Large-scale insect rearing can be subjected to microbial infections, leading to serious economic losses. When possible, the use of antibiotics is to be avoided for insects intended as feed or food ...and new strategies to preserve the health of the farmed insects are required. The effectiveness of insect immune system depends on several factors, including the nutritional composition of the diet. The possibility to modulate immune responses through the diet is currently a topic of great interest from an application point of view. We evaluated the effect of two different diets on the survival rate and gene expression of the antimicrobial peptide Tenecin 3 in uninfected and
-infected
L. larvae. A wheat bran diet, mixed 50% with brewers' spent grains, could positively influence the expression of Tenecin 3 gene when uninfected
is allowed to develop on such a substrate from early larval stages. Even if, in our trial, the diet with added brewers' spent grains could not reduce the mortality of the larvae infected with
, higher transcriptional levels of the antifungal peptide were observed in insects fed this diet, depending on the timing of diet administration.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
Some pathogenic phloem‐limited bacteria are a major threat for worldwide agriculture due to the heavy economic losses caused to many high‐value crops. These disease agents – phytoplasmas, ...spiroplasmas, liberibacters, and Arsenophonus‐like bacteria – are transmitted from plant to plant by phloem‐feeding Hemiptera vectors. The associations established among pathogens and vectors result in a complex network of interactions involving also the whole microbial community harboured by the insect host. Interactions among bacteria may be beneficial, competitive, or detrimental for the involved microorganisms, and can dramatically affect the insect vector competence and consequently the spread of diseases. Interference is observed among pathogen strains competing to invade the same vector specimen, causing selective acquisition or transmission. Insect bacterial endosymbionts are another pivotal element of interactions between vectors and phytopathogens, because of their central role in insect life cycles. Some symbionts, either obligate or facultative, were shown to have antagonistic effects on the colonization by plant pathogens, by producing antimicrobial substances, by stimulating the production of antimicrobial substances by insects, or by competing for host infection. In other cases, the mutual exclusion between symbiont and pathogen suggests a possible detrimental influence on phytopathogens displayed by symbiotic bacteria; conversely, examples of microbes enhancing pathogen load are available as well. Whether and how bacterial exchanges occurring in vectors affect the relationship between insects, plants, and phytopathogens is still unresolved, leaving room for many open questions concerning the significance of particular traits of these multitrophic interactions. Such complex interplays may have a serious impact on pathogen spread and control, potentially driving new strategies for the containment of important diseases.
Phloem‐limited bacterial phytopathogens, transmitted among plants by phloem‐feeding Hemiptera vectors, cause heavy losses in agriculture. Associations of pathogens and vectors form complex networks of interactions involving also the microbial community harboured by the insect host; bacterial interactions may have beneficial or detrimental influences on a vector's efficacy. How such complex interplays affect insect‐plant‐bacteria interactions is still unresolved; however, their impact on pathogen spread and control could drive new strategies for the containment of important diseases.
The black soldier fly (BSF),
L. (Diptera: Stratiomyidae), has a great bioconversion potential and ability to develop on diverse substrates. Although the use of catering waste and food by-products ...containing meat and fish would reduce the footprint of the insect sector, to date, in Europe, their use is still facing legal obstacles for insects as food and feed. Since a major request from the EU insect sector is to diversify the spectrum of allowed substrates, and considering that variations in diet composition could influence insect immune responses, we evaluated the impact of different catering wastes on BSF prepupae immunity. Insects were reared on five diets: one based on feed for laying hens and four based on catering waste containing (i) fruits and vegetables; (ii) fruits, vegetables and bread; (iii) fruit, vegetables, bread and dairy products; (iv) fruits, vegetables, bread, meat and fish. The gene expression of two antimicrobial peptides (AMPs), one defensin and one cecropin, was assessed. Moreover, the hemolymph inhibitory activity against
DH5α and
HI55 was evaluated using diffusion assays in solid media. The up-regulation of both AMPs' encoding genes was observed in insects fed a bread-added and dairy product-added diet. All hemolymph samples showed inhibitory activity against both bacteria, affecting the colony size and number. The obtained results show how catering waste positively influences the
immune system. The possibility of modulating AMP expression levels through the diet opens up new perspectives in the management of insect health in mass rearings.
