A PROPOSAL
The Anacostia Veterans Project
Presented by: The National Association of Concerned Veterans (a 501c3 non-profit Corporation)
CONTENTS
Section One
Nano Technology 3
A Green Technology Business 4
Municipal Water Supply 5
Primary Markets of Opportunity 6
Secondary Markets 7
Section Two Phase One: Proof of Concept
Rationale 8
The National Association of Concerned Veterans (NACV) 9
The Anacostia Project Design Approach
Statement of Work 10
Management 11 Expenses 12
Appendices #1 Fantastic Properties of Nano-bubbles #2 WATER - Water Quality and Microbial Community Changes in an Urban River after Micro-Nano Bubble Technology in SituTreatment
Section Two
Phase One: Proof of Concept
Rationale
Background
The National Association of Concerned Veterans - At the end of the Second World War, America occu- pied a position of unprecedented opportunity and prosperity. Millions of men and women returned home. They came back to a nation transformed by the experience of victory - a nation catapulted into a new era of global influence and technological innovation. After the long night of austerity. rationing and shortages, a veritable explosion of demand, productivity, and economic growth was about to begin. Advances in engineering, science, & medicine opened doors to all. Education, thanks to the GI Bill, thrived and male life expectancy jumped 10+ years. There was a great challenge in the land. We had a new Nation to build. It was the ascendency of the American Century. Veterans played a lead role in that era of prosperity.
Then came Korea, Viet Nam, and a seemingly endless series of global engagements. America’s growth in the 50’s and 60’s became clouded by new responsibilities as we assumed the role of leader of the “free world.” The proxy wars in Africa and Anti Terrorism campaigns began to create a new gen- eration of veterans. A broad multi ethnic class of veterans began to emerge. The challenges and op- portunities for our veterans began to shrink at home as global economies shifted and changed abd as the sense of national mission shifted and changed. As a country we seemed to lose our focus on the sacrifice of service and while recently a new emphasis in both a national veterans awareness dialog and the resources devoted has improved, there exists a gap in the role and place of our bravest and most vulnerable population segment: America’s Military Veterans.
A New Vital Mission:
America’s Veterans by their enlistment, signed a blank check to the USA for any amount up to and in- cluding their very lives. Nothing so concentrates the mind as being the subject of ‘incoming fire,” That is as “real,” as life gets. Our veterans by, so doing became the defenders of our nation and our democ- racy. Now our country now faces another challenge. The world is soon to become affected by funda- mental changes in the planetary economy and the environment. NACV believes that our mission driven dedicated veterans, who so abely defended this nation from threats to our liberty - Now defend us against the equal or greater threat to our planet’s environment. Just as they defended and became guardians of our liberty, let us train them for a new and equally vital mission - The defense of our envi- ronment through Food and Water security. Let it start here in Washington, The District of Columbia. 8
The National Association of Concerned Veterans (NACV)
Organized in 1968 has a long history of training, hiring, and assisting veterans of America’s wars,. We started in a time and place where the Veteran was not being supported and often operated under less than ideal circumstances. However, like all of our nation’s veterans, and as vets ourselves, we perse- vered in the face of adversity. We are resourceful and dedicated to our Country and the City’s in which we reside. The following proposal sets forth a strategic approach to achieving that goal.
The Anacostia River Watershed has long been an embarrassment to our city and a blot on the National Capitol, The City of Washington, DC. While great progress has been made and recent technical reports are most encouraging, there are still water quality issues that need to be resolved. We believe we have the technology and expertise to complete the final stage of the full ecological and environmental restoration of the Anacostia Watershed.
The Anacostia was once a pristine river, it’s waters the natural habitat for a variety of native waterfowl, fish, and aquatic flora and fauna. A growing population, then the industrial revolution began to take its toll.and the river fell into a period of massive pollution and environmental ruin. In recent years the Federal Government and successive city administrations have led a concerted effort to bring this once bucolic river back to its vital andlife affirming health and beauty.
