European Commission Dg(Sante) 2018-6484 Final Report of an Audit Carried out in Spain from 27 February 2018 to 09 March 2018 In

Ref. Ares(2018)5514253 - 26/10/2018

EUROPEAN COMMISSION DIRECTORATE-GENERAL FOR HEALTH AND FOOD SAFETY

Health and food audits and analysis

DG(SANTE) 2018-6484

FINAL REPORT OF AN AUDIT CARRIED OUT IN SPAIN FROM 27 FEBRUARY 2018 TO 09 MARCH 2018 IN ORDER TO EVALUATE THE SITUATION AND OFFICIAL CONTROLS FOR XYLELLA FASTIDIOSA Executive Summary

This report describes the outcome of an audit carried out by the Directorate-General for Health and Food Safety of the European Commission in Spain from 27 February to 9 March 2018. The objective of the audit was to evaluate the situation of Xylella fastidiosa (Xf), in particular, the implementation of Commission Implementing Decision 2015/789/EU of 18 May 2015 setting out protective measures for its control ("the Decision"), as well as the actions implemented by the Spanish competent authorities in response to the recommendations of audit 2017-6216 performed in June 2017.

Containment efforts within the Balearic Islands are in line with the Decision and provide good levels of containment assurance. On-going survey activities there have revealed four new host species (Calicotome spinosa, Genista lucida, Rhamnus alaternus and Juglans regia) with the infections of each determined as subspecies fastidiosa, and have since been notified to the Commission.

With respect to the most recent outbreak in Alicante, in view of the incomplete 2017 survey and a significant backlog of samples still for testing, the full extent of the outbreak remains unknown. Furthermore, there are major delays in the implementation of eradication measures, primarily due to a number of legal challenges and lack of access to gated plots. Only 4,645 almond trees have been felled out of 52,200. Eradication measures are unlikely to be completed before the start of the next vector flight period. Olives, although listed as a host plant in EU legislation, are not being cut within the 100 m radius of infected plants, and other host species are not appropriately destroyed. Furthermore, due to inherent limitations of the test method for subspecies determination, with a current success rate of only 20%, information on the Xylella subspecies present in the Alicante demarcated area remains inconclusive.

Despite concerted efforts in awareness raising with specific stakeholders and movement controls of host material, given the level of uncertainty with respect to the full extent of the outbreak, as well as delays in implementing eradication measures, the risk for further spread of the disease is high.

The intensity of surveys in Spain outside the demarcated area, especially in crops of main economic importance, has increased considerably, with the exception of the main olive growing region (Andalusia), where it remains disproportionately low.

The report contains recommendations to the National Plant Protection Organisation of Spain to address the shortcomings identified.

I Table of Contents

1 Introduction...... 1 2 Objectives and scope...... 1 3 Legal Basis...... 2 3.1 Relevant EU Legislation/measures...... 2 3.2 International standards...... 2 4 Background ...... 3 4.1 Situation of Xylella fastidiosa in the EU ...... 3 4.2 Situation of Xylella fastidiosa in Spain ...... 3 4.3 Commission audits on Xylella fastidiosa ...... 5 5 Findings and Conclusions ...... 5 5.1 Organisational aspects of the plant health controls ...... 5 5.2 Surveys for the detection of Xylella fastidiosa...... 8 5.3 Balearic Islands – containment zone ...... 10 5.4 Movement control of specified plants ...... 12 5.5 Alicante - eradication zone...... 15 5.6 Movement control of specified plants ...... 21 5.7 Diagnostic Laboratories...... 23 5.8 Follow-up of recommendations from audit report 2017-6216 ...... 26 6 Overall Conclusions ...... 27 7 Closing Meeting...... 27 8 Recommendations ...... 28

II ABBREVIATIONS AND DEFINITIONS USED IN THIS REPORT

Abbreviation Explanation AC Autonomous Community ACCA Autonomous Community Competent Authority BZ Buffer Zone as referred to in Article 4(2) of the Decision CA Competent Authority CCA Central Competent Authority Spanish National Research Council, Institute for Sustainable Agriculture, (Laboratorio CSIS laboratory de Biologia y Ecologia de la microbiota del Suelo del Instituto de Agricultura Sotenible del Consejo Superior de Investigaciones Científicas, CSIC) DA Demarcated Area as referred to in Article 4 of the Decision DG Health and Food European Commission's Directorate-General for Health and Food Safety Safety EPPO European and Mediterranean Plant Protection Organisation EU European Union As referred to by Article 1(b) of the Decision - Plants for planting other than seeds Host plant belonging the genera and species listed in the Commission database of plant susceptible to Xf in the EU territory, IPPC International Plant Protection Convention ISPM International Standards for Phytosanitary Measures IZ Infected Zone as referred to in Article 4(2) of the Decision Plant Health Official Laboratory of the Balearic Islands (Laboratoro Oficial de Sanidad LOSVIB Vegetal de las Islas Baleares) Ministry of Agriculture, Fisheries, Food and Environment (Ministerio de Agricultura y MAPAMA Pesca, Alimentación y Medio Ambiente) MLST Multi Locus Sequence Typing MS Member State of the European Union NPPO National Plant Protection Organisation National Reference Laboratory - for Xf in Spain it is the Laboratory for Bacteriology of NRL the Valencian Institute for Agricultural Research, (IVIA – Instituto Valenciano de Investigaciones Agrarias) PCR Polymerase Chain Reaction (Sistema de Información Geográfica de Parcelas Agrícolas) Electronic agricultural land SIGPAC register As referred to in Article 1(b) of the Decision - Host plants and all plants for planting Specified plant other than seeds belonging to the genera or species listed in Annex I of The Decision ST Sequence Type (Empresa de Transformación Agraria, S.A) - a public holding specialised in the TRAGSA provision of emergency services in agriculture, rural development and nature conservation Xf Xylella fastidiosa

III 1 INTRODUCTION

This audit took place in Spain from 27 February to 9 March 2018. It was part of the published audit programme of the Directorate General for Health and Food Safety of the European Commission (DG Health and Food Safety).

The audit team consisted of two auditors of the DG Health and Food Safety. Representatives of the Ministry of Agriculture, Fisheries, Food and Environment (MAPAMA) accompanied the audit team during the audit in addition to representatives of the Regional Authorities in each of the respective Autonomous Communities visited.

An opening meeting was held on 27 February 2018 at the headquarters of MAPAMA in Madrid, during which the objectives and itinerary for the audit were confirmed and additional information necessary for the conduct of the audit was requested.

Unless specified otherwise, the data in the report was provided by MAPAMA.

2 OBJECTIVES AND SCOPE

The objectives of the audit were to evaluate;

 The situation of Xylella fastidiosa (Xf) and the measures applied by the competent authorities (CAs) for its control, in particular the implementation of Commission Implementing Decision 2015/789/EU (the Decision), and  The actions implemented by the CAs in response to the recommendations of audit 2017-6216.

To meet the objectives the following meetings and visits were carried out:

Meetings, visits No. Comments Competent Central 2 MAPAMA Authorities Regional 2 Balearic Islands and Valencia Plant health control sites Balearic Islands Elimination 1 Infected Zone in Ibiza (Containment Movement 5 One Nursery in Mallorca, two airports (Palma zone) controls de Mallorca and Ibiza), and two sea ports (Palma de Mallorca and Ibiza).

Alicante Surveillance 1 Technical meeting with TRAGSA survey team (Eradication zone) Eradication 4 Infected zone sites (almond plantations) Movement 3 Nurseries/garden centres within the DA controls Stakeholder 1 Representing seven farming, growers and meetings nursery associations.

