Extra-Cytoplasmic Function (ECF) σ factors are a subset of σ factors many organisms use to transcribe specific genes in response to environmental cues. In the absence of an inducing signal, ECF σ factors are inhibited by an anti-σ factor that prevents the ECF σ factor from interacting with RNA polymerase. The ECF σ factor σV from Bacillus subtilis is activated in response to lysozyme stress. Lysozyme damages bacterial peptidoglycan by cleaving at the 1,4-ß-linkage between N-acetylmuramic acid and N-acetylglucosamine. In the absence of lysozyme, the activity of σV is inhibited by the anti-σ factor RsiV, a single pass transmembrane protein. The two main components of this project have been to elucidate the mechanism of σV activation and examine how this system senses lysozyme stress. In chapter 2 we show that the activation of σV is specific to lysozyme, and the anti-σ factor RsiV is degraded by a step wise proteolytic cascade known as Regulated Intramembrane Proteolysis (RIP). In the presence of lysozyme, the extracellular domain of RsiV is removed by cleavage at site-1. Upon removal of the extracellular domain, the site-2 protease, RasP, cleaves RsiV within the membrane. The remainder of RsiV is degraded by cytosolic proteases allowing σV to interact with RNA polymerase. In response to lysozyme stress σV activates an O-actyltransferase, OatA, which modifies the peptidoglycan to prevent further lysozyme damage. Our studies in chapter 3 identifed the protease(s) responsible for site-1 cleavage of RsiV and revealed RsiV directly interacts with lysozyme. We determined the cleavage site of the site-1 protease using N-terminal sequencing, and demonstrate that disruption of site-1 cleavage blocks σV activation. Site-1 cleavage occurs at what appears to be a signal peptide cleavage site. We demonstrate that four out of the five signal peptidases from B. subtilis are able to cleave RsiV at site-1 in vitro only in the presence of lysozyme. Additionally, we show that the extracellular domain of RsiV directly binds the inducing substrate lysozyme. In chapter 4 we focus on determining if the interaction between RsiV and lysozyme is necessary for σV activation. Based on the co-crystal structure of RsiV and lysozyme we mutated sveral residues predicted to be involved in binding. One combination of RsiV mutations (S169W, P259A, Y261A) was unable to activate σV and subsequently was unable to bind lysozyme. We propose a RIP dependent mechanism of σV activation that is contingent upon the anti-σ factor (RsiV) directly binding the inducing signal (lysozyme) to present the site-1 cleavage site to signal peptidase. The co-crystal structure of RsiV and lysozyme also revealed that RsiV interacts with the active site of lysozyme. We demonstrate that purified RsiV inhibits lysozyme activity suggesting RsiV provides an additional lysozyme response mechanism. Thus, the anti-σ factor RsiV senses the presence of lysozyme, activates σV, and protects against further lysozyme damage.

The objective of this study was to examine the effects of presynchronization and recombinant bovine somatotropin (rbST) on conception rates following a timed insemination (TAI) protocol in lactating dairy cows. A further objective included the evaluation of the efficacy of the Early Conception Factor (ECF) test kit. Recombinant bST may offer some benefit when used in conjunction with estrus synchronization and TAI. Presynchronization treatment consisted of two injections of PGF2α given 14 d apart, with the second dose administered 14 d prior to the initiation of a TAI protocol. A total of 216 lactating Holstein cows were presynchronized with PGF2α and then received GnRH (100μg) at 67 ± 7 d post partum (PP), administration of PGF2α (25 mg) 7 d later, another GnRH (100μg) administration 2 d after PGF2α, and were inseminated 8-18h later (OvSynch). First service conception rate (CR) was determined by rectal palpation at 42 ± 7 d after artificial insemination (AI). Treated cows (n=113) received rbST 67 ± 7 d PP whereas control cows (n = 113) were presynchronized without rbST. The cycling status of all cows was determined by paired milk P4 levels at 53 and 67 ± 7 d PP. No differences (P > 0.10) in conception rate were observed between control and rbST treated cows (44.7 and 40.7% respectively), nor was there any interaction of cyclicity and rbST. Milk samples were collected 7 d following AI for use in ECF test kit evaluation. Samples were stored at -20ºC (n=216) and at 5ºC (n=113) until assayed. Test results for frozen and refrigerated samples were compared to conception rates determined by rectal palpation at 42 ± 7 d after AI. The rate of false positive and negative results for frozen milk samples were 36.1 and 14.8% respectively, and 40.7 and 7.1% for refrigerated milk samples. Treatment with rbST at the time of the first GnRH injection of an OvSynch protocol did not significantly alter first service conception rates. Additionally, an acceptable 92.9% accuracy of the ECF test for the detection of open cows 7 d after AI using milk samples stored at 5ºC was obtained. / Master of Science

