Gene amplification and insecticide resistance

Pesticide resistance in arthropods has been shown to evolve by two main mechanisms, the enhanced production of metabolic enzymes, which bind to and/or detoxify the pesticide, and mutation of the target protein, which makes it less sensitive to the pesticide. One route that leads to enhanced metabolism is the duplication or amplification of the structural gene(s) encoding the detoxifying enzyme, and this has now been described for the three main families (esterases, glutathione S-transferases and cytochrome P450 monooxygenases) implicated in resistance. More recently, a direct or indirect role for gene duplication or amplification has been described for target-site resistance in several arthropod species. This mini-review summarises the involvement of gene duplication/amplification in the insecticide/acaricide resistance of insect and mite pests and highlights recent developments in this area in relation to P450-mediated and target-site resistance.     

Applying evidence-based practice in conservation management: Lessons from the first systematic review and dissemination projects

Lack of systematic evaluation of the effectiveness of conservation practices has hindered advances in scientific management of biodiversity. Development of an evidence-based framework of the kind used in the health services has been advocated as an approach to address this problem. Here we report on and evaluate the first two ecological systematic reviews undertaken using this framework. The subjects were the effectiveness of burning as a conservation intervention for UK sub-montane, dry dwarf shrub heaths (heath review) and blanket bog (bog review). Systematic search yielded 13 data sets from seven ‘heath’ articles and 11 data sets from eight ‘bog’ articles. Data from the heath review were of sufficient quality to enable meta-analysis whereas data from the bog review was synthesised by “vote counting”. Meta-analysis of the heath data indicates that floristic diversity is variable, particularly in early post-fire successional sequences, but older stands experienced significantly greater loss of diversity than younger stands. The bog review indicates that there are no consistent changes in floristic composition in response to burning. The evidence-base for upland management by burning is insufficient to generate robust management recommendations highlighting the necessity for further work. Feedback from the dissemination of the reviews provided valuable lessons for future reviews in terms of stakeholder involvement in question formulation and the balance between a reductionist and holistic approach. Systematic reviews are appropriate tools for conservation management and formalise the information available based on the weight of evidence. Evidence will commonly be lacking in many areas of conservation biology and the evidence-based framework effectively exposes this situation in a way that can determine requirements for needs-led research.     

Expectations for population growth at new breeding locations for the vulnerable New Zealand sea lion (Phocarctos hookeri) using a simulation model

Management plans for threatened or recovering large vertebrate species that are increasing in population size and range focus on the establishment of viable populations within set temporal limits. New Zealand (Hookers) sea lions (Phocarctos hookeri) were declared a threatened species in 1997, and New Zealand legislation requires that threatened species of marine mammals must be managed to reduce human-induced mortality and achieve a non-threatened status within 20 years. The present breeding distribution of P. hookeri is highly localised, with over 95% of total annual pup production located at Auckland Islands and almost all of the remainder at Campbell Island. Breeding elsewhere has been ephemeral or restricted to <10 adult females. The only recorded sustainable breeding at a new location has been at Otago, South Island, New Zealand. This breeding population consisted of a total of four breeding females in 2002 and is derived from one immigrant female that gave birth to her first pup in the 1993/1994 breeding season. The New Zealand Department of Conservation management plan specifies that to achieve a non-threatened status P. hookeri (1) at Otago must increase in the number of breeding females to ?10, and (2) must establish ?two new breeding locations within the 20-year time frame, each with ?10 breeding females. This study 1) projects the population growth trends at a new location (Otago) to see if it will achieve ?10 breeding females within the legislated time frame, and (2) examines the likelihood that other breeding locations will establish elsewhere given the demographic information available for this species. We present 20 deterministic and three stochastic Leslie matrix model scenarios for female population growth for the initial years following the start of breeding at a new location. Our results indicate that (1) a new breeding population derived from one immigrant female is unlikely to reach 10 breeding females in 20 years; this duration is more likely to be 23-41 years (deterministic models) or 23-26 years (stochastic model), (2) the likelihood of two new sites establishing within 20 years is unquantifiable, but the probability is low, and (3) if the legislated outcome and time limit are not revised in the population management plan, the feasibility and effectiveness of re-locating young females could be investigated.     

