Food Safety Testing: Rapid Molecular Methods for Chemical and Biological Hazards

The central goal of food safety policy in the European Union (EU) is to protect consumer health by guaranteeing a high level of food safety throughout the food chain. This goal can in part be achieved by testing foodstuffs for the presence of various chemical and biological hazards. The aim of this study was to facilitate food safety testing by providing rapid and user-friendly methods for the detection of particular food-related hazards. Heterogeneous competitive time-resolved fluoroimmunoassays were developed for the detection of selected veterinary residues, that is coccidiostat residues, in eggs and chicken liver. After a simplified sample preparation procedure, the immunoassays were performed either in manual format with dissociation-enhanced measurement or in automated format with pre-dried assay reagents and surface measurement. Although the assays were primarily designed for screening purposes providing only qualitative results, they could also be used in a quantitative mode. All the developed assays had good performance characteristics enabling reliable screening of samples at concentration levels required by the authorities. A novel polymerase chain reaction (PCR)-based assay system was developed for the detection of Salmonella spp. in food. The sample preparation included a short non-selective pre-enrichment step, after which the target cells were collected with immunomagnetic beads and applied to PCR reaction vessels containing all the reagents required for the assay in dry form. The homogeneous PCR assay was performed with a novel instrument platform, GenomEra^™, and the qualitative assay results were automatically interpreted based on end-point time-resolved fluorescence measurements and cut-off values. The assay was validated using various food matrices spiked with sub-lethally injured Salmonella cells at levels of 1-10 colony forming units (CFU)/25 g of food. The main advantage of the system was the exceptionally short time to result; the entire process starting from the pre-enrichment and ending with the PCR result could be completed in eight hours. In conclusion, molecular methods using state-of-the-art assay techniques were developed for food safety testing. The combination of time-resolved fluorescence detection and ready-to-use reagents enabled sensitive assays easily amenable to automation. Consequently, together with the simplified sample preparation, these methods could prove to be applicable in routine testing.

Tuotannonohjauksen kehittäminen piensarjatuotannossa

Tämä diplomityö käsittelee teollisen yrityksen tuotannonohjauksen kehittämistä piensarjatuotannossa. Työn kohteena on ABB Oy:n Tuulivoimageneraattorit-tulosyksikkö, joka valmistaa vakiotuotteita asiakasohjautuvasti. Työssä esitellään aluksi tuotannon ja tuotannonohjauksen teoriaa. Lävitse käydään perusasioiden kuten määritelmien, tavoitteiden ja tehtävien lisäksi tuotannonohjausprosessia sekä tuotannonohjauksen tietotekniikkaa. Teorian jälkeisessä empiriaosuudessa esitellään työssä kehitettyjä keinoja tuotannonohjauksen parantamiseksi. Tutkimus on toteutettu teoreettisen ja empiirisen tutkimustyön avulla. Teoreettiseen tutkimustyöhön sisältyi suomalaisiin ja ulkomaalaisiin kirjallisuuslähteisiin perehtyminen. Empiirinen tutkimustyö suoritettiin itsenäisen ongelman ratkaisutyön avulla. Tämä sisälsi kehittämiskohteiden analysoinnin, tarkempien kehittämistarpeiden määrityksen sekä kokeilujen kautta tapahtuneen kehittämistyön. Tutkimuksen päätavoitteena oli selvittää, miten tuotannonohjauksen kehittämisellä voidaan parantaa kohteena olevan tulosyksikön tuottavuutta ja kannattavuutta. Päätavoitteen pohjalta muodostettiin kuusi osatavoitetta: toimitusvarmuuden parantaminen, kapasiteetin kuormitusasteen nostaminen, kapasiteetin suunnittelun kehittäminen, läpäisyaikojen lyhentäminen, uuden ERP-järjestelmän vaatimusmäärittely sekä tuotannonohjausprosessin määrittäminen. Työssä rakennettiin neljään ensiksi mainittuun osatavoitteeseen tietotekniset sovellukset, jotka mahdollistavat osatavoitteiden suunnittelun ja ohjaamisen. Sovelluksia varten kullekin tuotteelle määriteltiin esimerkiksi työnvaiheketjut läpäisyaikoineen, kuormitusryhmät, kuormitusryhmien kapasiteetit, tuotteiden kuormittavuudet sekä kriittiset työvälineet. Työ osoitti, että tietotekniikka auttaa suuresti tuotannonohjauksessa. Lisääntynyt läpinäkyvyys, parantunut tiedonkulku, simulointimahdollisuudet sekä graafinen esitystapa helpottavat erilaisten suunnitelmien teossa ja parantavat siten päätöksenteon laatua. Tietotekniikan hyväksikäytön pohjana toimii tuotannon perus- ja tapahtumatietojen kurinalainen päivitys. Tämän vuoksi tietojärjestelmistä kannattaa rakentaa mahdollisimman yksinkertaisia.

