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Desde 2008 o óleo diesel comercializado no território nacional tem recebido a adição crescente de biodiesel, além da redução gradativa nos teores de enxofre, que é uma tendência mundial de minimizar o impacto gerado pela queima de combustíveis fósseis. Aspectos relacionados com a qualidade final das misturas diesel com biodiesel, desde a produção até o armazenamento, tem recebido especial atenção. O objetivo desse trabalho foi avaliar a suscetibilidade à contaminação microbiana, durante o armazenamento simulado, do óleo diesel puro e mistura B10 com diferentes teores de enxofre: ULSD (S10), LSD (S50), HSD (S500) e UHSD (S1800). Os resultados dessa pesquisa mostraram que Pseudallescheria boydii apresentou maior habilidade de crescer na mistura B10 que Paecilomyces variotii. As análises de degradação, por espectroscopia no infravermelho, revelaram que os combustíveis avaliados possuem distintas características moleculares de acordo com os teores de enxofre. Os dados de RMN H-1 mostraram que o diesel com maior teor de enxofre (UHSD) apresentou reduzido teor de ramificações e compostos aromáticos comparado ao diesel com menor teor de enxofre (ULSD). O sequenciamento de alto rendimento (Illumina) do rDNA 16 e 18S mostrou que os Filos mais abundantes presentes no inóculo misto ao longo do tempo foram: Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Ascomycota e Basidiomycota. Os prováveis metabólitos identificados por UPLC/Q-ToF - MS, na fase aquosa, foram os ácidos: mirístico, palmítico, esteárico, oleico, linoleico e α-linolênico com abundância relativa de 2-10 vezes superior aos controles, indicando degradação de origem microbiana. Entre os 10 parâmetros avaliados de estabilidade química, somente a estabilidade oxidativa apresentou desconformidade com o Regulamento Técnico ANP e os níveis de contaminação microbiana, ao final de 90 dias, foram considerados aceitáveis. O conjunto de dados sugere que a redução de compostos sulfurados no diesel não induz suscetibilidade à contaminação microbiana, porém fatores como a origem da matéria prima, processo de refino e adição de biodiesel devem ser considerados. Os resultados desses estudos auxiliam na melhor compreensão dos microrganismos e metabólitos envolvidos nos processos de biocorrosão e biodeterioração do diesel e suas misturas visando a qualidade final do combustível. / Since 2008, the diesel fuel sold in the country has received the increasing addition of biodiesel, in addition to the gradual reduction in sulfur content, which is a global trend to minimize the impact generated by burning fossil fuels. Aspects related to the final quality of diesel and biodiesel blends, from production to storage, has received special attention. The aim of this study was to evaluate the susceptibility to microbial contamination during simulated storage, in pure diesel and blend B10 with different sulfur contents: ULSD (S10), LSD (S50), HSD (S500) and UHSD (S1800). The results of this research showed that Pseudallescheria boydii had greater ability to grow in the blend B10 than Paecilomyces variotii. The degradation analysis, infrared spectroscopy, showed that these fuels have different molecular characteristics according to their sulfur content. The data H1 NMR showed that the diesel with higher sulfur content (UHSD) showed a low level of branching and aromatic compounds compared to diesel fuel with lower sulfur content (ULSD). The high throughput sequencing (Illumina) of 16S and 18S rDNA showed the Phyla most abundant present in the mixed inoculum over time were: Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Ascomycota and Basidiomycota. Putative metabolites identified by UPLC / Q-ToF-MS in the water phase were: myristic, palmitic, stearic, oleic, linoleic and α-linolenic acids with relative abundance of 2-10 times higher than controls indicating degradation of microbial origin. Among the 10 evaluated parameters for chemical stability, only oxidative stability showed nonconformity with the ANP Technical Regulation and microbial contamination levels at the end of 90 days were considered acceptable. The data set suggest that the reduction of sulfur compounds in diesel fuel are not induced microbial susceptibility contamination, but factors such as the origin of the raw material, refining process and adding biodiesel to be considered. The results of these studies help to better understand the microorganisms and metabolites involved in the biocorrosion and biodeterioration processes of diesel and their blends aiming at the final quality of the fuel.