Several insects are known as vectors of a wide range of animal and human pathogens causing various diseases. However, they are also a source of different substances, such as the Antimicrobial ...Peptides (AMPs), which can be employed in the development of natural bioactive compounds for medical, veterinary and agricultural applications. It is well known that AMP activity, in contrast to most classical antibiotics, does not lead to the development of natural bacterial resistance, or at least the frequency of resistance is considered to be low. Therefore, there is a strong interest in assessing the efficacy of the various peptides known to date, identifying new compounds and evaluating possible solutions in order to increase their production. Moreover, implementing AMP modulation in insect rearing could preserve insect health in large-scale production. This review describes the current knowledge on insect AMPs, presenting the validated ones for the different insect orders. A brief description of their mechanism of action is reported with focus on proposed applications. The possible effects of insect diet on AMP translation and synthesis have been discussed.
The transmission of phytoplasmas is the result of an intricate interplay involving pathogens, insect vectors and host plants. Knowledge of the vector's competence during its lifespan allows us to ...define more sustainable well-timed control strategies targeted towards the most worrisome life stages. We investigated the temporal dynamics of '
Phytoplasma mali' load in
in the different developmental stages in Northwest Italy. The phytoplasma load in the vector was evaluated in overwintering adults, nymphs and newly emerged adults after different acquisition access periods. Moreover, we followed the multiplication of the phytoplasma during the aestivation and the overwintering period on conifers. Our results confirmed the ability of remigrants to retain the phytoplasma until the end of winter. We also highlighted the high acquisition efficiency and vector competence, based on phytoplasma load, of nymphs and newly emerged adults. Therefore, particular attention should be paid to the management of overwintered
as soon as they return to the orchards, but also to newly emerged adults, particularly in orchards with a high infection rate and when the migration to conifers is delayed.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
Insect immunity is a crucial process in interactions between host and microorganisms and the presence of pathogenic, commensal, or beneficial bacteria may result in different immune responses. In ...Hemiptera vectors of phytoplasmas, infected insects are amenable to carrying high loads of phytopathogens, besides hosting other bacterial affiliates, which have evolved different strategies to be retained; adaptation to host response and immunomodulation are key aspects of insect-symbiont interactions. Most of the analyses published to date has investigated insect immune response to pathogens, whereas few studies have focused on the role of host immunity in microbiota homeostasis and vectorial capacity. Here the expression of immune genes in the leafhopper vector of phytoplasmas
was investigated following exposure to
symbiotic bacteria, previously demonstrated to affect phytoplasma acquisition by leafhoppers. The expression of four genes related to major components of immunity was measured, i.e., defensin, phenoloxidase, kazal type 1 serine protease inhibitor and Raf, a component of the Ras/Raf pathway. The response was separately tested in whole insects, midguts and cultured hemocytes. Healthy individuals were assessed along with specimens undergoing early- and late-stage phytoplasma infection. In addition, the adhesion grade of
strains was examined to assess whether symbionts could establish a physical barrier against phytoplasma colonization. Our results revealed a specific activation of Raf in midguts after double infection by
and flavescence dorée phytoplasma. Increased expression was observed already in early stages of phytoplasma colonization. Gut-specific localization and timing of Raf activation are consistent with the role played by
in limiting phytoplasma acquisition by
, supporting the involvement of this gene in the anti-pathogen activity. However, limited attachment capability was found for
under
experimental conditions, suggesting a minor contribution of physical phytoplasma exclusion from the vector gut wall. By providing evidence of immune modulation played by
, these results contribute to elucidating the molecular mechanisms regulating interference with phytoplasma infection in
. The involvement of Raf suggests that in the presence of reduced immunity (reported in Hemipterans), immune genes can be differently regulated and recruited to play additional functions, generally played by genes lost by hemipterans.
