Organic mulching modulated native populations of entomopathogenic nematode in vineyard soils differently depending on its potential to control outgrowth of their natural enemies.

The entomopathogenic nematodes (EPNs) are biological control agents that are widespread in crop soils. However, traditional agricultural management practices such as cultivation and agrochemical usage can alter the soil balance that enables their occurrence and activity. Alternative strategies like mulching are commonly employed to prevent weed growth, enhance below-ground biodiversity by improving soil, organic matter content, fertility, and moisture. We hypothesized that organic mulches would favor biotic conditions for nematofauna development in crop soil, including EPNs, compared to herbicide application or tillage. Traditional (insect baits) and molecular (qPCR analysis) tools were used in this study to assess the abundance and activity of native EPNs, and the abundance of potential natural enemies, such as free-living nematode (FLN) competitors, nematophagous fungi (NF), and ectoparasitic bacteria, in soils managed with different organic mulches or traditional practices. As a model agroecosystem, we selected the vineyard, one of the most intensively managed crop systems. We compared mulches of grape pruning debris (GPD-M), straw (Str-M), and spent mushroom compost (SMC-M) in two commercial vineyards, which employed either integrated or organic pest and disease management. Following a completely randomized design, we retrieved two composite samples per plot (n = 3 per treatment in each vineyard) in April, June, and October 2020. Numbers of EPNs and selected members of their soil food web were higher in the organic than the integrated managed vineyard. Supporting our hypothesis, organic mulching overall favored nematode occurrence in both vineyards. We found higher NF abundance for Str-M, and GPD-M in the organic vineyard, which plausibly explained the lower EPN activity and occurrence compared to SMC-M in both vineyards. We conclude that the organic mulches can provide appropriate conditions for increasing nematofauna numbers but, depending on the mulch type, may also adversely affect EPNs by increasing their natural enemies. Our findings highlight the need to explore alternative farming practices to unravel complex biotic interactions that affect beneficial soil organisms in agroecosystems.

The silk of gorse spider mite Tetranychus lintearius represents a novel natural source of nanoparticles and biomaterials.

Spider mites constitute an assemblage of well-known pests in agriculture, but are less known for their ability to spin silk of nanoscale diameters and high Young's moduli. Here, we characterize silk of the gorse spider mite Tetranychus lintearius, which produces copious amounts of silk with nano-dimensions. We determined biophysical characteristics of the silk fibres and manufactured nanoparticles and biofilm derived from native silk. We determined silk structure using attenuated total reflectance Fourier transform infrared spectroscopy, and characterized silk nanoparticles using field emission scanning electron microscopy. Comparative studies using T. lintearius and silkworm silk nanoparticles and biofilm demonstrated that spider mite silk supports mammalian cell growth in vitro and that fluorescently labelled nanoparticles can enter cell cytoplasm. The potential for cytocompatibility demonstrated by this study, together with the prospect of recombinant silk production, opens a new avenue for biomedical application of this little-known silk.

Effects of Ground Cover Management on Insect Predators and Pests in a Mediterranean Vineyard.

Conservative techniques, such as ground cover management, could help promote viticulture sustainability, which is a goal of conservation biological control, by providing shelter and food sources for predatory insects. A field experiment was conducted in a Mediterranean vineyard to evaluate ground cover management impacts on predatory insect and potential grapevine pest abundance and diversity, both on the ground and in the grapevine canopy. Three different ground cover management techniques (tillage, spontaneous cover and flower-driven cover) were tested for two years (2016 and 2017). Overall, the ground cover management significantly affected the abundance of important epigeal predators, of which carabids, forficulids and staphylinids were the most captured. The carabid abundances under both the cover crop treatments were found to be approximately three times higher compared with that under the tillage treatment. In contrast, the canopy insect abundance in the vineyard was similar among the treatments for both the predators and the potential grapevine pest species. These results indicate that cover crop vegetation can be used in vineyards to enhance predatory insect abundance and may improve agroecosystem resilience.