Den största akuta risken inom dricksvattenförsörjningen är vattenburen smittspridning. Dricksvattenrelaterad smittspridning förekommer i störst utsträckning i utvecklingsländer med dåliga sanitära förhållanden. Under 2000-talet har de nordiska länderna drabbats av flertalet dricksvattenburna sjukdomsutbrott vilket har lett till att mikrobiella föroreningar uppmärksammats som ett problem även i Norden. För att säkerställa att det dricksvatten som distribueras är av god mikrobiologisk kvalitet krävs att säkerhetsbarriärerna i vattenverket är tillräckligt effektiva för alla typer av mikrobiella föroreningar. Vissa mikroorganismer har uppmärksammats som mer tåliga mot olika desinfektionsmetoder, till exempel klorering, än vad som tidigare varit känt. För ett säkrare dricksvatten är det därför viktigt att öka kunskapen om mikroorganismers varierande motståndskraft mot olika desinfektionsmedel samt undersöka alternativ till de avskiljnings- och inaktiveringsmetoder som traditionellt används idag. Examensarbetets syfte var att studera de bakterier och virus som har störst relevans för dricksvattenkvaliteten och som tros kunna utgöra ett hot mot nordiskt dricksvatten, att undersöka orsakerna bakom dricksvattenburna utbrott samt att undersöka effektiviteten hos olika avskiljnings- och inaktiveringsmetoder för bakterier och virus. Detta gjordes genom en utförlig litteraturstudie samt en fallstudie. Fallstudiens syfte var att belysa problematiken inom dricksvattenberedning samt distribution i Norden genom verkliga exempel.  Studien visade att vattenburna sjukdomsutbrott normalt orsakas av kontaminering från avloppsvatten till dricksvatten. Detta sker antingen direkt till råvattnet, ofta till följd av kraftig nederbörd, eller under distributionen på grund av driftstörningar. Att kontamineringen tillåts nå konsumenten är ett resultat av inadekvata reningsmetoder i vattenverken. Problematiken inom dricksvattenområdet beror bland annat på användandet av bristfälliga indikatororganismer vilka är mindre tåliga och har sämre överlevnadsförmåga än många sjukdomsalstrande mikroorganismer. Tidskrävande och osäkra analysmetoder utgör också ett stort problem. Svårigheter i att identifiera den orsakande mikroorganismen samt att bekräfta utbrottet som dricksvattenrelaterat har lett till att mörkertalet gällande antalet vattenburna sjukdomsfall är stort. Det finns ett stort behov av ökad kunskap gällande olika mikroorganismers tålighet mot desinfektion samt ett behov av reglering för kontroll av råvattenkvaliteten och den mikrobiologiska säkerheten. Utifrån studien kunde även konstateras att antalet vattenburna utbrott tros öka med kraftigare nederbörd och extremväder till följd av klimatförändringarna. / Waterborne diseases are considered a major problem in the water supply. Spreading of infection through drinking water occurs predominantly in developing countries with poor sanitation. During the 21th century the Nordic countries have been affected by several waterborne outbreaks associated with drinking water. As a result, microbial contamination of drinking water has been recognized as a problem in the Nordic countries as well. To ensure high quality drinking water it is required that the hygienic barriers used in the water treatment plant are efficient in reducing all types of microbial contaminants. Some microorganisms have shown a greater resistance against commonly used disinfectants then previously known. To secure the distribution of safe drinking water it is essential to increase the knowledge regarding the varying resistance of microorganisms and to investigate alternative water treatment processes to those traditionally used in drinking water treatment today.  The objective to this master thesis was to study the bacteria and viruses that are most relevant for drinking water quality and may compose a threat to the drinking water quality in the Nordic countries, to investigate the cause of waterborne outbreaks and to study the effectiveness in reducing bacteria and viruses for various water treatment processes. The study was performed through a thorough literature review and a case study. The purpose of the case study was to illustrate the issues in drinking water treatment and distribution in the Nordic region. The study showed that the majority of the outbreaks are caused by contamination from sewage water to drinking water. The contamination is allowed to reach the consumer as a result of inadequate treatment in the water works. Usage of inappropriate indicator organisms that are less resistant to disinfectants than a lot of pathogens are a big problem in drinking water treatment. Time-consuming and uncertain analytical methods also compose a great issue. Due to difficulties in identifying the causative microorganism and verify the outbreak as drinking water related it is believed that there is a large number of unrecorded cases of illness. There is a great need of increased understanding of the varying resistance of microorganisms to disinfectants and a need for regulation for monitoring raw water quality and microbiological safety. Based on the study it was also found that waterborne outbreaks are believed to increase as a result of extreme weather caused by climate change.

Klimatscenarier framtagna av SMHI visar bland annat på att nederbördsmängden kommer att öka i stora delar av Sverige. En ökad nederbördsmängd leder till att flöden i vattendrag och spillvattenledningar ökar, som i sin tur kan leda till att oönskade ämnen förorenar viktiga naturresurser så som dricksvattentäkter. Den största risken för mikrobiella föroreningar i vattentäkter och vattendrag kommer från Sveriges reningsverk, främst till följd av att reningsverkens flödeskapacitet överstigs på grund av kraftiga regn. Vid kraftiga regn riskerar reningsverk att bräddas, vilket leder till att ofullständigt renat spillvatten leds ut till sjöar.   Bålsta reningsverk, Håbo kommun, bräddas ibland på grund av höga flöden. Det otillräckligt renade spillvattnet leds då ut till reningsverkets utsläppspunkt söder om Bålsta stad i Norra Björkfjärden. Mindre än två kilometer ifrån reningsverkets utsläppspunkt ligger Bålsta vattenverks råvattenintag. Det är således av intresse att undersöka om vattenverkets råvattenintag påverkas av en bräddning. Utöver vattenverket ligger flera populära badplatser i närheten av reningsverkets utsläppspunkt.   