Commission Implementing Regulation (EU) 2022/1193 of 11 July 2022 establishing measures to eradicate and prevent the spread of Ralstonia solanacearum (Smith 1896) Yabuuchi et al. 1996 emend. Safni et al. 2014

Article 1

Subject matter

This Regulation establishes measures for the purpose of eradicating Ralstonia solanacearum (Smith 1896) Yabuuchi et al. 1996, emend. Safni et al. 2014, the cause of the potato brown rot, and prevent its spread within the Union territory.

Article 2

Definitions

For the purposes of this Regulation, the following definitions apply:

(1) ‘specified pest’ means Ralstonia solanacearum (Smith 1896) Yabuuchi et al. 1996 emend. Safni et al. 2014;

(2) ‘specified plants’ means plants of Solanum tuberosum L. (potato), other than seeds, and plants, other than fruits and seeds, of Solanum lycopersicum (L.) Karsten ex Farw (tomato);

(3) ‘solanaceous host plants’ means wild and cultivated plants of Solanaceae;

(4) ‘volunteer specified plants’ means specified plants which appear in the places of production without having been planted;

(5) ‘tubers intended to be planted in their place of production’ means tubers produced in a specific place of production which are intended to permanently remain in that place and are not intended to be certified.

Article 3

Annual surveys

The competent authorities shall carry out annual surveys for the presence of the specified pest on the specified plants in their territory, in surface water used for irrigation of the specified plants, and in liquid waste, in accordance with the following requirements:

(a) as regards tubers other than those for planting, the surveys shall comprise: (i) sampling from tuber lots in store or from the growing crop, as late as possible between desiccation of haulms and harvest; (ii) visual inspection of the growing crop, where it is possible to visually identify symptoms of the specified pest, and visual inspection of cut tubers in the cases where that inspection is suitable to detect symptoms of the specified pest;

(b) as regards tubers for planting, other than those intended to be planted in their place of production, surveys shall systematically comprise visual inspection of the growing crops and of lots in store, sampling in store or sampling from the growing crops as late as possible between desiccation of haulms and harvest;

(c) as regards tubers intended to be planted in their place of production, the surveys shall be performed on the basis of the identified risk concerning the presence of the specified pest and shall comprise: (i) sampling from tuber lots in store or from the growing crop, as late as possible between desiccation of haulms and harvest; (ii) visual inspection of the growing crop where it is possible to visually identify symptoms of the specified pest, and visual inspection of cut tubers in the cases where that inspection is suitable to detect symptoms of the specified pest;

(d) as regards plants of tomato, surveys shall comprise visual inspection, at appropriate times, of at least the growing crop at the place of production of plants intended for replanting;

(e) as regards solanaceaous host plants, other than the specified plants, and surface water and liquid waste, surveys shall be conducted in accordance with appropriate methods and, where appropriate, samples shall be taken.

Article 4

Measures in case of suspicion of the presence of the specified pest

Pending the results of the detection tests, the competent authority shall:

(a) prohibit the movement of the specified plants from all crops, lots or consignments from which the samples have been taken, except the specified plants under its control for which it has been established that there is no identifiable risk of the specified pest spreading;

(b) trace the origin of the suspected presence;

(c) carry out official control of the movement of any specified plants, other than those referred to in point (a), produced on the place of production from which the samples referred to in point (a) were taken;

(d) prohibit the use of surface water on specified plants and on other cultivated solanaceous host plants until the confirmation or refutation of the presence of the specified pest in the surface water, except where it allows the use of surface water on tomatoes and other cultivated solanaceous host plants grown in greenhouses, on condition that water is disinfected through appropriate methods authorised by the competent authority.

Pending the results of the detection tests, the competent authority shall ensure that all of the following elements are retained and appropriately conserved:

(a) all remaining tubers sampled and, wherever possible, all remaining plants sampled;

(b) remaining specified plant extracts, DNA extracts and additional prepared material for the test;

(c) the pure culture, when appropriate;

(d) all relevant documentation.

