Communities of metazoan parasites in seven sympatric skate species (Elasmobranchii, Rajidae) from the English Channel and Celtic Sea differing in conservation status.

Elasmobranch populations are in steep decline mainly due to overfishing bycatch, but parasites may accelerate the collapse of vulnerable and/or highly parasitized species. We therefore studied metazoan parasites of Rajidae from the northeast Atlantic: vulnerable Leucoraja fullonica, near-threatened Raja brachyura, Raja clavata, Raja microocellata and Raja undulata, and least-concerned Raja montagui and Leucoraja naevus. Overall prevalence varied from 19% for R. montagui to 100% for L. fullonica. Parasite communities differed between skate species, and prevalence and abundance were higher for L. fullonica, R. microocellata, and R. undulata. We recorded 11 parasite taxa in the study: three nematodes, six cestodes, one monogenean, and one myxosporean. Whatever the skate species, the parasite component community comprised at least two nematode taxa among Phocanema spp., Proleptus sp. and Anisakis simplex. DNA-sequencing revealed that Phocanema azarasi and Phocanema krabbei both occurred in R. microocellata and R. undulata. Phocanema spp. was first recorded in L. fullonica, L. naevus, R. microocellata, R. montagui, and R. undulata, as Proleptus sp. in L. fullonica, and A. simplex in L. fullonica and R. clavata, Rockacestus sp. and Nybelinia sp. in R. undulata, and gill-myxosporeans on L. fullonica, L. naevus, R. microocellata, and R. undulata. The occurrence of 16 new host-parasite associations suggests potential environmental changes. Information provided by trophically transmitted helminths confirmed an opportunistic skate diet based on crustaceans and fish. We discuss results in terms of host fitness loss, bioindicator role of parasites, and anisakiasis risk. We recommend incorporating parasitology in research to improve elasmobranch conservation.

Chemical detection triggers honey bee defense against a destructive parasitic threat.

Invasive species events related to globalization are increasing, resulting in parasitic outbreaks. Understanding of host defense mechanisms is needed to predict and mitigate against the consequences of parasite invasion. Using the honey bee Apis mellifera and the mite Varroa destructor, as a host-parasite model, we provide a comprehensive study of a mechanism of parasite detection that triggers a behavioral defense associated with social immunity. Six Varroa-parasitization-specific (VPS) compounds are identified that (1) trigger Varroa-sensitive hygiene (VSH, bees' key defense against Varroa sp.), (2) enable the selective recognition of a parasitized brood and (3) induce responses that mimic intrinsic VSH activity in bee colonies. We also show that individuals engaged in VSH exhibit a unique ability to discriminate VPS compounds from healthy brood signals. These findings enhance our understanding of a critical mechanism of host defense against parasites, and have the potential to apply the integration of pest management in the beekeeping sector.

Plant defense resistance in natural enemies of a specialist insect herbivore.

Plants defend themselves against herbivores through the production of toxic and deterrent metabolites. Adapted herbivores can tolerate and sometimes sequester these metabolites, allowing them to feed on defended plants and become toxic to their own enemies. Can herbivore natural enemies overcome sequestered plant defense metabolites to prey on adapted herbivores? To address this question, we studied how entomopathogenic nematodes cope with benzoxazinoid defense metabolites that are produced by grasses and sequestered by a specialist maize herbivore, the western corn rootworm. We find that nematodes from US maize fields in regions in which the western corn rootworm was present over the last 50 y are behaviorally and metabolically resistant to sequestered benzoxazinoids and more infective toward the western corn rootworm than nematodes from other parts of the world. Exposure of a benzoxazinoid-susceptible nematode strain to the western corn rootworm for 5 generations results in higher behavioral and metabolic resistance and benzoxazinoid-dependent infectivity toward the western corn rootworm. Thus, herbivores that are exposed to a plant defense sequestering herbivore can evolve both behavioral and metabolic resistance to plant defense metabolites, and these traits are associated with higher infectivity toward a defense sequestering herbivore. We conclude that plant defense metabolites that are transferred through adapted herbivores may result in the evolution of resistance in herbivore natural enemies. Our study also identifies plant defense resistance as a potential target for the improvement of biological control agents.

Challenging the Preference-Performance Hypothesis in an above-belowground insect.