Tylosin detection in animal feed by liquid chromatography-tandem mass spectrometry with enzymatic hydrolysis of the tylosin urea adduct

When the use of tylosin as a feed additive was forbidden by Council Regulation 2821/98, the necessity of a chemical confirmation method for the monitoring of the ban was created. Recently a method was developed for the detection of tylosin in animal feed by means of LC-MS/MS. During the validation high deviating values for the decision limit, detection capability, and repeatability for tylosin in cattle feed were observed, and the presence of urea and the formation of a tylosin urea adduct (TUA) were suggested as possible explanations. In this study two hydrolysis approaches for the TUA adduct were compared, namely, a chemical hydrolysis and an enzymatic hydrolysis with urease. The latter yielded a more complete hydrolysis of urea and was used for further validation. The recovery increased by approximately 15-25% depending on the amount of urea present in the feed (0.5-2%). The decision limit and detection capability were hardly influenced by the enzymatic hydrolysis.     

Detecting interaction(s) and assessing the impact of component subsets in a chemical mixture using fixed-ratio mixture ray designs

An important environmental and regulatory issue is the protection of human health from potential adverse effects of cumulative exposure to multiple chemicals. Earlier literature suggested restricting inference to specific fixed-ratio rays of interest. Based on appropriate definitions of additivity, single chemical data are used to predict the relationship among the chemicals under the zero-interaction case. Parametric comparisons between the additivity model and the model fit along the fixed-ratio ray(s) are used to detect departure from additivity. Collection of data along reduced fixed-ratio rays, where subsets of chemicals of interest are removed from the mixture and the remaining compounds are at the same relative ratios as considered in the full ray, allow researchers to make inference about the effect of the removed chemicals. Methods for fitting simultaneous confidence bands about the difference between the best fitting model and the model predicted under additivity are developed to identify regions along the rays where significant interactions occur. This general approach is termed the “single chemicals required” (SCR) method of analysis. A second approach, termed “single chemicals not required” (SCNR) method of analysis, is based on underlying assumptions about the parameterization of the response surface. Under general assumptions, polynomial terms for models fit along fixed-ratio rays are associated with interaction terms. Consideration is given to the case where only data along the mixture rays are available. Tests of hypotheses, which consider interactions due to subsets of chemicals, are also developed.     

Statistical analysis of interactive cytotoxicity in human epidermal keratinocytes following exposure to a mixture of four metals

Exposure to mixtures of chemicals is an important and relevant environmental issue. Of particular interest is the detection and characterization of departure of biological effects from additivity. Methodology based on the assumption of additivity is used in fitting single-chemicaldata. Interactionsare determined and characterized by making comparisons between the observed and predicted responses at mixtures along a fixed ratio ray of the component substances. Two simultaneous tests are developed for testing for any departure from additivity. Multiple comparisons procedures are used to compare observed responses to that predicted under additivity. A simultaneous confidence band on the predicted responses along the mixture ray is also developed. The methods are illustrated with cytotoxicity data that arise when human epidermal keratinocytes are exposed to a mixture of arsenic, chromium, cadmium, and lead. Synergistic, antagonistic, and additive cytotoxicities were observed at different dose levels of the four-metal mixture.     

