Control Of Mites Research Articles

Molecular Basis for the Selectivity of the Succinate Dehydrogenase Inhibitor Cyflumetofen between Pest and Predatory Mites.

Acaricides that act as inhibitors of the mitochondrial succinate dehydrogenase (SDHIs) provide excellent control of phytophagous mites but display limited toxicity to predatory mites and other beneficial organisms. However, the molecular mechanism of selectivity is not fully understood. Here, we first confirm that SDHI acaricides are over 10,000-fold more toxic to spider mites than predatory mites. Next, we show that differential penetration, pro-acaricide activation, or metabolism are most likely not the main reason for this selectivity. In contrast, the inhibition of AB-1 on the SDH target is approximately 200-fold more potent in spider mites compared to predatory mites, revealing strong target-site selectivity. Strikingly, a key motif associated with differential binding was identified and validated by gene editing in Drosophila. Our findings contribute to understanding the selectivity of SDHIs, which can be used for the rational design of selective acaricides in support of an integrated pest management.

Recognition of spider mite infestations in jujube trees based on spectral-spatial clustering of hyperspectral images from UAVs.

Spider mite infestations are a serious hazard for jujube trees in China. The use of remote sensing technology to evaluate the health of jujube trees in large-scale intensive agricultural production is an effective means of agricultural control. Hyperspectral remote sensing has a higher spectral resolution and richer spectral information than conventional multispectral remote sensing, which improves the detection of crop pests and diseases. We used hyperspectral remote sensing data from jujube fields infested with spider mite in Hotan Prefecture, Xinjiang to evaluate their use in monitoring this important pest. We fused spectral and spatial information from the hyperspectral images and propose a method of recognizing spider mite infestations of jujube trees. Our method is based on the construction of spectral features, the fusion of spatial information and clustering of these spectral-spatial features. We evaluated the effect of different spectral-spatial features and different clustering methods on the recognition of spider mite in jujube trees. The experimental results showed that the overall accuracy of the method for the recognition of spider mites was >93% and the overall accuracy of the band clustering-density peak clustering model for the recognition of spider mite reached 96.13%. This method can be applied to the control of jujube spider mites in agricultural production.

Effects of far-red light on tritrophic interactions between the two-spotted spider mite (Tetranychus urticae) and the predatory mite Phytoseiulus persimilis on tomato.

The use of light-emitting diode (LED) lights in horticulture allows growers to adjust the light spectrum to optimize crop production and quality. However, changes in light quality can also influence plant-arthropod interactions, with possible consequences for pest management. The addition of far-red light has been shown to interfere with plant immunity, thereby increasing plant susceptibility to biotic stress and increasing pest performance. Far-red light also influences plant emission of volatile organic compounds (VOCs) and might thus influence tritrophic interactions with biological control agents. We investigated how far-red light influences the VOC-mediated attraction of the predatory mite Phytoseiulus persimilis to tomato plants infested with Tetranychus urticae, and its ability to control T. urticae populations. Far-red light significantly influences herbivore-induced VOC emissions of tomato plants, characterized by a change in relative abundance of terpenoids, but this did not influence the attraction of P. persimilis to herbivore-induced plants. Supplemental far-red light led to an increased population growth of T. urticae and increased numbers of P. persimilis. This resulted in a stronger suppression of T. urticae populations under supplemental far-red light, to similar T. urticae numbers as in control conditions without supplemental far-red light. We conclude that supplemental far-red light can change herbivore-induced VOC emissions but does not interfere with the attraction of the predator P. persimilis. Moreover, far-red light stimulates biological control of spider mites in glasshouse tomatoes due to increased population build-up of the biocontrol agent. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Control of brown mites in apple with miticides, 2020

Control of brown mites in apple with miticides, 2020

Efficacy of insecticides for control of two-spotted spider mites in strawberries, 2022

Efficacy of insecticides for control of two-spotted spider mites in strawberries, 2022

Control of brown mites in apple with miticides, 2021

Control of brown mites in apple with miticides, 2021

Potential of ferritin 2 as an antigen for the development of a universal vaccine for avian mites, poultry red mites, tropical fowl mites, and northern fowl mites.