ZigBee-verkot ja evaluointilaitteen suunnittelu

In this thesis a closer look at ZigBee wireless network protocol was made and a ZigBee evaluation device was designed and built for testing it. ZigBee is a digital wireless network technology published in 2004, which was developed for wireless control and automation systems. It is based on the IEEE 802.15.4-2003 standard, and its aim was to create a reliable wireless networking protocol with secure data transmissions, low power consumption and low manufacturing costs. The low power consumption requirement has also led to a slow data rate and a short range, but in a ZigBee network individual network devices can route messages to each other, so short range is not a problem. In automation and control systems a fast data rate is not usually an important requirement either. ZigBee network can form automatically and one ZigBee network can theoretically include thousands of devices, all of which can communicate with each other. ZigBee network is also self-healing, so an individual device dropping out of the network, like breaking down, does not affect functionality of the network. Evaluation device was designed and implemented in four different versions, which were tested with six different antenna designs. All antennas were tested at a range of 100 meters in an open terrain using two different frequencies and transmission powers. Also a range test was performed for selected antennas using transmission power of 4,5 dBm, which was maximum power of the ZigBee chip used in the evaluation device. Range test resulted in a measured range of several hundred meters, which can be considered as a good result for a short-range technology.

Frequency-Domain and Wide-Pulse Time-Domain Measurements of Lanthanide Luminescence and Lanthanide-Based Resonance Energy Transfer

Resonance energy transfer (RET) is a non-radiative transfer of the excitation energy from the initially excited luminescent donor to an acceptor. The requirements for the resonance energy transfer are: i) the spectral overlap between the donor emission spectrum and the acceptor absorption spectrum, ii) the close proximity of the donor and the acceptor, and iii) the suitable relative orientations of the donor emission and the acceptor absorption transition dipoles. As a result of the RET process the donor luminescence intensity and the donor lifetime are decreased. If the acceptor is luminescent, a sensitized acceptor emission appears. The rate of RET depends strongly on the donor–acceptor distance (r) and is inversely proportional to r6. The distance dependence of RET is utilized in binding assays. The proximity requirement and the selective detection of the RET-modified emission signal allow homogeneous separation free assays. The term lanthanide-based RET is used when luminescent lanthanide compounds are used as donors. The long luminescence lifetimes, the large Stokes’ shifts and the intense, sharply-spiked emission spectra of the lanthanide donors offer advantages over the conventional organic donor molecules. Both the organic lanthanide chelates and the inorganic up-converting phosphor (UCP) particles have been used as donor labels in the RET based binding assays. In the present work lanthanide luminescence and lanthanide-based resonance energy transfer phenomena were studied. Luminescence lifetime measurements had an essential role in the research. Modular frequency-domain and time-domain luminometers were assembled and used successfully in the lifetime measurements. The frequency-domain luminometer operated in the low frequency domain ( 100 kHz) and utilized a novel dual-phase lock-in detection of the luminescence. One of the studied phenomena was the recently discovered non-overlapping fluorescence resonance energy transfer (nFRET). The studied properties were the distance and temperature dependences of nFRET. The distance dependence was found to deviate from the Förster theory and a clear temperature dependence was observed whereas conventional RET was completely independent of the temperature. Based on the experimental results two thermally activated mechanisms were proposed for the nFRET process. The work with the UCP particles involved the measurement of the luminescence properties of the UCP particles synthesized in our laboratory. The goal of the UCP particle research is to develop UCP donor labels for binding assays. In the present work the effect of the dopant concentrations and the core–shell structure on the total up-conversion luminescence intensity, the red–green emission ratio, and the luminescence lifetime was studied. Also the non-radiative nature of the energy transfer from the UCP particle donors to organic acceptors was demonstrated for the first time in aqueous environment and with a controlled donor–acceptor distance.