The relationship between female oviposition preference and offspring performance has been a question of special interest in the study of host plant selection by phytophagous insects. The Preference-Performance Hypothesis (PPH) is one of the main hypotheses proposed to explain this relationship, stating that females should preferentially lay eggs on plants providing the best larval development. The PPH has been extensively tested on aboveground insects but its application to species with belowground larvae is still mostly unknown. In this study, the PPH was quantitatively tested in an above-belowground insect, the cabbage root fly Delia radicum. Female oviposition preference and larval performance were estimated on three brassicaceous species (Brassica oleracea, Brassica rapa, and Sinapis alba) as well as between four cultivars of B. rapa and four cultivars of S. alba. Larval performance was estimated through their survival and through three life-history traits (LHT) of emerging adults. The PPH was supported at the intraspecific scale but only in B. rapa and for some, but not all, of the life-history traits. No support for the PPH was found in S. alba as well as at the interspecific scale. This study pleads for the integration of insects with both above- and belowground life stages in the preference-performance debate. Moreover, it raises the importance of measuring several variables to estimate larval performance and to test the PPH quantitatively, both at the plant intraspecific and interspecific scales, before drawing general conclusions.

Impact of Seasonal and Temperature-Dependent Variation in Root Defense Metabolites on Herbivore Preference in Taraxacum officinale.

Plants experience seasonal fluctuations in abiotic and biotic factors such as herbivore attack rates. If and how root defense expression co-varies with seasonal fluctuations in abiotic factors and root herbivore attack rates is not well understood. Here, we evaluated seasonal changes in defensive root latex chemistry of Taraxacum officinale plants in the field and correlated the changes with seasonal fluctuations in abiotic factors and damage potential by Melolontha melolontha, a major natural enemy of T. officinale. We then explored the causality and consequences of these relationships under controlled conditions. The concentration of the defensive sesquiterpene lactone taraxinic acid ß-D glucopyranosyl ester (TA-G) varied substantially over the year and was most strongly correlated to mean monthly temperature. Both temperature and TA-G levels were correlated with annual fluctuations in potential M. melolontha damage. Under controlled conditions, plants grown under high temperature produced more TA-G and were less attractive for M. melolontha. However, temperature-dependent M. melolontha feeding preferences were not significantly altered in TA-G deficient transgenic lines. Our results suggest that fluctuations in temperature leads to variation in the production of a root defensive metabolites that co-varies with expected attack of a major root herbivore. Temperature-dependent herbivore preference, however, is likely to be modulated by other phenotypic alterations.

Distinct defense strategies allow different grassland species to cope with root herbivore attack.

Root-feeding insect herbivores are of substantial evolutionary, ecological and economical importance. Plants defend themselves against insect herbivores through a variety of tolerance and resistance strategies. To date, few studies have systematically assessed the prevalence and importance of these strategies for root-herbivore interactions across different plant species. Here, we characterize the defense strategies used by three different grassland species to cope with a generalist root herbivore, the larvae of the European cockchafer Melolontha melolontha. Our results reveal that the different plant species rely on distinct sets of defense strategies. The spotted knapweed (Centaurea stoebe) resists attack by dissuading the larvae through the release of repellent chemicals. White clover (Trifolium repens) does not repel the herbivore, but reduces feeding, most likely through structural defenses and low nutritional quality. Finally, the common dandelion (Taraxacum officinale) allows M. melolontha to feed abundantly but compensates for tissue loss through induced regrowth. Thus, three co-occurring plant species have evolved different solutions to defend themselves against attack by a generalist root herbivore. The different root defense strategies may reflect distinct defense syndromes.

Multivariate Analysis of Multiple Datasets: a Practical Guide for Chemical Ecology.

Chemical ecology has strong links with metabolomics, the large-scale study of all metabolites detectable in a biological sample. Consequently, chemical ecologists are often challenged by the statistical analyses of such large datasets. This holds especially true when the purpose is to integrate multiple datasets to obtain a holistic view and a better understanding of a biological system under study. The present article provides a comprehensive resource to analyze such complex datasets using multivariate methods. It starts from the necessary pre-treatment of data including data transformations and distance calculations, to the application of both gold standard and novel multivariate methods for the integration of different omics data. We illustrate the process of analysis along with detailed results interpretations for six issues representative of the different types of biological questions encountered by chemical ecologists. We provide the necessary knowledge and tools with reproducible R codes and chemical-ecological datasets to practice and teach multivariate methods.

A mechanism for sequence specificity in plant-mediated interactions between herbivores.

