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CHANGES OF BACTERIAL SPECIES AND HEME PROTEIN OCCURRENCE IN ACTIVATED SLUDGE COMMUNITIES CULTURED IN THE LABORATORY
Title
CHANGES OF BACTERIAL SPECIES AND HEME PROTEIN OCCURRENCE IN ACTIVATED SLUDGE COMMUNITIES CULTURED IN THE LABORATORY
Creator
Wang, Xiaomeng
Date
2016, 2016-05
Description
An activated sludge sample that had originally been collected from an aeration tank of the Stickney wastewater treatment plant in Chicago, and...
Show moreAn activated sludge sample that had originally been collected from an aeration tank of the Stickney wastewater treatment plant in Chicago, and had previously been cultured at low dissolved oxygen (DO) for 48 weekly passages was used as starting material for continuation of the low DO acclimation. The culture was continued at low dissolved oxygen in synthetic wastewater for 25 additional weekly passages to study what would happen to the activated sludge if the low DO continued. In order to do that, some important data were measured during the culture, including the specific oxygen uptake rates (SOUR) which could reflect the ability of oxygen utilization, 16S rDNA information which could tell the community diversity of sludge, and the dominant species genome data which suggested what really happened to the sludge and some reasons. The results showed that SOUR decreased modestly during the course of low DO adaptation, which was contrary to the results of the previous study. There were significant changes in community structure with respect to bacterial species during the first fifteen additional passages. Species known to produce both flavohemoglobins (FHbs) and truncated hemoglobins (trHbs) were common at all passages tested, although the dominant species were totally different from passage to passage. Specifically, during the course of the experiment, the frequency of cells encoding an FHb decreased substantially, from 84% to 50%, while the percentage of cells encoding a trHb decreased slightly from 84% to 78%. The overall content in the culture of heme b (the heme type found in bacterial hemoglobins) decreased, however, during continuation of the low DO conditions. So it is indicated that the oxygen utilization ability of the activated sludge does not increase all the time.
M.S. in Biology, May 2016
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CHANGES OF BACTERIAL SPECIES AND HEME PROTEIN LEVEL IN ACTIVATED SLUDGE COMMUNITIES ACCLIMATED TO LOW AERATION
Title
CHANGES OF BACTERIAL SPECIES AND HEME PROTEIN LEVEL IN ACTIVATED SLUDGE COMMUNITIES ACCLIMATED TO LOW AERATION
Creator
Li, Hainan
Date
2016, 2016-05
Description
Increasing oxygen utilization of activated sludge under low dissolved oxygen (DO) conditions can lead to energy savings in wastewater...
Show moreIncreasing oxygen utilization of activated sludge under low dissolved oxygen (DO) conditions can lead to energy savings in wastewater treatment systems. Adaptation of sludge communities to low oxygen conditions may be facilitated by increased expression of heme proteins. One way to assess heme protein expression in sludge is to analyze the species present in the sludge community during the adaptation process. In the work reported here, growth of a sludge culture adapted by Kunkel to low aeration for 48 weekly passages was continued for an additional 26 passages (182 days). The activated sludge was cultured in synthetic wastewater under low DO conditions. Temperature, dissolved oxygen, and specific oxygen uptake rates (SOUR) were measured at the end of each weekly passage. The community structures of passages 50, 53 and 65 were determined by 16S rDNA cloning techniques. The type of heme present was identified using the pyridine hemochromogen method. The SOUR values measured increased slightly in the first part of the experiment, when the culture DO was very low, but then decreased in the latter part of the experiment, when the culture DO increased. The community structure diversity in passage 50 in Kunkel’s study and passage 50 from this work are totally different in terms of species present, even though the two cultures were both derived from Kunkel’s passage 48 and were grown under conditions which were matched as closely as possible for two additional passages. This indicates that the community structure is highly sensitive to small changes in growth conditions. From the point of view of types of hemoglobin (Hb), the community became more diverse by passage 53, containing all three types of truncated Hb (trHbN, trHbO, and trHbP), possibly due to the need for NO scavenging and oxygen transfer enhancement. The proportion of cells that synthesize truncated Hb decreased slightly through passages 50 to 53 to 65. The percentage of cells that encode FlavoHbs decreased from 84 % to 50 % by passage 65. Overall there was an increase in Hb expression from passage 50 to passage 65. Heme b expression in the sludge culture was confirmed. Successful adaptation of the sludge culture to low DO conditions via enhancement of oxygen uptake was not obvious in this study. Yet the overall tendencies of SOUR and Hb expression suggest that it is possible to acquire a culture more efficient in oxygen uptake if a stable low aerobic environment can be maintained for an extended period.
M.S. in Biology, May 2016
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