HomePlant DiseaseVol. 105, No. 3First Report of Three Pineapple Mealybug Wilt-Associated Viruses in Queen Victoria Pineapples in Reunion Island PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report of Three Pineapple Mealybug Wilt-Associated Viruses in Queen Victoria Pineapples in Reunion IslandD. Massé, N. Cassam, B. Hostachy, M.-L. Iskra-Caruana, M. Darnaudery, P. Lefeuvre, and J.-M. LettD. MasséANSES, Laboratoire de la santé des végétaux (LSV), Unité RAPT, F-97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this author, N. CassamANSES, Laboratoire de la santé des végétaux (LSV), Unité RAPT, F-97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this author, B. HostachyANSES, Laboratoire de la santé des végétaux (LSV), Unité RAPT, F-97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this author, M.-L. Iskra-CaruanaCIRAD, UMR BGPI, F-34398 Montpellier, FranceBGPI, Université de Montpellier, CIRAD, INRAE, Institut Agro, Montpellier, FranceSearch for more papers by this author, M. DarnauderyCIRAD, UPR HortSys, F-97455 Saint-Pierre, La Réunion, FranceHortSys, Université de Montpellier, CIRAD, Montpellier, FranceSearch for more papers by this author, P. LefeuvreCIRAD, UMR PVBMT, Pôle de Protection des Plantes, F-97410 Saint-Pierre, La Réunion, FrancePVBMT, Université de La Réunion, CIRAD, Saint-Pierre, FranceSearch for more papers by this author, and J.-M. Lett†Corresponding author: J.-M. Lett; E-mail Address: lett@cirad.frhttp://orcid.org/0000-0003-1585-5338CIRAD, UMR PVBMT, Pôle de Protection des Plantes, F-97410 Saint-Pierre, La Réunion, FrancePVBMT, Université de La Réunion, CIRAD, Saint-Pierre, FranceSearch for more papers by this author AffiliationsAuthors and Affiliations D. Massé1 N. Cassam1 B. Hostachy1 M.-L. Iskra-Caruana2 3 M. Darnaudery4 5 P. Lefeuvre6 7 J.-M. Lett6 7 † 1ANSES, Laboratoire de la santé des végétaux (LSV), Unité RAPT, F-97410 Saint-Pierre, La Réunion, France 2CIRAD, UMR BGPI, F-34398 Montpellier, France 3BGPI, Université de Montpellier, CIRAD, INRAE, Institut Agro, Montpellier, France 4CIRAD, UPR HortSys, F-97455 Saint-Pierre, La Réunion, France 5HortSys, Université de Montpellier, CIRAD, Montpellier, France 6CIRAD, UMR PVBMT, Pôle de Protection des Plantes, F-97410 Saint-Pierre, La Réunion, France 7PVBMT, Université de La Réunion, CIRAD, Saint-Pierre, France Published Online:13 Jan 2021https://doi.org/10.1094/PDIS-05-20-1068-PDNAboutSectionsView articlePDFPDF Plus ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat View articleMealybug wilt of pineapple is one of the most destructive diseases of pineapple (Ananas comosus) in the world (Sether et al. 2005). Viruses in three distinct species of the genus Ampelovirus (family Closteroviridae), Pineapple mealybug wilt-associated virus-1, virus-2, and virus-3, have been identified in several pineapple-growing regions worldwide such as Hawaii (Hu et al. 1993), Australia (Wakman et al. 1995), Central and South America (Borroto et al. 1998), Ecuador (Alvarez et al. 2015), and recently West Africa (Nyarko and Asare-Bediako 2019). The ‘Queen Victoria’ cultivar is the most widely cultivated pineapple in Reunion Island and is the main fruit crop exported. From October 2016 to February 2018, leaves from four pineapples (cv. Queen Victoria) from four different plots showing symptoms of wilt disease were collected in Reunion Island. Three sets of primers were used in reverse transcription polymerase chain reaction (RT-PCR) for the specific detection of members of each virus species: PMW1dF/R for pineapple mealybug wilt-associated virus-1 (PMWaV-1) (Gambley et al. 2009), PMWaV2-223/224 for PMWaV-2 (Sether et al. 2005), and Wilt3dF/R for PMWaV-3 (Gambley et al. 2009). Primer sets were designed from the RdRp genes for PMWaV-1 and -3, and from the HsP70 gene for PMWaV-2. Expected DNA fragments of 303, 610, and 424 nucleotides (nt) were obtained from the four samples for PMWaV-1 and -2, and from one sample for PMWaV-3, respectively. The fragments were directly sequenced in both directions, assembled, and analyzed (Geneious version R11.1.2). The four consensus sequences (accession nos. MT447832 to 835), obtained from the 303-nt DNA fragments, shared 91.6 to 95.8% nt and 95.5 to 97% amino acid (aa) identities with Hawaiian (AF414119, MH704740) and Australian (EF467924, EF467925, and EF463006) isolates of PMWaV-1. The four consensus sequences (MT469951 to 954), obtained from the 610-nt DNA fragments, shared 98.3 to 99.8% nt and 97.7 to 100% aa identities with Hawaiian isolates of PMWaV-2 (AF283103, MH704741). The single consensus sequence (MT469955), obtained from the 424-nt DNA fragments, shared 96 to 97.3% nt and 100% aa identities with Hawaiian (DQ399259, MH704742) and Australian (EF467918) isolates of PMWaV-3. To further confirm the presence of members of PMWaV-1, -2, and -3 in the four pineapple samples, three specific primer sets designed from the coat protein (CP) genes were used (FJ08-1/2 [Shen et al. 2009] CP229/CP230, and CP231/CP232 [Hernandez-Rodriguez et al. 2014], respectively, for each virus). Amplicons of the expected sizes were obtained from the four samples for PMWaV-2, and from two samples for PMWaV-1 and -3. The direct sequencing of the amplicons confirmed the previous results. The consensus sequences of the CP of PMWaV-1 (MT990947 to 948), PMWaV-2 (MT990949 to 952), and PMWaV-3 (MT990953 to 954) shared 98 to 100% nt and 99 to 100% aa identities with several isolates of the viruses from Hawaii (MH704740 to 742, MN539274), Taiwan (LC507819), and Cuba (DQ225114). To our knowledge, this is the first report of PMWaV-1, -2, and -3 on wilt diseased pineapples in Reunion Island. The spread of this ampelovirus complex in the pineapple fields of Reunion represents potentially a major threat for this agricultural export sector.The author(s) declare no conflict of interest.References:Alvarez, R. A., et al. 2015. New Dis. Rep. 31:15. https://doi.org/10.5197/j.2044-0588.2015.031.015 Crossref, Google ScholarBorroto, E. G., et al. 1998. Plant Dis. 82:263. https://doi.org/10.1094/PDIS.1998.82.2.263C Link, ISI, Google ScholarGambley, C. F., et al. 2009. J. Virol. Methods 155:187. https://doi.org/10.1016/j.jviromet.2008.10.006 Crossref, ISI, Google ScholarHernandez-Rodriguez, L., et al. 2014. Crop Prot. 65:43. https://doi.org/10.1016/j.cropro.2014.07.003 Crossref, ISI, Google ScholarHu, J. S., et al. 1993. Acta Hortic. 