Poultry red mites (PRMs, Dermanyssus gallinae), blood-sucking ectoparasites, are a threat to the poultry industry because of reduced production caused by infestation. In addition, tropical fowl mites (TFMs, Ornithonyssus bursa) and northern fowl mites (NFMs, Ornithonyssus sylviarum) are hematophagous, distributed in various regions, genetically and morphologically close to PRMs, and cause similar problems to the poultry industry. Vaccine approaches have been studied for PRM control, and several molecules have been identified in PRMs as candidates for effective vaccine antigens. The development of an anti-PRM vaccine as a universal vaccine with broad efficacy against avian mites could improve the productivity of poultry farms worldwide. Molecules that are highly conserved among avian mites and have critical functions in the physiology and growth of mites could be ideal antigen candidates for the development of universal vaccines. Ferritin 2 (FER2), an iron-binding protein, is critical for the reproduction and survival of PRMs and has been reported as a useful vaccine antigen for the control of PRMs and a candidate for the universal vaccine antigen in some tick species. Herein, we identified and characterized FER2 in TFMs and NFM. Compared with the sequence of PRM, the ferroxidase centers of the heavy chain subunits were conserved in FER2 of TFMs and NFMs. Phylogenetic analysis revealed that FER2 belongs to clusters of secretory ferritins of mites and other arthropods. Recombinant FER2 (rFER2) proteins from PRMs, TFMs, and NFMs exhibited iron-binding abilities. Immunization with each rFER2 induced strong antibody responses in chickens, and each immune plasma cross-reacted with rFER2 from different mites. Moreover, mortality rates of PRMs fed with immune plasma against rFER2 from TFMs or NFMs, in addition to PRMs, were higher than those of control plasma. rFER2 from each avian mite exhibited anti-PRM effects. This data suggests that it has the potential to be used as an antigen candidate for a universal vaccine against avian mites. Further studies are needed to access the usefulness of FER2 as a universal vaccine for the control of avian mites.

The Role of Spray Pattern and Operating Pressure and their Interactions on the Control of Tetranychus urticae Koch. in Eggplant Plants Under Greenhouse Conditions

Ibraheem, S.N. and M.H. Alheidary. 2023. The Role of Spray Pattern and Operating Pressure and their Interactions on the Control of Tetranychus urticae Koch. in Eggplant Plants Under Greenhouse Conditions. Arab Journal of Plant Protection, 41(2): 105-113. https://doi.org/10.22268/AJPP-41.2.105113 The mite Tetranychus urticae Koch is one of the most dangerous pests that attack the eggplant (Solanum melongena L.). Several attempts were carried out for controlling this pest to avoid crop losses through the extensive use of chemical products. The present study aimed to shed light on the effect of spraying pattern used coupled with selecting the appropriate operating pressure to obtain a suitable droplet size and density deposited on the leaves to get optimal effect for the control of mites which attack eggplant under greenhouse conditions. Six types of spray patterns (from top, side, bottom, top and side together, bottom and side together, and combination spray mode) toward plant canopy and two different operating pressures (2 and 5 bar) were tested. The factorial experiment was applied in split-plot arrangement in a complete random design. The significant differences between the parameters studied were tested by calculating the least significant difference (L.S.D.) at P=0.05. The findings showed a clear significant influence in the control efficiency by both spraying patterns and operating pressures. Significant mortality efficiency was observed using the bottom spray pattern from the plant canopy especially at 5 bar constant pressure in comparison with the other spray patterns at the same operating pressure. It was also observed that spraying with two-nozzle spray pattern from the bottom led to an increase in the efficiency of killing mites not only on the bottom part of the plant but also those present in the upper part of the plant, with a morality rate of 22.67% compared to the control treatment. The results also illustrated that the use of bottom spray pattern reduced the amount of pesticide applied by more than 46% in comparison to the other treatments. Keywords: sprayer, nozzle, pesticide, spray process characteristics, pest control, morality efficiency

Volatile compounds from soybeans under multiple on herbivores infestations attract the predatory mite Neoseiulus californicus (Acari: Phytoseiidae).