Syftet med examensarbetet var att studera hur en bräddning av orenat spillvatten sprids i Mälaren under sommarens badsäsong, samt att identifiera vid vilka vind- och flödesförhållanden badplatserna och råvattentäkten har störst risk att förorenas. Syftet var även att studera hur en ökad nederbördsmängd påverkar delar av Bålsta reningsverks spillvattenledningsnät. För att nå examensarbetets syfte användes spridningsmodellen Delft3D samt ledningsnätmodellen MIKE-Urban. På grund av otillräcklig data om bland annat Bålsta reningsverks spillvattenledningsnät erhölls inget tillförlitligt resultat från MIKE-Urban modelleringen. Istället sammanställdes de saknade data om reningsverkets spillvattenledningsnät vilket resulterade i förslag om kompletterande åtgärder för att underlätta kommande spillvattenmodelleringar.    Resultat från spridningssimuleringen tydde på att ingen av de studerade platserna överskrider värden för utmärkt badvattenkvalitet enligt EU:s badvattendirektiv vid medelvindhastighet samt ett utsläpp av 77 m3 orenat spillvatten. En bräddning av 77 m3 orenat spillvatten baserades på reningsverkets bräddningsvolymer från år 2010. För badplatserna ansågs det fördelaktigt att en bräddning sker under vindstilla förhållanden till följd av begränsad omblandning mellan Mälarens temperaturskiktning. Vid ett två dygn långt utsläpp av totalt 5 000 m3 orenat spillvatten med fem sekundmeters vind erhöll majoriteten av badplatserna för höga patogenvärden. Reningsverkets personal bör i sådana situationer gå ut med allmänna varningar för badande i Norra Björkfjärden. / Climate change scenarios conducted by SMHI indicate increasing precipitation in most parts of Sweden. Such increases will lead to bigger flows in rivers and sewer networks, which in turn may lead to leakage of unwanted substances such as contagious microorganisms to drinking water supplies. The single largest risk of lake contamination is from sewage water treatment plants, mostly due to overflow of wastewater. High precipitation may flood the treatment plant causing untreated water to leak out and into surrounding lakes.   Bålsta treatment plant, in Håbo municipality, has overflow issues in periods with high precipitation, which lead to leakage of inadequately treated wastewater and thus local contamination in Lake Mälaren. Less than two kilometers from the release point of Bålsta treatment plant, Bålsta drinking water plant has its intake of raw water. Hence it is of great interest to study the dispersal of Bålsta treatment plant’s wastewater in Lake Mälaren to prevent unwanted human exposure to contagious microorganisms. Håbo municipality also has popular swimming spots close to the treatment plant’s outlet.   The aim of this study is to examine the pathogenic impact that an overflow of Bålsta treatment plant in summer time has on Bålsta drinking water plant and bathing spots. The study hoped to identify the conditions where certain places of interest were to be contaminated. In addition, the aim was to investigate the changes of sewage flow due to an increased precipitation. Two models were used to reach the aforementioned aims: (1.) a hydrodynamic model (Delft3D) to study the spread of wastewater following an overflow and (2.) an urban water model (MIKE–Urban) to study the total amount of sewage flow.   Due to insufficient data concerning Bålsta’s sewage system, simulations from MIKE–Urban were unable to provide reliable results. Large parts of the sewage network had missing data regarding pipe dimensions, positioning and material, which resulted in a model with little precision. Instead of providing unreliable results, the study resulted in recommendations for further data collecting to help future investigations of Bålsta’s sewage system.   With low overflows of untreated water, Delft3D modeling showed that none of the places of interest had pathogen levels that exceeded the European Union’s Bathwater quality directive when simulated with mean values of wind, treated wastewater flow and wind direction. When extreme amounts of overflow were simulated in strong winds, five of seven places of interest were unfit for bathing according to the directive. In case of an overflow of more than the simulated outflow of 77 m3 untreated wastewater, the operators of Bålsta’s treatment plant need to perform actions to prevent human exposure.

Thesis (MSc Food Sc (Food Science))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: Over the last decade, studies on the quality of the water in many of South Africa.s rivers revealed an increase in pollution levels (DWAF, 1996; DWAF, 2000). In agricultural areas of the Western Cape many farmers draw water for irrigation from nearby rivers without knowing what the microbial pollution level is. In order to investigate the current quality of river water in some of the Western Cape.s major rivers and to explore the possibility of pathogen carryover from water onto produce, this research project was initiated. In an exploratory study over a five month period, the microbiological and water chemistry of three selected sites from the upper Berg and two from the Plankenbrug Rivers were assessed. Studied organisms included Salmonella, Staphylococcus, Listeria, endosporeformers, coliforms, Escherichia coli and intestinal Enterococci, while the chemical analysis consisted of pH, alkalinity, conductivity and chemical oxygen demand (COD). Faecal coliform counts ranging from 540 to 1 700 000 cfu.100ml-1 and 490 to 160 000 cfu.100ml-1 were found for the Berg and Plankenbrug Rivers, respectively. The water temperature ranged from 12º to 21ºC with COD values always below 100 mg.L-1. Potential human pathogens such as Salmonella, Staphylococcus, Listeria, endosporeformers, E. coli and intestinal Enterococci were frequently isolated from all five sites that were sampled. These results are of great concern to farmers, fresh produce retailers and consumers alike as the river water is regularly drawn for irrigation of produce that is later consumed raw or after a minimal processing step. From the exploratory study it was concluded that the water from all the sites were not suitable for use in irrigation practices as they regularly exceeded the