Kill rate as a tool in efficiency evaluation of Neoseiulus californicus (Acari: Phytoseiidae) mass reared on factitious food.

The predatory mites of the Phytoseiidae family are crucial biological control agents widely utilized in biological pest management targeting phytophagous mites and insects. Key factors in these control strategies are that phytoseiids must be able to find their main target prey and to maintain high populations and efficacy. To reduce expenses and time-consuming production methods of mass rearing of phytoseiids, pollen and other factitious (i.e., non-natural/nontarget) hosts need to be present as an alternative food for predatory mite populations. The mass-rearing possibilities of these predators on alternative food sources, such as astigmatid mites (i.e., house and stored mites) and pollen, must be evaluated not only by the cost of rearing settings but on the basis of their efficiency maintenance in killing prey. The pest kill rate (km) is the average daily lifetime killing of the pest by the natural enemy under consideration and can represent a useful indicator for phytoseiids efficacy to rank them as BCAs on the basis of their effective killing/predation on target prey. In this study, we evidenced that 2 astigmatid mites, Glycyphagus domesticus (De Geer) and Lepidoglyphus destructor (Schrank), and Quercus ilex L. pollen can be successfully adopted as substitute food sources for mass rearing of the phytoseiid Neoseiulus californicus (MgGregor); then, we determined that these populations of BCAs maintained a consistent km at new/first impact on the main target pest, Tetranychus urticae Koch.

Tenebrio molitor as a Simple and Cheap Preclinical Pharmacokinetic and Toxicity Model.

The progression of drugs into clinical phases requires proper toxicity assessment in animals and the correct identification of possible metabolites. Accordingly, different animal models are used to preliminarily evaluate toxicity and biotransformations. Rodents are the most common models used to preliminarily evaluate the safety of drugs; however, their use is subject to ethical consideration and elevated costs, and strictly regulated by national legislations. Herein, we developed a novel, cheap and convenient toxicity model using Tenebrio molitor coleoptera (TMC). A panel of 15 drugs-including antivirals and antibacterials-with different therapeutic applications was administered to TMC and the LD50 was determined. The values are comparable with those already determined in mice and rats. In addition, a TMC model was used to determine the presence of the main metabolites and in vivo pharmacokinetics (PK), and results were compared with those available from in vitro assays and the literature. Taken together, our results demonstrate that TMC can be used as a novel and convenient preliminary toxicity model to preliminarily evaluate the safety of experimental compounds and the formation of main metabolites, and to reduce the costs and number of rodents, according to 3R principles.

Genetic analysis of the tomato russet mite provides evidence of oligophagy and a widespread pestiferous haplotype.

Worldwide, the tomato russet mite (TRM), Aculops lycopersici (Eriophyidae), is a key pest on cultivated tomato in addition to infesting other cultivated and wild Solanaceae; however, basic information on TRM supporting effective control strategies is still lacking, mainly regarding its taxonomic status and genetic diversity and structure. As A. lycopersici is reported on different species and genera of host plants, populations associated with different host plants may constitute specialized cryptic species, as shown for other eriophyids previously considered generalists. The main aims of this study were to (i) confirm the TRM taxonomic unity of populations from different host plants and localities as well as the species' oligophagy, and (ii) to advance the understanding of TRM host relationship and invasion history. For this purpose, we evaluated the genetic variability and structure of populations from different host plants along crucial areas of occurrence, including the area of potential origin, based on DNA sequences of mitochondrial (cytochrome c oxidase subunit I) and nuclear (internal transcribed spacer, D2 28S) genomic regions. Specimens from South America (Brazil) and Europe (France, Italy, Poland, The Netherlands) were collected from tomato and other solanaceous species from the genera Solanum and Physalis. Final TRM datasets were composed of 101, 82 and 50 sequences from the COI (672 bp), ITS (553 bp) and D2 (605 bp) regions, respectively. Distributions and frequencies of haplotypes (COI) and genotypes (D2 and ITS1) were inferred; pairwise genetic distance comparisons, and phylogenetic analysis were performed, including Bayesian Inference (BI) combined analysis. Our results showed that genetic divergences for mitochondrial and nuclear genomic regions from TRM associated with different host plants were lower than those observed in other eriophyid taxa, confirming conspecificity of TRM populations and oligophagy of this eriophyid mite. Four haplotypes (cH) were identified from the COI sequences with cH1 being the most frequent, representing 90% of all sequences occurring in all host plants studied (Brazil, France, The Netherlands); the other haplotypes were present exclusively in Brazilian populations. Six variants (I) were identified from the ITS sequences: I-1 was the most frequent (76.5% of all sequences), spread in all countries and associated with all host plants, except S. nigrum. Just one D2 sequence variant was found in all studied countries. The genetic homogeneity among populations highlights the occurrence of a highly invasive and oligophagous haplotype. These results failed to corroborate the hypothesis that differential symptomatology or damage intensity among tomato varieties and solanaceous host plants could be due to the genetic diversity of the associated mite populations. The genetic evidence, along with the history of spread of cultivated tomato, corroborates the hypothesis of a South American origin of TRM.