Article 5

Measures in case of confirmation of the presence of the specified pest

Where the presence of the specified pest is confirmed on specified plants, the competent authority shall, without delay, take all the following measures:

(a) establish an investigation to determine the extent and primary source(s) of infection in accordance with Annex III, with further tests in accordance with Article 4(1), on at least all clonally related stocks of tubers for planting;

(b) establish a demarcated area, which shall consist at least of an infested zone containing all of the following elements: (i) the specified plants, consignments and/or lots, vehicles, vessels, stores, or units thereof from which an infected specified plant sample was taken, any other objects including packaging material, and the machinery used in production, transport or storage of those specified plants, and, where appropriate, the place(s) of production or the production site(s) where those specified plants were grown or harvested; (ii) all types of items listed in point (i) determined to be probably infected by the specified pest, through pre- or post-harvest contact, or through simultaneous production, irrigation or spraying steps with the infected specified plants and taking into account the elements listed in point 1 of Annex IV;

(c) establish, where necessary to address the phytosanitary risk, a buffer zone around the infested zone taking into account the elements of a possible spread of the specified pest as referred to in point 2 of Annex IV;

(d) designate: (i) the items listed in point (b)(i) as infected; (ii) the items listed in point (b)(ii) as probably infected.

Where the presence of the specified pest is confirmed on crops of solanaceaous host plants other than the specified plants, and where production of the specified plants is identified at risk, the competent authority shall take the following measures:

(a) establish an investigation to determine the extent and primary source(s) of infection in accordance with Annex III, with further tests in accordance with Article 4(1), on at least all clonally related stocks of tubers for planting; and

(b) establish a demarcated area, which shall consist of an infested zone.

The infested zone shall contain the following:

(a) the host plants from which the infected sample was taken;

(b) the host plants liable to be infected by the specified pest and designated as probably infected, through pre- or post-harvest contact, or through simultaneous production, irrigation or spraying steps with the infected host plants.

The competent authority shall designate:

(a) the host plants referred to in the second subparagraph, point (a), as infected;

(b) the hosts plants referred in the second subparagraph, point (b), as probably infected.

Where the presence of the specified pest is confirmed in surface water, in liquid waste discharges from industrial processing or from packaging premises handling specified plants, or on associated wild solanaceous host plants, and where production of the specified plants is identified at risk through irrigation, spraying or flooding with surface water, the competent authority shall take the following measures:

(a) establish an investigation in accordance with Annex III, including a survey at appropriate times on samples of surface water and liquid waste, and on wild solanaceous host plants if present, in order to determine the extent of the infection; and

(b) establish a demarcated area containing an infested zone taking into account the elements of a possible spread of the specified pest as referred to in point 2 of Annex IV.

The infested zone shall contain the following:

(a) the surface water from which the infected sample(s) was (were) taken;

(b) the surface water liable to be infected, taking into account the elements listed in point 2(b)(ii) and (iii) of Annex IV.

The competent authority shall designate:

(a) the surface water referred to in the second subparagraph, point (a), as infected;

(b) the surface water referred to in the second subparagraph, point (b), as probably infected.

The competent authorities shall ensure that all of the following elements are retained and properly conserved:

(a) the material specified in Article 4(3) until at least the completion of all tests;

(b) the material related to the second detection test and to the identification tests when appropriate, until the completion of all tests;

(c) if applicable, the pure culture of the specified pest, until at least one month after the notification procedure under paragraph 5.

Article 6

Measures for eradicating the specified pest

Where specified plants have been planted before they are designated as infected, the planted material shall be immediately destroyed or disposed of in another way in accordance with point 1 of Annex V. The production site(s) where the infected specified plants have been planted shall be designated as infected. A demarcated area shall be established in accordance with Article 5(2), point (b).

Where specified plants have been planted before they are designated as probably infected, the planted material shall be immediately destroyed or the measures as specified in point 2 of Annex VI, shall apply. The production site(s) where the probably infected specified plants have been planted shall be designated as probably infected. A demarcated area shall be established in accordance with Article 5(2), point (b).

Article 7

Specific testing measures for tubers for planting

Article 8

Entry into force

This Regulation shall enter into force on the third day following that of its publication in the Official Journal of the European Union.

This Regulation shall be binding in its entirety and directly applicable in all Member States.

ANNEX I

Scheme for the tests to be performed pursuant to Articles 3, 4, 5, and 7

1. GENERAL PRINCIPLES ON THE PRESENCE OF THE SPECIFIED PEST

1.1.The presence of the specified pest is suspected where a positive result is obtained in the first detection test performed on the specified plant, or on water samples.

