Bacillus cereus spores and cereulide in food-borne illness

B. cereus is a gram-positive bacterium that possesses two different forms of life:the large, rod-shaped cells (ca. 0.002 mm by 0.004 mm) that are able to propagate and the small (0.001 mm), oval shaped spores. The spores can survive in almost any environment for up to centuries without nourishment or water. They are insensitive towards most agents that normally kill bacteria: heating up to several hours at 90 ºC, radiation, disinfectants and extreme alkaline (≥ pH 13) and acid (≤ pH 1) environment. The spores are highly hydrophobic and therefore make them tend to stick to all kinds of surfaces, steel, plastics and live cells. In favorable conditions the spores of B. cereus may germinate into vegetative cells capable of producing food poisoning toxins. The toxins can be heat-labile protein formed after ingestion of the contaminated food, inside the gastrointestinal tract (diarrhoeal toxins), or heat stable peptides formed in the food (emesis causing toxin, cereulide). Cereulide cannot be inactivated in foods by cooking or any other procedure applicable on food. Cereulide in consumed food causes serious illness in human, even fatalities. In this thesis, B. cereus strains originating from different kinds of foods and environments and 8 different countries were inspected for their capability of forming cereulide. Of the 1041 isolates from soil, animal feed, water, air, used bedding, grass, dung and equipment only 1.2 % were capable of producing cereulide, whereas of the 144 isolates originating from foods 24 % were cereulide producers. Cereulide was detected by two methods: by its toxicity towards mammalian cells (sperm assay) and by its peculiar chemical structure using liquid-chromatograph-mass spectrometry equipment. B. cereus is known as one of the most frequent bacteria occurring in food. Most foods contain more than one kind of B. cereus. When randomly selected 100 isolates of B. cereus from commercial infant foods (dry formulas) were tested, 11% of these produced cereulide. Considering a frequent content of 103 to 104 cfu (colony forming units) of B. cereus per gram of infant food formula (dry), it appears likely that most servings (200 ml, 30 g of the powder reconstituted with water) may contain cereulide producers. When a reconstituted infant formula was inoculated with >105 cfu of cereulide producing B. cereus per ml and left at room temperature, cereulide accumulated to food poisoning levels (> 0.1 mg of cereulide per serving) within 24 hours. Paradoxically, the amount of cereulide (per g of food) increased 10 to 50 fold when the food was diluted 4 - 15 fold with water. The amount of the produced cereulide strongly depended on the composition of the formula: most toxin was formed in formulas with cereals mixed with milk, and least toxin in formulas based on milk only. In spite of the aggressive cleaning practices executed by the modern dairy industry, certain genotypes of B. cereus appear to colonise the silos tanks. In this thesis four strategies to explain their survival of their spores in dairy silos were identified. First, high survival (log 15 min kill ≤ 1.5) in the hot alkaline (pH >13) wash liquid, used at the dairies for cleaning-in-place. Second, efficient adherence of the spores to stainless steel from cold water. Third, a cereulide producing group with spores characterized by slow germination in rich medium and well preserved viability when exposed to heating at 90 ºC. Fourth, spores capable of germinating at 8 ºC and possessing the psychrotolerance gene, cspA. There were indications that spores highly resistant to hot 1% sodium hydroxide may be effectively inactivated by hot 0.9% nitric acid. Eight out of the 14 dairy silo tank isolates possessing hot alkali resistant spores were capable of germinating and forming biofilm in whole milk, not previously reported for B. cereus. In this thesis it was shown that cereulide producing B. cereus was capable of inhibiting the growth of cereulide non-producing B. cereus occurring in the same food. This phenomenon, called antagonism, has long been known to exist between B. cereus and other microbial species, e.g. various species of Bacillus, gram-negative bacteria and plant pathogenic fungi. In this thesis intra-species antagonism of B. cereus was shown for the first time. This brother-killing did not depend on the cereulide molecule, also some of the cereulide non-producers were potent antagonists. Interestingly, the antagonistic clades were most frequently found in isolates from food implicated with human illness. The antagonistic property was therefore proposed in this thesis as a novel virulence factor that increases the human morbidity of the species B. cereus, in particular of the cereulide producers.

