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Environmental stress caused by heavy metal contamination of the sediment can threaten ecosystem functioning. Sediment macrofauna are often used to study the effects of environmental stress factors over time, as they are relatively sedentary and thus reflect the ambient conditions in an area. This study investigates whether heavy metal pollution influences the macrofaunal community adjacent to a former steel works factory in Koverhar, in the western Gulf of Finland. Various indices based on macrofaunal community composition and diversity are used in the Baltic Sea to evaluate the environmental status. This thesis evaluates the performance of three of these indices, Shannon-Wiener’s Index (H’), Benthic Quality Index (BQI) and Brackish water Benthic Index (BBI), in detecting the influence of heavy metal pollution on the marine environment. Two macrofaunal sampling methods, GEMAX corer and van Veen grab, are also compared to each other to investigate if there are differences in the structure of the macrofaunal communities that they capture. The study found that while there were indications of environmental stress, such as a lack of sensitive species and an abundance of tolerant species at the more heavily polluted stations, the heavy metal pollution could not be definitively proven to be the cause. H’ and BBI failed to find the differences potentially associated with heavy metal pollution between the stations, while BQI detected some of the differences found by the macrofaunal community analysis. The two sampling methods were found to not be significantly different from each other in terms of macrofaunal communities, but yielded significantly different macrofaunal index values, with the GEMAX results displaying a larger variance between replicates while the van Veen results were more consistent.

Agnico Eagle Ltd. gold mine in Kittilä, Northern Finland has released treated process and dry-ing waters into River Loukinen since December 2020. The effects of incoming mine discharge waters to brown trout behavior and movements in the river were studied for the first time in 2021. The monitoring was repeated in 2023 according to the terms of the environmental permit. The results of the 2023 study were used in this master’s thesis for determining whether the mine discharge waters affect brown trout behavior by causing avoidance of the area, migration hindrance, or changes in movement activity in the vicinity of the discharge pipe. Electrical con-ductivity (EC) was used as an indicator of the water quality changes caused by the discharge water. Twenty brown trout equipped with external transmitters were released at the site of the discharge water pipe in River Loukinen. Fish movements were tracked for 49 days in August - October 2023 with 11 stationary receivers and occasional location of individual fish with a hand-held receiver while boating in the river. Observation of bypasses and fish movement activity were specifically targeted within the discharge area by mounting several receivers in the vicinity of the pipe. Water quality of the river was monitored above and below the discharge pipe, to determine the level of impact of the discharge waters on river water and its possible effects on fish behavior. The river area below the discharge pipe seemed to be favorable habitat for brown trout with moderate flow rate and heterogenous bottom. Several individuals spent long periods below the discharge pipe regardless of the continuous EC impact of the discharge waters. Many of the monitored brown trout moved widely in the river and even to the main river (11 km downstream) yet returning to the vicinity of the discharge pipe. The results indicate no evident disadvantage or altered behavior of brown trout even under the direct influence of mine discharge waters. However, there was some indication of lower movement activity during periods of higher or increasing EC and decreasing river flow, but this is most likely due to the observed strong negative correlation between flow and EC. Increasing river flow is generally known to enhance fish movement. Even if the EC measurements downstream of the discharge pipe were higher than the natural EC of the river, studied brown trout seemed to behave in a species-typical way considering the habitat preferences and seasonal variations, and the mine discharge waters showed no clear impact on fish movements.

