Rapid and portable molecular detection tools for reptile pest priority species in Australian environmental biosecurity scenarios

The global transnational trade in exotic pets has resulted in the introduction of many invasive alien species into novel environments. Reptiles are considered particularly high risk, with many global examples of their detrimental impacts on both local environments and economies. The trade in exotic pets facilitates the transport of non-native species, which can lead to the establishment of invasive alien species. Introductions and subsequent establishment can be classified as either unintentional (i.e., stowaway or hitchhiker) or intentional (through unwanted release). In Australia, the trade in exotic pets has resulted in the introduction of a range of species, including two high-risk terrestrial reptiles: common boa (Boa constrictor), and the American corn snake (Pantherophis guttatus). Predicted consequences of their introduction, establishment and spread are dire for Australian biodiversity outcomes due to their broad climate tolerance, generalist ecology, and previous history of establishment success. Consequently, both species have been identified for environmental biosecurity prioritisation and included as highest risk on the Australian Government Department of Agriculture, Fisheries, and Forestry National Priority List of Exotic Environmental Pests, Weeds and Diseases. Limited attention has been directed at compliance tools for environmental biosecurity monitoring of post-border trade. I identified Loop-Mediated Isothermal Amplification (LAMP) as an emerging tool for the DNA detection of high-risk species in environmental biosecurity and compliance contexts. To help combat the trade of B. constrictor and P. guttatus I have developed fit for purpose LAMP assays for environmental biosecurity compliance. I rigorously assessed key practical and analytical factors, including: (i) appropriate sample types; (ii) the recovery and detection of trace DNA from reptile contact; (iii) suitable DNA extraction methods; (iv) detection parameters and chemistries; (v) sensitivity/specificity; and (vi) usability of portable detection tools, applying LAMP to DNA detection of high-risk pest reptiles. My findings strengthen the case for isothermal amplification methods as rapid presumptive testing tools applied to environmental biosecurity. The described LAMP assays facilitate detection of key incursion reptiles in less than 30 mins using in-laboratory and portable technologies. With continued development and integration into the biosecurity toolkit, these compliance tools have a great capacity for presumptive portable detection.

Author dc:creator

• Deliveyne, Nathan

Advisors dc:contributor.advisor

• Cassey, Phillip
• Austin, Jeremy J.
• Sanders, Kate L.

• Environmental biosecurity
• invasive species
• Loop-Mediated Isothermal Amplification (LAMP)
• wildlife forensics
• reptiles

Language dc:language.iso

en

Handle dc:identifier.uri

https://hdl.handle.net/2440/144549

OAI identifier oai:identifier

oai:digital.library.adelaide.edu.au:2440/144549