334:411. https://doi.org/10.17660/ActaHortic.1993.334.44 Crossref, Google ScholarNyarko, J., and Asare-Bediako, E. 2019. New Dis. Rep. 40:18. https://doi.org/10.5197/j.2044-0588.2019.040.018 Crossref, Google ScholarSether, D. M., et al. 2005. Plant Dis. 89:450. https://doi.org/10.1094/PD-89-0450 Link, ISI, Google ScholarShen, B. N., et al. 2009. Plant Dis. 93:196. https://doi.org/10.1094/PDIS-93-2-0196C Link, ISI, Google ScholarWakman, W., et al. 1995. Aust. J. Agric. Res. 46:947. https://doi.org/10.1071/AR9950947 Crossref, ISI, Google ScholarThe author(s) declare no conflict of interest.Funding: Funding was provided by European Union - Conseil Régional de La Réunion (GURDT / 12016-1731-0006632).DetailsFiguresLiterature CitedRelated Vol. 105, No. 3 March 2021SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionMaize ear showing scattered moldy or “starburst” symptoms, typical of Fusarium verticillioides (A. F. Logrieco et al.). Photo credit: G. Munkvold. Bright yellow vein banding, rings, and lines associated with alfalfa mosaic virus infection in chayote (Sechium edule (Jacq.) Sw.) (G. Parrella et al.). Photo credit: G. Parrella. Metrics Downloaded 491 times Article History Issue Date: 26 Mar 2021Published: 13 Jan 2021Accepted: 7 Oct 2020 Pages: 715-715 Information© 2021 The American Phytopathological SocietyFundingEuropean Union - Conseil Régional de La RéunionGrant/Award Number: GURDT / 12016-1731-0006632Keywordswilt diseaseAnanas comosusAmpelovirusSouth West Indian Ocean IslandsThe author(s) declare no conflict of interest.

HomePlant DiseaseVol. 103, No. 11First Report of Orange Rust Caused by Puccinia kuehnii on Sugarcane on the Island of Reunion PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report of Orange Rust Caused by Puccinia kuehnii on Sugarcane on the Island of ReunionJ. Hubert, C. Jeandel, L. Costet, B. Hostachy, A.-S. Dupuis, A. Coddeville, L. Barau, and R. IoosJ. HubertANSES Laboratoire de la Santé des Végétaux, Unité de Mycologie, 54220 Malzéville, FranceSearch for more papers by this author, C. JeandelANSES Laboratoire de la Santé des Végétaux, Unité de Mycologie, 54220 Malzéville, FranceSearch for more papers by this author, L. CostetCIRAD, UMR PVBMT, Saint-Pierre, La Réunion, FranceSearch for more papers by this author, B. HostachyANSES Laboratoire de la Santé des Végétaux, Unité des Ravageurs et Agents Pathogènes Tropicaux, Saint-Pierre, La Réunion, FranceSearch for more papers by this author, A.-S. DupuisDAAF 974, Service de l’Alimentation, La Réunion, FranceSearch for more papers by this author, A. CoddevilleDAAF 974, Service de l’Alimentation, La Réunion, FranceSearch for more papers by this author, L. BarauERCANE, 97490 Saint-Denis, La Réunion, FranceSearch for more papers by this author, and R. Ioos†Corresponding author: R. Ioos; E-mail Address: [email protected]http://orcid.org/0000-0001-9359-5098ANSES Laboratoire de la Santé des Végétaux, Unité de Mycologie, 54220 Malzéville, FranceSearch for more papers by this authorAffiliationsAuthors and Affiliations J. Hubert1 C. Jeandel1 L. Costet2 B. Hostachy3 A.-S. Dupuis4 A. Coddeville4 L. Barau5 R. Ioos1 † 1ANSES Laboratoire de la Santé des Végétaux, Unité de Mycologie, 54220 Malzéville, France 2CIRAD, UMR PVBMT, Saint-Pierre, La Réunion, France 3ANSES Laboratoire de la Santé des Végétaux, Unité des Ravageurs et Agents Pathogènes Tropicaux, Saint-Pierre, La Réunion, France 4DAAF 974, Service de l’Alimentation, La Réunion, France 5ERCANE, 97490 Saint-Denis, La Réunion, France Published Online:28 Aug 2019https://doi.org/10.1094/PDIS-04-19-0750-PDNAboutSections ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Puccinia kuehnii (W. Krüger) E. J. Butler, the causal agent of orange rust, is reported as one of the two known widely spread rust species on commercial sugarcane (Saccharum spp.). This pathogen affects plant foliage, and the symptoms are characterized by the appearance of small elongated uredinial lesions (pustules), typically light orange to yellowish-brown, oval in shape, producing urediniospores. On highly susceptible cultivars, abundant pustules develop and coalesce on the lower leaf surface, causing tissue death. On such cultivars, orange rust has severe yield and economic impacts (Magarey et al. 2001). This pathogen has expanded its range of distribution over the last 10 years and is now affecting America and Africa. In July and August 2018, in different areas of Reunion Island, typical rust symptoms were observed on leaves of R062006 and R062007 sugarcane varieties in selection. The intensity of the disease was low to moderate. Samples were collected on both varieties in fields from the ERCANE breeding station in Saint-Louis (Le Gol) in the southwest (–21.265027; 55.379830) and in Saint-Philippe (Le Baril) in the southeast (–21.359220; 55.731959) for laboratory analysis. The lesions were examined under a dissecting microscope. Microscopic slides were prepared from material by carefully picking up spores in rust