Production of the yolk protein precursor vitellogenin is mediated by target of rapamycin (TOR) in the soft tick Ornithodoros moubata (Acari: Argasidae).

Initiation of vitellogenesis by blood feeding is essential for egg maturation in ticks. Nutrients derived from the blood meal are utilized by female ticks to synthesize the yolk protein precursor vitellogenin (Vg). Engorged Ornithodoros moubata ticks can synthesize Vg whether mated or virgin, thus O. moubata is an excellent model for studying the relative roles of blood feeding and mating in tick vitellogenesis. Injection of rapamycin into engorged O. moubata resulted in a reduction of ovarian growth and yolk accumulation in the oocytes of mated females. OmVg expression in the midgut and fat body and protein concentrations in the hemolymph significantly decreased in mated ticks after injection with rapamycin, indicating that inhibition of the nutrient-sensing target of rapamycin (TOR) pathway disrupts egg maturation at the levels of Vg expression and synthesis. These results suggest that the TOR-signaling pathway induces vitellogenesis in response to nutritional stimulation after a blood meal in O. moubata and is functionally independent of the mating-induced pathway.

Semisynthesis of an evasin from tick saliva reveals a critical role of tyrosine sulfation for chemokine binding and inhibition.

Blood-feeding arthropods produce antiinflammatory salivary proteins called evasins that function through inhibition of chemokine-receptor signaling in the host. Herein, we show that the evasin ACA-01 from the Amblyomma cajennense tick can be posttranslationally sulfated at two tyrosine residues, albeit as a mixture of sulfated variants. Homogenously sulfated variants of the proteins were efficiently assembled via a semisynthetic native chemical ligation strategy. Sulfation significantly improved the binding affinity of ACA-01 for a range of proinflammatory chemokines and enhanced the ability of ACA-01 to inhibit chemokine signaling through cognate receptors. Comparisons of evasin sequences and structural data suggest that tyrosine sulfation serves as a receptor mimetic strategy for recognizing and suppressing the proinflammatory activity of a wide variety of mammalian chemokines. As such, the incorporation of this posttranslational modification (PTM) or mimics thereof into evasins may provide a strategy to optimize tick salivary proteins for antiinflammatory applications.

Antiproliferative and antiangiogenic effect of Amblyomma sculptum (Acari: Ixodidae) crude saliva in endothelial cells in vitro.

Neovascularization, a process that includes vasculogenesis and angiogenesis, may be a physiological or pathologic event, but in any cases the phenomenon is related to the formation of vascular net and sprouting of endothelial cells from preexisting blood vessel. The tumor environment, which counts on the tumor cell proliferation, is plenty of proangiogenic factors, such as angiogenin, TGF (α and ß), FGF, VEGF, all of them playing a crucial role in angiogenesis, an important hallmark of cancer frequently related to a poor prognosis. Therefore, therapies focusing the inhibition of cancer neovasculogenesis have become an interesting strategy for the development of antitumor therapies. In this work, we investigate the effect of tick saliva on the human endothelial cells, in order to understand its inhibitory effects on angiogenesis. To this end, the HUVEC cells were used as model of angiogenesis in vitro and the anti-proliferative, anti-migratory, cytotoxicity was evaluated. Our data depicts that saliva impairs cell development by causing structural changes while precludes cell proliferation and migration, that are crucial events related to angiogenesis. Aiming the identification of the bioactive components related to antiangiogenic activity, saliva was analyzed through the Mass Spectrometry and among all molecules identified, disintegrins and cathepsin L seems to be primarily responsible for the antiangiogenic effects of saliva.

Suppression of Plant Defenses by Herbivorous Mites Is Not Associated with Adaptation to Host Plants.