1 Diagnostic laboratories 4 One visit to the Plant Health Official Laboratory of the Balearic Islands, and further technical meetings with the Official laboratory of bacteriology (Valencia), the National Reference Laboratory for Xf, and the laboratory of the Institute for Sustainable Agriculture (responsible for determining subspecies identification)

The audit focused on measures taken in both Demarcated Areas (DAs), in Alicante and in the Balearic Islands, in particular with respect to eradication (or suppression in the Balearic Islands), surveillance and control on movement of plants out of the DAs.

3 LEGAL BASIS

The audit was carried out under the general provisions of EU legislation, in particular under Articles 21 and 27(a) of Council Directive 2000/29/EC, and in agreement with the National Plant Protection Organisation of Spain (NPPO).

3.1 Relevant EU Legislation/measures

Council Directive 2000/29/EC provides for protective measures against the introduction into, and spread within, the EU of organisms harmful to plants or plant products. Commission Implementing Decision 2015/789/EU of 18 May 2015, as amended ("the Decision"), as regards measures to prevent the introduction into and the spread within the Union of Xf (Wells et al.) provides for detailed rules related to findings or outbreaks of the pathogen. The Decision defines the terms 'host plant' and 'specified plant' in relation to Xf and the applicable measures. Specified plants are listed in Annex I of the Decision and the list of host plants is published in a Commission database:

(https://ec.europa.eu/food/plant/plant_health_biosecurity/legislation/emergency_measures/xyl ella-fastidiosa/susceptible_en). All EU legislation/measures referred to in this report are listed in Annex 1. References to legislation/measures are to the latest amended version, where applicable.

3.2 International standards

Article X(4) of the International Plant Protection Convention (IPPC) establishes that contracting parties should take into account, as appropriate, international standards when undertaking activities related to the Convention. The Decision refers to certain International Standards for Phytosanitary Measures, (ISPMs) issued by the IPPC. The ISPMs of relevance to this audit are listed in Annex 2. Their full text is available on the IPPC website (www.ippc.int).

2 4 BACKGROUND

4.1 Situation of Xylella fastidiosa in the EU

Xf is considered one of the most dangerous plant pathogenic bacteria worldwide. With over 300 different host species identified to date, the bacterium causes a wide range of diseases, with significant economic impact for agriculture, although not all infections result in disease symptoms (asymptomatic infection). Four subspecies of the pathogen (pauca, multiplex, fastidiosa and sandyi) have been identified, each with its own host range. The bacterium is found within xylem tissue and normally spreads by insect vectors of the order Hemiptera (suborder Auchenorrhyncha) that feed on xylem fluid, such as spittlebugs, cicadas and sharpshooters.

Xf was reported for the first time in the EU territory by the Italian authorities in October 2013, in the region of Apulia, affecting mainly olive trees. In July 2015, the French authorities reported the first outbreak of Xf in Corsica (which has since adopted a containment approach to its control) and later in the region of Provence-Alpes Côte d'Azur. In June 2016 the German authorities notified the first occurrence of Xf in Saxony on oleander (Nerium oleander) in a greenhouse. This outbreak has recently been formally confirmed as eradicated.

4.2 Situation of Xylella fastidiosa in Spain

Balearic Islands Spain reported the presence of Xf subspecies multiplex on cherry (Prunus avium) and Polygala myrtifolia plants in a nursery on the island of Mallorca in November 2016. Three different subspecies have since been found infecting a wide range of plant species at different locations across three of the principal islands of this Autonomous Community; Xf subspecies multiplex and fastidiosa in Mallorca, subspecies multiplex in Menorca, and subspecies pauca in Ibiza. No Xf infection has been detected on the island of Formentera. The entire territory of the Balearic Islands was declared a demarcated area (DA) in June 2017. The management of the disease changed to a containment approach with the amendment to the Decision as of December 20171. Figure 1. shows the infected zones as updated in March 2018.

1 Commission Implementing Decision (EU) 2017/2352 of 14 December 2017 amending Implementing Decision (EU) 2015/789 as regards measures to prevent the introduction into and the spread within the Union of Xylella fastidiosa (Wells et al.), O.J. L 336, 16.12.2017, p. 31-44.

3 Figure I. Containment zone for Xf and infected zones in the Balearic Islands (as of March 2018)

Menorca

Ibiza

Mallorca

Formentera

Source: MAPAMA Alicante

Spain notified a second outbreak on 29 June 2017, this time in the province of Alicante (within the autonomous community of Valencia). Further findings have since been reported within the same province with the Demarcated Area (DA) currently established at 87,814 Ha (containing approximately 52,200 almond trees). This outbreak is, so far, exclusively identified as Xf subspecies multiplex (ST6), but, this is based on observation and results of testing, so far restricted to Almond (Prunus dolcis). 153 infected plots have been identified across the province. Figure 2. shows the distribution of infected zones across the current DA as updated in March 2018.

Figure 2. DA for Xf and infected zones in Alicante (as of March 2018)

Source: MAPAMA

4 More information about Xf, including the list of DAs established in the EU, as well as the Commission database of host plants found infected with the bacterium in the EU, is available at the website of DG Health and Food Safety:

(http://ec.europa.eu/food/plant/plant_health_biosecurity/legislation/emergency_measures/ind ex_en.htm).

4.3 Commission audits on Xylella fastidiosa

Reports on previous Xf related audits carried out in Italy, France and Germany, as well as Spain, to evaluate the situation and assess measures applied for its control are available on the website of the DG Health and Food Safety (http://ec.europa.eu/food/audits- analysis/audit_reports/index.cfm).

5 FINDINGS AND CONCLUSIONS

5.1 Organisational aspects of the plant health controls

Legal requirements Articles 1 and 2 of Directive 2000/29/EC (organisational aspects of plant health controls).

Article 3a (contingency plans), Article 6(8) (public awareness), Article 13a (awareness campaigns) and Article 14 (action plan) of the Decision.

Findings

Competent authorities

1. The organisational aspects of plant health controls in Spain are described in sections 1 and 2.11 of the DG Health and Food Safety country profile (http://ec.europa.eu/food/audits-analysis/country_profiles/details.cfm?co_id=ES).

2. The situation regarding the structure and organisation of the relevant authorities and their respective competences and responsibilities has not changed since the previous audit (2017-6216) in June 2017.

3. In summary the central competent authority (CCA), the Single Authority in terms of Council Directive 2000/29/EU, is the Directorate-General of Health and Agricultural Production of MAPAMA. Its Sub-directorate General for Forestry, Plant Health and Hygiene is charged with issues related to outbreaks of harmful organisms at national level, such as coordination, information exchange and international representation. The Sub-directorate General for Health Agreements and Border Controls is responsible for import controls, including inspections related to Xf at the phytosanitary border inspection posts.

5 4. Sixteen of the autonomous communities (AC) of Spain are included in the EU plant health regime. Each AC has a plant health service under the relevant ministry/department of the regional government, which is the CA for plant health at AC level (ACCA). Some ACs also comprise provincial plant health services. ACCAs are responsible for the control of harmful organism outbreaks, including implementation of the measures of the Decision, except for the provisions related to controls at the border inspection posts.

5. In forest areas the AC's forestry and environmental authorities are responsible for the implementation of the measures related to Xf, including visual inspections and sampling as appropriate.

6. In the ACs visited, a significant part of the routine surveillance and eradication work is contracted to TRAGSA, a public holding specialised in the provision of emergency services in agriculture, rural development and nature conservation

National and regional legislation

7. The MAPAMA Decision APM/21/2017 of 20 January 2017 establishes specific measures for the control and movement prevention in relation to Xf, (including the original prohibition of transporting specified plants intended for planting from the Balearic Islands).