sori with a needle. The presence of the causal fungus of orange rust was confirmed by the observation of typical microscopic features. Urediniospores were mostly obovoid to pyriform, echinulate, variable in size, 35.9 to 57.1 × 16.4 to 29.5 µm. Walls were 1 to 2.5 µm thick, most often with apical thickening up to 8.3 µm. A few inconspicuous hyaline paraphyses with a thin wall (1 to 2 µm) were present. No teliospore could be observed (Dixon et al. 2010; Virtudazo et al. 2001). Identity of the species involved was determined by sequencing the internal transcribed spacer (ITS) region of the rDNA followed by comparison with reference sequences available in GenBank. Leaf material containing uredinial lesions was cut into pieces about 2 × 3 mm and transferred into 2-ml microtubes. Total DNA was then extracted using a commercial plant DNA extraction kit (DNeasy Plant Mini Kit, Qiagen), and the fungal ITS region was amplified by PCR using specific primers PkPm-F/R, located respectively in ITS1 and ITS2 (Glynn et al. 2010). The nucleotide sequence was determined and deposited on GenBank (MK578656 to MK578657). Analysis of the sequence by BLAST showed 100 and 99% identity with P. kuehnii, over a 494- and 493-bp length, respectively, which was consistent with the morphological features observed. This is the first report of P. kuehnii in Reunion Island. Up to now, the disease has only been found in cultivar trials in Reunion. Orange rust has spread rapidly in the sister Mascarene Island of Mauritius since it was initially detected in March 2018. Out of four cultivars present in Mauritius and planted on Reunion, three tend to be moderately or slightly susceptible, and one is symptomless (S. Saumtally, personal communication). Following the emergence of brown rust caused by Puccinia melanocephala Syd. & P. Syd. in Reunion in 1965 (Boyer de la Giroday et al. 1979), resistant cultivars, such as R570 bearing the BruI resistance gene (Costet et al. 2012), have been bred and planted to control this rust species. The same work will have to be conducted by the breeders in the years to come in order to take into account this new threat.The author(s) declare no conflict of interest.References:Boyer de la Giroday, E., et al. 1979. Agron. Trop. 34:372. Google ScholarCostet, L., et al. 2012. Theor. Appl. Genet. 125:825. https://doi.org/10.1007/s00122-012-1875-x Crossref, ISI, Google ScholarDixon, L. J., et al. 2010. Mycol. Prog. 9:459https://doi.org/10.1007/s11557-009-0649-6 Crossref, ISI, Google ScholarGlynn, N. C., et al. 2010. Plant Pathol. 59:703. https://doi.org/10.1111/j.1365-3059.2010.02299.x Crossref, ISI, Google ScholarMagarey, R. C., et al. 2001. Proc. Int. Soc. Sugar Cane Technol. 24:410. Google ScholarVirtudazo, E. V., et al. 2001. Mycoscience 42:167. https://doi.org/10.1007/BF02464133 Crossref, Google ScholarThe author(s) declare no conflict of interest.DetailsFiguresLiterature CitedRelated Vol. 103, No. 11 November 2019SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionAdvanced symptoms of bacterial blotch disease on mushroom caps (Osdaghi et al.). Photo credit: C. Bull. Powdery mildew caused by Golovinomyces neosalviae on Salvia fruticosa (Soylu et al.). Photo credit: S. Soylu. Metrics Article History Issue Date: 4 Nov 2019Published: 28 Aug 2019First Look: 25 Jun 2019Accepted: 17 Jun 2019 Page: 2962 InformationThis article is in the public domain and not copyrightable. It may be freely reprinted with customary crediting of the source The American Phytopathological Society, 2019.Keywordsfungitropical plantspathogen detectionThe author(s) declare no conflict of interest.Cited bySugarcane Breeding in Reunion: Challenges, Achievements and Future Prospects17 May 2021 | Sugar Tech, Vol. 24, No. 1Puccinia kuehnii (orange rust)CABI Compendium, Vol. CABI CompendiumOrange Rust Disease Progress in Different Plantation Cycles of Sugarcane in Cuba3 July 2020 | Sugar Tech, Vol. 22, No. 6

Pseudocercospora fijiensis, the causal agent of black Sigatoka disease of banana (Musa spp.) (BLSD), is considered to be the major economic threat for banana cultivated for export (de Lapeyre de Bellaire et al., 2010). The disease has a …

Pseudocercospora fijiensis, the causal agent of black Sigatoka disease of banana (Musa spp.) (BLSD), is considered to be the major economic threat for banana cultivated for export (de Lapeyre de Bellaire et al., 2010). The disease has a …

New Disease ReportsVolume 36, Issue 1 p. 19-19 ArticleOpen Access Occurrence of cassava brown streak disease and associated Cassava brown streak virus and Ugandan cassava brown streak virus in the Comoros Islands H.A. Azali, H.A. Azali INRAPE, ex-CEFADER, Mde Ngazidja, Moroni, Union des Comores Université des Comores, Rue de la Corniche, Moroni, Union des ComoresSearch for more papers by this authorV. Maillot, V. Maillot CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorN. Cassam, N. Cassam ANSES-LSV Réunion, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorT. Chesneau, T. Chesneau CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, France Lycée Agricole Professionnel de Coconi, 97670 Coconi, Mayotte, FranceSearch for more papers by this authorJ. Soulezelle, J. Soulezelle CIRAD, UMR PVBMT, Station Agronomique de Dembeni, 97660 Dembeni, Mayotte, FranceSearch for more papers by this authorS. Scussel, S. Scussel CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorA.L. Abdoul Karime, A.L. Abdoul Karime DAAF Mayotte, Service de l'Alimentation – Santé des Végétaux, 97600 Mamoudzou, Mayotte, FranceSearch for more papers by this authorB. Hostachy, B. Hostachy ANSES-LSV Réunion, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorB. Reynaud, B. Reynaud CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorM. Roux-Cuvelier, M. Roux-Cuvelier CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorI. Robène, I. Robène CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorJ.-M. Lett, Corresponding Author J.-M. Lett lett@cirad.fr CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this author H.A. Azali, H.A. Azali INRAPE, ex-CEFADER, Mde Ngazidja, Moroni, Union des Comores Université des Comores, Rue de la Corniche, Moroni, Union des ComoresSearch for more papers by this authorV. Maillot, V. Maillot CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorN. Cassam, N. Cassam ANSES-LSV Réunion, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorT. Chesneau, T. Chesneau CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, France Lycée Agricole Professionnel de Coconi, 97670 Coconi, Mayotte, FranceSearch for more papers by this authorJ. Soulezelle, J. Soulezelle CIRAD, UMR PVBMT, Station Agronomique de Dembeni, 97660 Dembeni, Mayotte, FranceSearch for more papers by this authorS. Scussel, S. Scussel CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorA.L. Abdoul Karime, A.L. Abdoul Karime DAAF Mayotte, Service de l'Alimentation – Santé des Végétaux, 97600 Mamoudzou, Mayotte, FranceSearch for more papers by this authorB. Hostachy, B. Hostachy ANSES-LSV Réunion, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorB. Reynaud, B. Reynaud CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorM. Roux-Cuvelier, M. Roux-Cuvelier CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorI. Robène, I. Robène CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this authorJ.-M. Lett, Corresponding Author J.-M. Lett lett@cirad.fr CIRAD, UMR PVBMT, Pôle de Protection des Plantes, 97410 Saint-Pierre, La Réunion, FranceSearch for more papers by this author First published: 25 November 2017 https://doi.org/10.5197/j.2044-0588.2017.036.019Citations: 3AboutSectionsPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Cassava brown streak disease (CBSD) has emerged as the most important viral disease of cassava (Manihot esculenta) in Africa and is a major threat to food security. The expanding CBSD epidemic in East and Central Africa is caused by two ipomoviruses (family Potyviridae), Cassava brown streak virus (CBSV; Monger et al., 2) and Ugandan cassava brown streak virus (UCBSV; Winter et al., 5). In 2014, symptoms of yellowing on leaves and brown necrotic rot on tuberous roots on different cassava landraces on the west coast of Mayotte (Maoré) were associated with CBSD and UCBSV (Roux-Cuvelier et al., 2014). Similar symptoms were observed in April 2015 in the north of Mayotte and in July and September 2016 in the three other islands of the Comoros archipelago: Grande Comore (Ngazidja), Anjouan (Ndzuwani) and Mohéli (Mwali) (Figs. 1). To evaluate the disease epidemic in the south-western Indian Ocean islands and to verify the identity of the causal agents, symptomatic leaf samples of 67 affected plants were collected and tested. Figure 1Open in figure viewerPowerPoint Figure 2Open in figure viewerPowerPoint Total RNA was extracted from these samples using the RNeasy Plant Mini Kit (Qiagen, France). For the detection of cassava brown streak viruses, two-step RT-PCR was performed using the CBSDDF2 and CBSDDR primers (Mbanzibwa et al., 1). These primers were designed for simultaneous specific detection of CBSV and UCBSV (amplicons of 344 and 440 bp, respectively). RT-PCR products of the expected sizes for CBSV and UCBSV were obtained for 31 and 14 samples, respectively (Table 1). To confirm the diagnosis, the RT-PCR products were directly and bi-directionally sequenced and contigs assembled using DNA Baser (Heracle BioSoft, Romania). Thirty-seven nucleotide sequences were obtained from the 45 RT-PCR positive samples (GenBank Accession Nos. MF948187-MF948223; Table 1). Twelve sequences of 374 bp showed the highest nucleotide identity (91-93%) with an isolate of UCBSV from Mayotte (LN611675). The other 25 sequences of 280 bp showed the highest nucleotide identity (96-99%) with isolates of CBSV from Uganda, Kenya and Tanzania (KJ606250, LT560330 and GU563322). A phylogenetic reconstruction with publicly available complete genome sequences of cassava brown streak viruses confirmed that CBSV and UCBSV isolates from Grande Comore, Anjouan, Mohéli and Mayotte are genetically related to isolates from East Africa (MEGA6; Tamara et al., 4; Fig. 3). Altogether, these results demonstrate that CBSD epidemic has reached the four islands of the Comoros archipelago and as in East and Central Africa the disease is caused by two ipomoviruses, CBSV and UCBSV. Figure 3Open in figure viewerPowerPoint Table 1. Geographical origin of cassava samples presenting cassava brown streak symptoms and identification of Cassava brown streak virus (CBSY) and Ugandan cassava brown streak virus (UCBSV) isolates. Reference Sampling date Locality Island Amplicon size (bp) RT-PCR diasnosis Sequencing result; GenBank Accession No. MAB1 15/09/2016 Mababani Grande Comore 334 CBSV Not available MAB2 15/09/2016 Mababani Grande Comore 440 UCBSV Not available HEL1A 15/09/2016 Helindje Grande Comore 440 UCBSV UCBSV; MF94S194 HEL IB 15/09/2016 Helindje Grande Comore 440 UCBSV UCBSV; MF94S195 HEL2B 15/09/2016 Helindje Grande Comore 440 UCBSV UCBSV; MF94S196 HELSA 15/09/2016 Helindje Grande Comore 440 UCBSV UCBSV; MF94S197 HEL3B 15/09/2016 Helindje Grande Comore 440 UCBSV UCBSV; MF94S19S OUZ2A 15/09/2016 Ouzio Grande Comore 334 CBSV CBSV; MF94S217 OUZ2B 15/09/2016 Ouzio Grande Comore 334 CBSV CBSV; MF94S21S 0OUZ2C 15/09/2016 Ouzio Grande Comore 334 CBSV CBSV; MF94S219 MAKI A 15/09/2016 Makcrani Grande Comore 