Internal Colonization of Salmonella enterica Serovar Typhimurium in Tomato Plants

Several Salmonella enterica outbreaks have been traced back to contaminated tomatoes. In this study, the internalization of S. enterica Typhimurium via tomato leaves was investigated as affected by surfactants and bacterial rdar morphotype, which was reported to be important for the environmental persistence and attachment of Salmonella to plants. Surfactants, especially Silwet L-77, promoted ingress and survival of S. enterica Typhimurium in tomato leaves. In each of two experiments, 84 tomato plants were inoculated two to four times before fruiting with GFP-labeled S. enterica Typhimurium strain MAE110 (with rdar morphotype) or MAE119 (without rdar). For each inoculation, single leaflets were dipped in 109 CFU/ml Salmonella suspension with Silwet L-77. Inoculated and adjacent leaflets were tested for Salmonella survival for 3 weeks after each inoculation. The surface and pulp of ripe fruits produced on these plants were also examined for Salmonella. Populations of both Salmonella strains in inoculated leaflets decreased during 2 weeks after inoculation but remained unchanged (at about 104 CFU/g) in week 3. Populations of MAE110 were significantly higher (P<0.05) than those of MAE119 from day 3 after inoculation. In the first year, nine fruits collected from one of the 42 MAE119 inoculated plants were positive for S. enterica Typhimurium. In the second year, Salmonella was detected in adjacent non-inoculated leaves of eight tomato plants (five inoculated with strain MAE110). The pulp of 12 fruits from two plants inoculated with MAE110 was Salmonella positive (about 106 CFU/g). Internalization was confirmed by fluorescence and confocal laser microscopy. For the first time, convincing evidence is presented that S. enterica can move inside tomato plants grown in natural field soil and colonize fruits at high levels without inducing any symptoms, except for a slight reduction in plant growth

Pest categorisation of Unaspis citri

The Panel on Plant Health performed a pest categorisation of the citrus snow scale, Unaspis citri (Comstock) (Hemiptera: Diaspididae), for the European Union (EU). This is a well-de fi ned and distinguishable species, native to south-eastern Asia, which has spread to many tropical and subtropical regions. U. citri can be a pest of citrus and has been cited on over 28 different species in 16 plant families. In the EU, U. citri occurs in the Azores. There is uncertainty as to whether it occurs in continental Portugal. Reports of it occurring in Greece and Spain are likely to be invalid and based on interception records from these countries. An old Italian record is a misidenti fi cation. U. citri is listed in Annex IIAI of 2000/29/EC as a harmful organism. The international trade of hosts, as either plants for planting, fruit or cut fl owers, provide potential pathways into the EU. However, current EU legislation prohibits the import of citrus plants for planting from third countries. U. citri is mostly con fi ned to coastal humid tropical areas and does not occur in semi-arid areas that are irrigated. Nevertheless, given that it occurs in the Azores and that there are regional climatic similarities between places where U. citri occurs and climates within the EU, and taking EU host distribution into account, U. citri has the potential to establish in the EU, especially in citrus-growing regions around the Mediterranean where losses in quality and yield of citrus could occur. Phytosanitary measures are available to inhibit the likelihood of introduction of U. citri . Considering the criteria within the remit of EFSA to assess the status as a potential Union quarantine pest (QP), or as a potential regulated non-quarantine pest (RNQP), U. citri meets the criteria assessed by EFSA for consideration as a potential Union QP

Pest categorisation of Oligonychus perditus

The Panelon Plant Health performed a pest categorisation of the spider mite Oligonychus perditus Pritchard and Baker () (Acari, Tetranychidae), for the EU. O.perditus is a well-defined and distinguishable species, native to China, Japan, Korea and Taiwan, and recognised mainly as a pest of Juniperus spp., Chamaecyparis spp. and Platycladus spp. It is absent from the EU and is listed in Annex IIAI of Directive 2000/29/EC. Its host plants, Juniperus spp. and Chamaecyparis spp., are also listed in Annex III of Directive 2000/29/EC. Plants for planting, cut flowers and branches are considered as pathways for this pest, which is also able to disperse naturally with the wind, over rather short distances. O.perditus has repeatedly been intercepted in the EU but does not appear to have established, although a small population of O.perditus survived 8years on a single imported plant in the Netherlands. As the host range of O.perditus coincides with that of the closely related cosmopolitan Oligonychus ununguis, which occurs in the EU, it is quite likely that the presence of O.perditus in the EU would cause little additional damage. Cultural control (sanitation and destruction of infested material) and chemical control (acaricides, e.g. abamectin) are the major control methods. All criteria assessed by EFSA for consideration as a potential quarantine pest are met, though there are some uncertainties regarding impacts. The species is presently absent from the EU, and thus, the criteria for consideration as a potential regulated non-quarantine pest are not met