Balearic Islands  In addition to the four Regional Resolutions issued by the Regional Minister of Environment, Agriculture and Fisheries of the Balearic Islands outlined in audit report 2017-6212, the Regional Resolution of 10 February 2017 adds a ban on the movement of specified plants from Ibiza (where subspecies pauca is present) to the other Islands of the AC.

Valencia  Six Regional Resolutions, formally establishing the DA and eradication measures to be taken, have been issued and form the legal basis to specifically address the outbreak in Alicante. These were issued on 7 July 2017, 25 July 2017, 31 August 2017, 20 October 2017, 19 December 2017 and 19 February 2018.

8. In addition, Castille-Leon published Instruction 1/FYM/2017 of 2 February 2017 on the establishment of specific prevention measures on Xf.

National Contingency Plan 9. The Spanish national contingency plan for Xf, developed in collaboration with the National Plant Health Committee and ACCAs, has been in place since 2015, and was updated in February 2018 (and is kept under continuous review).

10. The structure of the plan is in line with the relevant provisions of the Decision and includes survey, diagnostic and eradication protocols. However, no specific or general

6 advice or guidance exists with respect to dealing with, or overcoming administrative and/or legal impediments towards swift eradication (with no facility to mobilise expert legal advice or mechanism to address generic petitions brought in front of lower courts).

11. With respect to surveys, the plan takes into account the 2015 Commission guidelines for Xf surveys in the EU territory as a basis for the approach to the national survey, with ISPM 31 used as the basis for visual inspections and sample taking as part of the eradication measures in the 100 m infected zone.

12. The plan also states that ACs can prepare their own contingency plans, which may adapt the national measures to their specific conditions. It requires the elaboration of an action plan in the AC after an outbreak is detected.

13. Both the Balearic Islands and Valencia have produced their own Regional Contingency and Action Plans. The regional contingency plan for the Balearic Islands was updated on 1 February 2018, although it was noted by the audit team that the Valencian regional contingency plan had not been updated since 2016.

14. The CCA also reported that Andalusia, Aragon, Castile-La-Mancha and Catalonia have each produced their own regional contingency plans as a precautionary preparatory step to deal with any potential outbreak.

15. The national contingency plan now incorporates an approach based on ISPM 31 for the sampling of plantations in the national survey outside the DAs. Five main crops are targeted - olive, almond, citrus, vine and oak, with a minimum of 400 samples nationally per crop (however, in reality, many more samples are taken (see Section 5.2)).

Information and publicity / awareness raising related to Xf

16. Information and public awareness raising related activities in each of the DAs are given in the respective sections below.

Diagnostic services for Xf

17. There is at least one official laboratory, designated by each ACCA for Xf analysis. MAPAMA stated that each laboratory uses detection methods according to the EPPO protocol PM 7/24(2), in particular the RT-PCR methods of Harper et al., (2010) (and erratum 2013) and Francis et al. (2006) for routine testing. Further details on the respective laboratories involved in Xf controls in both the Balearic Islands and Valencia are given in Section 5.7.

Research and development related to Xf

18. Spain continues its involvement in various research programmes related to the biology, diagnosis, epidemiology and control of Xf at regional, national and international level (EU, EPPO, etc.). These include, but are not limited to, participation in;

7  EU funded POntE (Pest organisms Threatening Europe; https://www.ponteproject.eu/) and Xfactors (Xylella Fastidiosa Active Containment through a multidisciplinary-Oriented Research Strategy; http://www.xfactorsproject.eu/) research (Horizon 2020) programmes, as well as EUPHRESCO coordinated research (PROMODE (Harmonized protocol for monitoring and detection of Xylella fastidiosa in its host plants and its vectors; https://www.euphresco.net/projects/portfolio);  Further identification of the host range, biology, genetic profile and pathogenicity of Xf found in the DA, with the aim of developing early detection methods and effective containment measures;  Refining existing and developing alternative subspecies identification methods;  Development of appropriate genomic markers for informative DNA fingerprinting of isolates as part of population studies (including tracing outbreak source(s));  Characterisation of microfloral differences between tolerant and susceptible olive cultivars;  Species distribution simulation modelling with respect to potential spread under existing and climate change scenarios;  Detection of infected plants with high resolution remote sensing;  Novel control strategies based on bacteriophage dissemination.

Conclusions on the organisational aspects of the plant health controls 19. Although the legislative tools and the national and regional contingency and action plans provide an appropriate framework for the implementation of the control measures, eradication protocols are not implemented in a timely manner and swift eradication is absent.

20. The Valencian Contingency Plan does not take into account modifications to the National Contingency Plan and latest technical advances.

21. Research activities are in line with policy requirements in support of disease management efforts.

5.2 Surveys for the detection of Xylella fastidiosa

Legal requirements Article 2(3) (detection) Article 3 (annual surveys), Article 6(7) (monitoring in the DA) of the Decision.

8 Findings

Surveys in the territory of Spain outside the DA

22. As mentioned in chapter 5.1 above, the National Contingency Plan adopted a change in approach for plantations based on ISPM 31, focusing on five main crops (olive, almond, citrus, vine and oak), with a minimum sample of 400 samples per crop nationwide.

23. This represented a change from the previous approach, which was one inspection per 10,000 ha for vines, apricots, peaches, nectarines, one inspection per 15,000 ha for olives, citrus, almonds, plums and cherries, and one inspection per 50,000 ha for oak, with at least one sample per inspection.

24. However, based on the figures presented in the 2017 report (2017-6216), it is clear interception and sampling rates are much higher, even with respect to ISPM 31 guidance;

 Nurseries: 2,442 visual inspections and an approximate five fold increase in sampling  Plantations: 11,822 visual inspections and an approximate six fold increase in sampling  Forestry and other: almost tripled inspections and an approximate 40 fold increase in sampling.

Table I. summarises the results of the 2014-2017 surveys.

Table I. Xf surveillance in Spain (2014-2017).

Sites with Number Sites without Number Positives Year Inspections symptoms of samples symptoms of samples (total) Nurseries, garden centres 2014 703 14 4 525 33 0 2015 1,486 52 202 1,404 453 0 2016* 1,514 71 94 1,443 2,031 10 2017* 2,442 109 625 2,313 9,550 0 Plantations 2014 1,236 18 18 1,218 31 0 2015 4,453 71 75 4,382 921 0 2016 5,871 128 241 5,743 1,304* 43 2017* 11,822 1,894 2,689 9,928 6,979 321 Forest sites 2014 204 3 0 201 0 0 2015 516 150 4 366 23 0 2016 686 173 10 513 47 0 2017* 1,905 1,026 1,395 879 1,076 303

9 Sites with Number Sites without Number Positives Year Inspections symptoms of samples symptoms of samples (total) Total 2014 2,143 35 22 2,108 64 0 2015 6,437 273 281 6,164 1,397 0 2016* 8,071 372 345 7,699 3,382 53 2017* 16,149 3,029 4,709 13,120 17,605 624 Insect vectors Number of inspections Number of samples Positives 2014 91 246 0 2015 121 327 0 2016* 21,821 483 0 2017* 2,612 1,762 0 *Includes samples taken in the survey in the DA Source: MAPAMA 25. The Andalusia region represents over 80% of all Spanish olive production. Notwithstanding the overall national increase in inspection and sampling, and based on the 2017 survey statistics provided by the CCA, doubling of inspections in olive plantations in Andalusia remains disproportionately low (approximately 11.5%) compared to all olive inspections in the country.

