Real time detectionof airborne fungal spores and investigations into their dynamics in indoor air

Concern regarding the health effects of indoor air quality has grown in recent years, due to the increased prevalence of many diseases, as well as the fact that many people now spend most of their time indoors. While numerous studies have reported on the dynamics of aerosols indoors, the dynamics of bioaerosols in indoor environments are still poorly understood and very few studies have focused on fungal spore dynamics in indoor environments. Consequently, this work investigated the dynamics of fungal spores in indoor air, including fungal spore release and deposition, as well as investigating the mechanisms involved in the fungal spore fragmentation process. In relation to the investigation of fungal spore dynamics, it was found that the deposition rates of the bioaerosols (fungal propagules) were in the same range as the deposition rates of nonbiological particles and that they were a function of their aerodynamic diameters. It was also found that fungal particle deposition rates increased with increasing ventilation rates. These results (which are reported for the first time) are important for developing an understanding of the dynamics of fungal spores in the air. In relation to the process of fungal spore fragmentation, important information was generated concerning the airborne dynamics of the spores, as well as the part/s of the fungi which undergo fragmentation. The results obtained from these investigations into the dynamics of fungal propagules in indoor air significantly advance knowledge about the fate of fungal propagules in indoor air, as well as their deposition in the respiratory tract. The need to develop an advanced, real-time method for monitoring bioaerosols has become increasingly important in recent years, particularly as a result of the increased threat from biological weapons and bioterrorism. However, to date, the Ultraviolet Aerodynamic Particle Sizer (UVAPS, Model 3312, TSI, St Paul, MN) is the only commercially available instrument capable of monitoring and measuring viable airborne micro-organisms in real-time. Therefore (for the first time), this work also investigated the ability of the UVAPS to measure and characterise fungal spores in indoor air. The UVAPS was found to be sufficiently sensitive for detecting and measuring fungal propagules. Based on fungal spore size distributions, together with fluorescent percentages and intensities, it was also found to be capable of discriminating between two fungal spore species, under controlled laboratory conditions. In the field, however, it would not be possible to use the UVAPS to differentiate between different fungal spore species because the different micro-organisms present in the air may not only vary in age, but may have also been subjected to different environmental conditions. In addition, while the real-time UVAPS was found to be a good tool for the investigation of fungal particles under controlled conditions, it was not found to be selective for bioaerosols only (as per design specifications). In conclusion, the UVAPS is not recommended for use in the direct measurement of airborne viable bioaerosols in the field, including fungal particles, and further investigations into the nature of the micro-organisms, the UVAPS itself and/or its use in conjunction with other conventional biosamplers, are necessary in order to obtain more realistic results. Overall, the results obtained from this work on airborne fungal particle dynamics will contribute towards improving the detection capabilities of the UVAPS, so that it is capable of selectively monitoring and measuring bioaerosols, for which it was originally designed. This work will assist in finding and/or improving other technologies capable of the real-time monitoring of bioaerosols. The knowledge obtained from this work will also be of benefit in various other bioaerosol applications, such as understanding the transport of bioaerosols indoors.

Application of conventional and real-time fluorescent ITS1 rDNA PCR for detection of Besnoitia besnoiti infections in bovine skin biopsies

Besnoitia besnoiti, an apicomplexan protozoan parasite, is the causative agent of bovine besnoitiosis. This infection may dramatically affect body condition, and, in males, lead to irreversible infertility. While identification of clinical cases and their histopathological confirmation is relatively simple to carry out, finding subclinical forms of infection is more difficult, thus a more sensitive test for the identification of the etiological agent may be an appropriate diagnostic tool. We have developed the ITS1 rDNA-sequence-based conventional and real-time PCR which are highly sensitive and specific for the detection of B. besnoiti infection in cattle. A recombinant internal positive control was introduced to assess possible sample-related inhibitory effects during the amplification reaction and, in order to prevent false-positive results, a pre-PCR treatment of potentially contaminating dU-containing PCR product with uracil-DNA-glycosylase (UDG) was followed.

Application of real-time fluorescent PCR for quantitative assessment of Neospora caninum infections in organotypic slice cultures of rat central nervous system tissue

The previously described Nc5-specific PCR test for the diagnosis of Neospora caninum infections was used to develop a quantitative PCR assay which allows the determination of infection intensities within different experimental and diagnostic sample groups. The quantitative PCR was performed by using a dual fluorescent hybridization probe system and the LightCycler Instrument for online detection of amplified DNA. This assay was successfully applied for demonstrating the parasite proliferation kinetics in organotypic slice cultures of rat brain which were infected in vitro with N. caninum tachyzoites. This PCR-based method of parasite quantitation with organotypic brain tissue samples can be regarded as a novel ex vivo approach for exploring different aspects of cerebral N. caninum infection.