Effects of soil erosion on agro-ecosystem services and soil functions: A multidisciplinary study in nineteen organically farmed European and Turkish vineyards.

This multidisciplinary research work evaluated the effects of soil erosion on grape yield and quality and on different soil functions, namely water and nutrient supply, carbon sequestration, organic matter recycling, and soil biodiversity, with the aim to understand the causes of soil malfunctioning and work out a proper strategy of soil remediation. Degraded areas in nineteen organically farmed European and Turkish vineyards resulted in producing significantly lower amounts of grapes and excessive concentrations of sugar. Plants suffered from decreased water nutrition, due to shallower rooting depth, compaction, and reduced available water capacity, lower chemical fertility, as total nitrogen and cation exchange capacity, and higher concentration of carbonates. Carbon storage and organic matter recycling were also depressed. The general trend of soil enzyme activity mainly followed organic matter stock. Specific enzymatic activities suggested that in degraded soils, alongside a general slowdown in organic matter cycling, there was a greater reduction in decomposition capacity of the most recalcitrant forms. The abundance of Acari Oribatida and Collembola resulted the most sensitive indicator of soil degradation among the considered microarthropods. No clear difference in overall microbial richness and evenness were observed. All indices were relatively high and indicative of rich occurrence of many and rare microbial species. Dice cluster analyses indicated slight qualitative differences in Eubacterial and fungal community compositions in rhizosphere soil and roots in degraded soils. This multidisciplinary study indicates that the loss of soil fertility caused by excessive earth movement before planting, or accelerated erosion, mainly affects water nutrition and chemical fertility. Biological soil fertility is also reduced, in particular the ability of biota to decompose organic matter, while biodiversity is less affected, probably because of the organic management. Therefore, the restoration of the eroded soils requires site-specific and intensive treatments, including accurately chosen organic matrices for fertilization, privileging the most easily decomposable. Restoring soil fertility in depth, however, remain an open question, which needs further investigation.

Defense suppression benefits herbivores that have a monopoly on their feeding site but can backfire within natural communities.