Conclusions on the surveys for the detection of Xylella fastidiosa 26. The intensity of surveys outside the demarcated area, with respect to total number of inspections and samples taken, increased considerably in 2017, particularly for crops of main economic importance. However, the intensity in the main olive growing region (Andalusia) remains disproportionately low.

5.3 Balearic Islands – containment zone

Legal requirements

Article 7 (control measures), Article 9 (movement ban), Article 11 (movement controls) and Article 13a (awareness campaigns) of the Decision.

Findings

27. With the amendment to the Decision of 14 December 2017, the DA of the Balearic Islands became a containment zone. Although many of the control activities subject to the audit took place before this date, the requirements in Article 7 of the Decision are used for the evaluation of control measures.

Regional legislation and plans

10 28. In addition to the national order of 20 January 2017, inter alia prohibiting movement of specified plants out of the Balearic Islands, the activities on the islands are regulated by regional resolutions. A resolution of 10 February 2017 adds a ban on the movement of specified plants from Ibiza (where the subspecies pauca is present) to the other Islands.

29. There is a regional action plan for the Balearic Islands DA, which was most recently updated on 1 February 2018 to take into account the change to a containment approach. In addition, separate plans have been prepared for the protection of sites with specific cultural, social or scientific value, including four government gene banks of local plant varieties (fig, olive, almond, apricot, pear, apple and grapevine) and the olive trees of the Tramuntana mountain range.

Surveys

30. The 2017 survey was still based on covering the entire DA by inspections in a 1km x 1km grid of which there are 5,600. The strategy is described in the previous audit report. Of the squares, 1,437 were actually inspected and 3,380 samples taken. Of these samples, there were 608 positives. With the seven positives from 2016 and 12 so far in 2018, a total of 627 positives have been found since the discovery of the outbreak. None of the positives were from Formentera, but the authorities do not believe this island to be free of the disease. A total of 18 host species have been found infected, the most numerous are olive, followed by almond and grapevine.

31. For the 2018 season, the approach will be changed to focusing on surveying the above mentioned sites with specific cultural, social or scientific value and their 200m vicinity in grids of 100m x 100m. In addition, selected nurseries will continue to be monitored.

32. Amongst the 18 species found infected, four new host species were identified. These are listed in Table II;

Table II. New host species identified from the 2017 survey on the Balearic Islands.

Species No. individual plants and Subspecies designation ST No. positives Calicotome spinosa 3 plants, 1 positive Subspecies fastidiosa 1 Genista lucida 3 plants, 1 positive Subspecies fastidiosa 1 Rhamnus alaternus 24 plants, 4 positive Subspecies fastidiosa 1 Juglans regia 4 plants, 1 positive Subspecies fastidiosa -* *Not yet determined.

Control measures

33. The control measures are described in detail in the previous audit report (2017-6216). In summary, when a positive is found the owner is notified and given 10 days to carry out the necessary measures or request that the authorities do it free of charge. The latter is usually the case. The infected plants are sprayed with insecticide before being felled and uprooted. The felled material is generally burned on site. In the 100m radius, a survey is conducted with the taking of samples for laboratory analysis.

11 34. The audit team visited a site where an infected olive tree had been found and noted that:

 The tree had been notified in May by the gardener of the residential property. Later a second tree was found. Both had been uprooted and burned;

 A survey had been carried out of all the specified plants in the 100m radius. Eleven samples were taken;

 The inspector stated that the site would be inspected again in six months' time.

35. Of the total of 627 infected plants found as at the time of the audit, 542 had been eliminated. The authorities stated that the outstanding plants were found only recently. In some cases, elimination has been delayed by absentee owners. The fact that only infected trees are cut under containment measures means that there is generally good cooperation from owners.

36. As described in the previous audit report, the authorities have produced a booklet for farmers on preventing the spread of Xf, including recommendations for reducing vector populations by soil tillage, herbicide treatment under trees and by the use of insecticides. As these measures only constitute recommendations, no controls are carried out to ensure their implementation.

Conclusions on Balearic Islands – control measures

37. Survey and controls are largely in line with the EU requirements to a containment zone, which were introduced from 14 December 2017.

5.4 Movement control of specified plants

Legal requirements Article 6(8) (awareness raising and road signs), Article 9 (movement ban), Article 10 (traceability) and Article 11 (official checks) of the Decision.

Findings.

38. There are 184 registered producers, wholesalers or retailers of plants in the Balearic Islands, generally catering for the local market. 170 of these have specified plants. Controls at these establishments are described in the previous audit report. The audit team visited one garden centre and found general consistencies with the findings of the previous audit:

 A poster displayed the restrictions on movement of specified plants out of the islands;  There were regular visits by the plant health authorities;  The business had records of incoming and outgoing material, which were checked during the visits of the plant health authorities;

12  Unlike in the Alicante DA, customers are not requested to sign a declaration about not moving their plants out of the islands. However, if a customer asks about such movement they will be given the information. There have been examples where a sale was not completed because of this.

39. Measures to prevent movement of specified plants out of the Balearic Islands (and from Ibiza to the other islands) are taken at the main points of exit. All the main stakeholders at these locations have been mobilised with training and information material, including the airport authorities, port authorities, security personnel, airline and ferry personnel and the civil guard. The measures centre on information to travellers and transporters in the form of posters, flyers, oral announcements, etc., supplemented by physical checks.

40. The volume of traffic in and out of the islands is substantial. The main air traffic is to Palma de Mallorca with some 28 million passengers arriving (and leaving) in 2017; Ibiza Airport has 8 million. The five national ports receive in the order of 3 million ferry passengers and 1.3 million cruise ship passengers in transit per year. Around 300,000 passenger cars arrive per year. In addition there are arrivals/departures by private recreational boats, which can moor at marinas at the national ports or at the 31 smaller local ports. There are approximately 32,000 such craft movements recorded from these marinas and ports per year.

41. The audit team visited Palma de Mallorca and Ibiza Airports, discussed with relevant authorities and personnel and noted that:

 Posters about the movement restrictions were placed in both arrival and departure halls. Smaller posters and flyers were placed on all the check-in counters. In the departure hall, waste bins were placed in connection with posters for the disposal of plant material. Xf posters were also included in the sequence of electronic notices running for general passenger information;  The airport information desks had flyers and the personnel there were informed about the issue;  A short message in the main languages on the movement restrictions was run in between other messages on the airport PA system;  For passenger luggage, the risk of this containing plants is highest for carry-on luggage. This is scanned 100% as part of the standard security procedures. The security personnel had received training on the restrictions. When plants are found the issue is handed over to the civil guard. The civil guard maintain a catalogue, including pictures, with specified plant species and the possibility to send pictures to the plant health authorities in cases of doubt, but in practice, all plants for planting are intercepted. Discarded plants could be observed in the waste bins at the visited security checks;  For checked-in luggage, detection of plants has also been integrated into the international procedures for security checks. A significant proportion of the luggage undergoes checks where plants can be detected;

13  Since June 2017, there have been 45 interceptions of plants at Palma da Mallorca Airport.

42. The audit team visited the sea ports of Ibiza and Palma da Mallorca, and based on discussions with relevant authorities and personnel noted that:

 At the ferry terminal, posters about restrictions were placed in the ticket/check-in hall and flyers were placed on the counter at the ticket office. In the embarking area, there were also posters and waste bins for disposal of plant material. Some of the ferry company ticket officers were also aware of the restrictions;  The port staff at the entrance of the embarking area that check for drivers' boarding cards stated that they also routinely ask if there are plants in the vehicle, and if so, the driver is given a pamphlet;  The civil guard carry out controls in the passenger vehicle embarking area. They stated that all ferry departures are subject to control and all drivers are asked whether they have plants in their car. In the high season, it is not possible to inspect all vehicles, but some 20%-40% are physically checked with opening of luggage compartments. In low season all vehicles are inspected. The audit team observed this for a ferry going to Barcelona. A rosemary plant was intercepted from a car with tourists from another MS;  The port authorities stated that foot passengers on the ferries are few. Information material and disposal bins are available at the foot passenger embarking area and their luggage is subject to a security check (scanner);  For package carriers, haulage contractors and smaller commercial vehicles (vans) travelling on passenger ferries, the same system and inspection frequencies as for passenger vehicles are applied;  Very little freight leaves the islands. The various authorities stated that much freight arrives, but that trucks and containers generally leave empty. The civil guard stated that when containers/trucks do carry load, they check the bill of lading for the possible presence of plant material and do targeted checks;  The cruise ship operators have been given posters to place in embarking areas. Furthermore, passenger luggage is subject to security check under civil guard supervision;  Recommendation 4 of the previous audit report concerned the implementation of appropriate controls at seaports. For the national ports, this recommendation has now been satisfactorily addressed.

