Epidemiology and pathogenicity of Aleutian mink disease virus

Aleutian mink disease virus (AMDV), species Carnivore amdoparvovirus 1 and family Parvoviridae, causes Aleutian disease (AD), one of the most significant infectious diseases of American mink. It causes high antibody levels, plasmacytosis, and immune complex disease with symptoms ranging from subclinical to fatal. After the first cases were reported in North America in the 1950s, the virus spread to all mink-producing countries and to nature, where it infected several other mustelid and carnivore species. This thesis focused on AMDV epidemiology, evolution, transmission, varying disease severity, and development of polymerase chain reaction (PCR) and sequencing methods to increase understanding on the disease and provide solutions for disease control. Currently, AMDV diagnostics in Finland is based on antibody screening with ELISA and confirmation of positive results with PCR using DNA exctacted from spleen. However, non-specific products caused frequent delays in PCR diagnostics. To update the diagnostics, we validated two used SYBR green-based PCRs (pan-AMDV-PCR and pan-AMDO-PCR) and established a novel probe-based PCR (NS1-probe-PCR). All three PCRs had comparable sensitivities (down to 20 copies/reaction) defined by plasmid dilution series and diagnostic spleen samples. The NS1-probe-PCR was chosen as the primary method due to optimal performance. We also optimized two previously published methods for whole-genome sequencing. Insufficient sequence data for an efficient primer design proved to be an issue with the first protocol relying on PCR amplification of the genome, which is why we modified a metagenomic approach for ssDNA virus sequencing and successfully used it to sequence the entire AMDV genome from five samples. To study the impact of intense farming practices on evolution and transmission of AMDV, we analyzed virus strains from fur farms and free-ranging mustelids from Finland and Poland. Viral sequences indicated that AMDV had appeared in both countries on several separate occasions, and country-specific clustering and frequent transmission between countries were observed. Results from comparison of strains from farmed and free-ranging mink differed between countries. Sequence data from Poland confirmed that farming affects AMDV diversity in the wild within and between study sites. Further sequence analysis identified frequent coinfection and recombination. Two major and several possible recombination breakpoints were identified. The evolution rate of AMDV was estimated to be higher than with most DNA viruses and slightly higher than with other parvoviruses, probably due to farming conditions with many mink being in close proximity. The degree of within-host evolution was higher in free-ranging mink than farmed mink, indicating that they had lived with the virus longer than farmed mink, which are usually culled annually. No notable difference was detected between free-ranging and farmed mink in positive and negative selection patterns in partial NS1 protein. However, a more thorough comparison would require a larger dataset of the complete genomes of both groups than is currently available. American mink and native mustelids from Poland were analyzed for AMDV antibodies with ELISA. The proportion of infected mink was higher in males than females, adults than juveniles, and in areas close to the mink farms, indicating virus flow from farms into the wild. The proportion of ELISA-positive individuals among native mustelids appeared to correlate with the expected contact of the species with the mink. Almost all the ELISA-positive mink were PCR-positive, but only a fraction of the ELISA-positive native mustelids were also PCR-positive, suggesting lower persistence of AMDV in the latter. In addition to being found among native mustelids, AMDV DNA was also detected in stools of farmed blue foxes, indicating that they may be infected and secrete the virus in stool. However, more research is required to exclude the possibility of viruses originating from the contaminated environment or food. In order to study varying disease severity, we compared a farm with mainly asymptomatic mink and a decades-long history of breeding a disease-tolerant herd, a farm with symptomatic mink, and an AMDV-negative farm. Differences in