DC Veterans - The most recent Unemployed Veterans Study places The District of Columbia with the highest unemployment rate for Vets in the Nation. (2015) However, since then the Veterans Administra- tion the The DC Government have worked together to substantially reduce this number.and numerous support programs now exist. The NACV proposes to train and productivelly employ a new generation of concerned and engaged veterans to install, maintain and operate, a new, totally natural, chemically free means to bring the river back to its full capacity and to once again support life in all the natural eco- logical diversity
.NBO technology (...nano bubbles of oxygen) is a Japanese developed process used to revitalize and repair the littoral waters of Japan and it’s fishing grounds from a massive dump of organic matter dur- ing the last Tsunami. NB are a safe natural process of proven efficacy (see appendix “A”)
Complete Remediation - Anacostia River Watershed Bioremediation can be completed and stabilized using strategically placed and maintained Nanobubble Ox Generators. It is a cost effective holistic so- lution that can be totally managed by our Veterans who can be the protectors of our environment as they were the protectors of our liberty. Here is a link to a Japanese video of a similar project in Japan - https://binged.it/3424zic
9
The Anacostia Project Design Approach
Overall Strategy - It is not our intention to re invent or to duplicate existing engineering but to collabo- rate and augment positive existing efforts through the application of the unique properties and advan- tages of nanobubbles (see appendix #1)and to restore the original ecology of The Anacostia Water- shed. Our model is designed to help facilitate the clean up of the river and to then maintain Its ongoing health through the proactive super oxygenation of the water and the restoration of the natural flora and fauna native to this region.
Conceptual Design - NACV plans a design based upon both content and basic engineering principles:
1. Review literature, records, and work to date
2. Analyze all available data points
3. Create a project design
4. Implement a work plan
5. Developmentally test outcomes
6. Monitor, improve and maintain system
Statement of Work
Phase One: Program Objective:
Quantitatively establish the efficacy of nano technology (nanoponics) in restoration of the natural ecology of The Anacostia Watershed
NACV will perform the following work tasks associated with a professional engineering project. Listed below is a preliminary estimate based upon our expertise and the limited knowledge of the existing condition of the Anacostia and prevailing efforts to date.. Upon award and prior to the beginning of ef- fort, NACV will prepare a detailed statement of work that will enumerate and detail the exact number of personnel, type model, & cost of all equipment required, as well as the job tasks necessary to a suc- cessful project. The work plan will detail and identify the following:
Quantifiable objective of the project
1. Strategic engineering design
2. Timing of technical reviews
3. Specific tasks
4. Human resources and skills required
5. Training plan & syllabus
6. Project timeline
7. Deliverables (physical items)
Management
Overview - NCVA anticipates that engagement with the city will be through a cost plus contract. Over- head and profit will be subject to city regulations for such projects. NCVA will hold the master agree- ment and will manage and have primary responsibility to the City on all aspects, technical, personnel, and the role of subcontractors as required. CVA will provide a statement of financial responsibility to include a current copy of its 990 filing, as well as, a board resolution authorizing this project.
Project Management - NCVA establish a project overview team that will include NACV, City Project Manager, and a selected representative of all ancillary contractors and stakeholders. This Project Oversight Team will meet periodically to be briefed and review current plans and status and to provide insight and mutual reciprocal suggestions and improvements . .NACV will also employ a professionally certified Project Management Professional (PMP) who will have responsibility for both management and reporting on all aspects of the project to include: compliance, federal/city regulations, financial is- sues, personnel hiring etc, etc. as may be required by the City. Regular project reports and meetings will be held so that the city may exercise its right to oversee the efficacy of the project.
Hiring Practices - NACV will hire and train veterans in all aspects of the operation. Where no veteran with the skills can be identified, we will find a candidate we can train for the position while issuing a short term contract for a suitable subject expert. Veterans will be a first priority followed by: Returning Citizens, Single Parent (head of households) Endangered Youth, and the LGBT community will also be priority hires.
Terms & Conditions In collaboration and consultation with all engaged parties and stakeholders NACV will select a target section of the Anacostia as the test site. NCVA will form a technical advisory committee methods, we will deploy the NB infiltration process and conduct weekly assays of the key measures as determined by the joint committee. We anticipate a three month test period from inception to final proof of concept. Although formal testing will occur every 5 days and the results carefully recorded and monitored, infor- mal testing and operational validation will occur periodically as well. The official testing will begin once all equipment is received and installed per to NACV’s satisfaction.