43. The plant health authorities stated that operators of the smaller local ports had been given posters to place in the facilities where yachtsmen check in and out. Leaflets are also handed out as part of these administrative procedures. However, there are no Xf controls carried out on leisure craft traffic.

14 Conclusions on Balearic Islands – movement control

44. For both air and ferry traffic, which constitutes the highest risk for movement of material out of the islands, there are now good controls in place and for this traffic, recommendation 4 from the previous audit has been addressed.

45. The risk from cruise ship and leisure craft traffic is smaller and it has been addressed to a degree. However, for the latter there are no official controls carried out to verify compliance.

5.5 Alicante - eradication zone

Legal requirements Article 4 (establishment of demarcated areas), Article 5 (planting prohibitions), Article 6 (eradication measures) and Article 10 (traceability) of the Decision.

Findings

Establishment of the Demarcated Area and surveying

Establishment of the DA 46. The ACCA established a DA immediately following confirmation of the first positive findings in a 0.5 Ha plantation of mature almond trees (P. dulcis) in the municipality of Guadalest (Alicante province). The national reference laboratory confirmed the positive sample on 29 June 2017 and demarcation was made by a Regional Resolution of 6 July 2017. The Resolution also stipulated the eradication measures to be taken. The outbreak was notified to the Commission on 30 June 2017.

47. The DA was composed of an infested zone (IZ) of a 100 m radius around the infected trees (which in reality was taken as the entire plot from where an infected tree was located) and a 10 km radius buffer zone (BZ), in line with the provisions of the Decision.

48. The audit team noted that the ACCA modifies the borders of DA accordingly if further infections are found outside the IZ, or, in cases where further findings are located outside the DA (of which three have been identified so far), a new IZ of 100 m radius is established.

49. Following the initial outbreak notified on 29 June 2017, the DA has been extended five times to take into account further findings, including groups of findings, of infected plots in various municipalities across the DA. Each finding, or group of findings, is formally recognised by a Regional Resolution declaration, and the DA is modified accordingly. In addition, formal notification of owners is also dependent on publication of a Resolution. Table III collates the dates of findings, notification to the Commission and date of

15 declaration by Regional Resolution, as well as the evolution of the number of infected plots and their locations relative to the initial DA.

Table III. Dates of findings, notification to the Commission and date of declaration by Regional Resolution, as well as the evolution of the number of infected plots and their locations relative to the initial DA. Date of finding Date of Date of Regional No. of infected plots notification to EU Resolution Inside DA Outside DA Total 22 June 2017 29 June 2017 7 July 2017 1 - 1 25 July 2017 25 July 2017 31 July 2017 1 - 1 31 August 2017 31 August 2017 5 September 2017 23 3 26 20 October 2017 20 October 2017 24 October 2017 58 - 58 19 December 2017 20 December 2017 22 December 2017 20 - 20 19 February 2018 19 February 2018 19 February 2018 47 - 47 Totals 150 3 153

50. As at the time of the audit, the DA comprised 87,800 Ha (with amended 5 Km radius BZ). Table IV gives a breakdown of the principal crops (and land use categories) as calculated from Valencian land registry data.

Table IV. Composition of the DA with principal crop species and land use categories. Crop species/category Ha Bushland and Forest 55,000 (Pinus spp.) Fruit species 7,900 Olive trees 4,600 Citrus trees 3,400 Arable land 2,900 Almond trees 1,100

Surveying 51. The original survey as adopted by the ACCA was based on 1Km X 1Km grids within the original 10 Km radius, with 100 m X 100 m grids intended to follow. This approach was not in line with the Decision at that time, which stipulated surveys of 100 m X 100 m grids across the entire BZ.

52. The audit team noted that survey work is carried out by well trained and experienced TRAGSA staff (16 in total) with the aid of an integrated GIS electronic agricultural land register (SIGPAC system) originally developed for farm payments to guide their work.

53. Typically, TRAGSA staff select plots within the 1 Km grids based on host composition (olive, almond, citrus, vine and oak), with emphasis on almonds. At a minimum, one or

16 two plots are selected to survey. If more than one host species is present in the grid, at least one plot of each is also surveyed.

54. TRAGSA stated that it follows the principles as laid down in the Commission guidelines for the survey of Xf in the EU territory, as incorporated into the national and regional contingency plans, with respect to visual inspections and sample taking. TRAGSA further stated that it takes approximately 3-4 days for one staff member to complete the survey within one 1 Km grid.

55. Within the 100 m grids, TRAGSA staff survey all plots of the targeted host species (olive, almond, citrus, vine and oak).

56. However, as of end November 2017, when the survey stopped following almond leaf- fall, only 82% of the 1 Km grids (800 out of 972) and 3.7% of the 100 m grids (1,062 out of 28,892) had been achieved within the 5 Km BZ DA.

57. A total of 7,752 samples from across the Alicante DA were collected from 85 different plant species, the principal amongst these, based on the highest share in sample number, are given in Table V.

Table V. The main plant species sampled across the Alicante DA in 2017. Species No. of Samples Olea europaea 2,302 P. dulcis (almond) 1,710 Citrus spp. 727 Ficus carica 591 Prunus spp. (other than 484 P. dulcis) Vitis vinifera 447 Nerium oleander 186 Rosmarinus officinalis 114 Lavandula spp. 52

58. Vector populations are monitored using an existing extensive network of traps for citrus pests, in addition to a series of further traps positioned in and around, predominantly, olive plantations.

59. As of autumn 2017, the principal vectors found within and around the IZs were Neophilaenus spp. (1,209 (73%)) and Philaenus spumarius (248 (15%)).

Sample analysis 60. As at the time of the audit, of the 7,752 samples collected from the 2017 survey in Alicante, only 4,871 had been tested. Of these, 176 samples, representing 153 plots, tested positive.

17 61. The remaining 2,881 samples still awaited testing, preventing timely identification and eradication of a significant part of the potential outbreaks in the DA.

62. All the 176 positive samples came from almond trees. No other species tested positive for Xf.

63. Of the 176 positive samples, 122, representing the maximum diversity with respect to geographic range within the DA, were forwarded to the laboratory of the Institute for Sustainable Agriculture of the Spanish National Research Council (CSIC laboratory) for subspecies determination.

64. Of the 122 samples, only 25 (20%) have been successfully determined to subspecies level, in all cases Xf sub-species multiplex, and all of the same sequence type (ST6) (see also chapter 5.7 below).