Some herbivores suppress plant defenses, which may be viewed as a result of the coevolutionary arms race between plants and herbivores. However, this ability is usually studied in a one-herbivore-one-plant system, which hampers comparative studies that could corroborate this hypothesis. Here, we extend this paradigm and ask whether the herbivorous spider-mite Tetranychus evansi, which suppresses the jasmonic-acid pathway in tomato plants, is also able to suppress defenses in other host plants at different phylogenetic distances from tomatoes. We test this using different plants from the Solanales order, namely tomato, jimsonweed, tobacco, and morning glory (three Solanaceae and one Convolvulaceae), and bean plants (Fabales). First, we compare the performance of T. evansi to that of the other two most-commonly found species of the same genus, T. urticae and T. ludeni, on several plants. We found that the performance of T. evansi is higher than that of the other species only on tomato plants. We then showed, by measuring trypsin inhibitor activity and life history traits of conspecific mites on either clean or pre-infested plants, that T. evansi can suppress plant defenses on all plants except tobacco. This study suggests that the suppression of plant defenses may occur on host plants other than those to which herbivores are adapted.

Gene cloning and difference analysis of vitellogenin in Neoseiulus barkeri (Hughes).

Neoseiulus barkeri (HUGHES) is the natural enemy of spider mites, whiteflies and thrips. Screening for chemically-resistant predatory mites is a practical way to balance the contradiction between the pesticide using and biological control. In this study, the number of eggs laid by fenpropathrin-susceptible and resistant strains of N. barkeri was compared. Additionally, we cloned three N. barkeri vitellogenin (Vg) genes and used quantitative real-time polymerase chain reaction to quantify Vg expression in susceptible and resistant strains. The total number of eggs significantly increased in the fenpropathrin-resistant strain. The full-length cDNA cloning of three N. barkeri Vg genes (NbVg1, NbVg2 and NbVg3) revealed that the open reading frames of NbVg1, NbVg2 and NbVg3 were 5571, 5532 and 4728 bp, encoding 1856, 1843 and 1575 amino acids, respectively. The three N. barkeri Vg possessed the Vitellogenin-N domain (or lipoprotein N-terminal domain (LPD_N)), von Willebrand factor type D domain (VWD) and the domain with unknown function 1943 (DUF1943). The NbVg1 and NbVg2 expression levels were significantly higher in the resistant strain than in the susceptible strain, while the NbVg3 expression level was lower in the resistant strain. Thus, we speculate that the increased number of eggs laid by the fenpropathrin-resistant strain of N. barkeri may be a consequence of changes in Vg gene expression.

Complex Evolutionary Dynamics of Massively Expanded Chemosensory Receptor Families in an Extreme Generalist Chelicerate Herbivore.

While mechanisms to detoxify plant produced, anti-herbivore compounds have been associated with plant host use by herbivores, less is known about the role of chemosensory perception in their life histories. This is especially true for generalists, including chelicerate herbivores that evolved herbivory independently from the more studied insect lineages. To shed light on chemosensory perception in a generalist herbivore, we characterized the chemosensory receptors (CRs) of the chelicerate two-spotted spider mite, Tetranychus urticae, an extreme generalist. Strikingly, T. urticae has more CRs than reported in any other arthropod to date. Including pseudogenes, 689 gustatory receptors were identified, as were 136 degenerin/Epithelial Na+ Channels (ENaCs) that have also been implicated as CRs in insects. The genomic distribution of T. urticae gustatory receptors indicates recurring bursts of lineage-specific proliferations, with the extent of receptor clusters reminiscent of those observed in the CR-rich genomes of vertebrates or C. elegans Although pseudogenization of many gustatory receptors within clusters suggests relaxed selection, a subset of receptors is expressed. Consistent with functions as CRs, the genomic distribution and expression of ENaCs in lineage-specific T. urticae expansions mirrors that observed for gustatory receptors. The expansion of ENaCs in T. urticae to > 3-fold that reported in other animals was unexpected, raising the possibility that ENaCs in T. urticae have been co-opted to fulfill a major role performed by unrelated CRs in other animals. More broadly, our findings suggest an elaborate role for chemosensory perception in generalist herbivores that are of key ecological and agricultural importance.

Cloning and differential expression of five heat shock protein genes associated with thermal stress and development in the polyphagous predatory mite Neoseiulus cucumeris (Acari: Phytoseiidae).