Difterentially expressed genes of Tetrahymena thermophila in response to tributyltin (TBT) identified by suppression subtractive hybridization and real time quantitative PCR

Tributyltin (TBT) is widely used as antifouling paints, agriculture biocides, and plastic stabilizers around the world, resulting in great pollution problem in aquatic environments. However, it has been short of the biomonitor to detect TBT in freshwater. We constructed the suppression subtractive hybridization library of Tetrahymena thermophila exposed to TBT, and screened out 101 Expressed Sequence Tags whose expressions were significantly up- or down-regulated with TBT treatment. From this, a series of genes related to the TBT toxicity were discovered, such as glutathione-S-transferase gene (down-regulated), plasma membrane Ca2+ ATPase isoforms 3 gene (up-regulated) and NgoA (up-regulated). Furthermore, their expressions under different concentrations of TBT treatment (0.5-40 ppb) were detected by real time fluorescent quantitative PCR. The differentially expressed genes of T thermophila in response to TBT were identified, which provide the basic to make Tetrahymena as a sensitive, rapid and convenient TBT biomonitor in freshwater based on rDNA inducible expression system. (c) 2006 Elsevier B.V. All rights reserved.

Size distributions and temporal variations of biological aerosol particles in the Amazon rainforest characterized by microscopy and real-time UV-APS fluorescence techniques during AMAZE-08

As a part of the AMAZE-08 campaign during the wet season in the rainforest of central Amazonia, an ultraviolet aerodynamic particle sizer (UV-APS) was operated for continuous measurements of fluorescent biological aerosol particles (FBAP). In the coarse particle size range (> 1 mu m) the campaign median and quartiles of FBAP number and mass concentration were 7.3x10(4) m(-3) (4.0-13.2x10(4) m(-3)) and 0.72 mu g m(-3) (0.42-1.19 mu g m(-3)), respectively, accounting for 24% (11-41%) of total particle number and 47% (25-65%) of total particle mass. During the five-week campaign in February-March 2008 the concentration of coarse-mode Saharan dust particles was highly variable. In contrast, FBAP concentrations remained fairly constant over the course of weeks and had a consistent daily pattern, peaking several hours before sunrise, suggesting observed FBAP was dominated by nocturnal spore emission. This conclusion was supported by the consistent FBAP number size distribution peaking at 2.3 mu m, also attributed to fungal spores and mixed biological particles by scanning electron microscopy (SEM), light microscopy and biochemical staining. A second primary biological aerosol particle (PBAP) mode between 0.5 and 1.0 mu m was also observed by SEM, but exhibited little fluorescence and no true fungal staining. This mode may have consisted of single bacterial cells, brochosomes, various fragments of biological material, and small Chromalveolata (Chromista) spores. Particles liquid-coated with mixed organic-inorganic material constituted a large fraction of observations, and these coatings contained salts likely from primary biological origin. We provide key support for the suggestion that real-time laser-induce fluorescence (LIF) techniques using 355 nm excitation provide size-resolved concentrations of FBAP as a lower limit for the atmospheric abundance of biological particles in a pristine environment. We also show some limitations of using the instrument for ambient monitoring of weakly fluorescent particles < 2 mu m. Our measurements confirm that primary biological particles, fungal spores in particular, are an important fraction of supermicron aerosol in the Amazon and that may contribute significantly to hydrological cycling, especially when coated by mixed inorganic material.

On Real-Time Monitoring And Control For Efficient Management Of Drinking Water Networks: Barcelona Case Study

Drinking water utilities in urban areas are focused on finding smart solutions facing new challenges in their real-time operation because of limited water resources, intensive energy requirements, a growing population, a costly and ageing infrastructure, increasingly stringent regulations, and increased attention towards the environmental impact of water use. Such challenges force water managers to monitor and control not only water supply and distribution, but also consumer demand. This paper presents and discusses novel methodologies and procedures towards an integrated water resource management system based on advanced ICT technologies of automation and telecommunications for largely improving the efficiency of drinking water networks (DWN) in terms of water use, energy consumption, water loss minimization, and water quality guarantees. In particular, the paper addresses the first results of the European project EFFINET (FP7-ICT2011-8-318556) devoted to the monitoring and control of the DWN in Barcelona (Spain). Results are split in two levels according to different management objectives: (i) the monitoring level is concerned with all the aspects involved in the observation of the current state of a system and the detection/diagnosis of abnormal situations. It is achieved through sensors and communications technology, together with mathematical models; (ii) the control level is concerned with computing the best suitable and admissible control strategies for network actuators as to optimize a given set of operational goals related to the performance of the overall system. This level covers the network control (optimal management of water and energy) and the demand management (smart metering, efficient supply). The consideration of the Barcelona DWN as the case study will allow to prove the general applicability of the proposed integrated ICT solutions and their effectiveness in the management of DWN, with considerable savings of electricity costs and reduced water loss while ensuring the high European standards of water quality to citizens.

