Characterization of Insecticide Response-Associated Transcripts in the Colorado Potato Beetle: Relevance of Selected Cytochrome P450s and Clothianidin.

The Colorado potato beetle (Leptinotarsa decemlineata (Say)) is known for its capacity to cause significant damages to potato crops worldwide. Multiple approaches have been considered to limit its spread including the use of a diverse arsenal of insecticides. Unfortunately, this insect frequently develops resistance towards these compounds. Investigating the molecular bases underlying the response of L. decemlineata against insecticides is of strong interest to ultimately devise novel and targeted approaches aimed at this pest. This work aimed to characterize, via qRT-PCR, the expression status of targets with relevance to insecticide response, including ones coding for cytochrome P450s, glutathione s-transferases, and cuticular proteins, in L. decemlineata exposed to four insecticides; chlorantraniliprole, clothianidin, imidacloprid, and spinosad. Modulation of levels associated with transcripts coding for selected cytochrome P450s was reported in insects treated with three of the four insecticides studied. Clothianidin treatment yielded the most variations in transcript levels, leading to significant changes in transcripts coding for CYP4c1, CYP4g15, CYP6a13, CYP9e2, GST, and GST-1-Like. Injection of dsRNA targeting CYP4c1 and CYP9e2 was associated with a substantial decrease in expression levels and was, in the case of the latter target, linked to a greater susceptibility of L. decemlineata towards this neonicotinoid, supporting a potential role for this target in clothianidin response. Overall, this data further highlights the differential expression of transcripts with potential relevance in insecticide response, as well as generating specific targets that warrant investigation as novel dsRNA-based approaches are developed against this insect pest.

Exposure to Temperature and Insecticides Modulates the Expression of Small Noncoding RNA-Associated Transcripts in the Colorado Potato Beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae).

The Colorado potato beetle (Leptinotarsa decemlineata (Say)) is an insect that can adapt to various challenges, including temperature fluctuations or select insecticide treatments. This pest is also an ongoing threat to the potato industry. Small noncoding RNAs such as miRNAs, which can control posttranscriptionally the expression of various genes, and piRNAs, which can notably impact mRNA turnover, are modulated in insects under different conditions. Unfortunately, information regarding the expression status of key players involved in their synthesis and function is for the most part lacking. The current study thus aims at assessing the levels of such targets in L. decemlineata exposed to hot and cold temperatures as well as treated to the insecticides chlorantraniliprole, clothianidin, imidacloprid, and spinosad. Transcript expression levels of Ago1, Ago2, Ago3, Dcr2a, Dcr2b, Expo-5, Siwi-1, and Siwi-2, components of pathways associated with small noncoding RNA production or function, were measured by qRT-PCR and revealed modulation of select transcripts in response to temperature challenges and to select insecticides. RNAi-mediated reduction of Ago2 transcript levels in L. decemlineata injected with Ago2-targeting dsRNA and exposed to cold and warm temperatures was also conducted. Changes in survival rates were observed for the latter condition in dsRNA- versus saline-injected insects. These results showcase the differential expression of select targets involved in small noncoding RNA homeostasis and provide leads for the subsequent assessment of their involvement during stress response in L. decemlineata using RNAi-based approaches.

Transcriptomics-Based Approach Identifies Spinosad-Associated Targets in the Colorado Potato Beetle, Leptinotarsa decemlineata.

The Colorado potato beetle Leptinotarsa decemlineata is an insect pest that threatens potato crops globally. The primary method to control its damage on potato plants is the use of insecticides, including imidacloprid, chlorantraniliprole and spinosad. However, insecticide resistance has been frequently observed in Colorado potato beetles. The molecular targets and the basis of resistance to imidacloprid and chlorantraniliprole have both been previously quantified. This work was undertaken with the overarching goal of better characterizing the molecular changes associated with spinosad exposure in this insect pest. Next-generation sequencing was conducted to identify transcripts that were differentially expressed between Colorado potato beetles exposed to spinosad versus control insects. Results showed several transcripts that exhibit different expression levels between the two conditions, including ones coding for venom carboxylesterase-6, chitinase 10, juvenile hormone esterase and multidrug resistance-associated protein 4. In addition, several microRNAs, such as miR-12-3p and miR-750-3p, were also modulated in the investigated conditions. Overall, this work reveals a molecular footprint underlying spinosad response in Colorado potato beetles and provides novel leads that could be targeted as part of RNAi-based approaches to control this insect pest.