65. With regard to vector analysis, 327 samples were tested. Twenty out of 73 individuals of Philaenus spumarius (27%), and three out of 248 individuals of Neophilaenus spp. (1.2%) tested positive for Xf (clarification on the number of individuals sent for subspecies determination, and subsequent results, was requested from the CSIC laboratory).

Planting prohibitions 66. The ACCA stated that no applications for the planting of host plants in IZs have been received.

Eradication measures 67. In addition to the 153 infected plots identified so far, there is an additional 1,583 associated almond plots located within the surrounding 100 m of the infected plots. The 1,736 plots contain 52,200 almond trees, all of which must be cleared.

68. The ACCA stated that insecticide applications (Lambda cyhalothrin) (pyrethroid class) are used against vectors in anticipation of cutting. However, the audit team noted that insecticide applications, at the locations visited undergoing felling, were performed up to seven weeks previously. Taking into account the properties of the active compound used in this instance, and time lapsed from application, this is not an appropriate phytosanitary treatment prior to the removal of plants as required by Article 6(4).

69. Where accessible, and terrain is relatively level, almond trees are generally uprooted and shredded using an industrial tree mulcher fitted on a hydraulic arm. For more inaccessible areas, such as terraces, or at the request of the owner, a team of chainsaw operators along with a tractor to uproot trees, and a tree chipper, are used. The audit team visited four such infected plots and witnessed tree destruction in action.

70. TRAGSA, outsourced by the ACCA, provides the eradication service (following the guidance as detailed in the Valencian regional contingency plan). There is no charge to the owners. Under optimal conditions, and easy terrain, the industrial tree mulcher can

18 destroy up to approximately 120 trees per day (but typically in the range of 60 to 80 trees), whereas a chainsaw team can achieve considerably less (approximately 30-40 trees per day). If the removal is carried out by the owner, a plant health inspector must be present.

71. Olive trees, although sampled and tested as part of the survey (and so far tested negative), are not cut. Given that Olea europaea is a host species as listed in the Commission database of host plants found to be susceptible to Xf subsp. multiplex in the EU, this is a direct non-compliance with the Directive, which requires removal (Article 6(2)) and destruction (Article 6(5)) of all host plants with a 100m radius of infected plants.

72. Other host species, such as Rosmarinus and Lavandula spp., common in the vicinity of infected trees and in margins along plots, as well as other weed species, are not removed (destroyed or treated with herbicide), but merely trimmed, retaining the ability to regenerate. This is not in line with Article 6(2) of the Decision.

73. Of the total 1,736 plots (representing 52,200 almond trees), which must be cleared, only 78 (representing 4,645 trees) had been cleared at the time of the audit of which only 21 were of the original 153 infected plots.

74. There are significant delays in executing eradication measures in the Alicante outbreak due to a combination of administrative, technical (laboratory testing, see also paragraph 61 above) and legal issues and impediments. In the first instance difficulties can arise in identifying and contacting the owner (e.g. holiday homes, abandoned land), or if plots are owned by more than one person.

75. The ACCA stated that a total of 338 administrative appeals against eradication measures had been made by owners. All had been denied. The administrative process normally requires one month (minimum).

76. With regard to legal issues, these, according to the ACCA, are primarily attributable to four separate legal challenges to halt eradication (three by individual private owners, and one by a municipality) lodged with the local court of first instance in recent months. Each application effectively paralyses the eradication approach with respect to all findings cited in the same formal Regional Resolution being challenged. With respect to these four cases, eradications were stopped from 19 September 2017 until 19 October 2017.

77. Although, according to the ACCA, these four challenges had been effectively rejected, this has resulted in a considerable loss of time with respect to advancing the eradication measures. However, other legal issues, primarily with respect to accessibility, are now in play with 'gated' plots being legally inaccessible under trespass law (Law 39/2015 of Common Administrative Procedure).

78. In such cases, access to these plots requires an application to the court for access authorisation, and further subsequent publication under Regional Resolution and formal

19 publication in the National Official Journal (which in turn adds more time to the process).

79. The ACCA stated that they submitted their first requests for access to such locations on 5 December 2017, which were subsequently rejected on 15 and 24 January 2018. The ACCA further stated that they had re-submitted their requests, with reinforced arguments and clarification of EU law, on 26 February 2018.

80. As at the time of the audit, there were 468 plots (representing 13,335 almond trees) with access restrictions.

81. A breakdown with respect to delays (in days) is given in Table VI.

Table VI. Minimum, maximum and average delays (in days) between sampling and final tree felling.

Reason for delay Minimum Maximum Average Between sampling and lab results 16 157 66 Between lab confirmation and Resolution publication 7 49 29 Between first notification to owner and individual warning 0 146 53 Between individual warning and crop removal 0 117 56 Source: ACCA (Valentia)

Agricultural practices

82. The audit team noted that the Valencian Regional Contingency Plan does not contain specific measures on the application of appropriate agricultural practices for the management of Xf and its vectors for the prevention of spread as required by Article 6(11).

83. However, based on site visits and discussions with the ACCA, the audit team noted that apart from the applications of insecticide against vectors in anticipation of cutting, no other agricultural practices in order to reduce bacterium inoculum sources, and the vector population, are applied.

Trace back of outbreak source

84. The ACCA stated that all nurseries and garden centres within the DA maintain records for at least three years, in particular with respect to particular host species as defined under Art. 10 (2a).

85. The audit team visited three such establishments within the DA and found this to be the case. This is in line with the Decision.

86. However, all findings within the DA of the current outbreak in Alicante are of mature almond trees, approximately 30-40 years old, of the variety Marcona. Given the age of the plantations any specific trace back has been impossible.

20 Conclusions on the eradication measures 87. In view of the incomplete 2017 survey and a significant backlog of samples still for testing, the full extent of the outbreak in Alicante remains unknown.

88. Given the major delays in surveying, laboratory testing and rate of tree felling, eradication measures required by the Decision are unlikely to be completed before the start of the next vector flight period. Given that individual vectors from the 2017 season have been found to be contaminated with Xf, this entails considerable risks for further spread.

89. Olives, although listed as a host plant in EU legislation, are not being cut within the 100 m radius of infected plots and may negate eradication measures.

90. Other listed host species within defined IZs are not fully destroyed. Their regeneration may contribute to inoculum persistence in the IZs. Furthermore, the long delay between phytosanitary treatment against vectors and removal of host plants presents a risk that inoculum is dispersed by contaminated vectors during felling.

91. The absence of the application of appropriate agricultural practices may contribute to the further spread of the bacterium within the DA.

92. The source of the outbreak remains unknown.

5.6 Movement control of specified plants

Legal requirements Article 6(8) (awareness raising and road signs), Article 9 (movement ban), Article 10 (traceability) and Article 11 (official checks) of the Decision.

Findings.

Awareness raising

93. There is a high level of engagement with stakeholders both within and outside the Alicante DA. A total of 31 public presentations have been given by the ACCA to municipalities, nine within the DA (some under challenging conditions, resulting in the abandonment of two), and 24 outside the DA since the outbreak occurred in June 2017.

94. Further meetings and presentations were made with technical services of the regional government, local government official (mayors), co-operatives and associations, and to other ACCAs.

95. The ACCA maintains a website (https://www.xylella.es/), developed in conjunction with the Colegio Oficial de Ingenieros Agronomos de Levante which contains detailed information about the pathogen, approach to control, regional legislation, compensation

21 rules and the Regional Contingency Plan (linked to the National Contingency Plan). It also provides a contact form in cases of suspected finding by growers, or for other questions related to Xf from the public, in addition to the central ACCA emergency telephone number.

