Anthoine, G.

Buisson, A.

Sarniguet, C.

English

JKI Open Journal Systems (Julius Kühn-Institut)

  IUFRO / REPHRAME International Conference on Pine Wilt Disease 2013  Berichte aus dem Julius Kühn-Institut 169. 2013 96 Sarniguet C et al., Bursaphelenchus xylophilus identification, from literature to routine analysis: how to make morphological analysis reliable. In: Schröder, T. (ed.), Pine Wilt Disease Conference 2013, pp. 96-97, Braunschweig, ISSN: 1866-590X (101) Bursaphelenchus xylophilus identification, from literature to routine analysis: how to make morphological analysis reliable. Sarniguet CLaboratoire de la santé des végétaux, Unité de nématologie, Domaine de la Motte, BP35329, F-35653 Le Rheu cedex, France , Buisson A, Anthoine G   Email: corinne.sarniguet@anses.fr  Official surveys on Bursaphelenchus xylophilus are conducted in a compulsory manner since 2000 in European Union including in France. Considering the Portuguese situation and the dispersal of Bursaphelenchus xylophilus throughout its territory and recent occurrences in Spain, the reliability of sampling and analysis undertaken is a critical issue to early detect any further dispersal. The implementation of quality assurance for official analysis also led to evaluate the reliability of identification methods, based on morphology or on molecular principles. Published morphological identification keys to xylophilus group (Braasch et al. 2009) and to B. xylophilus species levels (EPPO 2009) were considered for evaluation. The keys were submitted to a panel of slides including B. xylophilus individuals, males and females. Conclusions were drawn about the possibility to observe specific criteria, such as lateral lines number, shape of spicules, number of caudal papillae and presence of vulval flap for the group level (Table 1), and the position of excretory pore for species level (Table 2). Some criteria were shown not to be reliable for routine use: number of caudal papillae and excretory pore position. Consequently, reliable identification keys for B. xylophilus were designed (Table 3) and validated according to a standardized process and taking into account recommendations from EPPO protocol (EPPO 2010). Additional results are available in Sarniguet et al. (2013). REFERENCES Braasch H; Burgermeister W; Gu J (2009). Revised intra-generic grouping of Bursaphelenchus  Fuchs, 1937 (Nematoda: Aphelenchoididae). Journal of Nematode Morphology and Systematics 12 (1), 65-88.   IUFRO / REPHRAME International Conference on Pine Wilt Disease 2013  Berichte aus dem Julius Kühn-Institut 169. 2013 97 EPPO (2009). EPPO standards PM7/4 (2) Diagnostics. Bursaphelenchus xylophilus. Bulletin OEPP/EPPO bulletin 39, 344-353. EPPO (2010). EPPO standards PM7/98 (1) Diagnostics. Specifics requirements for laboratories preparing accreditation for plant pest diagnostic activity. Bulletin OEPP/EPPO bulletin 40, 5-22. Sarniguet C; Buisson A; Anthoine G (2013). Validation of morphological keys for identification of Bursaphelenchus xylophilus (Nematoda, Parasitaphelenchidae) to group and species level. Bulletion OEPP/EPPO bulletin 43 (2), 255-261. Table 1. Results of evaluation of the xylophilus group key according to Braasch et al. (2009): number of conform observation/total number of observations Observed criteria Females Males Lateral lines 23/60 41/60 Spicules NA 60/60 Caudal papillae NA 0/60 Vulval flap 46/60 NA NA: not applicable  Table 2.  Results of evaluation of the EPPO (2009) B. xylophilus identification key: number of times the criteria is observed/ total number of observations Features to be observed Results Excretory pore not observed 29/60 Excretory pore not at expected place 11/60 Position of the excretory pore conform to B. xylophilus 20/60  Table 3.  Key designed for Bursaphelenchus xylophilus species identification from female individuals  1 Female with conical or slender tail with or without mucro   not B. xylophilus  Female with sub-cylindrical tail ………………………… 2 2 Female with sub-cylindrical tail, rounded end without mucro   B. xylophilus  Female with sub-cylindrical tail with a terminal mucro   not B. xylophilus or B. xylophilus mucronate form (1) (1) molecular identification needed 

Bursaphelenchus xylophilus identification, from literature to routine analysis: how to make morphological analysis reliable

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Bursaphelenchus xylophilus identification, from literature to routine analysis: how to make morphological analysis reliable

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