334 CBSV CBSV; MF94S20S KOPIE 15/09/2016 Kopve Grande Comore 334 CBSV Not available NTS1A 15/09/2016 Nts immoicbongo Grande Comore 334 CBSV CBSV; MF94S211 NTS 1B 15/09/2016 Nts immoicbongo Grande Comore 334 CBSV CBSV; MF94S212 NTS 1C 15/09/2016 Nts immoicbongo Grande Comore 334 CBSV CBSV: MF94S213 NTS ID 15/09/2016 Nts immoicbongo Grande Comore 334 CBSV CBSV; MF94S214 NTS2A 15/09/2016 Nts immoicbongo Grande Comore 334 CBSV CBSV; MF94S215 NDEl 15/09/2016 Ndemani Grande Comore 440 UCBSV Not available NDE2A 15092016 Ndemani Grande Comore 334 CBSV Not available NDE2B 15/09/2016 Ndemani Grande Comore 334 CBSV CBSV; MF94S209 NDE2C 15/09/2016 Ndemani Grande Comore 334 CBSV CBSV; MF94S210 OUZ1 15092016 Ouzio Grande Comore 334 CBSV CBSV; MF94S216 HAN1 15/09/2016 Hankobo Grande Comore 440 UCBSV UCBSV; MF94S1S9 HAN2 15/09/2016 Hankobo Grande Comore 440 UCBSV UCBSV; MF94S190 HANS 15092016 Hankobo Grande Comore 440 UCBSV UCBSV; MF94S191 HAN4 15/09/2016 Hankobo Grande Comore 440 UCBSV UCBSV; MF94S192 HAN5 15/09/2016 Hankobo Grande Comore 440 UCBSV UCBSV; MF94S193 CHAI 15092016 Chamle Grande Comore 440 UCBSV UCBSV; MF94S1S7 CHA2 15/09/2016 Cbamle Grande Comore 334 CBSV CBSV; MF94S199 CHAS 15/09/2016 Chamle Grande Comore 334 CBSV CBSV; MF94S200 CHA4 15/09/2016 Chamle Grande Comore 334 CBSV CBSV; MF94S201 CHAS 15/09/2016 Chamle Grande Comore 334 CBSV Not available CHA6 15/09/2016 Chamle Grande Comore 440 UCBSV UCBSV; MF94S1SS HAYA1 17/07/2016 Hayrara Mobeli 334 CBSV CBSV; MF94S206 HAY 1C 17/07/2016 Havrara Moheli 334 CBSV CBSV; MF94S207 SINl 16/07/2016 Not available Mobeli 334 CBSV CBSV; MF94S220 DOM2A 12/07/2016 Domoni Anjouan 334 CBSV CBSV; MF94S203 DOM2B 12/07/2016 Domoni Anjouan 334 CBSV CBSV; MF94S204 DOM2C 12/07/2016 Domoni Anjouan 334 CBSV CBSV; MF94S205 DOM1 12/07/2016 Domoni Anjouan 334 CBSV CBSV; MF94S202 BMM1 12/07/2016 Not available Anjouan 334 CBSV Not available 9B 23/04/2015 Longoni Mayotte 334 CBSV CBSVMF94S221 11B 23/04/2015 Longoni Mayotte 334 CBSV Not available I8A 23/04/2015 Longoni Marotte 334 CBSV CBSV; MF94S222 38B 23/04/2015 Longoni Mayotte 334 C3SV CBSV; MF94S223 This is the first report of CBSD in Grande Comore, Anjouan and Mohéli which demonstrate the expanding epidemic of CBSD in the south-western Indian Ocean islands. Given that cassava is an important staple food in the Comoros archipelago and Madagascar, this report is of great significance for managing the health of cassava in this region, and warrants urgent attention from regulatory institutions. Acknowledgments This work was co-funded by the European Union (ERDF, program INTERREG V), by the Conseil Régional de la Réunion and by CIRAD. HA was supported by a postdoctoral fellowship from the French Embassy in Moroni (Comoros Union). References 1Mbanzibwa DR, Tian YP, Tugume AK, Patil BL, Yadav JS, Bagewadi B, Abarshi MM, Alicai T, Changadeya W, Mkumbira J, Muli MB, Mukasa SB, Tairo F, Baguma Y, Kyamanywa S, Kullaya A, Maruthi MN, Fauquet CM, Valkonen JPT, 2011. Evolution of cassava brown streak disease-associated viruses. Journal of General Virology 92, 974– 987. 10.1099/Vir.0.026922-0 2Monger WA, Alicai T, Ndunguru J, Kinyua ZM, Potts M, Reeder RH, Miano DW, Adams IP, Boonham N, Glover RH, Smith J, 2010. The complete genome sequence of the Tanzanian strain of Cassava brown streak virus and comparison with the Ugandan strain sequence. Archives of Virology 155, 429– 433. 10.1007/s00705-009-0581-8 3Roux-Cuvelier M, Teyssedre D, Chesneau T, Jeffray C, Massé D, Jade K, Abdoul-Karime AL, Hostachy B, Reynaud B, Legg JP(2015). First report of cassava brown streak disease and associated Ugandan cassava brown streak virus in Mayotte Island. New Disease Reports 30, 28. 10.5197/j.2044-0588.2014.030.028 4Tamura K, Stecher G, Peterson D, Filipski A, Kumar S, 2013. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution 30, 2725– 2729. 10.1093/molbev/mst197 5Winter S, Koerbler M, Stein B, Pietruszka A, Paape M, Butgereitt A, 2010. Analysis of cassava brown streak viruses reveals the presence of distinct virus species causing cassava brown streak disease in East Africa. Journal of General Virology 91, 1365– 1372. 10.1099/Vir.0.014688-0 Citing Literature Volume36, Issue1July 2017-December 2017Pages 19-19 FiguresReferencesRelatedInformation

Xanthomonas axonopodis pv. allii (Xaa) is the causal agent of bacterial blight of onion (BBO), an emerging disease threatening the world onion production, and causing damage to other Alliaceae, including garlic, welsh onion, shallot, chive and leek. Xaa is an EPPO quarantine organism (A1 list i.e. absent from the EPPO region). The international spread of Xaa can probably be explained by its seedborne status (Roumagnac et al., 2000). Consequently it is of crucial importance to validate efficient detection methods which allow the detection of seedborne pathogens for which the level of inoculum can be very low in very large seed lots. We evaluate a detection scheme based on molecular tests (a triplex quantitative real-time PCR assay (Robene et al., 2015) and a duplex nested end-point PCR assay (Robene-Soustrade et al., 2010)) and isolation. The method assessment was performed following the European standard EN ISO 16140 and the EPPO standard PM7/98 (2). An intra-laboratory study was first conducted, where we characterized the analytical specificity (inclusivity and exclusivity), the analytical sensitivity and the repeatability. We finally tested the detection scheme on naturally contaminated onion seed lots showing different contamination rates. In addition to the intralaboratory study, an inter-laboratory reproducibility trial (including five different laboratories) was performed. This detection scheme was shown to be very efficient, combining a maximal inclusivity (100%), a good exclusivity (83%), a high analytical sensitivity (approximately 103 CFU.mL-1), an excellent repeatability (100% for contamination rates higher than 103 CFU.mL-1) and reproducibility (100% for contamination rates higher than 103 CFU.mL-1). These results allowed to validate the detection scheme as official method of analysis for the French officially approved laboratories (MA 038), and to propose it as an EPPO diagnostic protocol for the detection of this emerging pathogen. (Texte integral)