Temperature affects the reproductive outputs of coral-eating starfish Acanthaster spp. after adult exposure to near-future ocean warming and acidification.

Outbreaks of the coral-eating crown-of-thorns starfish Acanthaster spp. (COTS) have become to be amongst the most severe threats to coral reefs worldwide. Although most research has focused on COTS early development, it remains unclear how COTS populations will keep pace with changing ocean conditions. Since reproduction is a key process contributing to outbreaks, we investigated the reproductive success of adult COTS acclimated for 3-4 months to different treatment combinations of ambient conditions, ocean warming (+2 °C) and acidification (-0.35 pH). Our results suggest that the optimal breeding season in New Caledonia is concentrated around the end of the calendar year, when water temperature reaches >26 °C. We found negative effects of temperature on egg metrics, fertilisation success, and GSI, conflicting with previously documented effects of temperature on echinoderm reproductive outputs. Fertilisation success dropped drastically (more than threefold) with elevated temperature during the late breeding season. In contrast, we detected no effects of near-future acidification conditions on fertilisation success nor GSI. This is the first time that COTS reproduction is compared among individuals acclimated to different conditions of warming and acidification. Our results highlight the importance of accounting for adult exposure to better understand how COTS reproduction may be impacted in the face of global change.

Citizen Science, a promising tool for detecting and monitoring outbreaks of the crown-of-thorns starfish Acanthaster spp.

Monitoring potentially devastating coral-eating crown-of-thorns starfish (COTS) populations at scales relevant to management is a challenging task. Here, we investigated a citizen science approach to detect COTS outbreaks and prioritize management responses. Between 2014 and 2018, 38 000 COTS were recorded through 641 online observation reports submitted across New Caledonia, Vanuatu and Fiji by private stakeholders (51%), NGOs (22%), business operator (11%), research/government agencies (16%). COTS were observed in multiple areas, including in remote/inhabited reefs where they had never been reported. A three-level classification was developed to discriminate risk areas and propose operational guidelines to streamline management actions. About two-thirds of reports had low abundances (<10 starfish sighted) and could be addressed with low priority. Verification surveys at 65 reef sites confirmed outbreaks in half of the cases, along with high peak densities (7 000 ind.ha-1). Combining professional and non-professional observers increased the detection range (+27%) and the number of COTS detections (+129%). Citizen reports were eventually followed by removal campaigns organized within diverse institutional frameworks. While citizen monitoring has intrinsic limitations, we advocate that it constitutes a complementary and promising approach to support the ongoing management efforts in all countries affected by COTS.

Differential expression of transcripts with potential relevance to chlorantraniliprole response in the Colorado potato beetle, Leptinotarsa decemlineata.

The Colorado potato beetle (Leptinotarsa decemlineata [Say]) is an insect pest that can significantly harm potato plants worldwide. Control of this insect relies heavily on chemical insecticides such as chlorantraniliprole. Nevertheless, the complete molecular signature associated with response to this compound is lacking in L. decemlineata. In this study, amplification and quantification by qRT-PCR (quantitative reverse transcription-polymerase chain reaction) of targets relevant to chlorantraniliprole were undertaken in insects exposed to this chemical. This approach showed modulation of numerous cytochrome P450s, such as CYP350D1 and CYP4Q3, as well as upregulation of microRNAs (miRNAs), including miR-1-3p and miR-305-5p, in chlorantraniliprole-exposed insects. Functional assessment of transcript targets predicted to be regulated by these miRNAs was performed and revealed their likely impact on transcriptional regulation. RNAi-based targeting of CYP350D1 notably provided preliminary evidence of its underlying implication for chlorantraniliprole response in L. decemlineata. Overall, this study strengthens the current knowledge of the molecular changes linked to chlorantraniliprole response in L. decemlineata and provides novel targets with potential relevance to chlorantraniliprole susceptibility in this insect pest of global relevance.

Expression status of heat shock proteins in response to cold, heat, or insecticide exposure in the Colorado potato beetle Leptinotarsa decemlineata.

