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Vegetable and fruit contamination is recognized as a significant parasite transmission route. This review presents the current state of vegetables ad fruits contamination with food-borne parasitic protozoa worldwide. We consider the methodologies and strategies for detecting parasitic stages developed in the last decade and the contamination data. Asia had the highest number of reports (94 studies), followed by Africa (74 studies). At the country level, with 41 studies, Iran had the most reports among other countries, followed by Nigeria (28 studies). According to the studies included in the current review, 41.22% of vegetables and fruits were contaminated with different species of protozoan parasites. Among different continents, Asia accounted for the highest contamination rate of protozoan parasites (57.12%). Giardia spp. (10%) had the highest contamination rate in vegetables and fruits, followed by Entamoeba coli (8%), E. histolytica/dispar (7%), and Cryptosporidium spp. (6%). This study provides essential data for health authorities to develop food safety programs. The presence of protozoan parasites in fruits and vegetables highlights the critical need for maintaining rigorous food safety measures across the entire production and distribution process, particularly in countries that are major producers and distributors of these food items., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A complete three-dimensional reconstruction of the internal damage (oviposition holes, entry and exit galleries, cavities caused by fungal infection) of three destructive pests of olive fruit was obtained using micro-computed tomography. In the case of the olive fruit fly (Bactrocera oleae), complete reconstruction of the galleries was achieved. The galleries were colour-coded according to the size of the internal lumens produced by larval instars. In the case of the olive moth (Prays oleae), we confirmed that the larvae only consume olive stones, leaving pulp tissue intact. This study revealed the evolutionary defensive adaptation developed by larvae, creating entrance/exit gallery in the form of a zigzag with alternating angles to avoid the action of possible parasitoids. In the case of olive fruit rot, caused by fungal infection transmitted by the midge (Lasioptera berlesiana), microtomography revealed the infection cavity, which was delimited by a protective layer of tissue produced by the plant to isolate the infection zone, which contained fungal hyphae and reproductive organs of the fungus. Two ovoid cavities were observed below a single external orifice in the concave necrotic depression. These results were interpreted as successive ovipositions of B. oleae, followed by the parasitoid L. berlesiana. High-resolution 3D rendered images are included as well as supplementary videos that could be useful tools for future research and teaching aids., (© 2024. The Author(s).)

The genus Anastrepha contains some of the most important fruit pests in the Americas. It comprises more than 300 species, of which 129 occur in Brazil. The genus is divided into 26 species groups, including the pseudoparallela group with 31 species, whose known host plants are primarily fruits of the genus Passiflora (Passifloraceae). Fourteen species are recorded in Brazil. Here, a new species of Anastrepha reared from fruits of Passiflora actinia Hook. and Passiflora elegans Mast. from southern Brazil is described and illustrated. In addition, a synopsis of the Brazilian species of the pseudoparallela group is provided., (© 2024. Sociedade Entomológica do Brasil.)

The sliced and dried hawthorn berries are easily infested by insects during storage. This study aimed to determine the effect of insect infestation on the quality of hawthorn berries and assess the change at metabolite level by analyzing physicochemical property and metabolomics profiling. A total of 184 shared differential metabolites were obtained, mainly including flavonoids, fatty acids, carboxylic acids and derivatives, and nitrogenous compounds. Through receiver operating characteristic curve assessment, 9 significant differential markers were screened out to distinguish insect infestation of hawthorn berries. Correlation analysis showed that the color, total organic acids, total phenolics, and total flavonoids were effective indicators for quality evaluation of insect infestation, and uric acid and hippuric acid can serve as biomarkers for the quality deterioration of hawthorn berries during storage. This study demonstrated that insect infestation could decrease the quality of hawthorn berries from macro and micro perspectives., 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 Elsevier Ltd. All rights reserved.)

International trade in horticultural produce happens under phytosanitary inspection and production protocols. Fruit inspection typically involves the sampling and inspection of either 600-pieces or 2% of packed product within a single consignment destined for export, with the purpose of certification (typically with 95% confidence) that the true infestation level within the consignment in question doesn't exceed a pre-specified design prevalence. Sampling of multiple consignments from multiple production blocks in conjunction with pre-harvest monitoring for pests can be used to provide additional inference on the prevalence of infested fruit within an overall production system subject to similar protocols. Here we develop a hierarchical Bayesian model that combines in-field monitoring data with consignment sample inspection data to infer the prevalence of infested fruit in a production system. The results illustrate how infestation prevalence is influenced by the number of consignments inspected, the detection efficacy of consignment sampling, and in-field monitoring effort and sensitivity. Uncertainty in inspection performance, monitoring methods, and exposure of fruit to pests is accommodated using statistical priors within a Bayesian modelling framework. We demonstrate that pre-harvest surveillance with a sufficient density of traps and moderate detection sensitivity can provide 95% belief that the prevalence of infestation is below 1 × 10 - 6 . In the absence of pre-harvest monitoring, it is still possible to gain high confidence in a very low prevalence of infestation ( < 1 × 10 - 5 ) on the basis of multiple clean samples if the inspection sensitivity during consignment sampling is high and sufficient consignments are inspected. Our work illustrates the cumulative power of in-field surveillance and consignment sampling to update estimates of infestation prevalence., (© 2024. Crown.)

Invasive Anoplophora glabripennis recently became established in Japan and has caused heavy damage to several street-tree species. Overseas, A. glabripennis infests trees of the genera Acer and Populus as common host plants, and Malus, Pyrus, and Prunus (Rosaceae), including apple, pear, and plum trees; it therefore poses a potential risk to the production of economically valuable fruits in Japan. Fruit farms in areas already invaded by A. glabripennis are now threatened with tree infestation. We aimed to determine the potential damage to major fruit species in Japan. In the laboratory, we determined if the adult beetle is attracted to the odor of each of these tree species' branches; two confirmed host plant species and five Rosaceae fruit species, as well as its feeding preferences among branches of one host plant and the five fruit trees and its oviposition preferences among them. Among the fruit species, cherry branch had the highest rate of odor orientation by males. The feeding-preference assay showed that, besides the host plant, Japanese pear was the most consumed among the fruit trees. The potential risk of A. glabripennis laying eggs on fruit-tree branches was high for Japanese pear and above zero for plum, apple, and cherry branches., (© 2024. The Author(s).)

Apple, a vital fruit crop worldwide and a major crop grown in northern parts of India, acts as a backbone for the survival and livelihood of the farming community. However, it is prone to severe damage from insect pests and diseases. In the past few years, due to erratic weather patterns, there has been an alarmingly increased infestation of different insect pests, both invasive and non-invasive, resulting in substantial economic losses to this industry. One similar case was seen in the Himalayan region of India, where the apple crop in Karewas was heavily damaged by some invasive fruit borer, feeding on pulp and making galleries to reach the seed locules, thereby destroying the seed material. To ascertain the true identity of this pest, a study based on morpho-molecular characterization of this pest was carried out in the Faculty of Agriculture, Wadura, SKUAST Kashmir, India, during the years 2021 and 2022. The invasive fruit samples were collected from apple orchards at different locations (experimental sites) in North India by installing delta sticky traps @ 5 traps/ha for moth collection. The fruit-boring larvae and pupa were also collected and reared in confined chambers of the laboratory with controlled temperature and humidity. All the laboratory investigations were conducted at the Division of Entomology, FoA, Wadura, SKUAST Kashmir. During the investigation, it was confirmed that the invasive borer is a codling moth (Cydia pomonella L.), a member of the family Tortricidae, order Lepidoptera, which was earlier having quarantine importance in India, as it was only present in the Ladakh region of India. From the phylogenetic analysis of sample sequences, the species of codling moth present at all experimental sites was more identical to the codling moth sequence from Leh (the northernmost arid region), India. Further, the study of life cycle and voltinism revealed that codling moth completes three generations per year in Kashmir, with a single cycle lasting up to 2.5 months. However, the timing of various generations varies, depending on prevailing weather conditions at specific locations and times. The number of generations completed by codling moth in Kashmir conditions during apple growing season was three full and a half overwintering generations. The study of the life cycle of this important pest was necessary to know the weak link for its effective management and to prevent the economic loss in apple fruit., (© 2024. The Author(s).)