Eight urgent,fundamental and simultaneous steps needed to restore ocean health,and the consequences for humanity and the planet of inaction or delay

1. The ocean crisis is urgent and central to human wellbeing and life on Earth; past and current activities are damaging the planet's main life support system for future generations. We are witnessing an increase in ocean heat, disturbance, acidification, bio‐invasions and nutrients, and reducing oxygen levels. Several of these act like ratchets: once detrimental or negative changes have occurred, they may lock in place and may not be reversible, especially at gross ecological and ocean process scales.
2. Each change may represent a loss to humanity of resources, ecosystem function, oxygen production and species. The longer we pursue unsuitable actions, the more we close the path to recovery and better ocean health and greater benefits for humanity in the future.
3. We stand at a critical juncture and have identified eight priority issues that need to be addressed in unison to help avert a potential ecological disaster in the global ocean. They form a purposely ambitious agenda for global governance and are aimed at informing decision‐makers at a high level. They should also be of interest to the general public.
4. Of all the themes, the highest priority is to rigorously address global warming and limit surface temperature rise to 1.5°C by 2100, as warming is the pre‐eminent factor driving change in the ocean. The other themes are establishing a robust and comprehensive High Seas Treaty, enforcing existing standards for Marine Protected Areas and expanding their coverage, especially in terms of high levels of protection, adopting a precautionary pause on deep‐sea mining, ending overfishing and destructive fishing practices, radically reducing marine pollution, putting in place a financing mechanism for ocean management and protection, and lastly, scaling up science/data gathering and facilitating data sharing.
5. By implementing all eight measures in unison, as a coordinated strategy, we can build resilience to climate change, help sustain fisheries productivity, particularly for low‐income countries dependent on fisheries, protect coasts (e.g. via soft‐engineering/habitat‐based approaches), promote mitigation (e.g. carbon storage) and enable improved adaptation to rapid global change.

     

Antigen-specific proliferation and activation of peripheral blood mononuclear cells from Mycobacterium bovis-infected reindeer

To evaluate antigen-specific proliferative and activation-associated responses from Mycobacterium bovis-infected reindeer, blood mononuclear cells from M. bovis- (n = 10) and non-infected reindeer (n = 4) were stimulated with a recombinant early secretory antigenic target-6 and culture filtrate protein-10 fusion protein (rESAT6:CFP10), M. bovis purified protein derivative, pokeweed mitogen, or medium alone and evaluated by flow cytometry using dye tracker analysis and cell surface marker staining. gammadelta TCR+ and CD8+ cells, but not CD4+ cells, from M. bovis-infected reindeer proliferated in response to specific antigen stimulation. Expression (i.e., mean fluorescence intensity) of CD44 was increased and CD62L decreased on proliferative as compared to non-proliferative fractions in antigen- and mitogen-stimulated cultures. In response rESAT6:CFP10 stimulation, MHC II fluorescence intensity was increased on CD4+, gammadelta TCR+, CD172a+, and IgM+ cells from infected reindeer as compared to that of non-stimulated cells from the same reindeer. Recombinant ESAT6:CFP10 stimulation also induced expansion of a CD172a+, MHC II+ population within mononuclear cell cultures from M. bovis-infected reindeer. Despite a moderate challenge dose and extended duration of incubation, experimental infection of reindeer was generally limited to lymph nodes draining the inoculation site, suggestive of host resistance to progressive disease. Present in vitro findings, therefore, may be predictive of host responses by reindeer that limit progression to disseminated disease.     

The benthic impacts of a large cod farm — Are there indicators for environmental sustainability?

A cod farm in Vidlin Voe, Shetland was at the time of the study the largest cod farm in Europe, and was started in 2003. Run according to organic principles, the farm consisted of 17  32 m diameter cages in the 3 km long sea loch (locally known as voe), which though extremely exposed at the mouth, is sheltered near the cages.Benthic macrofauna and physical/chemical sediment parameters were measured in the summer months of 2004, 2005 and 2006 as biomass increased in the growing cycle. A range of biotic indices showed the benthic environment was enriched at peak biomass of the farm, but by contrast, surficial CHN levels and organic matter were low and uniform. The commonly used sediment indicators redox, total organic carbon and total organic nitrogen did not correlate well with biotic indicators at this site. Predictions from a near-field particle tracking and resuspension model (CODMOD) and a simple nutrient enhancement box model of the voe system (Equilibrium Concentration Enhancement (ECE) model), were used to assess the assimilative capacity of the system.