Drops That Open Worlds : Image of Water in the Poetry of Euphrase Kezilahabi

Euphrase Kezilahabi on tansanialainen kirjailija, joka ensimmäisenä julkaisi swahilinkielisen vapaalla mitalla kirjoitetun runokokoelman. Perinteisessä swahilirunoudessa tiukat muotosäännöt ovat tärkeitä, ja teos synnytti kiivasta keskustelua. Runoteokset Kichomi ( Viilto , Kipu , 1974) ja Karibu Ndani ( Tervetuloa sisään , 1988) sekä Kezilahabin muu tuotanto voidaan nähdä uuden sukupolven taiteena. Kezilahabi on arvostettu runoilija, mutta hänen runojaan ei aiemmin ole käännetty englanniksi (yksittäisiä säkeitä lukuunottamatta), eikä juurikaan tutkittu yksityiskohtaisesti. Yleiskuvaan pyrkivissä lausunnoissa Kezilahabin runouden on hyvin usein määritelty olevan poliittista. Monet Kezilahabin runoista ottavatkin kantaa yhteiskunnallisiin kysymyksiin, mutta niiden pohdinta on kuitenkin runoissa vain yksi taso. Sen lisäksi Kezilahabin lyriikassa on paljon muuta ennen kartoittamatonta tämä tutkimus keskittyy veden kuvaan (the image of water). Kezilahabi vietti lapsuutensa saarella Victoria-järven keskellä, ja hänen vesikuvastonsa on rikasta. Tutkimuskysymyksenä on, mitä veden kuva runoteoksissa Kichomi ja Karibu Ndani esittää. Runojen analysoinnissa ja tulkinnassa on tarkasteltu myös sitä, miten äänteellinen taso osallistuu kuvien luomiseen. Tutkimuksen määritelmä kuvasta pohjautuu osittain Hugh Kennerin näkemykseen, jonka mukaan oleellista kuvassa on kirjaimellinen taso. Kennerin lähtökohtaan on yhdistetty John Shoptawin teoriaa, joka korostaa runon äänteellisen puolen tärkeyttä merkityksen muodostumisessa. Foneemien analyysissä vaikutteena on ollut Reuven Tsurin teoria. Analyysiosio osoittaa, että veden kuva edustaa ja käsittelee teoksissa lukuisia teemoja: elämää, kuolemaa, fyysistä vetovoimaa, runoutta, mielikuvitusta ja (ali)tajuntaa sekä moraalia. Veden kuvan tutkimuksen pohjalta on nähtävissä, että Kezilahabin filosofia asettuu elävä/kuollut- ja elämä/kuolema dikotomioiden ulkopuolelle.

Rearing strategies of young dairy calves in relation to production, behaviour and welfare