Background: Consuming contaminated raw vegetables and fruits is one of the primary means of parasite transmission to humans. Periodic monitoring of parasitic contamination in these food items is a crucial step in preventing the spread of parasitic disease in the community. This study was aimed at detecting intestinal parasitic contamination caused by consuming raw vegetables and fruits sold in three open-air markets and its associated factors in peri-urban areas of Jimma City, Oromia, Ethiopia., Methods: A cross-sectional study was conducted on fruits and vegetables collected from three peri-urban open-aired markets (namely; Hora Gibe, Bore and Jiren markets) in peri-urban areas of Jimma City between July and September 2021. A total of 187 fresh vegetable samples and 188 fruits were collected and examined for intestinal parasite contamination. About 200g of fruit and vegetable samples were processed and examined microscopically for parasite contamination, utilizing direct wet mount and modified Zeihl-Neelson staining methods in accordance with standard protocols. A structured questionnaire was used to collect data on the socio-demographic characteristics of vendors and risk factors for fruit and vegetable contamination. All data were analyzed using SPSS version 20.0., Result: Of the 187 fresh vegetable samples and 188 fresh vegetable samples, 105 (56.1%) and 68/188 (36.2%) of vegetables and fruit samples, respectively, were found contaminated with one or more intestinal parasites. Remarkably, high level of contamination in fresh vegetable samples was recorded both in Carrot (Daucus carota) 63.8% (30/46) and Lettuce (Lactuca sativa) 63.1% (29/46) while Green pepper (Capsicum spp.) is the least contaminated. In fruit samples, Avokado (Persea americana) 42.6% (20/47) and Banana (Musa acuminata) 14.9% (7/47) were the most and the least commonly contaminated items respectively. The identified helminthes and protozoans were Ascaris lumbricoides, Strongyloides stercoralis, Hymenolepis nana, Entamoeba histolytica/dispar, Giardia lamblia, Cryptosporidium spp., Toxocara spp. And Fasciola spp. The most predominant parasite encountered was A. lumbricoides 46(12.3%) whereas both Toxocara spp. 12(6.9) and Fasciola spp. 2(0.5) were the least detected parasites. It is worth-mentioned that the rate of contamination in Bore market (38.15%) was higher compared with Jiren market (34.7%) and Hora Gibe market (27%). However, the rate of contamination in vegetables and fruit obtained from the three district was non-significant (p = 0.19). Contamination was more common in vegetables than fruits (AOR = 5.78, p<0.001). It was also observed that decreased parasitic contamination was significantly associated with washing the products before displaying it for selling (p < 0.001)., Conclusion: The study has identified a high rate of raw vegetables and fruits contaminated with intestinal helminthes and protozoan. Contaminated fresh vegetables and fruits in open-aired peri-urban markets of Jimma city, Ethiopia may play a significant role in transmission of intestinal parasitic infections to humans, particularly A. lumbricoides infection. Therefore, it is urgently needed for health authorities to educate the public on the proper handling of vegetables and fruits prior to consumption., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Zeynudin et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The risk of foodborne parasite infection linked to the consumption of contaminated fresh produce has long been known. However, despite epidemiological links between the outbreaks and contaminated berries, few studies have assessed the magnitude of parasite contamination on fresh produce sold in Europe. The present study was aimed to address the knowledge gap on parasite contamination of berries sold in Norway. Samples of blueberries, strawberries, and raspberries were analysed by multiplex qPCR for detection of Echinococcus multilocularis, Toxoplasma gondii, and Cyclospora cayetanensis. In addition, a simplex qPCR method was employed for detecting contamination of the berries with Cryptosporidium spp. A total of 820 samples of berries, each of around 30 g (274 samples of blueberries, 276 samples of raspberries, and 270 samples of strawberries), were analysed. We found an overall occurrence of 2.9%, 6.6%, and 8.3% for T. gondii, C. cayetanensis, and Cryptosporidium spp., respectively, whereas E. multilocularis was not detected from any of the samples investigated. Strawberries and raspberries were most often contaminated with Cryptosporidium spp., whereas blueberries were contaminated mostly with C. cayetanensis. Detection of parasite contaminants on fresh berries indicates the need for a system to ensure the parasitological safety of fresh berries., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Bactrocera oleae , the olive fruit fly, is one of the most important pests affecting the olive fruit, causing serious quantitative and qualitative damage to olive oil production. In this study, the changes induced by B. oleae infestation in the biosynthesis of volatile and phenolic compounds in olive (cvs. Picual, Manzanilla, and Hojiblanca) have been analyzed. Despite cultivar differences, the oils obtained from infested fruits showed a significant increase in the content of certain volatile compounds such as ( E) -hex-2-enal, ethanol, ethyl acetate, and β-ocimene and a drastic decrease of the phenolic contents. The impact of those changes on the inferred quality of the oils has been studied. In parallel, the changes induced by the attack of the olive fly on the expression of some key genes in the biosynthesis of volatile and phenolic compounds, such as lipoxygenase, β-glucosidase, and polyphenol oxidase, have been analyzed. The strong induction of a new olive polyphenol oxidase gene ( OePPO2) explains the reduction of phenolic content in the oils obtained from infested fruits and suggest the existence of a PPO-mediated oxidative defense system in olives.

The coffee berry borer (CBB) Hypothenemus hampei Ferrari is the most serious pest of coffee worldwide. Management of the CBB is extremely difficult because its entire life cycle occurs inside the fruit, where it is well protected. Knowing which life stages contribute most to population growth, would shed light on the population dynamics of this pest and help to improve CBB management programs. Two staged-classified matrices were constructed for CBB populations reared in the lab on artificial diets and CBB populations from artificial infestations in the field. Matrices were used to determine demographic parameters, to conduct elasticity analyses, and to perform prospective perturbation analysis. Higher values of the intrinsic rate of natural increase (rm) and population growth rate (λ): were observed for CBB populations growing in the lab than in the field (rm: 0.058, λ: 1.74 lab; rm: 0.053, λ: 1.32 field). Sensitivity values for both CBB populations were highest for the transitions from larva to pupa (G2: 0.316 lab, 0.352 field), transition from pupa to juvenile (G3: 0.345 lab, 0.515 field) and survival of adult females (P5: 0.324 lab, 0.389 field); these three vital rates can be important targets for CBB management. Prospective perturbation analyses indicated that an effective management for the CBB should consider multiple developmental stages; perturbations of >90% for each transition are necessary to reduce λ to <1. However, when the three vital rates with highest sensitivity are impacted at the same time, the percentage of perturbation is reduced to 25% for each transition; with these reductions in survival of larvae, pupae and adult females the value of λ was reduced from 1.32 to 0.96. Management programs for CBB should be focused on the use of biological and cultural measures that are known to affect these three important targets., Competing Interests: The authors have declared that no competing interests exist.

Background: Diseases and pests have a profound effect on a yearly harvest and productivity in agriculture. A precise and accurate detection of the diseases and pests could facilitate timely treatment and management of the diseases and pests and lessen the resultant loss in economy and health. Herein, we propose an improved design of the disease detection system for plant images., Methods: Built upon the two-stage framework of object detection neural networks such as Mask R-CNN, the proposed network involves three types of extensions, including the addition of additional level of feature pyramids to improve the exploration and proposal of candidate regions, the aggregation of feature maps from all levels of feature pyramids per candidate region to fully exploit the information from feature pyramids, and the introduction of a squeeze-and-excitation block to the construction of feature pyramids and the aggregated feature maps to improve the representation of feature maps., Results: The proposed network was evaluated using 74 images of infected apple fruits. In 3-fold cross-validation, the proposed network achieved averaged precision (AP) of 72.26, AP at 0.5 threshold of 88.51 and AP at 0.75 threshold of 82.30. In the comparative experiments, the proposed network outperformed the other competing networks. The utility of the three extensions was also demonstrated in comparison to Mask R-CNN., Conclusions: The experimental results suggest that the proposed network could identify and localize the symptom of the disease with high accuracy, leading to an early diagnosis and treatment of the disease, and thus holding the potential for improving crop yield and quality., Competing Interests: The authors have declared that no competing interests exist.

