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.

Context Invasive plants are one of the most significant threats to woodlands globally. Methods of invasive plant control include manual removal and herbicide application. While the impacts of control methods on invasive and off-target native plant species are often explored, the impacts on below-ground organisms, such as fungi, are less well understood.Aims We conducted a glasshouse trial to investigate the responses of soil fungal communities to herbicides and manual removal that are used to control common invasive plant species in Banksia woodland in south-western Australia.Methods Broad spectrum (glyphosate and pelargonic acid) and grass-specific (fluazifop-p-butyl) herbicides were separately applied to pots containing either Ehrharta calycina, a key invasive grass species or Eucalyptus todtiana, a native woodland tree at the recommended woodland rate. After six weeks, samples of treated soils were subjected to high throughput sequencing to determine fungal community diversity, richness, relative abundance, composition and putative ecosystem function.Key results Pelargonic acid induced the widest range of changes including decreased fungal richness and Shannon diversity but all herbicides affected community composition. Within functional groups, fluazifop-p-butyl led to a significant decrease of symbiotrophs in the mycorrhizal species.Conclusions We show that invasive species management, in the manner applied, can lead to immediate changes in fungal community composition.Implications Observed patterns require further exploration, particularly repeat testing under different environmental conditions, to better determine the impact and mode of action of herbicides on below-ground organisms. The functional changes in the soil fungal community could further disturb the soil fungal community and complicate subsequent management considerations.

Vascular wilt is an important tomato disease that affects culture yields worldwide, with Fusarium oxysporum (F.o) being the causal agent of this infection. Several management strategies have lost effectiveness due to the ability of this pathogen to persist in soil and its progress in vascular tissues. However, nowadays, research has focused on understanding the plant defense mechanisms to cope with plant diseases. One recent and promising approach is the use of extracellular DNA (eDNA) based on the ability of plants to detect their self-eDNA as damage-associated molecular patterns (DAMPs) and pathogens' (non-self) eDNA as pathogen-associated molecular patterns (PAMPs). The aim of this work was to evaluate the effect of the eDNA of F.o (as a DAMP for the fungus and a PAMP for tomato plants) applied on soil, and of tomato's eDNA (as a DAMP of tomato plants) sprayed onto tomato plants, to cope with the disease. Our results suggested that applications of the eDNA of F.o (500 ng/mu L) as a DAMP for this pathogen in soil offered an alternative for the management of the disease, displaying significantly lower disease severity levels in tomato, increasing the content of some phenylpropanoids, and positively regulating the expression of some defense genes. Thus, the eDNA of F.o applied in soil was shown to be an interesting strategy to be further evaluated as a new element within the integrated management of vascular wilt in tomato.

Environmental DNA (eDNA) carrying antibiotic resistance gene (ARG) has attracted a great deal of attention because of its threat to the ecology and human health. Traditional porous adsorbents, such as microporous biochar and natural mineral, are low-effective in removing eDNA from sewage. This study used cuttlefish-bone (CB), a fishery waste, as an anticipated material to adsorb a model compound of eDNA from herring sperm (hsDNA). An interesting result was firstly observed that extremely high DNA adsorption on cuttlefish-bone pyrolysis derivative (CCB) was up to 88.7 mg/g, 3-10 folds higher than that of most other adsorbents in the existing literatures, which was attributed to the carbon film and large pores. To achieve an adsorption rate of 75 %, hsDNA adsorption took 96 h on CB but only 24 h on CCB, which was attributed to the fluent channel of CCB. The ligand exchange, Ca2+ bridge and pi-pi interaction were identified as dominated adsorption mechanisms, based on FTIR and phosphate competition experiments. This study exploited a high-efficient, environmentally friendly, and low-cost adsorbent for treating ARG-contaminated soil and water.