Eastman Chemical Company

Christine Jo Kelly

Gary S Sayler

Alice C Layton

Curtis A Lajoie

Raymond Stapleton

The invention provides two new zoogloeal strains, mz1t and mz2t. The invention provides an isolated nucleic acid consisting of the nucleic acid of SEQ ID NO:1. Examples of nucleic acids of mz1t include SEQ ID NO:2 and SEQ ID NO:3. An example of a nucleic acid of mz2t is SEQ ID NO:4. The invention also provides an isolated nucleic acid consisting of the nucleic acid of SEQ ID NO:5. A method of detecting the presence of zoogloeal clusters in a wastewater sample is provided, comprising: a) contacting RNA from a sample of the wastewater with a nucleic acid comprising the nucleic acid of SEQ ID Nos:1,2,3,4 or 5 under conditions that permit specific hybridization; and b) detecting the presence of hybridization, the presence of hybridization indicating the presence of zoogloeal clusters. A new Hyphomicrobium spp. strain, designated M3, is provided herein. The invention provides novel nucleic acids of a Hyphomicrobium sp. M3. For example SEQ ID NO:6 and SEQ ID NO:7 rDNAs of M3. SEQ ID Nos:8, 9, 10, 11, 12, 13 and 14 are presumptive Hyphomicrobium spp. sequences. SEQ ID NO:15 is a consensus sequence from 6m3 and 3m3 (Hyphomicrobium sp. m3 isolated from sludge. A method of detecting the presence of a Hyphomicrobium species in a wastewater sample as described above, using a nucleic acid comprising the nucleic acid of SEQ ID NO:17 is also provided. The nucleic acids of SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12 SEQ ID NO:13, SEQ ID NO:14 and SEQ ID NO:15 can also be utilized in whole or as fragments in the detection method for Hyphomicrobium spp.

Zoogloeal and hyphomicrobium spp. nucleic acids

The present invention provides a reporter bacterium, comprising a bacterium that occurs naturally in a biological sludge and that contains a nucleic acid that encodes a reporter protein not found in the naturally occurring bacterium. The nucleic acid can encode a bioluminescent reporter protein. A method and apparatus are also provided for detecting the presence of toxicity in a wastewater treatment influent stream, comprising contacting the influent with a reporter bacterium of the present invention; monitoring the expression of the reporter protein by the reporter bacterium; and correlating a reduction in the expression of the reporter protein with the presence of toxicity. In an additional embodiment, the present invention provides a method and apparatus for determining the effect of a known dose of an oxidative phosphorylation decoupling agent on biological wastewater sludge, comprising contacting a sample of the sludge containing a known dose of the decoupling agent and a control sample, with a reporter bacterium of the present invention; comparing the signal output by the reporter bacteria in the sludge sample containing the decoupling agent with the signal output by the reporter bacteria in the sludge sample not containing the decoupling agent; and correlating a decrease in signal output in the decoupled sample as compared to the signal output in the control sample with an action of the decoupling agent, whereby the effect of the known dose of the decoupling agent is determined.

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Philadelphia, TN, United States of America

Christine Jo Kelly