Evaluation of the MIDTAL microarray chip for monitoring toxic microalgae in the Orkney Islands, U.K.

Harmful or nuisance algal blooms can cause economic damage to fisheries and tourism. Additionally, toxins produced by harmful algae and ingested via contaminated shellfish can be potentially fatal to humans. The seas around the Orkney Islands, UK currently hold a number of toxic algal species which cause shellfishery closures in most years. Extensive and costly monitoring programs are carried out to detect harmful microalgae before they reach action levels. However, the ability to distinguish between toxic and non-toxic strains of some algae is not possible using these methods. The microarrays for the detection of toxic algae (MIDTAL) microarray contains rRNA probes for toxic algal species/strains which have been adapted and optimized for microarray use. In order to investigate the use of the chip for monitoring in the Orkney Islands, samples were collected between 2009 and 2011 from Brings Deep, Scapa Flow, Orkney Islands, UK; RNA was extracted and hybridized with generation 2 and 3.1 of the chip. The data were then compared to cell counts performed under light microscopy and in the case of Alexandrium tamarense to qPCR data targeting the saxitoxin gene and the LSU-rRNA gene. A good agreement between cell numbers and microarray signal was found for A. tamarense, Pseudo-nitzschia sp., Dinophysis sp. (r?<?0.5, for all) in addition to this there the chip successfully detected a large bloom of Karenia mikimotoi (r?<?0.70) in August and September 2011. Overall, there was good improvement in probe signal between generation 2 and generation 3.1 of the chip with much less variability and more consistent results and better correlation between the probes. The chip performed well for A. tamarense group I signal to cell numbers in calibrations (r?>?0.9). However, in field samples, this correlation was slightly lower suggesting interactions between all species in the sample may affect signal. Overall, the chip showed it could identify the presence of target species in field samples although some work is needed to improve the quantitative nature of the chip before it would be suitable for monitoring in the Orkney Islands.     

Molecular probes and microarrays for the detection of toxic algae in the genera Dinophysis and Phalacroma (Dinophyta)

Dinophysis and Phalacroma species containing diarrheic shellfish toxins and pectenotoxins occur in coastal temperate waters all year round and prevent the harvesting of mussels during several months each year in regions in Europe, Chile, Japan, and New Zealand. Toxicity varies among morphologically similar species, and a precise identification is needed for early warning systems. Molecular techniques using ribosomal DNA sequences offer a means to identify and detect precisely the potentially toxic species. We designed molecular probes targeting the 18S rDNA at the family and genus levels for Dinophysis and Phalacroma and at the species level for Dinophysis acuminata, Dinophysis acuta, and Dinophysis norvegica, the most commonly occurring, potentially toxic species of these genera in Western European waters. Dot blot hybridizations with polymerase chain reaction (PCR)-amplified rDNA from 17 microalgae were used to demonstrate probe specificity. The probes were modified along with other published fluorescence in situ hybridization and PCR probes and tested for a microarray platform within the MIDTAL project (http://www.midtal.com). The microarray was applied to field samples from Norway and Spain and compared to microscopic cell counts. These probes may be useful for early warning systems and monitoring and can also be used in population dynamic studies to distinguish species and life cycle stages, such as cysts, and their distribution in time and space.     

Determination of organochlorine pesticide residue in sediment and water from the Densu river basin, Ghana

The distribution of organochlorine pesticides in the aquatic ecosystem from the Densu river revealed varying levels of concentration in water and the sediment samples. Three locations were sampled along the river to evaluate the levels of organochlorine pesticide residue in the river. Sediment and surface water samples were extracted by soxhlet and liquid-liquid extraction respectively and analyzed using Gas Chromatograph coupled with electron capture detector. The detectable organochlorine pesticides were gamma-hexachlorocyclohexane (HCH), delta-hexachlorocyclohexane, heptachlor, aldrin and dieldrin. The other pesticides that were investigated are gamma-chlordane, alpha endosulfan, endosulfan sulfate, p,p′-DDT and its metabolite p,p′-DDE, methoxychlor, endrin and its metabolite endrin aldehyde and endrin ketone. The order of increasing frequency of detection of samples was higher in sediment than water. In sediment, the mean concentration ranged from 0.030 μg kg⁻¹ dry weight (endrin) to 10.98 μg kg⁻¹ dry weight (aldrin). The highest detected concentration of organochlorine in water was endosulfan sulfate with mean concentration of 0.185 μg L⁻¹. Analysis of variance indicated significant differences for most organochlorine pesticide residue in the sediment sampled from the various locations. Some of the levels of organochlorine pesticides detected in water were relatively high compared to guideline values set by World Health Organization and Australia and thus could be harmful if the trend is not checked.     

