RNA interference in a cestode reveals specific silencing of selected highly expressed gene transcripts

Evolving RNA interference (RNAi) platforms are providing opportunities to probe gene function in parasitic helminths using reverse genetics. Although relatively robust methods for the application of RNAi in parasitic flatworms have been established, reports of successful RNAi are confined to three genera and there are no known reports of the application of RNAi to the class Cestoda. Here we report the successful application of RNAi to a cestode. Our target species was the common ruminant tapeworm, Moniezia expansa which can significantly impact the health/productivity of cattle, sheep and goats. Initial efforts aimed to silence the neuronally expressed neuropeptide F gene (Me-npf-1), which encodes one of the most abundant neuropeptides in flatworms and a homologue of vertebrate neuropeptide Y (NPY). Double stranded (ds)RNAs, delivered by electroporation and soaking (4-8 h), failed to trigger consistent Me-npf-1 transcript knock-down in adult worms; small interfering RNAs (siRNAs) were also ineffective. Identical approaches resulted in significant and consistent transcript knock-down of actin transcript (71 +/- 4%) following soaking in Me-act-1 dsRNA. Similar successes were seen with hydrophobic lipid-binding protein (Me-lbp-1), with a dsRNA inducing significant target transcript reduction (72 +/- 5%). To confirm the validity of the observed transcript knock-downs we further investigated Me-act-1 RNAi worms for associated changes in protein levels, morphology and phenotype. Me-act-1 RNAi worms displayed significant reductions in both filamentous actin immunostaining (62 +/- 3%) and the amount of actin detected in Western blots (54 +/- 13%). Morphologically, Me-act-1 RNAi worms displayed profound tegumental disruption/blebbing. Further, muscle tension recordings from Me-act-1 RNAi worms revealed a significant reduction in both the number of worms contracting in response to praziquantel (20 +/- 12%) and in their contractile ability. These data demonstrate, to our knowledge for the first time, a functional RNAi pathway in a cestode and show that the robust knock-down of abundant gene transcripts is achievable using long dsRNAs following short exposure times. (C) 2009 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

‘It ain’t easy being green’: Sustainable Development between Environment and Economy in Northern Ireland’

Given the relative lack of research on sustainable development in Northern Ireland, this paper focuses on the tensions between environmental governance and regulation on the one hand, and the ‘post-conflict’ imperative for Northern Ireland to compete and grow as a regional economy without continued British state subvention and subsidisation. The paper outlines how this ‘trade-off’ between ‘environment’ and ‘economy’ is essentially misplaced. It argues that this trade-off can be avoided if there is a shift in focus from an ‘environment versus the economy’ policy position to one in which the ‘triple bottom line’ (social, economic and environmental) of sustainable development becomes the over-arching policy agenda. Sustainable development, unlike either orthodox environmental or economic policy, also connects centrally with the unique ‘post-conflict transformation’ agenda of Northern Ireland. For example, promoting a human rights civic culture, tackling socioeconomic inequality and social exclusion, and building a shared future based on supporting sustainable communities and an innovative model of a ‘green(ing) economy’ goes beyond orthodox economic growth. However, it is clear from the Executive’s Programme for Government, failure to support the creation of an independent Environment Protection Agency, and above all the prioritisation of orthodox economic growth based on foreign direct investment that neither environmental protection nor sustainable development is or will be high on the political or policy agenda in Northern Ireland.