Herbivore communities are shaped by indirect plant-mediated interactions whose outcomes are strongly dependent on the sequence of herbivore arrival. However, the mechanisms underlying sequence specificity are poorly understood. We examined the mechanisms that govern sequence-specific effects of the interaction between two specialist maize herbivores, the leaf feeder Spodoptera frugiperda and the root feeder Diabrotica virgifera virgifera. In the field, S. frugiperda reduces D. v. virgifera abundance, but only when it arrives on the plant first. In behavioral experiments, D. v. virgifera larvae continued feeding on plants that they had infested before leaf infestation, but refused to initiate feeding on plants that were infested by S. frugiperda before their arrival. Changes in root-emitted volatiles were sufficient to elicit this sequence-specific behavior. Root volatile and headspace mixing experiments showed that early-arriving D. v. virgifera larvae suppressed S. frugiperda-induced volatile repellents, which led to the maintenance of host attractiveness to D. v. virgifera. Our study provides a physiological and behavioral mechanism for sequence specificity in plant-mediated interactions and suggests that physiological canalization of behaviorally active metabolites can drive sequence specificity and result in strongly diverging herbivore distribution patterns.

Metazoan parasite communities in Alosa alosa (Linnaeus, 1758) and Alosa fallax (Lacépède, 1803) (Clupeidae) from North-East Atlantic coastal waters and connected rivers.

Metazoan parasites were studied in 96 Alosa alosa and 78 Alosa fallax from North-East Atlantic coastal waters and connected rivers (among them three sympatric sites) in order to increase knowledge on these anadromous endangered fish and measure the parasitic impact on host condition. All shads were infected by one to six metazoan parasite taxa among the 12 identified in the whole sampling, with a mean abundance of parasites higher for A. alosa (167 ± 10) than for A. fallax (112 ± 11). Helminths, mostly trophically transmitted, were the best represented (eight taxa, prevalence up to 99%) in contrast with crustaceans and Petromyzontidae that rarely occurred (four taxa, prevalence <6%). Despite some quantitative differences, metazoan parasite communities of A. alosa and A. fallax remained stable in composition whatever the host developmental stage, sex, sample site, and salinity. Among the nine parasite taxa harbored by each Alosa species, six were shared with some differences in distribution patterns including in sympatric conditions, suggesting increasing dissimilarities between A. alosa and A. fallax with the age. Information on feeding ecology provided by trophically transmitted helminths confirmed euryphagous opportunistic diet of immatures and adults of both shad species, and assessed feeding of adults during spawning migrations. Our study also revealed the significant negative impact of Hemiurus appendiculatus on A. alosa and Pronoprymna ventricosa on A. fallax. Because helminth parasites are omnipresent in the shads and decrease their fitness, parasitological data must be included in further investigations and management programs on A. alosa and A. fallax.

Activation of peripheral blood mononuclear cells by dengue virus infection depotentiates balapiravir.

In a recent clinical trial, balapiravir, a prodrug of a cytidine analog (R1479), failed to achieve efficacy (reducing viremia after treatment) in dengue patients, although the plasma trough concentration of R1479 remained above the 50% effective concentration (EC(50)). Here, we report experimental evidence to explain the discrepancy between the in vitro and in vivo results and its implication for drug development. R1479 lost its potency by 125-fold when balapiravir was used to treat primary human peripheral blood mononuclear cells (PBMCs; one of the major cells targeted for viral replication) that were preinfected with dengue virus. The elevated EC(50) was greater than the plasma trough concentration of R1479 observed in dengue patients treated with balapiravir and could possibly explain the efficacy failure. Mechanistically, dengue virus infection triggered PBMCs to generate cytokines, which decreased their efficiency of conversion of R1479 to its triphosphate form (the active antiviral ingredient), resulting in decreased antiviral potency. In contrast to the cytidine-based compound R1479, the potency of an adenosine-based inhibitor of dengue virus (NITD008) was much less affected. Taken together, our results demonstrate that viral infection in patients before treatment could significantly affect the conversion of the prodrug to its active form; such an effect should be calculated when estimating the dose efficacious for humans.

Manipulating feeding stimulation to protect crops against insect pests?