guidelines for faecal coliforms and E.coli as set out by South African authorities. Irrigation with faecally polluted river water is one way that fruit and vegetables can become contaminated with foodborne pathogens. The risk of disease transmission from potential pathogens present in the irrigation water is influenced by the microbial load present and the numbers carried over to the produce. In this study the carry-over and survival of Escherichia coli on green beans, sugar-snap peas and cocktail tomatoes was assessed under controlled laboratory conditions. The produce was exposed to E.coli under different combinations of exposure times (5, 15 or 30 min), drying times (30 or 120 min) and different inoculum concentration ranges (107, 105, 104, 10³ and 10ª). In all cases a reduction of at least one log value in original inoculum number was found with the 10ª inoculums showing no survivors. Follow-up studies with an exposure time of 60 min and increased drying times of 6 or 12 h. Neither of these parameter changes affected the variation in numbers for the same inoculum or the E.coli survivors. Similar E.coli loads to those detected on the green beans were detected on the sugar-snap peas. The number of survivors on the cocktail tomatoes was much lower than found for the beans and peas. The number of survivors on the cocktail tomatoes was much lower than found for the beans and peas. This was attributed the 'smooth'surface of the tomato skin probably making attachment of the E.coli bacteria difficult. With the exception of the 10ª inoculum range, all E.coli survivors detected on the three types of produce studied exceeded the guideline numbers set for fresh produce. If similar survival patterns are to be found in the environment then results from this study should serve as a warning that the Plankenburg river water is unsafe for use in the irrigation of fresh produce. Some farmers are already treating river water with chlorine prior to irrigation to eliminate the chance of pathogen transfer onto produce. However, this is not a feasible solution for most farmers due to the high cost of implementing a system such as this. / AFRIKAANSE OPSOMMING: Oor die afgelope tien jaar het studies oor die gehalte van die water in baie van Suid-Afrika se riviere 'n toename in besoedelingsvlakke getoon. Baie boere in landbou gebiede van die Wes-Kaap wat watertrek vir besproeiing uit die nabygelee riviere is onbewus van die mikrobiese besoedelingsvlak van die rivier. Hierdie navorsingsprojek is geinisieer ten einde die huidige gehalte van die water in sommige van die Wes-Kaap se riviere te monitor, asook die moontlikheid van patogeen oordrag van die water na vars produkte te ondersoek. Met behulp van 'n verkennende studie oor . vyf maande tydperk, is die mikrobiologiese en water chemie kenmerke van drie geselekteerde moniteringspunte uit die boonste Bergrivier, en twee uit die Plankenburgrivier bemonster. Die bestudeerde organismes het ingesluit: Salmonella, Staphylococcus, Listeria, endosporeformers, fekale kolivorme, Escherichia coli en intestinale Enterococci. Die chemiese parameters het bestaan uit pH, alkaliniteit, geleiding, temperatuur en chemiese suurstof behoeftes (CSB). Fekale kolivorme tellings het gewissel van 540 - 1 700 000 cfu.100ml-1 en 490 tot 160 000 cfu.100ml-1 is gevind vir die Berg- en Plankenburgriviere, onderskeidelik. Die water temperatuur het gewissel van 12 º tot 21ºC met die CSB-waarde altyd onder 100 mg.L-1. Potensiele menslike patogene soos Salmonella, Staphylococcus, Listeria, endosporeformers, E. coli en intestinale Enterococci is dikwels geisoleer uit al vyf die moniteringspunte. Hierdie resultate is van groot belang vir boere, kleinhandelaars en varsprodukte verbruikers omdat die riviere se water gereeld onttrek word vir besproeiing van produkte wat later gebruik word in rou of in minimale geprosesseerde vorm. Vanuit die verkennende en uit die langer termyn studie (12 maande) is die gevolgtrekking gemaak dat die water vanuit die riviere nie geskik is vir gebruik in besproeiingspraktyke waar die riglyne vir fekale kolivorme en E. coli, soos uiteengesit deur die Suid-Afrikaanse owerhede, gereeld oorskry word nie. Besproeiing met fekale besoedelde rivierwater is een manier waarop groente en vrugte besmet kan word met voedsel patogene. Die risiko vir die oordrag van moontlike patogene teenwoordig in die besproeiingswater word beinvloed deur die mikrobiese lading teenwoordig en die getalle wat oorgedra word na die produkte. In hierdie studie is die oordrag en oorlewing van E. coli op groenboontjies, ertjies en tamaties onder beheerde laboratoriumtoestande beoordeel. Die produkte is blootgestel aan E. coli onder verskillende kombinasies van blootstellingstydperke (5, 15 of 30 min), drogingstye (30 of 120 min) en verskillende inokulum konsentrasies (107, 105, 104, 10³ en 10ª). In elke geval is . afname van ten minste een log waarde van die oorspronklike inokulum konsentrasie gevind. Die 10ª inokulum reeks het geen oorlewende selle getoon nie. Opvolg studies is gedoen met 'n blootstellingstyd van 60 min en verlengde drogingstye van 6 of 12 uur. Daar is gevind dat hierdie parameter veranderings die variasie in getalle vir dieselfde inokulum of die oorlewende E. coli nie beinvloed het nie. Soortgelyke E. coli ladings wat op die groenbone aangetref is, is ook op die ertjies aangetref. Die aantal oorlewendes op die tamaties was heelwat laer as die resultate vir die boontjies en ertjies. Dit word toegeskryf aan die "gladde" oppervlak van die tamatie-vel wat waarskynlik die aanhegting van die E. coli selle bemoeilik. Met die uitsondering van die 10ª inokulum reeks, het al die E. coli oorlewende selle gevind op die drie tipes produkte, die riglyn getalle oorskry wat vir vars produkte gestel is. As soortgelyke oorlewingspatrone gevind word in die omgewing, behoort die resultate van hierdie studie as waarskuwing te dien dat die water van die Plankenburgrivier ongeskik is vir die besproeiing van vars produkte. Sommige boere is reeds besig met chloor behandeling van die rivierwater voor besproeiing, om die kans van patogeenoordrag na die produkte uit te skakel. Dit is egter nie 'n haalbare oplossing vir die meeste boere nie as gevolg van die hoe kostes verbonde aan die implementering van' n stelsel soos hierdie.

