In aquatic environments the availability of nitrogen (N) generally limits primary production. N2-fixing prokaryotes (diazotrophs) can convert N2 gas into ammonium and provide significant input of N into the oceans. Cyanobacteria are thought to be the main N2-fixers but diazotrophs also include a wide range of heterotrophic bacteria. However, their activity and regulation in the water column is largely unknown. In this thesis the distribution, diversity, abundance, and activity of marine and estuarine heterotrophic diazotrophs was investigated. With molecular methods targeting the nifH gene, encoding the nitrogenase enzyme for N2 fixation, it was shown that diverse nifH genes affiliating with heterotrophic bacteria were ubiquitous in surface waters from ten marine locations world-wide and the estuarine Baltic Sea. Through enrichment cultures of Baltic Sea surface water in anaerobic N-free medium, heterotrophic N2 fixation was induced showing that there was a functional N2-fixing community present and isolates of heterotrophic diazotrophs were obtained. In Sargasso Sea surface waters, transcripts of nifH related to heterotrophic bacteria were detected indicating heterotrophic N2-fixing activity. Nitrogenase expression is thought to be highly regulated by the availability of inorganic N and the presence of oxygen. Low oxygen zones within the water column can be found in association with plankton. The presence of diazotrophs as symbionts of heterotrophic dinoflagellates was investigated and nifH genes related to heterotrophic diazotrophs rather than the cyanobacterial symbionts were found, suggesting that a symbiotic co-existence prevailed. Oxic-anoxic interfaces could also be potential sites for heterotrophic N2 fixation. The Baltic Sea contains large areas of anoxic bottom water. At the chemocline and in anoxic deep water heterotrophic diazotrophs were diverse, abundant and active. These findings extend the currently known regime of N2 fixation to also include ammonium-rich anaerobic waters. The results of this thesis suggest that heterotrophic diazotrophs are diverse and widely distributed in marine and estuarine waters and that they can also be active. However, limits in the knowledge on their physiology and factors which regulate their N2 fixation activity currently prevent an evaluation of their importance in the global marine N budget.

Submitted by Nádja Goes (nmoraissoares@gmail.com) on 2016-07-18T15:32:09Z No. of bitstreams: 1 2013_dis_loleite.pdf: 1004003 bytes, checksum: c14f43a7d45b40582533ad69e13cca53 (MD5) / Approved for entry into archive by Nádja Goes (nmoraissoares@gmail.com) on 2016-07-18T15:32:21Z (GMT) No. of bitstreams: 1 2013_dis_loleite.pdf: 1004003 bytes, checksum: c14f43a7d45b40582533ad69e13cca53 (MD5) / Made available in DSpace on 2016-07-18T15:32:21Z (GMT). No. of bitstreams: 1 2013_dis_loleite.pdf: 1004003 bytes, checksum: c14f43a7d45b40582533ad69e13cca53 (MD5) Previous issue date: 2013 / LEITE, Lana Oliveira. Avaliação do comportamento bacteriano e alterações sensoriais em tilápias do Nilo (Oreochromis niloticus) durante estocagem em gelo. 2013. 59 f. : Dissertação (mestrado) - Universidade Federal do Ceará, Centro de Ciências Agrárias, Departamento de Engenharia de Pesca, Fortaleza-CE, 2013 / Fish is a food susceptible to deterioration due their intrinsic characteristics, requiring bacterial monitoring in postmortem storage. Thus, the objective was to quantify mesophilic, psychrotrophic, psychrophilic bacteria in Nile tilapia (Oreochromis niloticus) during to 21 days of storage under the action of ice at temperature refrigeration. Four experiments were conducted, separated to two groups, gutted and ungutted, with eight samples totaling 32 samples. Every three days was taken a exemplar of tilapia for analyzes of the trial: sensory evaluation according to the Quality Index Method (QIM) adapted to the Nile tilapia and heterotrophic bacterial count of cultivable. In sensory analysis of raw fish