A proteobactéria Pseudomonas aeruginosa é um patógeno oportunista em humanos, sendo associado a queimaduras e infecções pulmonares crônicas em pacientes com fibrose cística. Essas infecções são difíceis de erradicar devido à resistência intrínseca de P. aeruginosa a antibióticos e à formação de biofilmes. Essa bactéria é altamente capaz de adaptar ao ambiente, tem um metabolismo versátil e pode direcionar a expressão de genes por vários fatores sigma alternativos. Estes são subunidades para transcrição de conjuntos específicos de genes em bactérias e interagem com o cerne da RNA polimerase, levando ao reconhecimento do promotor e início da transcrição. Os fatores sigma alternativos permitem que bactérias redirecionem a sua expressão genética. Um grupo de fatores sigma alternativos é o grupo dos fatores sigma de função extracitoplasmática (ECF) que são envolvidos principalmente em funções do envelope celular. Esse trabalho teve como objetivo caracterizar dois fatores sigma ECF de função desconhecida, PA14_21550 e PA14_46810. A linhagem mutante Δ21550 foi analisada quanto a sua sobrevivência a diferentes estresses, observando-se que é mais resistente ao choque de 45°C que a linhagem selvagem. Esse fator sigma não é essencial para crescimento da bactéria em meio LB e meio mínimo M63 acrescido de glicose ou succinato. Além disso, observou-se que a superexpressão desse fator sigma aumenta a expressão da proteína hipotética PA14_30100, usando-se uma abordagem proteômica. O mutante de transposon para o fator sigma PA14_46810 apresenta melhor crescimento que a bactéria selvagem em meio M63 acrescido de glicose. Essa linhagem mostrou mesmo fenótipo para biofilme e formação de exopolissacarídeo que a bactéria selvagem. Ademais, foi realizada análise de transcritoma por RNA-Seq com a superexpressão do fator sigma PA14_46810 na linhagem selvagem. Na linhagem de superexpressão Observou-se que ocorre indução de genes envolvidos com a desnitrificação, transporte de moléculas e metabolismo de uma maneira geral, em relação à linhagem controle. Por outro lado, o excesso de PA14_46810 reprime principalmente genes envolvidos com a tradução de proteínas e síntese de espermidina. Este trabalho, portanto, trouxe novas informações sobre as funções de diferentes fatores sigma ECF de P. aeruginosa, contribuindo assim para um maior entendimento da fisiologia desta bactéria e sua adaptação a diferentes condições. / The proteobacterium Pseudomonas aeruginosa is an opportunistic pathogen in humans, and it is associated to chronic pulmonary infections in patients with cystic fibrosis and burn wounds. These infections are difficult to eradicate due to P. aeruginosa intrinsic resistance to antibiotics and formation of biofilms, which allow the bacteria to adhere to biotic and abiotic surfaces. This bacterium is highly adaptaptable to the environment has a versatile metabolism and can direct the expression of genes by several alternative sigma factors. The sigma factors bind to the RNA polymerase core, providing recognition to promoter and transcription initiation. Therefore, the alternative sigma factors can redirect bacterial genetic expression by recognizing specific promoters. One subfamily of alternative sigma factors is the extracytoplasmic function (ECF) sigma factors, involved mostly in cell envelope functions. The aim of this work was characterize two ECF sigma factors with unknown function in P. aeruginosa, PA14_21550 and PA14_46810. The strain Δ21550 was analyzed for its survival in different stress conditions and it is more resistant in heat shock conditions at 45°C than the wild type strain. It was also observed that PA14_21550 sigma factor is not essential for bacterial growth in LB and M63 minimal medium added with glucose or succinate as the carbon source. Furthermore, overexpression of this sigma factor increases the expression of hypothetical protein PA14_30100, as verified by a proteomic approach. A strain insertionally inactivated in the PA14_46810 gene has better growth than the wild type strain in M63 added with glucose and the same phenotype regarding to biofilm formation and exopolysaccharide production as the wild type strain. Moreover, transcriptome analysis was carried out by RNA-Seq with overexpression of the PA14_46810 sigma factor in the wild type strain. Induction of genes involved in denitrification, transport of molecules and energetic metabolism in relation to the control strain was observed. On the other hand, excess of PA14_46810 represses genes involved in protein translation and spermidine synthesis. This work, therefore, brought new information about the functions of two ECF sigma of P. aeruginosa, thus contributing to a greater understanding of the physiology of this bacterium and its adaptation to different conditions.