Vasikoiden kasvatus yksilökarsinoissa, imemismahdollisuuden puute maitojuoton yhteydessä sekä pienet juomamäärät ovat tekijöitä, jotka mahdollisesti voivat vähentää vasikoiden hyvinvointia. Vasikoiden kasvatukseen etsitäänkin uusia tapoja, joissa eläinten käyttäytymistarpeet ja hyvinvointi otetaan entistä paremmin huomioon. Tässä väitöskirjassa vasikoiden kasvatusta on tarkasteltu sekä tuotannon että vasikoiden käyttäytymisen ja hyvinvoinnin kannalta. Väitöskirja koostuu kolmesta kokeesta, joista ensimmäisessä tutkittiin vasikoiden kasvatusta ryhmäkarsinoissa ulkona tai sisällä ja vasikoiden kasvatusta sisällä ryhmä- tai yksilökarsinoissa. Toisessa kokeessa vasikoiden annettiin imeä emiään rajoitetusti lypsyn jälkeen viiden tai kahdeksan viikon ajan ja kolmannessa selvitettiin vasikoiden veden juontia, kun vasikat saivat juomarehua vapaasti. Lisäksi kokeiden yhdistetystä aineistosta analysoitiin eri rehujen syöntimäärien suhdetta sekä rehujen vaikutusta kasvuun ennen ja jälkeen maidosta vieroituksen. Tutkimuksessa todettiin, että vasikoita voi ryhmässä kasvattaa kylmissä ja vaihtelevissa sääoloissa ulkona, kunhan ne hoidetaan ja ruokitaan erittäin huolellisesti. Kylmällä ilmalla vasikat saattavat kuitenkin syödä väkirehua vähemmän varsinkin, jos ruokailupaikka on ulkona ja makuualue sisällä. Ryhmässä kasvaneet vasikat aloittivat sekä kuivien rehujen syönnin että märehtimisen nuorempina kuin yksilökarsinassa kasvaneet. Ryhmissä esiintyvää käyttäytymisongelmaa, toisten vasikoiden imemistä, voidaan vähentää hoito- ja ruokintamenetelmillä. Annettaessa vasikoiden imeä emiään rajoitetusti lypsyn jälkeen vasikat oppivat imemään emiään hyvin nopeasti. Lypsytyö vaikeutui muutamien lehmien kohdalla, sillä ne pidättivät maitoa lypsettäessä. Saadessaan imeä rajoitetusti vasikat imivät suurehkoja maitomääriä kerrallaan. Vieroittaminen suurilta maitomääriltä viiden viikon iässä oli kuitenkin liian aikaista, koska vasikat eivät vielä syöneet riittävästi kuivia rehuja. Vieroitus emästä niin viiden kuin kahdeksankin viikon iässä aiheutti vasikoissa levottomuutta ja ääntelyn lisääntymistä. Saadessaan hapatettua juomarehua vapaasti vasikat joivat keskimäärin vain vähän vettä, olipa vesilähteenä avoin ämpäri tai vesinippa. Vasikoiden välillä oli suurta vaihtelua veden juontimäärissä. Viikkoa ennen maidosta vieroitusta vasikat joivat 0-3 l vettä päivässä. Vasikat joivat nipasta kerrallaan vähemmän vettä kuin ämpäristä, ja käyttivät enemmän aikaa päivässä veden juomiseen kuin vesiämpäristä juoneet vasikat. Suurin osa vasikoista joi vettä juomanipoista erikoisella tavalla esimerkiksi painamalla nippaa otsalla ja juomalla tippuvaa vettä. Vesinipat voivat olla siis vasikoille joko vaikeita tai epämukavia käyttää. Vasikoiden juoman maitomäärän lisääntyessä kasvu lisääntyy selvästi. Runsas maidon juominen vähentää kuitenkin kuivien rehujen syöntiä ja vieroitusvaiheessa kasvu voi hidastua. Vasikat olisikin tärkeää vieroittaa vähitellen, ettei muutoksia kasvuun tulisi. Syönti- ja kasvutulokset eivät aina anna oikeaa kuvaa kasvatusmenetelmien eroista eläinten hyvinvoinnin kannalta. Käyttäytyminen on herkkä hyvinvoinnin mittari ja se tulisikin aina huomioida eri kasvatusmenetelmiä arvioitaessa. .