Enhancing natural mechanisms of plant defense against herbivores is one of the possible strategies to protect cultivated species against insect pests. Host plant feeding stimulation, which results from phagostimulant and phagodeterrent effects of both primary and secondary metabolites, could play a key role in levels of damage caused to crop plants. We tested this hypothesis by comparing the feeding intensity of the pollen beetle Meligethes aeneus on six oilseed rape (Brassica napus) genotypes in a feeding experiment, and by assessing the content of possible phagostimulant and phagodeterrent compounds in tissues targeted by the insect (flower buds). For this purpose, several dozens of primary and secondary metabolites were quantified by a set of chromatographic techniques. Intergenotypic variability was found both in the feeding experiment and in the metabolic profile of plant tissues. Biochemical composition of the perianth was in particular highly correlated with insect damage. Only a few compounds explained this correlation, among which was sucrose, known to be highly phagostimulating. Further testing is needed to validate the suggested impact of the specific compounds we have identified. Nevertheless, our results open the way for a crop protection strategy based on artificial selection of key determinants of insect feeding stimulation.

Understanding crop colonization of oilseed rape crops by the cabbage stem flea beetle (Psylliodes chrysocephala L. (Coleoptera: Chrysomelidae)).

BACKGROUND: Development of accurate pest monitoring systems is essential for the establishment of integrated pest management strategies. Information about the pest behavior during the colonization process, as well as the sex and reproductive status of the colonizing population often are lacking and hinder their development. The cabbage stem flea beetle (CSFB, Psylliodes chrysocephala) can cause the complete destruction of oilseed rape crops (OSR, Brassica napus). In the present study, the colonization process of OSR fields by the CSFB was studied. RESULTS: More individuals were caught on the outward facing side of the traps than the side of the trap facing towards the crop at the field border and catches were higher on the trapping units at the center of the field than at its border, suggesting that more beetles were entering than leaving the crop. Catches were higher on lower traps placed near to the crop than on those positioned further from the ground and also were higher during the day than late afternoon and night. The sex-ratio of individuals caught was skewed towards males and sexual maturity was acquired for females during the experiment. Integration of sampling data with local meteorological data showed that the catches correlated mostly with air temperature and relative humidity. CONCLUSION: This study provides new information about the dispersion of the CSFB in OSR fields during the colonization process, and highlights correlations between local meteorological factors and activity of the CSFB, and represent a new step towards implementing monitoring strategies against this pest. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Potential Complementary Effect of Zinc and Alkalihalobacillus clausii on Gut Health and Immunity: A Narrative Review.

A balanced microbiota-microorganisms that live in the gut-is crucial in the early years of a child's life, while dysbiosis-altered microbiota-has been linked to the development of various diseases. Probiotics, such as Alkalihalobacillus clausii, are commonly used to restore the balance of gut microbiota and have shown additional antimicrobial and immunomodulatory properties. Intake of micronutrients can affect the structure and function of the gut barrier and of the microbiota by having multiple effects on cellular metabolism (e.g., immunomodulation, gene expression, and support structure proteins). An inadequate zinc intake increases the risk of deficiency and associated immune dysfunctions; it is responsible for an increased risk of developing gastrointestinal diseases, respiratory infections, and stunting. Paediatric zinc deficiency is a public health concern in many countries, especially in low-income areas. Currently, zinc supplementation is used to treat childhood diarrhoea. This review examines how combining A. clausii and zinc could improve dysbiosis, gut health, and immunity. It suggests that this combination could be used to prevent and treat infectious diseases and diarrhoea in children up to adolescence.

Urinary VOCs as biomarkers of early stage lung tumour development in mice.

BACKGROUND: Lung cancer is the primary cause of cancer-induced death. In addition to prevention and improved treatment, it has increasingly been established that early detection is critical to successful remission. OBJECTIVE: The aim of this study was to identify volatile organic compounds (VOCs) in urine that could help diagnose mouse lung cancer at an early stage of its development. METHODS: We analysed the VOC composition of urine in a genetically engineered lung adenocarcinoma mouse model with oncogenic EGFR doxycycline-inducible lung-specific expression. We compared the urinary VOCs of 10 cancerous mice and 10 healthy mice (controls) before and after doxycycline induction, every two weeks for 12 weeks, until full-blown carcinomas appeared. We used SPME fibres and gas chromatography - mass spectrometry to detect variations in cancer-related urinary VOCs over time. RESULTS: This study allowed us to identify eight diagnostic biomarkers that help discriminate early stages of cancer tumour development (i.e., before MRI imaging techniques could identify it). CONCLUSION: The analysis of mice urinary VOCs have shown that cancer can induce changes in odour profiles at an early stage of cancer development, opening a promising avenue for early diagnosis of lung cancer in other models.