Eriophyid mites are commonly found on the leaf surface of different plant species. In the present study, a novel virus associated with an eriophyid mite species was detected using high-throughput sequencing (HTS) of total RNA from fruit tree leaves, primarily growing under greenhouse conditions. The complete genome sequence was characterized using rapid amplification of cDNA ends followed by Sanger sequencing, revealing a genome of 8885 nucleotides in length. The single positive-stranded RNA genome was predicted to encode typical conserved domains of members of the genus Iflavirus in the family Iflaviridae. Phylogenetic analysis showed this virus to be closely related to the unclassified iflavirus tomato matilda associated virus (TMaV), with a maximum amino acid sequence identity of 59% in the RNA-dependent RNA polymerase domain. This low identity value justifies the recognition of the novel virus as a potential novel iflavirus. In addition to a lack of graft-transmissibility evidence, RT-PCR and HTS detection of this virus in the putative host plants were not consistent through different years and growing seasons, raising the possibility that rather than a plant virus, this was a virus infecting an organism associated with fruit tree leaves. Identification of Tetra pinnatifidae HTS-derived contigs in all fruit tree samples carrying the novel virus suggested this mite as the most likely host of the new virus (p-value < 1e-11), which is tentatively named "eriophyid mite-associated virus" (EMaV). This study highlights the importance of a careful biological study before assigning a new virus to a particular plant host when using metagenomics data., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

Originally from Asia, Drosophila suzukii Matsumura is a global pest of economically important soft-skinned fruits. Also commonly known as spotted wing drosophila, it is largely controlled through repeated applications of broad-spectrum insecticides by which resistance has been observed in the field. There is a pressing need for a better understanding of D. suzukii biology and for developing alternative environmentally friendly methods of control. The RNA-guided Cas9 nuclease has revolutionized functional genomics and is an integral component of several recently developed genetic strategies for population control of insects. Here, we describe genetically modified strains that encode three different terminators and four different promoters to express Cas9 robustly in both the soma and/or germline of D. suzukii . The Cas9 strains were rigorously evaluated through genetic crossing to transgenic strains that encode single-guide RNAs targeting the conserved X-linked yellow body and white eye genes. We find that several Cas9/gRNA strains display remarkably high editing capacity. Going forward, these tools will be instrumental for evaluating gene function in D. suzukii and may even provide tools useful for the development of new genetic strategies for control of this invasive species.

Background: Kiwifruit is an important horticultural crop all over the world and its development is important in Argentina. This dioecious crop has a short blooming period with nectarless flowers, and its fruit production depends on cross-pollination. Here, we tested whether kiwifruit quality increases by using honeybees exposed to female flowers treated with an artificial fragrance. The three experimental treatments were: A, sprinkled female flowers with 1:1 sugar syrup + Lavandula hybrida extract solution (a new attractant substance especially developed for this study named Lavandin Grosso); B, sprinkled female flowers with 1:1 water + sugar syrup (female flowers with additional sugar syrup reward); C (control; female flowers exposed to honeybees)., Results: The results showed a higher number of visits of honeybees to the female flowers sprinkled with the attractant substance, Lavandin Grosso, as well as higher fruit quality (weight, number of seeds, regularity in fruit size)., Conclusion: Our study demonstrates the potential of fragrance-treated flowers to improve yield production in kiwifruit. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

To investigate the presence of Cyclospora cayetanensis, Toxoplasma gondii and Echinococcus spp. in fresh produce sold in Italy, 324 locally produced 'ready-to-eat' (RTE) mixed-salad packages belonging to three brands and 324 berries packages (blueberries and blackberries imported from Peru and Mexico, respectively, and raspberries grown in Italy) were purchased at retail. Nine individual packages from each of the six types of fresh produce were collected monthly for one year, and with the same produce pooled, this resulted in a total of 72 pools for the whole year. Using microscopy (FLOTAC), a Cyclospora-like oocyst was detected in a blueberry sample and a taeniid egg was detected in a RTE-salad sample. Molecular tools confirmed these to be C. cayetanensis and Echinococcus multilocularis, respectively. Toxoplasma gondii was not detected in any of the samples. This study shows for the first time in Europe that imported berries on the Italian market may be contaminated with C. cayetanensis and RTE salads grown in Italy with E. multilocularis. The results indicate a new epidemiological scenario and highlight that current management of fresh produce, locally produced or imported, does not ensure products are free from parasite contamination., (Copyright © 2021. Published by Elsevier Ltd.)