Pest categorisation of Scirtothrips citri

The Panel on Plant Health performed a pest categorisation of the citrus thrips, Scirtothrips citri (Moulton) (Thysanoptera: Thripidae), for the European Union (EU). This is a well-de fi ned and distinguishable species, occurring in North America and Asia. Its precise distribution in Asia is uncertain. S. citri is a pest of citrus and blueberries and has been cited on over 50 different host species in 33 plant families. Whether all plants reported as hosts are true hosts, allowing population development of S. citri , is uncertain. S. citri feeds exclusively on young actively growing foliage and fruit. It is not known to occur in the EU and is listed in Annex IIAI of 2000/29/EC as a harmful organism. The international trade of hosts, as either plants for planting or cut fl owers, provide potential pathways into the EU. However, current EU legislation prohibits the import of citrus plants for planting. Furthermore, measures aimed at the import of plants for planting in a dormant stage (no young foliage or fruits present) with no soil/growing medium attached, decreases the likelihood of the pest ’ s entry via other hosts. Considering that there are regional climatic similarities where S. citri occurs in the USA with climates in the EU, and taking EU host distribution into account, S. citri has the potential to establish in the EU, especially in citrus and blueberry growing regions around the Mediterranean where quality losses in citrus and yield losses in blueberry could occur. Phytosanitary measures are available to inhibit the likelihood of introduction of S. citri from infested countries. Considering the criteria within the remit of EFSA to assess its status as a potential Union quarantine pest (QP) or as a potential regulated non-quarantine pest (RNQP), S. citri meets with no uncertainties the criteria assessed by EFSA for consideration as a potential Union QP

Hot water treatment of Vitis sp. for Xylella fastidiosa

Following a request from the European Commission, the EFSA Panel on Plant Health (PLH) reviewed Italian technical guidelines and the ANSES (Agence nationale de sécurité sanitaire de l’alimentation, de l’environnement et du travail) opinion on the use of hot water treatment (HWT) on Vitis sp. planting material, assessing its efficacy in the elimination of the xylem-invading bacterial pathogen, Xylella fastidiosa. HWT is a robust and reliable technique used to destroy life stages of pests (insects, nematodes) and to inactivate pathogens (phytoplasma, bacteria, fungi) in dormant plant propagation materials (grapevine and other crops). An effective HWT sanitizes the planting material without affecting plant survival and development. For grapevine, HWT to eliminate the Grapevine flavescence dorée phytoplasma (FD) from planting materials is among the special requirements for the introduction and movement of Vitis sp. to protected zones in the EU. The conditions of 50°C for 45 min, prescribed and recommended to sanitize grapevine planting material against FD, are considered by the Panel to be also effective against X. fastidiosa and its subspecies. Despite uncertainties on variable thermotolerances of the bacteria, a HWT treatment of 50°C for 45 minutes can effectively account for different thermotolerances. It should be noted that the quality of the HWT is subject to the proper application of the operating procedures to guarantee vigorous growth and pathogen freedom of planting material