Development of a real time Image analysis system for traffic monitoring applications.

Image processing offers unparalleled potential for traffic monitoring and control. For many years engineers have attempted to perfect the art of automatic data abstraction from sequences of video images. This paper outlines a research project undertaken at Napier University by the authors in the field of image processing for automatic traffic analysis. A software based system implementing TRIP algorithms to count cars and measure vehicle speed has been developed by members of the Transport Engineering Research Unit (TERU) at the University. The TRIP algorithm has been ported and evaluated on an IBM PC platform with a view to hardware implementation of the pre-processing routines required for vehicle detection. Results show that a software based traffic counting system is realisable for single window processing. Due to the high volume of data required to be processed for full frames or multiple lanes, system operations in real time are limited. Therefore specific hardware is required to be designed. The paper outlines a hardware design for implementation of inter-frame and background differencing, background updating and shadow removal techniques. Preliminary results showing the processing time and counting accuracy for the routines implemented in software are presented and a real time hardware pre-processing architecture is described.

Scene invariant crowd counting for real-time surveillance

Automated crowd counting allows excessive crowding to be detected immediately, without the need for constant human surveillance. Current crowd counting systems are location specific, and for these systems to function properly they must be trained on a large amount of data specific to the target location. As such, configuring multiple systems to use is a tedious and time consuming exercise. We propose a scene invariant crowd counting system which can easily be deployed at a different location to where it was trained. This is achieved using a global scaling factor to relate crowd sizes from one scene to another. We demonstrate that a crowd counting system trained at one viewpoint can achieve a correct classification rate of 90% at a different viewpoint.

A criminological analysis : using real-time monitoring to gather data on online predators

The Internet presents a constantly evolving frontier for criminology and policing, especially in relation to online predators – paedophiles operating within the Internet for safer access to children, child pornography and networking opportunities with other online predators. The goals of this qualitative study are to undertake behavioural research – identify personality types and archetypes of online predators and compare and contrast them with behavioural profiles and other psychological research on offline paedophiles and sex offenders. It is also an endeavour to gather intelligence on the technological utilisation of online predators and conduct observational research on the social structures of online predator communities. These goals were achieved through the covert monitoring and logging of public activity within four Internet Relay Chat(rooms) (IRC) themed around child sexual abuse and which were located on the Undernet network. Five days of monitoring was conducted on these four chatrooms between Wednesday 1 to Sunday 5 April 2009; this raw data was collated and analysed. The analysis identified four personality types – the gentleman predator, the sadist, the businessman and the pretender – and eight archetypes consisting of the groomers, dealers, negotiators, roleplayers, networkers, chat requestors, posters and travellers. The characteristics and traits of these personality types and archetypes, which were extracted from the literature dealing with offline paedophiles and sex offenders, are detailed and contrasted against the online sexual predators identified within the chatrooms, revealing many similarities and interesting differences particularly with the businessman and pretender personality types. These personality types and archetypes were illustrated by selecting users who displayed the appropriate characteristics and tracking them through the four chatrooms, revealing intelligence data on the use of proxies servers – especially via the Tor software – and other security strategies such as Undernet’s host masking service. Name and age changes, which is used as a potential sexual grooming tactic was also revealed through the use of Analyst’s Notebook software and information on ISP information revealed the likelihood that many online predators were not using any safety mechanism and relying on the anonymity of the Internet. The activities of these online predators were analysed, especially in regards to child sexual grooming and the ‘posting’ of child pornography, which revealed a few of the methods in which online predators utilised new Internet technologies to sexually groom and abuse children – using technologies such as instant messengers, webcams and microphones – as well as store and disseminate illegal materials on image sharing websites and peer-to-peer software such as Gigatribe. Analysis of the social structures of the chatrooms was also carried out and the community functions and characteristics of each chatroom explored. The findings of this research have indicated several opportunities for further research. As a result of this research, recommendations are given on policy, prevention and response strategies with regards to online predators.

Real-time risk monitoring in business processes : a sensor-based approach

This article proposes an approach for real-time monitoring of risks in executable business process models. The approach considers risks in all phases of the business process management lifecycle, from process design, where risks are defined on top of process models, through to process diagnosis, where risks are detected during process execution. The approach has been realized via a distributed, sensor-based architecture. At design-time, sensors are defined to specify risk conditions which when fulfilled, are a likely indicator of negative process states (faults) to eventuate. Both historical and current process execution data can be used to compose such conditions. At run-time, each sensor independently notifies a sensor manager when a risk is detected. In turn, the sensor manager interacts with the monitoring component of a business process management system to prompt the results to process administrators who may take remedial actions. The proposed architecture has been implemented on top of the YAWL system, and evaluated through performance measurements and usability tests with students. The results show that risk conditions can be computed efficiently and that the approach is perceived as useful by the participants in the tests.