Expenses
Relevant expenses (travel & per diem will be billed as standard GSA rates) and will be pre approved by client. Approved expenses will be considered extra and billed as incurred or billed against a city issued debit card.
Estimated Budget: It is anticipated that the city will, under the general terms and conditions of the McKinney Act. Identify surplus locations and such equipment and resources as may be available through either state or federal gov’t programs. This will mitigate project costs significantly. Salaries for professional staff will be consistent with market rates generally paid for such work. Veteran participants will be paid a living wage of $17.00 dollars per hour based upon standard federal and city personnel requirements. Wages paid will be determined at the time a formal contract is signed. The following are estimated for planning purposes only. Facility: 10,000 sq ft with space for office, classroom, workspace, conference + Leasehold Improve- ments. This includes office equipment: Standard Office resources
Estimated Staffing - Compensation: We estimate that initial level of effort will not require all staff to be full time employees.
● 1 Program Manager - full time @ $50K per annum
● 1 Engineer - part time @ $33.00 per hr.
● 1 Trainer part time @ $23.00 per hr.
● 2 Technicians - part time @ $20.00 per hr.
● 4 Staff: New Hires - full time @ $17.50 per hr.
Estimated Compensation* (Reflects 18% benefits etc, (required by all Gov’t contracts)
Estimated Equipment Costs - For Test Completion
No # Name & Type of Equipment Function Estimated Cost
2 Submersible Pumps Water $12,500
O2 injection $40,000
1 Nano Scale Precision Nozzle Creates NB mix $20,000
Controls & Automation Measurement Control. Programming
$$5,520 $4,800 $8,400
$39,220
$22,000
- Misc. tools, piping, plumbing Interconnection of $15,000 system
1 Bio Remediation Consultant Ecological restora-
tion expertise 12 $10,000
Total Estimated System Costs
$107,000
Staff Salaries: $47,500
Equipment Costs 107,000
Contingency @ 50% 53,500 Expert Consulting @ 50% 77,250
Sub Total 285,250 Profit @ 7% 19,968
Total Estimated Cost $305,218
Appendices
Water Quality and Microbial Community Changes in an Urban River after Micro-Nano Bubble Technology in Situ Treatment
Yifei Wu 1,2 , Hui Lin 3, Weizhao Yin 4, Sicheng Shao 1,2, Sihao Lv 3,* and Yongyou Hu 1,2,*
1 School of Environment and Energy, South China University of Technology, Ministry of Education, South China University of Technology, Guangzhou Higher Education Mega Centre, Guangzhou 510006, China; 201620132571@mail.scut.edu.cn (Y.W.); sicheng2009010370@163.com (S.S.) 2 The Key Lab of Pollution Control and Ecosystem Restoration in Industry Clusters, Ministry of Education,
South China University of Technology, Guangzhou Higher Education Mega Centre, Guangzhou 510006, China 3 Research Center for Eco-Environmental Engineering, Dongguan University of Technology,
Dongguan 523808, China; linhui@dgut.edu.cn 4 School of Environment, Jinan University, Guangzhou 510632, China; weizhaoyin@jnu.edu.cn * Correspondence: lvsh@dgut.edu.cn (S.L.); ppyyhu@scut.edu.cn (Y.H.)
Received: 6 December 2018; Accepted: 27 December 2018; Published: 2 January 2019
Abstract: Currently, black-odor river has received great attention in China. In this study, the micro-nano bubble technology (MBT) was used to mitigate the water pollution rapidly and continuously by increasing the concentration of dissolved oxygen (DO) in water. During treatment, the concentration of DO increased from 0.60 mg/L to over 5.00 mg/L, and the oxidation reduction potential (ORP) also changed from a negative value to over 100.00 mV after only five days aeration. High throughput pyrosequencing technology was employed to identify the microbial community structure. At genus level, the dominant bacteria were anaerobic and nutrient-loving microbes (e.g., Arcobacter sp., Azonexus sp., andCitrobacter sp.) before, and the relative abundances of aerobic and functional microbes (e.g., Perlucidibaca sp., Pseudarcicella sp., Rhodoluna sp., and Sediminibacterium sp.) were increased after treatment. Meanwhile, the water quality was significantly improved with about 50% removal ratios of chemical oxygen demand (CODCr) and ammonia nitrogen (NH4+-N). Canonical correspondence analysis (CCA) results showed that microbial community structure shaped by COD, DO, NH4+-N, and TP, CCA1 and CCA2 explained 41.94% and 24.56% of total variances, respectively. Overall, the MBT could improve the water quality of urban black-odor river by raising the DO and activate the aerobic microbes.