The Colorado potato beetle (Leptinotarsa decemlineata (Say)) is an agricultural pest that threatens the potato industry worldwide. This insect is widely regarded as one of the most difficult-to-control pests, as it can thrive in a wide range of temperature conditions and routinely develops resistance towards various insecticides. The molecular changes associated with response to these challenges have not been fully investigated in L. decemlineata. While differential expression and characterization of heat shock proteins (HSPs) in response to stress have been conducted in several insects, data regarding HSPs in L. decemlineata are limited. The overarching objective of this study consisted of evaluating the expression of various HSPs in L. decemlineata exposed to different temperatures or treated with the insecticides imidacloprid and chlorantraniliprole. Expression levels of HSP60, HSP70, HSP90, and HSP Beta-1 were evaluated by qRT-PCR and insect mortality was assessed using dsRNAs aimed at select HSP targets. Elevated HSP70 and HSP90 transcript levels were observed in heat-exposed L. decemlineata while downregulation of HSP70 transcript levels was measured in insects submitted to cold conditions. Chlorantraniliprole exposure was associated with reduced HSP Beta-1 transcript levels while no change in expression was monitored in insects exposed to imidacloprid. RNAi-based knockdown of HSP60 levels correlated with significant insect mortality 14 days after dsRNA injection. These results highlight the modulation of HSPs that occur in L. decemlineata exposed to fluctuating temperatures and position HSPs as interesting candidates in the identification of novel molecular leads that could be targeted to control this insect.

Ephemeral and Localized Outbreaks of the Coral Predator Acanthaster cf. solaris in the Southwestern Lagoon of New Caledonia.

Mehdi Adjeroud, Mohsen Kayal, Christophe Peignon, Matthieu Juncker, Suzanne C. Mills, Ricardo Beldade, and Pascal Dumas (2018) Outbreaks of the coral predator Acanthaster spp., the crown-of-thorns seastar (COTS), cause major coral declines across the Indo-Pacific. However, the processes surrounding the initiation and propagation of COTS outbreaks are still unclear. We observed COTS outbreak abundances on several mid-shelf and inner-barrier reefs in the southern section of the New Caledonian lagoon, a multi-location initiation event that we expected to precede a broader region-wide disturbance. However, reef monitoring over 3 years revealed the highly localized and ephemeral character of these outbreaks. Outbreaks that were observed at four reef locations at the beginning of the survey simply faded away, without any specific management actions such as culling efforts. We also found no distinct reef biotope on which COTS outbreaks originated, although mid-shelf and inner-shelf barrier reefs seem to be favoured. New Caledonia has been exempted from to widespread regional COTS outbreaks, which is surprising given its proximity to the Australian Great Barrier Reef - a reef complex particularly vulnerable to COTS - and Vanuatu - where large-scale outbreaks were recorded during the same period. The availability of coral prey is probably not a limiting factor for the propagation of COTS outbreaks on New Caledonian reefs, given the high abundance and diversity of coral assemblages. Our findings reveal that localized and ephemeral COTS outbreaks can be naturally contained and do not necessarily result in widespread disturbances.

Tectus (Trochus) niloticus search for suitable habitats can cause equivocal benefits of protection in village-based marine reserves.

In the Pacific, the protection of coral reef resources is often achieved through the implementation of village-based marine reserves (VBMRs). While substantial fisheries benefits are often reported, results of quantitative approaches are controversial for benthic macroinvertebrates, whose life history traits may cause low congruence with protective measures implemented at non-ecologically relevant scales. This study investigated the structural and behavioral responses of the exploited topshell Tectus niloticus within a very small (0.2 km2) VBMR in Vanuatu, south Pacific. The results of underwater surveys and a nine-month tagging experiment emphasized contrasted, scale-dependent responses. At the reserve scale, our results failed to demonstrate any positive effect of protection after three years of closure. In contrast, abundance, density and biomass increased more than ten-fold in the southern part of the reserve, along with significantly larger (25%) individual sizes. The dispersal of tagged specimens was also consistently lower after 2, 4 and 9 months in the latter zone. Analyses of 17 substratum variables revealed a marked small-scale patchiness delineating contrasted benthic microhabitats, the distribution of which closely matched that of trochus. We advocate that i) VBMRs have inherently unequal ecological potentials for protecting and managing highly habitat-dependent species such as trochus; ii) 'success' or 'failure' is to a certain extent pre-determined by the trajectory of species-specific microhabitats, which may outreach protection effects. This has strong implications in the Pacific where the location and size of reserves primarily depends upon marine tenure, and communities have little flexibility in setting reserve boundaries.

