (i) host plants for planting, and (ii) fresh fruit of host plants, originating in infested third countries. Spores of the pathogens may be also present as contaminants on other substrates (e.g. nonhost plants, and other objects, etc.) imported into the EU, although these are considered minor pathways for the entry of the pathogens into the EU territory. C. aenigma, C. alienum and C. siamense are present in the EU, which indicates that both the biotic (host availability) and abiotic (climate suitability) factors occurring in parts of the EU are also favourable for the establishment of C. perseae and C. theobromicola, too. Following establishment, the five Colletotrichum species could spread within the EU territory by natural and human-assisted means.

None
Potential for consequences in the EU (Section 3.5) The introduction and spread of the pathogens in the EU is likely to have yield and quality impacts in some parts of the territory.
No associated crop losses have been reported so far from Italy and Portugal where C. aenigma, C. siamense (Italy) and C. alienum (Portugal) occur locally.

Available measures (Section 3.6)
Although not specifically targeted against C. aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola, existing phytosanitary measures mitigate the likelihood of the pathogens' entry into the EU territory. Potential additional measures also exist to further mitigate the risk of entry into, establishment within, or spread of the pathogens within the EU.
None A walk-in, static, closed place of crop production with a usually translucent outer shell, which allows controlled exchange of material and energy with the surroundings and prevents release of plant protection products (PPPs) into the environment. Hitchhiker An organism sheltering or transported accidentally via inanimate pathways including with machinery, shipping containers and vehicles; such organisms are also known as contaminating pests or stowaways (Toy and Newfield, 2010). Impact (of a pest) The impact of the pest on the crop output and quality and on the environment in the occupied spatial units Introduction (of a pest) The entry of a pest resulting in its establishment (FAO, 2018) Pathway Any means that allows the entry or spread of a pest (FAO, 2018) Phytosanitary measures Any legislation, regulation or official procedure having the purpose to prevent the introduction or spread of quarantine pests, or to limit the economic impact of regulated non-quarantine pests (FAO, 2018) Quarantine pest A pest of potential economic importance to the area endangered thereby and not yet present there, or present but not widely distributed and being officially controlled (FAO, 2018) Risk reduction option (RRO) A measure acting on pest introduction and/or pest spread and/or the magnitude of the biological impact of the pest should the pest be present. A RRO may become a phytosanitary measure, action or procedure according to the decision of the risk manager Spread (of a pest) Expansion of the geographical distribution of a pest within an area (FAO, 2018) Abbreviations Colletotrichum aenigma, C. alienum, C. perseae, C. siamense and C. theobromicola: Pest categorisation