key: cord-0018982-xx712e6k authors: Koutsoumanis, Kostas; Allende, Ana; Alvarez‐Ordóñez, Avelino; Bolton, Declan; Bover‐Cid, Sara; Chemaly, Marianne; Davies, Robert; De Cesare, Alessandra; Hilbert, Friederike; Lindqvist, Roland; Nauta, Maarten; Peixe, Luisa; Ru, Giuseppe; Simmons, Marion; Skandamis, Panagiotis; Suffredini, Elisabetta; Cocconcelli, Pier Sandro; Fernández Escámez, Pablo Salvador; Prieto‐Maradona, Miguel; Querol, Amparo; Sijtsma, Lolke; Suarez, Juan Evaristo; Sundh, Ingvar; Vlak, Just; Barizzone, Fulvio; Hempen, Michaela; Herman, Lieve title: Update of the list of QPS‐recommended biological agents intentionally added to food or feed as notified to EFSA 14: suitability of taxonomic units notified to EFSA until March 2021 date: 2021-07-07 journal: EFSA J DOI: 10.2903/j.efsa.2021.6689 sha: b147b1d8e642c7147cbec9c72150c0ac3e0bf287 doc_id: 18982 cord_uid: xx712e6k The qualified presumption of safety (QPS) approach was developed to provide a regularly updated generic pre‐evaluation of the safety of biological agents, intended for addition to food or feed, to support the work of EFSA's Scientific Panels. The QPS approach is based on an assessment of published data for each agent, with respect to its taxonomic identity, the body of relevant knowledge, safety concerns and occurrence of antimicrobial resistance. Safety concerns identified for a taxonomic unit (TU) are, where possible, confirmed at the species/strain or product level and reflected by ‘qualifications’. In the period covered by this statement, no new information was found that would change the status of previously recommended QPS TUs. Schizochytrium limacinum, which is a synonym for Aurantiochytrium limacinum, was added to the QPS list. Of the 78 microorganisms notified to EFSA between October 2020 and March 2021, 71 were excluded; 16 filamentous fungi, 1 Dyella spp., 1 Enterococcus faecium, 7 Escherichia coli, 1 Streptomyces spp., 1 Schizochytrium spp. and 44 TUs that had been previously evaluated. Seven TUs were evaluated: Corynebacterium stationis and Kodamaea ohmeri were re‐assessed because an update was requested for the current mandate. Anoxybacillus caldiproteolyticus, Bacillus paralicheniformis, Enterobacter hormaechei, Eremothecium ashbyi and Lactococcus garvieae were assessed for the first time. The following TUs were not recommended for QPS status: A. caldiproteolyticus due to the lack of a body of knowledge in relation to its use in the food or feed chain, E. hormaechei, L. garvieae and K. ohmeri due to their pathogenic potential, E. ashbyi and C. stationis due to a lack of body of knowledge on their occurrence in the food and feed chain and to their pathogenic potential. B. paralicheniformis was recommended for the QPS status with the qualification ‘absence of toxigenic activity’ and ‘absence of genetic information to synthesize bacitracin’. The European Food Safety Authority (EFSA) asked the Panel on Biological Hazards (BIOHAZ) to deliver a Scientific Opinion on the maintenance of the qualified presumption of safety (QPS) list. The QPS list contains biological agents, intentionally added to food and feed, which have achieved QPS status. The request included three specific tasks as mentioned in the Terms of Reference (ToR). The QPS process was developed to provide a harmonised generic pre-evaluation procedure to support safety risk assessments of biological agents performed by EFSA's scientific Panels and Units. This process assesses the taxonomic identity, body of relevant knowledge and safety of biological agents. Safety concerns identified for a taxonomic unit (TU) are, where possible, confirmed at strain or product level, reflected as 'qualifications' that should be assessed at the strain level by EFSA's Scientific Panels. A generic qualification for all QPS bacterial TUs applies in relation to the absence of acquired genes conferring resistance to clinically relevant antimicrobials (EFSA, 2008) . The list of microorganisms is maintained and re-evaluated approximately every 6 months in a Panel Statement. The Panel Statement also includes the evaluation of microbiological agents newly notified to EFSA within the previous 6-month period. The first ToR requires ongoing updates of the list of biological agents notified to EFSA, in the context of a technical dossier for safety assessment. The overall list (https://doi.org/10.5281/zenodo. 4917414) was updated with the notifications received between October 2020 and March 2021. Within this period, 78 notifications were received by EFSA, of which 55 were proposed for evaluation in feed, 13 for use as food enzymes, food additives and flavourings, 8 as novel foods and 2 as plant protection products. The new notifications received between October 2020 and March 2021 are included in the current Statement (see Appendix F). The second ToR concerns the revision of the TUs previously recommended for the QPS list and their qualifications. For this revision, articles published from July until December 2020 were assessed. The articles were retrieved and assessed through an extensive literature search (ELS) protocol available in Appendix B (see https://doi.org/10.5281/zenodo.5016562) and the search strategies in Appendix C (see https://doi.org/10.5281/zenodo.5016711). No new information was found that would affect the QPS status of those TUs or their qualifications. Schizochytrium limacinum, which is a synonym for Auranthiochytrium limacinum, is added to the QPS list. The third ToR requires a (re)assessment of new TUs notified to EFSA, for their suitability for inclusion in the updated QPS list at the Knowledge Junction in Zenodo (https://doi.org/10.5281/zenod o.4917383, Appendix E) . Seven of the 78 notifications received, corresponding to 7 TUs, were evaluated for possible QPS status; Corynebacterium stationis and Kodamaea ohmeri were re-assessed because an update was requested in relation to the current mandate. Five TUs (Anoxybacillus caldiproteolyticus, Bacillus paralicheniformis, Enterobacter hormaechei, Eremothecium ashbyi and Lactococcus garvieae) were assessed for the first time. The following conclusions were drawn: • Anoxybacillus caldiproteolyticus is not recommended for QPS status due to a lack of a body of knowledge on its occurrence in the food and feed chain. • B. paralicheniformis is recommended for the QPS status with the qualification 'absence of toxigenic activity' and 'absence of genetic information to synthesize bacitracin'. • Corynebacterium stationis is not recommended for QPS status due to a lack of a body of knowledge in relation to its occurrence in the food and feed chain and to possible safety concerns in relation to human health. • Enterobacter hormaechei is not recommended for QPS status due to its pathogenic potential for humans and animals. • Eremothecium ashbyi is not recommended for QPS status due to a lack of a body of knowledge on its occurrence in the food and feed chain and its pathogenicity for plants. • Kodamaea ohmeri is not recommended for QPS status due to safety concerns in relation to human health. • Lactococcus garvieae is not recommended for QPS status due to its pathogenic potential for humans and animals. The remaining 71 notifications were excluded from QPS evaluation for the following reasons: 25 notifications were related to microorganisms that are generally excluded from QPS evaluation (16 were notifications of filamentous fungi, 1 of Enterococcus faecium, 7 of Escherichia coli, 1 Streptomyces spp.), 2 notifications (Dyella spp. and Schizochytrium sp. strain CABIO-A-2) were only a genus, or a strain in the case of Schizochytrium, and not a species and therefore not suitable for the QPS approach and 44 (including the newly classified Lactiplantibacillus plantarum, previously known as Lactobacillus plantarum) were related to TUs that already have QPS status and did not require further evaluation. 1. The qualified presumption of safety (QPS) approach was developed by the EFSA Scientific Committee to provide a generic concept for risk assessment within the European Food Safety Authority (EFSA) for microorganisms intentionally introduced into the food chain, in support of the respective Scientific Panels and Units in the context of market authorisations for their use in food and feed, requiring an EFSA safety assessment (EFSA, 2007) . The list, first established in 2007, has been continuously revised and updated. A Panel Statement is published approximately every 6 months. These Panel Statements include the results of the assessment of relevant new papers related to the TUs with QPS status. They also contain the assessment of newly arrived TUs to the EFSA Units on Feed, Food Ingredients and Packaging (FIP), Nutrition, Pesticides and Genetically Modified Organisms (GMO). After 3 years, a QPS opinion is published summarising the results of the Panel Statements published in that period. Background and Terms of Reference as provided by EFSA A wide variety of microorganisms are intentionally added at different stages of the food and feed chain. In the context of applications for market authorisation of these biological agents used, either directly or as sources of food and feed additives, food enzymes and plant protection products, EFSA is requested to assess their safety. EFSA's work on QPS activities began in 2004 when the Scientific Committee issued a scientific opinion in continuation of the 2003 working document 'On a generic approach to the safety assessment of microorganisms used in feed/food and feed/food production' prepared by a working group consisting of members of the former Scientific Committee on Animal Nutrition, the Scientific Committee on Food and the Scientific Committee on Plants of the European Commission. 