Molecular biology and functions of epilysin (MMP-28)

Epilysin (MMP-28) is the most recently identified member of the matrix metalloproteinase (MMP) family of extracellular proteases. Together these enzymes are capable of degrading almost all components of the extracellular matrix (ECM) and are thus involved in important biological processes such as development, wound healing and immune functions, but also in pathological processes such as tumor invasion, metastasis and arthritis. MMPs do not act solely by degrading the ECM. They also regulate cell behavior by releasing growth factors and biologically active peptides from the ECM, by modulating cell surface receptors and adhesion molecules and by regulating the activity of many important mediators in inflammatory pathways. The aim of this study was to define the unique role of epilysin within the MMP-family, to elucidate how and when it is expressed and how its catalytic activity is regulated. To gain information on its essential functions and substrates, the specific aim was to characterize how epilysin affects the phenotype of epithelial cells, where it is biologically expressed. During the course of the study we found that the epilysin promoter contains a well conserved GT-box that is essential for the basic expression of this gene. Transcription factors Sp1 and Sp3 bind this sequence and could hence regulate both the basic and cell type and differentiation stage specific expression of epilysin. We cloned mouse epilysin cDNA and found that epilysin is well conserved between human and mouse genomes and that epilysin is glycosylated and activated by furin. Similarly to in human tissues, epilysin is normally expressed in a number of mouse tissues. The expression pattern differs from most other MMPs, which are expressed only in response to injury or inflammation and in pathological processes like cancer. These findings implicate that epilysin could be involved in tissue homeostasis, perhaps fine-tuning the phenotype of epithelial cells according to signals from the ECM. In view of these results, it was unexpected to find that epilysin can induce a stable epithelial to mesenchymal transition (EMT) when overexpressed in epithelial lung carcinoma cells. Transforming growth factor b (TGF-b) was recognized as a crucial mediator of this process, which was characterized by the loss of E-cadherin mediated cell-cell adhesion, elevated expression of gelatinase B and MT1-MMP and increased cell migration and invasion into collagen I gels. We also observed that epilysin is bound to the surface of epithelial cells and that this interaction is lost upon cell transformation and is susceptible to degradation by membrane type-1-MMP (MT1-MMP). The wide expression of epilysin under physiological conditions implicates that its effects on epithelial cell phenotype in vivo are not as dramatic as seen in our in vitro cell system. Nevertheless, current results indicate a possible interaction between epilysin and TGF-b also under physiological circumstances, where epilysin activity may not induce EMT but, instead, trigger less permanent changes in TGF-b signaling and cell motility. Epilysin may thus play an important role in TGF-b regulated events such as wound healing and inflammation, processes where involvement of epilysin has been indicated.

Comparative studies on the invasion of cattle ticks (Rhipicephalus (Boophilus) microplus) and sheep blowflies (Lucilia cuprina) by Metarhizium anisopliae (Sorokin).

Microscopic investigations over time were carried out to study and compare the pathogenesis of invasion of ticks and blowflies by Metarhizium anisopliae. The scanning electron microscope and stereo light microscope were used to observe and record processes on the arthropods' surfaces and the compound light microscope was used to observe and record processes within the body cavities. Two distinctly different patterns of invasion were found in ticks and blowflies. Fungal conidia germinated on the surface of ticks then hyphae simultaneously penetrated into the tick body and grew across the tick surface. There was extensive fungal degradation of the tick cuticle, particularly the outer endocuticle. Although large numbers of conidia adhered to the surface of blowflies, no conidia were seen to germinate on external surfaces. A single germinating conidium was seen in the entrance to the buccal cavity. Investigations of the fly interior revealed a higher density of hyphal bodies in the haemolymph surrounding the buccal cavity than in haemolymph from regions of the upper thorax. This pattern suggests that fungal invasion of the blowfly is primarily through the buccal cavity. Plentiful extracellular mucilage was seen around the hyphae on tick cuticles, and crystals of calcium oxalate were seen amongst the hyphae on the surface of ticks and in the haemolymph of blowflies killed by M. anisopliae isolate ARIM16.