Importance of conserving large and old trees to continuity of tree-related microhabitats

Protecting structural features, such as tree-related microhabitats (TreMs), is a cost-effective tool crucial for biodiversity conservation applicable to large forested landscapes. Although the development of TreMs is influenced by tree diameter, species, and vitality, the relationships between tree age and TreM profile remain poorly understood. Using a tree-ring-based approach and a large data set of 8038 trees, we modeled the effects of tree age, diameter, and site characteristics on TreM richness and occurrence across some of the most intact primary temperate forests in Europe, including mixed beech and spruce forests. We observed an overall increase in TreM richness on old and large trees in both forest types. The occurrence of specific TreM groups was variably related to tree age and diameter, but some TreM groups (e.g., epiphytes) had a stronger positive relationship with tree species and elevation. Although many TreM groups were positively associated with tree age and diameter, only two TreM groups in spruce stands reacted exclusively to tree age (insect galleries and exposed sapwood) without responding to diameter. Thus, the retention of trees for conservation purposes based on tree diameter appears to be a generally feasible approach with a rather low risk of underrepresentation of TreMs. Because greater tree age and diameter positively affected TreM development, placing a greater emphasis on conserving large trees and allowing them to reach older ages, for example, through the establishment of conservation reserves, would better maintain the continuity of TreM resource and associated biodiversity. However, this approach may be difficult due to the widespread intensification of forest management and global climate change.     

Seasonal dynamics of freshwater pathogens as measured by microarray at Lake Sapanca,a drinking water source in the north-eastern part of Turkey

Monitoring drinking water quality is an important public health issue. Two objectives from the 4 years, six nations, EU Project μAqua were to develop hierarchically specific probes to detect and quantify pathogens in drinking water using a PCR-free microarray platform and to design a standardised water sampling program from different sources in Europe to obtain sufficient material for downstream analysis. Our phylochip contains barcodes (probes) that specifically identify freshwater pathogens that are human health risks in a taxonomic hierarchical fashion such that if species is present, the entire taxonomic hierarchy (genus, family, order, phylum, kingdom) leading to it must also be present, which avoids false positives. Molecular tools are more rapid, accurate and reliable than traditional methods, which means faster mitigation strategies with less harm to humans and the community. We present microarray results for the presence of freshwater pathogens from a Turkish lake used drinking water and inferred cyanobacterial cell equivalents from samples concentrated from 40 into 1 L in 45 min using hollow fibre filters. In two companion studies from the same samples, cyanobacterial toxins were analysed using chemical methods and those dates with highest toxin values also had highest cell equivalents as inferred from this microarray study.     

Evaluation of a high visibility enforcement project focused on passenger vehicles interacting with commercial vehicles

Introduction

In 2004, Washington State applied NHTSA's High Visibility Enforcement model used in the Click It or Ticket seat belt campaign in an attempt to reduce unsafe driving behaviors around commercial motor vehicles (CMVs). The program was called Ticketing Aggressive Cars and Trucks (TACT). This paper details the methods used to evaluate the program's effectiveness and the results of the evaluation.

Method

Four high-crash interstate highway corridors, each approximately 25 miles in length, were selected. Two of these corridors received TACT media messages and increased enforcement over an 18-month period while two comparison corridors did not receive any increased media or enforcement.

Results

A total of 4,737 contacts were made with drivers during the two enforcement waves, and 72% of these contacts led to a citation. Drivers at the intervention sites who said they saw or heard any of the TACT messages increased from 17.7% in the pre period to a high of 67.3% in the post periods. Drivers at the intervention sites also reported increased exposure to the core message of leaving more space when passing trucks (14% pre to 40% post period). The percentage of drivers who said they leave more room when passing trucks than when passing cars rose from 16% in the pre period to 24% in the post period at the intervention sites, while comparison sites showed no change. Over 150 hours of video recorded by law enforcement officers in unmarked vehicles were utilized to examine violation rates and severity of violations before and after the intervention campaigns. Statistical analyses showed that violation rates were reduced significantly at the intervention sites (between 23% and 46%), while remaining constant at the comparison sites. Analyses of the video data also showed that the seriousness of the residual violations at the intervention sites decreased.