Winter feeding strategies for suckler cows in cold climatic conditions

In Finland, suckler cow production is carried out in circumstances characterized by a long winter period and a short grazing period. The traditional winter housing system for suckler cows has been insulated or uninsulated buildings, but there is a demand for developing less expensive housing systems. In addition, more information is needed on new winter feeding strategies, carried out in inexpensive winter facilities with conventional (hay, grass silage, straw) or alternative (treated straw, industrial by-product, whole-crop silage) feeds. The new feeding techniques should not have any detrimental effects on animal welfare in order to be acceptable to both farmers and consumers. Furthermore, no official feeding recommendations for suckler cows are available in Finland and, thus, recommendations for dairy cows have been used. However, this may lead to over- or underfeeding of suckler cows and, finally, to decreased economic output. In Experiment I, second-calf beef-dairy suckler cows were used to compare the effects of diets based on hay (H) or urea-treated straw (US) at two feeding levels (Moderate; M vs. Low; L) on the performance of cows and calves. Live weight (LW) gain during the indoor feeding was lower for cows on level L than on level M. Cows on diet US lost more LW indoors than those on diet H. The cows replenished the LW losses on good pasture. Calf LW gain and cow milk production were unaffected by the treatments. Conception rate was unaffected by the treatments but was only 69%. Urea-treated straw proved to be a suitable winter feed for spring-calving suckler cows. Experiment II studied the effects of feeding accuracy on the performance of first- and second-calf beef-dairy cows and calves. In II-1, the day-to-day variation in the roughage offered ranged up to ± 40%. In II-2, the same variation was used in two-week periods. Variation of the roughages offered had minor effects on cow performance. Reproduction was unaffected by the feeding accuracy. Accurate feeding is not necessary for young beef-dairy crosses, if the total amount of energy offered over a period of a few weeks fulfills the energy requirements. Effects of feeding strategies with alternative feeds on the performance of mature beef-dairy and beef cows and calves were evaluated in Experiment III. Two studies consisted of two feeding strategies (Step-up vs. Flat-rate) and two diets (Control vs. Alternative). There were no differences between treatments in the cow LW, body condition score (BCS), calf pre-weaning LW gain and cow reproduction. A flat-rate strategy can be practised in the nutrition of mature suckler cows. Oat hull based flour-mill by product can partly replace grass silage and straw in the winter diet. Whole-crop barley silage can be offered as a sole feed to suckler cows. Experiment IV evaluated during the winter feeding period the effects of replacing grass silage with whole-crop barley or oat silage on mature beef cow and calf performance. Both whole-crop silages were suitable winter feeds for suckler cows in cold outdoor winter conditions. Experiment V aimed at assessing the effects of daily feeding vs. feeding every third day on the performance of mature beef cows and calves. No differences between the treatments were observed in cow LW, BCS, milk production and calf LW. The serum concentrations of urea and long-chain fatty acids were increased on the third day after feeding in the cows fed every third day. Despite of that the feeding every third day is an acceptable feeding strategy for mature suckler cows. Experiment VI studied the effects of feeding levels and long-term cold climatic conditions on mature beef cows and calves. The cows were overwintered in outdoor facilities or in an uninsulated indoor facility. Whole-crop barley silage was offered either ad libitum or restricted. All the facilities offered adequate shelter for the cows. The restricted offering of whole-crop barley silage provided enough energy for the cows. The Finnish energy recommendations for dairy cows were too high for mature beef breed suckler cows in good body condition at housing, even in cold conditions. Therefore, there is need to determine feeding recommendations for suckler cows in Finland. The results showed that the required amount of energy can be offered to the cows using conventional or alternative feeds provided at a lower feeding level, with an inaccurate feeding, flat-rate feeding or feeding every third day strategy. The cows must have an opportunity to replenish the LW and BCS losses at pasture before the next winter. Production in cold conditions can be practised in inexpensive facilities when shelter against rain and wind, a dry resting place, adequate amounts of feed suitable for cold conditions and water are provided for the animals as was done in the present study.

Heterotrophic Bacteria Associated with Cyanobacteria in Recreational and Drinking Water