Introduction, establishment, and potential geographic range of Carmenta sp. nr ithacae (Lepidoptera: Sesiidae), a biological control agent for Parthenium hysterophorus (Asteraceae) in Australia

Parthenium (Parthenium hysterophorus L.), a major weed causing economic, environmental, and human and animal health problems in Australia and several countries in Asia, Africa, and the Pacific, has been a target for biological control in Australia since the mid-1970s. Nine species of insects and two rust fungi have been introduced as biological control agents into Australia. These include Carmenta sp. nr ithacae, a root feeding agent from Mexico. The larvae of C. sp. nr ithacae bore through the stem-base into the root where they feed on the cortical tissue of the taproot. During 1998-2002, 2,816 larval-infested plants and 387 adults were released at 31 sites across Queensland, Australia. Evidence of field establishment was first observed in two of the release sites in central Queensland in 2004. Annual surveys at these sites and nonrelease sites during 2006-2011 showed wide variations in the incidence and abundance of C. sp. nr ithacae between years and sites. Surveys at three of the nine release sites in northern Queensland and 16 of the 22 release sites in central Queensland confirmed the field establishment of C. sp. nr ithacae in four release sites and four nonrelease sites, all in central Queensland. No field establishment was evident in the inland region or in northern Queensland. A CLIMEX model based on the native range distribution of C. sp. nr ithacae predicts that areas east of the dividing range along the coast are more suitable for field establishment than inland areas. Future efforts to redistribute this agent should be restricted to areas identified as climatically favorable by the CLIMEX model.

Complementing biological control with plant suppression: Implications for improved management of parthenium weed (Parthenium hysterophorus L.)

Parthenium hysterophorus L. is a weed of global significance that has become a major weed in Australia and many other parts of the world. A combined approach for the management of parthenium weed using biological control and plant suppression, was tested under field conditions over a two-year period in southern central Queensland. The six suppressive plant species, selected for their demonstrably suppressive ability in earlier glasshouse studies, worked synergistically with the biological control agents (Epiblema strenuana Walker, Zygogramma bicolorata Pallister, Listronotus setosipennis Hustache and Puccinia abrupta var. partheniicola) present in the field to reduce the growth (above ground biomass) of parthenium weed, by between 60–86% and 47–91%, in Years 1 and 2, respectively. The biomass of the suppressive plants was between 6% and 23% greater when biological control agents were present than when the biological control agents had been excluded. This shows that parthenium weed can be more effectively managed by combining the current biological control management strategy with selected sown suppressive plant species, both in Australia and elsewhere.

Cat's claw creeper leaf-mining jewel beetle Hylaeogena jureceki Obenberger (Coleoptera: Buprestidae), a host-specific biological control agent for Dolichandra unguis-cati (Bignoniaceae) in Australia

Cat's claw creeper, Dolichandra unguis-cati (L.) L.G. Lohman (syn: Macfadyena unguis-cati (L.) A.H. Gentry) (Bignoniaceae), a major environmental weed in Queensland and New South Wales, is a Weed of National Significance and an approved target for biological control. A leaf-mining jewel beetle, Hylaeogena jureceki Obenberger (Coleoptera: Buprestidae), first collected in 2002 from D. unguis-cati in Brazil and Argentina, was imported from South Africa into a quarantine facility in Brisbane in 2009 for host-specificity testing. H. jureceki adults chew holes in leaves and lay eggs on leaf margins and the emerging larvae mine within the leaves of D. unguis-cati. The generation time (egg to adult) of H. jureceki under quarantine conditions was 55.4 ± 0.2 days. Host-specificity trials conducted in Australia on 38 plant species from 11 families supplement and support South African studies which indicated that H. jureceki is highly host-specific and does not pose a risk to any non-target plant species in Australia. In no-choice tests, adults survived significantly longer (>32 weeks) on D. unguis-cati than on non-target test plant species (<3 weeks). Oviposition occurred on D. unguis-cati and one non-target test plant species, Citharexylum spinosum (Verbenaceae), but no larval development occurred on the latter species. In choice tests involving D. unguis-cati, C. spinosum and Avicennia marina (Avicenniaceae), feeding and oviposition were evident only on D. unguis-cati. The insect was approved for field release in Australia in May 2012.

Cat's claw creeper vine, Macfadyena unguis-cati (Bignoniaceae), invasion impacts: comparative leaf nutrient content and effects on soil physicochemical properties