attributes were assessed: general appearance, eyes, gills, muscles and texture of the fish, according to the Sensory Characters Sum (S.C.S). To each specimen were quantified bacterial groups: psychrophilic (23 °C), psychrotrophic (7 °C) and mesophilic (35 °C), the surface and the muscle, using the method of standard plate count (SPC). There was no statistical difference between the three bacterial groups analyzed the significance level of 5% within the same treatments and between groups. The storage time was highly correlated with the counting of bacterial groups and increased QIM for both eviscerated fish, and for those not drawn. It was considered that the process of evisceration on the premises of the fish did not increase its useful life. It is suggested, therefore, greater attention in the process of gutting fish markets that sell live fish, because this process was not carried out effectively to security and recovery product. / O pescado é um alimento susceptível à deterioração devido às suas características intrínsecas, sendo necessário o monitoramento bacteriano no armazenamento pós-morte. Dessa forma, objetivou-se quantificar bactérias, psicrotróficas, psicrófilas e mesófilas em tilápias do Nilo (Oreochromis niloticus) durante 21 dias de estocagem sob ação do gelo. Foram realizados quatro experimentos, separados em dois grupos, não eviscerados e eviscerados, cada um com oito amostras perfazendo um total de 32 amostras. A cada três dias era retirado um exemplar de tilápia para as análises do experimento: sensorialmente, conforme a avaliação do Método do Índice de Qualidade (MIQ), adaptado à tilápia do Nilo e microbiologicamente. Na análise sensorial do peixe cru foram avaliados os atributos: aspecto geral, olhos, brânquias, textura e musculatura dos peixes, segundo os somatórios dos caracteres sensoriais (S.C.S). De cada exemplar foram quantificados os grupos bacterianos: psicrotróficos (7°C), psicrófilos (23°C), e mesófilos (35°C), na superfície e no músculo, pelo método de Contagem Padrão em Placas (CPP). Não houve diferença estatística entre os três grupos bacterianos analisados à um nível de significância de 5% dentro dos mesmos tratamentos e entre os grupos. O tempo de estocagem apresentou alta correlação com a contagem dos grupos bacterianos e o aumento do MIQ, tanto para os peixes eviscerados, quanto para os não eviscerados. Foi considerado que o processo de evisceração no local de venda dos peixes não aumentou sua vida útil. Sugere-se, portanto, uma maior atenção no processo de evisceração em peixarias que comercializam peixes vivos, pois esse processo não foi realizado de forma eficaz para segurança e valorização do produto.

Deammonification is a widely used technology for side stream treatment with rich ammonium streams at relatively high temperatures, such as, the reject water coming from dewatering units in treatment of digested sludge and industrial wastewaters. The deammonification process has lower operational costs than conventional systems, consumes less energy, enables the increase of biogas production and it is easy to implement. However, this technology has not yet been applied in full- scale mainstream treatment due to its restrictions in coping with high C/N ratios, low temperatures, and the need for post-treatment processes. These conditions are allegedly negative to the growth and performance of anammox bacteria affecting the bacterial groups’ behavior in the process. This master thesis project aimed to evaluate the feasibility of using deammonification to remove nitrogen from mainstream wastewater, which was studied by monitoring the bacterial activity in a pilot scale reactor. The different bacterial groups involved (AOB, NOB, heterotrophs, and denitrifiers) were monitored by weekly measuring their activity in batch activity tests. The results allowed the evaluation of different operational scenarios and their impact by following up on the changes in the bacterial competition. The study was