Conclusions

Overall, the evaluation results provide a consistent picture of the effectiveness of the TACT pilot project. Success was demonstrated at every step - messages were received and understood, knowledge was changed in the intended direction, self reported driving behavior around large trucks improved, and observed driving behaviors confirmed the self reports.

Impact on Industry

After this initial success in Washington State, the TACT model will continue to be implemented and evaluated by FMCSA in an attempt to validate the program. Based on the results of this study and the consistent positive results found for other sTEP projects, it is likely that TACT will show continued success in a variety of settings and will help reduce the number and severity of crashes involving CMVs. Future research should attempt to use many of the methods described here to further validate the methods for not only evaluations of TACT programs, but also for any other highway safety programs that require measurements of the program's effectiveness.     

Facilitating the improved management of waste in South Africa through a national waste information system

Developing a waste information system (WIS) for a country is more than just about collecting routine data on waste; it is about facilitating the improved management of waste by providing timely, reliable information to the relevant role-players. It is a means of supporting the waste governance challenges facing South Africa - challenges ranging from strategic waste management issues at national government to basic operational challenges at local government. The paper addresses two hypotheses. The first is that the identified needs of government can provide a platform from which to design a national WIS framework for a developing country such as South Africa, and the second is that the needs for waste information reflect greater, currently unfulfilled challenges in the sustainable management of waste. Through a participatory needs analysis process, it is shown that waste information is needed by the three spheres of government, to support amongst others, informed planning and decision-making, compliance monitoring and enforcement, community participation through public access to information, human, infrastructure and financial resource management and policy development. These needs for waste information correspond closely with key waste management challenges currently facing the country. A shift in governments approach to waste, in line with national and international policy, is evident from identified current and future waste information needs. However, the need for information on landfilling remains entrenched within government, possibly due to the poor compliance of landfill sites in South Africa and the problems around the illegal disposal of both general and hazardous waste.     

Indicators for managing biosolids in Ireland

Sustainable development indicators (SDIs) have emerged as a tool to measure progress towards sustainable development for a number of fields. However, no indicator initiative to date has been aimed at biosolids management at local authority, regional or national levels. This paper presents a study where stakeholders involved in the management of biosolids in Ireland participated in the development of SDIs for managing biosolids at the local/regional level. A significant 81% of participating stakeholders find SDIs either 'useful' or 'very useful' as a tool for managing biosolids. A suite of 22 indicators has been developed and arranged according to the driving force-pressure-state-impact-response (DPSIR) indicator framework. The indicators address all the domains of biosolids management namely, production, quality, cost, legislation/regulation, training/research and recycling/disposal. The stakeholder approach is recognition that no effective indicator set can be developed without the input of stakeholders.     

The efficiency of using non-culpable crash-claim involvements from insurance data as a means of estimating travel exposure for road user sub-groups

Introduction

Induced exposure has a long history of development and usage in traffic safety research but a major question has always concerned the extent to which the accumulation of culpable and non-culpable involvements can be considered independent.

Method

Culpability assessments of 32,630 vehicles' crash-claim involvements adjudicated by insurance adjusters were matched with vehicle odometer readings taken at emission testing using consistent identification of vehicles and principal operators over a 5-year period.

Result

It was found that the accumulation of culpable crash involvements was not entirely independent of that for non-culpable involvements. However, the rate of non-culpable involvements was determined to be an acceptable surrogate for travel exposure rate where sample sizes were large.

Discussion

The relationship between the rate of non-culpable involvements and the rate of travel exposure for data subsets when both were normalized by the overall sample rates was reminiscent of an accident-volume curve for roadway locations in traffic engineering theory. This suggested that only a portion of non-culpable involvements actually related directly to travel and this lead to a correction factor that could be applied.

Impact on Industry

While lack of independence of involvement rates may be problematic for a direct risk ratio application, it does not invalidate the use of non-culpable involvements to predict travel. For insurers that have a need to estimate travel amounts for different driver/vehicle groups as part of the insurance rating purposes, this can be a useful application.