A proteobactéria Pseudomonas aeruginosa é um patógeno oportunista em humanos, sendo associado a queimaduras e infecções pulmonares crônicas em pacientes com fibrose cística. Essas infecções são difíceis de erradicar devido à resistência intrínseca de P. aeruginosa a antibióticos e à formação de biofilmes. Essa bactéria é altamente capaz de adaptar ao ambiente, tem um metabolismo versátil e pode direcionar a expressão de genes por vários fatores sigma alternativos. Estes são subunidades para transcrição de conjuntos específicos de genes em bactérias e interagem com o cerne da RNA polimerase, levando ao reconhecimento do promotor e início da transcrição. Os fatores sigma alternativos permitem que bactérias redirecionem a sua expressão genética. Um grupo de fatores sigma alternativos é o grupo dos fatores sigma de função extracitoplasmática (ECF) que são envolvidos principalmente em funções do envelope celular. Esse trabalho teve como objetivo caracterizar dois fatores sigma ECF de função desconhecida, PA14_21550 e PA14_46810. A linhagem mutante Δ21550 foi analisada quanto a sua sobrevivência a diferentes estresses, observando-se que é mais resistente ao choque de 45°C que a linhagem selvagem. Esse fator sigma não é essencial para crescimento da bactéria em meio LB e meio mínimo M63 acrescido de glicose ou succinato. Além disso, observou-se que a superexpressão desse fator sigma aumenta a expressão da proteína hipotética PA14_30100, usando-se uma abordagem proteômica. O mutante de transposon para o fator sigma PA14_46810 apresenta melhor crescimento que a bactéria selvagem em meio M63 acrescido de glicose. Essa linhagem mostrou mesmo fenótipo para biofilme e formação de exopolissacarídeo que a bactéria selvagem. Ademais, foi realizada análise de transcritoma por RNA-Seq com a superexpressão do fator sigma PA14_46810 na linhagem selvagem. Na linhagem de superexpressão Observou-se que ocorre indução de genes envolvidos com a desnitrificação, transporte de moléculas e metabolismo de uma maneira geral, em relação à linhagem controle. Por outro lado, o excesso de PA14_46810 reprime principalmente genes envolvidos com a tradução de proteínas e síntese de espermidina. Este trabalho, portanto, trouxe novas informações sobre as funções de diferentes fatores sigma ECF de P. aeruginosa, contribuindo assim para um maior entendimento da fisiologia desta bactéria e sua adaptação a diferentes condições. / The proteobacterium Pseudomonas aeruginosa is an opportunistic pathogen in humans, and it is associated to chronic pulmonary infections in patients with cystic fibrosis and burn wounds. These infections are difficult to eradicate due to P. aeruginosa intrinsic resistance to antibiotics and formation of biofilms, which allow the bacteria to adhere to biotic and abiotic surfaces. This bacterium is highly adaptaptable to the environment has a versatile metabolism and can direct the expression of genes by several alternative sigma factors. The sigma factors bind to the RNA polymerase core, providing recognition to promoter and transcription initiation. Therefore, the alternative sigma factors can redirect bacterial genetic expression by recognizing specific promoters. One subfamily of alternative sigma factors is the extracytoplasmic function (ECF) sigma factors, involved mostly in cell envelope functions. The aim of this work was characterize two ECF sigma factors with unknown function in P. aeruginosa, PA14_21550 and PA14_46810. The strain Δ21550 was analyzed for its survival in different stress conditions and it is more resistant in heat shock conditions at 45°C than the wild type strain. It was also observed that PA14_21550 sigma factor is not essential for bacterial growth in LB and M63 minimal medium added with glucose or succinate as the carbon source. Furthermore, overexpression of this sigma factor increases the expression of hypothetical