Characterization and treatment of sisal fiber residues for cement-based composite application

Sisal fiber is an important agricultural product used in the manufacture of ropes, rugs and also as a reinforcement of polymeric or cement-based composites. However, during the fiber production process a large amount of residues is generated which currently have a low potential for commercial use. The aim of this study is to characterize the agricultural residues by the production and improvement of sisal fiber, called field bush and refugo and verify the potentiality of their use in the reinforcement of cement-based composites. The residues were treated with wet-dry cycles and evaluated using tensile testing of fibers, scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectroscopy. Compatibility with the cement-based matrix was evaluated through the fiber pull-out test and flexural test in composites reinforced with 2 % of sisal residues. The results indicate that the use of treated residue allows the production of composites with good mechanical properties that are superior to the traditional composites reinforced with natural sisal fibers.

Monte Carlo Hypothesis Testing with The Sharpe Ratio

The purpose of this master thesis was to perform simulations that involve use of random number while testing hypotheses especially on two samples populations being compared weather by their means, variances or Sharpe ratios. Specifically, we simulated some well known distributions by Matlab and check out the accuracy of an hypothesis testing. Furthermore, we went deeper and check what could happen once the bootstrapping method as described by Effrons is applied on the simulated data. In addition to that, one well known RobustSharpe hypothesis testing stated in the paper of Ledoit and Wolf was applied to measure the statistical significance performance between two investment founds basing on testing weather there is a statistically significant difference between their Sharpe Ratios or not. We collected many literatures about our topic and perform by Matlab many simulated random numbers as possible to put out our purpose; As results we come out with a good understanding that testing are not always accurate; for instance while testing weather two normal distributed random vectors come from the same normal distribution. The Jacque-Berra test for normality showed that for the normal random vector r1 and r2, only 94,7% and 95,7% respectively are coming from normal distribution in contrast 5,3% and 4,3% failed to shown the truth already known; but when we introduce the bootstrapping methods by Effrons while estimating pvalues where the hypothesis decision is based, the accuracy of the test was 100% successful. From the above results the reports showed that bootstrapping methods while testing or estimating some statistics should always considered because at most cases the outcome are accurate and errors are minimized in the computation. Also the RobustSharpe test which is known to use one of the bootstrapping methods, studentised one, were applied first on different simulated data including distribution of many kind and different shape secondly, on real data, Hedge and Mutual funds. The test performed quite well to agree with the existence of statistical significance difference between their Sharpe ratios as described in the paper of Ledoit andWolf.

Aging based maintenance and reinvestment scheduling of electric distribution

The maintenance of electric distribution network is a topical question for distribution system operators because of increasing significance of failure costs. In this dissertation the maintenance practices of the distribution system operators are analyzed and a theory for scheduling maintenance activities and reinvestment of distribution components is created. The scheduling is based on the deterioration of components and the increasing failure rates due to aging. The dynamic programming algorithm is used as a solving method to maintenance problem which is caused by the increasing failure rates of the network. The other impacts of network maintenance like environmental and regulation reasons are not included to the scope of this thesis. Further the tree trimming of the corridors and the major disturbance of the network are not included to the problem optimized in this thesis. For optimizing, four dynamic programming models are presented and the models are tested. Programming is made in VBA-language to the computer. For testing two different kinds of test networks are used. Because electric distribution system operators want to operate with bigger component groups, optimal timing for component groups is also analyzed. A maintenance software package is created to apply the presented theories in practice. An overview of the program is presented.