In order to explore the role of heat shock proteins (Hsps) during thermal stress and development in the predatory mite Neoseiulus cucumeris (Oudemans), we cloned and characterized five full-length Hsp genes. We investigated the expression levels of these genes by quantitative real-time PCR. The five genes characterized here were NcHsp90, NcHsp75, NcHsp70, NcHsp60, and NcHsp40. These Hsps showed high sequence conservation and had greatest identity with heat shock proteins of Metaseiulus occidentalis and other mite and insect species. All five NcHsp genes showed changes in their levels of expression during development. Higher levels of expression were observed in adult females than in adult males, but there were no significant changes between pre-oviposition and post-oviposition stages in the females. NcHsp90, NcHsp75, and NcHsp70 expression levels were up-regulated after a heat shock, and the increases in NcHsp75 and NcHsp70 expression levels were maintained for at least 3 h. Up-regulation of NcHsp60 and NcHsp40 was not detected after 1 h at a high temperature (35-45 °C); however, a significant down-regulation was observed after 3 h heat exposure at 35 °C and 3 h recovery at 25 °C. Cold shock treatment (-5 to 15 °C) for 1 h did not acute elicit changes in the expression levels of any of the genes. At 5 °C, the expression levels of NcHsp90 significantly increased after 6 or 24 h exposure compared to the levels after 1 h exposure. Thus, expression of Hsp genes in N. cucumeris reflected developmental changes, sexual difference, and variable induced response to thermal stress. Increased expression of Hsps might protect N. cucumeris individuals under extreme temperature conditions. Therefore, it may be possible to enhance the thermal tolerance of commercially available N. cucumeris using temperature acclimation. Treatment at 35 °C should be suitable for such acclimation.

The oviduct is a brood chamber for facultative egg retention in the parthenogenetic oribatid mite Archegozetes longisetosus AOKI (Acari, Oribatida).

Archegozetes longisetosus is a parthenogenetic oribatid mite and a chelicerate model organism. We examined the localisation of processes between vitellogenesis and embryogenesis as well as the anatomy and histology of involved structures by means of light- and electron microscopy. The proximal oviduct is differentiated into an oviductal bulb, exhibiting a strong secretory epithelium. Here, solidification of the egg shell instantaneously occurs upon passing of the egg from the perivitelline space into the oviductal lumen. This is interpreted as an internalised oviposition with the generation boundary being effectively located at the ovary-oviduct transition, rendering the oviducts into functional brood chambers. The parity mode combines elements of oviparity and ovolarviparity with facultative egg retention.

A laboratory assessment of the toxic attributes of six 'reduced risk insecticides' on Galendromus occidentalis (Acari: Phytoseiidae).

The modified excised leaf disc method was used to measure the effects of six insecticides on eggs, larvae, adults, and female fecundity of Galendromus occidentalis (Nesbitt) in a 'worst case laboratory exposure'. This study identified insecticides that would be recommended for tier II field evaluations for an integrated pest management program. Commercially formulated insecticides were applied with a thin-layer chromatography sprayer adjusted to 10.34 kPa (1.5 psi), at the recommended label concentrations in Canada. LC(50) values were estimated from aliquots above and below that concentration. Spinetoram and spirotetramat were toxic at label concentrations. The label concentration for spinetoram was 34.3-fold the LC(50) estimate (0.006 gL(-1)) and for spirotetramat the label concentration was 7.7-fold the LC(50) estimate (0.03 gL(-1)). Clothianidin was considerably less toxic and the label concentration was 0.15-fold the LC(50) estimate (2.29 gL(-1)). Estimates of LC(50) for novaluron and chlorantraniliprole could not be established. Both materials showed slight toxicity to at least one growth stage of the predator. Novaluron, clothianidin and chlorantraniliprole should be evaluated in the field for compatibility in IPM programs. Flubendiamide was harmless to all growth stages and it is recommended for inclusion in IPM programs without additional tier II field evaluations. Field evaluations with spinetoram and spirotetramat should be pursued only if alternatives are unavailable.