Scavenging behavior and interspecific competition decrease offspring fitness of the entomopathogenic nematode Steinernema feltiae.

Entomopathogenic nematodes (EPNs) are well-studied biocontrol agents of soil-dwelling arthropod pests. The insecticidal efficiency of EPNs is modulated by food web dynamics. EPNs can reproduce in freeze-killed insect larvae, even in competition with free-living bacterivorous nematodes (FLBNs) in the genus Oscheius. The objective of this study was to assess the efficiency of EPNs as scavengers when competing with free-living saprophagous nematodes and fungi, and to determine the possible impact on subsequent EPN offspring fitness. Live and freeze-killed larvae of Galleria mellonella were used to evaluate the reproduction rate and progeny fitness of two EPN species, Heterorhabditis bacteriophora and Steinernema feltiae, applied individually or combined with the FLBN species Oscheius onirici or Pristionchus maupasi, or Aspergillus flavus, an opportunistic saprophytic fungus. We hypothesized that (1) EPN scavenging behaviors previously observed (for H. megidis and S. kraussei) apply to other EPN species, (2) infective juveniles (IJs) emerging from freeze-killed larvae will display reduced pathogenicity and reproduction, and (3) fitness reduction will be amplified by exposure to other organisms competing for the resources. The reproduction rate of S. feltiae was lower in freeze-killed larvae than in larvae infected and killed by the nematode, whereas H. bacteriophora failed to reproduce as a scavenger. The S. feltiae F1 IJs that emerged from freeze-killed larvae exhibited lower pathogenicity rates than IJs resulting from entomopathogenic activity, and also lower reproductive rates if they experienced high FLBN competitive pressure during development. This study illustrates that scavenging is a suboptimal alternative pathway for EPNs, especially in the face of scavenger competition, even though it provides a means for some EPN species to complete their life-cycle.

Mathematical Models for Predicting Development of Orius majusculus (Heteroptera: Anthocoridae) and Its Applicability to Biological Control.

Complete development of Orius majusculus Reuter (Heteroptera: Anthocoridae) at nine constant temperatures, between 12 and 34°C, was evaluated under laboratory conditions. The maximum developmental period of 90.75 d occurred at 12°C, whereas the minimum of 11.34 d occurred at 30°C. From 30 to 34°C, the developmental period increased to 13.50 d. Between 21 and 33°C the survival rate was more than 80%. The optimal temperature when considering developmental rate and survival was between 24 and 30°C. At constant temperatures, four models were developed, one of which was linear and three nonlinear (Logan type III, Lactin, and Brière). All models were validated under field conditions and diel temperature variations. The values of the adjusted determination coefficients of the linear (>0.77) and nonlinear models (>0.93) were high. The thermal requirement for complete development, from egg to adult, was 284.5 degree-days (DD). In all nonlinear models, elevated levels of accuracy (≥90.31%) in field validation were also obtained, especially in the Brière model. With the results obtained herein, the optimization of O. majusculus mass rearing, its ideal use, and field management in biological control strategies can be improved.

Comparative genome-wide transcriptome analysis of Vitis vinifera responses to adapted and non-adapted strains of two-spotted spider mite, Tetranyhus urticae.

BACKGROUND: The two-spotted spider mite, Tetranychus urticae, is an extreme generalist plant pest. Even though mites can feed on many plant species, local mite populations form host races that do not perform equally well on all potential hosts. An acquisition of the ability to evade plant defenses is fundamental for mite's ability to use a particular plant as a host. Thus, understanding the interactions between the plant and mites with different host adaptation status allows the identification of functional plant defenses and ways mites can evolve to avoid them. RESULTS: The grapevine genome-wide transcriptional responses to spider mite strains that are non-adapted and adapted to grapevine as a host were examined. Comparative transcriptome analysis of grapevine responses to these mite strains identified the existence of weak responses induced by the feeding of the non-adapted strain. In contrast, strong but ineffective induced defenses were triggered upon feeding of the adapted strain. A comparative meta-analysis of Arabidopsis, tomato and grapevine responses to mite feeding identified a core of 36 highly conserved genes involved in the perception, regulation and metabolism that were commonly induced in all three species by mite herbivory. CONCLUSIONS: This study describes the genome-wide grapevine transcriptional responses to herbivory of mite strains that differ in their ability to use grapevine as a host. It raises hypotheses whose testing will lead to our understanding of grapevine defenses and mite adaptations to them.