Testing Platform Development for Large Scale Solar Integration

Currently, a high penetration level of Distributed Generations (DGs) has been observed in the Danish distribution systems, and even more DGs are foreseen to be present in the upcoming years. How to utilize them for maintaining the security of the power supply under the emergency situations, has been of great interest for study. This master project is intended to develop a control architecture for studying purposes of distribution systems with large scale integration of solar power. As part of the EcoGrid EU Smart Grid project, it focuses on the system modelling and simulation of a Danish representative LV network located in Bornholm island. Regarding the control architecture, two types of reactive control techniques are implemented and compare. In addition, a network voltage control based on a tap changer transformer is tested. The optimized results after applying a genetic algorithm to five typical Danish domestic loads are lower power losses and voltage deviation using Q(U) control, specially with large consumptions. Finally, a communication and information exchange system is developed with the objective of regulating the reactive power and thereby, the network voltage remotely and real-time. Validation test of the simulated parameters are performed as well.

Development and evaluation of a loop-mediated isothermal amplification (LAMP) assay for rapid detection of Actinobacillus pleuropneumoniae based the dsbE-like gene

This paper reports on the development and validation of a loop-mediated isothermal amplification assay (LAMP) for the rapid and specific detection of Actinobacillus pleuropneumoniae (A. pleuropneumoniae). A set of six primers were designed derived from the dsbE-like gene of A.pleuropneumoniae and validate the assay using 9 A. pleuropneumoniae reference/field strains, 132 clinical isolates and 9 other pathogens. The results indicated that positive reactions were confirmed for all A. pleuropneumoniae strains and specimens by LAMP at 63ºC for 60 min and no cross-reactivity were observed from other non-A.pleuropneumoniae including Haemophilus parasuis, Escherichia coli, Pasteurella multocida, Bordetella bronchiseptica, Streptococcus suis, Salmonella enterica, Staphylococcus, porcine reproductive and respiratory syndrome virus (PRRSV), and Pseudorabies virus. The detection limit of the conventional PCR was 10² CFU per PCR test tube, while that of the LAMP was 5 copies per tube. Therefore, the sensitivity of LAMP was higher than that of PCR. Moreover, the LAMP assay provided a rapid yet simple test of A. pleuropneumoniae suitable for laboratory diagnosis and pen-side detection due to ease of operation and the requirement of only a regular water bath or heat block for the reaction.

Introduction of the T-10 static line parachuting capability to the NH90 helicopter

The purpose of this study was to investigate the suitability of the Finnish Defence Forces’ NH90 helicopter for parachuting operations with the T-10 static line parachute system. The work was based on the Army Command’s need to compensate for the reduction in the outsourced flight hours for the military static line parachuting training. The aim of the research was to find out the procedures and limitations with which the NH90 IOC+ or FOC version helicopter could be used for static line parachutist training with the T-10B/MC1-1C parachutes. The research area was highly complicated and non-linear. Thus analytical methods could not be applied with sufficient confidence, even with present-day computing power. Therefore an empirical research method was selected, concentrating on flight testing supported with literature study and some calculated estimations. During three flights and 4.5 flight hours in Utti, Finland on 1720 September 2012, a total of 44 parachute drops were made. These consisted of 16 dummy drops and 28 paratrooper jumps. The test results showed that when equipped with the floor mounted PASI-1 anchor line, the deflector bar of the NHIndustries’ Parachuting Kit and Patria’s floor protection panels the Finnish NH90 variant could be safely used for T-10B/MC1-1C static line parachuting operations from the right cabin door at airspeed range of 5080 KIAS (90–150 km/h). The ceiling mounted anchor lines of the NHI’s Parachuting Kit were not usable with the T-10 system. This was due to the static lines’ unsafe behaviour in slipstream when connected to the cabin ceiling level. In conclusion, the NH90 helicopter can be used to meet the Army Command’s requirement for an additional platform for T-10 static line parachutist training. Material dropping, the effect of additional equipment and jumping from the rear ramp should be further studied.