Cyanobacterial mass occurrences, also known as water blooms, have been associated with adverse health effects of both humans and animals. They can also be a burden to drinking water treatment facilities. Risk assessments of the blooms have generally focused on the cyanobacteria themselves and their toxins. However, heterotrophic bacteria thriving among cyanobacteria may also be responsible for many of the adverse health effects, but their role as the etiological agents of these health problems is poorly known. In addition, studies on the water purification efficiency of operating water treatment plants during cyanobacterial mass occurrences in their water sources are rare. In the present study, over 600 heterotrophic bacterial strains were isolated from natural freshwater, brackish water or from treated drinking water. The sampling sites were selected as having frequent cyanobacterial occurrences in the water bodies or in the water sources of the drinking water treatment plants. In addition, samples were taken from sites where cyanobacterial water blooms were surmised to have caused human health problems. The isolated strains represented bacteria from 57 different genera of the Gamma-, Alpha- or Betaproteobacteria, Actinobacteria, Flavobacteria, Sphingobacteria, Bacilli and Deinococci classes, based on their partial 16S rRNA sequences. Several isolates had no close relatives among previously isolated bacteria or cloned 16S rRNA genes of uncultivated bacteria. The results show that water blooms are associated with a diverse community of cultivable heterotrophic bacteria. Chosen subsets of the isolated strains were analysed for features such as their virulence gene content and possible effect on cyanobacterial growth. Of the putatively pathogenic haemolytic strains isolated in the study, the majority represented the genus Aeromonas. Therefore, the Aeromonas spp. strains isolated from water samples associated with adverse health effects were screened for the virulence gene types encoding for enterotoxins (ast, alt and act/aerA/hlyA), flagellin subunits (flaA/flaB), lipase (lip/pla/lipH3/alp-1) and elastase (ahyB) by PCR. The majority (90%) of the Aeromonas strains included one or more of the six screened Aeromonas virulence gene types. The most common gene type was act, which was present in 77% of the strains. The fla, ahyB and lip genes were present in 30 37% of the strains. The prevalence of the virulence genes implies that the Aeromonas may be a factor in some of the cyanobacterial associated health problems. Of the 183 isolated bacterial strains that were studied for possible effects on cyanobacterial growth, the majority (60%) either enhanced or inhibited growth of cyanobacteria. In most cases, they enhanced the growth, which implies mutualistic interactions. The results indicate that the heterotrophic bacteria have a role in the rise and fall of the cyanobacterial water blooms. The genetic and phenotypic characteristics and the ability to degrade cyanobacterial hepatotoxins of 13 previously isolated Betaproteobacteria strains, were also studied. The strains originated from Finnish lakes with frequent cyanobacterial occurrence. Tested strains degraded microcystins -LR and -YR and nodularin. The strains could not be assigned to any described bacterial genus or species based on their genetic or phenotypic features. On the basis of their characteristics a new genus and species Paucibacter toxinivorans was proposed for them. The water purification efficiency of the drinking water treatment processes during cyanobacterial water bloom in water source was assessed at an operating surface water treatment plant. Large phytoplankton, cyanobacterial hepatotoxins, endotoxins and cultivable heterotrophic bacteria were efficiently reduced to low concentrations, often below the detection limits. In contrast, small planktonic cells, including also possible bacterial cells, regularly passed though the water treatment. The passing cells may contribute to biofilm formation within the water distribution system, and therefore lower the obtained drinking water quality. The bacterial strains of this study offer a rich source of isolated strains for examining interactions between cyanobacteria and the heterotrophic bacteria associated with them. The degraders of cyanobacterial hepatotoxins could perhaps be utilized to assist the removal of the hepatotoxins during water treatment, whereas inhibitors of cyanobacterial growth might be useful in controlling cyanobacterial water blooms. The putative pathogenicity of the strains suggests that the health risk assessment of the cyanobacterial blooms should also cover the heterotrophic bacteria.

Water-ethanol mixtures by molecular dynamics and x-ray Compton scattering

Water-ethanol mixtures are commonly used in industry and house holds. However, quite surprisingly their molecular-level structure is still not completely understood. In particular, there is evidence that the local intermolecular geometries depend significantly on the concentration. The aim of this study was to gain information on the molecular-level structures of water-ethanol mixtures by two computational methods. The methods are classical molecular dynamics (MD), where the movement of molecules can be studied, and x-ray Compton scattering, in which the scattering cross section is sensitive to the electron momentum density. Firstly, the water-ethanol mixtures were studied with MD simulations, with the mixture concentration ranging from 0 to 100%. For the simulations well-established force fields were used for the water and ethanol molecules (TIP4P and OPLS-AA, respectively). Moreover, two models were used for ethanol, rigid and non-rigid. In the rigid model the intramolecular bond lengths are fixed, whereas in the non-rigid model the lengths are determined by harmonic potentials. Secondly, mixtures with three different concentrations employing both ethanol models were studied by calculating the experimentally observable x-ray quantity, the Compton profile. In the MD simulations a slight underestimation in the density was observed as compared to experiment. Furthermore, a positive excess of hydrogen bonding with water molecules and a negative one with ethanol was quantified. Also, the mixture was found more structured when the ethanol concentration was higher. Negligible differences in the results were found between the two ethanol models. In contrast, in the Compton scattering results a notable difference between the ethanol models was observed. For the rigid model the Compton profiles were similar for all the concentrations, but for the non-rigid model they were distinct. This leads to two possibilities of how the mixing occurs. Either the mixing is similar in all concentrations (as suggested by the rigid model) or the mixing changes for different concentrations (as suggested by the non-rigid model). Either way, this study shows that the choice of the force field is essential in the microscopic structure formation in the MD simulations. When the sources of uncertainty in the calculated Compton profiles were analyzed, it was found that more statistics needs to be collected to reduce the statistical uncertainty in the final results. The obtained Compton scattering results can be considered somewhat preliminary, but clearly indicative of the behaviour of the water-ethanol mixtures when the force field is modified. The next step is to collect more statistics and compare the results with experimental data to decide which ethanol model describes the mixture better. This way, valuable information on the microscopic structure of water-ethanol mixtures can be found. In addition, information on the force fields in the MD simulations and on the ability of the MD simulations to reproduce the microscopic structure of binary liquids is obtained.