Plant-induced resistance can be an important component of soybean mites biological control programs. This work evaluates the preference of predatory mite Neoseiulus californicus (Acari: Phytoseiidae) to soybean plants under single and multiple herbivory conditions by two-spotted spider mite Tetranychus urticae (Acari: Tetranychidae), and velvetbean caterpillar Anticarsia gemmatalis (Lepidoptera: Noctuidae). Using a Y olfactometer, the following scenarios were evaluated: soybean with no infestation and soybean infested with A. gemmatalis; soybean infested with T. urticae and A. gemmatalis, and soybean infested with T. urticae and with both T. urticae and A. gemmatalis. Volatile compounds released by plants were analyzed and identified by a Trace GC Ultra gas chromatograph coupled to a mass spectrometer with a solid phase micro-extraction ion-trap. The predatory mite N. californicus preferred soybean plants infested with T. urticae compared to those infested with A. gemmatalis. Multiple infestation did not interfere with its preference to T. urticae. Multiple herbivory of T. urticae and A. gemmatalis modified the chemical profile of volatile compounds emitted by soybean plants. However, it did not interfere with the search behavior of N. californicus. Out of the 29 identified compounds only five promoted predatory mite response. Thus, regardless of single or multiple herbivory by T. urticae with or without A. gemmatalis, the indirect induced resistance mechanisms operate similarly. As such, this mechanism contributes to an increase in the encounter rate between predator and prey for N. Californicus and T. urticae, and the efficacy of biological control of mites on soybean.

Transcriptome analysis revealed detoxification gene expression changes in Tetranychus cinnabarinus challenged with ethyl oleate.

Natural acaricides are potential biorational mite control alternatives to conventional chemical acaricides. However, little is known about the molecular mechanism of defense response to natural acaricides in mites. We previously reported significant acaricidal properties of ethyl oleate (EO) against Tetranychus cinnabarinus (here referred to as a sibling species of two-spotted spider mite, Tetranychus urticae), a highly polyphagous pest devastating crops in fields and greenhouses worldwide. In this study, we explored the molecular responses of T. cinnabarinus exposed to EO using RNA-Seq and differentially expressed gene (DEG) analysis. A total of 131, 185, and 154 DEGs were identified in T. cinnabarinus after 1, 6, and 24h of EO treatment. In addition, 36 putative detoxification-related DEGs, including 10 cytochrome P450s (P450s), three glutathione S-transferases (GSTs), nine UDP-glycosyltransferases (UGTs), eight esterases (ESTs), and six ATP-binding cassette transporters (ABC transporters), were identified. Interestingly, the upregulation of these detoxification-related genes might be the main defense response of T. cinnabarinus exposed to EO. A quantitative real-time PCR analysis indicated that the expression profiles of 19 random DEGs were consistent with the RNA-Seq results. These findings serve as valuable information for a better understanding of the acaricide-mite interaction and molecular mechanisms involved in the defense response of T. cinnabarinus against EO.

Sunflower-Associated Reductions in Varroa Mite Infestation of Honey Bee Colonies.

Landscapes can affect parasite epidemiology in wild and agricultural animals. Honey bees are threatened by loss of floral resources and by parasites, principally the mite Varroa destructor and the viruses it vectors. Existing mite control relies heavily on chemical treatments that can adversely affect bees. Alternative, pesticide-free control methods are needed to mitigate infestation with these ectoparasites. Many flowering plants provide nectar and pollen that confer resistance to parasites. Enrichment of landscapes with antiparasitic floral resources could therefore provide a sustainable means of parasite control in pollinators. Floral rewards of Asteraceae plants can reduce parasitic infection in diverse bee species, including honey and bumble bees. Here, we tested the effects of sunflower (Helianthus annuus) cropland and pollen supplementation on honey bee resistance to macro- and microparasites. Although sunflower had nonsignificant effects on microparasites, We found that increased sunflower pollen availability correlated with reduced Varroa mite infestation in landscapes and pollen-supplemented colonies. At the landscape level, each doubling of sunflower crop area was associated with a 28% reduction in mite infestation. In field trials, late-summer supplementation of colonies with sunflower pollen reduced mite infestation by 2.75-fold relative to artificial pollen. United States sunflower crop acreage has declined by 2% per year since 1980, however, suggesting reduced availability of this floral resource. Although further research is needed to determine whether the observed effects represent direct inhibition of mite fecundity or mite-limiting reductions in honey bee brood-rearing, our findings suggest the potential for sunflower plantings or pollen supplements to counteract a major driver of honey bee losses worldwide.