CD40 ligand and MHC class II expression are essential for human peripheral B cell tolerance.

Hyper-IgM (HIGM) syndromes are primary immunodeficiencies characterized by defects of class switch recombination and somatic hypermutation. HIGM patients who carry mutations in the CD40-ligand (CD40L) gene expressed by CD4(+) T cells suffer from recurrent infections and often develop autoimmune disorders. To investigate the impact of CD40L-CD40 interactions on human B cell tolerance, we tested by ELISA the reactivity of recombinant antibodies isolated from single B cells from three CD40L-deficient patients. Antibody characteristics and reactivity from CD40L-deficient new emigrant B cells were similar to those from healthy donors, suggesting that CD40L-CD40 interactions do not regulate central B cell tolerance. In contrast, mature naive B cells from CD40L-deficient patients expressed a high proportion of autoreactive antibodies, including antinuclear antibodies. Thus, CD40L-CD40 interactions are essential for peripheral B cell tolerance. In addition, a patient with the bare lymphocyte syndrome who could not express MHC class II molecules failed to counterselect autoreactive mature naive B cells, suggesting that peripheral B cell tolerance also depends on major histocompatibility complex (MHC) class II-T cell receptor (TCR) interactions. The decreased frequency of MHC class II-restricted CD4(+) regulatory T cells in CD40L-deficient patients suggests that these T cells may mediate peripheral B cell tolerance through CD40L-CD40 and MHC class II-TCR interactions.

Pain management recommendations during the progression of SARS-CoV-2 infection.

COVID-19, an infection caused by SARS-CoV-2, had a devastating impact on people's lives. The pandemic placed a heavy burden on healthcare systems and impacted the care of patients, including those with pain. This narrative review aims to highlight the challenges in managing pain and fever resulting from COVID-19 and pre-existing conditions, and to discuss the role of over the counter analgesics as a key part of the COVID-19 treatment regimen. As most patients with COVID-19 are being managed in the outpatient setting, lifestyle interventions and over the counter analgesics are readily available options to effectively treat pain and fever, which can help to decrease the burden on the healthcare system during the COVID-19 pandemic.

COVID-19 is an infection caused by SARS-CoV-2. The COVID-19 pandemic not only affects patient lives, but also heavily impacts healthcare systems. This review aims to discuss the available literature on how to manage pain from COVID-19 and encourage a consensus meeting for recommendations. As most patients with COVID-19 are being managed in the outpatient setting, lifestyle interventions and over the counter analgesics are readily available options to effectively treat pain and fever, which can help to decrease the demand on the healthcare system during the COVID-19 pandemic.

Age-specific allocation of glucosinolates within plant reproductive tissues.

The Optimal Defense Theory (ODT) predicts that the distribution of defenses within a plant should mirror the value and vulnerability of each tissue. Although the ODT has received much experimental support, very few studies have examined defense allocation among reproductive tissues and none assessed simultaneously how these defenses evolve with age. We quantified glucosinolates in perianths, anthers and pistils at different bud maturity stages (i.e., intermediate flower buds, old flower buds and flowers) of undamaged and mechanically damaged plants of an annual brassicaceous species. The youngest leaf was used as a reference for vegetative organs, since it is predicted to be one of the most defended. In line with ODT predictions, reproductive tissues were more defended than vegetative tissues constitutively, and within the former, pistils and anthers more defended than perianths. No change in the overall defense level was found between bud maturity stages, but a significant temporal shift was observed between pistils and anthers. Contrary to ODT predictions, mechanical damage did not induce systemic defenses in leaves but only in pistils. Our results show that defense allocation in plant reproductive tissues occurs at fine spatial and temporal scales, extending the application framework of the ODT. They also demonstrate interactions between space and time in fine-scale defense allocation.

Sex- and maturity-dependent antennal detection of host plant volatiles in the cabbage root fly, Delia radicum.

