Plant bio-protection and biosecurity programs worldwide use trap-based surveillance for the early detection of agricultural pests and pathogens to contain their incursions and spread. This task is reliant on effective preservation in insect traps, which is required to maintain specimen quality for extended periods under variable environmental conditions. Furthermore, with traditional morphological examinations now increasingly paired with modern molecular diagnostic techniques, insect traps are required to preserve both the specimens’ morphology and the DNA of insects and vectored bacterial pathogens. Here, we used psyllids (Hemiptera) and their hosted bacteria as a model to test the preservative ability of propylene glycol (PG): a non-flammable, easily transportable preservative agent that could be used in pitfall, suction or malaise traps. We tested preservation using various PG concentrations, at different temperatures and for multiple time periods, paired with non-destructive DNA extraction methods, which allow isolation of both insect and arbobacterial DNA while retaining a morphological voucher of the insect host specimens. PG concentrations between 40% and 100% performed best for both insect and bacterial DNA preservation up to 28 days. Ultimately, given the viscous nature of PG at high concentrations, we recommend using it at a concentration between 40% and 60% to enable insects to sink into the solution, thus enhancing DNA preservation.

Insects preserved as reference specimens are important in a wide range of fields, including health, pest management and forensics. The aim of the present study was to test a non‐destructive DNA extraction method on samples of soft‐bodied insects, fly larvae, which are otherwise hard to identify morphologically. This not only provides DNA enabling molecular identification but also retains morphological reference specimens for samples belonging to collections and museums that cannot be destroyed. In this work, fly larvae identified as belonging to the family Muscidae were non‐destructively processed. DNA barcoding molecular identification allowed most of these specimens to be assigned to species. Furthermore, the visibility of seven important larval morphological characters – anterior and posterior spiracles, mouth hooks, cephalopharyngeal skeleton, locomotory welts, segmentation and colour – was assessed pre‐ and post‐DNA extraction. It was found that the morphology generally did not deteriorate post‐DNA extraction but actually improved through increased visibility of internal features. Therefore, this non‐destructive DNA extraction method not only allowed COI barcode sequences to be obtained, but also enabled a better morphological identification of the fly larvae retaining physical reference voucher specimens and avoiding the need for dissections.

Pathogens which need a vector for their transmission can alter the vectors’ behaviour to favour their spread. We used the electrical penetration graph technique to investigate this hypothesis by using the tomato potato psyllidBactericera cockerelliinfected or not with the plant pathogenCandidatusLiberibacter solanacearum (CLso) on African boxthorn and tomato. Probing was not affected by the host type but there was a significant effect on probing due to the infection status of the psyllid. More psyllids carried out probing activities in the sieve elements when infected with CLso, and more probing activities were observed from CLso-infected psyllids by comparison to the non-infected groups. Specifically, significant increases in salivation, phloem ingestion and number of probes, before and after reaching the sieve elements, were noticed in the infected groups. Furthermore, time elapsed to reach the sieve elements was significantly shortened by 2 h in the infected group. Remaining probing activities in xylem tissues were not different between all psyllid groups. The observed changes in feeding behaviour by pathogen-infected psyllids may well ensure further spread of the pathogen as greater salivation has the potential to increase transmission, highlighting at the same time the important role that crop and non-crop hosts play in disease epidemiology.

Fruit flies are considered economically important insects due to some species being agricultural pests. However, morphological identification of fruit fly adults and larvae can be difficult requiring a high level of taxonomic expertise, with misidentifications causing problematic false‐positive/negative results. While destructive molecular techniques can assist with the identification process, these often cannot be applied where it is mandatory to retain a voucher reference specimen. In this work, we non‐destructively (and partial‐destructively) processed larvae and adults mostly belonging to the species Dirioxa pornia (Walker, 1849), of the poorly studied nonpest fruit fly tribe Acanthonevrini (Tephritidae) from Australia, to enable molecular identifications whilst retaining morphological vouchers. By retaining the morphological features of specimens, we confirmed useful characters for genus/species‐level identification, contributing to improved accuracy for future diagnostics using both molecular and morphological approaches. We provide DNA barcode information for three species of Acanthonevrini known from Australia, which prior to our study was only available for a single species, D. pornia. Our specimen examinations provide new distribution records for three nonpest species: Acanthonevroides variegatus Permkam and Hancock, 1995 in South Australia, Acanthonevroides basalis (Walker, 1853) and D. pornia in Victoria, Australia; as well as new host plant records for D. pornia, from kangaroo apple, apricot and loquat.

The bird cherry-oat aphid, Rhopalosiphum padi (L.), is an economically significant pest of pasture grasses, the latter being capable of hosting several fungal endophyte–perennial ryegrass symbiota rich in alkaloids and toxic to vertebrates and invertebrates. Measuring aphid feeding behaviour can provide insights into the effectiveness and mode of action of different fungal endophytes. This study investigated the effects of different Epichloë–perennial ryegrass symbiota on the feeding behaviour of R. padi using the electrical penetration graph technique while also assessing the aphid life history. In most cases, endophytes had significant feeding deterrence and paired fecundity and mortality effects. But, in some instances, endophytes with the highest aphid mortality did not significantly deter feeding, suggesting a more complicated scenario of interactions between the relative concentration of metabolites, e.g., host plant defence response metabolites and alkaloids, and/or physical changes to leaf morphology. Overall, this study sheds light on the mode of action of Epichloë endophytes against aphids and highlights the importance of Epichloë–perennial ryegrass symbiota in the management of insect pests such as aphids in pasture-based grazing systems.