Macfadyena unguis-cati (L.) Gentry (Bignoniaceae) is a major environmental weed in coastal Queensland, Australia. There is a lack of quantitative data on its leaf chemistry and its impact on soil properties. Soils from infested vs uninfested areas, and leaves of M. unguis-cati and three co-occurring vine species (one exotic, two native) were collected at six sites (riparian and non-riparian) in south-eastern Queensland. Effects of invasion status, species, site and habitat type were examined using univariate and multivariate analyses. Habitat type had a greater effect on soil nutrients than on leaf chemistry. Invasion effect of M. unguis-cati on soil chemistry was more pronounced in non-riparian than in riparian habitat. Significantly higher values were obtained in M. unguis-cati infested (vs. uninfested) soils for ~50% of traits. Leaf ion concentrations differed significantly between exotic and native vines. Observed higher leaf-nutrient load (especially nitrogen, phosphorus and potassium) in exotic plants aligns with the preference of invasive plant species for disturbed habitats with higher nutrient input. Higher load of trace elements (aluminium, boron, cadmium and iron) in its leaves suggests that cycling of heavy-metal ions, many of which are potentially toxic at excess level, could be accelerated in soils of M. unguis-cati-invaded landscape. Although inferences from the present study are based on correlative data, the consistency of the patterns across many sites suggests that M. unguis-cati may improve soil fertility and influence nutrient cycling, perhaps through legacy effects of its own litter input.

Impact of a biological control agent, Chiasmia assimilis, on prickly acacia (Acacia nilotica ssp. indica) seedlings

A leaf-feeding geometrid, Chiasmia assimilis (Warren), was introduced into northern Queensland from South Africa in 2002 as a biological control agent for the invasive woody weed, prickly acacia, Acacia nilotica subsp. indica (Bentham) Brenan. The insect established in infestations in coastal areas between the townships of Ayr and Bowen where the larvae periodically cause extensive defoliation at some localities during summer and autumn. The impact of this herbivory on a number of plant parameters, including shoot length, basal stem diameter, root length, number of leaves, number of branches, and above and below ground biomass was investigated at one coastal site through an insect exclusion trial using potted seedlings and regular spray applications of a systemic insecticide to exclude the biological control agent. Half the seedlings, both sprayed and unsprayed, were placed beneath the prickly acacia canopy, the other half were placed in full sunlight. Larvae of C. assimilis were found on unsprayed seedlings in both situations. The effects of herbivory, however, were significant only for seedlings grown beneath the canopy. At the end of the five-month trial period, shoot length of these seedlings was reduced by 30%, basal stem diameter by 44%, root length by 15%, number of leaves by 97%, above ground biomass by 87%, and below ground biomass by 77% when compared to sprayed seedlings. Implications are that the insect, where established, may reduce seedling growth beneath existing canopies and in turn may help limit the formation of dense infestations. Crown Copyright (C) 2012 Published by Elsevier Inc. All rights reserved.

Improving decisions for invasive species management: Reformulation and extensions of the Panetta-Lawes eradication graph

Aim: Effective decisions for managing invasive species depend on feedback about the progress of eradication efforts. Panetta & Lawes. developed the eradograph, an intuitive graphical tool that summarizes the temporal trajectories of delimitation and extirpation to support decision-making. We correct and extend the tool, which was affected by incompatibilities in the units used to measure these features that made the axes impossible to interpret biologically. Location: Victoria, New South Wales and Queensland, Australia. Methods: Panetta and Lawes' approach represented delimitation with estimates of the changes in the area known to be infested and extirpation with changes in the mean time since the last detection. We retain the original structure but propose different metrics that improve biological interpretability. We illustrate the methods with a hypothetical example and real examples of invasion and treatment of branched broomrape (Orobanche ramosa L.) and the guava rust complex (Puccinia psidii (Winter 1884)) in Australia. Results: These examples illustrate the potential of the tool to guide decisions about the effectiveness of search and control activities. Main conclusions: The eradograph is a graphical data summary tool that provides insight into the progress of eradication. Our correction and extension of the tool make it easier to interpret and provide managers with better decision support. © 2013 John Wiley & Sons Ltd.