Efficient control of Xanthomonas axonopodis pv. dieffenbachiae, the causal agent of anthurium bacterial blight, requires sensitive and reliable diagnostic tools. The European standard EN ISO 16140:2003 has been followed to compare a nested PCR assay (N-PCR) to a reference method (isolation and serological identification of bacterial colonies) and to other alternative serological detection methods. The evaluation was performed in two steps: a comparative study and a collaborative study involving 15 European laboratories. Although inclusivity was maximal (100%) for all methods, a maximal exclusivity was obtained only with N-PCR followed by an enzymatic restriction digestion of the amplicons. Exclusivity indices of 906, 887 and 472% were found for indirect ELISA, immunofluorescence and double antibody sandwich ELISA, respectively. An exclusivity of 925% was obtained with the reference method, further increased to 100% if pathogenicity tests were performed as a supplemental assay. The best level of sensitivity (relative detection level) was obtained with the reference method followed by the N-PCR assay. The N-PCR performance in terms of relative accuracy, accordance and concordance was very similar to that of the reference method. Moreover, N-PCR had undeniable advantages compared to the reference method (less labour-intensive and less time-consuming). In addition, post-test probabilities of infection were calculated to select the most appropriate detection scheme related to the prevalence of the pathogen. The N-PCR assay has since been included in a revised version of the EPPO detection protocol.

The use of methods of analysis capable of producing reliable analytical results is a prerequisite to the effective control of quarantine plant pathogens. Proficiency testing is considered to be one of the most reliable ways to verify and coordinate analytical results. As a French national reference laboratory in plant pathology, the Anses Plant Health Laboratory organizes proficiency tests in order to ensure that officially approved laboratories (certified by government services) are capable of producing reliable analytical results for the detection of plant pathogens. Proficiency tests in plant pathology have a number of notable features including the processing of qualitative results. This paper presents the experience of the Anses Plant Health Laboratory's Unit for Tropical Pests and Diseases (LSV-RAPT) as an organizer of proficiency tests in plant pathology. The LSV-RAPT has gained recognition for the methodology it has developed in the form of accreditation as a proficiency testing provider according to the ISO/IEC 17043. The methodology can be applied to many other disciplines that use qualitative detection methods.

In November 2013, symptoms of yellowing on leaves and brown necroticrot on tuberous roots (Fig. 1) were observed on different cassava landraces(Manihot esculenta) in Dembeni on Mayotte Island, a French OverseasDepartment in the southwest Indian Ocean. The symptoms, similar to thecassava brown streak disease described in East Africa, suggested thepossible involvement of cassava brown streak viruses (Mbanzibwa et al.,2011). The expanding cassava brown streak disease epidemic in East Africais caused by two ipomoviruses, Cassava brown streak virus (CBSV; Mongeret al., 2010) and Ugandan cassava brown streak virus (UCBSV; Winter etal., 2010). To verify the identity of the causal agent, symptom-bearing leafsamples of 17 affected plants were collected in March and June 2014. Total RNA was extracted from these samples using the RNeasy Plant MiniKit (Qiagen, France). For the detection of cassava brown streak viruses, atwo-step RT-PCR using CBSDDF2 and CBSDDR primers was used(Mbanzibwa et al., 2011). These primers were designed for simultaneousvirus-specific detection of CBSV and UCBSV (amplicons of 344 bp and440 bp, respectively). RT-PCR products of the expected size for UCBSVwere obtained for 11 samples. To confirm the diagnosis, the RT-PCRproducts were directly and bi-directionally sequenced and contigsassembled using DNABaser (Heracle BioSoft, Romania). Sequences of 376bp from eight of the 11 samples were obtained which had 99-100%nucleotide identity (MEGA6; Tamura et al., 2013) (EMBL Accession Nos.LN611671 to LN611678; Table 1). The sequences showed the highestnucleotide identity (86%) with isolates of UCBSV from Uganda andMalawi (FN434109; FN433932) and 85% with an isolate of CBSV fromMozambique (FN434436). A phylogenetic reconstruction with publiclyavailable complete genome sequences of cassava brown streak virusesconfirmed that the Mayotte isolates are genetically more related to UCBSVisolates than to CBSV isolates from East Africa, and represent a separatelineage that appears to be almost intermediate between current isolates ofCBSV and UCBSV (MEGA6; Fig. 2). This is the first report of cassavabrown streak disease in Mayotte. Given that cassava is a staple food in theComoros archipelago and Madagascar, this report is of great significancefor managing the health of cassava in this region, and warrants urgentattention from regulatory institutions.