Alternative food sources for Amblydromella caudiglans (Phytoseiidae) and effects on predation.

The addition of supplemental food sources for natural enemies is a growing component of conservation and augmentative biological control. Supplemental foods can be used to retain natural enemies when prey are scarce and to promote survival, fecundity, longevity, and development of natural enemy populations, especially generalist natural enemies. Amblydromella caudiglans (Schuster) (Acari: Phytoseiidae) is one of the most abundant predatory mites found in commercial apple orchards in Washington, USA, and contributes to spider mite control. However, because its widespread presence in commercial apple orchards was only recently discovered, how supplementary food sources affect its performance is unknown. In laboratory studies, we evaluated the performance (fecundity, retention, prey consumption) of the generalist phytoseiid A. caudiglans on commercially available supplemental food sources, including factitious prey (Ephestia eggs and Artemia brine shrimp cysts), and pollens of apple, pear, and cattail. We determined that A. caudiglans will not consume Ephestia eggs and performs best on cattail and pear pollens. Combinations of food sources did not enhance the performance of this predator compared to the best performing single-sources. The presence of alternative food sources did not decrease A. caudiglans predation of twospotted spider mite nymphs, except for Artemia brine shrimp cysts, which had a substantial handling time. These results lay the groundwork for identifying a way to promote and retain this natural enemy in tree fruit cropping systems through the use of food resource applications or floral plantings.

Identification of cassava germplasms resistant to two-spotted spider mite in China: From greenhouse large-scale screening to field validation.

Utilization of resistant germplasm is considered as an effective, economical and eco-friendly strategy for cassava pest management. Tetranychus urticae, known as the two-spotted spider mite (TSSM), is a devastating pest in Asian cassava planting countries as well as in China. However, the resistant levels of abundant cassava germplasms to TSSM remains largely unknown. To fill this knowledge gap, we conducted screening of 202 cassava germplasm for resistance to TSSM in China based on the classification of mite damage phenotype, under both greenhouse and field conditions. The three rounds of large-scale greenhouse experiments had identified two highly resistant (HR) varieties (C1115 and MIANDIAN), five resistant (R) varieties (SC5, SC9, SC15, COLUMBIA-4D and LIMIN) and five highly susceptible (HS) varieties (KU50, BREAD, SC205, TMS60444 and BRA900), besides, these 'HR' and 'R' varieties would significantly repress the normal development and reproduction of TSSM. In addition, the 12 cassava varieties selected from the greenhouse screening were further subjected to consecutive five years of field validation at Danzhou, Wuming and Baoshan. The seven resistant varieties not only exhibited stable TSSM-resistance performance across the three field environments, but also possessed the same resistant levels as the greenhouse identification, while the resistant varieties SC5 was an exception, which was identified as moderate resistant in Baoshan, indicating the variety-environment interaction may affect its resistance. Furthermore, regional yield estimation suggested that the higher the resistance level was, the better capacity in reducing the yield losses. This study demonstrated that the TSSM-resistant varieties could be considered as ideal materials in mite control or in future breeding programme of mite-resistant cassava plant.

The sublethal and transgenerational effects of pyridaben on fitness and induced tolerance in Panonychus citri

Panonychus citri is a dominant citrus pest mite in the world. Pesticide-induced population resurgence is a concern for environmental and resistance risk assessment in mite control. To evaluate the effective mite management strategies, the sublethal and transgenerational effects of pyridaben on P. citri were assessed using life-table parameters. Two segregated pyridaben-resistant and susceptible strains (previously obtained from the crossing between laboratory susceptible strain and pyridaben-resistant population) were used in our study. After exposure to LC30 pyridaben, the fecundity of Pyr_Control and Pyr_Rs strains were both significantly reduced in the parental generation (F0). Life-table results showed that LC30 pyridaben significantly induced the fecundity of both strains in F1 generation (egg stage was prolonged as well). Interestingly, this effect also stimulated the fecundity of F2 generation in Pyr_Control strain. But no significant effects were observed in Pyr_Rs strain. While detoxification enzyme assays indicated that only the activity of P450s was significantly activated by LC30 pyridaben in Pyr_Control and Pyr_Rs strains (1.242- fold and 1.355-fold increase, respectively). The expressions of P450 gene (CYP4CL2) and reproduction-related gene (Pc_Vg) were determined and the results showed that the expression of Pc_Vg of pyridaben treated mites in F0 generation was significantly reduced in both strains. In Pyr_Control and Pyr_Rs strains, the significant upregulation CYP4CL2 (2.029-fold and 2.516-fold increase, respectively) and PcVg (1.771-fold and 2.924-fold increase, respectively) of F1 generation suggested there were delayed hormesis effects on reproduction and developed tolerance to pyridaben. No significant difference was found between LC30 treatments and controls in F2 generation of both strains. These results provided evidence that transgenerational hormesis effects of low concentration of pyridaben might lead to mite population outbreak and resistance risk in the field through stimulating the reproduction and developing tolerance to pyridaben. Furthermore, LC30 pyridaben could affect the offspring populations of susceptible strain in the longer-term as compared to resistant strain.