Desde 2008 o óleo diesel comercializado no território nacional tem recebido a adição crescente de biodiesel, além da redução gradativa nos teores de enxofre, que é uma tendência mundial de minimizar o impacto gerado pela queima de combustíveis fósseis. Aspectos relacionados com a qualidade final das misturas diesel com biodiesel, desde a produção até o armazenamento, tem recebido especial atenção. O objetivo desse trabalho foi avaliar a suscetibilidade à contaminação microbiana, durante o armazenamento simulado, do óleo diesel puro e mistura B10 com diferentes teores de enxofre: ULSD (S10), LSD (S50), HSD (S500) e UHSD (S1800). Os resultados dessa pesquisa mostraram que Pseudallescheria boydii apresentou maior habilidade de crescer na mistura B10 que Paecilomyces variotii. As análises de degradação, por espectroscopia no infravermelho, revelaram que os combustíveis avaliados possuem distintas características moleculares de acordo com os teores de enxofre. Os dados de RMN H-1 mostraram que o diesel com maior teor de enxofre (UHSD) apresentou reduzido teor de ramificações e compostos aromáticos comparado ao diesel com menor teor de enxofre (ULSD). O sequenciamento de alto rendimento (Illumina) do rDNA 16 e 18S mostrou que os Filos mais abundantes presentes no inóculo misto ao longo do tempo foram: Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Ascomycota e Basidiomycota. Os prováveis metabólitos identificados por UPLC/Q-ToF - MS, na fase aquosa, foram os ácidos: mirístico, palmítico, esteárico, oleico, linoleico e α-linolênico com abundância relativa de 2-10 vezes superior aos controles, indicando degradação de origem microbiana. Entre os 10 parâmetros avaliados de estabilidade química, somente a estabilidade oxidativa apresentou desconformidade com o Regulamento Técnico ANP e os níveis de contaminação microbiana, ao final de 90 dias, foram considerados aceitáveis. O conjunto de dados sugere que a redução de compostos sulfurados no diesel não induz suscetibilidade à contaminação microbiana, porém fatores como a origem da matéria prima, processo de refino e adição de biodiesel devem ser considerados. Os resultados desses estudos auxiliam na melhor compreensão dos microrganismos e metabólitos envolvidos nos processos de biocorrosão e biodeterioração do diesel e suas misturas visando a qualidade final do combustível. / Since 2008, the diesel fuel sold in the country has received the increasing addition of biodiesel, in addition to the gradual reduction in sulfur content, which is a global trend to minimize the impact generated by burning fossil fuels. Aspects related to the final quality of diesel and biodiesel blends, from production to storage, has received special attention. The aim of this study was to evaluate the susceptibility to microbial contamination during simulated storage, in pure diesel and blend B10 with different sulfur contents: ULSD (S10), LSD (S50), HSD (S500) and UHSD (S1800). The results of this research showed that Pseudallescheria boydii had greater ability to grow in the blend B10 than Paecilomyces variotii. The degradation analysis, infrared spectroscopy, showed that these fuels have different molecular characteristics according to their sulfur content. The data H1 NMR showed that the diesel with higher sulfur content (UHSD) showed a low level of branching and aromatic compounds compared to diesel fuel with lower sulfur content (ULSD). The high throughput sequencing (Illumina) of 16S and 18S rDNA showed the Phyla most abundant present in the mixed inoculum over time were: Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Ascomycota and Basidiomycota. Putative metabolites identified by UPLC / Q-ToF-MS in the water phase were: myristic, palmitic, stearic, oleic, linoleic and α-linolenic acids with relative abundance of 2-10 times higher than controls indicating degradation of microbial origin. Among the 10 evaluated parameters for chemical stability, only oxidative stability showed nonconformity with the ANP Technical Regulation and microbial contamination levels at the end of 90 days were considered acceptable. The data set suggest that the reduction of sulfur compounds in diesel fuel are not induced microbial susceptibility contamination, but factors such as the origin of the raw material, refining process and adding biodiesel to be considered. The results of these studies help to better understand the microorganisms and metabolites involved in the biocorrosion and biodeterioration processes of diesel and their blends aiming at the final quality of the fuel.

Fundação Araucária / Fluidos de corte são utilizados para auxiliar no processo de usinagem, tendo por finalidade a refrigeração. Além disso, promove a lubrificação, a diminuição do atrito em relação à peça usinada e a ferramenta de corte, aumentando assim a eficiência do trabalho das fábricas. No sistema de usinagem, pode ocorrer o crescimento de microrganismos, uma vez que estes utilizam o fluido de corte como fonte de alimento, reduzindo assim a vida útil dos fluidos, proporcionando perdas econômicas e causando danos à saúde do operador da máquina. Para minimizar a presença de microrganismos, são utilizados produtos químicos denominados biocidas, que em altas concentrações pode causar danos à saúde. Dentro desse panorama, o objetivo desta pesquisa foi avaliar a eficiência dos óleos essenciais de Ocimum gratissimum (alfavacão), Cymbopogon winterianus (citronela) Syzygium aromaticum (cravo) e Carapa guianensis (andiroba) como agentes de controle do crescimento de bactérias em substituição ao biocida no sistema de usinagem. Inicialmente, foram realizadas coletas de fluido de corte em uma indústria automobilística para quantificar, isolar e caracterizar as bactérias (heterotróficas e potencialmente patogênicas) presentes no fluido de corte. Na sequência, as bactérias isoladas foram testadas quanto a susceptibilidade às diferentes concentrações dos óleos essenciais. O estudo procurou definir a concentração mínima inibitória (CMI) e antibiograma para avaliar a resistência a antibióticos das bactérias isoladas. Realizou-se, então, o teste de tempo de contato dos óleos essenciais e de encapsulamento do óleo essencial de citronela por meio da técnica de gelificação iônica, que consistiu em simular a situação do reservatório (i) com óleo encapsulado, (ii) com óleo misturado diretamente ao fluido de corte (iii) com fluido de corte contaminado, com o objetivo de avaliar a melhor forma de aplicação dos óleos no sistema de usinagem. Dentre os resultados, obteve-se 1,63x107 e 2,2x107 UFC/mL de bactérias heterotróficas e 3,5x105 e 2x106 UFC/mL de bactérias potencialmente patogênicas, na primeira e segunda coleta de fluidos, respectivamente. Foram isolados seis grupos de bactérias correspondentes às seguintes espécies: Staphylococcus aureus, Enterobacter cloacae, Escherichia coli, Enterococcus faecalis, Proteus mirabilis e Acinetobacter spp. A bactéria que apresentou maior resistência aos antibióticos testados foi a Acinetobacter spp As que apresentaram maior sensibilidade foram a Enterobacter cloacae (A2), Escherichia coli (A3) e Proteus mirabilis (A6). O óleo de Syzygium aromaticum teve o maior halo formado na menor concentração testada (0,25%) - 11,07 mm, seguido por Ocimum gratissimum, com halo de 11,0 mm, e Cymbopogon winterianus ,10,7 mm. No teste de tempo de contato, o menor tempo de redução de crescimento ocorreu em 1 minuto para Ocimum gratissimum; 5 minutos para Syzygium aromaticum e 10 minutos para Cymbopogon nardus. O teste de encapsulamento foi realizado com o óleo de Cymbopogon nardus, obtendo-se redução significativa nas duas formas de aplicação do óleo essencial. / Cutting fluids are used to aid in the machining process, with the purpose of cooling, besides promoting