Suot ovat tärkeä osa maailmanlaajuista hiilen kiertokulkua, koska ne varastoivat suuria määriä hiiltä eloperäiseen materiaaliin turpeen muodossa, joka muodostuu biomassan hitaasta hajoamisesta kylmän, hapettoman ja matalan pH:n ympäristön vuoksi. Soista vapautuu myös metaania (CH4), joka on voimakas kasvihuonekaasu, jonka lämmityspotentiaali on 28 kertaa voimakkaampi kuin hiilidioksidin (CO2). Turvemaiden netto-C-päästöt riippuvat suotyypistä ja ympäristöolosuhteiden muutoksista, kuten pohjaveden korkeudesta tai turpeen lämpötilasta, ja niistä johtuvasta tasapainosta CH4-päästöjen ja turpeen muodostumisesta johtuvan hiilinielun välillä. Tämän tutkimuksen tavoitteena oli selvittää, miten kasviyhteisöt ja muut säätelevät tekijät, kuten lämpötila, pohjaveden korekus, LAI ja suotyyppi vaikuttavat sekä ilmakehän hiilivirtaan että turpeen CH4- ja CO2-pitoisuuksiin. Lisäksi tehtiin stabiiliin hiili-13 isotoopin mittauksia, jolla saadaan lisätietoa metanogeneesin biogeokemiasta. Mittaukset otettiin rahkasammalvaltaisista mättäistä ja saravaltaisista välipinnoista. Mittauspisteille tehtiin kolme kasvillisuuden manipulointia, joilla selvitettiin kasvillisuuden vaikutuksia hiilidynamiikkaan 1. putkilokasvien ja sammaleiden poisto, 2. pelkkä putkilokasvien poisto, 3. Kaikki kasvillisuus tallella. Tutkimuspaikka sijaitsee Etelä-Suomessa Siikanevan suoalueella. Mittaukset tehtiin vuonna 2018 touko-syyskuussa ombrotrofisessa keidasrämeessä ja oligotrofisessa saranevassa. Mittauskausi oli poikkeuksellisen kuiva ja pohjavedenkorkeus oli keskiarvoa matalammalla. Tästä johtuen monia aikaisemmin havaittuja korrelaatioita ei löytynyt. CH4-virtojen suuruus riippui suotyypistä ja kasvillisuuden manipuloinnista. Keskimääräiset turpeen CH4 ja CO2 pitoisuudet olivat hieman korkeammat mittauspisteissä saranavevalla. Pitoisuudet kasvoivat nopeasti syvyyden myötä, 50 cm:n syvyydessä pitoisuudet olivat useita suuruusluokkia suurempia kuin 7-20 cm:n syvyyksissä korkeimpien, mittausten ollessa yli 500 000 ppm. δ13C-CH4-arvot muuttuivat negatiivisemmiksi tyypillisesti syvyyden myötä, kun hydrogenotrofinen metanogeneesi yleistyi. Kasvillisuuden manipuloinneilla oli vaihtelevia vaikutuksia CH4-vuohon, eikä lehtipinta-alaindeksi osoittanut vahvaa lineaarista korrelaatiota CH4:n kanssa. CH4-virtaus oli myös epäherkkä pohjaveden korkeudelle, mutta kasvien välittämä CH4-kuljetus ei todennäköisesti ollut syynä, koska kasvillisuuden poistokäsitellyt mittauspisteet osoittivat myös samanlaista epäherkkyyttä veden korkeudelle. Putkilokasvien ja sammaleiden poistaminen vähensi yleensä CH4-virtoja. Mättäissä, joissa putkilokasvit oli poistettu, mutta sammaleita ei, oli alhaisimmat CH4-virrat. Yhteenvetona voidaan todeta, että useimmat ympäristömuuttujat eivät osoittaneet vahvaa korrelaatiota CH4:n kanssa. Mikään yksittäinen muuttuja ei selittänyt selvästi eroja CH4-vuossa. Turpeen CH4 ja CO2 pitoisuudet riippuvat voimakkaasti syvyydestä ja suotyypistä. Kasvillisuuden poistaminen tyypillisesti vähensi CH4-virtoja.

My master’s thesis aims to determine the impact of soil treatments and the hemi-parasite Rhinanthus minor (yellow rattle) on the soil properties of newly established perennial wildflower meadow. As urbanization and urban green spaces increase, the need for viable methods for establishing biodiverse meadows on existing lawns grows. I joined a lawn to meadow project based at Lammi Biological Station wherein four soil treatments (untreated, scarified, overturned, and replaced with meadow substrate) were employed in eight meadow blocks. Within those blocks, yellow rattle was sown into half of the plots to determine if it can hamper the growth of competitive grass species. My aim was to explore the meadow blocks’ chemical soil properties (phosphate, nitrate + nitrite, ammonium, total carbon, total nitrogen, and pH). The soil properties were measured using LECO analysis, photometric analysis, and a pH meter. In the newly established meadows at Lammi biological stations, there is evidence that soil properties do change as a result of different soil treatments and the introduction of a hemi-parasitic plant. The initial soil properties show that soil turnover results in increased nitrate + nitrite and decreased total nitrogen compared to untreated meadow soil and lawn controls, respectively. Meadow soils replaced with a substrate exhibited nutrient poor conditions typical of low nutrient preferring meadow plants. The effects of yellow rattle on aboveground community structure are not investigated here, but after the first growing season, its presence increased nitrate + nitrite in the first ten cm of soil. Nitrogen mineralization as a result of grass introduced to soil microbes and nutrient dense yellow rattle leaves may be the cause of these changes to the soil properties. Meadow establishment can take 3 – 5 years, so the use of these methods should continue to be observed. I would expect larger differences to manifest as the experiment continues, namely, decreased soil nutrients as more growing seasons pass.

