Proteinase activity in the midgut of the pentatomid stinkbug Podisus maculiventris was investigated. The optimal pH for adult and nymph proteolysis was pH 6.0 and pH 6.5, respectively. Proteinase activity was characterised using a range of diagnostic inhibitors. Activity of both adult and nymphal gut extracts, detected by the hydrolysis of Z-Phe-Arg-pNA, was inhibited to <20% of control levels by several inhibitors (e.g. E-64 and chicken egg white cystatin) associated with the inhibition of cysteine proteinases. The less specific inhibitor leupeptin reduced proteolytic activity to around 1.0% of the control values. In-gel analysis of the enzymes revealed that proteolytic activity was due to at least four proteinases, of ca. 30, 36, 50 and 110 kDa, which were all susceptible to E-64 inhibition. Salivary gland extracts gave maximal activity at pH 8.0 when tested for general proteolytic activity using fluorescent BODIPY-FL casein substrate, and showed moderate levels of inhibition when incubated with inhibitors of serine-, cysteine-, aspartic- and metallo-proteinases. Leupeptin and PMSF gave the highest levels of inhibition of salivary proteolytic activity, at ca. 50%, whilst the plant-derived inhibitors SKTI, CpTI and OC-1 did not inhibit proteolysis.
Ixodid ticks remain attached to their hosts for several days to weeks. During this extended feeding process new proteins involved in the modulation of host immune responses are expressed in tick salivary glands. In our study a stimulatory or inhibitory effect of salivary gland extracts (SGE) of unfed and partially fed female Ixodes ricinus (Linnaeus, 1758), female and male Amblyomma variegatum (Fabricius, 1794) and Rhipicephalus appendiculatus Neumann, 1901 ticks on human lymphocyte proliferation induced by Concanavalin A (ConA) and phytohaemagglutinin (PHA), respectively, was investigated. SGE of all female ticks examined suppressed proliferation of ConA-induced lymphocytes; highly significant suppression was observed in the presence of unfed I. ricinus and 9-day fed A. variegatum SGE. SGE of partially fed A. variegatum and I. ricinus females suppressed PHA responses of lymphocytes. Lymphocytes showed reduced PHA and ConA responses in the presence of SGE of unfed and 2-day fed R. appendiculatus females, while SGE of 6-day fed females enhanced PHA responses, but reduced their ConA responses; generally SGE of 2-day fed females displayed the strongest inhibition. Amblyomma variegatum male SGE slightly enhanced PHA, but significantly reduced ConA responses of lymphocytes and their inhibitory effect increased during feeding. SGE of unfed and 2-day fed R. appendiculatus males enhanced PHA and ConA responses and those of 6-day fed males suppressed lymphocyte proliferation. The results suggest that (i) species- and sex-specific differences exist in the effects of tick salivary gland antigens on human lymphocyte proliferation and (ii) effect of SGE on human lymphocyte responses to mitogens varies depending on the tick feeding status.
The morphology of the male salivary glands of eighteen species of Panorpidae from China was studied using light microscopy. The results show that the male salivary glands differ markedly both at generic and specific levels. In Neopanorpa, the salivary glands consist of only two simple long secretory tubes extending to the fifth or sixth abdominal segment, whereas in Sinopanorpa, the salivary glands are composed of six extremely elongated secretory tubes. In Panorpa, the salivary glands are quite diverse, comprising two simple short secretory tubes only extending to the prothorax in the P. amurensis group (P. liui and P. jilinensis), six long tubes in the P. centralis group, eight to twelve in the P. diceras group and of a very variable number in the P. davidi group (especially in P. bifasciata and P. subambra). Morphology of the male salivary glands should be included in future studies on the systematics and phylogeny of the Panorpidae. and Na Ma, Shu-Yu Liu, Bao-Zhen Hua.
Lygus lineolaris (Palisot de Beauvois, 1818) (tarnished plant bug) is a serious pest of cotton (Gossypium hirsutum L.) in the Delta region as compared to cotton in the Hills region of the state of Mississippi in USA. The reason for this is unclear but it was hypothesized that the plant cell wall degrading polygalacturonase enzyme system in the salivary glands of L. lineolaris from the Delta could be better adapted for cotton, which is grown more predominantly in the Delta region than in the Hills region. Expression analysis of three primary polygalacturonase genes (LlPG1, LlPG2 and LlPG3) was conducted in laboratory reared and field collected populations of L. lineolaris. Assay of polygalacturonase enzyme activity was also conducted to compare wild collected populations. Initial laboratory and field data revealed gene expression differences in sex, age, region, and host plant which guided the direction of our subsequent study during 2013 and 2014. Based on the results of this study, we propose that the three genes studied may not be reflective of the entire polygalacturonase enzyme system and may not be solely responsible for the observed adaptation of L. lineolaris to cotton in the Delta region than in the Hills region. Analyses also revealed that the expression of the three targeted polygalacturonase genes was affected by the host plant from which the insects were collected and that adults had higher polygalacturonase expression than nymphs. Taken together, our results provide strong evidence for developmental stage specific and host plant based change in expression of PG genes in the salivary glands of L. lineolaris. This, however, was not reflected in total polygalacturonase enzyme activity which was not significantly different between regions, hosts, sex, or developmental stage., Daniel Fleming, Natraj Krishnan, Fred Musser., and Obsahuje bibliografii