In November 2013, symptoms of yellowing on leaves and brown necrotic rot on tuberous roots (Fig. 1) were observed on different cassava landraces (Manihot esculenta) in Dembeni on Mayotte Island, a French Overseas Department in the southwest Indian Ocean. The symptoms, similar to the cassava brown streak disease described in East Africa, suggested the possible involvement of cassava brown streak viruses (Mbanzibwa et al., 2011). The expanding cassava brown streak disease epidemic in East Africa is caused by two ipomoviruses, Cassava brown streak virus (CBSV; Monger et al., 2010) and Ugandan cassava brown streak virus (UCBSV; Winter et al., 2010). To verify the identity of the causal agent, symptom-bearing leaf samples of 17 affected plants were collected in March and June 2014. Total RNA was extracted from these samples using the RNeasy Plant Mini Kit (Qiagen, France). For the detection of cassava brown streak viruses, a two-step RT-PCR using CBSDDF2 and CBSDDR primers was used (Mbanzibwa et al., 2011). These primers were designed for simultaneous virus-specific detection of CBSV and UCBSV (amplicons of 344 bp and 440 bp, respectively). RT-PCR products of the expected size for UCBSV were obtained for 11 samples. To confirm the diagnosis, the RT-PCR products were directly and bi-directionally sequenced and contigs assembled using DNABaser (Heracle BioSoft, Romania). Sequences of 376 bp from eight of the 11 samples were obtained which had 99-100% nucleotide identity (MEGA6; Tamura et al., 2013) (EMBL Accession Nos. LN611671 to LN611678; Table 1). The sequences showed the highest nucleotide identity (86%) with isolates of UCBSV from Uganda and Malawi (FN434109; FN433932) and 85% with an isolate of CBSV from Mozambique (FN434436). A phylogenetic reconstruction with publicly available complete genome sequences of cassava brown streak viruses confirmed that the Mayotte isolates are genetically more related to UCBSV isolates than to CBSV isolates from East Africa, and represent a separate lineage that appears to be almost intermediate between current isolates of CBSV and UCBSV (MEGA6; Fig. 2). This is the first report of cassava brown streak disease in Mayotte. Given that cassava is a staple food in the Comoros archipelago and Madagascar, this report is of great significance for managing the health of cassava in this region, and warrants urgent attention from regulatory institutions.

Huanglongbing is an unculturable vascular citrus pathogen transmitted from infected to healthy plants through grafting or by citrus psyllids, Diaphorina citri mainly in Asia and America and Trioza erytreae in Africa. This phloem limited gram-negative bacterium causes dramatic yield losses and is classified into three species based on 16S rDNA sequence analysis (2): (i) ‘Candidatus Liberibacter asiaticus’ (Las), the most epidemiologically active, widespread and heat tolerant species; (ii) ‘Ca. L. africanus’ (Laf), only found in Africa; and (iii) the newly described ‘Ca. L. americanus’ (Lam), which appeared in 2005 in Brazil (5). Considered as a quarantine organism in America and Europe, Las is actively affecting North America and Asia, and research is leading toward psyllid management and resistance breeding. Despite the fact that Reunion Island has successfully controlled Las by introducing a psyllid parasitoid, Tamarixia radiata (1), this strategy was less effective or reproducible within other territories. D. citri was first detected in Guadeloupe in 1998, where the control of the the psyllid population has been effective with T. radiata (3); and was first detected in Martinique in 2012. Following the outbreak in the United States and the Caribbean, and also supported by reports of symptoms in citrus orchards, local National Plant Protection Organizations (NPPO) organized a detection survey across both islands to verify the occurrence of Huanglongbing. Since 2012, 450 sites were prospected each year in Martinique and Guadeloupe, where 20 leaves from 10 to 30 trees were analyzed. DNA extraction was performed (DNeasy Plant Mini Kit, Qiagen) on fresh or dried leaf midribs, along with negative control midribs (Citrus paradisi ‘Star Rubis’) and PCR amplification was done with the species-specific primers A2/J5 (4) and GB1/GB3 (5). Only Las-specific 703-bp amplicons were obtained (n = 43) and 20 were sequenced (Beckman Coulter Genomics, United Kingdom; sequences available through GenBank Accession Nos. KF699074 to KF699093) and blasted against the National Center for Biotechnology Information non-redondant database (NCBI-nr). BLAST analysis revealed 100% identity with the 50S ribosomal protein subunit L1 (rplA) and L10 (rplJ) of ‘Ca. L. asiaticus’ (all strains), and no significant homology to other organisms. Additionally, sequence assembly on a reference genome (NC_012985) showed 100% homology. Huanglongbing was detected in Guadeloupe on March 2012 at Le Moule (East coast) in a Tahiti lime orchard (C. latifolia) and crossed the island in 6 months. Las was detected in Martinique on May 2013 on Tahiti lime (C. latifolia) at Bellefontaine (Northwest) in a private garden and at Le Lorrain (Northeast) in an orchard. Other species from the Rutaceae family were affected by HLB (C. reticulat and C. sinensis) on both islands; however, few of the positive samples showed HLB symptoms (blotchy mottle patterns and green islands on leaves), but presented symptoms similar to nutrient deficiencies. Despite the former presence of T. radiata in Guadeloupe and its detection in Martinique a few weeks after the detection of D. citri, where it had a mean parasitism rate of 70%, an outbreak of HLB spread across both islands. These analyses confirm the presence of HLB in Martinique and Guadeloupe and to our knowledge represent the first report of Las in the French West Indies. Introduction events remain unclear, but this report raises the importance of plant certification, psyllid population control, and surveillance of territories close to the French West Indies, with regards to the risk that HLB presents to citrus production worldwide. References: (1) B. Aubert et al. Fruits. 38, 1983. (2) J. M. Bové. J. Plant Pathol. 88:1, 2006. (3) J. Etienne et al. Fruits. 56:05, 2001. (4) A. Hocquellet et al. Mol. Cell. Probes 13:5, 1999. (5) D. C. Teixeira et al. Mol. Cell. Probes 19:3, 2005.