Agriculture is known to have major environmental impacts through intense use of resources and occupation of land, and it is a significant contributor to climate change and biodiversity loss. Extensive agriculture, as High Nature Value (HNV) farming, benefits biodiversity while providing food and other ecosystem services (ESS). The environmental impacts of HNV ruminant production systems utilizing semi-natural grasslands (SNG) have until now been unknown for Finland and Estonia. SNGs are characterized by high biodiversity and a need for constant management. As livestock production, especially ruminant production, HNV farming on SNGs in the respective countries has potentially considerable environmental impacts whilst simultaneously supporting biodiversity through maintenance of the endangered SNGs. This thesis aims at quantifying and comparing the environmental parameters of global warming potential (GWP), land occupation (LO), and biodiversity, through life cycle assessment (LCA) of HNV livestock production systems in the two neighboring countries of Finland and Estonia and disentangling the drivers for the environmental parameters in both countries. In addition, a GWP for production output (meat) is quantified, and the environmental parameters are considered in relation to the production output of the farms. The results indicate that the HNV ruminant farming systems of Estonia are more extensive and self-sufficient to their production than their Finnish counterparts. Enteric fermentation drives the GWP on the farming system as well as on the product level, while the impact of imported inputs is observed in GWP and LO of Finnish HNV farms. Future research shall focus on determining the environmental impacts of water use and carbon sequestration, as well as the effect of HNV farming on below ground biodiversity. In addition, future research shall include animal welfare aspects to gain a comprehensive understanding of the sustainability of HNV ruminant farming systems.

Recent studies have associated ER stress with various types of hearing loss, such as drug- and noise-induced, age-related, and hereditary hearing loss. However, the research has mostly focused on auditory sensory cell (hair cell) death, and it is not well understood if other molecular mechanisms can drive ER stress-dependent hearing loss. We used Manfflox/flox;Pax2-Cre conditional knockout (cKO) mice under the C57BL/6J (B6) mouse strain to study the effects of genetically-induced chronic ER stress on hearing function. In these mice, the gene coding for mesencephalic astrocyte-derived neurotrophic factor (Manf) has been silenced specifically in the cochlea. Manf is thought to act as an ER homeostasis regulator, and it has shown cytoprotective properties in different disease models both in vitro and in vivo. However, Manf’s mode of action is still poorly understood and even less is known about its function in the inner ear. Previously, cKO mice were found to upregulate ER stress markers in the cochlear hair cells. These mice develop progressive high-frequency hearing loss characterized by high-frequency outer hair cell (OHC) death. However, they have elevated hearing thresholds already at postnatal day 22 (P22) before any OHC death takes place and have elevated hearing thresholds in hearing frequencies where OHCs are retained. Therefore, there has to be another pathological mechanism besides OHC death accounting for the elevations in their hearing thresholds. Hence, we wanted to study the effect of ER stress on the outer hair cell hair bundle structure. The hair bundle is located at the apical pole of the hair cells, and it consists of filamentous actin (F-actin)-filled stereocilia. In mechanotransduction (MET), sound stimuli-induced motions of cochlear fluids cause stereocilia to deflect towards the tallest stereocilia row, allowing for depolarization of hair cells and transformation of mechanical force into electrical signal. Therefore, hair bundle is an essential structure for the hearing function. We used scanning electron microscopy (SEM) and fluorescent microscopy to study OHC hair bundles of cKO mice. We saw disorganization of the bundle structure already at P22. It progressed with age and advanced to strong stereocilia fusion by P56. At this age, all of the high-frequency OHCs of cKO mice displayed stereocilia fusion. We used cochlear whole mounts and immunostainings to study the protein composition of OHC stereocilia of Manf-deficient mice. The base of the stereocilia, termed as the tapering region, contains proteins that link the plasma membrane of stereocilia to their F-actin core, ensuring the cohesion of individual stereocilia. Mutations in these proteins have been associated with stereocilia fusion and hair bundle disorganization. At P56, we saw that stereocilia tapering region proteins radixin (RDX) and myosin 6 (Myo6) were mislocalized from the tapering region towards the apical tips of stereocilia in the high-frequency OHCs of cKO mice. Additionally, we saw that PTPRQ – a tapering region protein that is under normal conditions expressed only in the IHCs of mature cochlea – was upregulated in OHCs of cKO mice, yielding an expression pattern similar to RDX and Myo6. In addition, we used the F-actin probe phalloidin to quantitatively compare F-actin densities in the cuticular plates of cKO and WT mice. Cuticular plate is a structure responsible for attaching stereocilia to hair cell body. It consists of a dense F-actin network and prior studies have associated defects in the cuticular plate composition with hearing loss and stereocilia bundle abnormalities. We found a significant decrease in phalloidin staining intensity in the cuticular plates of high-frequency OHCs of cKO mice, indicating that their cuticular plate F-actin rigidity had been reduced. Together our data shows that Manf deficiency promotes diverse impairments in the OHC hair bundles, consequently inducing hearing loss. To conclude, our study presents novel insights into the complexity of ER stress-induced cochlear pathology. We show that ER stress impairs MET by inducing structural changes in the OHC hair bundle. It appears to be the major reason for hearing loss in the cKO mice, rather than hair cell death. In the future, the impact of Manf deficiency to the inner ear should be further studied. For example, younger and aged cKO mice could be studied to better characterize the progression of Manf deficiency-induced cochlear pathology and hearing loss. Similarly, Manf’s effect on hearing should be studied in other ER stress models to determine its role in the hearing function.