Over the past few years, there has been an increasing interest in the development of alternative pest control strategies to reduce environmental impact. In this contest, exclusion nets have been ...evaluated as a sustainable alternative to pesticides. In this study, the use of a photoselective exclusion net was investigated in semi-field conditions as a potential strategy to protect nectarine orchards from different pests (i.e., fruit moths,
and
) in NW Italy. The presence and abundance of pest populations inside and outside the net, as well as the damage they caused on fruits, were evaluated. Moreover, any possible effects of the net on beneficial arthropods, postharvest rots and fruit quality and nutraceutical parameters were considered. The exclusion net significantly reduced pest populations. At harvest, fruit damage caused by
and
in netted plots was reduced up to 90% and to 78%, respectively, compared with insecticide-treated plots. The exclusion net allowed the production of healthier fruits with a strong reduction of insecticide treatments (up to seven less) and of their related costs without any negative impact on postharvest rots, neither fruit quality nor nutraceutical properties.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
One of the most destructive diseases affecting grapevine in Europe is caused by Flavescence Dorée phytoplasma (FDp), which belongs to the 16Sr-V group and is a European Union quarantine pathogen. ...Although many molecular techniques such as loop-mediated isothermal amplification (LAMP) are widely used for the rapid detection of FDp in infected grapevine plants, there is no developed isothermal amplification assay for FDp detection in the insect vectors that are fundamental for the spread of the disease. For this reason, a simple in-field real-time LAMP protocol was optimized and developed for the specific detection of FDp in the insect vector Scaphoideus titanus. The LAMP assay was optimized to work with crude insect extracts obtained by manually shaking a single insect in a buffer for 5 min. Such a simple, sensitive, specific, economic, and user-friendly LAMP assay allowed the detection of FDp in S. titanus in less than half an hour, directly in the field. The developed insect tissue preparation procedure, combined with the LAMP protocol, promptly revealed the presence of FDp in infected S. titanus directly in the vineyards, allowing for monitoring of the spread of the pathogen in the field and to apply timely strategies required for the mandatory control of this pathogen.
One emerging disease of grapevine in Europe is Bois noir (BN), a phytoplasmosis caused by "Candidatus Phytoplasma solani" and spread in vineyards by the planthopper Hyalesthes obsoletus (Hemiptera: ...Cixiidae). Here we present the first full characterization of the bacterial community of this important disease vector collected from BN-contaminated areas in Piedmont, Italy. Length heterogeneity PCR and denaturing gradient gel electrophoresis analysis targeting the 16S rRNA gene revealed the presence of a number of bacteria stably associated with the insect vector. In particular, symbiotic bacteria detected by PCR with high infection rates in adult individuals fell within the "Candidatus Sulcia muelleri" cluster in the Bacteroidetes and in the "Candidatus Purcelliella pentastirinorum" group in the Gammaproteobacteria, both previously identified in different leafhoppers and planthoppers. A high infection rate (81%) was also shown for another symbiont belonging to the Betaproteobacteria, designated the HO1-V symbiont. Because of the low level of 16S rRNA gene identity (80%) with the closest relative, an uncharacterized symbiont of the tick Haemaphysalis longicornis, we propose the new name "Candidatus Vidania fulgoroideae." Other bacterial endosymbionts identified in H. obsoletus were related to the intracellular bacteria Wolbachia pipientis, Rickettsia sp., and "Candidatus Cardinium hertigii." Fluorescent in situ hybridization coupled with confocal laser scanning microscopy and transmission electron microscopy showed that these bacteria are localized in the gut, testicles, and oocytes. As "Ca. Sulcia" is usually reported in association with other symbiotic bacteria, we propose that in H. obsoletus, it may occur in a bipartite or even tripartite relationship between "Ca. Sulcia" and "Ca. Purcelliella," "Ca. Vidania," or both.
With the growing interest in insect rearing for feed and food use, there has been an increased focus on the health of the insects for both farm business aspects and the safety of end consumers. ...Recent research has highlighted how the insect immune system can be modulated by the diet. We studied the survival of yellow mealworm beetle larvae infected with an entomopathogenic fungus and the gene expression of a specific antimicrobial peptide with antifungal action in insects fed two different diets. In our trials, the partial substitution of a wheat bran-based diet by brewers’ spent grain, characterized by a high protein value, did not increase the survival rate of infected larvae but increased the expression of the antifungal peptide, depending on the timing of administration. The possibility of modulating farmed insect health through the diet, using agri-food byproducts, opens interesting new perspectives within a circular economy scenario. Large-scale insect rearing can be subjected to microbial infections, leading to serious economic losses. When possible, the use of antibiotics is to be avoided for insects intended as feed or food and new strategies to preserve the health of the farmed insects are required. The effectiveness of insect immune system depends on several factors, including the nutritional composition of the diet. The possibility to modulate immune responses through the diet is currently a topic of great interest from an application point of view. We evaluated the effect of two different diets on the survival rate and gene expression of the antimicrobial peptide Tenecin 3 in uninfected and Beauveria bassiana-infected Tenebrio molitor L. larvae. A wheat bran diet, mixed 50% with brewers’ spent grains, could positively influence the expression of Tenecin 3 gene when uninfected T. molitor is allowed to develop on such a substrate from early larval stages. Even if, in our trial, the diet with added brewers’ spent grains could not reduce the mortality of the larvae infected with B. bassiana, higher transcriptional levels of the antifungal peptide were observed in insects fed this diet, depending on the timing of diet administration.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK