Assessment of the genetic diversity of the pinewood nematode, Bursaphelenchus xylophilus, in Portugal

O nemátode da madeira do pinheiro (NMP), Bursaphelenchus xylophiius, tem uma extensa distribuição na América do Norte, e encontra-se atualmente distribuído ao longo da maioria dos territórios de Canadá e dos Estados Unidos. Durante o último século, esta espécie foi transportada pelo Homem para outras regiões do mundo (não-nativas), associadas com o comércio e o fluxo global de produtos de origem florestal. Atualmente, esta espécie invasiva está reportada para algumas regiões do SE asiático (China, Japão, Coreia e Taiwan) e mais recentemente para a Europa (Portugal). Devido ao impacto que este organismo agente da doença da murchidão dos pinheiros causa nas florestas nativas destas regiões esta espécie assume uma elevada importância económica a nível mundial Em Portugal, a distribuição do NMP encontra-se confinada a uma área restrita e limitada (500 000 ha), a sul de Lisboa (península de Setúbal); contudo, constitui uma das maiores ameaças às florestas de pinheiro do país e da UE. Ate recentemente, nenhum consenso existia quanto à origem do NMP em Portugal. Diversas hipóteses têm sido colocadas para explicar esta introdução, nomeadamente a partir de zonas onde o nematode ocorre naturalmente (América do Norte), ou de outras áreas (não-nativas) onde o nematode se comporta como uma espécie invasiva (Leste da Ásia). A fim de avaliar a variabilidade genética do NMP proveniente da área afetada em Portugal, foram utilizadas várias técnicas moleculares, designadamente o random amplified polymorphic DNA (RAPD-PCR) e o satellite DNA (satDNA). No caso do RAPD-PCR, foram utilizados 24 isolados do NMP provenientes de Portugal, 1 proveniente da América do Norte e 1 da Ásia, tendo sido utilizado como out-group um isolado de B. mucronatus. A partir dos 28 RAPD primers utilizados obtiveram-se 640 fragmentos. No caso do satDNA, foram utilizados 21 isolados do NMP provenientes de Portugal, obtendo-se no total 206 sequências da família MspI. Ambos os métodos revelaram uma elevada similaridade genética entre os vários isolados do NMP da área afetada em Portugal O nível reduzido de diversidade genética obtido entre os isolados portugueses do NMP, permite concluir que se trata de uma única introdução deste organismo em Portugal, e proveniente de uma região asiática. A inexistência de uma de correlação entre a variabilidade genética e a distribuição geográfica do NMP dentro da área afetada em Portugal, indica que o NMP se encontra distribuído de forma uniforme ao longo de toda a área afetada, provavelmente relacionado com a distribuição e a expansão natural do inseto vector. The pinewood nematode (PWN), Bursaphelenchus xylophilus, has a wide distribution in North America, and is present throughout most of the territories of Canada and the United Stata. During the last century, this species has been transported by man to several non-native regions of the world, associated with trade and the global flow of forest products. Up to date, this invasive species has been reported from Asia (PR China, Japan, Korea and Taiwan) and more recently in Europe (Portugal). Due to the impact on native pine forests of these regions, this nematode species, the causal agent of pine wilt disease, is of great economic importance worldwide. In Portugal, the distribution of the PWN has been constrained to a relatively small area (500 000 ha) in the south of Lisbon (Setúbal Peninsula); however, it has become the most serious threat to pine forests in the country. Until recently, no consensus had emerged on the possible pathway of the PWN introduction in Portugal. Several hypotheses have been put forward to explain this introduction, such as an origin from endemic areas where the nematode naturally occurs (North America), or non-endemic areas where the nematode behaves as an exotic pest (East Asia). Random amplified polymorphic DNA (RAPD-PCR) and satellite DNA (satDNA) techniques were used in order to assess the level of genetic variability and genetic relationships, among several isolates of the PWN, representative of the entire affected area in Portugal. In the case of RAPD-PCR, 24 Portuguese isolates, plus two additional isolates of B. xylophilus, representing North America and East Asia were included. B. mucronatus was used as an out-group. Twenty-eight random primers generated a total of 640 DNA fragments. With satDNA, 206 Mspl sequence repeats were obtained from 21 Portuguese isolates of B. xylophilus. Both molecular methods revealed a high genetic similarity among the Portuguese isolates, and the low level of genetic diversity strongly suggests that they were dispersed recently from a single introduction, and from East Asia. The lack of apparent relationship between the genetic variability and the geographic distribution of the PWN within the affected area, suggests that the recent introduction of this pest (and pathogen) in Portugal has been uniformly distributed since its establishment, probably following the natural distribution and expansion of the insect vector.