conducted for six months in a single-stage IFAS (integrated fixed-film activated sludge) pilot-scale reactor located in Stockholm and fed with pretreated (with a UASB) municipal wastewater. The different operational scenarios involved changes in temperature, aeration patterns, DO concentration, SRT, and HRT. The adjustment of these features was done in the interest of promoting AOB and anammox bacterial growth, leading to an improvement of the deammonification efficiency in future studies. However, the chosen operational conditions were to enhance bacterial competition and facilitate its visualization, not to maximize nitrogen removal. Thus, the most suitable scenario found during this study included DO concentration of 1.5 mg/L with 10 aeration-20 non-aeration pattern and ensured nitrogen removal rates within normal values while allowing the monitoring of all the bacterial groups. TN removal reached a value above 50% and NH4-N above 95%, whereas nitrogen Removal Rate (NRR) increased to 30g/N/m3-d and the system had an overall nitrogen removal efficiency of 75%. Nevertheless, it was proven that in the right environment, the necessary bacterial groups can be selectively accumulated and successfully perform deammonification and reduce nitrogen levels in mainstream wastewater. / Deammonifikation är en välanvänd teknik för rening av sidoströmmar med höga ammoniumkoncentrationer vid relativt hög temperatur, som till exempel rejektvatten från avvattning av rötslam eller industriellt avloppsvatten. Deammonifikationsprocessen har lägre driftkostnad än konventionella reningsprocesser, förbrukar mindre energi samt möjliggör högre biogasproduktion samtidigt som processen är enkel att implementera. Reningstekniken har dock ännu inte tillämpats i fullskala för rening av huvudströmmen på grund av den höga C/N-kvoten och de låga vattentemperaturerna i kommunalt avloppsvatten samt behovet av efterbehandling. Detta anses ha en negativ inverkan på anammoxbakteriernas tillväxthastighet och funktion vilket påverkar bakteriegruppens beteende i processen. Syftet med detta examensarbete var att utvärdera om det är praktiskt genomförbart att använda deammonifikation för att rena kväve från kommunalt avloppsvatten, vilket följdes upp genom att studera bakterieaktiviteten i en pilotskalereaktor. De involverade bakteriegrupperna (AOB, NOB, heterotrofer och denitrifierare) övervakades genom att mäta den mikrobiella aktiviteten varje vecka med hjälp av batch-tester. Resultaten användes till att utvärdera olika driftstrategier och deras effekt genom att följa förändringarna i mikrobiell aktivitet hos de konkurrerande bakteriegrupperna. Studien genomfördes i Stockholm under sex månader i en enstegs-IFAS-pilotskalereaktor (integrerad process med biofilm på fast bärarmaterial och aktivslam) som matades med kommunalt avloppsvatten som förbehandlats i en UASB-reaktor. De olika driftstrategierna omfattade olika temperaturer, luftningsstrategier, syrekoncentrationer, slamåldrar och hydrauliska uppehållstider. Syftet med driftstrategierna var att främja AOB- och anammoxbakteriers tillväxt för att i framtida studier kunna erhålla en förbättrad deammonifikationsprocess. Syftet i denna studie var dock i första hand att förbättra den bakteriella konkurrensen och göra den lättare att mäta, inte att uppnå bästa möjliga kväverening. Den driftstrategi som gav bäst resultat i denna studie innebar att hålla en syrehalt på 1,5 mg/l med 10 minuter luftning följt av 20 minuter utan luftning vilket säkerställde en normal kväveavskiljning och samtidigt möjliggjorde övervakning av samtliga fyra bakteriegrupper. Totalkväveavskiljningen var över 50 % och ammoniumavskiljningen över 95 % medan kvävereningsaktiviteten ökade till 30 g N/m3-d och systemet hade en övergripande effektivitet på 75 %. Studien visade att under rätt förutsättningar kan de nödvändiga bakteriegrupperna selekteras fram och deammonifikation av kommunalt avloppsvatten kan utföras på ett framgångsrikt sätt.