Chemosensory proteins (CSPs) are important for insect chemoreception, which bind, solubilize and transport hydrophobic chemical molecules from external environment to dendrite membrane of chemosensory neurons. Moreover, CSPs are also involved in non-sensory physiological activities. The peach fruit borers Carposina sasakii Matsumura (Lepidoptera: Carposinidae) seriously damage fruit trees and their chemoreception mainly occurs in the adult stage. We identified 10 putative CSPs (CsasCSP1 ~ CsasCSP10) from head transcriptomes of C. sasakii adult males and females, all of which are classic CSPs that have 4 conserved cysteines with a spacing pattern C 1 -X 6 -C 2 -X 17 - 18 -C 3 -X 2 -C 4 . Their phylogenetic characteristics were also described. An analysis using fluorescence quantitative PCR showed CsasCSP2 has the highest level of expression in the heads, so it is more likely to be involved in C. sasakii chemoreception than the other C. sasakii CSPs. CsasCSP1, CsasCSP3, CsasCSP4, CsasCSP6, CsasCSP7 and CsasCSP8 are expressed dominantly in the wings; CsasCSP5 and CsasCSP10 have the highest expression level in the thoraxes; CsasCSP9 is dominantly and equally expressed in the thoraxes and abdomens. This study contributes to understanding physiological functions of C. sasakii CSPs and chemosensory mechanism at C. sasakii molecular level., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Although multiple outbreak clusters of Cyclospora cayetanensis have been traced back to consumption of dishes in Mexican-style restaurants, the FDA Bacteriological Analytical Manual (BAM) does not currently provide methods to detect C. cayetanensis in dishes that contain multiple produce ingredients, such as salsas and guacamole. These complex food matrices also may contain high levels of fats, which can interfere with the detection. Several modifications to the BAM Chapter 19b method (washing produce, DNA extraction, and a TaqMan real-time PCR assay targeting the 18S rRNA gene of C. cayetanensis) were assessed with the goal to detect as few as 5 oocysts of C. cayetanensis in 25 g samples of commercial salsa/pico de gallo, guacamole, and salsa verde. Both freshly prepared and frozen versions of these foods were seeded with 5, 10 and 200 oocysts. For salsa samples, using a gentler washing step than recommended by BAM, we achieved detection of 5 oocysts in the samples (81.8%, n = 11). Increasing the amount of Alconox® in the wash solution to 1%, rather than the 0.1% used in BAM, and adjusting the DNA extraction protocol to process large wash pellets, enabled detection of 5 oocysts in guacamole. To reach the desired level of detection in salsa verde, two types of modifications were necessary: gentler washing and DNA extraction modifications. The use of these same method modifications on previously frozen food samples, provided levels of detection similar to those achieved with fresh dishes. Our modifications enabled robust and reproducible detection of C. cayetanensis in multi-ingredient Mexican dishes, detecting as few as 5 oocysts in 25 g samples. Validating and deploying effective methods to detect C. cayetanensis in high risk fresh produce and prepared dishes are critically important for prevalence studies and outbreak investigations of this parasite., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Mangoes are tropical fruits appreciated worldwide but are extremely perishable, being susceptible to decay, pest infestation and fungal diseases. Using the flavorful and highly valued 'Manila' cultivar, we examined the effect of second-generation chitosan coatings on shelf-life, phenolic compound variation, phytohormones, pest infestation by fruit flies (Anastrepha obliqua) and anthracnose disease caused by the fungus Colletotrichum gloeosporioides., Results: We observed almost total elimination of A. obliqua eggs with 10 and 20 g L -1 chitosan in diluted acetic acid and a five- to sixfold reduction in anthracnose damage. Treatment with 20 g L -1 chitosan also extended the shelf-life. External (skin) and internal (pulp) discoloration processes were delayed. Fruit firmness was higher when compared with control and acetic acid treatments, and total soluble solids were lower in chitosan-treated fruit. Targeted and non-targeted metabolomics analyses on chitosan-coated fruit identified some phenolic compounds related to the tannin pathway. In addition, abscisic acid and jasmonic acid in the peel were downregulated in chitosan-coated mango peels. Both phytohormones and phenolic content may explain the reduced susceptibility of mangoes to anthracnose development and A. obliqua egg eclosion or larval development., Conclusions: We conclude that chitosan coatings represent an effective postharvest treatment that significantly reduces anthracnose disease, inhibits A. obliqua egg eclosion and significantly extends 'Manila' mango shelf-life, a key factor currently inhibiting large-scale commercialization of this valuable fruit. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Lower elevations are generally thought to contain a greater abundance and diversity of insect communities and their natural enemies than higher elevations. It is less clear, however, how changes in seasons influence this pattern. We conducted a 2-year study (2013‒2014) in guava orchards located in a tropical Andean forest of Peru to investigate differences in fruit flies (Diptera: Tephritidae) and their parasitoid communities at two elevations and over two seasons. Fruit fly traps were installed, monitored, and guava fruits were sampled from eight orchards at low (800-950 m above sea level) and high (1,700-1,900 m above sea level) elevations and during the dry and rainy seasons. At each orchard, adult fruit fly trap captures and emergence of fruit flies and their parasitoids from guava fruit were quantified to determine their abundance and species composition. There was a greater abundance and species richness of fruit flies captured in traps at lower elevations, as well as higher abundance and species evenness of fruit flies that emerged from fruit, indicating that lower elevations are associated with larger fruit fly populations. The abundance, species richness and diversity of parasitoids were also greater at lower elevations. Consequently, guava fruit infestation and fruit fly parasitism rates were also greater at lower elevations. Seasonality also influenced fruit fly populations with a greater number of flies emerging from guava fruit and more fruit infested in the rainy season. However, seasonality had no effect on parasitoid population parameters or rate of parasitism, nor did it interact with elevation as an influence of populations of fruit flies or their parasitoids in guava orchards. This study highlights the importance of examining both elevation and seasonality for a better understanding of the population dynamics of fruit flies and their parasitoids in tropical agroecosystems., Competing Interests: The authors have declared that no competing interests exist.

Drosophila suzukii (spotted wing drosophila, SWD) is a pandemic quarantine pest that attacks mostly red fruits. The high number of life cycles per year, its ability to rapidly invade and spread across new habitats, and highly polyphagous nature, makes this a particularly aggressive invasive species, for which efficient control methods are currently lacking. The use of native natural predators is particularly promising to anchor sustainable and efficient measures to control SWD. While several field studies have suggested the presence of potential predatory species in infested orchards, only a few confirmed the presence of SWD DNA in predators' gut content. Here, we use a DNA-based approach to identify SWD predators among the arthropod diversity in South Europe, by examining the gut content of potential predator specimens collected in SWD-infested berry fields in North Portugal. These specimens were morphologically identified to the family/order, and their gut content was screened for the presence of SWD DNA using PCR. New SWD predatory taxonomical groups were identified, as Opiliones and Hemerobiidae, in addition to known SWD predators, such as Hemerobiidae, Chrysopidae, Miridae, Carabidae, Formicidae and Araneae. Additionally, the presence of a spider family, Uloboridae, in the orchards was recorded for the first time, posing this family as another SWD-candidate predator. This study sets important bases to further investigate the potential large-scale use of some of these confirmed predator taxa for SWD control in South Europe., Competing Interests: The authors have declared that no competing interests exist.

Biosecurity interception records are crucial data underlying efforts to predict and manage pest and pathogen introductions. Here we present a dataset containing information on imported plant products inspected by the South African Department of Agriculture's laboratories between 1994 and 2019 and the contaminant organisms found on them. Samples were received from border inspectors as either propagation material (e.g. plants) or material for immediate use (e.g. fruit). Material for immediate use was further divided into two sample categories, depending on if contaminants were seen/suspected by the border official or not: intervention or audit samples. The final dataset consists of 25,279 records, of which 30% tested positive (i.e. had at least one contaminant) and 13% had multiple contaminants. Of the 13,731 recorded contaminants, fungi (41%), mites (37%) and insects (19%) were most common. This dataset provides insight into the suite of taxa transported along the plant import pathway and provides an important resource for analyses of contaminant organisms in international trade, which can inform strategies for risk assessment, pathway management and biosecurity protocols.

The coffee berry borer (CBB, Hypothenemus hampei) is the most serious insect pest of coffee worldwide; understanding the dynamics of its reproduction is essential for pest management. The female CBB penetrates the coffee berry, eats the seed, and reproduces inside it. A mathematical model of the infestation progress of the coffee berry by the CBB during several coffee seasons is formulated. The model represents the interaction among five populations: uninfested, slightly infested, and severely infested coffee berries, and free and encapsulated CBBs. Coffee harvesting is also included in the model. A one-dimensional map is derived for tracking the population dynamics subject to certain coffee harvesting percentages over several seasons. Stability analysis of the map's fixed points shows that CBB infestation could be eliminated or controlled to a specific level over multiple seasons of coffee harvesting. However, the percent of coffee harvesting required is determined by the level of CBB infestation at the beginning of the first season and in some cases it is impossible to achieve that percentage., 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 © 2021 Elsevier Inc. All rights reserved.)

Cyclospora cayetanensis is a parasite causing cyclosporiasis (an illness in humans). Produce (fruits, vegetables, herbs), water and soil contaminated with C. cayetanensis have been implicated in human infection. The objective was to conduct a scoping review of primary research in English on the detection, epidemiology and control of C. cayetanensis with an emphasis on produce, water and soil. MEDLINE® (Web of ScienceTM), Agricola (ProQuest), CABI Global Health, and Food Science and Technology Abstracts (EBSCOhost) were searched from 1979 to February 2020. Of the 349 relevant primary research studies identified, there were 75 detection-method studies, 40 molecular characterisation studies, 38 studies of Cyclospora in the environment (33 prevalence studies, 10 studies of factors associated with environmental contamination), 246 human infection studies (212 prevalence/incidence studies, 32 outbreak studies, 60 studies of environmental factors associated with non-outbreak human infection) and eight control studies. There appears to be sufficient literature for a systematic review of prevalence and factors associated with human infection with C. cayetanensis. There is a dearth of publicly available detection-method studies in soil (n = 0) and water (n = 2), prevalence studies on soil (n = 1) and studies of the control of Cyclospora (particularly on produce prior to retail (n = 0)).