96. Furthermore, in addition to social media and specific plant health bulletins to stakeholders, in the form of e-mails, the ACCA has prepared posters and printed 10,000 leaflets (addressed to the general public) for wide-scale dissemination. In particular, these are displayed and distributed at garden centres (in addition to the customer declaration form, see paragraph 99 below) or ports and airports (even if outside the DA).

Road signs

97. Road signs are strategically placed at entries to the DA. However, they are small and the message on the signs is not very clear.

Controls in nurseries

98. The ACCA informed the audit team that there are no registered nurseries or garden centres in any of the IZs. Of the 28 nurseries in the DA, all are located within the BZ (five of which are currently not commercially active)

99. The audit team visited three nurseries, which were both producers and traders, with outlets in the BZs and noted that:

 A movement ban out of the DA was introduced quickly, although with some delay in implementation with nurseries and garden centres, where action was taken only when visited by inspectors (approximately a one month delay);  Each establishment sourced their plants for planting from mainland Spain;  The ACCA inspects each nursery at least once a year in order to authorise them for issuing plant passports. Since the outbreak in June 2017, additional controls related to Xf have been performed;  Each nursery/garden centre visited had a copy of the Decision and associated lists of host and specified plants. Staff were found to be knowledgeable and fully aware of the measures implemented after the outbreak of Xf and their obligations regarding movement control and kept detailed records on production and incoming stock as part of their plant pass porting obligations;  In each nursery, high quality posters and flyers, specifically highlighting movement restrictions for specified plants, were clearly displayed;  The Valencian ACCA, in cooperation with retailers, has developed and implemented a system of customer declaration forms, to be completed and signed by individual private clients on the purchase of specified species. This serves to both raise awareness of the movement restrictions to limit private distribution outside the DA, and to provide a record in case of the need to trace forward.

22 100. At the time of the audit, the Valencian ACCA had received no requests from nurseries for the application of the derogation provided by Article 9(2) of the Decision.

101. Furthermore, there were no applications for derogations for planting host plants in the IZs (see also paragraph 66).

Movement control of specified plants at airports, ports

102. As at the time of audit, the DA did not extend to any ports or airports.

Road controls

103. The CCA informed the audit team that since the start of the outbreak the civil guard has conducted 35 road controls on vehicles leaving the DA.

Conclusions on the movement control of specified plants 104. There is an extensive awareness raising programme with particular emphasis on stakeholders. The customer declaration initiative implemented in nurseries within the BZ further raises the awareness of movement restrictions to limit private distribution outside the DA.

105. Plant passport, and specific Xf controls in nurseries provide a level of assurance that specified plants are not leaving the DA.

106. Road signs are both small and lack prominence, as well as contain a not so clear message, whilst the number of road controls, considering the duration of the outbreak, is low.

5.7 Diagnostic Laboratories

107. The LOSVIB continues to act as the regional laboratory for the testing of samples for Xf in the Balearic Islands in support of containment measures, whereas the Official laboratory of bacteriology (Agricultural Production Safety and Control Service (Valencia)) is the designated laboratory for the testing of samples for Xf from the eradication zone from Alicante. Both laboratories continue to employ two distinct RT- PCR methods (Harper et al., 2010 (and erratum 2013) and Francis et al. 2006) in parallel for routine testing in line with the EPPO diagnostic protocol PM 7/24(2) and the Commission Database of validated tests for the identification of Xylella fastidiosa and its subspecies as referred to in Article 3(2) of the Decision. The LOSVIB confirmed a maximum throughput of 200 samples per week, whilst the Official laboratory of bacteriology (Valencia) reported a maximum capacity of 500 samples per week.

108. Although the previous audit (2017-6216) considered a capacity of 300 samples per month as insufficient by the LOSVIB in support of an eradication programme, the change to a containment regime, and the overall increase in monthly capacity, as noted

23 by the current audit team, places this laboratory in a better position to meet its diagnostic support obligations.

109. However, despite the claimed capacity of the Official laboratory of bacteriology (Valencia) to process a maximum 500 samples per week, it is unlikely this facility, irrespective of proposed staff increases, can realistically achieve such throughput, and given the current backlog of 2,881 samples, it is doubtful that it will be able to adequately support the timely implementation of the provisions of Article 6(3) and 6(7) of the Decision.

110. On inspection of the LOSVIB laboratory in the Balearic Islands, the audit team noted that chemical components for the RT-PCR tests are prepared in the same part of the laboratory where samples are received and initially processed. This is not recommended practice within a PCR laboratory as it may constitute a source of cross contamination and possible false positives.

Confirmatory checks (Recommendation 5 of audit 2017-6216) 111. As part of addressing Recommendation 5 of audit 2017-6216 (Consider extending the quality control of the laboratories testing Xf in the form of confirmatory test on each positive and on a proportion of negatives plant samples, in order to provide additional guarantees as to the reliability of the tests.), both laboratories send the following to the NRL for confirmatory testing:

 Approximately 1% of their negative samples (both plant material for re-extraction by the NRL and DNA extracts). The NRL performs the same set of RT-PCR tests on 10-1 and 10-2 serial dilutions of the DNA extracts. As at the time of the audit, there had been no reported divergence in confirmatory testing of negative samples between the NRL and regional laboratory who supplied the material;  With respect to confirmatory testing of positive samples, given the change to containment in the Balearic Islands, only positive samples from novel findings are sent to the NRL. However, for the eradication effort in Alicante, the Official laboratory of bacteriology (Valencia) forwards all positive samples for confirmatory testing the NRL. The NRL stated that as at time of the audit, confirmatory results were 99% consistent. In cases of discrepancy, the NRL requests new samples from the same tree (or host).

112. Although this is considered generally satisfactory, the audit team considers the 1% negative samples sent for confirmatory, as compared to 100% confirmatory testing for positive samples, as disproportionately low.

Proficiency tests (Recommendation 6 of audit 2017-6216)

113. As part of addressing Recommendation 6 of audit 2017-6216 (Consider organising proficiency tests at regular intervals with participation of each laboratory which tests official samples taken in Spain for the presence of Xf, in order to provide additional guarantees as to the reliability of the tests.), the NRL organised a proficiency test of both

24 RT-PCR methods (Harper et al., 2010 (and erratum 2013) and Francis et al. 2006) in November 2017 as follows:

 21 participating laboratories;  36 blind samples of different plant hosts (almond, olive, citrus and Polygala) spiked with different concentrations of inactivated bacteria (including positive and negative controls);  Each participating laboratory used its own DNA extraction procedure;  Assessment on specificity, sensitivity and reproducibility.

114. Of the 21 laboratories, 76% (16) achieved 100% specificity (the same for reproducibility) and 20 achieved 100% sensitivity (one achieving 95%).

115. Additional blind samples were resent to the five laboratories that did not achieve anticipated results to re-test. Upon re-testing, all achieved 100% sensitivity, and three achieved 100% with respect to specificity. Of the two laboratories that did not achieve 100% specificity, one achieved 83%, the other 60%.

116. The NRL reported to the audit team that both the LOSVIB and the Official laboratory of bacteriology (Valencia) laboratories scored 100% on all assessment criteria in the proficiency test.

117. Given the activity of the NRL, and associated laboratories providing official testing, through the organisation and results of the November 2017 proficiency test, the audit team considered the proficiency of the laboratories applying the preferred tests for the presence of Xf as satisfactory.

Subspecies determination

118. The laboratory of the Institute for Sustainable Agriculture of the Spanish National Research Council (CSIC laboratory) is responsible for subspecies identification. The laboratory applies the multi locus sequence typing (MLST) method of Yuan et al., (2010), as recommended by EPPO diagnostic protocol PM 7/24(2), based on seven specifically selected “housekeeping genes”.