Effects of Kaolin on Lobesia botrana (Lepidoptera: Tortricidae) and Its Compatibility With the Natural Enemy, Trichogramma cacoeciae (Hymenoptera: Trichogrammatidae).

Lobesia botrana (Denis and Schiffermüller) (Lepidoptera: Tortricidae) is an important grapevine pest in Europe recently encountered in America. Trichogramma cacoeciae Marchal (Hymenoptera: Trichogrammatidae) is amongst the most effective parasitoids for Lepidopteran species. Studies to evaluate the effect of kaolin, an inert, nontoxic mineral, on oviposition, egg hatch, and neonate mortality of these species were carried out. Efficacy on L. botrana neonate larvae, oviposition, and egg hatch was evaluated. Effects of kaolin on parasitism and emergence of T. cacoeciae from L. botrana and Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae) eggs were also evaluated. Lobesia botrana egg hatch and oviposition rates were reduced, and neonate larvae mortality was significantly greater in kaolin-treated arenas and when included in synthetic neonate larvae diet. Kaolin had no effect on T. cacoeciae parasitism in both hosts. There was only a slight but statistically insignificant effect on T. cacoeciae progeny emergence from L. botrana eggs and no effect from E. kuehniella. The results involving reductions in L. botrana oviposition and egg hatch and increase in larval mortality with kaolin suggest this compound may contribute to reduction in population densities and can be considered in rational integrated pest management strategies for L. botrana. Due to the laboratory results presented on parasitoid emergence, even though field bioassays would give a more exhaustive evaluation, it appears kaolin can be compatible with T. cacoeciae in L. botrana management.

Optimizing Nesidiocoris tenuis (Hemiptera: Miridae) as a biological control agent: mathematical models for predicting its development as a function of temperature.

For optimal application of Nesidiocoris tenuis as a biological control agent, adequate field management and programmed mass rearing are essential. Mathematical models are useful tools for predicting the temperature-dependent developmental rate of the predator. In this study, the linear model and nonlinear models Logan type III, Lactin and Brière were estimated at constant temperatures and validated at alternating temperatures and under field conditions. N. tenuis achieved complete development from egg to adult at constant temperatures between 15 and 35°C with high survivorship (>80%) in the range 18-32°C. The total developmental time decreased from a maximum at 15°C (76.74 d) to a minimum at 33°C (12.67 d) and after that, increased to 35°C (13.98 d). Linear and nonlinear developmental models all had high accuracy (R a 2 >0.86). The maximum developmental rate was obtained between 31.9°C (Logan type III and Brière model for N1) and 35.6°C (for the egg stage in the Brière model). Optimal survival and the highest developmental rate fell within the range 27-30°C. The field validation revealed that the Logan type III and Lactin models offered the best predictions (95.0 and 94.5%, respectively). The data obtained on developmental time and mortality at different temperatures are useful for mass rearing this predator, and the developmental models are valuable for using N. tenuis as a biological control agent.

Acute and reproductive effects of Align, an insecticide containing azadirachtin, on the grape berry moth, Lobesia botrana.

