The negative ramifications of invasive alien species (IAS) are considered the second-most cause of biodiversity extinction and endangerment after habitat modification. IAS movements are mainly anthropogenically driven (e.g., transport of shipping containers) and require fast detection to minimize damage and cost. The present study is the first to use molecular biosurveillance of international shipping containers to detect IAS and regulated species identification in Canada. Thirty-eight samples were collected from debris (soil, stems, seeds, individual specimens) found in containers arriving in Canada. A multi-marker approach using COI, ITS, ITS2, and 16S was used to identify four main taxonomic groups: arthropods, fungi, plants, and bacteria, respectively. Eleven IAS species were identified via metabarcoding based on environmental DNA samples, including two arthropods, six fungi, two plants, and one bacteria. The origin of the eDNA detected from each species was linked to their native distribution and country of origin, except for Lymantria dispar. Four physical specimens were also collected from shipping container debris and DNA barcoded, identifying three non-regulated species (two arthropods and one fungus). Altogether, these results demonstrate the importance of integrating molecular identification into current toolkits for the biosurveillance of invasive alien species and provide a set of validated protocols ready to be used in this context. Additionally, it reaffirms international shipping containers as a pathway for multiple invasive aliens and regulated species introduction in Canada. It also highlights the need to establish regular and effective molecular biosurveillance at the Canadian border to avoid new or recurrent invasions. Las ramificaciones negativas de las especies ex & oacute;ticas invasoras (EEI) se consideran la segunda causa de extinci & oacute;n y peligro de la biodiversidad despu & eacute;s de la modificaci & oacute;n del h & aacute;bitat. Los movimientos de EEI son impulsados principalmente por causas antropog & eacute;nicas (por ejemplo, transporte de contenedores de env & iacute;o) y requieren una detecci & oacute;n r & aacute;pida para minimizar da & ntilde;os y costos. El presente estudio es el primero en utilizar biovigilancia molecular de contenedores de env & iacute;o internacionales para detectar EEI y la identificaci & oacute;n de especies reguladas en Canad & aacute;. Se recolectaron treinta y ocho muestras de material (tierra, tallos, semillas, espec & iacute;menes individuales) encontrados en contenedores que llegaron a Canad & aacute;. Se utilizaron m & uacute;ltiples marcadores moleculares, COI, ITS, ITS2 y 16S, para identificar cuatro grupos taxon & oacute;micos principales: artr & oacute;podos, hongos, plantas y bacterias, respectivamente. Se identificaron once especies de EEI mediante matabarcoding basado en ADN ambiental, incluidos dos artr & oacute;podos, seis hongos, dos plantas y una bacteria. El origen del ADN ambiental detectado de cada especie estuvo vinculado a su distribuci & oacute;n nativa y pa & iacute;s de origen, excepto Lymantria dispar. Tambi & eacute;n se recolectaron cuatro espec & iacute;menes en los contenedores de env & iacute;o y se analizaron mediante c & oacute;digo de barras de ADN, identificando tres especies no reguladas (dos artr & oacute;podos y un hongo). En conjunto, estos resultados demuestran la importancia de integrar la identificaci & oacute;n molecular dentro de las herramientas actuales para la biovigilancia de especies ex & oacute;ticas invasoras y proporcionan un conjunto de protocolos validados listos para ser utilizados en este contexto. Adem & aacute;s, reafirma que los contenedores de transporte internacional son una v & iacute;a para la introducci & oacute;n de m & uacute;ltiples especies ex & oacute;ticas invasoras y especies reguladas en Canad & aacute;. Tambi & eacute;n destaca la necesidad de establecer una biovigilancia molecular peri & oacute;dica y eficaz en la frontera canadiense para evitar invasiones nuevas o recurrentes.

Winter temperatures are projected to increase in Central Europe. Subsequently, snow cover will decrease, leading to increased soil temperature variability, with potentially different consequences for soil frost depending on e.g. altitude. Here, we experimentally evaluated the effects of increased winter soil temperature variability on the root associated mycobiome of two plant species (Calluna vulgaris and Holcus lanatus) at two sites in Germany; a colder and wetter upland site with high snow accumulation and a warmer and drier lowland site, with low snow accumulation. Mesocosm monocultures were set-up in spring 2010 at both sites (with soil and plants originating from the lowland site). In the following winter, an experimental warming pulse treatment was initiated by overhead infrared heaters and warming wires at the soil surface for half of the mesocosms at both sites. At the lowland site, the warming treatment resulted in a reduced number of days with soil frost as well as increased the average daily temperature amplitude. Contrary, the treatment caused no changes in these parameters at the upland site, which was in general a much more frost affected site. Soil and plant roots were sampled before and after the following growing season (spring and autumn 2011). High-throughput sequencing was used for profiling of the root-associated fungal (ITS marker) community (mycobiome). Site was found to have a profound effect on the composition of the mycobiome, which at the upland site was dominated by fast growing saprotrophs (Mortierellomycota), and at the lowland site by plant species-specific symbionts (e.g. Rhizoscyphus ericae and Microdochium bolleyi for C. vulgaris and H. lanatus respectively). The transplantation to the colder upland site and the temperature treatment at the warmer lowland site had comparable consequences for the mycobiome, implying that winter climate change resulting in higher temperature variability has large consequences for mycobiome structures regardless of absolute temperature of a given site.