With a volcanic origin, Lake Kivu is deep and meromictic, and shows a very particular limnology and some astonishing features. The data available on its limnology and phytoplanktic communities are limited, dispersed or outdated. This is the first deep, long term study (2002-2004) on limnology and phytoplankton ecology of Lake Kivu, combining different techniques: HPLC analysis of marker pigments, flow cytometry, light, epifluorescence and electron microscopy. Lake Kivu combines a relatively shallow euphotic layer (~18m) usually smaller than its mixed layer (20 – 60 m), and with a weak thermal gradient in the mixolimnion. With an annual average chlorophyll a in the mixed layer of 2.2 mg m-3 and low nutrient levels in the euphotic zone, the lake is clearly oligotrophic. Concerning its phytoplanktonic composition, the most common species were the pennate diatoms Nitzschia bacata Hust. and Fragilaria danica (Kütz.) Lange Bert., and the cyanobacteria Planktolyngbya limnetica (Lemm.) Komárková-Legnerová and Cronberg and Synechococcus sp. Diatoms were the dominant group in the lake, particularly during the dry season episodes of deep mixing. During the rainy season, the stratified water column, with high light and lower nutrient availability, favoured dominance of cyanobacteria. Phycoerythrin-rich phototrophic picoplankton and heterotrophic bacteria cell numbers were constantly high, with relatively subtle spatial, seasonal and vertical variations. In open lake waters, where allochthonous carbon inputs are most probably inconsequential, HNA heterotrophic bacteria abundance is strongly correlated with chlorophyll a. Recent investigations revealed an increasing methane production in the deep waters during the past three decades, leading to an accumulation of gas and the subsequent lowering of the energy required to trigger a devastating release of gasses. The role of primary producers and the probable changes on the export ratio of the organic matter into deep waters after the introduction of the planktivorous sardine from Lake Tanganyika Limnothrissa miodon, is discussed. The actual primary production, 0.71 g C m-2 d-1 (~ 260 g C m-2 y-1), doesn’t seem substantially different from some punctual measurements made in the past, which discards the hypothesis of anthropogenic eutrophication. We believe that the export ratio of organic matter into the deep waters was largely affected by biological changes that could explain part of the methane increase in the past 30 years in Lake Kivu.

Diversity of the soil microbial community and its functional aspects were investigated in man-influenced environments, such as colliery spoil heaps in post mining sites and upland pasture used for outdoor cattle husbandry. The study was based on the cultivation of bacteria and streptomycetes as well as culture-independent approaches. Cultivated bacteria and streptomycetes were characterized by phenotypic and genotypic means. The culture-independent approaches were based on an analysis of environmental DNA in terms of both qualitative and quantitative parameters.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Atualmente, uma das maiores preocupações da humanidade é com a disponibilidade e qualidade da água. A exploração inadequada das fontes acarreta a contaminação das águas superficiais e subterrâneas que se tornam um risco para a saúde pública. Dentre as fontes biológicas de contaminação da água, incluem-se as bactérias, que são responsáveis por inúmeras doenças, sendo frequentemente tratadas com antimicrobianos. O amplo uso de antimicrobianos na medicina humana e na produção animal tem resultado no aumento do número de bactérias comensais e patogênicas resistentes a agentes antimicrobianos, fato que nos últimos anos vem se tornando um dos principais problemas de saúde pública. O presente trabalho resulta do interesse pelo estudo da qualidade da água consumida pelos alunos de 20 escolas estaduais da rede pública de Boa Vista RR, bem como obter o perfil de resistência a antimicrobianos de bactérias isoladas a partir de diferentes amostras de águas, coletadas nessas escolas. No período de outubro de 2010 a abril de 2011, foram coletas 40 amostras de águas de torneiras das copas e 40 amostras de águas de bebedouros, as quais foram submetidas à avaliação microbiológica. A determinação dos coliformes totais e termotolerantes foi realizada por meio da técnica dos tubos múltiplos com 3 diluições e série de 5 tubos. Para a detecção de bactérias heterotróficas uso-se o método pour plate, de acordo com as normas preconizadas no Standard Methods for the Examination of Water and Wastwater. O isolamento e a identificação dos isolados bacteriano foi realizada no LACEN-RR utilizando-se a série bioquímica. Para a avaliação da resistência aos antimicrobianos, utilizou-se a metodologia de