Testing pigs of non-technified rearing farms for serum antibodies against Taenia solium in a region of the state of São Paulo, Brazil

Abstract: Taenia solium is a zoonotic tapeworm of great importance in developing countries, due to the occurrence of human taeniasis and cysticercosis. Pigs have an important role in the biological cycle of the parasite as intermediate hosts. The scientific literature has been describing risk factors associated with the occurrence of this disease that must be avoided in countries with poor sanitation, in order to reduce the exposure of swine to the parasite eggs. This research focused on testing pigs of non-technified rearing farms for serum antibodies against Taenia solium in the region of Jaboticabal municipality, in the state of São Paulo, Brazil. The found prevalence was 6.82% (CI 95% 4.18 - 9.45) at animal level and 28.87% (CI 95% 16.74 - 40.40) at herd level. These figures are probably associated with low technification adoption during animal rearing in the studied area, which increased the exposure of the animals to risk factors associated with the occurrence of Taenia solium complex. The results found based on serological evidences of swine cysticercosis in the studied region serves as a warning to public sanitary authorities to improve public health and control T. solium.

Introduction of the T-10 static line parachuting capability to the NH90 helicopter

The purpose of this study was to investigate the suitability of the Finnish Defence Forces’ NH90 helicopter for parachuting operations with the T-10 static line parachute system. The work was based on the Army Command’s need to compensate for the reduction in the outsourced flight hours for the military static line parachuting training. The aim of the research was to find out the procedures and limitations with which the NH90 IOC+ or FOC version helicopter could be used for static line parachutist training with the T- 10B/MC1-1C parachutes. The research area was highly complicated and non-linear. Thus analytical methods could not be applied with sufficient confidence, even with present-day computing power. Therefore an empirical research method was selected, concentrating on flight testing supported with literature study and some calculated estimations. During three flights and 4.5 flight hours in Utti, Finland on 17−20 September 2012, a total of 44 parachute drops were made. These consisted of 16 dummy drops and 28 paratrooper jumps. The test results showed that when equipped with the floor mounted PASI-1 anchor line, the deflector bar of the NHIndustries’ Parachuting Kit and Patria’s floor protection panels the Finnish NH90 variant could be safely used for T-10B/MC1-1C static line parachuting operations from the right cabin door at airspeed range of 50−80 KIAS (∼90–150 km/h). The ceiling mounted anchor lines of the NHI’s Parachuting Kit were not usable with the T-10 system. This was due to the static lines’ unsafe behaviour in slipstream when connected to the cabin ceiling level. In conclusion, the NH90 helicopter can be used to meet the Army Command’s requirement for an additional platform for T-10 static line parachutist training. Material dropping, the effect of additional equipment and jumping from the rear ramp should be further studied.