Host plant attributes are essential factors determining the population dynamics of herbivorous insects. The developmental stage of host plants, in particular, may affect the biology of Grapholita molesta (Busck), a possibility that has rarely been examined. Here we assessed the effect of developmental stage of plum, peach, and apple fruits on the development and fecundity performance of G. molesta, along with an examination of the firmness and sugar content of the fruits. Among the fruits collected earliest (May 31), plum and apple were better food sources for G. molesta compared to peach in terms of development, reproduction, and life table parameters. However, despite the higher sugar content in peach, G. molesta larvae showed a lower rate of fruit penetration in peach, probably due to fruit firmness. In the later-collected fruit (June 25), both peach and apple were better than plum, as peach and apple were softer and had higher sugar content. Nevertheless, the penetration rate of larva was still low in peach probably due to pubescence on the fruit surface. Although the plum fruits in the later collection date were softer with higher sugar content, there was a negative impact on the development and reproduction because fruits started to liquefy earlier. In conclusion, the developmental stage of fruits with changes in fruit firmness or sugar content affected the development and reproduction of G. molesta, and apple would be the best food source.

Controlling the cosmopolitan pest Drosophila suzukii (spotted wing drosophila) is a challenge for fruit growers. A promising agent for biological control of that pest are parasitoid wasps. Especially the widespread pupal parasitoid Trichopria drosophilae had shown the ability to parasitise the pest fly. However, as a biocontrol agent, parasitoids can only be effective when they prefer the pest to other insects. Until now studies have been inconsistent concerning the preference of T. drosophilae for D. suzukii and whether the preference depends on pupal volume. To clarify this inconsistency, we used video recordings of parasitisation experiments with a set up to observe the direct host preference of the parasitoid. Additionally, the volume of each host pupa was measured. We found significant preference of T. drosophilae for D. suzukii pupae independent of the pupal size and of the host species the wasps were reared on. The article also discusses the sex ratio and the success of the parasitoid in the different pupae characteristics.

Drosophila suzukii flies cause economic losses to fruit crops globally. Previous work shows various Drosophila species are attracted to volatile metabolites produced by individual fruit associated yeast isolates, but fruits naturally harbour a rich diversity of yeast species. Here, we report the relative attractiveness of D. suzukii to yeasts presented individually or in combinations using laboratory preference tests and field trapping data. Laboratory trials revealed four of 12 single yeast isolates were attractive to D. suzukii, of which Metschnikowia pulcherrima and Hanseniaspora uvarum were also attractive in field trials. Four out of 10 yeast combinations involving Candida zemplinina, Pichia pijperi, M. pulcherrima and H. uvarum were attractive in the laboratory. Whilst a combination of M. pulcherrima + H. uvarum trapped the greatest number of D. suzukii in the field, the efficacy of the M. pulcherrima + H. uvarum combination to trap D. suzukii was not significantly greater than traps primed with volatiles from only H. uvarum. While volatiles from isolates of M. pulcherrima and H. uvarum show promise as baits for D. suzukii, further research is needed to ascertain how and why flies are attracted to certain baits to optimise control efficacy.

&lt;b&gt;Background and Objective:&lt;/b&gt; Control of the stored date insects by an eco-friendly method is essential to maintain the fruit for prolonged periods, especially that dates are eaten fresh. The search for a safe method, as an alternative to commonly used chemical pesticides, is needed. The objective is to investigate CO&lt;sub&gt;2&lt;/sub&gt; enriched atmosphere on the Mortality Percentage (MP) of the Saw-toothed grain beetle, &lt;i&gt;O. surinamensis&lt;/i&gt;, life stages with special emphasis on reducing damage to stored dates. &lt;b&gt;Materials and Methods:&lt;/b&gt; The effect of high levels of CO&lt;sub&gt;2&lt;/sub&gt; as an alternative control method against the Saw-toothed grain beetle, &lt;i&gt;Oryzaephilus surinamensis&lt;/i&gt; L., at different life stages was studied on infested 'Saqie' dates. Four CO&lt;sub&gt;2&lt;/sub&gt; pressures (25, 50, 75 and 90 kPa, balance is nitrogen) were tested for 6, 12, 18, 24, 48, 72 and 96 hrs intervals. The response of different life stages of &lt;i&gt;O. surinamensis&lt;/i&gt; to the different treatments varied according to CO&lt;sub&gt;2&lt;/sub&gt; level, developmental stage and exposure period. &lt;b&gt;Results:&lt;/b&gt; Mortality (%) was higher during the larval stage, followed by adults, pupae and eggs, in descending order. The larvae and adult stages were more sensitive to CO&lt;sub&gt;2&lt;/sub&gt; treatment than the pupal and egg. Exposure time was more effective on eggs, larval and adult MP than the CO&lt;sub&gt;2&lt;/sub&gt; atmosphere level. Mortality% at 96 hrs exposure time was almost 100% with CO&lt;sub&gt;2&lt;/sub&gt; atmospheres of 50, 75 and 90 kPa. Mathematical equations were developed to model the relationship between mortality% and CO&lt;sub&gt;2&lt;/sub&gt; treatments using multiple regression analyses for each life stage. &lt;b&gt;Conclusion:&lt;/b&gt; The results confirmed that CO&lt;sub&gt;2&lt;/sub&gt; could be applied to final food products during packaging to control the residual occurrence of insect pests after storage and before the packaging process to prevent further infestation in the final packages.

Finding a suitable oviposition site is a challenging task for a gravid female fly, because the hatched maggots have limited mobility, making it difficult to find an alternative host. The oriental fruit fly, Bactrocera dorsalis, oviposits on many types of fruits. Maggots hatching in a fruit that is already occupied by conspecific worms will face food competition. Here, we showed that maggot-occupied fruits deter B. dorsalis oviposition and that this deterrence is based on the increased β-caryophyllene concentration in fruits. Using a combination of bacterial identification, volatile content quantification, and behavioral analyses, we demonstrated that the egg-surface bacteria of B. dorsalis, including Providencia sp. and Klebsiella sp., are responsible for this increase in the β-caryophyllene contents of host fruits. Our research shows a type of tritrophic interaction between micro-organisms, insects, and insect hosts, which will provide considerable insight into the evolution of insect behavioral responses to volatile compounds., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Body size is commonly associated with biological features such as reproductive capacity, competition, and resource acquisition. Many studies have tried to understand how these isolated factors can affect the body pattern of individuals. However, little is known about how interactions among species in multitrophic communities determine the body shape of individuals exploiting the same resource. Here, we evaluate the effect of fruit infestation, parasitism rate, and seed biomass on size, allometric and asymmetric patterns of morphological structures of insects that exploit the same resource. To test it, we measured 750 individuals associated with the plant Senegalia tenuifolia (Fabaceae), previously collected over three consecutive years. Negative allometry was maintained for all species, suggesting that with increasing body size the body structure did not grow proportionally. Despite this, some variations in allometric slopes suggest that interactions in a multitrophic food web can shape the development of these species. Also, we observed a higher confidence interval at higher categories of infestation and parasitism rate, suggesting a great variability in the allometric scaling. We did not observe fluctuating asymmetry for any category or species, but we found some changes in morphological structures, depending on the variables tested. These findings show that both allometry and morphological trait measurements are the most indicated in studies focused on interactions and morphometry. Finally, we show that, except for the fluctuating asymmetry, each species and morphological structure respond differently to interactions, even if the individuals play the same functional role within the food web., Competing Interests: The authors have declared that no competing interests exist.