A computational study of Compton profiles of water - methanol solutions

The molecular level structure of mixtures of water and alcohols is very complicated and has been under intense research in the recent past. Both experimental and computational methods have been used in the studies. One method for studying the intra- and intermolecular bindings in the mixtures is the use of the so called difference Compton profiles, which are a way to obtain information about changes in the electron wave functions. In the process of Compton scattering a photon scatters inelastically from an electron. The Compton profile that is obtained from the electron wave functions is directly proportional to the probability of photon scattering at a given energy to a given solid angle. In this work we develop a method to compute Compton profiles numerically for mixtures of liquids. In order to obtain the electronic wave functions necessary to calculate the Compton profiles we need some statistical information about atomic coordinates. Acquiring this using ab-initio molecular dynamics is beyond our computational capabilities and therefore we use classical molecular dynamics to model the movement of atoms in the mixture. We discuss the validity of the chosen method in view of the results obtained from the simulations. There are some difficulties in using classical molecular dynamics for the quantum mechanical calculations, but these can possibly be overcome by parameter tuning. According to the calculations clear differences can be seen in the Compton profiles of different mixtures. This prediction needs to be tested in experiments in order to find out whether the approximations made are valid.

Asymmetrical flow field-flow fractionation in the study of water-soluble macromolecules

Asymmetrical flow field-flow fractionation (AsFlFFF) was constructed, and its applicability to industrial, biochemical, and pharmaceutical applications was studied. The effect of several parameters, such as pH, ionic strength, temperature and the reactants mixing ratios on the particle sizes, molar masses, and the formation of aggregates of macromolecules was determined by AsFlFFF. In the case of industrial application AsFlFFF proved to be a valuable tool in the characterization of the hydrodynamic particle sizes, molar masses and phase transition behavior of various poly(N-isopropylacrylamide) (PNIPAM) polymers as a function of viscosity and phase transition temperatures. The effect of sodium chloride salt and the molar ratio of cationic and anionic polyelectrolytes on the hydrodynamic particle sizes of poly (methacryloxyethyl trimethylammonium chloride) and poly (ethylene oxide)-block-poly (sodium methacrylate) and their complexes were studied. The particle sizes of PNIPAM polymers, and polyelectrolyte complexes measured by AsFlFFF were in agreement with those obtained by dynamic light scattering. The molar masses of PNIPAM polymers obtained by AsFlFFF and size exclusion chromatography agreed also well. In addition, AsFlFFF proved to be a practical technique in thermo responsive behavior studies of polymers at temperatures up to about 50 oC. The suitability of AsFlFFF for biological, biomedical, and pharmaceutical applications was proved, upon studying the lipid-protein/peptide interactions, and the stability of liposomes at different temperatures. AsFlFFF was applied to the studies on the hydrophobic and electrostatic interactions between cytochrome c (a basic peripheral protein) and anionic lipid, and oleic acid, and sodium dodecyl sulphate surfactant. A miniaturized AsFlFFF constructed in this study was exploited in the elucidation of the effect of copper (II), pH, ionic strength, and vortexing on the particle sizes of low-density lipoproteins.

Parameters Affecting the Extraction of Polycyclic Aromatic Hydrocarbons with Pressurised Hot Water