Design and validation of stateful composite RESTful web services

A web service is a software system that provides a machine-processable interface to the other machines over the network using different Internet protocols. They are being increasingly used in the industry in order to automate different tasks and offer services to a wider audience. The REST architectural style aims at producing scalable and extensible web services using technologies that play well with the existing tools and infrastructure of the web. It provides a uniform set of operation that can be used to invoke a CRUD interface (create, retrieve, update and delete) of a web service. The stateless behavior of the service interface requires that every request to a resource is independent of the previous ones facilitating scalability. Automated systems, e.g., hotel reservation systems, provide advanced scenarios for stateful services that require a certain sequence of requests that must be followed in order to fulfill the service goals. Designing and developing such services for advanced scenarios with REST constraints require rigorous approaches that are capable of creating web services that can be trusted for their behavior. Systems that can be trusted for their behavior can be termed as dependable systems. This thesis presents an integrated design, analysis and validation approach that facilitates the service developer to create dependable and stateful REST web services. The main contribution of this thesis is that we provide a novel model-driven methodology to design behavioral REST web service interfaces and their compositions. The behavioral interfaces provide information on what methods can be invoked on a service and the pre- and post-conditions of these methods. The methodology uses Unified Modeling Language (UML), as the modeling language, which has a wide user base and has mature tools that are continuously evolving. We have used UML class diagram and UML state machine diagram with additional design constraints to provide resource and behavioral models, respectively, for designing REST web service interfaces. These service design models serve as a specification document and the information presented in them have manifold applications. The service design models also contain information about the time and domain requirements of the service that can help in requirement traceability which is an important part of our approach. Requirement traceability helps in capturing faults in the design models and other elements of software development environment by tracing back and forth the unfulfilled requirements of the service. The information about service actors is also included in the design models which is required for authenticating the service requests by authorized actors since not all types of users have access to all the resources. In addition, following our design approach, the service developer can ensure that the designed web service interfaces will be REST compliant. The second contribution of this thesis is consistency analysis of the behavioral REST interfaces. To overcome the inconsistency problem and design errors in our service models, we have used semantic technologies. The REST interfaces are represented in web ontology language, OWL2, that can be part of the semantic web. These interfaces are used with OWL 2 reasoners to check unsatisfiable concepts which result in implementations that fail. This work is fully automated thanks to the implemented translation tool and the existing OWL 2 reasoners. The third contribution of this thesis is the verification and validation of REST web services. We have used model checking techniques with UPPAAL model checker for this purpose. The timed automata of UML based service design models are generated with our transformation tool that are verified for their basic characteristics like deadlock freedom, liveness, reachability and safety. The implementation of a web service is tested using a black-box testing approach. Test cases are generated from the UPPAAL timed automata and using the online testing tool, UPPAAL TRON, the service implementation is validated at runtime against its specifications. Requirement traceability is also addressed in our validation approach with which we can see what service goals are met and trace back the unfulfilled service goals to detect the faults in the design models. A final contribution of the thesis is an implementation of behavioral REST interfaces and service monitors from the service design models. The partial code generation tool creates code skeletons of REST web services with method pre and post-conditions. The preconditions of methods constrain the user to invoke the stateful REST service under the right conditions and the post condition constraint the service developer to implement the right functionality. The details of the methods can be manually inserted by the developer as required. We do not target complete automation because we focus only on the interface aspects of the web service. The applicability of the approach is demonstrated with a pedagogical example of a hotel room booking service and a relatively complex worked example of holiday booking service taken from the industrial context. The former example presents a simple explanation of the approach and the later worked example shows how stateful and timed web services offering complex scenarios and involving other web services can be constructed using our approach.

Analyses of the usefulness of Software Defined Storage Solutions for Web-based Digital Preservation Applications

Presentation at Open Repositories 2014, Helsinki, Finland, June 9-13, 2014

Modal testing and numerical modeling of the dynamic properties of layered sheet-steel structure

Recently, due to the increasing total construction and transportation cost and difficulties associated with handling massive structural components or assemblies, there has been increasing financial pressure to reduce structural weight. Furthermore, advances in material technology coupled with continuing advances in design tools and techniques have encouraged engineers to vary and combine materials, offering new opportunities to reduce the weight of mechanical structures. These new lower mass systems, however, are more susceptible to inherent imbalances, a weakness that can result in higher shock and harmonic resonances which leads to poor structural dynamic performances. The objective of this thesis is the modeling of layered sheet steel elements, to accurately predict dynamic performance. During the development of the layered sheet steel model, the numerical modeling approach, the Finite Element Analysis and the Experimental Modal Analysis are applied in building a modal model of the layered sheet steel elements. Furthermore, in view of getting a better understanding of the dynamic behavior of layered sheet steel, several binding methods have been studied to understand and demonstrate how a binding method affects the dynamic behavior of layered sheet steel elements when compared to single homogeneous steel plate. Based on the developed layered sheet steel model, the dynamic behavior of a lightweight wheel structure to be used as the structure for the stator of an outer rotor Direct-Drive Permanent Magnet Synchronous Generator designed for high-power wind turbines is studied.