Asiatic citrus canker disease, caused by Xanthomonas citri pv. citri, seriously impacts citrus production worldwide. Two pathogenic variants, A and A*/Aw, have been described within this pathovar. Two additional pathovars of X. citri with a limited geographic distribution and reduced pathogenicity, namely X. citri pvs. aurantifolii and bilvae, are also pathogenic to citrus and some rutaceous species. Rapid and reliable identification is required for these citrus pathogens, which are classified as a quarantine organism in citrus-producing countries. The specificity of nine polymerase chain reaction primers previously designed for the identification of X. citri pv. citri or citrus bacterial canker strains (both pvs. citri and aurantifolii) was assayed on a large strain collection (n = 87), including the two pathotypes of X. citri pv. citri, other genetic related or unrelated pathogenic xanthomonads, and saprophytic xanthomonads. This study gave congruent results with the original articles when testing the same strains or pathovars but the use of a broad inclusivity and exclusivity panel of strains highlighted new findings. Particularly, primers 2/3, 4/7, and KingF/R failed to provide amplification for three strains from the pathotype A*/Aw. Moreover, all pairs of primers detected at least one non-target strain. These data were supported by in silico analysis of the DNA sequences available from National Center for Biotechnology Information databases.

EFSA guidance regarding pest risk assessment previously identified limitations within a generic qualitative risk assessment scheme published to support EU plant health decision making. In seeking to develop methods to overcome the limitations, we report results of a multiphase project where five test risk assessment methods were applied to Anoplophora glabripennis, Candidatus Phytoplasma pyri, Guignardia citricarpa, Meloidogyne chitwoodi, and Xanthomonas citri strains causing citrus bacterial canker. Teams of pest risk assessors applied the methods to the pests and compared methods in relation to EFSA PLH needs. The most promising method was then revised to incorporate desirable features from the other methods and tested on five more case study pests; Acidovorax citrulli, Ca.P. mali, Ca. P. prunorum, M. Fallax and Mycosphaerella dearnessii. In addition the risk assessment method that emerged from the EU funded project PRATIQUE was adapted to better suit EFSA needs and also tested on the second set of pests. Comprehensive datasheets on each pest were compiled to support the risk assessments. Datasheets included information that emerged from individual pest questionnaires distributed to the National Plant Protection Organisations of each EU Member State. Short term experimental studies were conducted to reduce uncertainty regarding survival and infectivity of the root-knot nematodes in the absence of host plants. Small scale experiments also clarified the role of vectors in the spread of Ca. Phytoplasma prunorum, proving that psyllids acquired the pathogen from wild Prunus sp. and transmitted it to orchards. Results from both sets of studies better informed the respective risk assessments and reduced some uncertainties although significant uncertainties still remain elsewhere within assessments. A novel method to determine an index of individual risk components, e.g. likelihood of entry, was developed. The performance of risk reduction options was evaluated by repeating assessments considering scenarios with and without risk reduction options in place. (Resume d'auteur)

Apres avoir rappele l'histoire des epidemies de TYLCV en culture de tomates a l'ile de la Reunion depuis la detection en 1997 de ce virus et de son vecteur l'aleurode Bemisia tabaci (biotype B), cet article rend compte d'une enquete agronomique menee par le CIRAD et l'ARMEFLHOR en 2010. Elle a etabli ou confirme que : du fait du TYLCV , la filiere tomate a evolue vers la production sous abris (la moitie du tonnage produit) ; la culture de plein champ, compromise a moins de 500 m d'altitude (dans les bas), reste possible dans les Hauts ; sous abris la PB (auxiliaires +insecticides compatible) s'est developpee ; toujours sous abris, les mesures prophylactiques sont indispensables ; partout, l'usage de plants sains (pepinieristes locaux agrees SOC) se developpe ; les varietes resistantes ou tolerantes actuelles sont inadaptees au contexte local. Le maintien de la filiere tomate a la Reunion demande d'ameliorer les techniques de type PBI sous abris, proposer des varietes resistantes ou tolerantes adaptees, tester de nouvelles techniques et mieux connaitre le virus et son vecteur. Les organismes de la filiere et les actions liees a Ecophyto peuvent y aider. La maitrise du TYLCV et son vecteur implique des itineraires techniques associant plusieurs methodes.(resume d'auteur)

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In the frame of a project funded by the European Food Safety Authority (Prima Phacie), effort was put into identifying and testing qualitative plant-pest risk assessment schemes for their suitability in supporting risk management decisions for the European Union. Five schemes were tested, two largely based on the EPPO scheme and three adapted from schemes used in non-European countries. We report the results from the application of these schemes as applied to Xanthomonas citri strains causing Citrus Bacterial Canker, in regard to the risk of its entry, establishment and spread, as well as its potential impact. For this pathogen, three entry pathways into the EU risk assessment area were considered: a) import of fresh citrus fruits, b) import of ornamental rutaceous plants or plant parts, and c) illegal entry of plant propagative material. With the current EU measures in place, of the five schemes tested, two indicated path (c) as that of the highest risk, whereas the other three suggested path (a) as such. This discrepancy is due to the different level of details the components of the risk of entry are considered in each scheme. Most schemes suggested that the establishment potential lay around the mid-range of possible values. All schemes indicated a high rate for potential spread (primarily through human activities) and a medium to high rate for impact potential. The effectiveness of risk management measures was evaluated by comparing results of assessments with and without management measures in place. (Texte integral)