1.2.The presence of the specified pest is confirmed in the following cases:

(a) where the first or the second detection test is a selective isolation resulting in colonies with typical morphology, and positive results are obtained in two identification tests performed on the colonies;

(b) where the first and second test are tests other than a selective isolation, and positive results are obtained in two identification tests after the sample has been subject to selective isolation resulting in colonies with typical morphology.

One of those two identification tests shall be a test referred to in points 2.2(e), (f) and (g).

2. TESTS

The detection tests shall be such as to consistently detect at least 10⁴ cells/ml resuspended pellet acquired from asymptomatic samples.

The second detection test shall be based on different biological principles or different nucleotide regions than the first detection test.

The detection tests are the following:

(a) immunofluorescence tests as described in international diagnostic standards;

(b) isolation of the specified pest on the semi-selective growth medium mSMSA, as described in international diagnostic standards;

(c) conventional PCR test using the primers of Pastrik et al., (2002) (¹), as described in international diagnostic standards;

(d) TaqMan® Real-time PCR tests using primers and probes of: (i) Weller et al. (2000) (²), as described in international diagnostic standards; (ii) Vreeburg et al. (2016) (³) (using a TaqMan® probe modified from the original probe described by Weller et al. (2000)), as described in international diagnostic standards; (iii) Vreeburg et al. (2018) (⁴) (so-called NYtor test), as described in international diagnostic standards; (iv) Massart et al. (2014) (⁵), as described in the international diagnostic standards;

(e) LAMP (loop-mediated isothermal amplification) test using the primers of Lenarčič et al. (2014) (⁶) (only for symptomatic plant material), as described in international diagnostic standards.

The identification tests are the following:

(a) immunofluorescence tests as described in international diagnostic standards;

(b) conventional PCR tests using the primers of Pastrik et al. (2002), as described in full detail in international diagnostic standards;

(c) TaqMan® Real-time PCR tests using primers and probes of: (i) Weller et al. (2000), as described in international diagnostic standards; (ii) Vreeburg et al. (2016) (using a TaqMan® probe modified from the original probe described by Weller et al. (2000)) as described in full detail in international diagnostic standards; (iii) Vreeburg et al. (2018) (so-called NYtor test), as described in international diagnostic standards; (iv) Massart et al. (2014), as described in the international diagnostic standards;

(d) LAMP (loop-mediated isothermal amplification) test using the primers of Lenarčič et al. (2014), as described in international diagnostic standards;

(e) phylotype-specific multiplex conventional PCR test (Opina et al. (1997) (⁷); Fegan & Prior (2005) (⁸)), as described in international diagnostic standards;

(f) DNA barcoding (Wicker et al. (2007) (⁹), as described in international diagnostic standards;

(g) MALDI-TOF MS (van de Bilt et al. (2018) (¹⁰)), as described in international diagnostic standards.

3. FLOW CHARTS OF PROCEDURES

^a Isolation can be used as the first or the second detection test. If the presence of the specified pest is suspected on the growth medium, colonies shall be purified to obtain pure cultures on which two identification tests shall be performed.

^b One of these two identification tests shall be a test referred to in points 2.2(e), (f) and (g). Positive results in the two identification tests are required to confirm the presence of the pest.

^c The third detection test shall be based on different biological principles or different nucleotide regions.

^a One of these two identification tests shall be a test referred to in points 2.2(e), (f) and (g). Positive results in the two identification tests are required to confirm the presence of the pest.

4. SAMPLE PREPARATION

The standard sample shall contain 200 tubers per test. The appropriate laboratory procedure to process the heel end cores to obtain the extract for detection of the specified pest is described in international diagnostic standards.

Detection of latent infections shall be carried out on composite samples of stem segments or leaf petioles. The procedure may be applied for up to 200 stem parts or 200 leaf petioles from different plants in one sample. The appropriate laboratory procedure to disinfect and process the stem or leaf petiole segments to obtain the extract for the detection of the specified pest is described in international diagnostic standards.

Sections of tissue shall be aseptically removed from the vascular ring in a potato tuber or from the vascular strands in stems of specified plants showing wilting symptoms. The appropriate laboratory procedure to process these tissues to obtain the extract for the detection of the specified pest is described in full detail in international diagnostic standards.

The principal test for the detection of the specified pest in samples of surface water, water from recirculation systems and effluent samples (potato processing industry) is selective isolation. The appropriate laboratory procedure to process water samples is described in international diagnostic standards.

ANNEX II

Template for presenting the results of the brown rot surveys carried out during the calendar year preceding the year of reporting