difusão de disco, seguindo as normas estabelecidas pelo CLSI. Das amostras de águas que recebem tratamento completo, 10% apresentaram resultados positivos para coliformes totais e nas amostras de água clorada, 20% apresentaram resultados positivos para coliformes totais e 2,5% resultado positivo para coliformes termotolerantes, o que as tornam imprópria para o consumo humano. A contagem de bactérias heterotróficas variou de 1,4 x 10 a 3,6 x 102 UFC/mL de água, não ultrapassando o estabelecido pela Portaria 518/2004/MS. Foram identificadas as seguintes bactérias Gram-negativas (66,7%): Acinetobacter spp., C. violacea, Cocobacilos spp., E. aerogenes, E. coli, K. ozaenae, Leptothrix ssp. e Bacilo Gram-negativo não fermentador de glicose, as bactérias Gram-positivas (33,3%), identificadas foram: B. subtilis, Micrococcus spp., S. aureus e Staphylococcus spp. Os resultados dos testes de sensibilidade aos antibióticos revelaram que a maioria das bactérias isoladas foi resistente a ampicilina (68,2%), 27,3% apresentaram resistência a oxacilina e 22,7% foram resistentes a eritromicina e meropenem. As cepas de S. aureus isoladas das amostras de águas apresentaram maior índice MAR (0,61) entre as bactérias Gram-positivas, enquanto que nas bactérias Gram-negativas, as cepas que apresentaram maior índice MAR foram as cepas de E. coli, com índice 0,61. Os resultados comprovam a ocorrência de instabilidade no padrão de qualidade da água fornecida a população de Boa Vista RR seja no tratamento, na distribuição ou no armazenamento da água. Além de constatar a presença de bactérias multirresistentes a antibióticos, o que representa risco à população. / One of the main concerns of humanity nowadays is water: its availability and its quality. An inadequate exploitation of the sources results in the contamination of both superficial and deep waters, which become a public health hazard. Bacteria are one of these biological sources of water contamination, responsible for countless diseases which are treated with antimicrobials. The widely spread use of antimicrobials in human medicine along with animal production had resulted in a steady increase in the number of pathogenic and commensal bacteria resistant to antimicrobial agents, which in recent years are becoming one of the main problems of public health. This paper is a result of an interest in studying the quality of water consumed by the students of 20 state schools of the public school system of Boa Vista RR, as well as trying to obtain a profile of antimicrobial resistance of the bacteria isolated in different water samples collected at these schools. Between October 2010 and April 2011, 40 water samples were collected from taps in lunchrooms, along with 40 samples from drinking wells, and all of them were submitted to microbiological examination. Using the multiple tubes technique, with 3 dilutions and a series of 5 tubes, we determined the total number of coliforms and thermotolerant coliforms. In agreement with the rules of the Standard Methods for the Examination of Water and Wastewater, the pour plate method was used at the CBio UFRR to detect heterotrophic bacteria. Isolation and identification of microorganisms was done at LACEN RR, using the biochemical series. In evaluating the resistance to antimicrobials, disk diffusion methods were used, in agreement with the established rules of the CLSI. In water samples that receive full treatment, 10% tested positive for total coliforms and, in chlorinated water samples, 20% tested positive for total coliforms and 2,5% for thermo-tolerant coliforms, which makes them inadequate for human consume. The counting of heterotrophic bacteria went from 1,4 x 10 to 3,6 x 10 CFU/mL of water, not surpassing what is established in the ordinance 518/2004/MS. The following Gram-negative bacteria (66,7%) were identified: Acinetobacter spp., C. violacea, Cocobacilos spp., E. aerogenes, E. coli, K. ozaenae, Leptothrix ssp. and Gram-negative glucose non fermenting bacillus. The Gram-positive bacteria identified were: B. subtilis, Micrococcus spp., S. aureus and Staphylococcus spp. The results of antibiotic sensibility tests revealed that most of the isolated bacteria were resistant to ampicilin (68,2%), 22,3% showed resistance to oxacilin and 22,7% were resistant to both eritromicin and meropenem. In Gram-positive bacteria, the growths of S. aureus isolated from water samples showed the greatest MAR index (0,61), while E. coli growths were the ones who showed the greatest MAR index in Gram-negative bacteria (also 6,1%). Results prove a great instability in the quality of water provided to the Boa Vista - RR population, being in its treatment, distribution or containment, as well as showing the presence of bacteria resistant to multiple antibiotics, which is a health hazard to the public.