1 The document, made available for public consultation, proposed the introduction of the concept of Qualified Presumption of Safety (QPS), to be applied to selected groups of microorganisms. Microorganisms not considered suitable for QPS status would remain subject to a full safety assessment. EFSA management asked its Scientific Committee to consider whether the QPS approach could be applied to the safety assessment of microorganisms across the various EFSA Scientific Panels. In doing so, the Committee was required to take into account the response of the stakeholders to the QPS approach. In its 2005 opinion (EFSA Scientific Committee, 2005), the Scientific Committee concluded that the QPS approach could provide a generic assessment system that could be applied to all requests received by EFSA for the safety assessments of microorganisms deliberately introduced into the food and feed chain. Its introduction was intended to improve transparency and ensure consistency in the approach used across the EFSA Panels. Applications involving a taxonomic unit belonging to a species that falls within a QPS group do not require a full safety assessment. Several taxonomic units (usually species for bacteria and yeasts; families for viruses) have been included in the QPS list, either following notifications to EFSA, or proposals made initially by stakeholders during a public consultation in 2005, even if they were not yet notified to EFSA (EFSA Scientific Committee, 2005) . The EFSA Scientific Committee reviewed the range and numbers of microorganisms likely to be the subject of an EFSA Opinion and, in 2007, published a list of microorganisms recommended for the QPS list. In their 2007 opinion (EFSA, 2007) , the Scientific Committee recommended that a QPS approach should provide a generic concept to prioritise and to harmonise safety risk assessment of microorganisms intentionally introduced into the food chain, in support of the respective Scientific Panels and EFSA Units in the frame of the market authorisations for their use in the food and feed chain. The same Committee recognised that there would have to be continuing provision for reviewing and modifying the QPS list and in line with this recommendation, the EFSA Panel on Biological Hazards (BIOHAZ) took the prime responsibility for this and started reviewing annually the existing QPS list. In 2008, the first annual QPS update was published (EFSA, 2008) . In 2014, the BIOHAZ Panel, in consultation with the Scientific Committee, decided to change the revision procedure; the overall assessment of the taxonomic units previously recommended for the QPS list (EFSA BIOHAZ Panel, 2013) was no longer carried out annually but over a 3-year period. From 2017, the search and revision of the possible safety concerns linked to those taxonomic units started instead to be carried out every 6 months through extensive literature searches (ELS). The update of the 2013 QPS list (EFSA BIOHAZ Panel, 2013) was done in 2016 (EFSA BIOHAZ Panel, 2017) . From 2016 on, the QPS list (https://doi.org/10.5281/zenodo.1146566) and the list of notifications to EFSA (https://doi.org/10.5281/zenodo.3607183) are constantly updated, independent of the QPS opinion and available at the Knowledge Junction in Zenodo. The most recent QPS opinion (EFSA BIOHAZ Panel, 2020a) summarises the main results of the 3-year ELS on the QPS TUs, together with an update of the process for granting QPS status. In the meantime, every 6 months a Panel Statement, compiling the assessments for a QPS status of the microbiological agents notified to EFSA requested by the Feed Unit, the Food Ingredients and Packaging (FIP) Unit, the Nutrition Unit, the Pesticides Unit and the Genetically Modified Organisms (GMO) Unit, as well as the summary of each 6-month ELS exercise, has been produced and published. Each QPS Panel Statement contains the evaluations of the new notifications for microorganisms submitted for possible QPS status. It also contains the result of a standardized extensive literature search performed every 6 months regarding possible new safety concerns related to the TUs already included in the QPS list. The data identified are used to decide whether any TU may or may not remain on the QPS list, and whether any qualifications need to be revised. Establishing a QPS status is based on four pillars: [1] the taxonomic grouping (TU) for which QPS is sought ('taxonomic identification'); [2] whether sufficient relevant information is available about the proposed group of organisms to conclude on human/animal exposure by food/ feed ('body of knowledge'); [3] whether the grouping proposed contains known pathogens ('safety') and, finally, [4] the intended end use ('intended use'). If a hazard related to a TU is identified, which can be tested at the strain or product level, a 'qualification' to exclude that hazard may be established and added. The subject of these qualifications for the microbial strain under investigation is evaluated by the EFSA Unit to which the application dossier has been allocated. Absence of acquired genes coding for resistance to antimicrobials relevant for humans and animals is a generic qualification for all bacterial TUs; the absence of antimycotic resistance should be proven if the pertinent yeasts are to be used as viable organisms in the food or feed chains. The qualification 'for production purpose only' implies the absence of viable cells of the production organism in the final product and can also be applied to food and feed products based on microbial biomass (EFSA BIOHAZ Panel, 2020a). Because the QPS evaluation is, after its initial creation, only triggered through an application dossier notified to EFSA, the QPS list is not exhaustive. In summary, the QPS evaluation provides a generic safety pre-assessment approach for use within EFSA that covers safety concerns for humans, animals and the environment. In the QPS concept, a safety assessment of a defined taxonomic unit is performed independently of the legal framework under which the application is made in the course of an authorisation process. Although general human safety is part of the evaluation, specific issues connected to type and level of exposure of users handling the product (e.g. dermal contact, inhalation, ingestion) are not addressed. In the case of Genetically Modified Microorganisms (GMM) for which the species of the recipient strain qualifies for the QPS status, and for which the genetically modified state does not give rise to safety concerns, the QPS approach can be extended to genetically modified production strains (EFSA BIOHAZ Panel, 2018) . The assessment of potential allergenic microbial residual components is beyond the QPS remit; however, if there is science-based evidence for a microbial species it is reported. These aspects are separately assessed, where applicable, by the EFSA Panel responsible for assessing the application. The lowest TU for which the QPS status is granted is the species level for bacteria, yeasts and protists/algae, and family for viruses. Filamentous fungi, bacteriophages, Streptomycetes, Oomycetes, Enterococcus faecium, Escherichia coli and recently also Clostridium butyricum (EFSA BIOHAZ Panel, 2020a,c) are excluded from the QPS assessments based on an ambiguous taxonomic position or the possession of potentially harmful traits. The Terms of Reference are as follows: ToR 1: Keep updated the list of biological agents being notified in the context of a technical dossier to EFSA Units such as Feed, Pesticides, Food Ingredients and Packaging (FIP) and Nutrition, for intentional use directly or as sources of food and feed additives, food enzymes and plant protection products for safety assessment. ToR 2: Review taxonomic units previously recommended for the QPS list and their qualifications when new information has become available. The latter is based on a review of the updated literature aiming at verifying if any new safety concern has arisen that could require the removal of a taxonomic unit from the list, and to verify if the qualifications still efficiently exclude safety concerns. ToR 3: (Re) assess the suitability of new taxonomic units notified to EFSA for their inclusion in the QPS list. These microbiological agents are notified to EFSA and requested by the Feed Unit, the FIP Unit, the Nutrition Unit or by the Pesticides Unit. Data and methodologies In reply to ToR 3, (re)assessment of the suitability of TUs notified within the time period covered by this Statement (from October 2020 to March 2021) was carried out. The literature review considered the identification, the body of knowledge, the potential safety concerns related to human and animal health and to organisms in the environment (EFSA BIOHAZ Panel, 2020a). The environmental risk assessment of plant protection products is not included in the QPS assessment but carried out by the Pesticide Peer Review (PPR) Unit based on the information in the application dossier. The knowledge on relevant acquired antimicrobial resistance (AMR) is reflected in the safety sections of this document. Relevant databases, such as PubMed, Web of Science, CAB Abstracts or Food Science Technology Abstracts (FSTA) and Scopus, were searched. More details on the search strategy, search keys and approach are described in Appendix A. Only the literature that is considered, based on expert judgement, to be relevant for the QPS assessment is reflected in the Statement. Only valid TUs covered by the relevant international committees on the nomenclature for microorganisms are considered for the QPS assessment. In response to ToR 1, the EFSA Units were asked to update the list of biological agents being notified to EFSA. A total of 78 notifications were received between October 2020 and March 2021, of which 55 were for evaluation for use in feed, 13 for use as food enzymes, food additives and flavourings, 8 as novel foods and 2 as plant protection products (Table 1) . 