A survey of aphelenchid nematodes (Nematoda: Aphelenchida) associated with maritime pine, Pinus pinaster, was conducted in Portugal in 1996 and 1999. A Bursaphelenchus species has been identified for the first time in the Iberian Peninsula. B. xylophilus is reported for the first time in Europe. It was found in very high numbers - up to 38 000 per 10 g of pine wood - inside a few declining trees infested with curculionid, cerambycid and scolytid beetles. Morphological observations, including shape of spicules, bursa, vulva, female tail end and stylet as well as morphometrics, were in accordance with the species description. Species-specific DNA fragment patterns were obtained using ITS-RFLP analysis, with five different restriction enzymes. The importance and implications of this finding are discussed. Premiere signalisation de Bursaphelenchus xylophilus au Portugal, at en Europe - Une enquete sur les nematodes Aphelenchides (Nematoda: Aphelenchida) associes au pin maritime (Pinus pinaster) a ete realisee au Portugal de 1996 a 1999. Une espece de Bursaphelenchus a ete identifiee pour la premiere fois dans la Peninsule Iberique. B. xylophilus est signale pour la premiere fois en Europe. Il a ete trouve en tres grand nombre - jusqu'a 38 000 individus pour 10 g de bois de pin - dans des arbres deperissants infestes par des Coleopteres Curculionides, Cerambycides et Scolytides. Les observations concernant la morphologie - en particulier la forme des spicules, la bourse, la vulve, l'extremite de la queue de la femelle et le stylet - de meme que les donnees morphometriques correspondent a la description de l'espece. Des sequences de fragments d'ADN specifique de l'espece ont ete obtenus par analyse ITS-RFLP a l'aide de cinq enzymes de restriction. L'importance et les implications de cette decouverte sont discutees.

An analysis of the risk to the countries of the European Union from a possible introduction of Bursaphelenchus xylophilus and its vectors in the genus Monochamus is performed by assembling relevant biological, climatic and commercial information. The risks presented by different trade pathways are assessed and phytosanitary measures to reduce the risks are proposed. The conclusion of the PRA, based on EPPO PRA guideline no. 1, is that these pests present a serious risk to European coniferous forests.

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Abteilung fur nutionale und internationale Angelegenheiten des Pjunzengesundheit, Biologische Bundesunstalt fir Land-und ForstwifischJi, Stuhnsdorfer Dumm 81, 14532 Kleinmuchnow (Germany); e -m i l : h.bruusch@bbu.deThe morphological relationship between European Bursaphelenchus species living in conifers was studied in order to provide key characters for their taxonomic identification. Several species have been newly described or recorded in Europe during the past few years and were morphologically investigated. Among the 28 conifer-inhabiting European species, four groups can be distinguished from each other by the number of lateral incisures, number and position of anal papillae of males and presence and size of a vulval flap of females. Two groups, each containing just one species show two and six lateral lines, respectively, whereas most Bursaphelenchus species belong to two groups, possessing either three or four incisures. Each of the last two groups can be divided into three subgroups, which can be differentiated by spicule shape, number and position of caudal papillae, presence and size of a vulval flap and some other features. A fifth group includes species with unclear group affiliation due to insufficient knowledge of their morphology. The B. xylophilus group (B. xylophilus, B. rnucronatus, B. fraudulentus) with four lateral lines, unique spicule shape and characteristic position of the caudal papillae can clearly be differentiated from all other groups by morphology, also using only light microscopy. Within this group, B. xylophilus can clearly be differentiated morphologically from the other species, provided that adult specimens of both sexes are found and also provided that the round-tailed form of the species remains the only form present, as in Europe. Figures showing the important identification features of spicule shape and female tail shape are given for 28 species. A table shows the complete range of the most important morphometric measurements of these species. Data on the occurrence, host range and, as far as known, the vectors of these species were collected throughout Europe and references are given for each record.

Wood samples were taken from pine tree plantations in different regions of Malaysia and investigated for the occurrence of Bursaphelenchus species. Among 13 samples collected from damaged or dead pines in West Malaysia (near Kuala Lumpur) and Sabah (Kinabalu National Park and Sepilok), only one sample of a dead Pinus caribaea tree with bark beetle attack from a plantation near Kuala Lumpur revealed the presence of a new species of Bursaphelenchus. Bursaphelenchus rainul sp. n. is characterised by a relatively small stylet lacking distinct basal knobs but with slight basal swellings, lateral eld with two lines, female with a small vulval ap, postuterine branch occupying about 33-50% of vulva-anus distance, female tail slim, conoid, with a nely rounded, ventrally bent terminus, male spicules relatively small with high condylus, distinct rostrum but no cucullus, and a small terminal 'bursa' on the male tail. It is similar to B. hellenicus, B. hylobianum and B. abietinus in the shape of the spicules and the female tail as well as in the presence of only two lines in the lateral eld. It is distinguished by a number of characters including spicule size, shape of the 'bursa', female tail shape and excretory pore position. Bursaphelenchus rainul sp. n. is further distinguished from these morphologically similar species by molecular studies using the ITS-RFLP technique.

Bursaphelenchus singaporensis sp. n. isolated in China from packaging wood of deciduous trees, imported from Singapore is described and illustrated. This new species clearly belongs to the B. xylophilus group, having males with the typically shaped spicules with a cucullus at their distal extremity, the typical position and number of caudal papillae (three pairs and one single) and the anterior vulval lip of the females developed as a distinct flap. The new species is characterized by a body length of 792 (553–950) µm and 850 (690–961) µm of males and females, respectively, robust body (a= 34 and 31, resp.), 15–16 µm-long stylet, lateral field with four lines, long postuterine branch (averaging 102 µm) and a strongly conoid female tail (c= 20) with a finely rounded, only slightly ventrally-bent terminus, male with very strong spicules (41–48 µm long), distinct rostrum and small cucullus, and a dorso-ventral visible terminal bursa. Bursaphelenchus singaporensis sp. n. is closely related to other species of the B. xylophilus group (B. xylophilus, B. mucronatus, B. kolymensis, B. fraudulentus, B. conicaudatus, B. baujardi and B. luxuriosae) and similar to B. abruptus. The morphological differentiation is mainly based on the shape of the female tail. However, B. singaporensis sp. n. differs from all other species of the B. xylophilus group by larger spicules. The new species can be differentiated from B. abruptus, B. xylophilus, B. mucronatus, B. fraudulentus, B. conicaudatus and B. luxuriosae by means of ITS-RFLP patterns.

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Wood samples were taken from pine trees in the mountainous region between Pai and Maehongson in northern Thailand. Four species of Bursaphelenchus were recovered from the samples: Bursaphelenchus hylobianum, B. mucronatus, B. aberrans and B. thailandae sp. n. Bursaphelenchus thailandae sp. n. is most similar to B. sychnus but also shows similarities with B. ruehmi, B. hunanensis and B. steineri. It has a relatively small stylet lacking distinct basal knobs and a lateral eld with four lines. The female has more or less protruding vulval lips and a slim, conoid, tail. The relatively small and delicate spicules show a distinctive darker sector and lack a cucullus. The male tail has a very small terminal 'bursa'. Bursaphelenchus thailandae sp. n. is tentatively included in the B. fungivorus-group of the genus Bursaphelenchus. Brief notes and morphometric data are given for the other species found. The record of B. mucronatus indicates that conditions for the establishment of the closely related B. xylophilus (the cause of pine wilt disease) may be suitable in Thailand and strict phytosanitary measures are therefore advisable.