Circadian behaviour of Tectus (Trochus) niloticus in the southwest Pacific inferred from accelerometry.

BACKGROUND: Behaviour and time spent active and inactive are key factors in animal ecology, with important consequences for bioenergetics. For the first time, here, we equipped the gastropod Tectus (= Trochus) niloticus with accelerometers to describe activity rhythms at two sites in the Southwest Pacific with different temperature regimes: New Caledonia and Vanuatu. RESULTS: Based on a 24-hour cycle, T. niloticus activity began at dusk and gradually stopped during the night, before sunrise. This nocturnal behaviour was characterised by short (duration <30 s), low intensity (acceleration < 0.12 É¡) movements and was probably associated with foraging behaviour. We assumed that activity ceased once the animal was satiated. Our analysis of two size groups in Vanuatu (80-90 mm vs. 120-140 mm, basal shell diameter) revealed a size effect; smaller specimens displayed greater activity, reflected by more intense and longer movements while migrating at night toward the edge of the reef. This nocturnal behaviour is not uncommon for grazing gastropods and is mainly associated with attempting to avoid visual predators whilst feeding. CONCLUSIONS: The use of accelerometers coupled with light and temperature sensors provided detailed information on topshell behaviour and physiology under natural conditions. These data provide a foundation for identifying potential changes in the fine-scale behaviour of T. niloticus in response to environmental changes, which is essential in animal ecology and stock conservation.

Lime Juice and Vinegar Injections as a Cheap and Natural Alternative to Control COTS Outbreaks.

Outbreaks of the corallivorous crown-of-thorns seastar Acanthaster planci (COTS) represent one of the greatest disturbances to coral reef ecosystems in the Indo-Pacific, affecting not only coral reefs but also the coastal communities which rely on their resources. While injection approaches are increasingly used in an attempt to control COTS densities, most of them display severe drawbacks including logistical challenges, high residual environmental impacts or low cost-effectiveness. We tested a new alternative control method based upon acidic injections of cheap, 100% natural products. We investigated the lethal doses, intra- and inter-specific disease transmission and immune responses of COTS when injected with fresh lime juice (extracted from local Citrus arantifolia) and white spirit vinegar. High COTS mortality was achieved with small volumes: 10-20 ml per seastar induced death in 89%/97% of injected specimens after an average 34.3 h/29.8 h for lime juice and vinegar respectively. Highest efficiency was reached for both solutions with double shots of (2 × 10 ml) in two different areas on the body: 100% mortality occurred within 12-24 h, which is similar or faster compared with other current injection methods. Multiple immune measures suggested that death was very likely caused by pH stress from the acidic solutions rather than a bacterial infection. Contagion to either conspecifics or a variety of other reef species was not observed, even at COTS densities 15 times higher than the highest naturally reported. 10 to 20 l lime juice/vinegar could kill up to a thousand COTS at a cost of less than 0.05 USD per specimen; no permits or special handling procedures are required. We conclude that injections of lime juice and vinegar offer great advantages when compared to current best practises and constitute a cheap and natural option for all reefs affected by COTS.

Wildfire risk for main vegetation units in a biodiversity hotspot: modeling approach in New Caledonia, South Pacific.

Wildfire has been recognized as one of the most ubiquitous disturbance agents to impact on natural environments. In this study, our main objective was to propose a modeling approach to investigate the potential impact of wildfire on biodiversity. The method is illustrated with an application example in New Caledonia where conservation and sustainable biodiversity management represent an important challenge. Firstly, a biodiversity loss index, including the diversity and the vulnerability indexes, was calculated for every vegetation unit in New Caledonia and mapped according to its distribution over the New Caledonian mainland. Then, based on spatially explicit fire behavior simulations (using the FLAMMAP software) and fire ignition probabilities, two original fire risk assessment approaches were proposed: a one-off event model and a multi-event burn probability model. The spatial distribution of fire risk across New Caledonia was similar for both indices with very small localized spots having high risk. The patterns relating to highest risk are all located around the remaining sclerophyll forest fragments and are representing 0.012% of the mainland surface. A small part of maquis and areas adjacent to dense humid forest on ultramafic substrates should also be monitored. Vegetation interfaces between secondary and primary units displayed high risk and should represent priority zones for fire effects mitigation. Low fire ignition probability in anthropogenic-free areas decreases drastically the risk. A one-off event associated risk allowed localizing of the most likely ignition areas with potential for extensive damage. Emergency actions could aim limiting specific fire spread known to have high impact or consist of on targeting high risk areas to limit one-off fire ignitions. Spatially explicit information on burning probability is necessary for setting strategic fire and fuel management planning. Both risk indices provide clues to preserve New Caledonia hot spot of biodiversity facing wildfires.