Adult insect behaviour in response to plant-emitted volatile compounds varies between the sexes and as a function of maturity. These differences in behavioural responses can be due to modulation in the peripheral or central nervous system. In the cabbage root fly, Delia radicum, behavioural effects of certain host plant volatiles on mature female behaviour have been evaluated, and a large number of compounds emitted by brassicaceous host plants have been identified. We recorded here dose-dependent electroantennogram responses to all tested compounds and investigated if the antennal detection of individual volatile compounds emitted by intact and damaged host plants differs between male and female, as well as immature and mature flies. Our results showed dose-dependent responses in mature and immature males and females. Mean response amplitudes varied significantly between sexes for three compounds, and between maturity states for six compounds. For some additional compounds significant differences occurred only for high stimulus doses (interaction between dose and sex and/or dose and maturity status). Multivariate analysis revealed a significant global effect of maturity on electroantennogram response amplitudes and for one experimental session also a significant global effect of the sex. Interestingly, allyl isothiocyanate, a compound stimulating oviposition behaviour, elicited stronger responses in mature than in immature flies, whereas ethylacetophenone, an attractive flower volatile, elicited stronger responses in immature than in mature flies, which correlates with the behavioural role of these compounds. Several host-derived compounds elicited stronger responses in females than in males and, at least at high doses, stronger responses in mature than in immature flies, indicating differential antennal sensitivity to behaviourally active compounds. Six compounds did not cause any significant differences in responses between the different groups of flies. Our results thus confirm peripheral plasticity in plant volatile detection in the cabbage root fly and provide a basis for future behavioural investigations on the function of individual plant compounds.

Potential complementary and/or synergistic effects of curcumin and boswellic acids for management of osteoarthritis.

For several thousand years (~4000) Boswellia serrata and Curcuma longa have been used in Aryuvedic medicine for treatment of various illnesses, including asthma, peptic ulcers, and rheumatoid arthritis, all of which are mediated through pathways associated with inflammation and pain. Although the in vivo pharmacology of both these natural ingredients is difficult to study because of poor bioavailability, in vitro data suggest that both influence gene expression mediated through nuclear factor kappa B (NF-κB). Therefore, the activity of pathways associated with inflammation (including NF-κB and lipoxygenase- and cyclooxygenase-mediated reduction in leukotrienes/prostaglandins) and those involved in matrix degradation and apoptosis are reduced, resulting in a reduction in pain. Additive activity of boswellic acids and curcumin was observed in preclinical models and synergism was suggested in clinical trials for the management of osteoarthritis (OA) pain. Overall, studies of these natural ingredients, alone or in combination, revealed that these extracts relieved pain from OA and other inflammatory conditions. This may present an opportunity to improve patient care by offering alternatives for patients and physicians, and potentially reducing nonsteroidal anti-inflammatory or other pharmacologic agent use. Additional research is needed on the effects of curcumin on the microbiome and the influence of intestinal metabolism on the activity of curcuminoids to further enhance formulations to ensure sufficient anti-inflammatory and antinociceptive activity. This narrative review includes evidence from in vitro and preclinical studies, and clinical trials that have evaluated the mechanism of action, pharmacokinetics, efficacy, and safety of curcumin and boswellic acids individually and in combination for the management of OA pain.

A protocol for increased throughput phenotyping of plant resistance to the pollen beetle.

BACKGROUND: Improving crop resistance to insect herbivores is a major research objective in breeding programs. Although genomic technologies have increased the speed at which large populations can be genotyped, breeding programs still suffer from phenotyping constraints. The pollen beetle (Brassicogethes aeneus) is a major pest of oilseed rape for which no resistant cultivar is available to date, but previous studies have highlighted the potential of white mustard as a source of resistance and introgression of this resistance appears to be a promising strategy. Here we present a phenotyping protocol allowing mid-throughput (i.e., increased throughput compared to current methods) acquisition of resistance data, which could then be used for genetic mapping of QTLs. RESULTS: Contrasted white mustard genotypes were selected from an initial field screening and then evaluated for their resistance under controlled conditions using a standard phenotyping method on entire plants. We then upgraded this protocol for mid-throughput phenotyping, by testing two alternative methods. We found that phenotyping on detached buds did not provide the same resistance contrasts as observed with the standard protocol, in contrast to the phenotyping protocol with miniaturized plants. This protocol was then tested on a large panel composed of hundreds of plants. A significant variation in resistance among genotypes was observed, which validates the large-scale application of this new phenotyping protocol. CONCLUSION: The combination of this mid-throughput phenotyping protocol and white mustard as a source of resistance against the pollen beetle offers a promising avenue for breeding programs aiming to improve oilseed rape resistance. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.