Pressurised hot water extraction (PHWE) exploits the unique temperature-dependent solvent properties of water minimising the use of harmful organic solvents. Water is environmentally friendly, cheap and easily available extraction medium. The effects of temperature, pressure and extraction time in PHWE have often been studied, but here the emphasis was on other parameters important for the extraction, most notably the dimensions of the extraction vessel and the stability and solubility of the analytes to be extracted. Non-linear data analysis and self-organising maps were employed in the data analysis to obtain correlations between the parameters studied, recoveries and relative errors. First, pressurised hot water extraction (PHWE) was combined on-line with liquid chromatography-gas chromatography (LC-GC), and the system was applied to the extraction and analysis of polycyclic aromatic hydrocarbons (PAHs) in sediment. The method is of superior sensitivity compared with the traditional methods, and only a small 10 mg sample was required for analysis. The commercial extraction vessels were replaced by laboratory-made stainless steel vessels because of some problems that arose. The performance of the laboratory-made vessels was comparable to that of the commercial ones. In an investigation of the effect of thermal desorption in PHWE, it was found that at lower temperatures (200ºC and 250ºC) the effect of thermal desorption is smaller than the effect of the solvating property of hot water. At 300ºC, however, thermal desorption is the main mechanism. The effect of the geometry of the extraction vessel on recoveries was studied with five specially constructed extraction vessels. In addition to the extraction vessel geometry, the sediment packing style and the direction of water flow through the vessel were investigated. The geometry of the vessel was found to have only minor effect on the recoveries, and the same was true of the sediment packing style and the direction of water flow through the vessel. These are good results because these parameters do not have to be carefully optimised before the start of extractions. Liquid-liquid extraction (LLE) and solid-phase extraction (SPE) were compared as trapping techniques for PHWE. LLE was more robust than SPE and it provided better recoveries and repeatabilities than did SPE. Problems related to blocking of the Tenax trap and unrepeatable trapping of the analytes were encountered in SPE. Thus, although LLE is more labour intensive, it can be recommended over SPE. The stabilities of the PAHs in aqueous solutions were measured using a batch-type reaction vessel. Degradation was observed at 300ºC even with the shortest heating time. Ketones and quinones and other oxidation products were observed. Although the conditions of the stability studies differed considerably from the extraction conditions in PHWE, the results indicate that the risk of analyte degradation must be taken into account in PHWE. The aqueous solubilities of acenaphthene, anthracene and pyrene were measured, first below and then above the melting point of the analytes. Measurements below the melting point were made to check that the equipment was working, and the results were compared with those obtained earlier. Good agreement was found between the measured and literature values. A new saturation cell was constructed for the solubility measurements above the melting point of the analytes because the flow-through saturation cell could not be used above the melting point. An exponential relationship was found between the solubilities measured for pyrene and anthracene and temperature.

Precipitation-induced runoff and leaching from milled peat mining mires by peat types : a comparative method for estimating the loading of water bodies during peat pruduction

