Development of a cDNA microarray for the measurement of gene expression in the sheep scab mite Psoroptes ovis

Background: Sheep scab is caused by the ectoparasitic mite Psoroptes ovis which initiates a profound cutaneous inflammatory response, leading to the development of the skin lesions which are characteristic of the disease. Existing control strategies rely upon injectable endectocides and acaricidal dips but concerns over residues, eco-toxicity and the development of acaricide resistance limit the sustainability of this approach. In order to identify alternative means of disease control, a deeper understanding of both the parasite and its interaction with the host are required. Methods: Herein we describe the development and utilisation of an annotated P. ovis cDNA microarray containing 3,456 elements for the measurement of gene expression in this economically important ectoparasite. The array consists of 981 P. ovis EST sequences printed in triplicate along with 513 control elements. Array performance was validated through the analysis of gene expression differences between fed and starved P. ovis mites. Results: Sequences represented on the array include homologues of major house dust mite allergens and tick salivary proteins, along with factors potentially involved in mite reproduction and xenobiotic metabolism. In order to validate the performance of this unique resource under biological conditions we used the array to analyse gene expression differences between fed and starved P. ovis mites. These analyses identified a number of house dust mite allergen homologues up-regulated in fed mites and P. ovis transcripts involved in stress responses, autophagy and chemosensory perception up-regulated in starved mites. Conclusion: The P. ovis cDNA microarray described here has been shown to be both robust and reproducible and will enable future studies to analyse gene expression in this important ectoparasite

Gene silencing by RNA interference in the ectoparasitic mite, Psoroptes ovis

Abstract The presence of components of the RNA interference (RNAi) pathway in Psoroptes ovis, an ectoparasitic mite responsible for psoroptic mange, was investigated through interrogation of the P. ovis genome. Homologues of transcripts representing critical elements for achieving effective RNAi in the mite, Tetranychus urticae and the model organisms Caenorhabditis elegans and Drosophila melanogaster were identified and, following the development of a non-invasive immersion method of double stranded RNA delivery, gene silencing by RNAi was successfully demonstrated in P. ovis. Significant reductions in transcript levels were achieved for three target genes which encode the Group 2 allergen (Pso o 2), mu-class glutathione S-transferase (PoGST-mu1) and beta-tubulin (Poβtub). This is the first demonstration of RNAi in P. ovis and provides a mechanism for mining transcriptomic and genomic datasets for novel control targets against this economically important ectoparasite

Revisiting probabilistic seismic hazard analysis of Gujarat: an assessment of Indian design spectra

Seismic hazard assessment is the key tool for rational planning, safety and design of infrastructures in seismically vulnerable regions. Gujarat is the only state in peninsular India with the maximum seismic hazard of large shallow earthquakes originating from intra-plate seismicity. Probabilistic seismic hazard assessment (PSHA) of Gujarat is carried out in this paper. Three seismogenic sources, namely Kutch, Saurashtra and Mainland Gujarat, are considered, and seismicity parameters are estimated separately for each region taking into account the completeness of the available earthquake data. Peak ground acceleration (PGA) of the horizontal component and spectral acceleration at specific periods are considered as the intensity measures. Ground motion predictive equation chosen was reported to be based on simulated ground motions and verified against the strong motion records in the study region. Results are reported for the 17 major cities at the bedrock and also for the soil sites. Apart from hazard curves, 2475 and 475 years of return periods are considered for the PGA and uniform hazard spectra (UHS). The results are compared with the present recommendations of Indian Standards. Key observations include (1) Indian Standards underpredict PGA in the entire Gujarat when the soil sites are considered and in a few cities even at the bedrock; (2) amplification of PGA (or short period hazard) on account of soil sites should be included in the Indian Standard, which is currently absent; (3) shape of the UHS indicates that a separate amplification is required at the hyperbolic portion; and (4) ratio of 2475–475 years of PGA, which is considered 2.0 in Indian Standard, should be reduced to 1.5. Time-dependent recurrence model is also included in this paper and compared with conventional PSHA. General observations include that (1) hazard may increase significantly on account of time dependency; (2) this also influences the disaggregation and in turn the selection of ground motions; and (3) time since last earthquake significantly influences the extent of the effect of time dependency.by Asim Bashir and Dhiman Bas