119. Based on discussions with the responsible scientist from the CSIC laboratory, the audit team noted that:

 Success of the MLST method is limited; only 20% of NRL confirmed positive samples from the Alicante outbreak (25 out of 122), and 29% of Balearic Island material (78/267) were successfully determined at sub-species level (including ST definition) (audit 2017-6216 had reported a slightly better success rate of 40% for Balearic Island samples during the previous year);  An alternative two gene approach, employing two distinct genes not part of the Yuan et al. (2010) MLST method, yielded slightly improved subspecies results: 28% for

25 Alicante material, and 31% for Balearic Island material. However, this approach does not permit ST determination;  The CSIC laboratory continues to research improvements to the subspecies determination method, including through international cooperation.

Conclusions on diagnostic laboratories 120. The large number of untested samples from the 2017 survey in the Alicante DA reflect issues with respect to throughput capacity and sample management in the Official laboratory of bacteriology (Valencia), and in turn uncertainty regarding the accuracy and full extent of the DA as currently defined.

121. Confirmatory tests by the NRL of all positive samples from the Alicante outbreak, and representative novel findings from the Balearic Islands, provides a good level of technical assurance of laboratory results. Although confirmatory testing of negative results is disproportionately low, Recommendation 5 of the previous audit has been satisfactorily addressed.

122. The majority of regional testing laboratories have proved proficient for the successful application of RT-PCR testing methods for Xf. As such, Recommendation 6 of the previous audit has been satisfactorily addressed.

123. The low level of successful subspecies determination means, in particular with respect to the Alicante outbreak, the range of possible subspecies across the IZs remains unknown.

5.8 Follow-up of recommendations from audit report 2017-6216

With regard to follow-up of the six recommendations made in audit report 2017-6216, which at the time focused exclusively on eradication of the outbreak on the Balearic Islands, it is noted that;

 Recommendation 1, with the exception of olive trees in Andalusia, this recommendation has been largely satisfactorily addressed (see conclusion 26) and is further covered by Recommendation 2 in the current report;  Recommendations 2 and 3, since publication of the amendment to the Decision of 14 December 2017 the DA of the Balearic Islands became a containment zone (see paragraph 27), and as such both recommendations have become redundant;  Recommendation 4, has been satisfactorily addressed as referred to in paragraph 42 and conclusion 44;  Recommendation 5, has been satisfactorily addressed as referred to in paragraphs 111 and 112 and conclusion 122;

26  Recommendation 6, has been satisfactorily addressed as referred to in paragraph 113 and conclusion 122.

6 OVERALL CONCLUSIONS

7 CLOSING MEETING

A closing meeting was held on 9 March 2018 at the headquarters of MAPAMA in Madrid, with participation of representatives of the CCA, as well as the ACCAs via video conference. This included the forestry authorities and representation from the CSIC laboratory, during which the main findings and preliminary conclusions of the audit team were presented.

During the closing meeting the CCA generally accepted the findings and preliminary conclusions of the audit team concerning the non-compliances, highlighting actions to immediately address some of them.

27 8 RECOMMENDATIONS

The NPPO of Spain is recommended to:

No. Recommendation 1. Ensure that the Valencian Regional Contingency Plan is reviewed in line with the guidance and position of the National Contingency Plan, incorporating the latest information regarding the biology and control of Xylella fastidiosa. Article 3a(3) of Decision 2015/789/EU requires that MSs evaluate and review their contingency plans as necessary.

The recommendation is based on conclusion No. 20

Associated finding No.13

2. In line with Article 3(1) of Decision 2015/789/EU, ensure that the intensity of surveys for Xylella fastidiosa outside the demarcated area, in particular in the principal olive growing regions, is proportionate to the size of production and disease threat.

The recommendation is based on conclusion No.26

Associated finding No.25

3. In line with Article 11 of Decision 2015/789/EU, review the risk of movement of host material via leisure craft traffic from the Balearic Islands with a view to the implementation of official controls to verify compliance.

The recommendation is based on conclusion No. 45

Associated findings Nos. 40 and 43

4. Put in place the measures to ensure the immediate removal of the plants specified in Article 6(2) of Decision 2015/789/EC, including complete felling of all infected plants and all other host and symptomatic plants within a radius of 100m around infected plants within the IZs as a matter of urgency, including those IZs where only almonds have been cleared.

The recommendation is based on conclusion Nos. 19, 88, 89 and 90

Associated findings Nos. 10, 67, 71, 72, 73 and 74

5. In line with Article 6(4) of Decision 2015/789/EU, apply appropriate phytosanitary treatments prior to the removal of host plants against vectors, and plants that may act as hosts of vectors, in order to limit the spread of the disease.

28 The recommendation is based on conclusions No.90

Associated finding No. 68

6. In line with Article 6(11) of Decision 2015/789/EU, apply appropriate agricultural practices as an integral part of the management of the bacterium and its vectors, in particular with respect to limiting the spread of the disease.

The recommendation is based on conclusions No.91

Associated findings Nos. 82 and 83

7. In line with Article 6(8) of Decision 2015/789/EU, increase the size and prominence of road signs highlighting the Alicante DA, and implement a more extensive programme of road controls to help prevent movement of specified plant material out of the DA (in line with Article 11).

The recommendation is based on conclusion No. 106

Associated findings Nos. 97 and 103

8. For an appropriate implementation of Articles 3 and 6 of Decision 2015/789/EU, consider mobilising laboratory capacity to test all outstanding samples from the 2017 Alicante DA survey with immediate effect. Furthermore, take necessary measures to ensure availability of sufficient laboratory capacity, capable of processing all samples anticipated to arise from the 2018 survey.

The recommendation is based on conclusion Nos. 87 and 120

Associated findings Nos. 60, 61 and 109

9. For an appropriate implementation of Articles 3 and 4 of Decision 2015/789/EU, explore all necessary technical approaches towards overcoming the low success rate of the MLST subspecies determination method, including, where possible, the incorporation of technical modifications to the existing protocol, towards improved success rates. The recommendation is based on conclusion No. 123 Associated finding No. 119

The competent authority's response to the recommendations can be found at:

http://ec.europa.eu/food/audits-analysis/rep_details_en.cfm?rep_inspection_ref=2018-6484

29 ANNEX 1 – LEGAL REFERENCES

Legal Reference Official Journal Title Dir. 2000/29/EC OJ L 169, 10.7.2000, Council Directive 2000/29/EC of 8 May p. 1-112 2000 on protective measures against the introduction into the Community of organisms harmful to plants or plant products and against their spread within the Community

Dec. 2015/789/EU OJ L 125, 21.5.2015, Commission Implementing Decision (EU) p. 36–53 2015/789 of 18 May 2015 as regards measures to prevent the introduction into and the spread within the Union of Xylella fastidiosa (Wells et al.) ANNEX 2. RELEVANT INTERNATIONAL STANDARDS

International Title Standard

ISPM No. 9 International Standard on Phytosanitary Measures Publication No 9, Guidelines for pest eradication programmes, Food and Agriculture Organisation, Rome, Rome, Adopted 1998; published 2016, https://www.ippc.int/en/publications/611/

ISPM No. 14 International Standard on Phytosanitary Measures Publication No 14, The use of integrated measures in a systems approach for pest risk management, Food and Agriculture Organisation, Rome, Rome, Adopted 2002; published 2016, https://www.ippc.int/en/publications/607/

ISPM No. 31 International Standard on Phytosanitary Measures Publication No 31, Methodologies for sampling of consignments, Food and Agriculture Organisation, Rome, Adopted 2008; published 2016 https://www.ippc.int/en/publications/588/