Precipitation-induced runoff and leaching from milled peat mining mires by peat types: a comparative method for estimating the loading of water bodies during peat production. This research project in environmental geology has arisen out of an observed need to be able to predict more accurately the loading of watercourses with detrimental organic substances and nutrients from already existing and planned peat production areas, since the authorities capacity for insisting on such predictions covering the whole duration of peat production in connection with evaluations of environmental impact is at present highly limited. National and international decisions regarding monitoring of the condition of watercourses and their improvement and restoration require more sophisticated evaluation methods in order to be able to forecast watercourse loading and its environmental impacts at the stage of land-use planning and preparations for peat production.The present project thus set out from the premise that it would be possible on the basis of existing mire and peat data properties to construct estimates for the typical loading from production mires over the whole duration of their exploitation. Finland has some 10 million hectares of peatland, accounting for almost a third of its total area. Macroclimatic conditions have varied in the course of the Holocene growth and development of this peatland, and with them the habitats of the peat-forming plants. Temperatures and moisture conditions have played a significant role in determining the dominant species of mire plants growing there at any particular time, the resulting mire types and the accumulation and deposition of plant remains to form the peat. The above climatic, environmental and mire development factors, together with ditching, have contributed, and continue to contribute, to the existence of peat horizons that differ in their physical and chemical properties, leading to differences in material transport between peatlands in a natural state and mires that have been ditched or prepared for forestry and peat production. Watercourse loading from the ditching of mires or their use for peat production can have detrimental effects on river and lake environments and their recreational use, especially where oxygen-consuming organic solids and soluble organic substances and nutrients are concerned. It has not previously been possible, however, to estimate in advance the watercourse loading likely to arise from ditching and peat production on the basis of the characteristics of the peat in a mire, although earlier observations have indicated that watercourse loading from peat production can vary greatly and it has been suggested that differences in peat properties may be of significance in this. Sprinkling is used here in combination with simulations of conditions in a milled peat production area to determine the influence of the physical and chemical properties of milled peats in production mires on surface runoff into the drainage ditches and the concentrations of material in the runoff water. Sprinkling and extraction experiments were carried out on 25 samples of milled Carex (C) and Sphagnum (S) peat of humification grades H 2.5 8.5 with moisture content in the range 23.4 89% on commencement of the first sprinkling, which was followed by a second sprinkling 24 hours later. The water retention capacity of the peat was best, and surface runoff lowest, with Sphagnum and Carex peat samples of humification grades H 2.5 6 in the moisture content class 56 75%. On account of the hydrophobicity of dry peat, runoff increased in a fairly regular manner with drying of the sample from 55% to 24 30%. Runoff from the samples with an original moisture content over 55% increased by 63% in the second round of sprinkling relative to the first, as they had practically reached saturation point on the first occasion, while those with an original moisture content below 55% retained their high runoff in the second round, due to continued hydrophobicity. The well-humified samples (H 6.5 8.5) with a moisture content over 80% showed a low water retention capacity and high runoff in both rounds of sprinkling. Loading of the runoff water with suspended solids, total phosphorus and total nitrogen, and also the chemical oxygen demand (CODMn O2), varied greatly in the sprinkling experiment, depending on the peat type and degree of humification, but concentrations of the same substances in the two sprinklings were closely or moderately closely correlated and these correlations were significant. The concentrations of suspended solids in the runoff water observed in the simulations of a peat production area and the direct surface runoff from it into the drainage ditch system in response to rain (sprinkling intensity 1.27 mm/min) varied c. 60-fold between the degrees of humification in the case of the Carex peats and c. 150-fold for the Sphagnum peats, while chemical oxygen demand varied c. 30-fold and c. 50-fold, respectively, total phosphorus c. 60-fold and c. 66-fold, total nitrogen c. 65-fold and c. 195-fold and ammonium nitrogen c. 90-fold and c. 30-fold. The increases in concentrations in the runoff water were very closely correlated with increases in humification of the peat. The correlations of the concentrations measured in extraction experiments (48 h) with peat type and degree of humification corresponded to those observed in the sprinkler experiments. The resulting figures for the surface runoff from a peat production area into the drainage ditches simulated by means of sprinkling and material concentrations in the runoff water were combined with statistics on the mean extent of daily rainfall (0 67 mm) during the frost-free period of the year (May October) over an observation period of 30 years to yield typical annual loading figures (kg/ha) for suspended solids (SS), chemical oxygen demand of organic matter (CODmn O2), total phosphorus (tot. P) and total nitrogen (tot. N) entering the ditches with respect to milled Carex (C) and Sphagnum (S) peats of humification grades H 2.5 8.5. In order to calculate the loading of drainage ditches from a milled peat production mire with the aid of these annual comparative values (in kg/ha), information is required on the properties of the intended production mire and its peat. Once data are available on the area of the mire, its peat depth, peat types and their degrees of humification, dry matter content, calorific value and corresponding energy content, it is possible to produce mutually comparable estimates for individual mires with respect to the annual loading of the drainage ditch system and the surrounding watercourse for the whole service life of the production area, the duration of this service life, determinations of energy content and the amount of loading per unit of energy generated (kg/MWh). In the 8 mires in the Köyhäjoki basin, Central Ostrobothnia, taken as an example, the loading of suspended solids (SS) in the drainage ditch networks calculated on the basis of the typical values obtained here and existing mire and peat data and expressed per unit of energy generated varied between the mires and horizons in the range 0.9 16.5 kg/MWh. One of the aims of this work was to develop means of making better use of existing mire and peat data and the results of corings and other field investigations. In this respect combination of the typical loading values (kg/ha) obtained here for S, SC, CS and C peats and the various degrees of humification (H 2.5 8.5) with the above mire and peat data by means of a computer program for the acquisition and handling of such data would enable all the information currently available and that deposited in the system in the future to be used for defining watercourse loading estimates for mires and comparing them with the corresponding estimates of energy content. The intention behind this work has been to respond to the challenge facing the energy generation industry to find larger peat production areas that exert less loading on the environment and to that facing the environmental authorities to improve the means available for estimating watercourse loading from peat production and its environmental impacts in advance. The results conform well to the initial hypothesis and to the goals laid down for the research and should enable watercourse loading from existing and planned peat production to be evaluated better in the future and the resulting impacts to be taken into account when planning land use and energy generation. The advance loading information available in this way would be of value in the selection of individual peat production areas, the planning of their exploitation, the introduction of water protection measures and the planning of loading inspections, in order to achieve controlled peat production that pays due attention to environmental considerations.