lubrication, reducing friction in relation to the machined part and the cutting tool, thus increasing the efficiency of the work of the factories.In the machining system, the growth of microorganisms can occur, since they use the cutting fluid as food source, thus reducing the useful life of the fluids and providing economic losses and causing damages to the health of the worker.To minimize the presence of microorganisms is used a chemical called biocide, which in high concentrations can cause health damage.The objective of this research was to evaluate the efficiency of the essential oils of Ocimum gratissimum (Alfavaca), Cymbopogon nardus (Citronella) Syzygium aromaticum (Clove) and Carapa guianensis (andiroba) as a control agent for the growth of bacteria in replacement of the biocide in the machining system.Initially, cutting fluid samples were collected in an automobile industry to quantify, isolate and characterize the bacteria (heterotrophic and potentially pathogenic) present in the cutting fluid.Following the isolated bacteria were tested for susceptibility to the different concentrations of the oils to define the minimum inhibitory concentration (MIC) and antibiogram to evaluate the resistance of the bacteria isolated to antibiotics.The contact time test of the essential oils and encapsulation of the essential oil of Citronella was carried out by means of the ionic gelation technique, which consisted of the simulation of reservoirs containing cut fluid contaminated with encapsulated oil, with oil mixed directly with the oil. Cutting fluid and reservoir containing only contaminated cutting fluid, in order to evaluate the best way of applying the oils in the machining system.Among the results obtained were 1.63x107 and 2.2x107 CFU / mL heterotrophic bacteria and 3.5x105 and 2x106 CFU / mL of potentially pathogenic bacteria in the first and second fluids collection, respectively. Six groups of bacteria were isolated, corresponding to the following species: Staphylococcus aureus, Enterobacter cloacae, Escherichia coli, Enterococcus faecalis, Proteus mirabilis and Acinetobacter spp.The bacterium that presented the highest resistance to the antibiotics tested was Acinetobacter spp and the ones with the highest sensitivity were Enterobacter cloacae (A2), Escherichia coli (A3) and Proteus mirabilis (A6). The oil of Syzygium aromaticum had the largest halo formed by the lowest concentration tested (0.25%) -11.07 mm, followed by Ocimum gratissimum with halo of 11 mm and Cymbopogon nardus 10.7 mm. In the test of contact time the shortest time reduction occurred in 1 minute for Ocimum gratissimum, 5 minutes for Syzygium aromaticum and 10 minutes Cymbopogon nardus for 4 of 6 bacterial isolates. The encapsulation test was performed with the Cymbopogon nardus oil, obtaining a significant reduction in the two forms of essential oil application.

O fluido de corte é largamente utilizado no processo de usinagem de metais, porém é vulnerável à contaminação microbiológica que contribuem para sua perda de qualidade fazendo com que diminua o seu tempo de vida útil. Tendo em vista que diversos trabalhos estudam a contaminação microbiológica do fluido, mas poucos relacionam com as alterações químicas do óleo constituinte do fluido, o objetivo do presente trabalho foi monitorar o crescimento de microrganismos em duas bases de fluidos de corte cada um contendo um tipo de óleo (mineral e vegetal) sob duas condições de funcionamento de máquina ferramenta: processo normal e processo utilizando um catalisador eletrofísico que direcionava fluxo de ozônio no fluido de corte, avaliar as mudanças químicas do óleo extraído do fluido e a capacidade dos microrganismos isolados de degradar o fluido de corte e o óleo extraído. Foram realizados contagem e isolamento de bactérias e fungos no fluido de corte utilizado na máquina ferramenta ao longo de 166 horas de utilização da máquina. Para a análise da qualidade do óleo, este foi extraído do fluido de corte e posteriormente medidos o índice de acidez (óleo mineral e vegetal), iodo e saponificação do óleo vegetal. Para o teste de degradação de hidrocarbonetos foi utilizado o 2,6 diclorofenol-indofenol como indicador de degradação, sendo as fontes de carbono: fluido de corte e os óleos extraído do fluido de corte em diferentes tempos de uso da máquina. Os resultados demonstraram que no fluido de corte mineral operado na máquina por 126 horas a quantidade de bactérias e fungos foi na ordem de 105 e 102 UFC/mL, respectivamente, com a utilização do catalisador (em 40 horas de operação da máquina) foi para 106 e 101 UFC/mL. No fluido de corte vegetal os valores foram de 102 UFC/mL para bactérias e fungos e após a aplicação do catalisador as quantidades foram para 105 e 103 UFC/mL, respectivamente. Dos microrganismos isolados do fluido de corte mineral que foram avaliados quanto a degradação de hidrocarbonetos, verificou-se que 42% das bactérias e 84% dos fungos foram capazes de degradar o fluido de corte ou o óleo em 5 dias de análise. Enquanto 54% das bactérias e 74% dos fungos do fluido de corte vegetal foram capazes de degradar o fluido de corte ou o óleo. Pode-se concluir que houve alterações químicas no óleo utilizado no fluido de corte e que os microrganismos podem ter influenciado diretamente na degradação do óleo, pois o aumento da quantidade de microrganismos acompanhou a degradação do óleo. / The cutting fluid is widely used in metal machining process, but it is vulnerable to microbiological contamination contributing to loss in quality causing the decrease of its useful lifetime. Considering that many papers study the microbiological contamination of the fluid, but few associated to chemical changes of the oil constituent of the fluid, the objective of this study was to monitor the growth of microorganisms in two cutting fluids bases each containing a type of oil (mineral and vegetable) under two operating conditions of the machine tool: normal process and process using a electrophysical catalyst which directs the flow of ozone to the cutting fluid, to evaluate the chemical changes of the fluid oil extracted and the capability of microorganisms to degrade the cutting fluid and the oil extracted. Counting and isolation of bacteria and fungi were performed on cutting fluids used in machine tool throughout 166 hours of operation of the machine. For analysis of oil quality, it was extracted oil of the cutting fluid and posteriorly it was measured the acidity index (mineral and vegetable oils), iodine and saponification of vegetable oil. For the hydrocarbon degradation test it was used 2,6-dichlorophenol-indophenol as degradation indicator, being carbon sources: cutting fluid and oil extracted from the cutting fluid at different times of use of the machine. The results demonstrated that the amount of bacteria and fungi present in the mineral cutting fluid operated on the machine for 126 hours was in the order of 105 to 102 CFU/ml, respectively, and using the catalyst (40 hours of machine operation) it was 106 and 101 CFU/mL, respectively. The amount of bacteria and fungi from vegetable cutting fluid were 102 CFU/ml and after the application of the catalyst they were 105 and 103 CFU/ml, respectively. In the degradation tests of hydrocarbon, it was found that 42% of bacteria and 84% of the fungi were able to degrade the mineral cutting fluid or extracted oil on 5 days of analysis. 54% of bacteria and 74% of the fungi found on vegetable cutting fluid were able to degrade the cutting fluid or extracted oil. It can be concluded that there were chemical changes in the oils used to prepare cutting fluid and that microorganisms can be directly influenced in oil degradation, since the increased amount of microorganisms followed the oil degradation.