Diversity, distribution and scope of utilization of phytoseiid mites (Acari: Phytoseiidae) in eastern India

The mites of the family Phytoseiidae are one of the most studied groups of predatory mites worldwide for its species diversity, wide distribution and feeding potential to act as biocontrol agents for control of pest mites and small soft bodied insect pests. Phytoseiid mites are boon of nature because they sustain on pests of many agri-horticultural crops which are indeed very difficult to manage relying solely on pesticides. Considering its immense importance, it has been extensively explored from eastern India to understand the inherent potential of these biocontrol agents. In the era of biological control, predatory mites, specifically those belonging to the family Phytoseiidae, are now appreciated with farmers worldwide as natural enemies for their exhaustive use in pest control in greenhouses and as well as in open field conditions.

Non-consumptive effects of predation risk on prey population regulations: empirical evidence from the invasive spider mite Tetranychus ludeni

Tetranychus ludeni Zacher is an invasive pest from Europe. It infests more than 300 plant species, including many economically important crops such as bean Phaseolus vulgaris L. (Fabales: Fabaceae), eggplant Solanum melongena L. (Solanales: Solanaceae), and many other cucurbitaceous plants. Gotoh et al. (2015) predict that this species could replace T. urticae to become a major crop pest in the world. It is often assumed that the success of biological control using predators is achieved through direct predation. However, it is still largely unknown whether predation risk could contribute to the effectiveness of prey biological control. In the present study we investigated the effects of cues from the predatory mite Phytoseiulus persimilis and killed T. ludeni on life history traits and offspring fitness of T. ludeni, providing novel knowledge for evaluation of the non-consumptive contribution to the biological control of T. ludeni and for future development of new spider mite control methods using these cues. We demonstrated that the cues from the predators and killed prey shortened the longevity by more than 23% and oviposition period by more than 35% and reduced the number of eggs laid by more than 31% in T. ludeni females. These cues also significantly reduced the intrinsic rate of increase and net population growth rate and prolonged the time to double the population size. Furthermore, in response to predator cues females significantly delayed their lifetime production of daughters and laid significantly smaller eggs. Offspring from predator-exposed mothers developed significantly more slowly. Our findings strongly suggest that the non-consumptive mortality caused by cues from both predators and killed prey can significantly reduce prey fitness, suppressing their population and lowering their damage to crops. The earlier prey death caused by these cues could result from the energic costs of natural enemy avoidance (Luong et al. 2017), decrease of foraging rate or food intake (Hermann & Thaler 2014) and increase of oxidative damage (Janssens & Stoks 2013). The present study partially supports previous reports that non-consumptive impacts of predation risk on prey are transgenerational (Tariel et al. 2020). Our study provides critical information for evaluation of biological control effectiveness using predators and paves the way for identification of chemical odors from the predator and killed prey and development of new materials and methods for the control of spider mite pests.

The advent of the mite omics era and integration of multiple technological approaches to mite control

Current food production involves not just the fundamental need to provide calories and proteins but full nutrition status under specific safety standards. Vegetable production, especially tomatoes, grown under highly intensive conditions and with cosmetic quality standards are exceedingly targeted by and susceptible to pests. Mites are mostly diminutive in size. Their widespread distribution and invasiveness (as new species) are often neglected, and they are commonly mistaken for similar known species, compounded by a limited understanding of their diverse biology and multitrophic interactions, resulting in pronounced crop losses due to late action.