Keywords: long term performance; black-odor urban river; micro-nano bubble technology; in situ treatment; biodiversity
1. Introduction
Urban watercourses are essential and important to the urban ecosystem, landscape ecology and citizen’s life. However, in recent decades, rapid industrialization and urbanization has accelerated the worsening waters in urban area in China. Urban rivers are usually used as drains for sewage, industrial wastewater and even domestic garbage, which contained various pollutants. When organic pollutants are discharged to rivers, part of them is precipitated in the sediment and the others are metabolized by microbes [1,2]. During the metabolism, the microbes exhaust the dissolved oxygen in the water and make the aquatic environment anoxic, which promotes the growth of anaerobic microbes. Meanwhile, the anaerobic digestion produces odor gases (e.g., H2S) [3] and black substances
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water
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(e.g., FeS) [4,5]. As a result, the river is in anoxic condition, showing black appearance and stinking smell. These black-odor rivers flows through the cities affect residents’ life and pose a serious threat to human health [6]. Thus, treating the polluted urban rivers and recovering their ecological function have attracted much attention in China.
It is well known that microbes play an important role in biogeochemical cycling of aquatic ecosystem, which could decompose, transform and mineralize organic or inorganic matters in urban rivers [7]. Additionally, microbes are the first that interacted with dissolved substances and severely impacted by perturbations of the water quality [8,9]. Previous study showed that although microbes could adapt to the environment fluctuation to some extent, the microbial structure will shift significantly with intensive change of the ambient [10]. Thereby, microbes could be used to monitor and evaluate the pollution of water, and increasing the concentration of dissolved oxygen in the water body is the primary method to alleviate the pollution of urban streams.
Many physiochemical or biological treatment methods, which involve plants, aeration, chemical agents and bacterial inoculation exclusively or in combination to remove or metabolize the pollutants [11–13], and to mitigate the pollution in different extents. Lately, the micro-nano bubble technology (MBT) has been applied in water treatment, resulting in good effects. Micro-nano bubbles are small bubbles with diameters of several micrometers and nanometers, with high self-pressurization and longer lifetime [14,15], which have recently been explored as promising candidates for aeration. The primary application of micro-nano bubble was focused on activating the microorganisms, flotation, water treatment, and aeration [14,16,17]. Unlike the normal macro bubbles (with diameter 1–10 mm), the micro-nano bubbles would swell and burst in liquid with high mass transfer [15], and significantly increase the concentration of dissolved oxygen (DO) in water, reaching to the possible oversaturation. For these excellent properties, MBT would resolve the problem of hypoxia in polluted river efficiently. Recently, micro-nano bubbles were used to mitigate the pollution by enhancing the concentration of DO and eliminating the impurities in the river [18].
In this study, MBT was used for aeration and enhance the concentration of DO in an urban black-odor river, and the long term performance of MBT treatment was investigated. In the Pearl River Delta, featuring highly-developed manufacturing industry, a river was picked to be the trial target of MBT experiment. The object was figured out the MBT influence on microbial community structure, explored the effect on DO, oxidation reduction potential (ORP), chemical oxygen demand (CODCr), ammonia nitrogen (NH4+-N), pH, and total phosphorus (TP) [19]. The relationship between microbes and physicochemical factors of water was also investigated.
2. Material and Methods
2.1. Study Site and Sampling
The urban river lies between 22◦45 24” N, 113◦47 9” E and 22◦46 13” N, 113◦46 52” E, the city of Dongguan, Guangdong Province, South China. This area is dominated by a typical subtropical marine climate with annual average temperature around 22.2 ◦C, data are given in Figure S1 in the Supplementary Information (SI). This river is a tributary of Maozhou River, which is connected to the South China Sea.