Alzheimer’s disease is the most common form of dementia and one of the highest causes of death worldwide. Recent discovery of lymphatic vessels from the dura mater, the outermost meningeal layer covering the central nervous system, has led to reassessment of the role of lymphatic vessels in neuropathological diseases. The meningeal lymphatic vessels drain macromolecules from the cerebrospinal fluid into the deep cervical lymph nodes and their proper function could be crucial for preventing amyloid-beta aggregation into the brain parenchyma. The function of the meningeal lymphatic vessels is still partly unknown. They have been hypothesized to function as an immune cell hub for the brain and dysfunction of the meningeal lymphatic vessels could lead to immune cell changes in the brain parenchyma. In my thesis, the role of the lymphatic vessels in Alzheimer’s disease was investigated by inducing atrophy of the meningeal lymphatic vessels with VEGF-C depletion in an APdE9 mouse model of Alzheimer’s disease. Single cell sequencing was used to identify the cell types present in the dura mater and in the deep cervical lymph nodes of an Alzheimer’s disease mouse model with and without atrophy of the meningeal lymphatic vessels. The amyloid-beta accumulation was immunohistochemically assessed from the brain and the cognitive decline was studied with behavioral tests. The results showed that atrophy of the meningeal lymphatic vessels did not increase the amount of amyloid-beta in the brain or affect the cognitive decline. The single cell sequencing from the meninges provided a more comprehensive cell atlas than has been published before. It was also found that the atrophy of the meningeal lymphatic vessels was associated with changes in the number of immune cells in the dura mater. The biggest changes were in the number of neutrophils and B-cells, which increased. Further studies are needed to evaluate the role of the meningeal lymphatic vessels in Alzheimer’s disease progression, as the results in this thesis were opposite to the results published before.

Canine hip dysplasia (CHD) is a complex developmental orthopedic disorder particularly common in large size breeds. CHD is characterized by the development of a loose and incongruent hip joint. Affected dogs often suffer from secondary osteoarthritis. Radiographic examination reveals flattening of the femoral head and joint widening. An analogous disorder exists in humans. CHD is inherited quantitatively with suggested involvement of the genes of major effect. Genetic studies utilizing dense SNP arrays have revealed few candidate loci and genes for CHD, including the intronic deletion variant of fibrillin 2 (FBN2) gene in Labrador Retriever breed. FBN2 is a promising candidate gene having a functional role in bone and joint development. Therefore, the objective of this study was to investigate the role of FBN2 in the development of CHD in the Finnish dog population and in additional breeds. The specific aims included establishment of well-phenotyped cohorts of samples from four high-risk breeds, namely Labrador Retriever, Golden Retriever, German Shepherd dog and Bernese Mountain Dog and the assessment of the presence and prevalence of the FBN2 deletion in the Finnish dog population affected by CHD. The study cohorts established here will be later utilized also in genome-wide association studies to identify additional CHD loci. Altogether 220 dogs, out of which 53 were Labrador Retrievers, 41 Golden Retrievers, 79 German Shepherd dogs and 47 Bernese Mountain Dogs, were included in the case-control study. For the German Shepherd dogs, the hip status of each dog was further confirmed of by screening the radiographs available from the Finnish Kennel Club archive. All dogs were genotyped by fragment analysis for the FBN2 deletion. The results revealed the presence of the deletion in all four breeds with highest prevalence in the Retriever breeds, in which the deletion haplotype was more common than the wild type. In our study, all CHD-affected dogs in the Labrador Retriever breed had at least one copy of the deletion allele. However, since the deletion allele was common also in the unaffected Labrador Retrievers, no statistically significant allelic association with the deletion and CHD was detected in statistical analysis. In the three other breeds, no association in statistical analysis was found either. Thus, the previously reported positive allelic association between the intronic deletion in the FBN2 gene and CHD was not replicated. Larger genome-wide studies are warranted to identify the major effect CHD loci.