Influence of selected fruit tree pollen on life history of Euseius stipulatus (Acari: Phytoseiidae).

Euseius stipulatus (Athias-Henriot) is a predatory mite widespread in the Mediterranean region considered to be important for the biological control of spider mites in citrus orchards. Development, survival and reproduction of this phytoseiid mite feeding on seven commercially obtained pollen were studied under constant laboratory conditions (20 +/- 1 degrees C, RH 65 +/- 5%, photoperiod 16L: 8D h). Mites were kept individually at rearing units with ample quantity of almond (Prunus amygdalus Batch), apple (Malus domestica Borkh), apricot (Prunus armeniaca L.), cherry (Prunus avium L.), pear (Pyrus communis L.), plum (Prunus domestica L.) and walnut (Juglans regia L.) pollen as food source. Developmental time from egg to adult varied between the several pollen tested from 8.38 +/-0.08 to 9.58 +/- 0.11 days for females and from 8.23 +/-0.12 and 9.07 +/-0.12 days for males. Female longevity varied from 11.53 +/- 1.22 to 51.38 +/- 2.45 days, while fecundity ranged from 22.84 +/- 2.30 to 43.61 +/- 3.78 eggs/female. The predator was unable to reproduce when feeding on walnut pollen. Data were submitted to life table analysis and values of the intrinsic rate of increase were derived, ranging from 0.079 to 0.146 (day(-1)). The cumulative Weibull function that was used to describe the age specific survival of females produced excellent fits to the survival data. Results show that almond, plum, cherry and apricot pollen possess higher nutritional value for E. stipulatus than pear and apple pollen and thus may contribute in sustaining and increasing the predator population in field conditions. Walnut pollen can be utilized by the predator only to survive during short periods of time when principal or alternative food sources are scarce.

Acarine regulators of water hyacinth in Kerala (India).

Screening of the mite fauna on water hyacinth, Eichhornia crassipes, at 25 localities throughout Kerala (India) yielded a list of 21 phytophagous species from nine families. Distribution of mites at these localities and type of injury produced suggested that Oligonychus biharensis and Orthogalumna terebrantis warrant further study. In an experimental set-up the injury caused by these mites was quantified. Infestation by Ol. biharensis lead to reduced photosynthesis by the host (significant loss in chlorophyll content). Injurious effects of Or. terebrantis involved leaf mining by the developing instars leading to wilting of the leaves resulting in reduced dry weight. The relative merits of the two species as potential control agents of water hyacinth are discussed.

A possible pathway of chitin synthesis as revealed by electron microscopy in Tetranychus urticae (Acari, Tetranychidae).

Two chitin synthesising systems in Tetranychus urticae area described: one chitosomal system located in the oocytes where spatial and temporal distances are large, and one membrane bound system located in the hypodermis. Similarity of mechanisms of chitin synthesis in animals and plants is suggested.

DNA-content in isolated nuclei of postembryonic stages of progeny from normal and irradiated males of Tetranychus urticae Koch (Acari, Tetranychidae).

The 1C DNA-content of isolated nuclei of postembryonic stages of Tetranychus urticae stained with the classic and a modified Schiff's reagent was cytophotometrically estimated as 0.1 pg, a low value in animals. For many tissues of this arrhenotokous species the ploidy ratio between males and females is 1:2, indicating the absence of sex-related differences in ploidy.--In addition, DNA measurements were performed to evaluate irradiation-experiments, starting with X-irradiation of mature sperm in males with doses known from previous work to induce chromosomal fragments that are subject to loss and missegregation in the embryonic mitotic stages of the female progeny despite the presumed holokinetic nature of the chromosomes. The DNA-content of the nuclei of the surviving postembryonic preadult stages did not indicate the occurrence of nuclei with in-between male/female values, ruling out loss and missegregation of fragments as important factors in postembryonic lethality. Abnormally low DNA-values in some adult females could be attributed to development of embryos before oviposition caused by radiation-induced effects.