viral loads in spleens and kidneys were observed between asymptomatic and symptomatic mink. In the farm with AMDV-tolerant mink, a notable proportion of ELISA-negative mink were PCR-positive during the 2.5-year follow-up, indicating very low antibody production. Transcriptome analysis revealed differences in gene expression between farms. The results show that mink can live with the virus for years without visible symptoms, have a normal litter size and pelt quality, and bring up breeding of tolerant herd as an option in disease control. In summary, we updated PCR diagnostics to improve speed and reliability that are important in disease control. Additionally, we set up a metagenomic protocol to sequence AMDV without prior sequence information. These studies also brought new information about AMDV epidemiology and host dynamics that can be used to better understand virus history, and transmission between farms and the wild, between countries, and between species. Comparison of mink with a different disease status gave information about AMDV pathogenesis in different stages of infection and gave new insights into disease control.Aleutian tauti -virus (AMDV, Aleutian mink disease virus) aiheuttaa Aleutian tautia, joka on yksi minkin yleisimmistä infektiotaudeista. Se aiheuttaa korkeita vasta-ainetasoja, plasmasytoosia ja immuunikompleksitautia ja oirekuva saattaa vaihdella oireettomasta tappavaan. Tauti havaittiin Pohjois-Amerikassa 1950-luvulla, mutta se on sittemmin levinnyt kaikkiin minkinkasvattajamaihin ja luontoon, jossa se infektoi useita näätä- ja koiraeläinlajeja. Tämän väitöskirjan tavoitteena oli tutkia AMDV:n epidemiologiaa, evoluutiota, leviämistä ja patogeneesiä sekä kehittää PCR- ja sekvensointimenetelmiä avuksi taudin ymmärtämisessä ja kontrolloinnissa. Päivitimme PCR diagnostiikkaa validoimalla kaksi käytössä ollutta SYBG green -pohjaista PCR-menetelmää sekä pystytimme uuden koetinpohjaisen PCR-menetelmän, joka otettiin sittemmin käyttöön diagnostiikassa. Lisäksi optimoimme kahta aiemmin julkaistua menetelmää kokonaisten virusgenomien sekvensointiin ja sekvensoimme kokonaisia virusgenomeja useista näytteistä. Tutkimme turkistarhauksen vaikutusta AMDV:n evoluutioon ja leviämiseen analysoimalla viruskantoja turkistiloilta ja villieläimistä Suomessa ja Puolassa. Virussekvenssien perusteella AMDV on levinnyt molempiin maihin useassa erässä ja sekä maakohtaista ryhmittymistä että runsasta maiden välistä leviämistä oli havaittavissa. Sekvenssidata Puolasta vahvisti, että turkistarhaus vaikuttaa viruksen monimuotoisuuteen ja leviämiseen luonnossa. Havaitsimme myös runsaasti koinfektioita, rekombinaatiota ja poikkeuksellisen nopeaa evoluutiota. AMDV-vasta-aineita verrattiin minkeissä ja muissa näätäeläimissä. Urokset olivat vasta-aine-positiivisia naaraita useammin ja aikuiset nuoria minkkejä useammin. Isompi osa minkeistä oli vasta-ainepositiivisia tilojen lähellä kuin kauempana tiloista viitaten viruksen leviämiseen tiloilta luontoon. Vasta-ainepositiivisten yksilöiden osuus muissa näätäeläimissä korreloi sen kanssa paljonko laji on kontaktissa minkin kanssa ja muut lajit näyttävät pääsevän viruksesta eroon minkkiä useammin. Taudin vakavuuden vaihtelua tutkittiin vertaamalla AMDV-positiivista tolerantteja minkkejä vuosikymmeniä jalostanutta tilaa, jolla ei ollut juurikaan oireista tautia, tilaan, jolla oli oireilevia minkkejä sekä puhtaaseen tilaan. Oireilevilla minkeillä havaittiin merkittävästi enemmän virusta kudoksissa kuin oireettomilla. Tolerantteja minkkejä jalostaneella tilalla iso osa minkeistä oli vasta-aine-negatiivisia ja PCR-positiivisia 2,5 vuoden seurannan aikana viitaten hyvin matalaan vasta-ainetuotantoon. Transkriptominanalyysi paljasti eroja geeniekspressiossa tilojen välillä. Tulokset osoittavat, että minkit voivat elää AMDV:n kanssa vuosia ilman näkyviä oireita. Näissä projekteissa päivitimme AMDV:n leviämisen kontrolloinnissa tärkeää PCR-diagnostiikkaa nopeammaksi ja luotettavammaksi sekä pystytimme metagenomisen menetelmän AMDV:n ja muiden ssDNA-virusten genomien sekvensointiin ilman tarkempaa tietoa virussekvenssistä. Lisäksi hankimme uutta tietoa viruksen epidemiologiasta, mikä mahdollistaa sen historian ja leviämisen paremman ymmärtämisen. Eri taudinkuvien vertailu puolestaan valotti AMDV:n patogeneesiä ja antoi uusia ulottuvuuksia taudin kontrollointiin

Pelasta maailma - kohderyhmiin vaikuttaminen yhteiskunnallisen markkinoinnin keinoin

Siirretty Doriast

Unemployment Trajectories and the Early Risk of Disability Pension among Young People with and without Autism Spectrum Disorder: A Nationwide Study in Sweden

Depression and anxiety are associated with unemployment and disability pension, while autism spectrum disorder (ASD) is less studied. We aimed to first identify unemployment trajectories among young adults with and without ASD, and then to examine their social determinants. Finally, we used the trajectories as determinants for subsequent disability pension. We used a population-based cohort, including 814 people who were 19–35 years old, not on disability pension, and who had their ASD diagnosis between 2001 and 2009. A matched reference population included 22,013 people with no record of mental disorders. Unemployment follow-up was the inclusion year and four years after. Disability pension follow-up started after the unemployment follow-up and continued through 2013. We identified three distinctive trajectories of unemployment during the follow-up: (1) low, then sharply increasing (9%,) (2) low (reference, 67%), and (3) high then slowly decreasing (24%). People with ASD had higher odds of belonging belong to the trajectory groups 1 (OR 2.53, 95% CI 2.02–3.18) and 3 (OR 3.60, 95% CI 3.08–4.19). However, the mean number of unemployment days was relatively low in all groups. A disability pension was a rare event in the cohort, although memberships to groups 1 and 3 were associated with the risk of a future disability pension. More knowledge is needed about factors facilitating participation in paid employment among people with ASD

Menetelmäkuvaus ravinnejalanjäljen laskemiseksi. Laskentaesimerkkinä elintarvikeketju. NUTS-hankkeen loppuraportti

Typpi, fosfori ja kalium, ovat välttämättömiä ravinteita kaikelle turvatulle ruuantuotannolle ja elämiselle. Ihmistoiminnan seurauksena kuitenkin varsinkin typpeä ja fosforia on saatettu ravinnekiertoon liikaa, mikä näkyy tuotantoketjujen ravinnevuotojen vuoksi ravinnepäästöinä ja erilaisina ympäristövaikutuksina. Päästöjen lisäksi resurssien rajallisuus rasittaa ravinnetaloutta: fosforivarannot ovat hupenemassa ja reaktiivisen typen valmistaminen ilmakehästä kuluttaa runsaasti fossiilisia polttoaineita. Jotta edellä oleviin ongelmiin voidaan puuttua yksittäisten tuotanto- ja kulutusketjujen hallinnan kautta, on ravinnevirtoja pystyttävä mittaamaan ja arvottamaan. Tässä työssä esitellään menetelmä ravinnejalanjäljen laskemiseksi. Ravinnejalanjäljellä pyritään mittaamaan erilaisten tuotanto- ja kulutusketjujen ravinteiden käytön tehokkuutta. Sillä tarkastellaan, kuinka paljon ketju ottaa käyttöön ravinnetta, ja kuinka paljon käyttöönotetusta ravinnemäärästä saadaan hyötykäyttöön, joko tarkasteltavaan tuotteeseen tai sivutuotteeseen tai muihin ravinteiden hyödyntävään hyödykkeeseen. Samalla tunnistetaan myös ketjun vuotokohdat: hukatut ravinteet voivat olla ravinnehaihtumia tai -huuhtoutumia, tai ravinteita, jotka päätyvät pitkäaikaiseen varastointiin tai jätteenä muuhun kuin ravinnehyötykäyttöön. Näiden luokitusten avulla lasketaan ravinnejalanjälki tarkasteltua toiminnallista yksikköä kohden: ”Tuotetta varten otetaan käyttöön x kg ravinnetta (typpeä tai fosforia), josta hyödynnettiin tuotteessa y % ja kaikki hyötykäyttö huomioon ottaen z %.”Tässä työssä ravinnejalanjäljen laskeminen demonstroidaan kaurahiutaleelle ja kaurapuurolle tehdyn laskelman kautta. Raportissa on ohjeistettu ravinnejalanjäljen laskenta kussakin elinkaaren vaiheessa alkaen alkutuotannon panostuotannosta ja päättyen jätehuoltoon. Laskelman tulokset osoittavat, että kaurahiutaleketju ottaa käyttöön elinkaarensa aikana 42 kg typpeä ja 7 kg fosforia. Typestä 55 % hyödynnettiin päätuotteessa, siis kaurahiutaleissa, ja kokonaisuudessaan, sivutuotteet huomioon ottaen, typen hyötykäyttöaste oli 71 %. Fosforista 55 % hyödynnettiin päätuotteessa ja kokonaisuudessaan fosforin hyötykäyttöaste oli 99 %. Selkeästi heikoimmat typen ja fosforin hyötykäyttöasteet (alle 50 %) ovat jäteveden käsittelyssä ja elintarvikejätteen käsittelyssä. Laskennan tuloksien perusteella voidaan sanoa, että eniten kehitettävää olisi erityisesti jätehuollossa. Ravinnejalanjälki tarjoaa yksinkertaisessa ja vertailukelpoisessa muodossa tietoa ravinneresurssien kulutuksesta ja käytön tehokkuudesta. Siltä osin se on rinnasteinen vesijalanjäljen kanssa vaikkakaan ei ole ravinteiden suhteellista saatavuutta samalla tapaa huomioon kuin vesijalanjälki. Tehdyn laskelman perusteella voidaan todeta ravinnejalanjäljen osoittavan potentiaalia käyttökelpoisena menetelmänä kestävyyden ympäristöllisen dimension tarkasteluun muiden LCA -pohjaisten menetelmien kuten ilmastonmuutos- ja rehevöitymispotentiaalin arvioinnin rinnalla. Ravinnejalanjäljen, hiilijalanjäljen, rehevöittämispotentiaalin sekä lisäksi biodiversiteetin muutosta kuvaavan potentiaalin ja ekotoksisen jalanjäljen arviointi antaisi yhdistettyinä tuotevirran ja tuotteiden ekologisesta kestävyydestä jo nykyistä paljon paremman kokonaiskuvan. Ravinnejalanjälkeä voidaan soveltaa elintarvikkeiden tuotantotehokkuuden tarkasteluun, mikä on järkevää lannoite- ja ruuantuotannon vastatessa suurimmasta osasta ravinnevirroista, mutta myös se soveltuu myös muihin bioenergian ja biomateriaalien tuotantoketjujen tarkasteluun ja kehittämiseen.ABSTRACT Nitrogen, phosphorus and potassium are necessary nutrients for all secure food production and all life. However, as a result of human activity, excessive amounts of nitrogen and phosphorus in particular have been introduced into the nutrient cycle, which, due to nutrient leakage from production chains, manifests itself in the form of nutrient emissions and different environmental impacts. In addition to emissions, the nutrient economy is burdened by the limited amount of resources available: the phosphorus resources are dwindling, and the manufacture of reactive nitrogen from theatmosphere requires large amounts of fossil fuels. To be able to tackle the problems described above through management of individual production and consumption chains, it must be possible to quantify and evaluate nutrient flows. This study presents a method for calculating the nutrient footprint. The aim with using the nutrient footprint is to measure the efficiency of utilisation of nutrients in various production and consumption chains. It is used for examining how much nutrients each chain takes in, and how much of this intake of nutrients is utilised, either in the product under study or its by-product, or some other commodity making use of nutrients. At the same time, the leakage points along the chain are also identified: the nutrients can be lost through evaporation or leaching, or they may end up in long-term storage, or as waste without nutrient recovery. On the basis of these classifications, the nutrient footprint per functional unit under study is calculated: “For the product, a total of x kg of the nutrient (nitrogen or phosphorus) is taken to use, of which y% was utilised in the product, the total utilisation rate amounting to z% with all practical uses considered.” In this study, the calculation of the nutrient footprint is demonstrated by means of a calculation made for oatmeal and porridge. The report includes instructions for calculating the nutrient footprint in each phase of the life cycle, beginning from input production in primary production and ending with waste management. The results of the calculation show that, during its life cycle, the oatmeal chain takes in 42 kg of nitrogen and 7 kg of phosphorus. 55% of the nitrogen was utilised in the main product, or oatmeal and, with the by-products considered, the total utilisation rate of nitrogen amounted to 71%. 55% of the phosphorus was utilised in the main product, and the total utilisation rate of phosphorus was 99%. Clearly the weakest utilisation rates of nitrogen and phosphorus (less than 50%) are achieved in sewage disposal and food waste treatment. On the basis of the results of this calculation, it could be stated that waste management is the sector with the most room for improvement. The nutrient footprint offers information on the consumption and efficiency of utilisation of nutrient resources in a simple and comparable form. In that sense, it is comparable with the water footprint, even though it does not take account of the relative access to nutrients in the same manner as the water footprint. On the basis of the calculation made, it can be concluded that the nutrient footprint would seem to be a usable method for examining the environmental dimension of sustainability alongside other LCA-based methods, such as the assessment of the climate change potential and eutrophication potential. In addition, we would be able to obtain a much clearer overall image of the ecological sustainability of the product flows and the products if the assessments of the nutrient footprint, the carbon footprint, and the eutrophication potential, as well as the ecotoxicity footprint and the potential describing the biodiversity change were combined. The nutrient footprint can be applied to the study of the production efficiency of food products, which is rational since feed and food production is responsible for the main part of nutrient flows, but it is also suited for the examination and development of other production chains for bioenergy and biomaterials.201

Rapid Absorption of Naloxone from Eye Drops

Naloxone as emergency treatment for opioid overdosing can be administered via several routes. However, the available administration methods are invasive or may be associated with incomplete or slow naloxone absorption. We evaluated pharmacokinetics and local tolerance of naloxone ocular drops in healthy beagle dogs. Naloxone administration as eye drops produced fast absorption with time to maximum plasma concentration (tmax) achieved in 14 to 28 min, high plasma exposure (Cmax 10.3 ng/mL to 12.7 ng/mL), and good bioavailability (41% to 56%). No signs of ocular irritability were observed in the scored ocular tolerability parameters, and the reactions of dogs suggesting immediate ocular discomfort after the dosing were sporadic and short lasting. Slight and transient increase in the intraocular pressure and transient decrease in the tear production were recorded. The results suggest that eye drops may provide a fast and an effective non-invasive route for naloxone administration to reverse opioid overdosing, and clinical studies in the human are warranted

Rapid Absorption of Naloxone from Eye Drops

Naloxone as emergency treatment for opioid overdosing can be administered via several routes. However, the available administration methods are invasive or may be associated with incomplete or slow naloxone absorption. We evaluated pharmacokinetics and local tolerance of naloxone ocular drops in healthy beagle dogs. Naloxone administration as eye drops produced fast absorption with time to maximum plasma concentration (tmax) achieved in 14 to 28 min, high plasma exposure (Cmax 10.3 ng/mL to 12.7 ng/mL), and good bioavailability (41% to 56%). No signs of ocular irritability were observed in the scored ocular tolerability parameters, and the reactions of dogs suggesting immediate ocular discomfort after the dosing were sporadic and short lasting. Slight and transient increase in the intraocular pressure and transient decrease in the tear production were recorded. The results suggest that eye drops may provide a fast and an effective non-invasive route for naloxone administration to reverse opioid overdosing, and clinical studies in the human are warranted

Rapid Absorption of Naloxone from Eye Drops

Naloxone as emergency treatment for opioid overdosing can be administered via several routes. However, the available administration methods are invasive or may be associated with incomplete or slow naloxone absorption. We evaluated pharmacokinetics and local tolerance of naloxone ocular drops in healthy beagle dogs. Naloxone administration as eye drops produced fast absorption with time to maximum plasma concentration (t(max)) achieved in 14 to 28 min, high plasma exposure (C-max 10.3 ng/mL to 12.7 ng/mL), and good bioavailability (41% to 56%). No signs of ocular irritability were observed in the scored ocular tolerability parameters, and the reactions of dogs suggesting immediate ocular discomfort after the dosing were sporadic and short lasting. Slight and transient increase in the intraocular pressure and transient decrease in the tear production were recorded. The results suggest that eye drops may provide a fast and an effective non-invasive route for naloxone administration to reverse opioid overdosing, and clinical studies in the human are warranted.Peer reviewe

Changes in drinking as predictors of changes in sickness absence : a case-crossover study

Background We investigated whether changes in alcohol use predict changes in the risk of sickness absence in a case-crossover design. Methods Finnish public sector employees were surveyed in 2000, 2004 and 2008 on alcohol use and covariates. Heavy drinking was defined as either a weekly intake that exceeded recommendations (12 units for women; 23 for men) or having an extreme drinking session. The responses were linked to national sickness absence registers. We analysed the within-person relative risk of change in the risk of sickness absence in relation to change in drinking. Case period refers to being sickness absent within 1 year of the survey and control period refers to not being sickness absent within 1 year of the survey. Results Periods of heavy drinking were associated with increased odds of self-certified short-term (1-3 days) sickness absence (multivariable-adjusted OR 1.21, 95% CI 1.07 to 1.38 for all participants; 1.62, 95% CI 1.19 to 2.21 for men and 1.15, 95% CI 1.00 to 1.33 for women). A higher risk of short-term sickness absence was also observed after increase in drinking (OR=1.27, 95% CI 1.07 to 1.52) and a lower risk was observed after decrease in drinking (OR=0.83, 95% CI 0.69 to 1.00). Both increase (OR=1.38, 95% CI 1.21 to 1.57) and decrease (OR=1.27, 95% CI 1.19 to 1.43) in drinking were associated with increased risk of long-term (> 9 days) medically certified all-cause sickness absence. Conclusion Increase in drinking was related to increases in short-term and long-term sickness absences. Men and employees with a low socioeconomic position in particular seemed to be at risk.Peer reviewe

Diversity and transmission of Aleutian mink disease virus in feral and farmed American mink and native mustelids

Aleutian mink disease virus (AMDV), which causes Aleutian disease, is widely spread both in farmed mink and wild mustelids. However, only limited data are available on the role of wild animals in AMDV transmission and spread. Our aim was to shed light on AMDV transmission among wild mustelids and estimate the effect of intense farming practices on the virus circulation by studying AMDV prevalence and genetic diversity among wild mustelids in Poland. We compared AMDV seroprevalence and proportion of PCR-positive individuals in American mink, polecats, otters, stone martens, and pine martens and used the phylogenetic analysis of the NS1 region to study transmission. In addition, we used a metagenomic approach to sequence complete AMDV genomes from tissue samples. In eastern Poland, AMDV seroprevalence in wild mustelids varied from 22 per cent in otters to 62 per cent and 64 per cent in stone martens and feral mink, respectively. All studied antibody-positive mink were also PCR positive, whereas only 10, 15, and 18 per cent of antibody-positive polecats, pine martens, and stone martens, respectively, were PCR positive, suggesting lower virus persistence among these animal species as compared to feral mink. In phylogenetic analysis, most sequences from feral mink formed region-specific clusters that have most likely emerged through multiple introductions of AMDV to feral mink population over decades. However, virus spread between regions was also observed. Virus sequences derived from farmed and wild animals formed separate subclusters in the phylogenetic tree, and no signs of recent virus transmission between farmed and wild animals were observed despite the frequent inflow of farmed mink escapees to wild populations. These results provide new information about the role of different mustelid species in AMDV transmission and about virus circulation among the wild mustelids. In addition, we pinpoint gaps of knowledge, where more studies are needed to achieve a comprehensive picture of AMDV transmission.Peer reviewe