Background: Despite the well-established link between diabetes and dementia risk, the impact of prediabetes and diabetes on the prodromal dementia phase remains controversial. In this study, we investigated whether prediabetes and diabetes increase the risk of cognitive impairment–no dementia (CIND) and accelerate its progression to dementia, as well as the possible underlying mechanisms. Methods: In the Swedish National Study on Aging and Care-Kungsholmen (SNAC-K), one cohort of cognitively-intact individuals (n=1,837) and one cohort of individuals with CIND (n=671) aged ≥60 years were followed for up to 15 years. At baseline and each follow-up (every 3 or 6 years), a neuropsychological test battery was administered, and the domains of episodic memory, processing speed, executive function, visuospatial abilities, and language were derived. CIND was defined as having no dementia and cognitive performance ≤1.5 SDs below age group-specific means in at least one cognitive domain. Dementia was diagnosed according to DSM-IV criteria. Diabetes (controlled and poorly-controlled) was diagnosed by physicians through medical assessment, clinical records, and glycated hemoglobin (HbA1c) ≥6.5%. Prediabetes was identified as HbA1c 5.7-6.4% in diabetes-free participants. Clinicians diagnosed heart disease and collected blood samples used to measure C-reactive protein (CRP). Data were analyzed with Cox regression models adjusted for possible confounders. Results: At baseline, in the cognitively-intact cohort, 133 (7%) participants had diabetes and 615 (34%) had prediabetes. During follow-up (mean 9.2 ± 3.0 years [range=2.2-15.5 years]), 544 (30%) individuals in the cognitively-intact cohort developed CIND. Poorly-controlled diabetes (HbA1c ≥7.5%) was associated with 2-times higher risk of CIND (HR 2.0, 95% CI:1.11-3.48) than diabetes-free participants. In the CIND cohort, 84 (13%) had diabetes and 238 (36%) prediabetes. During follow-up (mean 7.7 ± 4.0 years [range=0.2-15.2 years]), 132 (20%) individuals progressed to dementia. Poorly-controlled diabetes was associated with 3-times higher risk of dementia progression (HR 3.3, 95% CI: 1.29-8.33). Furthermore, comorbid heart disease and diabetes was associated with 2.5-times higher risk of progression to dementia (HR 2.5, 95% CI: 1.17-5.47), particularly if the diabetes was poorly-controlled (HR 5.8, 95% CI: 1.72-19.3). Similarly, having elevated CRP levels and diabetes was associated with increased risk of progression to dementia (HR 4.1, 95% CI: 1.15-14.2), especially in participants with poorly-controlled diabetes (HR 13.6, 95% CI: 1.89-98). No associations between prediabetes and CIND were detected in either cohort. Conclusions: Diabetes, especially if poorly-controlled, increases the risk of cognitive impairment and accelerates its progression to dementia. The diabetes-associated progression from CIND to dementia is further exacerbated by the presence of heart disease and elevated levels of systemic inflammation.

Non-native species can have complex effects on the abundance of native species potentially altering the functioning of ecosystems negatively. Invasive species can outcompete local species competing for resources, ultimately causing the extinction of local species. Inter- and intraspecific competition can be especially vigorous for limited resources. Invasive species have been thought to be a leading cause in native species extinction, and their effects on native species can be especially pronounced during reproductive crucial life-history stages, such as nest-building. Based on previous information about invasive species and their effects on ecosystems, and previous studies conducted related to invasive species, I conducted an experiment at the Tvärminne zoological station in Hanko, southern Finland during May and June of 2021. I conducted a laboratory experiment in which the test species used were the invasive fish species round goby, that has increased its range across the Baltic Sea rapidly, and the native fish species sand goby. The purpose was to see, if there was any effect the invasive species has on the nesting success and motivation of the native species. Methods included five different treatments in aquariums. The results did not differ statistically between different treatments, length was close to statistical significance. However, these results do not demonstrate, that the round goby has no effect on the nest building motivation of sand gobies. Some factors of the experimental setup might have been faulty, and future studies with a larger sample are needed to examine the effects of competition on native species’ abundance.