Organic waste is a rapidly increasing problem due to the growth of the agricultural production needed to meet global food demands. Development of sustainable waste management solutions is essential. Black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae) (BSF), larvae are voracious consumers of a wide range of organic materials ranging from fruits and vegetables to animal remains, and manure. Thanks to this ability and considering the larval high protein and lipid content, BSF larvae are a useful additive in animal feeds and biodiesel production. Unfortunately, the feasibility of using the black soldier fly as a tool for waste valorization and feed production has primarily been investigated at the benchtop scale. Thus, mobilization of current practices to an industrial scale is challenging because scaling up from small laboratory studies to large industrial studies is not necessarily linear. The goal of this study was to demonstrate the ability of the BSF to recycle organic waste at an industrial scale. To accomplish this goal, three organic waste streams were used (e.g., apples, bananas, and spent grain from a brewery) to test six diet treatments (1) apple, (2) banana, (3) spent grain, (4) apple and banana, (5) apple and spent grain, and (6) banana and spent grain. Working at scale of 10,000 BSF larvae life history traits, waste valorization, protein and lipid profiles were measured for each diet treatment. Differences were recorded across all variables, except substrate conversion, for larvae fed on fruit and spent grain (alone or with fruit). Growth rate significantly differed across treatments; larvae reared on spent grain grew twice as fast as those fed apples alone, but those reared on the apple and spent grain mixture produced twice as much insect biomass. However, it should be noted that larvae resulting from the apple diet contained 50% more fat than larvae fed the fruit and spent grain mixtures. Commonly-available organic wastes were successfully used at an industrial scale to produce BSF larvae that have the potential to substitute other sources of protein and lipids in different industrial applications. Industrialization efforts are encouraged to assess these impacts when integrating diverse ingredients into larval diets as a means to more precisely predict output, such as larval development time and final larval biomass.

Uncertainty about the taxonomic status and the specificity of a species commonly prevent its consideration as a candidate for biological control of pest organisms. Here we use a combination of molecular analysis and crossing experiments to gather evidence that the parasitoid wasp Ganaspis brasiliensis, a candidate for biological control of the invasive spotted wing drosophila Drosophila suzukii, is a complex of at least two cryptic species. Complementary experiments demonstrate that individuals from one genetic group readily parasitize several drosophila species regardless of their food source while individuals from the other one are almost exclusively specific to larvae feeding in ripening fruits. Because only D. suzukii attacks ripening fruits in its area of invasion, parasitoids from this second group appear to be well suited as a biological control agent. Our study demonstrates the need for a combination of biosystematics with biological and ecological investigations for the development of safe and efficient biological control programs.

Plant diseases bear names such as leaf blights, root rots, sheath blights, tuber scabs, and stem cankers, indicating that symptoms occur preferentially on specific parts of host plants. Accordingly, many plant pathogens are specialized to infect and cause disease in specific tissues and organs. Conversely, others are able to infect a range of tissues, albeit often disease symptoms fluctuate in different organs infected by the same pathogen. The structural specificity of a pathogen defines the degree to which it is reliant on a given tissue, organ, or host developmental stage. It is influenced by both the microbe and the host but the processes shaping it are not well established. Here we review the current status on structural specificity of plant-filamentous pathogen interactions and highlight important research questions. Notably, this review addresses how constitutive defence and induced immunity as well as virulence processes vary across plant organs, tissues, and even cells. A better understanding of the mechanisms underlying structural specificity will aid targeted approaches for plant health, for instance by considering the variation in the nature and the amplitude of defence responses across distinct plant organs and tissues when performing selective breeding., (© 2020 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.)

Rapid human population growth and associated urbanization lead to increased artificial illumination of the environment. By changing the natural light-dark cycle, artificial lighting can affect the functioning of natural ecosystems. Many plants rely on insects in order to reproduce but these insects are known to be disturbed by artificial light. Therefore, plant-insect interactions may be affected when exposed to artificial illumination. These effects can potentially be reduced by using different light spectra than white light. We studied the effect of artificial lighting on plant-insect interactions in the Silene latifolia-Hadena bicruris system using a field set-up with four different light treatments: red, green, white and a dark control. We compared the proportion of fertilized flowers and fertilized ovules as well as the infestation of fruits by Hadena bicruris, a pollinating seed predator. We found no difference in the proportion of fertilized flowers among the treatments. The proportion of fruits infested by H. bicruris was however significantly higher under green and white light and a significantly lower proportion of fertilized ovules was found under green light. We show that artificial light with different colours impacts plant-insect interactions differently, with direct consequences for plant fitness.

Because plant phenotypes can change in response to attacks by herbivores in highly variable ways, the distribution of herbivores depends on the occurrence of other herbivore species on the same plant. We carried out a field study to evaluate the co-occurrence of three coconut pests, the mites Aceria guerreronis (Acari: Eriophyidae), Steneotarsonemus concavuscutum (Acari: Tarsonemidae) and the moth Atheloca bondari (Lepidoptera: Pyralidae). The eriophyid mite Ac. guerreronis is the most important coconut pest around the world, whereas S. concavuscutum and At. bondari are economically important only in some areas along the Brazilian coast. A previous study suggested that the necrosis caused by Ac. guerreronis facilitates the infestation of At. bondari larvae. Because all three species infest the area under the perianths on coconuts and S. concavuscutum also causes necrosis that could facilitate At. bondari, we evaluated the co-occurrence of all three species. We found that the occurrence of At. bondari was positively associated with Ac. guerreronis, but negatively associated with S. concavuscutum. In addition, the two mite species showed negative co-occurrence. Atheloca bondari was found on nuts of all ages, but more on nuts that had fallen than on those on the trees, suggesting that nuts infested by At. bondari tend to fall more frequently. We discuss the status of At. bondari as a pest and discuss experiments to test the causes of these co-occurrence patterns.

Wild tomato species represent a rich gene pool for numerous desirable traits lost during domestication. Here, we exploited an introgression population representing wild desert-adapted species and a domesticated cultivar to establish the genetic basis of gene expression and chemical variation accompanying the transfer of wild-species-associated fruit traits. Transcriptome and metabolome analysis of 580 lines coupled to pathogen sensitivity assays resulted in the identification of genomic loci associated with levels of hundreds of transcripts and metabolites. These associations occurred in hotspots representing coordinated perturbation of metabolic pathways and ripening-related processes. Here, we identify components of the Solanum alkaloid pathway, as well as genes and metabolites involved in pathogen defense and linking fungal resistance with changes in the fruit ripening regulatory network. Our results outline a framework for understanding metabolism and pathogen resistance during tomato fruit ripening and provide insights into key fruit quality traits.

Spotted wing drosophila, Drosophila suzukii, is a serious invasive pest impacting the production of multiple fruit crops, including soft and stone fruits such as strawberries, raspberries and cherries. Effective control is challenging and reliant on integrated pest management which includes the use of an ever decreasing number of approved insecticides. New means to reduce the impact of this pest that can be integrated into control strategies are urgently required. In many production regions, including the UK, soft fruit are typically grown inside tunnels clad with polyethylene based materials. These can be modified to filter specific wavebands of light. We investigated whether targeted spectral modifications to cladding materials that disrupt insect vision could reduce the incidence of D. suzukii. We present a novel approach that starts from a neuroscientific investigation of insect sensory systems and ends with infield testing of new cladding materials inspired by the biological data. We show D. suzukii are predominantly sensitive to wavelengths below 405 nm (ultraviolet) and above 565 nm (orange & red) and that targeted blocking of lower wavebands (up to 430 nm) using light restricting materials reduces pest populations up to 73% in field trials.

Spotted-wing drosophila, Drosophila suzukii, and the anthracnose pathogen Colletotrichum fioriniae are an important insect pest and fungal disease of highbush blueberries, respectively, in the United States. However, whether C. fioriniae infection affects D. suzukii preference and performance remains unknown. Here, we conducted choice and no-choice studies to determine the repellent, oviposition-deterrent, and insecticidal effects of C. fioriniae on D. suzukii. In choice tests, blueberry fruit treated with anthracnose solutions containing spores from either field-collected infected fruit ('fruit') or a laboratory C. fioriniae culture ('colony') were less attractive to sexually mature D. suzukii females, but not males, than untreated fruit. The plant tissue (fruit or leaves) did not influence C. fioriniae repellency effects on D. suzukii. In no-choice tests, 55% fewer numbers of eggs were laid on, and 65% fewer adults emerged from, blueberry fruit treated with either the 'fruit' or 'colony' anthracnose solution than untreated fruit. Egg-to-adult D. suzukii survival was also 12% lower on C. fioriniae-infected fruit. No repellency or negative effects on survival were observed when C. fioriniae spores were filtered out of the solution. These findings will help efforts towards the discovery of microbial-derived repellent/oviposition-deterrent compounds that could be used in behavior-based management strategies for D. suzukii.

