As a result of the detection of the pinewood nematode Bursaphelenchus xylophilus in Portugal, and its subsequent spread to Spain, intense surveys were conducted to screen for the presence of Bursaphelenchus species in Romania. Herein, we report recent surveys of insects potentially vectoring Bursaphelenchus species collected using trap trees or pheromone‐baited traps placed in the forest. Trap felled spruce trees (Picea abies) and pheromone‐baited traps were installed in six different counties in Romania (Brașov, Sibiu, Suceava, Hunedoara, Timiș and Dâmbovița). Ten different species of insects distributed among Curculionidae and Cerambycidae were obtained. Nematodes were extracted from insects and observed to validate the presence of Bursaphelenchus specimens. One female identified as Monochamus sutor was the only specimen carrying nematodes in the genus Bursaphelenchus. Nematodes were identified as B. mucronatus based on morphological and molecular features. This is the first detection and report of natural spread of B. mucronatus in Romania. The absence of B. xylophilus was confirmed in the areas of Romania surveyed in this work.
Six species of the genus Bursaphelenchus were recovered as the result of a survey to assess the potential introduction of Bursaphelenchus xylophilus into the conifer forests of Romania. Bursaphelenchus xylophilus was not detected in the sampling regions. Bursaphelenchus dietrichi was recovered from one point, representing the first report of this species in the country. The other detected species, B. abietinus, B. andrassyi, B. hofmanni, B. fraudulentus and B. willibaldi, were characterized by their morphometric data and molecular analyses. The morphometrics of Romanian populations of B. hofmanni and B. willibaldi are presented for the first time. The phylogenetic relationships of the recovered species (except for B. abietinus, with no successful sequencing) were reconstructed using the sequences of the D2‐D3 expansion segments of the large subunit of ribosomal DNA (D2‐D3LSU rDNA). Some of the species were also characterized using the restriction fragment length polymorphism (RFLP) pattern of the internal transcribed spacer (ITS) fragment. The distribution, related host trees and phylogenetic relationships of the identified species are discussed.
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