2 In response to ToR 3, seven of the 78 notifications, corresponding to seven TUs, were evaluated for possible QPS status, five of these (Anoxybacillus caldiproteolyticus, Bacillus paralicheniformis, Enterobacter hormaechei, Eremothecium ashbyi and Lactococcus garvieae) for the first time. The other two, Corynebacterium stationis and Kodamaea ohmeri, were re-assessed because an update was requested in the current mandate. The remaining 71 notifications were excluded from QPS evaluation for the following reasons: 25 notifications were related to microorganisms that are generally excluded from QPS evaluation (16 were notifications of filamentous fungi, 1 of Enterococcus faecium, 7 of Escherichia coli, 1 Streptomyces spp.), two notifications were not suitable for the QPS approach, i.e. Dyella spp. because it is a genus and Schizochytrium sp. strain CABIO-A-2 because it is a strain, not a species, and 44 (including newly classified Lactiplantibacillus plantarum, previously known as Lactobacillus plantarum) were related to TUs that already had QPS status and did not require further evaluation in this mandate. 2 Eremothecium ashbyi was previously considered to be a filamentous fungus and excluded from QPS assessment. However, as a result of the recently defined distinction between yeasts and filamentous fungi (EFSA BIOHAZ Panel, 2021), it is now classified as a yeast and included in the QPS assessment. In reply to ToR 2, concerning the revision of the TUs previously recommended for the QPS list and their qualifications, an extensive literature search (ELS) was conducted as described in Appendix B -ELS protocol, see https://doi.org/10.5281/zenodo.5016562, and in Appendix C Search strategiessee https://doi.org/10.5281/zenodo.5016711, respectively. The Artificial Intelligence (AI) function was used for pre-screening of papers for Bifidobacterium spp., lactobacilli, Lactococcus lactis, Bacillus spp. and yeasts, followed by a second screening of those articles carried out by two experts. The aim of the ELS was to identify any publicly available scientific studies reporting on safety concerns for humans, animals or the environment, caused by QPS organisms since the previous QPS review (i.e. publications from July to December 2020). For case reports of human infections or intoxications, important additional information includes whether any negative impacts are confined to persons with conditions favouring opportunistic infections, for example immunosuppression, and whether transmission occurred through food or other routes, when described (e.g. medical devices). Studies indicating the presence of virulence factors (e.g. toxins and enzymes that may contribute to the pathogenicity of the microorganism) in the TU are also reported as relevant when identifying potential safety concerns. Several of the QPS-TUs are sporadically reported as causing infections in individuals with recognised predisposing conditions for the acquisition of opportunistic infections, e.g. cardiovascular conditions associated with endocarditis, people in the lower or upper age spectrum, or with other conditions which can lead to impairment of the immunological system, such as patients subjected to transplants, undergoing cancer therapy, suffering from physical trauma or tissue damage or HIV patients. Moreover, gastrointestinal tract-related conditions with mucosal impairment can also be a predisposing factor for infection. Previous use of the microorganisms being assessed as food supplements for humans was reported in many of these cases. A living microorganism used as a food supplement does not fall under the remit of the QPS assessment because regulation does not require an EFSA assessment. Nevertheless, the QPS assessment takes into consideration these reports, extracting relevant information whenever justified. For a detailed protocol of the process and search strategies, refer to Appendices B and C. After removal of duplicates, 2,770 records were submitted to the title screening step, which led to the exclusion of 2,581 of these. The remaining 189 records were found eligible for the title and abstract screening step, which led to the exclusion of 113 of these. Of the 76 articles that finally reached the article evaluation step (full text), 39 were considered to report a potential safety concern and were further analysed. The flow of records from their identification by the different search strategies (as reported in Appendix C) to their consideration as potentially relevant papers for QPS is shown in Table 2 . The search strategy (key words, literature databases, number of papers found) followed for the assessment of the suitability of TUs notified to EFSA for their inclusion in the updated QPS list (reply to ToR 3) can be found in Appendix A. C. stationis was recently evaluated (EFSA BIOHAZ Panel, 2020b) and was not recommended as QPS due to a lack of body of relevant knowledge. C. stationis (synonym Achromobacter stationis) is a valid species with standing in nomenclature. It was described by Bernard et al. (2010) , grouping Brevibacterium stationis ATCC 14403 and C. ammoniagenes ATCC 6872. Those strains can be discriminated from other Corynebacterium species by 16S rRNA gene and rpoB sequencing. The body of knowledge for C. stationis is limited. The type strain was isolated from seawater (Bernard et al., 2010) . Other strains of this species were isolated from a human infant stool sample (Bernard et al., 2010) and found in a population community from a compost produced by different types of wastewater sludge (Tashiro et al., 2016) . C. stationis is used in experimental trials as an indicator for improving UV seawater disinfection treatment (Rubio et al., 2013) . A strain of C. stationis was used to produce disodium 5 0 -inosinate for use as a feed additive (EFSA FEEDAP Panel, 2020). Two clinical C. stationis isolates were obtained from blood cultures from a 62-year-old male with a chest infection, and a 66-year-old female; no further clinical information was provided (Bernard et al., 2010) . In a recent article, C. stationis was identified among bacteria causing diabetic foot infection and presenting resistance to antibiotics (Henciya et al., 2020) . C. stationis has been isolated from the raw milk of cows with mastitis ( Conclusion on a recommendation for QPS status C. stationis cannot be recommended for QPS status due to a lack of body of knowledge on its occurrence in the food and feed chain and possible safety concerns in relation to human and animal health. Kodamaea ohmeri was recently evaluated (EFSA BIOHAZ Panel, 2020b) and not recommended for QPS due to safety concerns. K. ohmeri is a yeast within the Saccharomycetales order. A synonym of K. ohmeri is Pichia ohmeri and the teleomorphic form is Candida guilliermondii var. membranaefaciens (Kurtzman et al., 2011) . It is a biologically under-investigated taxon and its relationship with other taxa within the Order Saccharomycetales is uncertain (Lumbsch and Huhndorf, 2007) . Currently, it classified in the Class Ascomyceta and the Saccharomycetaceae family. Correct identification apparently remains a challenge. Body of knowledge K. ohmeri has been recovered from a broad variety of sources. From a biotechnological point of view, this species is used in the production of xylitol from glucose and the conversion of xylulose to xylitol (Kurtzman et al., 2011) . Up to 2020, more than 70 cases of invasive fungaemia due to the yeast K. ohmeri have been reported (Kanno et al., 2017; Ioannou and Papakitsou, 2020) . A patient with a severe fungaemic pulmonary infection was diagnosed with K. ohmeri (Kanno et al., 2017) . The above information confirms our previous Statement (EFSA BIOHAZ Panel, 2020b) , that K. ohmeri cannot be recommended for QPS status due to safety concerns in relation to human health. Taxonomic units to be evaluated for the first time 3.2.1. Bacteria Identity A. caldiproteolyticus is an aerobic, Gram-positive, sporulating, thermophilic bacterial species with standing in nomenclature (Coorevits et al., 2012) . The type strain was isolated from sewage and initially classified as Geobacillus caldoproteolyticus (Chen et al., 2004) . A. caldiproteolyticus was first isolated and used for the production of a thermostable protease (Chen et al., 2004) , a phenotypic trait later confirmed by another strain that generated a keratinase (Reis et al., 2020) . Strains of the species have also been isolated from hot springs (Najar et al., 2018) . A. caldiproteolyticus has been implicated in the generation of biofilms on dairy abiotic surfaces (Karaca et al., 2019) . No safety concerns were reported for A. caldiproteolyticus. A. caldiproteolyticus cannot be recommended for the QPS status due to lack of body of knowledge on its occurrence in the food and feed chain. Identity B. paralicheniformis is a valid species name with standing in nomenclature (Oren and Garrity, 2016) . B. paralicheniformis are Gram-positive, facultatively anaerobic, mobile, endospore-forming rods; producing catalase but not oxidase. Based on phylogenomic analysis of all available genomes of the B. licheniformis group, a number of strains, originally designated as B. licheniformis, were reclassified as a new species; B. paralicheniformis (Dunlap et al., 2015) . The type strain was isolated from cheonggukjang, a Korean fermented soybean food product. B. paralicheniformis has been isolated from the food product cheonggukjang (Dunlap et al., 2015) , from kimchi, a traditionally, spontaneously fermented vegetable Korean food product (Khan et al., 2018) and from whey powder as a contaminant (McHugh et al., 2018) . B. paralicheniformis were reported as plant-associated bacteria, displaying growth promoting abilities (Annapurna et al., 2018) and the potential to be used as phytopathogen biocontrol agents Pylro et al., 2019; Jinal et al., 2020; Ramirez-Carino et al., 2020) . B. paralicheniformis strains also have the potential to be used for biotechnological purposes and for bioremediation (Rahman et al., 2019; Khan et al., 2018; SanthaKalaikumari et al., 2021; Ganesh Kumar et al., 2021) . Ahire et al. (2020) reported the production of a bacitracin, an antimicrobial peptide active against a range of Gram-positive bacteria, by strains of B. paralicheniformis. The ability to produce bacitracin is reported for a subset of B. paralicheniformis (Du et al., 2019). No negative effects in relation to B. paralicheniformis