The coffee berry borer (CBB) is one of the main coffee pests in the world including Colombia. This pest is difficult to manage because of its cryptic habits and the continuous availability of coffee fruits. Among the new management strategies being tested is the use of volatile compounds as insect repellents. In this work, the behavioral response of female adult CBBs to terpenes previously identified in the CBB-repellent plant species Lantana camara was evaluated. α-Terpinene, (R)-limonene, farnesene and β-caryophyllene terpenes were tested via a Y-tube olfactometer in which ripe coffee fruits were accompanied by terpenes at concentrations between 25 and 200 ppm. Only β-caryophyllene induced a significant and consistent CBB repellent effect at all tested doses. The protective effect of microencapsulated β-caryophyllene was then determined under laboratory conditions by incorporating the terpene in a colloidosome-gel system at 2.8 × 10 5 ng/h in the middle of coffee fruits with adult CBBs. The coffee fruits in turn presented a decrease in fruit infestation. Furthermore, the protection of coffee fruits when β-caryophyllene gels were hung in coffee trees was evaluated in the field; infestations were artificially induced by the use of raisins (CBB-infested old coffee fruits) placed on the ground. Compared with unprotected trees, the trees treated with caryophyllene gels exhibited a 33 to 45% lower degree of infestation. Taken together, the results show that β-caryophyllene is a promising compound for an integrated pest management (IPM) program in commercial coffee plantations.

The potential public health impact of foodborne parasites (FBP) transmitted via contaminated fresh produces indicates the necessity for robust and reliable laboratory methods for their detection and identification on this infection vehicle. Standardization of methods for detection of common FBP in fresh produce is to be expected and ensuring that the DNA extraction approach is most appropriate for the FBP of interest and for the matrix being analyzed is also important. Therefore, the aim of the present study was to compare the efficacy of two commercially available DNA extraction procedures, the UNEX-based method and DNeasy PowerSoil kit in the detection of three protozoan parasites, C. cayetanensis, C. parvum, and T. gondii, on contaminated berries. Oocysts of each parasite were spiked into the pellets of raspberry and blueberry washes. The spiked pellets were then randomly assigned to DNA extraction using either the PowerSoil or UNEX method, with DNA extraction with both methods performed by two independent analysts. The detection rate when berry washes were spiked with 20 oocysts of C. cayetanensis, T. gondii, and C. parvum was 95%, 85%, and 40%, respectively, when using the PowerSoil kit; whereas the equivalent results using the UNEX method were 55%, 60%, and 5%, respectively. In addition, significantly lower C q values were achieved for each parasite in the samples spiked with 500 oocysts when the PowerSoil kit was used. Possible reasons for these results are discussed, and include the composition of both the beads and the buffers in each method., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Over the past decade, the spotted wing Drosophila, Drosophila suzukii, has invaded Europe and America and has become a major agricultural pest in these areas, thereby prompting intense research activities to better understand its biology. Two draft genome assemblies already exist for this species but contain pervasive assembly errors and are highly fragmented, which limits their values. Our purpose here was to improve the assembly of the D. suzukii genome and to annotate it in a way that facilitates comparisons with D. melanogaster. For this, we generated PacBio long-read sequencing data and assembled a novel, high-quality D. suzukii genome assembly. It is one of the largest Drosophila genomes, notably because of the expansion of its repeatome. We found that despite 16 rounds of full-sib crossings the D. suzukii strain that we sequenced has maintained high levels of polymorphism in some regions of its genome. As a consequence, the quality of the assembly of these regions was reduced. We explored possible origins of this high residual diversity, including the presence of structural variants and a possible heterogeneous admixture pattern of North American and Asian ancestry. Overall, our assembly and annotation constitute a high-quality genomic resource that can be used for both high-throughput sequencing approaches, as well as manipulative genetic technologies to study D. suzukii.

Spotted Wing Drosophila (Drosophila suzukii; Matsumura) is an invasive fruit fly with the ability to oviposit in a broad range of agriculturally valuable fruits. Volatile organic compounds (VOCs) produced by botanical oils may reduce D. suzukii's attraction to hosts and decrease survival, but it is unknown whether their efficacy varies across D. suzukii life stages or affects the survival and success of higher trophic levels. Through a series of laboratory bioassays, we evaluated the effects of peppermint (Mentha arvensis L.) oil produced VOCs on D. suzukii survival and the survival of and parasitism rates by a pupal parasitoid wasp, Pachycrepoideus vindemmiae (Rondani). First, we determined whether fumigation with peppermint oil VOCs at the pupal stage reduced adult emergence, and whether this depended on environmental conditions (i.e. soil moisture). Second, we evaluated whether fumigation with peppermint oil VOCs reduced or enhanced parasitism by the pupal parasitoid and whether this depended on the timing of peppermint oil VOC exposure (i.e. before, during, or after parasitoid access). Fumigation with VOCs of 4.5 mg of peppermint oil reduced D. suzukii emergence under moist soil conditions but dry soil had a similar effect on reducing adult emergence as peppermint oil presence. Peppermint oil VOC fumigation was toxic to adult P. vindemmiae, but developing P. vindemmiae were unaffected by peppermint oil VOC fumigation. Using peppermint essential oil as a fumigant may reduce D. suzukii emergence from the pupal stage. However, this could negatively impact P. vindemmiae dependent on the timing of application.

The pod wall of legumes is known to protect the developing seeds from pests and pathogens. However, the mechanism of conferring defense against insects has not yet been deciphered. Here, we have utilized 2-dimensional gel electrophoresis (2D-GE) coupled with mass spectrometry (MS/MS) to identify over expressed proteins in the pod wall of two different cultivars (commercial cultivar: JG 11 and tolerant cultivar: ICC 506-EB) of chickpea after 12 h of application of Helicoverpa armigera oral secretions (simulated herbivory). The assays were performed with a view that larvae are a voracious feeder and cause substantial damage to the pod within 12 h. A total of 600 reproducible protein spots were detected on gels, and the comparative analysis helped identify 35 (12 up-regulated, 23 down-regulated) and 20 (10 up-regulated, 10 down-regulated) differentially expressed proteins in JG 11 and ICC 506-EB, respectively. Functional classification of protein spots of each cultivar after MS/MS indicated that the differentially expressed proteins were associated with various metabolic activities. Also, stress-related proteins such as mannitol dehydrogenase (MADH), disease resistance-like protein-CSA1, serine/threonine kinase (D6PKL2), endoglucanase-19 etc. were up-regulated due to simulated herbivory. The proteins identified with a possible role in defense were further analyzed using the STRING database to advance our knowledge on their interacting partners. It decoded the involvement of several reactive oxygen species (ROS) scavengers and other proteins involved in cell wall reinforcement. The biochemical analysis also confirmed the active role of ROS scavengers during simulated herbivory. Thus, our study provides valuable new insights on chickpea-H.armigera interactions at the protein level.

Sulfoxaflor, the first commercially available sulfoximine insecticide, has been used for the control of sap-feeding insect pests such as plant bugs and aphids on a variety of crops. However, its sublethal effects on the mirid bug Apolygus lucorum, one of the key insect pests of Bt cotton and fruit trees in China, have not been fully examined. Here, we evaluated the demography and feeding behaviour of A. lucorum exposed to sulfoxaflor. The leaf-dipping bioassay showed that the LC10 and LC30 of sulfoxaflor against 3rd-instar nymphs of this insect were 1.23 and 8.37 mg L-1, respectively. The LC10 significantly extended the nymphal duration and decreased the oviposition period by 5.29 days and female fecundity by 56.99% in the parent generation (F0). The longer duration of egg, 5th-instar nymphs, preadult, and male adult longevity were observed in the F1 generation (F1) at LC10. At the LC30, the duration of egg and 1st-instar nymph, female adult longevity, and oviposition period of the F1 were significantly shorter, while the nymphal duration in the F0 and duration of 5th-instar nymphs, preadult survival rate, and male adult longevity in the F1 significantly increased. The net reproductive rate (R0), intrinsic rate of increase (r), and finite rate of increase (λ) in the F1 were not significantly affected by these two concentrations, whereas the mean generation time (T) was lower at the LC30. Additionally, the probe counts and cells mixture feeding time were markedly lengthened by the LC10 and LC30, respectively, when A. lucorum nymphs exposed to sulfoxaflor fed on Bt cotton plants without insecticides. These results clearly indicate that sulfoxaflor causes sublethal effects on A. lucorum and the transgenerational effects depend on the tested concentrations., Competing Interests: The authors have declared that no competing interests exist.

Ochratoxin A (OTA) is the most toxic member of ochratoxins, a group of toxic secondary metabolites produced by fungi. The most relevant species involved in OTA production in grapes is Aspergillus carbonarius . Berry infection by A. carbonarius is enhanced by damage to the skin caused by abiotic and biotic factors. Insect pests play a major role in European vineyards, and Lepidopteran species such as the European grapevine moth Lobesia botrana are undoubtedly crucial. New scenarios are also emerging due to the introduction and spread of allochthonous pests as well as climate change. Such pests may be involved in the dissemination of OTA producing fungi even if confirmation is still lacking and further studies are needed. An OTA predicting model is available, but it should be integrated with models aimed at forecasting L. botrana phenology and demography in order to improve model reliability.

Outbreaks and sporadic cases of Cyclospora cayetanensis have been linked to consumption of berries. The efficacy of the U.S. Food and Drug Administration (FDA) method for detection of C. cayetanensis was evaluated in fresh berries (blackberries, strawberries, blueberries and mixed berries) and in frozen mixed berries. The protocol included seeding with C. cayetanensis oocysts, produce washing, DNA extraction and a dual TaqMan assay. As few as five oocysts were detected in every type of fresh berry analyzed. All berry samples seeded with 200 oocysts were positive and all unseeded berry samples were negative. No significant differences were observed among any of the berry types analyzed in detection rates, C T values and estimated oocyst recovery percentages. Mixed berries were seeded and frozen for up to seven weeks. As few as five oocysts were also detected. No significant differences were observed in C. cayetanensis C T values between fresh and frozen mixed berries at any seeding level. In conclusion, the FDA BAM Chapter 19B method for the detection of Cyclospora was robust, consistent, and showed high sensitivity in all types of berries analyzed. Evaluation of the FDA detection method in berries will provide reliable laboratory support for surveillance programs and for outbreak investigations., (Published by Elsevier Ltd.)

The red-necked longhorn beetle (RLB) Aromia bungii (Fald.) is an emerging pest of stone fruit trees, native to East Asia, accidentally introduced in Europe (Germany and Italy) and Japan. Threatening seriously the stone fruit crops in Europe, RLB was added to both the EPPO A1 and priority pest lists of quarantine species. Molecular analyses highlighted that all specimens recovered in southern Italy share the same haplotype, different from the German one, supporting that the invasive process in Europe started from at least two independent introductions. To fill the existing gap of biological knowledge about A. bungii, several laboratory tests were carried out on specimens collected in the outbreak area of Naples (Italy). Results suggest a high biotic potential of the RLB Italian population. Females showed a short pre-oviposition period while the period of oviposition lasted about three weeks, with a rate of 24.2 eggs/day. Each female laid an average of 587.5 eggs and spawned the largest amount of eggs during the first week after emergence. Fed males live up to 62 days at 20 °C while fed females about 63 days at 25 °C. These results are crucial to draw up a multi-facet IPM approach against A. bungii in the outbreak areas.

Spotted-wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), is an invasive economic pest of soft-skinned and stone fruit across the globe. Our study establishes both a predictive generalized linear mixed model (GLMM), and a generalized additive mixed model (GAMM) of the dynamic seasonal phenology of D. suzukii based on four years of adult monitoring trap data in Wisconsin tart cherry orchards collected throughout the growing season. The models incorporate year, field site, relative humidity, and degree days (DD); and relate these factors to trap catch. The GLMM estimated a coefficient of 2.21 for DD/1000, meaning for every increment of 1000 DD, trap catch increases by roughly 9 flies. The GAMM generated a curve based on a cubic regression smoothing function of DD which approximates critical DD points of first adult D. suzukii detection at 1276 DD, above average field populations beginning at 2019 DD, and peak activity at 3180 DD. By incorporating four years of comprehensive seasonal phenology data from the same locations, we introduce robust models capable of using DD to forecast changing adult D. suzukii populations in the field leading to the application of more timely and effective management strategies., Competing Interests: The authors have declared that no competing interests exist.

Background: Toxoplasma gondii is the third most important contributor to health burden caused by food-borne illness. Ingestion of tissue cysts from undercooked meat is an important source of horizontal transmission to humans. However, there is an increasing awareness of the consumption of fresh fruit and vegetables, as a possible source for oocyst transmission, since this stage of the parasite can persist and remain infective in soil and water for long time. Herein, we outline findings related with detection of T. gondii oocysts in vegetables and berry fruits, which are usually raw consumed. The procedure includes the estimation of the number of oocysts., Methods: Food samples were collected from local producers and supermarket suppliers. Toxoplasma gondii oocysts were concentrated after washing the samples by applying high resolution water filtration and immunomagnetic separation (method 1623.1: EPA 816-R-12-001-Jan 2012), in order to (i) remove potential Cryptosporidium spp. oocysts and Giardia spp. cysts present in the samples; and (ii) select T. gondii oocysts. Toxoplasma gondii oocyst detection and an estimation of their numbers was performed by conventional PCR and real time qPCR, using specific primers for a 183-bp sequence of the T. gondii repetitive DNA region. All PCR-positive DNA samples were purified and sequenced. Restriction enzyme digestion with EcoRV endonuclease confirmed the presence of the T. gondii DNA fragment. In addition, the presence of the parasite was observed by fluorescent microscopy, taking advantage of the oocysts autofluorescence under UV light., Results: Forty percent of the analysed samples (95% CI: 25.5-56.5%) presented the expected PCR and digested DNA fragments. These fragments were confirmed by sequencing. Microscopic autofluorescence supported the presence of T. gondii-like oocysts. The estimated mean (± SE) oocyst concentration was 23.5 ± 12.1 oocysts/g, with a range of 0.6-179.9 oocysts/g., Conclusions: Our findings provide relevant evidence of contamination of fresh vegetables and berry fruits with T. gondii oocysts.

Bactrocera dorsalis (Hendel) and Ceratitis capitata (Wiedemann) are highly polyphagous fruit fly species and important pests of commercial fruit in regions of the world where they are present. In South Africa, B. dorsalis is now established in the north and northeastern parts of the country. B. dorsalis is currently absent in other parts of the country including the Western Cape Province which is an important area for the production of deciduous fruit. C. capitata is widespread in South Africa and is the dominant pest of deciduous fruit. The demographic parameters of B. dorsalis and C. capitata on four deciduous fruit types Prunus persica (L.) Batsch, Prunus domestica L., Malus domestica Borkh. and Pyrus communis L. were studied to aid in predicting the potential population establishment and growth of B. dorsalis in a deciduous fruit growing environment. All deciduous fruit types tested were suitable for population persistence of both B. dorsalis and C. capitata. Development was fastest and survival highest on nectarine for both species. B. dorsalis adults generally lived longer than those of C. capitata, irrespective of the fruit types that they developed from. B. dorsalis had a higher net reproductive rate (Ro) on all deciduous fruit tested compared to C. capitata. However, the intrinsic rate of population increase was estimated to be higher for C. capitata than for B. dorsalis on all fruit types tested primarily due to C. capitata's faster generation time. Provided abiotic conditions are optimal, B. dorsalis would be able to establish and grow in deciduous fruit growing areas.