Microclimate and gas emissions in dairy buildings : Instrumentation, theory and measurements

The aim of this thesis was to develop measurement techniques and systems for measuring air quality and to provide information about air quality conditions and the amount of gaseous emissions from semi-insulated and uninsulated dairy buildings in Finland and Estonia. Specialization and intensification in livestock farming, such as in dairy production, is usually accompanied by an increase in concentrated environmental emissions. In addition to high moisture, the presence of dust and corrosive gases, and widely varying gas concentrations in dairy buildings, Finland and Estonia experience winter temperatures reaching below -40 ºC and summer temperatures above +30 ºC. The adaptation of new technologies for long-term air quality monitoring and measurement remains relatively uncommon in dairy buildings because the construction and maintenance of accurate monitoring systems for long-term use are too expensive for the average dairy farmer to afford. Though the documentation of accurate air quality measurement systems intended mainly for research purposes have been made in the past, standardised methods and the documentation of affordable systems and simple methods for performing air quality and emissions measurements in dairy buildings are unavailable. In this study, we built three measurement systems: 1) a Stationary system with integrated affordable sensors for on-site measurements, 2) a Wireless system with affordable sensors for off-site measurements, and 3) a Mobile system consisting of expensive and accurate sensors for measuring air quality. In addition to assessing existing methods, we developed simplified methods for measuring ventilation and emission rates in dairy buildings. The three measurement systems were successfully used to measure air quality in uninsulated, semi-insulated, and fully-insulated dairy buildings between the years 2005 and 2007. When carefully calibrated, the affordable sensors in the systems gave reasonably accurate readings. The spatial air quality survey showed high variation in microclimate conditions in the dairy buildings measured. The average indoor air concentration for carbon dioxide was 950 ppm, for ammonia 5 ppm, for methane 48 ppm, for relative humidity 70%, and for inside air velocity 0.2 m/s. The average winter and summer indoor temperatures during the measurement period were -7º C and +24 ºC for the uninsulated, +3 ºC and +20 ºC for the semi-insulated and +10 ºC and +25 ºC for the fully-insulated dairy buildings. The measurement results showed that the uninsulated dairy buildings had lower indoor gas concentrations and emissions compared to fully insulated buildings. Although occasionally exceeded, the ventilation rates and average indoor air quality in the dairy buildings were largely within recommended limits. We assessed the traditional heat balance, moisture balance, carbon dioxide balance and direct airflow methods for estimating ventilation rates. The direct velocity measurement for the estimation of ventilation rate proved to be impractical for naturally ventilated buildings. Two methods were developed for estimating ventilation rates. The first method is applicable in buildings in which the ventilation can be stopped or completely closed. The second method is useful in naturally ventilated buildings with large openings and high ventilation rates where spatial gas concentrations are heterogeneously distributed. The two traditional methods (carbon dioxide and methane balances), and two newly developed methods (theoretical modelling using Fick s law and boundary layer theory, and the recirculation flux-chamber technique) were used to estimate ammonia emissions from the dairy buildings. Using the traditional carbon dioxide balance method, ammonia emissions per cow from the dairy buildings ranged from 7 g day-1 to 35 g day-1, and methane emissions per cow ranged from 96 g day-1 to 348 g day-1. The developed methods proved to be as equally accurate as the traditional methods. Variation between the mean emissions estimated with the traditional and the developed methods was less than 20%. The developed modelling procedure provided sound framework for examining the impact of production systems on ammonia emissions in dairy buildings.

The Pseudomonas syringae-derived HrpA pilins - molecular characterization and biotechnological application of the transcripts

Many Gram-negative bacteria pathogenic to plants and animals possess type III secretion systems that are used to cause disease. Effector proteins are injected into host cells using the type III secretion machineries. Despite vigorous studies, the nature of the secretion signal for type III secreted proteins still remains elusive. Both mRNA and proteinaceous signals have been proposed. Findings on coupling of translation to secretion by the type III secretion systems are also still contradictory. This study dealt with the secretion signal of HrpA from Pseudomonas syringae pathovar tomato. HrpA is the major component of the type III secretion system-associated Hrp pilus and a substrate for the type III secretion systems. The secretion signal was shown to reside in the first 15 codons or amino acids, a location typical for type III secretion signals. Translation of HrpA in the absence of a functional type III secretion system was established, but it does not exclude the possibility of coupling of translation to secretion when the secretion apparatus is present. The hrpA transcripts from various unrelated plant pathogenic bacteria were shown to be extremely stable. The biological relevance of this observation is unknown, but possible explanations include the high prevalence of HrpA protein, an mRNA secretion signal or timing of secretion. The hrpA mRNAs are stable over a wide range of temperatures, in the absence of translating ribosomes and even in the heterologous host Escherichia coli. The untranslated regions (UTRs) of hrpA transcripts from at least 20 pathovars of Pseudomonas syringae are highly homologous, whilst their coding regions exhibit low similarity. The stable nature of hrpA messenger RNAs is likely to be due to the folding of their 5 and 3 UTRs. In silico the UTRs seem to form stem-loop structures, the hairpin structures in the 3 UTRs being rich in guanidine and cytosine residues. The stable nature of the hrpA transcript redirected the studies to the stabilization of heterologous transcripts and to the use of stable messenger RNAs in recombinant protein production. Fragments of the hrpA transcript can be used to confer stability on heterologous transcripts from several sources of bacterial and eukaryotic origin, and to elevate the levels of production of the corresponding recombinant proteins several folds. hrpA transcript stabilizing elements can be used for improving the yields of recombinant proteins even in Escherichia coli, one of the most commonly used industrial protein production hosts.

New Method for Delexicalization and its Application to Prosodic Tagging for Text-to-Speech Synthesis

This paper describes a new flexible delexicalization method based on glottal excited parametric speech synthesis scheme. The system utilizes inverse filtered glottal flow and all-pole modelling of the vocal tract. The method provides a possibil- ity to retain and manipulate all relevant prosodic features of any kind of speech. Most importantly, the features include voice quality, which has not been properly modeled in earlier delex- icalization methods. The functionality of the new method was tested in a prosodic tagging experiment aimed at providing word prominence data for a text-to-speech synthesis system. The ex- periment confirmed the usefulness of the method and further corroborated earlier evidence that linguistic factors influence the perception of prosodic prominence.