BACKGROUND: Plants have inducible defenses to combat attacking organisms. Hence, some herbivores have adapted to suppress these defenses. Suppression of plant defenses has been shown to benefit herbivores by boosting their growth and reproductive performance. RESULTS: We observed in field-grown tomatoes that spider mites (Tetranychus urticae) establish larger colonies on plants already infested with the tomato russet mite (Aculops lycopersici). Using laboratory assays, we observed that spider mites have a much higher reproductive performance on russet mite-infested plants, similar to their performance on the jasmonic acid (JA)-biosynthesis mutant def-1. Hence, we tested if russet mites suppress JA-responses thereby facilitating spider mites. We found that russet mites manipulate defenses: they induce those mediated by salicylic acid (SA) but suppress those mediated by JA which would otherwise hinder growth. This suppression of JA-defenses occurs downstream of JA-accumulation and is independent from its natural antagonist SA. In contrast, spider mites induced both JA- and SA-responses while plants infested with the two mite species together display strongly reduced JA-responses, yet a doubled SA-response. The spider mite-induced JA-response in the presence of russet mites was restored on transgenic tomatoes unable to accumulate SA (nahG), but russet mites alone still did not induce JA-responses on nahG plants. Thus, indirect facilitation of spider mites by russet mites depends on the antagonistic action of SA on JA while suppression of JA-defenses by russet mites does not. Furthermore, russet mite-induced SA-responses inhibited secondary infection by Pseudomonas syringae (Pst) while not affecting the mite itself. Finally, while facilitating spider mites, russet mites experience reduced population growth. CONCLUSIONS: Our results show that the benefits of suppressing plant defenses may diminish within communities with natural competitors. We show that suppression of defenses via the JA-SA antagonism can be a consequence, rather than the cause, of a primary suppression event and that its overall effect is determined by the presence of competing herbivores and the distinct palette of defenses these induce. Thus, whether or not host-defense manipulation improves an herbivore's fitness depends on interactions with other herbivores via induced-host defenses, implicating bidirectional causation of community structure of herbivores sharing a plant.

Morphological Markers for Cryopreservation in the Embryonic Development of Drosophila suzukii (Diptera: Drosophilidae).

Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) is an invasive pest recently reported in Europe whose spread into new areas has caused severe economic damage to many agricultural crops. There are serious concerns about the currently available chemical insecticides because of their low efficacy in controlling the species and their environmental impact; so, several studies have focused on environmentally safe strategies. The sterile insect technique (SIT), which requires colony maintenance in laboratory and production of large numbers of live animals, can be utilized in pest management programs and could be integrated with other control strategies if the potential risks associated with the rearing and maintenance of the insect line under laboratory conditions are given sufficient attention. In this regard, the ability to cryobiologically preserve such stocks would be of substantial value. Important prerequisites for long-term cryopreservation are determination of the embryonic stages, identification of specific embryonic stages, and knowledge of development time. This paper describes the main visible markers for the different stages of embryonic development and determines the timing of development at 25°C. D. suzukii embryogenesis lasts 23-25 h at 25°C and can be divided into 17 stages defined by specific morphological markers. The point at which 50% of embryos are at Stage 14 and 50% are at Stage 15, the most tolerant stages for cryopreservation treatment, as ascertained for Drosophila melanogaster Meigen in prior studies, is reached in 14-15 h. The efficiency of this procedure might be impaired by the retention of eggs in the oviducts, making it impossible to determine the stage of embryonic development for ∼25% of laid eggs.

Thyreophagus corticalis as a vector of hypovirulence in Cryphonectria parasitica in chestnut stands.

The natural spread of hypovirulence in Cryphonectria parasitica (Murr.) Barr. occurs in chestnut (Castanea sativa Mill) stands and orchards in Italy and other European countries, leading to spontaneous recovery of the diseased trees. Little is known about how hypovirulence spreads in chestnut stands but various corticolous mite species frequently detected on chestnut cankers could be one of the many factors playing a role in the spread. Artificial virulent cankers created in inoculation field tests and treated with Thyreophagus corticalis (Acari, Sarcoptiformes, Acaridae) raised on hypovirulent cultures showed similar growth to those treated with mycelia of the hypovirulent strain over 18 months of inoculation. Cultures re-isolated from virulent cankers treated with mites were found to contain hypovirus like those derived from pairings of virulent and hypovirulent strains. Viral dsRNA could be carried externally and/or ingested by mites from the hypovirulent mycelia and then transmitted to the mycelia of virulent strains, causing their conversion. In a laboratory study, all fecal pellets collected from mites reared on hypovirulent and virulent strains grown on semi-selective media gave rise to colonies of C. parasitica with similar morphological characters and virulence to the original cultures. Field inoculation of stump sprouts with the resulting colonies revealed that mite digestive tract passage did not alter the virulence of the studied strains. These results are of interest for the biological control of chestnut blight.

Molecular detection assay of the bud mite Trisetacus juniperinus on Cupressus sempervirens in nurseries of central Italy.