Small-scale habitat structure modulates the effects of no-take marine reserves for coral reef macroinvertebrates.

No-take marine reserves are one of the oldest and most versatile tools used across the Pacific for the conservation of reef resources, in particular for invertebrates traditionally targeted by local fishers. Assessing their actual efficiency is still a challenge in complex ecosystems such as coral reefs, where reserve effects are likely to be obscured by high levels of environmental variability. The goal of this study was to investigate the potential interference of small-scale habitat structure on the efficiency of reserves. The spatial distribution of widely harvested macroinvertebrates was surveyed in a large set of protected vs. unprotected stations from eleven reefs located in New Caledonia. Abundance, density and individual size data were collected along random, small-scale (20×1 m) transects. Fine habitat typology was derived with a quantitative photographic method using 17 local habitat variables. Marine reserves substantially augmented the local density, size structure and biomass of the target species. Density of Trochus niloticus and Tridacna maxima doubled globally inside the reserve network; average size was greater by 10 to 20% for T. niloticus. We demonstrated that the apparent success of protection could be obscured by marked variations in population structure occurring over short distances, resulting from small-scale heterogeneity in the reef habitat. The efficiency of reserves appeared to be modulated by the availability of suitable habitats at the decimetric scale ("microhabitats") for the considered sessile/low-mobile macroinvertebrate species. Incorporating microhabitat distribution could significantly enhance the efficiency of habitat surrogacy, a valuable approach in the case of conservation targets focusing on endangered or emblematic macroinvertebrate or relatively sedentary fish species.

Coral assemblages in Tonga: spatial patterns, replenishment capacities, and implications for conservation strategies.

Coral reefs in Tonga have been confronted by multiple threats of various origins, including large-scale disturbances and human-induced stressors. These reef communities have been poorly studied, and efficient conservation actions are urgently needed. The aim of this study was to: (1) examine the spatial distribution of coral assemblages in the lagoon of Tongatapu; (2) determine the degree to which spatial heterogeneity of adult corals is influenced by recruitment processes; and (3) examine the implications of these results in terms of conservation actions. We recorded a total of 37 adult and 28 juvenile coral genera, a mean density of 11.6 adult and 5.5 juvenile colonies m(-2), and a dominance of Montipora, Acropora, and Porites. For seven of the 10 dominant genera, spatial patterns of adults were linked to the short-term recruitment pattern history. Despite a reduced diversity and abundance of adult corals in some areas, the lagoon of Tongatapu retains the potential for replenishment through recruitment of young corals. Consequently, we suggest that conservation actions should focus on reducing factors causing coral mortality and maintain suitable conditions for the establishment and growth of juvenile corals, thus increasing the probability that they will reach maturity and participate to the maintenance of local populations. Rather than establishing a large marine protected area, which will almost certainly suffer from a lack of control and poor enforcement, alternative conservation measures could be successfully implemented through the establishment of several small village-based marine reserves, as has been undertaken in other South Pacific islands with promising results.

Developing erosion models for integrated coastal zone management: a case study of The New Caledonia west coast.

The tropical climate and human pressures (mining industry, forest fires) cause significant sediment inputs into the New Caledonia lagoon and are a major cause of degradation of the fringing reefs. The erosion process is spatially characterized on the west coast of New Caledonia to assess potential sediment inputs in the marine area. This paper describes the methodologies that are used to map soil sensitivity to erosion using remote sensing and a geographic information system tool. A cognitive approach, multi-criteria evaluation model and Universal Soil Loss Equation are implemented. This article compares the relevance of each model in order to spatialize and quantify potential erosion at catchment basin scale. These types of studies provide valuable results for focusing on areas subject to erosion and serve as a decision-making tool for the minimization of lagoon vulnerability to the natural and human dynamics on the level of the catchment basins.