from collapsing homes. The incidence of cardiovascular disease (hypertension, 3 acute myocardial infarction, 4 heart failure, 4 pulmonary thromboembolism [PTE] 5,6 and deep vein thrombosis [DVT], 5 and other cardiovascular diseases 3,7-9), stroke, and respiratory infections 10 increases in evacuees after disasters. These concepts have been comprehensively reviewed. 11 The unsanitary environment of evacuation centers, sympathetic K umamoto Prefecture in Japan was struck by a series of earthquakes (2016 Kumamoto earthquakes 1,2), including a magnitude (Mw) 6.5 on 14 April and Mw 7.3 on 16 April 2016. After these earthquakes, >180,000 people were evacuated to evacuation centers or private vehicles. More than 3,000 aftershocks prevented vast numbers of evacuees from returning home and forced many to stay for a prolonged period in cramped conditions to escape

To isolate a biocatalytic enzyme, metagenomic libraries were constructed in fosmids from samples of activated sludge used to treat coke plant wastewater. Six indigo-producing clones were isolated from approximately 40,000 metagenomic clones in the search for the oxygenase responsible. In vitro mutagenesis and whole-sequencing revealed one open reading frame to be responsible for the production of indigo in the fosmid clones. The deduced sequence of the gene product showed 60% identity with 2-naphthoate monooxygenase from Burkholderia sp. JT1500. Subclones carrying this open reading frame (icpA) retained indigo production, and indigo-producing enzymes expressed from subclones catalyzed the oxidization of 4-nitrotoluene to form 4-nitrobenzyl alcohol. These results suggested that the icp product is an enzyme involved in catalyzing 4-nitrotoluene's oxygenation.

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