Transferability of environmental favourability models in geographic space: The case of the Iberian desman (Galemys pyrenaicus) in Portugal and Spain

Transferring distribution models between different geographical areas may be problematic, as the performance of models outside their original scope is hard to predict. A modelling procedure is needed that gets the gist of the environmental descriptors of a distribution area, without either overfitting to the training data or overestimating the species’ distribution potential.We tested the transferability power of the favourability function, a generalized linear model, on the distribution of the Iberian desman (Galemys pyrenaicus) in the Iberian territories of Portugal and Spain.We also tested the effects of two of the main potential constraints on model transferability: the analysed ranges of the predictor variables, and the completeness of the species distribution data. We modelled 10 km×10km presence/absence data from Portugal and Spain separately, extrapolated each model to the other country, and compared predictions with observations. The Spanish model, despite arguably containing more false absences, showed good predictive ability in Portugal. The Portuguese model, whose predictors ranged between only a subset of the values observed in Spain, overestimated desman distribution when transferred.We discuss possible reasons for this differential model behaviour, and highlight the importance of this kind of models for prediction and conservation applications

Otter (Lutra lutra) distribution modeling at two resolution scales suited to conservation planning in the Iberian Peninsula

We used the results of the Spanish Otter Survey of 1994–1996, a Geographic Information System and stepwise multiple logistic regression to model otter presence/absence data in the continental Spanish UTM 10 10-km squares. Geographic situation, indicators of human activity such as highways and major urban centers, and environmental variables related with productivity, water availability, altitude, and environmental energy were included in a logistic model that correctly classified about 73% of otter presences and absences. We extrapolated the model to the adjacent territory of Portugal, and increased the model’s spatial resolution by extrapolating it to 1 1-km squares in the whole Iberian Peninsula. The model turned out to be rather flexible, predicting, for instance, the species to be very restricted to the courses of rivers in some areas, and more widespread in others. This allowed us to determine areas where otter populations may be more vulnerable to habitat changes or harmful human interventions. # 2003 Elsevier Ltd. All rights reserved.

Spatial, environmental and human influences on the distribution of otter (Lutra lutra) in the Spanish provinces

In a previous survey of otters ( Lutra lutra L. 1758) in Spain, different causes were invoked to explain the frequency of the species in each province. To find common causes of the distribution of the otter in Spain, we recorded a number of spatial, environmental and human variables in each Spanish province. We then performed a stepwise linear multiple regression of the proportion of positive sites of otter in the Spanish provinces separately on each of the three groups of variables. Geographic longitude, January air humidity, soil permeability and highway density were the variables selected. A linear regression of the proportion of otter presence on these variables explained 62.4% of the variance. We then used the selected variables in a partial regression analysis to specify which proportions of the variation are explained exclusively by spatial, environmental and human factors, and which proportions are attributable to interactions between these components. Pure environmental effects accounted for only 5.5% of the variation, while pure spatial and pure human effects explained 18% and 9.7%, respectively. Shared variation among the components totalled 29.2%, of which 10.9% was explained by the interaction between environmental and spatial factors. Human factors explained globally less variance than spatial and environmental ones, but the pure human influence was higher than the pure environmental one. We concluded that most of the variation in the proportion of occurrences of otter in Spanish provinces is spatially structured, and that environmental factors have more influence on otter presence than human ones; however, the human influence on otter distribution is less structured in space, and thus can be more disruptive. This effect of large infrastructures on wild populations must be taken into account when planning large-scale conservation policies

Spatial distribution patterns of the non-native European catfish, Silurus glanis, from multiple online sources - a case study for the River Tagus (Iberian Peninsula)

Effective management of invasive fishes depends on the availability of updated information about their distribution and spatial dispersion. Forensic analysis was performed using online and published data on the European catfish, Silurus glanis L., a recent invader in the Tagus catchment (Iberian Peninsula). Eighty records were obtained mainly from anglers’ fora and blogs, and more recently from www.youtube.com. Since the first record in 1998, S. glanis expanded its geographic range by 700 km of river network, occurring mainly in reservoirs and in high-order reaches. Human-mediated and natural dispersal events were identified, with the former occurring during the first years of invasion and involving movements of >50 km. Downstream dispersal directionality was predominant. The analysis of online data from anglers was found to provide useful information on the distribution and dispersal patterns of this non-native fish, and is potentially applicable as a preliminary, exploratory assessment tool for other non-native fishes.