protein PA14_30100, as verified by a proteomic approach. A strain insertionally inactivated in the PA14_46810 gene has better growth than the wild type strain in M63 added with glucose and the same phenotype regarding to biofilm formation and exopolysaccharide production as the wild type strain. Moreover, transcriptome analysis was carried out by RNA-Seq with overexpression of the PA14_46810 sigma factor in the wild type strain. Induction of genes involved in denitrification, transport of molecules and energetic metabolism in relation to the control strain was observed. On the other hand, excess of PA14_46810 represses genes involved in protein translation and spermidine synthesis. This work, therefore, brought new information about the functions of two ECF sigma of P. aeruginosa, thus contributing to a greater understanding of the physiology of this bacterium and its adaptation to different conditions.

This work aimed to determine how effective an SVP-LITE® chlorine dioxide plant at Vallviks Bruk was running. This was done by determining the chemical consumption per unit of chlorine dioxide produced. The calculations relied upon online instrumentation that was manually validated. The most important parameters were the conversion ratio of sodium chlorate into chlorine dioxide and the specific consumption of sodium chlorate, methanol, and sulphuric acid. The salt cake filter and dissolver were also examined to determine their impact. The total conversion ratio of sodium chlorate during a whole ECF (Elementally Chlorine Free) campaign were 93.12 %, somewhat lower than the target efficiency of 95.64 %. The specific chemical consumption was 1.695 kg sodium chlorate, 0.183 kg methanol, and 1.007 kg sulphuric acid per kg produced chlorine dioxide. The salt cake filter worked as intended to remove excess sulphuric acid, with the precipitate having the same sulphate and acidic content as the wanted form of sodium sesquisulphate Na3H(SO4)2. However, some sodium chlorate can get through, and the mean concentration in the dissolver was 1.3 g/dm3, equal to approximately 4 tons lost during a campaign. The filter may be more efficient in redissolving acidic species than chlorate salts. The reactor could theoretically be run more efficiently, but its production parameters is close to what is expected of such a plant. / Målet med det här arbetet var att undersöka effektiviteten av en SVP-LITE® klordioxidanläggning vid Vallviks Bruk. SVP-LITE® är en klordioxidproduktionslösning från EKA®. Det gjordes genom att undersöka konsumtionen av reagens per enhet producerad klordioxid. Beräkningarna gjordes från online-instrument som validerades manuellt. De viktigaste parametrarna var konverteringsgraden av natriumklorat till klordioxid samt den specifika förbrukningen av natriumklorat, metanol och svavelsyra. Sulfatlösaren undersöktes för att se hur saltkakefiltret fungerade och dess påverkan. Den totala konverteringsgraden av klorat till klordioxid under en hel ECF (Elementally Chlorine Free) kampanj var 93,12 %, något lägre än måleffektiviteten på 95,64 %. ECF är ett paraplybegrepp för att benämna blekning som inte använder klorgas men som kan använda andra kemikalier som innehåller klor, oftast klordioxid. Den specifika konsumtionen av reagens var 1,695 kg natriumklorat, 0,183 kg metanol och 1,007 kg svavelsyra per kg producerad klordioxid. Saltkakefiltret fungerade som tänk att ta bort överflödig svavelsyra och fällningen hade samma syrlighet som natriumseskvisulfat (Na3H(SO4)2. Natriumklorat kommer dock ut genom filtret och har i upplösaren medelkoncentrationen 1,3 g/dm3 vilket motsvarar en förlust på ca 4 ton under en kampanj. Det är möjligt att filtret är bättre att återupplösa syrliga specier än kloratsalter. Även om reaktorn teoretiskt kan köras mer effektivt är de nuvarande produktionsparametrarna nära de som kan förväntas av en sådan anläggning.

Porphyromonas gingivalis, a gram-negative anaerobic, pathogenic bacterium is a major etiological agent in the initiation and progression of periodontal disease. Due to the ever-changing environment of the oral cavity, inhabitants like Porphyromonas gingivalis must possess the ability to adapt to changes in environmental conditions like pH, temperature, oxygen tension, and metal concentration. P. gingivalis should therefore have an efficient regulatory system in order to adapt and survive in the oral cavity. This response adaptation occurs at the transcriptional level, which involves alternative sigma factors. Extracytoplasmic function sigma (ECF-s) factors are the largest group of alternative sigma factors that play a role in the bacterial response to environmental stress conditions. Here we analyze the s-70 factor gene, PG0214, an extracytoplasmic function sigma factor encoded in the P. gingivalis genome, and examine its role in the bacterial response to environmental stress and virulence. Our findings indicate that the PG0214 gene is important in regulating major functional gene groups and pathways in the P. gingivalis genome. Strains deficient in the PG0214 gene were analyzed and shown to have decreased protease activity, as well as reduced survivability and invasion rates in eukaryotic host cells when compared against wild-type W83 and ATCC 33277 strains. Collectively our studies demonstrate that the PG0214 gene is a positive regulator of gene expression for the survival and virulence of P. gingivalis in the presence of oxidative- and iron-stress, although further study is needed to fully characterize the gene and determine its specific function.

An extensive compulsory cattle dipping programme was introduced in Venda from 1915 to counter rinderpest and East Coast Fever (ECF). This study approached the sustainability of cattle dipping from environmental, economic, social and governance perspectives against the backdrop of the country’s history and political ecology, focusing on the effects of dipping strategies and operations in Vhembe district of Limpopo Province (the former Venda) in response to ECF. Dipping infrastructre continues to be used in Venda after the eradication of ECF in 1954 and even to the present, albeit below capacity. Arsenic residues occur in soils around all sampled dip sites, especially within 20 m from dip tanks and where red clays and organic-rich loamy soils prevail. Ecologically, dipping practice has therefore not been sustainable, while economically, farmers perceive dipping to enhance livestock health, and they gain benefit from continued use of cattle in agriculture and transport. Being close to watercourses and villages, most dip tank sites pose community safety and health risks, with even fatalities occurring at untended and abandoned tank facilities. Yet the social sustainability benefits of dipping practice, such as the creation of forums where cattle owners and veterinarians interact, the resultant formation of cattle owners’ associations, and the occurrence of recreational opportunities on dip days are also evident. Government has been the main role player in providing dipping services, resulting in limited involvement of and cooperation between other role players; therefore the contribution of the Limpopo Draft Policy on Cattle Dipping (2011) to efficient dipping governance was evaluated. A Sustainable Community Cattle Dipping Model was consequently developed to address the shortcomings in governance, as well as ecological, economic and social issues of the sustainability of dipping practice. This thesis contributes to an understanding that cattle dipping in communal areas are spatial entities that reflect the dynamics of structure-agency. It underscores environmental injustices like arsenic contamination occurring around dip tanks, and the effects of inequitable distribution of dip sites on human health and safety. The economic benefits and limitations of existing dipping practice are also highlighted. It culminates in the development of a Sustainable Community Cattle Dipping Model to enhance the sustainability of dipping practice. / Thesis (PhD)--University of Pretoria, 2014. / Geography, Geoinformatics and Meteorology / PhD / Unrestricted

No description available.

SCA Massa, Östrands massafabrik har två produktionslinjer, fiberlinjen som tillverkar NBSK (blekt barrsulfatmassa) och CTMP-linjen som tillverkar CTMP (kemisk termomekanisk massa). Massaindustrin har trotts övergången från klorblekning till klordioxidblekning fortfarande en påtaglig miljöpåverkan, då tillverkningsprocessen medför både restavfall och förorenade utsläpp. Barrsulfatmassan bleks antingen med ECF (Elemental Chlorine Free) eller TCF (Total Chlorine Free) metoden. Under tillverkningsprocessen av ECF-massa så tillsätts klordioxid som blekkemikalie, vilket leder till att avloppsvattnet som bildas innehåller stora mängder kloratjoner och måste renas innan det släpps ut till recipienten. Under 2014 ansökte SCA om nytt verksamhetstillstånd i Mark- och Miljödomstolen, för att kunna bygga ut Östrands massafabrik och producera upp till 1,1 miljon ton pappersmassa per år. Under 2015 beslutade domstolen om ett nytt tillstånd och för kloratutsläpp sattes ett provisoriskt villkor på 0,5 kg klorat/ ton ECF-massa, som kombinerades med två utredningsnivåer. Utredningsnivå 1 på 0,15 kg klorat/ton ECF-massa och utredningsnivå 2 på 0,05 kg klorat/ton ECF-massa.  Syftet med studien var att undersöka hur det biologiska reningssteget påverkas vid växlingar mellan TCF- och ECF-blekning, och vilka möjligheter det finns att reducera risken för ökade kloratutsläpp. Syftet var också att undersöka om SCA uppfyller det provisoriska villkoret med avseende på utgående mängd klorat per tillverkad ton ECF-massa, och vart SCA ligger i förhållande till utredningsnivå 1 respektive 2. Dygnsmedelvärdesprover har sammanställts och utvärderas för totalt fem månader, från december 2020 till april 2021. Dessa har använts för att undersöka den ingående och utgående mängden klorat från reningen och faktorer som kan påverka reningen. Parametrar som undersökts är pH, temperatur, flöde, syrehalt i kloratsteg A och B och pumpgropen. Stickprover har tagits ur blekeriavloppet, bärarsteg 1A och 1B i samband med växlingar från TCF- till ECF-massa för att undersöka hur kloratsteg A och B påverkas.  Resultatet av undersökningen visade att reningsanläggning har en reduktionsgrad av klorat på ca 89 % i de kloratreducerande stegen, och att reduktionsgraden från bärarsteg 1A till 1B är på ca 31 %. Den största reduktionen av klorat sker i blekerireningen men det sker även en viss reduktion i selektorsteget och de fyra seriekopplade aktivslambassängerna. SCA Östrand ligger som medeltal på ett utsläpp av 0,11 kg klorat/ton ECF-massa. Detta innebär att det provisoriska villkoret på 0,5 (kg klorat/ton ECF-massa) och utredningsnivå 1 på 0,15 (kg klorat/ton ECF-massa) uppnås under den undersökta perioden. SCA Östrand uppfyller inte vid något tillfälle under den undersökta perioden utredningsnivå 2 på 0,05 (kg klorat/ ton ECF- massa). Syrehalten i kloratsteg A och B har en avgörande roll för reduktionen av klorat. Under en tidpunkt uppstod ett handhavande fel på syrgasgivaren och syrehalten reglerades inte till en lägre nivå trotts ECF-massa tillverkning, detta bidrog till högre kloratutsläpp på grund av den minskade reduktionen. I bärarsteg 1A visade det sig att 48 % av mängden klorat i härstammade från pumpgropen. För att minimera risken för ökade kloratutsläpp till recipienten bör inkommande processberört vatten från samlingsavloppet ses över, vilket innebär att kontrollera och minska mängderna klorat till pumpgropen kraftigt. SCA Östrand bör se över breddningen av filtertank 8 som bidrar till ökade mängder klorat till pumpgropen, i samband med klordioxidblekning i blekeriet. För att SCA Östrand ska uppnå utredningskrav 2 måste processinterna åtgärder ses över för att minimera risken att processberört vatten med stora mängder klorat inte passerar kloratsteg A och B i blekerireningen. / SCA Massa, Östrand pulp mill is covered by two production lines, the fiber line producing NBSK and the CTMP line producing CTMP. The pulp industry, despite the transition from chlorine bleaching to chlorine dioxide bleaching, still has a significant environmental impact, as the manufacturing process generates both residual waste and polluted emissions. The sulphate mass is bleached using either the ECF (Elemental Chlorine Free) or TCF (Total Chlorine Free) method. During the production process of the sulphate pulp ECF, chlorine dioxide is added as a bleaching chemical, which results in the wastewater produced containing large amounts of chlorate ions and must be treated before being discharged to the recipient. In 2014, SCA applied to the Land and Environmental Court for a new operating permit to expand the Östrand pulp mill and produce up to 1.1 million tons of pulp. In 2015, the court decided on a new permit and a provisional condition of 0.5 kg chlorate/ton ECF-mass was set for chlorate emissions, which was combined with two investigation levels: investigation level 1 of 0.15 kg chlorate/ton ECF-mass and investigation level 2 of 0.05 kg chlorate/ton ECF-mass.  The aim of the study was to investigate how the biological purification step is affected by changes between TCF and ECF bleaching, and what opportunities there are to reduce the risk of increased chlorate emissions. It also aimed to investigate whether SCA meets the provisional requirement with respect to the output of chlorate per tons of ECF mass produced, and where SCA stands in relation to investigation levels 1 and 2. Daily average samples were compiled and evaluated for a total of five months, from December 2020 to April 2021. These were used to investigate the input and output of chlorate from the purification process and factors that may affect the purification. Parameters investigated include pH, temperature, flow, oxygen supply in chlorate stages A and B and the pump pit. Samples were taken from the bleach drainer, carrier stages 1A and 1B, during changes from TCF to ECF mass to investigate the influence of chlorate stages A and B.  The results of the study showed that the facility of purification has a chlorate reduction degree of 89 % in the chlorate-reducing stages, and that the degree of reduction from carrier step 1A to 1B is about 31 %. The largest reduction of chlorate occurs in the bleaching purification, but there is also some reduction in the selector stage and the four series-connected activated sludge units. SCA Östrand has an average emission of 0.11 kg chlorate/ton ECF-mass. This means that the provisional condition of 0.5 (kg chlorate/ton ECF-mass) and investigation level 1 of 0.15 (kg chlorate/ton ECF-mass) are achieved during the periods under investigation. SCA Östrand does not meet investigation level 2 of 0,05 (kg chlorate/ton ECF-mass) at any time during the period under investigation. The oxygen supply in chlorate steps A and B plays a decisive role in the reduction of chlorate. During one point in time a handling error occurred on the oxygen sensor and the oxygen content was not regulated to a lower level despite ECF- mass production, this error contributed to higher chlorate emissions due to the reduced reduction. To minimize the risk of increased chlorate emissions to the recipient, incoming process treated water from collection drains to carrier stage 1A of the bleach plant should be reviewed. It was found that 48 % of the amount of chlorate in carrier stage 1A originated from the pump pit. In order to minimize the risk of increased chlorate discharge, the amounts of chlorate entering the pump pit must be controlled and greatly minimized. SCA Östrand should review the widening of filter tank 8, which contributes to increased amounts of chlorate to the pump pit, in connection with chlorine dioxide bleaching in the bleaching plant. In order for SCA Östrand to meet investigation requirement 2, in-process measures must be reviewed to minimize the risk that process contaminated water with large amounts of chlorate does not pass through chlorate stages A and B in the facility of purification.