were reported. All strains classified as B. paralicheniformis are reported to have phenotypic resistance to erythromycin as well as chromosomal erythromycin resistance genes (Lee and Jeong, 2017; Agersø et al., 2018 Agersø et al., , 2019 Jeong et al., 2020) . B. paralicheniformis is recommended for the QPS status with the qualification 'absence of toxigenic activity' and 'absence of genetic information to synthesize bacitracin'. Enterobacter hormaechei Identity E. hormaechei is a TU with standing in nomenclature. It was first described by O'Hara et al. (1989) amended by Hoffmann et al. (2005) and updated by Oren and Garrity (2017) . It is currently placed within the E. cloacae complex, together with five other species and six additional genovars (Hoffmann et al., 2005; Sutton et al., 2018) , but further taxonomic restructuration might occur based on the description of new species and subspecies by Wu et al. (2020) . E. hormaechei has been isolated as a contaminant from food (Indugu et al., 2020; Sadek et al., 2020) , as an endophyte from plants (Khalaf and Raizada, 2020; Tshishonga and Serepa-Dlamini, 2019) and from the gastrointestinal tracts of insects (Asimakis et al., 2019) . Isolates of E. hormaechei are able to degrade lutein, a plant carotenoid (Zhong et al., 2017) . Enhancing soil fertility and P-and Kuptake by plants have been reported for E. hormaechei (Roslan et al., 2020) . Safety concerns E. hormaechei is an opportunistic human pathogen, causing mainly nosocomial infections (Townsend et al., 2008; Roberts et al., 2020) . It has frequently been reported as the cause of human disease in susceptible individuals, in association with multidrug resistance, including to critical antimicrobials located on mobile genetic elements (Yang et al., 2018; Yuan et al., 2019; Gou et al., 2020; Martins et al., 2020; Soliman et al., 2020) . The virulence features of E. hormaechei are insufficiently studied. The genomic plasticity of this species may explain its ability to spread in hospital environments (Paauw et al., 2009; Roberts et al., 2020) . E. hormaechei has also been associated with respiratory disease in un-weaned calves . Multiresistant E. hormaechei has been isolated from several food products. Examples are a shrimp sample from a farmers' market in the US harbouring sul1, sul2, qnrA1, oqxAB, dfrA23, blaACT, floR, fosA, tet(A), aph(6)-Id and aph(30)-Ib antibiotic resistance genes and several plasmids (Indugu et al., 2020) ; an isolate from a raw beef patty with bla VIM-1 , mcr-9 aac(6 0 )-Il, ΔaadA22, aac(6 0 )-Ib-cr, sul1, dfrA1 on a plasmid (Sadek et al., 2020) . Enterobacter hormaechei cannot be recommended for the QPS status due to its pathogenic potential for humans and animals. Identity L. garvieae is a bacterial species name with standing in nomenclature (Collins et al., 1983; Schleifer et al., 1985) . This bacterial species has been isolated from a variety of terrestrial and aquatic environments. L. garvieae has been found in raw milk and sporadically in the bacterial communities of fermented dairy products. In a single study, L. garvieae has been used as a cheese starter culture (Guarcello et al., 2016) . The pathogenicity of L. garvieae has been reviewed by Gibello et al. (2016) . This species is a recognised fish pathogen responsible for high mortality haemorrhagic septicaemia in wild fish and in fish farmed in fresh and sea water (Shahi and Mallik, 2020) . It has also been associated with bovine mastitis (Sorge et al., 2021) . L. garvieae is also a rare human pathogen of increasing clinical significance, facilitated by predisposing factors, being responsible for infections such as endocarditis, urinary tract infections and septicaemia (Gibello et al., 2016; Choksi and Dadani, 2017; Lee et al., 2020) . Conclusions on a recommendation for the QPS status L. garvieae is not recommended for the QPS status due to its pathogenic potential for humans and animals. Eremothecium ashbyi The genus Eremothecium contains five species of ascomycetous yeasts or yeast-like fungi and belongs to the family Saccharomycetaceae (Prillinger et al., 1997; Kurtzman et al., 2011) . The species E. ashbyi was first described in 1935 and it is a legitimate species and name (Mycobank, Index Fungorum). A synonym name is Crebrothecium ashbyi, which however has been little used. The species designation has repeatedly been spelled ashbyii; however, this is considered by Kurtzman et al. (2011) as an orthographic error. A special feature of E. ashbyi (and other species of the genus) is growth predominantly as pseudohyphae and absence of budding. The ascospores in this genus are sickle-or needle-shaped. Very little information is available on the ecology of E. ashbyi. It has been isolated mostly in cropping systems of tropical or subtropical regions, e.g. cotton and citrus fruit. Batra (1973) considered it to be a relatively rare fungus. No other information was found on the possible occurrence of the species in the food and feed chains. Along with other species of the genus, E. ashbyi has been widely used in biotechnology since the 1950s for production of useful compounds. The main interest has been the overproduction of riboflavins by some strains (Kapralek, 1962; Semenova et al., 2017) . Additionally, some studies report extraction of accumulated lipids and essential oils from the biomass of E. ashbyi (He et al., 2019; Vijayalakshmi et al., 2003) . There are no reports indicating that E. ashbyi might be pathogenic to humans or any other animals. However, some studies characterise it as a plant pathogen, since it has been shown to be able to cause stigmatomycosis (or 'yeast spot') in citrus fruit (Batra, 1973) , soybean (Kimura et al., 2008a) and azuki bean (Kimura et al., 2009) (Kimura et al., 2008b) . E. ashbyi is not recommended for the QPS list due to its plant pathogenicity and lack of body of knowledge on its occurrence in the food and feed chains. The summaries of the evaluation of the possible safety concerns for humans, animals or the environment described and published since the previous ELS exercise (i.e. articles published between July 2020 and December 2020 as described in Appendices B and C with reference to the articles selected as potentially relevant for the QPS exercise (Appendix D) for each of the TUs or groups of TUs that are part of the QPS list (Appendix E), are presented below. Gram-positive non-sporulating bacteria 3.3.1. Bifidobacterium spp. A search for papers potentially relevant for QPS-listed Bifidobacterium spp. provided 315 references. The artificial intelligence (AI) analysis left 150. Title screening left 14 references for abstract inspection, then 8 for a full article appraisal. This last step identified two articles dealing with safety concerns. The first article described a case of bacteraemia in a premature baby girl who was given a probiotic mixture containing, amongst other organisms, Bifidobacterium longum. The isolated strain from blood culture was compared to the one from the probiotic and was indistinguishable, but the methods of isolation and genotyping were not provided. The authors concluded that the infection was a likely adverse event of the probiotic supplementation. In the second article , the authors described a case of an old woman hospitalised with an infection in the peribronchial connective tissue caused by Bifidobacterium longum and Veillonella species, cultured from a transbronchial needle aspirate. However, the role of B. longum in the infection was not elucidated since it was identified together with Veillonella species. Based on the available evidence, the QPS status of the QPS-listed Bifidobacterium spp. is not changed. A search for potentially relevant papers on C. divergens provided eight references. No article was considered relevant at the level of title screening for this TU. Consequently, the QPS status of C. divergens is not changed. A search for papers potentially relevant to the QPS evaluation of C. glutamicum provided 34 references. One paper reached the level of title and abstract screening but did not reach full text evaluation. Therefore, no new safety concerns were identified and the QPS status of C. glutamicum is not changed. Analysis of papers referring to any of the QPS species, formerly belonging to the genus Lactobacillus and recently split into 13 new genera, provided 853 references. The AI analysis left 423 articles. Title screening of these provided 14 references for abstract inspection, which further reduced their number to 5. However, full paper review excluded all of them, either because no reliable microorganism identification procedures were described or due to uncertainty on the aetiology of the cases described. Based on the available evidence as described above, the QPS status of any of the QPS species included in the genus Lactobacillus is not changed. A search for papers potentially relevant for the QPS status of L. lactis provided 275 references. The AI analysis left 126 papers. Title and abstract screenings of these reduced their number to 11. Two papers reached full article evaluation but neither of them dealt with a possible safety concern pertaining to L. lactis. Based on the available evidence as described above, the QPS status of L. lactis is not changed. A search for papers potentially relevant for the QPS evaluation of Leuconostoc species provided 95 references. The analysis of their titles left 10 articles for title/abstract screening. Six articles, of which one was not in English, reached full text evaluation, and five dealt with possible safety concerns. These were excluded because either the isolation procedure or the identification procedures were considered unreliable. Consequently, the status of QPS-listed Leuconostoc spp. is not changed. A search for papers potentially relevant for the QPS evaluation of Microbacterium imperiale provided one reference. The analysis of the title led to this reference being discarded. Consequently, the QPS status of M. imperiale is not changed. A search for papers potentially relevant for the QPS evaluation of Oenococcus oeni provided 36 references. The analysis of their titles left two articles for title/abstract screening. One article reached full text evaluation but did not raise any safety concerns. Consequently, the QPS status of O. oeni is not changed. A search for papers potentially relevant for the QPS evaluation of Pediococcus spp. provided 193 references. The analysis of their titles left four articles for the title/abstract phase. Two articles reached the full text evaluation stage but neither of them identified a safety concern. Consequently, the status of QPS-listed Pediococcus spp. is not changed. A search for papers potentially relevant for the QPS evaluation of Propionibacterium spp. provided 36 references. Following the analysis of their titles, no articles were selected for abstract screening or the full article evaluation phase; thus, no new safety concerns were identified. Consequently, the status of QPS-listed Propionibacterium spp. is not changed. A search for papers potentially relevant for the QPS evaluation of Streptococcus thermophilus provided 79 references. The analysis of their titles left one article for title and abstract screening that did not deal with safety concerns. Therefore, no article reached the evaluation phase, and the QPS status of S. thermophilus is not changed. Bacillus spp. A search for papers potentially relevant for Bacillus spp. provided 1,123 references. The AI analysis left 557 articles. The analysis of their titles by two experts left 13 articles for the abstract phase and, from these, four articles passed to the full text phase for further analysis. Two papers did not deal with safety concerns. Two papers were further analysed. described Bacillus licheniformis as possibly involved in bovine mastitis. The paper had methodological flaws in relation to strain identification and did not document source attribution. Princess et al. (2020) described the isolation of B. clausii from the blood of a patient, who had been treated with a B. clausii probiotic. The identification via the blood isolate was performed by MALDI-TOF and the link to the probiotic strain was not confirmed. The authors reported the patient as immunocompetent, but the patient had underlying diseases, being a type II diabetic who had undergone several surgical procedures and was being treated for a tracheal infection while taking the probiotic. The ELS did not identify any information that would change the status of members of Bacillus spp. included in the QPS list. A search for papers potentially relevant for G. stearothermophilus provided 1,123 references. The AI analysis left 557 articles. The analysis of their titles by two experts left 13 articles and for four of these the full text was analysed. None dealt with this species. Consequently, the QPS status G. stearothermophilus is not changed. A search for papers potentially relevant for the QPS evaluation of P. nishizawae provided one reference, which was excluded during title screening. Consequently, the QPS status of P. nishizawae is not changed. A search for papers potentially relevant to the QPS evaluation of G. oxidans and Xanthomonas campestris provided 200 references. The analysis of their titles left two articles, but these were excluded following the title and abstract screening; therefore, no paper reached the final selection phase for this TU. Consequently, the QPS status of G. oxydans is not changed. The search for papers potentially relevant for the QPS evaluation of Gluconobacter oxidans and X. campestris provided 200 references. The analysis of their titles left two articles, which reached the evaluation phase for this TU, but neither eventually dealt with health nor safety concerns. Consequently, the QPS status of X. campestris is not changed. The ELS searches for potentially relevant studies on the yeasts with QPS status provided 1,349 references. The AI analysis left 673 articles. After title screening by two experts, 74 studies remained for the title/abstract phase, and from these 36 articles passed to the full article appraisal. Out of these, 21 reported a possible safety concern. Two studies were considered relevant for the QPS evaluation but did not directly report safety concerns. Boontham et al. (2020) proposed a name change of the QPS species Candida cylindracea to Limtongozyma cylindracea. Libkind et al. (2020) discussed the use of full genome sequencing for taxonomic identification of yeasts. Twenty-one studies discussed potentially relevant safety concerns for QPS yeast species, which are discussed below. For the species Candida cylindracea, Kluyveromyces lactis, Komagataella pastoris, Komagataella phaffi, Ogataea angusta, Saccharomyces bayanus, Saccharomyces pastorianus, Schizosaccharomyces pombe, Xanthophyllomyces dendrorhous and Zygosaccharomyces rouxii, no safety concerns were newly reported. Consequently, the QPS status does not change for these species. The anamorph name of C. jadinii is Candida utilis. In a retrospective study for 1999-2018, identified 7927 yeast isolates (out of 602,963 isolates in total) from patients with suspected sepsis in intensive care units in a hospital in India. They reported that C. jadinii represented 2.8% of the yeasts and 0.03% of total isolates (identification by conventional methods and MALDI-TOF MS, but no confirmation with DNA-based molecular methods). found in a retrospective study (2001) (2002) (2003) (2004) (2005) (2006) (2007) (2008) (2009) (2010) (2011) (2012) (2013) (2014) (2015) (2016) (2017) (2018) on isolates from patients with candidaemia in a hospital in Malaysia, that a very low incidence (one isolate or 0.1%) was C. jadinii. In a retrospective study of isolates from patients with fungaemia in 12 hospitals in Italy , one of the isolates (0.3%) was C. jadinii (identification by conventional methods and MALDI-TOF MS). The incidence was very low and identification by MALDI-TOF was not further confirmed by a DNA-based molecular methodology. The inconsistency of the identification, together with the scarcity of linkage of this organism to pathology, resulted in the decision that the QPS status of C. jadinii is not changed. The anamorph name of H. uvarum is Kloeckera apiculata. investigated the causes of cranberry fruit rot (CFR) in a cranberry farm in Wisconsin, USA. They show that in rare cases (CFR is usually caused by filamentous fungi), H. uvarum can cause CFR and thus cause spoilage of harvested cranberries. This new report did not add any new information that would change the current QPS status of this species. The anamorph name of K. marxianus is Candida kefyr. reported bloodstream infection with K. marxianus in a 61-year-old male. He was immunocompromised due to acute myeloid leukaemia and chemotherapy and prophylaxic antibiotic treatment, and species identification (growth on chromogenic media and MALDI-TOF MS) did not apply DNA-based molecular methods. A study in a hospital in India reported that 2% of yeasts recovered from the saliva of patients with oral squamous cell carcinoma were K. marxianus. The patients, however, had not been diagnosed with fungal infection. found in a retrospective study (2001) (2002) (2003) (2004) (2005) (2006) (2007) (2008) (2009) (2010) (2011) (2012) (2013) (2014) (2015) (2016) (2017) (2018) on isolates from patients with candidaemia in a hospital in Malaysia, that one of the isolates (incidence only 0.1%) was K. marxianus. In a retrospective study of isolates from patients with fungaemia in 12 hospitals in Italy , one of the isolates (low incidence of 0.3%) was K. marxianus (identification by conventional methods and MALDI-TOF MS). Eghtedar identified yeast isolates from HIV patients in a hospital in Iran and a non-HIV control group. One isolate in each group (corresponding to only 1% of isolates) was K. marxianus. A study of isolates from women in Iran diagnosed with vulvovaginal infection showed that K. marxianus may in rare cases (prevalence 1.6%) cause vulvovaginal infections. The results of two additional studies reporting a low prevalence (< 2%) of K. marxianus in vulvovaginal infections (Ignjatovi c et al., 2020) and nosocomial fungal infections in patients that had undergone abdominal surgery could not be appropriately evaluated since there were uncertainties in the species identification. In conclusion, the literature update mentioned mainly the isolation of K. marxianus from patients who are immunocompromised and/or have underlying disease and there is a scarcity of linkage of this organism to pathology. The prevalence of K. marxianus in the retrospective studies was very low. Also, methodological problems concerning identification (no confirmation by use of DNA-based molecular methods) and source attribution were noted. Thus, the papers did not identify any information that would change the QPS status of K. marxianus. The anamorph name of Y. lipolytica is Candida lipolytica. Chi et al. (2021) report a case where a 44-year-old man in Taiwan treated for gastric adenocarcinoma by gastric surgery and chemotherapy developed fungaemia caused by Y. lipolytica. Predisposing conditions were a central venous catheter and an infection with Acinetobacter baumannii. found in a retrospective study (2001) (2002) (2003) (2004) (2005) (2006) (2007) (2008) (2009) (2010) (2011) (2012) (2013) (2014) (2015) (2016) (2017) (2018) on isolates from patients with candidaemia in a hospital in Malaysia, that four of the isolates (incidence only 0.3%) were Y. lipolytica. In a retrospective taxonomic study on 85 'uncommon Candida species' isolates from patients with candidaemia in hospitals in Taiwan , three isolates were identified as Y. lipolytica by conventional methods, MALDI-TOF MS and DNA sequencing. However, no information was given on pathology or any predisposing factors for infection. also showed that conventional methods often led to misidentification, especially of Candida sake and the QPS species D. hansenii. The literature update did not identify any information that would change the current QPS status of Y. lipolytica. The anamorph name of D. hansenii is Candida famata. Six references related to possible concerns for human safety were identified. reported the prevalence of Candida species in the saliva of patients with oral squamous cell carcinoma, other oral potentially malignant disorders and healthy cohorts in a hospital in India and 17% were identified as C. famata. found in a retrospective study (2001) (2002) (2003) (2004) (2005) (2006) (2007) (2008) (2009) (2010) (2011) (2012) (2013) (2014) (2015) (2016) (2017) (2018) on isolates from patients with candidaemia in a hospital in Malaysia, that 1.7% of the isolates were C. famata from patients with oral squamous cell carcinoma. All these patients had disorders that could predispose them to infection. In a retrospective taxonomic study on 85 'uncommon Candida species' isolates from patients with candidaemia in hospitals in Taiwan , four isolates were identified as C. famata by conventional methods, MALDI-TOF MS and DNA sequencing. also showed that conventional methods often led to misidentification, especially of Candida sake and the QPS species D. hansenii. In this study, no information was given on pathology or any predisposing factors for infection. report the cases of candidaemia in paediatric patients with malignancies in hospitals in Greece, and 15.8% of the isolates were described as C. famata. report five isolates of C. famata from a total of 79 microbial isolates from patients with nosocomial, opportunistic infections in a paediatric intensive care unit at a hospital in Saudi Arabia. The report of also has problems with the identification methodology used; the yeast isolates were identified only by using conventional growth-based tests and not confirmed by molecular approaches. This study indicates that C famata might contribute to dermatomycosis or onychomycosis in humans. In these three reports, species identification is unclear. The retrospective studies did not provide further information on the cases reported. The reports on D. hansenii did not add any new information that would change the current QPS status of this species. The anamorph form of S. cerevisiae is not described. A synonym of this species is Saccharomyces boulardii. Four references reported safety concerns for humans and, in all of them, the identification is uncertain. report that two cases of critically ill patients who had to be hospitalised in an ICU due to Sars-CoV-2 infection received S. cerevisiae supplementation because of diarrhoea and subsequently developed a S. cerevisiae bloodstream infection. The identification was done using a morphological approach and API ID 32C. In a retrospective study of isolates from patients with fungaemia in 12 hospitals in Italy , three of the isolates (0.3%) were S. cerevisiae The reports on S. cerevisiae did not add any new information that would change the current QPS status of this species. The anamorph name of W. anomalus is Candida pelliculosa. , in a retrospective study, from a total of 602,963 blood samples from patients with suspected sepsis, identified in intensive care units in a hospital in India, that W. anomalus constituted 13.1% of yeast isolates (identification by conventional methods and MALDI-TOF MS). In a retrospective taxonomic study on 85 'uncommon Candida species' isolated from patients with candidaemia in hospitals in Taiwan , 15 isolates were identified as W. anomalus by conventional methods and MALDI-TOF MS. report the identification of 119 yeast isolates from patients diagnosed with blood-stream candidiasis in an intensive care unit of a hospital in India. Using MALDI-TOF MS, 15 isolates were identified as W. anomalus. In these three reports, species identification is uncertain because of the low resolution of the methods used for yeast identification. reported the prevalence of Candida species in the saliva of patients with oral squamous cell carcinoma, oral potentially malignant disorders and healthy cohorts in a hospital in India, and 33% were identified as C. pelliculosa in patients with oral squamous cell carcinoma. found in a retrospective study (2001) (2002) (2003) (2004) (2005) (2006) (2007) (2008) (2009) (2010) (2011) (2012) (2013) (2014) (2015) (2016) (2017) (2018) on isolates from patients with candidaemia in a hospital in Malaysia, that four of 1,175 isolates (0.3%) C. pelliculosa. The retrospective studies did not provide further information on the cases reported. Finally, Dutra et al. (2020) report a case of infection by W. anomalus in a 19-month-old Brazilian female who had a previous history of prematurity, cardiac surgery due to patent ductus arteriosus and a long period of hospital stay. The reports assessed in this period showed inconsistency in the identification, predisposing factors for human infection and/or a lack of information on the cases reported. The literature update did not identify any information that would change the current QPS status of W. anomalus. ELS was performed for all three species together, as indicated below. A search for papers potentially relevant for the QPS evaluation of protists/algae provided 84 references. The analysis of their titles left 42 articles for title/abstract screening. Eight articles reached full text evaluation. Four of eight articles dealt with potential safety concerns of which two were on A. limacinum, one on Schizochytrium limacinum and one on T. chuii (see below). A. limacinum is a member of the Thraustochytriacea and is a protist. The taxonomic classification of the genus Schizochytrium has been subject to discussion in 2007 (Yokoyama and Honda, 2007) . Based on genetic and phenotypic analysis, the authors proposed changes in the classification. The genus Schizochytrium was amended and new genera such as Aurantiochytrium and Oblongichytrium were defined. The species Schizochytrium llmacinum can also be referred to as Aurantiochytrium limacinum. No article indicated a safety concern; therefore, the current QPS status of A. limacinum is not changed. No article dealt with potential safety concerns of E. gracilis. Therefore, the current QPS status of E. gracilis is not changed. One article on T. chuii dealt with a potential safety concern, however, this concern was not confirmed after full text evaluation. Therefore, the current QPS status of T. chuii is not changed. A search for papers potentially relevant for the QPS evaluation of viruses of the Alphaflexiviridae and Potyviridae provided 31 references. After title screening, no paper reached the title/abstract screening stage; thus, no new safety concern was identified. Therefore, the current QPS status remains unchanged. A search for papers potentially relevant for the QPS evaluation of Baculoviridae provided 43 references. One article dealing with Baculoviridae passed the title screening but did not reach the full article evaluation stage, thus no new safety concern was identified. Therefore, the current QPS status remains unchanged. ToR 1: Keep updated the list of biological agents being notified, in the context of a technical dossier to EFSA Units (such as Feed, Food Ingredients and Packaging, Nutrition, Pesticides, Genetically Modified Microorganisms), for intentional use in feed and/or food or as sources of food and feed additives, enzymes, plant protection products for safety assessment: • Between October 2020 and March 2021, the list of notifications was updated with 78 notifications that were received by EFSA, of which 55 were proposed for evaluation as feed additives, 13 for use as food enzymes, food additives and flavourings, 8 as novel foods and 2 as plant protection products. ToR 2: Review taxonomic units previously recommended for the QPS list and their qualifications when new information has become available: In relation to the results of the monitoring of possible new safety concerns relevant for the QPS list, there were no results that would justify removal of any TUs from the QPS list. Schizochytrium limacinum, which is a synonym for Aurantiochytrium limacinum, is added to the QPS list. ToR 3: (Re)assess the suitability of taxonomic units notified to EFSA not present in the current QPS list for their inclusion in that list: • Out of the 78 notifications received, 44 were related to TUs that already had QPS status and did not require further evaluation. • Of the remaining 34 notifications, 27 were related to TUs excluded from QPS evaluation: 16 were notifications of filamentous fungi, 1 of Enterococcus faecium, 7 of Escherichia coli, 1 of Streptomyces spp. and 2 notifications (Dyella spp. and Schizochytrium sp. strain CABIO-A-2) were only a genus, or a specific strain, and not a species and were therefore not eligible for QPS evaluation. • Seven notifications, corresponding to 7 TUs, were evaluated for possible QPS status. Corynebacterium stationis and Kodamaea ohmeri were re-assessed because an update was requested in relation to the current mandate. Five TUs (Anoxybacillus caldiproteolyticus, Bacillus paralicheniformis, Enterobacter hormaechei, Eremothecium ashbyi and Lactococcus garvieae) were assessed for the first time. • Corynebacterium stationis is not recommended for QPS status due to a lack of body of knowledge in relation to its occurrence in the food and feed chain and to possible safety concerns in relation to human health. • Kodamaea ohmeri is not recommended for QPS status due to safety concerns in relation to human health. • Anoxybacillus caldiproteolyticus is not recommended for QPS status due to a lack of body of knowledge on its occurrence in the food and feed chain. • Bacillus paralicheniformis is recommended for the QPS status with the qualifications 'absence of toxigenic activity' and 'absence of genetic information to synthesize bacitracin'. • Enterobacter hormaechei is not recommended for QPS status due to its pathogenic potential for humans and animals. • Eremothecium ashbyi is not recommended for QPS status due to a lack of body of knowledge on its occurrence in the food and feed chain and its plant pathogenicity. • Lactococcus garvieae is not recommended for QPS status due to its pathogenic potential for humans and animals. Valid name of a fungus based on the asexual reproductive state (morphologically). Antimicrobial compounds Antibiotics, bacteriocins and/or small peptides with antimicrobial activity. Basonym name the earliest validly published name of a taxon. Synonymous name/Homotypic synonym have the same type (specimen) and the same taxonomic rank. Valid name of a fungus based on the sexual reproductive state (morphologically). Antimicrobial susceptibility testing and tentative epidemiological cutoff values for five Bacillus species relevant for use as animal feed additives or for plant protection Putative antibiotic resistance genes present in extant Bacillus licheniformis and Bacillus paralicheniformis strains are probably intrinsic and part of the ancient resistome Identification and characterization of antimicrobial peptide produced by indigenously isolated Bacillus Paralicheniformis Ubbli30 strain Pattern and frequency of hospital acquired infections in pediatric intensive care unit at Abha maternity and children hospital, Saudi Arabia Invasive potential of bacterial isolates associated with subclinical bovine mastitis Whole genome shotgun sequence of Bacillus Paralicheniformis Strain Kms 80, a Rhizobacterial Endophyte isolated from Rice Draft genome sequence of Enterobacter Hormaechei Ent5, a Component of the Symbiotic Community of Tephritid Flies Nematosporaceae (Hemiascomycetidae): Taxonomy, Pathogenicity, Distribution, and Vector Relations Assignment of Brevibacterium Stationis (Zobell and Upham 1944) Breed 1953 to the Genus Corynebacterium, as Corynebacterium Stationis Comb. Nov., and Emended Description of the Genus Corynebacterium to Include Isolates That Can Alkalinize Citrate Limtongozyma Siamensis Gen. Nov., Sp. Nov., a Yeast Species in the Saccharomycetales and Reassignment of Candida Cylindracea to the Genus Limtongozyma Thermoactive extracellular proteases of Geobacillus Caldoproteolyticus Update of the List of QPS-Recommended Biological Agents Intentionally Added to Food or Feed as Notified to EFSA 12: suitability of Taxonomic Units Notified to EFSA until Statement on the update of the list of QPSrecommended biological agents intentionally added to food or feed as notified to EFSA 13: suitability of taxonomic units notified to EFSA until EFSA Panel on Additives and Products or Substances usedin Animal Feed) Safety and Efficacy of Imp (Disodium 5 0 -Inosinate) Produced by Fermentation with Corynebacterium Stationis Kccm 80161 for All Animal Species Opinion of the Scientific Committee on a Request from Efsa Related to a Generic Approach to the Safety Assessment by Efsa of Microorganisms Used in Food/Feed and the Production of Food/ Feed Additives Molecular identification of Candida isolates by real-time PCR-high-resolution melting analysis and investigation of the genetic diversity of Candida species Low Prevalence of Antifungal Resistant Candida Africana, in the C Albicans Complex Causing Vulvovaginal Candidiasis Biodegradation of polystyrene by deep-sea Bacillus Paralicheniformis G1 and genome analysis The Zoonotic Potential of Lactococcus Garvieae: an overview on microbiology, epidemiology, virulence factors and relationship with its presence in foods Carbapenem-resistant enterobacter hormaechei St1103 with Imp-26 Carbapenemase and Esbl Gene Bla Shv-178 A large factory-scale application of selected autochthonous lactic acid bacteria for PDO Pecorino Siciliano cheese production Components and antibacterial activity of a novel essential oil from the Nutrient Broth of Eremothecium Ashbyii H4565 Antibacterial activity of halophilic bacteria against drug-resistant microbes associated with diabetic foot infections Three New Subspecies of Clinical Importance High rates of misidentification of uncommon Candida species causing bloodstream infections using conventional phenotypic methods Recurrent vulvovaginal candidosis and cluster analysis of clinical signs and symptoms: a laboratory-based investigation Whole-genome sequence analysis of multidrug-resistant enterobacter Hormaechei isolated from imported retail shrimp. Microbiol Resour Announc Kodamaea Ohmeri infections in humans: a systematic review Two genes involved in clindamycin resistance of Bacillus licheniformis and Bacillus paralicheniformis identified by comparative genomic analysis Characterisation of antagonistic Bacillus Paralicheniformis (Strain Eal) by Lc-Ms, antimicrobial peptide genes, and Isr determinants Catheter-related bloodstream infection caused by Kodamaea Ohmeri: a case report and literature review The physiology of riboflavin production by Eremothecium Ashbyi Anoxybacillus and Geobacillus Biofilms in the dairy industry: effects of surface material, incubation temperature and milk type Candidaemia in a tertiary care centre of developing country: monitoring possible change in spectrum of agents and antifungal susceptibility Draft Genome Sequences of Acinetobacter Sp. Strain Ekm10a, Enterobacter Hormaechei Ekm10e, and Enterobacter Hormaechei Ekm11e (Phylum Proteobacteria) Colonizing the Seed Surface Biogel of Echinocystis Lobata (Wild Cucumber). Microbiol Resour Announc Immobilization of an Alkaline Endopolygalacturonase purified from Bacillus Paralicheniformis Exhibits Bioscouring of Cotton Fabrics Risk prediction for candidemia in surgical intensive care unit patients Mode of transmission and morphological structures of two Eremothecium Species between Riptortus Pedestris and Soybean Eremothecium Ashbyi Causes soybean yeast-spot and is associated with Stink Bug, Riptortus clavatus Eremothecium Coryli and E. Ashbyi cause yeast spot of Azuki Bean Complete genome sequence of Bacillus paralicheniformis 14DA11, exhibiting resistance to clindamycin and erythromycin Molecular detection and sensitivity to antibiotics and bacteriocins of pathogens isolated from bovine mastitis in family dairy herds of central Mexico Towards yeast Taxogenomics: lessons from novel species descriptions based on complete genome sequences USA: The Field Museum, Department of Botany Typeability of Maldi-Tof assay for identification of Non-Aureus Staphylococci associated with bovine intramammary infections and teat apex colonization Genome and plasmid context of two RMTG-carrying enterobacter hormaechei isolated from urinary tract infections in Brazil Mesophilic sporeformers identified in whey powder by using shotgun Metagenomic sequencing. Applied and Environment Microbiology Virulence factors and antifungal susceptibility in Candida species isolated from dermatomycosis patients Rare invasive yeast infections in greek neonates and children, a retrospective 12-year study Enterobacter Hormaechei, a new species of the family Enterobacteriaceae Formerly Known as Enteric Group 75 Notification that new names of prokaryotes, new combinations and new taxonomic opinions have Notification of changes in taxonomic opinion previously published outside the Ijsem Identification of resistance and virulence factors in an epidemic Enterobacter Hormaechei Outbreak Strain Necessity to identify Candida species accurately with minimum inhibitory concentration determination in each case of bloodstream infections Does probiotic bacteremia in premature infants impact clinically relevant outcomes? A case report and updated review of literature Evolution of Fungemia in an Italian Region Phytopathogenic Filamentous (Ashbya, Eremothecium) and Dimorphic Fungi (Holleya, Nematospora) with Needle-Shaped Ascospores as New Members within the Saccharomycetaceae When good bacteria behave badly: a case report of Bacillus Clausii Sepsis in an Immunocompetant Adult Closed Genome Sequence of Bacillus Paralicheniformis Strain Cbmai 1303, a Bacterium Applied for Production of novel polygalacturonase from Bacillus Paralicheniformis Cbs32 and application to depolymerization of Ramie Fiber Biocontrol of Alternaria Alternata and Fusarium Oxysporum by Trichoderma Asperelloides and Bacillus Paralicheniformis in Tomato Plants The Extremophile Anoxybacillus Sp. Pc2 isolated from Brazilian Semiarid Region (Caatinga) Produces a Thermostable Keratinase Integrating multiple genomic technologies to investigate an outbreak of Carbapenemaseproducing enterobacter Hormaechei Seed Biopriming with Pand K-solubilizing Enterobacter Hormaechei Sp. improves the early vegetative growth and the P and K uptake of Okra Improving UV seawater disinfection with immobilized Tio2: study of the viability of photocatalysis (Uv254/Tio2) as seawater disinfection technology First genomic characterization of Blavim-1 and Mcr-9-Coharbouring Enterobacter Hormaechei Isolated from Food of Animal Origin. Pathogens, 9 A comparative study of Candida species diversity among patients with oral squamous cell carcinoma and oral potentially malignant disorders Whole-genome sequencing and mining of protease coding genes in Bacillus Paralicheniformis Mku3, and its degradomics in feather meal medium Transfer of Streptococcus lactis and related streptococci to the genus Lactococcus gen Aromatic and monoterpene alcohol accumulation by Eremothecium Ashbyi strains differing in Riboflavinogenesis Bloodstream infection with Candida Kefyr/Kluyveromyces Marxianus: case report and draft genome Emerging bacterial fish pathogen Lactococcus Garvieae Rtcli 04, isolated from rainbow trout (Oncorhynchus Mykiss): genomic features and comparative genomics. Microbial Pathogenesis Emergence of a multidrug-resistant Enterobacter Hormaechei clinical isolate from Egypt Co-Harboring Mcr-9 and Blavim-4. Microorganisms The prevalence of pathogens causing bovine mastitis and their associated risk factors in 15 large dairy farms in China: an observational study Vitro Antimicrobial Resistance Profiles of Streptococcus Uberis, Lactococcus Spp., and Enterococcus Spp Enterobacter hormaechei subsp. hoffmannii subsp. nov., Enterobacter hormaechei subsp. xiangfangensis comb. nov., Enterobacter roggenkampii sp. nov., and Enterobacter muelleri is a later heterotypic synonym of Enterobacter asburiae based on computational analysis of sequenced Enterobacter genomes Peribronchial connective tissue infection caused by Bifidobacterium Longum and veillonella species mimicking lung cancer Unique hyper-thermal composting process in Kagoshima city forms distinct bacterial community structures Characterization of an extendedspectrum beta-lactamase Enterobacter Hormaechei Nosocomial Outbreak, and Other Enterobacter Hormaechei Misidentified as Cronobacter (Enterobacter) Sakazakii. Microbiology (Reading) Draft genome sequence of enterobacter Hormaechei Strain Mhsd6, a plant endophyte isolated from medicinal plant Pellaea Calomelanos. Microbiol Resoure Announcment Candidemia in children with Malignancies: report from the infection working group of the Hellenic Society of Pediatric Hematology-Oncology Bloodstream infection by Saccharomyces Cerevisiae in two Covid-19 patients after receiving supplementation of Saccharomyces in the Icu Spectrofluorimetric method for the estimation of total lipids in Eremothecium Ashbyii Fungal filaments using nile blue and avoiding interference of Autofluorescent riboflavin Complete genome sequence of Bacillus Paralicheniformis Mdjk30, a plant growth-promoting Rhizobacterium with antifungal activity First report of Enterobacter Hormaechei with respiratory disease in calves Precise species identification for Enterobacter: a genome sequence-based study with reporting of two novel species, Enterobacter quasiroggenkampii sp. nov. and Enterobacter quasimori sp. nov. mSystems Distribution of candidemia in malaysian tertiary care hospital revealed predominance of candida parapsilosis Occurrence of Enterobacter Hormaechei Carrying Blandm-1 and Blakpc-2 in China Taxonomic rearrangement of the Genus Schizochytrium Sensu Lato Based on Morphology, Chemotaxonomic Characteristics, and 18s Rrna Gene Phylogeny (Thraustochytriaceae, Labyrinthulomycetes): emendation for Schizochytrium and Erection of Aurantiochytrium and Oblongichytrium Gen Coproduction of Mcr-9 and Ndm-1 by Colistin-Resistant Enterobacter Hormaechei isolated from bloodstream infection Role of yeasts in the cranberry fruit rot disease complex Bioconversion of lutein by enterobacter hormaechei to form a new compound, 8-methyl-alpha-ionone Does probiotic bacteremia in premature infants impact clinically relevant outcomes? A case report and updated review of literature Peribronchial Connective Tissue Infection Caused by Bifidobacterium longum and Veillonella Species Mimicking Lung Cancer Lactobacillus rhamnosus infection: a single-center 4-year descriptive analysis Lactobacillus acidophilus Endocarditis Complicated by Pauci-Immune Necrotizing Glomerulonephritis A rare bacteria: Lactobacillus Rhamnosus in Pediatric Lung Abscess Lactobacillus endocarditis in a healthy patient with probiotic use. IDCases Prosthetic joint infection after total hip arthroplasty caused by Lactobacillus paracasei Pathogenicity of aerobic bacteria isolated from honeybees (Apis mellifera) in Ordu Province Isolation and Identification of Potentially Pathogenic Microorganisms Associated with Dental Caries in Human Teeth Biofilms. Microorganisms Leuconostoc lactis and Staphylococcus nepalensis Bacteremia Leuconostoc lactis-A Rare Cause of Bacterial Meningitis in an Immunocompromised Host When good bacteria behave badly: a case report of Bacillus clausii sepsis in an immunocompetant adult The prevalence of pathogens causing bovine mastitis and their associated risk factors in 15 large dairy farms in China: an observational study Candida kefyr in Kuwait: prevalence, antifungal drug susceptibility and genotypic heterogeneity Pattern and Frequency of Hospital Acquired Infections in Pediatric Intensive Care Unit at Abha Maternity and Children Hospital, Saudi Arabia Breakthrough fungemia caused by Yarrowia lipolytica in a patient with gastric adenocarcinoma during echinocandin therapy Fatal case of Fungemia by Wickerhamomyces anomalus in a pediatric patient diagnosed in a teaching Hospital from Brazil Molecular identification of Candida isolates by Real-time PCR-high-resolution melting analysis and investigation of the genetic diversity of Candida species Low prevalence of antifungal resistant Candida africana, in the C. albicans complex causing vulvovaginal candidiasis. Heliyon, 6, e03619 High rates of misidentification of uncommon Candida species causing bloodstream infections using conventional phenotypic methods Recurrent vulvovaginal candidosis and cluster analysis of clinical signs and symptoms: a laboratory-based investigation Candidaemia in a tertiary care centre of developing country: Monitoring possible change in spectrum of agents and antifungal susceptibility Risk prediction for candidemia in surgical intensive care unit patients Wickerhamomyces anomalous: a rare cause of fungemia causing febrile neutropenia in acute lymphoblastic leukemia Virulence factors and antifungal susceptibility in candida species isolated from dermatomycosis patients Rare invasive yeast infections in greek neonates and children, a retrospective 12-year study Necessity to identify candida species accurately with minimum inhibitory concentration determination in each case of bloodstream infections Evolution of fungemia in an Italian region A comparative study of Candida species diversity among patients with oral squamous cell carcinoma and oral potentially malignant disorders Bloodstream infection with Candida kefyr/Kluyveromyces marxianus: case report and draft genome Candidemia in children with malignancies: report from the infection working group of the hellenic society of pediatric hematology-oncology Bloodstream infection by Saccharomyces cerevisiae in two COVID-19 patients after receiving supplementation of Saccharomyces in the ICU Distribution of candidemia in Malaysian tertiary care hospital revealed predominance of Candida parapsilosis Role of yeasts in the cranberry fruit rot disease complex Protists/algae Impact of feeding n-3 fatty acids to layer breeders and their offspring on concentration of antibody titers against infectious bronchitis, and Newcastle diseases and plasma fatty acids in the offspring Comparative study on growth, survival and pigmentation of Solea aegyptiaca larvae by using four different microalgal species with emphasize on water quality and nutritional value Comparing docosahexaenoic acid supplementation strategies in terms of broiler tissue enrichment, productivity, and cost Safety of Schizochytrium sp Appendix A -Search strategy followed for the (re)assessment of the suitability of TUs notified to EFSA not present in the current QPS list for their inclusion in the updated list (reply to ToR 3)A.1. The search on PubMed led to 168 hits with the keyword 'hormoachei' 168 hits. All hits were screened for their relevance. Corynebacterium stationisThe search on Scopus led to 17 hits related to Corynebacterium stationis. Among them, seven were considered appropriate for the assessment.A.3. A search for the body of knowledge on L. garvieae was done in the databases mentioned above, considering all years available in these databases, using a range of search terms in relation to food and feed, combined with L. garvieae.lactococcus AND garvieae AND (infect* OR disease* OR pathog* OR mastit*): 277 hits, lactococcus AND garvieae AND cheese: 23 hits lactococcus AND garviae AND (food OR feed OR ferment*): 13 hits lactococcus AND garvieae AND fish: 213 hits lactococcus AND garvieae AND mastit*: 18 hits A.4. Searches were made on Web of Science core collection. '(Eremothecium ashbyi) or (E. ashbyi) or (Eremothecium ashbyii) or (E. ashbyii)' gave 150 hits, which were all scanned on title level. A similar search was also done with the synonym Crebrothecium ashbyi, resulting in no additional hits. A search was made on PubMed with the key word 'Bacillus paralicheniformis'. All 43 hits were screened.A.6. Search in Pub-Med for the terms "caldiproteolyticus" and "caldoproteolyticus" provided eight hits. Five of them were on taxonomy/properties of the organism, while the other three were on description of related, new Anoxybacillus species. Kodamaea ohmeri Search in Pub-Med for the terms "Kodamaea" and "ohmeri" for the years 2019, 2020 and 2021 resulted in 15 papers, 7 papers and 1 paper, respectively. Collectively, out of these 23 papers one is a review, two are previous QPS statements, three are related to bioremediation/biochemistry, seven are related to plant pathology/ecology and 10 are related to (treatment of) opportunistic K. ohmeri infections in mostly immunocompromised human individuals. Other databases, including Web of Science core collection, searched for the same terms gave less papers, but with full overlap with the Pub-Med collection of papers.Appendix B -Protocol for Extensive literature search (ELS), relevance screening and article evaluation for the maintenance and update of list of QPS-recommended biological agents (reply to ToR 2)The protocol for extensive literature search (ELS) used in the context of the EFSA mandate on the list of QPS-recommended biological agents intentionally added to the food or feed (EFSA-Q-2020-00079) is available on the EFSA Knowledge Junction community on Zenodo, at: https://doi.org/ 10.5281/zenodo.5016562Appendix C -Search strategies for the maintenance and update of list of QPS-recommended biological agents (reply to ToR 2)The search strategies for each taxonomic unit (TU), i.e. the string for each TU and the search outcome, are available on the EFSA Knowledge Junction community on Zenodo at: https://doi.org/ 10.5281/zenodo.5016711Appendix D -References selected from the ELS exercise with potential safety concerns for searches July-December 2020 (reply to ToR 2) None. None.Pediococci spp. Propionibacterium spp. None. The list of QPS status recommended biological agents (EFSA BIOHAZ Panel, 2020a) is being maintained in accordance with the mandate of the BIOHAZ Panel (2020-2022), extended for the following years. Possible additions to this list are included approximately every 6 months, with the last Panel Statement (13)