Biological Control of Oilseed Rape Pests with Entomopathogenic Nematodes

Biological control techniques attract increasing attention as one of the sustainable alternatives to pesticide use in integrated pest management programs. In order to develop sustainable pest management methods for arable crops based on entomopathogenic nematodes (EPN), their efficacy and persistence needed to be investigated, and an economically feasible delivery system had to be developed. In this study, first a survey of entomopathogens was conducted, and a system approach was tested, using the oilseed Brassica (OSB) growing system (OSB, spring wheat, and red clover) as a model. The system approach aimed at determining the potential of Steinernema feltiae (Filipjev) for the control of OSB pests, developing OSB rotation schemes that support EPN persistence, and investigating the impact of the selected biotic and abiotic factors on efficacy and persistence of EPN. This study employed abductive logic (which employs constant interplay between the theory and empirical observation), quantitative methods, and a case study on OSB. Laboratory and field experiments were carried out, and two types of pathogen surveys. A horizontal survey included OSB fields across Estonia, Germany, Poland, Sweden and the UK, while a vertical survey included sampling from two sets of differently managed experimental fields during three years. A new approach was introduced for measuring occurrence, where the prevalence and relative intensity of entomopathogens, biotic agents, and unidentified insect antagonists were determined. The effect of dose, timing, and the application method on S. feltiae in the control of pests in OSB, and the potential of a controlled release delivery system (CRS) were evaluated in the field. Studies on the impact of selected biotic and abiotc factors (Brassica plant, bait insects, developmental stages of Meligethes aeneus Fab., Isaria fumosorosea Wize (Ifr), and organic and synthetic fertilizers) on the efficacy of S. feltiae were conducted in the laboratory. Persistence of S. feltiae in the OSB growing system, and the effect of dose, timing, and the application method, was assessed in the field as part of the efficacy experiments. The impact of selected biotic and abiotic factors on S. feltiae persistence was assessed in laboratory experiments. The pathogen survey showed that the occurrence of entomopathogens is low in the OSB growing system, and that a management system causing less disturbance (ICM) to the soil increases the relative intensity of insect parasitic nematodes and other insect antagonists. A longer study period is required to show any possible impact of ICM on the relative intensity of entomopathogenic fungi, or on the prevalence of entomopathogens. Two different measures of the occurrence yielded different results: the relative intensity revealed the difference between the two different crop management methods, while prevalence did not. The highest efficacy of S. feltiae was achieved by using a low dose and targeting all stages of M. aeneus. When only the larval stage was targeted, the application method and dose had no significant effect. The CRS decreased the pest abundance significantly more than the surface application method. S. feltiae persisted in the OSB fields in Finland for several months, but did not survive the winter. The strain survived for 7 months when it was applied in autumn in Germany, but its populations declined rapidly after winter. The examined biotic and abiotic factors had variable impacts on S. feltiae efficacy and persistence. The two measures, prevalence and relative intensity of entomopathogens, gave valuable information for their use in biocontrol programs. The recommended biocontrol strategy for OSB growing in Finland is inundation and seasonal inoculation of EPN. The impact of some biotic and abiotic factors on S. feltiae efficacy and persistence is significant, and can be used to improve the efficacy of EPN. The CRS is a novel alternative for EPN application, and should also be considered for use on other crops. Keywords: Biological control, inundation, inoculation, conservation, formulation, slow release method, crop rotation, Entomopathogenic nematodes, Steinernema feltiae, oilseed rape pests, Meligethes aeneus, Phyllotreta spp., occurrence, prevalence, intensity, efficacy, persistence, field, Isaria fumosorosea, biotic factors, abiotic factors, interaction, impact, insect stages, integrated crop management, standard (conventional) crop management

Development of cat's claw creeper leaf-tying moth Hypocosmia pyrochroma (Lepidoptera: Pyralidae) at different temperatures: Implications for establishment as a biological control agent in Australia and South Africa

The leaf-tying moth Hypocosmia pyrochroma Jones (Lepidoptera: Pyralidae), a native of sub tropical South America, has been introduced as a biological control agent for cat’s claw creeper, Dolichandra unguis-cati (L.) Lohman (Bignoniaceae), in Australia and South Africa. So far there has been no evidence of its field establishment in either country. A narrow temperature tolerance is a potential limiting factor for the establishment of weed biological control insects in novel habitats. In this study, we evaluated the effect of seven constant temperatures (12–40 °C) on the survival and development of H. pyrochroma in temperature-controlled cabinets. Temperatures between 20 and 30 °C were the most favorable for adult survival, oviposition, egg hatching, and larval and pupal development. Adult survival (12–40 °C) and egg development (15–35 °C) showed tolerance for wider temperature ranges than oviposition, and larval and pupal development, which were all negatively affected by both high (>30 °C) and low (<20 °C) temperatures. The degree-day (DD) requirement to complete a generation was estimated as 877 above a threshold temperature of 12 °C. Based on DD requirements and an obligatory winter diapause of pupae from mid-autumn to mid-spring, the potential number of generations (egg to adult) the leaf-tying moth can complete in a year in Australia or South Africa range from one to three. A climate-matching model predicted that the inland regions of both Australia and South Africa are less favorable for H. pyrochroma than the coastal areas. The study suggested that H. pyrochroma is more likely to establish in the coastal areas of Australia where most of the cat’s claw creeper infestations occur, than in South Africa where most of the cat’s claw creeper infestations are inland.