Pseudomonas aeruginosa é uma proteobactéria do grupo gama muito versátil, capaz de colonizar ambientes variados e infectar hospedeiros filogeneticamente distintos, incluindo humanos imunocomprometidos. Os fatores sigma de função extracitoplasmática (ECF) são membros de sistemas de sinalização de superfície celular (CSS), abundantes em P. aeruginosa. Vinte genes codificando fatores sigma ECF estão presentes nos genomas sequenciados de P. aeruginosa, a maioria fazendo parte de sistemas TonB relacionados à captação de ferro. Neste trabalho, seis fatores sigma pobremente caracterizados foram superexpressos na linhagem PA14 a partir de um promotor induzível por arabinose para investigar seu papel na expressão dos sistemas de dois componentes PvrSR e RcsCB, que atuam na regulação da fímbria CupD, além de sua influência no crescimento de culturas de P. aeruginosa. Não foi observado efeito positivo de nenhum dos fatores sigma testados na expressão dos sistemas de dois componentes e a superexpressão de cinco deles tampouco levou a qualquer alteração no crescimento, porém a produção de piocianina foi alterada na superexpressão de PA14_55550 e a superexpressão de PA14_26600 e PA14_46810 levou a um discreto aumento no início da formação de biofilme em PA14. Por outro lado, culturas superexpressando σx (ALB04) apresentaram um perfil alterado de lipopolissacarídeo e uma curva de crescimento bifásica, alcançando precocemente uma fase estacionária seguida de uma recuperação do crescimento até uma segunda fase estacionária. Durante a primeira fase estacionária, a maior parte das células aumenta de tamanho e morre, mas as células remanescentes retornam à morfologia selvagem e seguem para a segunda fase de crescimento exponencial. Isso não acontece devido a mutações compensatórias, uma vez que células coletadas de pontos tardios da curva e diluídas em meio novo repetem este comportamento. Apesar de trabalhos com a linhagem PAO1 associarem σx à transcrição de oprF, que codifica a principal porina não específica de Pseudomonas, nas condições dos nossos ensaios em PA14 a expressão dessa porina não foi induzida pela superexpressão de σx. Assim, os efeitos observados nessa superexpressão também não podem ser atribuídos a OprF. A transcrição de oprF em PA14 mostrou-se majoritariamente dependente da região promotora a que se atribui a ligação de σ70, ao contrário dos relatos na literatura da dependência da região de ligação a σx. Análises proteômicas foram realizadas para investigar os elementos envolvidos nesses efeitos de superexpressão de σx, o que revelou a indução de diversas enzimas envolvidas na via de biossíntese de ácidos graxos. As células superexpressando σx apresentam uma maior proporção de ácidos hexadecanoico (C16) e hexadecenoico (C16:1) e dados de anisotropia mostram uma maior fluidez da(s) membrana(s). Este trabalho é o primeiro relato de um fator sigma ECF envolvido em biossíntese de lipídeos em P. aeruginosa. / Pseudomonas aeruginosa is a very versatile gammaproteobacteria, able to colonize different environments and to infect phylogenetically distinct hosts, including immunocompromised humans. The extracytoplasmic function sigma factors (ECFs) are members of cell signaling systems (CSS), abundant in P. aeruginosa. Twenty genes coding for ECF sigma factors are present in the sequenced genomes of P. aeruginosa, most of them being part of TonB systems related to iron uptake. In this work, six poorly characterized sigma factors were overexpressed in strain PA14 from an arabinose inducible promoter to investigate their role in the expression of the two-component systems PvrSR and RcsCB, which regulates CupD fimbria, and their influence in P. aeruginosa cultures growth. None of the tested sigma factors led to two-component systems upregulation and overexpression of five of them caused no change in the growth profile, but pyocyanin production was altered in PA14_55550 overexpression and PA14_26600 and PA14_46810 overexpression led to a slight increase in biofilm initiation in PA14. By the other side, cultures overexpressing σx (ALB04) presented an altered lipopolysaccharide profile and a biphasic growth curve, reaching an early stationary phase followed by a growth resuming untill a second stationary phase. During the early stationary phase, most cells swells and dies, but the remaining cells return to wild type morphology and proceed to the second exponential phase of growth. This is not due to compensatory mutations, since cells collected from late points of the curve and diluted in fresh medium repeat this behavior. Although studies with strain PAO1 associate σx with transcription of oprF, encoding the major nonspecific porin of Pseudomonas, under our experiments conditions with PA14, this porin expression is not induced by σx overexpression. Thus, the effects observed in this overexpression cannot be attributed to OprF. Transcription of oprF in PA14 proved to be mainly controlled by the σ70-dependent promoter region instead of the σx-dependent promoter region reported in the literature. Proteomic analyses were performed to investigate the elements involved in these effects of σx overexpression, which revealed the induction of several enzymes involved in fatty acids biosynthesis. Cells overexpressing σx exhibit a greater proportion of hexadecanoic (C16) and hexadecenoic (C16: 1) acids and anisotropy data show higher fluidity of the membrane (s). This work is the first report of an ECF sigma factor involved in lipid biosynthesis in P. aeruginosa.