Fundação Araucária / Fluidos de corte são utilizados para auxiliar no processo de usinagem, tendo por finalidade a refrigeração. Além disso, promove a lubrificação, a diminuição do atrito em relação à peça usinada e a ferramenta de corte, aumentando assim a eficiência do trabalho das fábricas. No sistema de usinagem, pode ocorrer o crescimento de microrganismos, uma vez que estes utilizam o fluido de corte como fonte de alimento, reduzindo assim a vida útil dos fluidos, proporcionando perdas econômicas e causando danos à saúde do operador da máquina. Para minimizar a presença de microrganismos, são utilizados produtos químicos denominados biocidas, que em altas concentrações pode causar danos à saúde. Dentro desse panorama, o objetivo desta pesquisa foi avaliar a eficiência dos óleos essenciais de Ocimum gratissimum (alfavacão), Cymbopogon winterianus (citronela) Syzygium aromaticum (cravo) e Carapa guianensis (andiroba) como agentes de controle do crescimento de bactérias em substituição ao biocida no sistema de usinagem. Inicialmente, foram realizadas coletas de fluido de corte em uma indústria automobilística para quantificar, isolar e caracterizar as bactérias (heterotróficas e potencialmente patogênicas) presentes no fluido de corte. Na sequência, as bactérias isoladas foram testadas quanto a susceptibilidade às diferentes concentrações dos óleos essenciais. O estudo procurou definir a concentração mínima inibitória (CMI) e antibiograma para avaliar a resistência a antibióticos das bactérias isoladas. Realizou-se, então, o teste de tempo de contato dos óleos essenciais e de encapsulamento do óleo essencial de citronela por meio da técnica de gelificação iônica, que consistiu em simular a situação do reservatório (i) com óleo encapsulado, (ii) com óleo misturado diretamente ao fluido de corte (iii) com fluido de corte contaminado, com o objetivo de avaliar a melhor forma de aplicação dos óleos no sistema de usinagem. Dentre os resultados, obteve-se 1,63x107 e 2,2x107 UFC/mL de bactérias heterotróficas e 3,5x105 e 2x106 UFC/mL de bactérias potencialmente patogênicas, na primeira e segunda coleta de fluidos, respectivamente. Foram isolados seis grupos de bactérias correspondentes às seguintes espécies: Staphylococcus aureus, Enterobacter cloacae, Escherichia coli, Enterococcus faecalis, Proteus mirabilis e Acinetobacter spp. A bactéria que apresentou maior resistência aos antibióticos testados foi a Acinetobacter spp As que apresentaram maior sensibilidade foram a Enterobacter cloacae (A2), Escherichia coli (A3) e Proteus mirabilis (A6). O óleo de Syzygium aromaticum teve o maior halo formado na menor concentração testada (0,25%) - 11,07 mm, seguido por Ocimum gratissimum, com halo de 11,0 mm, e Cymbopogon winterianus ,10,7 mm. No teste de tempo de contato, o menor tempo de redução de crescimento ocorreu em 1 minuto para Ocimum gratissimum; 5 minutos para Syzygium aromaticum e 10 minutos para Cymbopogon nardus. O teste de encapsulamento foi realizado com o óleo de Cymbopogon nardus, obtendo-se redução significativa nas duas formas de aplicação do óleo essencial. / Cutting fluids are used to aid in the machining process, with the purpose of cooling, besides promoting lubrication, reducing friction in relation to the machined part and the cutting tool, thus increasing the efficiency of the work of the factories.In the machining system, the growth of microorganisms can occur, since they use the cutting fluid as food source, thus reducing the useful life of the fluids and providing economic losses and causing damages to the health of the worker.To minimize the presence of microorganisms is used a chemical called biocide, which in high concentrations can cause health damage.The objective of this research was to evaluate the efficiency of the essential oils of Ocimum gratissimum (Alfavaca), Cymbopogon nardus (Citronella) Syzygium aromaticum (Clove) and Carapa guianensis (andiroba) as a control agent for the growth of bacteria in replacement of the biocide in the machining system.Initially, cutting fluid samples were collected in an automobile industry to quantify, isolate and characterize the bacteria (heterotrophic and potentially pathogenic) present in the cutting fluid.Following the isolated bacteria were tested for susceptibility to the different concentrations of the oils to define the minimum inhibitory concentration (MIC) and antibiogram to evaluate the resistance of the bacteria isolated to antibiotics.The contact time test of the essential oils and encapsulation of the essential oil of Citronella was carried out by means of the ionic gelation technique, which consisted of the simulation of reservoirs containing cut fluid contaminated with encapsulated oil, with oil mixed directly with the oil. Cutting fluid and reservoir containing only contaminated cutting fluid, in order to evaluate the best way of applying the oils in the machining system.Among the results obtained were 1.63x107 and 2.2x107 CFU / mL heterotrophic bacteria and 3.5x105 and 2x106 CFU / mL of potentially pathogenic bacteria in the first and second fluids collection, respectively. Six groups of bacteria were isolated, corresponding to the following species: Staphylococcus aureus, Enterobacter cloacae, Escherichia coli, Enterococcus faecalis, Proteus mirabilis and Acinetobacter spp.The bacterium that presented the highest resistance to the antibiotics tested was Acinetobacter spp and the ones with the highest sensitivity were Enterobacter cloacae (A2), Escherichia coli (A3) and Proteus mirabilis (A6). The oil of Syzygium aromaticum had the largest halo formed by the lowest concentration tested (0.25%) -11.07 mm, followed by Ocimum gratissimum with halo of 11 mm and Cymbopogon nardus 10.7 mm. In the test of contact time the shortest time reduction occurred in 1 minute for Ocimum gratissimum, 5 minutes for Syzygium aromaticum and 10 minutes Cymbopogon nardus for 4 of 6 bacterial isolates. The encapsulation test was performed with the Cymbopogon nardus oil, obtaining a significant reduction in the two forms of essential oil application.