Azadirachtin, derived from the neem tree, Azadirachta indica A. Juss (Sapindales: Meliaceae), seems promising for use in integrated pest management programs to control a variety of pest species. A commercial formulation of azadirachtin, Align, has been evaluated against different developmental stages of the European grape berry moth, Lobesia botrana Denis and Schiffermüller (Lepidoptera: Tortricidae). When administered orally, Align reduced the fecundity and fertility of adults treated with 1, 5, and 10 mg litre(-1). At the highest doses, fecundity and fertility were zero, but longevity was not affected. An LC(50) of 231.5 mg litre(-1) was obtained when Align was sprayed on eggs less than 1 day old. Hatching of all egg classes was significantly reduced, and this reduction was more pronounced for eggs less than 24 h old. LC(50) values of 2.1 mg litre(-1) for first instars and 18.7 mg litre(-1) for third instars were obtained when Align was present in the diet. Larvae reared on a diet containing different concentrations of Align did not molt into adults at the highest concentrations (0.3, 0.6, 1.2), and 50% molted at the lowest concentration (0.15). Phenotypic effects included inability to molt properly and deformities. The combination of acute toxicity and low, effective concentrations of Align observed in this study could lead to the inclusion of insecticides containing azadirachtin in integrated management programs against this pest.

Effects of Lobesia botrana (Lepidoptera: Tortricidae) egg age, density, and UV treatment on parasitism and development of Trichogramma cacoeciae (Hymenoptera: Trichogrammatidae).

Integrated pest management (IPM) supports the integration of all rational control methods available. Biological control is one of those control methods. Thus, the influence of Lobesia botrana Den. and Schiff. egg age, density, and sterilization by UV light on parasitism and progeny development of a Trichogramma cacoeciae Marchal strain from La Rioja, Spain, was studied. Understanding this influence is important for the development of biological control programs. UV-killed eggs are widely used for mass rearing Trichogramma. Trichogramma cacoeciae preferred young eggs (0-72 h old), and the largest number of progeny were produced from these eggs, especially from 48- to 72-h-old eggs. The relationship between the number of eggs parasitized by T. cacoeciae and its density (within the range studied) was linear (R (2) adjusted = 0.93), with constant values (72.2-93.7%) for discovery rate (an estimator of searching capacity). UV treatment did not affect the number of progeny produced or parasitism tendency and was thus a helpful tool for the mass-rearing process. Based on its response to egg age and density, our T. cacoeciae strain showed potential for control of L. botrana.

Effects of methoxyfenozide on Lobesia botrana Den & Schiff (Lepidoptera: Tortricidae) egg, larval and adult stages.

The effect of the non-steroidal ecdysone agonist methoxyfenozide was evaluated against different developmental stages of the grape berry moth, Lobesia botrana Dennis & Schiffermuller (Lep, Tortricidae). Methoxyfenozide administered orally reduced the fecundity and fertility of adults treated with 1, 5 and 10 mg litre(-1); longevity was not affected. An LC(50) value of 4.5 mg litre(-1) was obtained when applied to eggs of less than 1 day old. Surface treatment was more effective than when applied by spraying. Administered into the diet, methoxyfenozide had a larvicidal effect; older larvae were more susceptible than younger larvae, with LC(50) values of 0.1 mg litre(-1) for L(1), 0.04 for L(3) and 0.02 for L(5). Larvae treated with sub-lethal doses throughout their lives did not emerge as adults at the highest doses (0.08, 0.04, 0.02 and 0.01 mg litre(-1)), with 65% and 40% emergence occurring for the lowest (0.005 and 0.0025 mg litre(-1)). Mortality occurred only in the larval stage.

Effects of azadirachtin on the two-spotted spider mite, Tetranychus urticae (Acari: Tetranychidae).

Effects of sublethal azadirachtin exposure to the biological performance of Tetranychius urticae Koch was studied under laboratory conditions. Bioassay was used to asses the effect of different concentrations of azadirachtin on longevity, fecundity, fertility, and offspring development. Azadirachtin (64 and 128 ppm) affected fecundity and mortality but had no effect on fertility and offspring development. A subsequent life-table study with 80 ppm of azadirachtin found that the compound caused a reduction of 50% in survival to adult stage. The peak of reproduction was reached at 5 days causing a decrease in the mean fecundity to almost eight times than of untreated females. The net reproductive rate (R0), the intrinsic rate of increase (r(m)), and the finite rate of increase (lambda) of treated females were lower. Treatment showed a negative value of r(m), resulting in a declining population. These results suggest that azadirachtin could be incorporated in Integrated Pest Management (IPM) programmes of T. urticae.