Pest categorisation of Aschistonyx eppoi

The Panel on Plant Health performed a pest categorisation of the gall midge Aschistonyx eppoi Inouye (1964) (Diptera, Cecidomyiidae), for the EU. A. eppoi is a well‐defined and distinguishable species, native to Japan and Korea, and recognised as a pest of Juniperus chinensis, although our knowledge is solely based on one unique publication. A. eppoi is absent from the EU, and is listed in Annex IIAI of Directive 2000/29/EC. Its host plants, Juniperus spp. are also listed in Annex III of Directive 2000/29/EC. Plants for planting and branches are considered as pathways for this pest. A. eppoi has been intercepted twice (1974; 1975) in the EU and has been eradicated. The pest is likely to affect bonsai plants of J. chinensis if it were to establish in the EU territory. However, as it is unknown whether A. eppoi would attack the Juniperus spp. that occur in the EU, its potential impact on the wild vegetation is also unknown. As the pest originates from areas with warm climates, impact outdoors would affect the southern parts of the EU. Cultural control (destruction of infested material) and chemical control are the major control methods. All criteria assessed by EFSA for consideration as a potential quarantine pest are met, although there are high uncertainties regarding impact. The species is presently absent from the EU, and thus the criteria for consideration as a potential regulated non‐quarantine pest are not met

Pest categorisation of Longidorus diadecturus

The Panel on Plant Health performed a pest categorisation of Longidorus diadecturus (Nematoda: Longidoridae) for the EU. The nematode is a well-defined taxon and was described from Ontario, Canada and later reported from some states in the USA. The nematode is not present in the EU. It is regulated by Council Directive 2000/29/EC, listed in Annex I A I as L. diadecturus Eveleigh and Allen. It is a migratory ectoparasitic nematode species puncturing cells of plant roots thereby able to transmit the nepovirus Peach rosette mosaic virus (PRMV). The pest is found in soil associated with plant species belonging to different families. L. diadecturus is able to cause direct damage to plants, but its main damage is caused by vectoring PRMV. Soil is a potential pathway for this nematode for entry into the EU. The nematode is able to survive adverse conditions, but the virus may not persist inside the nematode for extended periods. Climatic conditions in the EU are similar to those found in the countries where the pest is currently present. Hosts of the nematode (and the associated virus) are, e.g. peaches and grapes; those crops are also widely cultivated in the EU. The nematode only moves short distances (around 1 m) but may be spread with soil moving activities. Measures are available to inhibit entry via soil as such. Entry of the nematode with soil attached to plants for planting that are not regulated is possible. L. diadecturus does satisfy all the criteria that are within the remit of EFSA to assess to be regarded as a potential Union quarantine pest

Pest categorisation of Toxoptera citricida

23The European Commission requested EFSA to conduct a pest categorisation of Toxoptera citricida (Hemiptera: Aphididae), an oligophagous aphid developing and reproducing parthenogenetically on tender leaf and flower flush of citrus (Rutaceae). T.citricida is a taxonomic entity with reliable methods available for detection and identification. It is regulated in the EU by Council Directive 2000/29/EC where it is listed in Annex IIAI as a harmful organism whose introduction and spread into the EU shall be banned. T.citricida is native to tropical regions of Southeast Asia and has spread to most citrus-growing areas worldwide, except California and the Mediterranean basin, causing significant damage to citrus as it is the most efficient vector of the Citrus tristeza virus (CTV). T.citricida occurs in Madeira and, with a restricted distribution, in the north-west of the Iberian Peninsula, mostly on backyard citrus trees. This may have hindered the effectiveness of the official control measures in these areas. There are further phytosanitary measures in place in the EU in order to limit entry via traded commodities. Citrus plants for planting are regulated and are a closed pathway. However, there is uncertainty regarding host status of some non-rutaceous plants on which this aphid has been recorded and so other plant genera may provide additional pathways. The EFSA Plant Health Panelconcludes that the establishment of T.citricida in the main EU citrus growing areas around the Mediterranean would have significant impacts because of its ability to vector CTV. Considering the criteria within the remit of EFSA to assess the status as a potential Union quarantine pest (QP), as a potential protected zone quarantine pest (PZQP) or as a potential regulated non-quarantine pest (RNQP), T.citricida meets with no uncertainties the criteria assessed by EFSA for consideration as a potential Union QP.openopenJeger, Michael; Bragard, Claude; Caffier, David; Candresse, Thierry; Chatzivassiliou, Elisavet; Dehnen‐Schmutz, Katharina; Gilioli, Gianni; Grégoire, Jean‐Claude; Jaques Miret, Josep Anton; Navarro, Maria Navajas; Niere, Björn; Parnell, Stephen; Potting, Roel; Rafoss, Trond; Rossi, Vittorio; Urek, Gregor; Van Bruggen, Ariena; Van der Werf, Wopke; West, Jonathan; Winter, Stephan; Gardi, Ciro; Bergeretti, Filippo; MacLeod, AlanJeger, Michael; Bragard, Claude; Caffier, David; Candresse, Thierry; Chatzivassiliou, Elisavet; Dehnen‐schmutz, Katharina; Gilioli, Gianni; Grégoire, Jean‐claude; Jaques Miret, Josep Anton; Navarro, Maria Navajas; Niere, Björn; Parnell, Stephen; Potting, Roel; Rafoss, Trond; Rossi, Vittorio; Urek, Gregor; Van Bruggen, Ariena; Van der Werf, Wopke; West, Jonathan; Winter, Stephan; Gardi, Ciro; Bergeretti, Filippo; Macleod, Ala