Considering the past influences of the drain outlets along the rivers and tidal water, we set up the sampling sites at the upper reaches of the river. In this study, three sampling sites (marked as S1 (22◦46 8” N, 113◦46 51” E), S2 (22◦45 53” N, 113◦46 53” E), and S3 (22◦45 43” N, 113◦46 57” E)) were picked at the deep areas along the river, with an over 300 m distance from each other (Figure 1). To study the long-term performance of MBT, the sampling work lasted for 12 months from 19 July 2017 to 27 August 2018, including 3 periods. The first period was from day 0 to day 3, which showed the original physiochemical properties of the river before treatment. The second period was from day 4 to day 39, which was the continuous aeration period (aeration for 24 h per day). The third period was from day 40 to day 157, which was intermittent aeration (aeration for 6 h per day). The work was paused
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during February (28 days), 2018 for the Spring Festival. During the treatment, we sampled every day before day 40, and sampled every Monday, Thursday, and Saturday from day 40 to day 157. Samples for microbial community research were collected on 20 July (d1), 25 August (d20), 30 September (d37), 30 October (d67), 30 April (d133), and 31 May (d146), respectively. The temperature was around 25~35 ◦C when taken with the microbial samples to minimize the influence of temperature. Samples for CODCr, NH4+-N and TP analysis were collected with a van Dorn bottles under the surface of river (0.5 m) and sent to the laboratory immediately. The DO, pH, and ORP were measured in situ at the subsurface using a multipara meter quality detector (Hydrolabi DS5, HACH, USA). The concentration of TP, COD and NH4+-N were measured according to the standard methods (APHA, 2012) [20]. The ORP was only monitored from day 0 to day 39 for confirming its relationship with DO. Before the with treatment, DO. Before all the the drain treatment, outlets were all the blocked. drain outlets were blocked.
Figure Figure 1. 1. The The location location of of Dongguan Dongguan City City in in China China (A), (A), the the location location of of Changan Changan Town Town in in Dongguan Dongguan City (B), the location of the river in Changan Town (C), and the sampling sites of this study (D).
2.2. DNA Extraction and Illumina Miseq Sequencing
An E.Z.N.ATM Mag-Bind Soil DNA Kit (Omega, Bio-Tek, Norcross, GA, USA) was employed to extracted DNA by following the manufacturer’s instructions. The PCR amplification was performed according to the Illumina 16S Metagenomic Sequencing Library preparation guide (Illumina). The V3-V4 hypervariable regions of the bacteria 16S rRNA gene were amplified with primers 341F (5 -CCTACGGGNGGCWGCAG-3 ) and 805R (5 -GACTACHVGGGTATCTAATCC-3 ) by the thermocycler PCR system. The purified amplicons were measured by Qubit dsDNA HS Assay Kit (Life Technologies, Eugene, OR, USA). The purified amplicons were pooled in equimolar, using the Illumina MiSeq system by Sangon Biotech (Shanghai, China). Sequences containing ambiguous bases and any longer than 480 base pairs (bp) were dislodged and those with a maximum homopolymer length of 6 bp were allowed [21]. Any sequence shorter than 200bp were removed.
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2.3. Statistical and Bioinformatics Analysis
Raw fastq files were de-multiplexed and quality-filtered using QIIME (version 1.17). The OTUs were clustered with 97% similarity [22] by UPARSE (version 7.1) [23], and chimeric sequences were identified and removed using UCHIME [24]. In this study, MOTHUR [25] was used to calculate the Shannon’s diversity, Simpson index, ACE and Chao 1 for each sample. Coverage is defined as: C=1 − n1N , where n1 refers to the number of the singleton OTU, N is the number of the total sequences. ACE and Chao 1 can estimate the species richness.
The hierarchical clustering tree on OTU level was established based on Bray-Curtis distance with Vegan package within R. The principal coordinate analysis was based on the UniFrac dissimilarity values and performed to interpret the microbial communities’ relative similarity from each sample. The Canonical correspondence analysis (CCA) was used to determine the correlations between microbial communities and environmental factors by Vegan package within R. The microbial community functions were predicted by Clusters of Orthologous Group (COG).