Trisetacus juniperinus (Nalepa) sensu Keifer (Acari: Eriophyoidea: Phytoptidae) causes irregular development of buds, shoot deformations and stunted growth of trees, resulting in a serious threat to nurseries and young stands of Cupressus sempervirens L. (Mediterranean cypress). Recently, some cypress clones selected for their resistance to the fungal canker agent Seiridium cardinale (Wag.) have shown high susceptibility to the mite. Considering its tiny body, its hidden lifestyle inside the buds and the probable occurrence of other species (the vagrant Epitrimerus cupressi (Keifer) is common on the Mediterranean cypress in Italy), detection and monitoring of T. juniperinus require taxonomic expertise and are often time-consuming and challenging before serious damage is discernible. In the present study, a rapid, cost-effective PCR-based method was developed and validated to detect T. juniperinus on cypresses. The cytochrome c oxidase subunit I gene was amplified with degenerate and specific primers, but the latter were the only ones able to discriminate between T. juniperinus and E. cupressi. PCR products distinguished the two species both in a pool of individuals in a mixed population of both species and in single individuals, indicating the sensitivity of the detection method. PCR-RFLP (restriction fragment length polymorphism) by means of XmnI and XbaI endonucleases separated the two species. Furthermore, a washing-sieving protocol was used to make mite collection from the tree sample faster and simpler; this procedure did not interfere with the molecular detection of the species. The possibility of the routine use of this assay to monitor quarantine eriophyoids infesting plant material is discussed.

Molecular Methods for the Simultaneous Detection of Tomato Fruit Blotch Virus and Identification of Tomato Russet Mite, a New Potential Virus-Vector System Threatening Solanaceous Crops Worldwide.

Tomato fruit blotch virus (ToFBV) (Blunervirus solani, family Kitaviridae) was firstly identified in Italy in 2018 in tomato plants that showed the uneven, blotchy ripening and dimpling of fruits. Subsequent High-Throughput Sequencing (HTS) analysis allowed ToFBV to be identified in samples collected in Australia, Brazil, and several European countries, and its presence in tomato crops was dated back to 2012. In 2023, the virus was found to be associated with two outbreaks in Italy and Belgium, and it was included in the EPPO Alert list as a potential new threat for tomato fruit production. Many epidemiologic features of ToFBV need to be still clarified, including transmission. Aculops lycopersici Massee (Acariformes: Eriophyoidea), the tomato russet mite (TRM), is a likely candidate vector, since high population densities were found in most of the ToFBV-infected tomato cultivations worldwide. Real-time RT-PCR tests for ToFBV detection and TRM identification were developed, also as a duplex assay. The optimized tests were then transferred to an RT-ddPCR assay and validated according to the EPPO Standard PM 7/98 (5). Such sensitive, reliable, and validated tests provide an important diagnostic tool in view of the probable threat posed by this virus-vector system to solanaceous crops worldwide and can contribute to epidemiological studies by simplifying the efficiency of research. To our knowledge, these are the first molecular methods developed for the simultaneous detection and identification of ToFBV and TRM.

Eriophyid Mites in Classical Biological Control of Weeds: Progress and Challenges.

A classical biological control agent is an exotic host-specific natural enemy, which is intentionally introduced to obtain long-term control of an alien invasive species. Among the arthropods considered for this role, eriophyid mites are likely to possess the main attributes required: host specificity, efficacy, and long-lasting effects. However, so far, only a few species have been approved for release. Due to their microscopic size and the general lack of knowledge regarding their biology and behavior, working with eriophyids is particularly challenging. Furthermore, mites disperse in wind, and little is known about biotic and abiotic constraints to their population growth. All these aspects pose challenges that, if not properly dealt with, can make it particularly difficult to evaluate eriophyids as prospective biological control agents and jeopardize the general success of control programs. We identified some of the critical aspects of working with eriophyids in classical biological control of weeds and focused on how they have been or may be addressed. In particular, we analyzed the importance of accurate mite identification, the difficulties faced in the evaluation of their host specificity, risk assessment of nontarget species, their impact on the weed, and the final steps of mite release and post-release monitoring.