Vitis sp. response to Xylella fastidiosa strain CoDiRO

Following a request from the European Commission, the EFSA Panel on Plant Health assessed a scientific report submitted by the Italian Authorities to the European Commission to support a request to delist Vitissp. from Annex I (‘specified plants’) of the Commission Implementing Decision (EU) 2015/789 of 18 May 2015 to prevent the introduction into and the spread within the Union of Xylella fastidiosa (Wells et al.). The report comprised (i) surveys to detect X. fastidiosa in vineyards located in the epidemic zone of CoDiRO with high numbers of diseased olive trees; (ii) inoculation experiments to infect grapevine with a X. fastidiosa isolate ‘De Donno’ from CoDiRO diseased olives; and (iii) vector transmission experiments with X. fastidiosa infective Philaenus spumarius. The Panel acknowledges the difficulties in providing evidence about this hitherto unknown pathogen/vector/host interaction to support the hypothesis that a plant species cannot be infected with a pathogen. Although field surveys to detect X. fastidiosa in grapevine were negative, there was no supporting information on infective vector populations present in the vineyards. Hence absence of infection pressure cannot be excluded. Furthermore the failure to infect grapevine plants either by artificial inoculation or by vector transmission might be due to inoculation conditions not appropriate to induce infections in grapevine. The detection of X. fastidiosa DNA in inoculated grapevine plants even 12 months after inoculation, although localised at the inoculation points, cannot exclude that the DNA amplified by qPCR was from viable cells. The results presented are coherent and provide converging lines of evidence that grapevine (Vitis vinifera) is not a major susceptible host of X. fastidiosa strain CoDiRO. However, from the experimental evidence it is premature to exclude that systemic infections of V.vinifera and Vitissp. occur and that infections at limited foci could serve as a source of inoculum

Pest categorisation of Scirtothrips aurantii

The Panel on Plant Health performed a pest categorisation of the South African citrus thrips, Scirtothrips aurantii Faure (Thysanoptera: Thripidae), for the European Union (EU). This is a well-de fi ned and distinguishable species, recognised as a pest of citrus and mangoes in South Africa, which has been cited on more than 70 different plants, including woody and herbaceous species. It feeds exclusively on young actively growing foliage and fruit. S. aurantii is not known to occur in the EU and is listed in Annex IIAI of 2000/29/EC as a harmful organism presenting a risk to EU plant health. The international trade of hosts as either plants for planting or cut fl owers provide potential pathways into the EU. However, current EU legislation prohibits the import of citrus plants. Furthermore, measures aimed at the import of plants for planting in a dormant stage (no young foliage or fruits present) with no soil/growing medium attached, decreases the likelihood of the pest entry with such plants. Interceptions have occurred on Eustoma grandi fl orum cut fl owers. Considering climatic similarities between some of the countries where S. aurantii occurs (South Africa, Australia) and the EU, its thermal biology and host distribution in the EU, S. aurantii has the potential to establish, especially in citrus-growing regions of the EU. S. aurantii would most probably breed all year long around the Mediterranean and could cause crop losses in citrus, especially oranges. Phytosanitary measures are available to inhibit the introduction of S. aurantii . Considering the criteria within the remit of EFSA to assess its status as a potential Union quarantine pest (QP) or as a potential regulated non-quarantine pest (RNQP), S. aurantii meets with no uncertainties the criteria assessed by EFSA for consideration as a potential Union Q