Desde 2008 o óleo diesel comercializado no território nacional tem recebido a adição crescente de biodiesel, além da redução gradativa nos teores de enxofre, que é uma tendência mundial de minimizar o impacto gerado pela queima de combustíveis fósseis. Aspectos relacionados com a qualidade final das misturas diesel com biodiesel, desde a produção até o armazenamento, tem recebido especial atenção. O objetivo desse trabalho foi avaliar a suscetibilidade à contaminação microbiana, durante o armazenamento simulado, do óleo diesel puro e mistura B10 com diferentes teores de enxofre: ULSD (S10), LSD (S50), HSD (S500) e UHSD (S1800). Os resultados dessa pesquisa mostraram que Pseudallescheria boydii apresentou maior habilidade de crescer na mistura B10 que Paecilomyces variotii. As análises de degradação, por espectroscopia no infravermelho, revelaram que os combustíveis avaliados possuem distintas características moleculares de acordo com os teores de enxofre. Os dados de RMN H-1 mostraram que o diesel com maior teor de enxofre (UHSD) apresentou reduzido teor de ramificações e compostos aromáticos comparado ao diesel com menor teor de enxofre (ULSD). O sequenciamento de alto rendimento (Illumina) do rDNA 16 e 18S mostrou que os Filos mais abundantes presentes no inóculo misto ao longo do tempo foram: Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Ascomycota e Basidiomycota. Os prováveis metabólitos identificados por UPLC/Q-ToF - MS, na fase aquosa, foram os ácidos: mirístico, palmítico, esteárico, oleico, linoleico e α-linolênico com abundância relativa de 2-10 vezes superior aos controles, indicando degradação de origem microbiana. Entre os 10 parâmetros avaliados de estabilidade química, somente a estabilidade oxidativa apresentou desconformidade com o Regulamento Técnico ANP e os níveis de contaminação microbiana, ao final de 90 dias, foram considerados aceitáveis. O conjunto de dados sugere que a redução de compostos sulfurados no diesel não induz suscetibilidade à contaminação microbiana, porém fatores como a origem da matéria prima, processo de refino e adição de biodiesel devem ser considerados. Os resultados desses estudos auxiliam na melhor compreensão dos microrganismos e metabólitos envolvidos nos processos de biocorrosão e biodeterioração do diesel e suas misturas visando a qualidade final do combustível. / Since 2008, the diesel fuel sold in the country has received the increasing addition of biodiesel, in addition to the gradual reduction in sulfur content, which is a global trend to minimize the impact generated by burning fossil fuels. Aspects related to the final quality of diesel and biodiesel blends, from production to storage, has received special attention. The aim of this study was to evaluate the susceptibility to microbial contamination during simulated storage, in pure diesel and blend B10 with different sulfur contents: ULSD (S10), LSD (S50), HSD (S500) and UHSD (S1800). The results of this research showed that Pseudallescheria boydii had greater ability to grow in the blend B10 than Paecilomyces variotii. The degradation analysis, infrared spectroscopy, showed that these fuels have different molecular characteristics according to their sulfur content. The data H1 NMR showed that the diesel with higher sulfur content (UHSD) showed a low level of branching and aromatic compounds compared to diesel fuel with lower sulfur content (ULSD). The high throughput sequencing (Illumina) of 16S and 18S rDNA showed the Phyla most abundant present in the mixed inoculum over time were: Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Ascomycota and Basidiomycota. Putative metabolites identified by UPLC / Q-ToF-MS in the water phase were: myristic, palmitic, stearic, oleic, linoleic and α-linolenic acids with relative abundance of 2-10 times higher than controls indicating degradation of microbial origin. Among the 10 evaluated parameters for chemical stability, only oxidative stability showed nonconformity with the ANP Technical Regulation and microbial contamination levels at the end of 90 days were considered acceptable. The data set suggest that the reduction of sulfur compounds in diesel fuel are not induced microbial susceptibility contamination, but factors such as the origin of the raw material, refining process and adding biodiesel to be considered. The results of these studies help to better understand the microorganisms and metabolites involved in the biocorrosion and biodeterioration processes of diesel and their blends aiming at the final quality of the fuel.