Identification of guanine and hematin as arrestment pheromones of poultry red mites, Dermanyssus gallinae (Acari: Dermanyssidae) and their application in mite control

Dermanyssus gallinae, also known as the poultry red mite (PRM), is one of widespread ectoparasite in the poultry industry worldwide, causing direct and indirect detriments to poultry as well as substantial financial losses. Novel control methods are urgently needed to improve the current acaricide-based control of D. gallinae. The control approach based on arrestment pheromone is environment-friendly but the related research is limited in PRMs. In the present study we found two compounds from the mite feces acting as arrestment pheromones of D. gallinae, which could lead to mite arrestment upon contact. One is guanine, which was also found in unfed female mites’ acid-saline extract. The other is hematin. Moreover, it was found that the ferric ion of hematin played a pivotal role in stimulating the arrestment of mites. Finally, it was found the combination of guanine or/and hematin plus cypermethrin led to significantly improved mite-killing performance compared with cypermethrin, showing a promising potential of novel control method based on the arrestment pheromone.

Molecular verification of commercialized Neoseiulus californicus (McGregor) settlement before spider mite appearance in a Japanese pear orchard

A report of our earlier study described a phytoseiid mite Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) release system using a moisturized sheltered sachet for spider mite control. For the present study, we installed the predator release systems on 43 out of 71 trees in a Japanese pear orchard to survey the dispersal of N. californicus released from the predator release system. In the orchard, Japanese pear trees were trained on the overhead horizontal trellis system. Consequently, the trees were mutually connected through the trellis training system. We collected phytoseiid mites on the tree leaves and were subjected to PCR to examine their species composition. Results demonstrated that N. californicus were 62.37% of the total of 3,247 phytoseiid mites collected. Using microsatellite markers, commercialized N. californicus that had been released from the predator release systems were discriminated from indigenous N. californicus. Results demonstrated that phytoseiid mites consisting mainly of the commercialized N. californicus occurred one week earlier in trees with the predator release systems. Results also demonstrated that the commercialized N. californicus dispersal occurred mainly in trees installed with the predator release systems.

"Metabolomic diversity of local strains of Beauveria bassiana (Balsamo) Vuillemin and their efficacy against the cassava mite, Tetranychus truncatus Ehara (Acari: Tetranychidae)".

A desirable substitute for chemical pesticides is mycopesticides. In the current investigation, rDNA-ITS (Internal transcribed spacer) and TEF (Transcriptional Elongation Factor) sequencing were used for molecular identification of six Beauveria bassiana strains. Both, leaf discs and potted plant bioassaye were carried out to study their pathogenicity against the cassava mite, Tetranychus truncatus. LC50 and LC90 values of potential B. bassiana strains were estimated. We also discovered a correlation between intraspecific B. bassiana strains pathogenicity and comprehensive metabolome profiles. Bb5, Bb6, Bb8, Bb12, Bb15, and Bb21 strains were identified as B. bassiana by sequencing of rDNA-ITS and TEF segments and sequence comparison to NCBI (National Center for Biotechnology Information) GenBank. Out of the six strains tested for pathogenicity, Bb6, Bb12, and Bb15 strains outperformed against T. truncatus with LC50 values 1.4×106, 1.7×106, and 1.4×106 and with a LC90 values 7.3×107, 1.4×108, and 4.2×108 conidia/ml, respectively, at 3 days after inoculation and were considered as potential strains for effective mite control. Later, Gas Chromatography-Mass Spectrometry (GC-MS) analysis of the above six B. bassiana strains was done on secondary metabolites extracted with ethyl acetate revealed that the potential B. bassiana strains (Bb6, Bb12, and Bb15) have higher levels of acaricidal such as Bis(dimethylethyl)-phenol: Bb6 (5.79%), Bb12 (6.15%), and Bb15 (4.69%). Besides, insecticidal (n-Hexadecanoic acid), and insect innate immunity overcoming compound (Nonadecene) were also identified; therefore, the synergistic effect of these compounds might lead toa higher pathogenicity of B. bassiana against T. truncatus. Further, these compounds also exhibited two clusters, which separate the potential and non-potential strains in the dendrogram of Thin Layer Chromatography. These results clearly demonstrated the potentiality of the B. bassiana strains against T. truncatus due to the occurrence of their bioactive volatile metabolome.