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Due to climate change, many regions are experiencing progressively milder winters. Consequently, pest insects from warm regions, particularly those with some tolerance to low temperatures, could expand their geographic range into these traditionally colder regions. The palm borer moth (Paysandisia archon) is a Neotropical insect that in recent decades has reached Europe and Asia as one of the worst pests of palm trees. Little is known about its ability to tolerate moderately cold winters and, therefore, to colonize new areas. In this work, we characterized the cold tolerance of Paysandisia archon by measuring its thermal limits: median lethal-temperature, LT 50, chill-coma onset temperature, CT min , supercooling point, SCP, freezing time and freezing survival. We found that this species was able to survive short periods of complete freezing, with survival rates of 87% after a 30-min freezing exposure, and 33% for a 1 h-exposure. It is then a moderately freeze-tolerant species, in contrast to all other lepidopterans native to warm areas, which are freeze-intolerant. Additionally, we investigated whether this insect improved its cold tolerance after either short or long pre-exposure to sub-lethal low temperatures. To that end, we studied potential changes in the main thermo-tolerance parameters and, using X-ray Computed Tomography, also in the morphological components of pretreated animals. We found that short pre-exposures did not imply significant changes in the SCP and CT min values. In contrast, larvae with long pretreatments improved their survival to both freezing and low temperatures, and required longer times for complete freezing than the other groups. These long-term pre-exposed larvae also presented several morphological changes, including a reduction in water content that probably explained, at least in part, their longer freezing time and higher freezing survival. Our results represent the first cold tolerance characterization of this pest insect, which could be relevant to better design strategies to combat it., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

As traditionally cold areas become warmer due to climate change, temperature could no longer be a barrier to the establishment of non-native insects. This is particularly relevant for pest insects from warm and tropical areas, mainly those with some tolerance to moderately low temperatures, which could expand their range into these new locations. From this perspective, in this work we studied the morphological and biochemical responses of the Neotropical pest Paysandisia archon to low temperatures, as part of a possible strategy to colonize new areas. To that end, wild larvae were exposed for 7 days to either low (1 and 5 °C) or ambient (23 °C) temperatures. We then quantified the inner and outer morphological changes, by X-Ray Computer Tomography and Digital Holographic Microscopy, as well as the accumulation of metabolites acting as potential endogenous cryoprotectants, by Spectrophotometry. We found that Paysandisia archon developed a cold-induced response based on different aspects. On the one hand, morphological changes occurred with a significant reduction both in fluids susceptible to freezing and fat body, together with the thickening, hardening and increased roughness of the integument. On the other hand, we found an increase in the hemolymph concentration of cryoprotective substances such as glucose (6-fold) and glycerol (2-fold), while trehalose remained unchanged. Surprisingly, this species did not show any evidence of cold-induced response unless the environmental temperature was remarkably low (1 °C). These results could be useful to improve models predicting the possible spread of such a pest, which should incorporate parameters related to its resistance to low temperatures., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The red palm weevil (RPW), Rhynchophorus ferrugineus, is one of the worst palm pests worldwide. Our study aims to assess its internal and external morphological response to a sudden but transient decrease in the environmental temperature. Wild pre-pupae were subjected for 7 days to either low (5.0 ± 0.5 °C) or ambient temperature (23 ± 1 °C). Such conditions mimic a thermal anomaly happening in the larval stage most exposed to environmental factors. We quantified the changes undergone at: 1) the internal morphology, by X-Ray Computer Tomography (CT); 2) the 3-D integument' architecture, by Digital Holographic Microscopy (DHM); and 3) the glucose in hemolymph as a potential endogenous cryoprotectant. From X-ray CT we found that both pre-pupae subjected to cold and those remaining at ambient temperature follow a development where their fat body content decreases while a thick and dense cuticle is formed. There was no difference between both groups in the rate of change of fat body/dense tissues. Nevertheless, the cold group presents a slight developmental delay at the level of hemolymph content. Through DHM we again obtained that pre-pupae subjected to cold have not experienced a stop in their development. However, a more obvious developmental delay is now observed in this group at the level of the integumental roughness. Finally, regarding glucose, we found similar levels in control and ambient temperature larvae, while it was clearly increased in 51,7% of those subjected to cold. Our whole results provide morphological and biochemical evidence showing that the larval-pupal transition of the RPW continues almost undisturbed even during the quiescent state induced by a sudden and severe cold event. Nevertheless, a certain developmental delay is observed in both internal and external morphology. Additionally, the increased glucose level only found in the cold group suggests that glucose is part of the RPW cold tolerance strategy., (Copyright © 2020 Elsevier Ltd. All rights reserved.)