3. Results and Discussion
3.1. Water Quality Improvement of Black-Odor River
Selected water physicochemical indexes, such as DO, ORP, COD and so on, were used to evaluate the river condition and impact assessment. The result showed that the river was polluted with low DO (0.60 mg/L–1.02 mg/L) and ORP (around −140.00 mV) levels before treatment, and then the DO and ORP values were significantly increased during MBT treatment and stabilizes gradually (Figure 2). During the continuous aeration period, the DO increased to 4.00 mg/L–7.50 mg/L, and then were stable around 4.00 mg/L during intermittent aeration (Figure 2A). In like manner, the ORP value increased from negative values to around 130.00 mV during continuous aeration period, and then stable. The result in March 1 showed that the DO was decreased after one month without aeration due to the Spring Festival, while it was recovered to previous values after several days of intermittent aeration. The sharp decreasing of DO and ORP values at day 20 might due to the discharge of waste-water when sampling. Meanwhile, the pH was around 7.00 during the treatment, which was not affected by MBT. In addition, the micro-nano bubbles could combine the impurities in the river, then float upon water and been eliminated, for which, the turbidity of the river was decreased from 148.00 NTU (d1) to around 21.00 NTU (d157) (data not shown). decreased from 148.00 NTU (d1) to around 21.00 NTU (d157) (data not shown).
Figure 2. Cont.
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Figure Figure 2. 2. The The DO DO and and ORP ORP improvement improvement of of river river (A: (A: DO, DO, B: B: ORP). ORP).
A massive amount of nitrogen and phosphorous in aquatic environment led to eutrophication of urban rivers. In this study, the concentration of NH4+-N was increased at the beginning of aeration and then decreased to a lower value (Figure 3A). The change from continuous aeration to intermittent aeration had affected the concentration of NH4+-N, which was steady gradually at the following time. The concentration of NH4+-N was increased during the cold days, which might relate to the low activity of microbes with low temperatures [26]. Site 3 was closed to the Maozhou River, and diluted by the tidal river, resulting in lower concentration of NH4+-N after aeration. During the sampling period, the concentration of TP didn’t decreased as other factors. The aeration even promoted the phosphorus release from sediment and caused the values a little higher than before, data are given in Figure S2 in SI. Meanwhile, the CODCr value could reflect the extent of river pollution and pollution level. After treatment, the CODCr of the river decreased significantly (Figure 3B) and stayed at a low level around 45.00 mg/L. The CODCr value increased after February and decreased to former concentration in a few days with intermittent aeration. The results showed that the intermittent aeration is necessary for guaranteeing Water 2018, 10, x the FOR water PEER REVIEW quality. 6 of 14
Figure 3. Cont.
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Figure Figure 3. 3. The The NHNH 4+-N -N and and COD COD improvement improvement of of river river (A: (A: NHNH 4+-N, -N, B: B: COD). COD).
At the beginning of aeration, the value of DO and ORP was significantly increased, meanwhile the aeration also promoted the organic matters releasing from sediment, therefore, the concentration of organic matters was increased first and was then followed by a decreasing trend. Finally, the physicochemical properties of river were improved and stabilized gradually at the later stage of treatment. It should be noted that the long-term intermittent aeration would be necessary to ensure the water quality of the polluted river if the internal pollution was not done.
3.2. Change of Microbial Profile Alongside MBT Treatment
The microbes would respond to the change of aquatic environment, such as the physicochemical properties, organic and inorganic matters. The number of clean sequencing was changed from 59,485 to 32,237, and the coverage was more than 98.50%, which presented the real microbial information in the river. Before the treatment, there were 1845 of OTUs in the river (day 1); at day 37, there were 1603 of OTUs; at day 146, there were 968 of OTUs. The results showed that the OTU number was changed with the water quality of the river, which showed a decreasing trend. The Chao 1 and ACE indices showed that the microbial community richness was decreased after MBT treatment, data are given in Table S1 in SI. However, the Shannon and Simpson indices had little change, indicating that the microbial evenness was higher after treatment. There were 224 to 565 unique OTUs were detected during the treatment, indicating that the MBT treatment activated some indigenous microorganism at different stage of the treatment. The decreasing trend might be related to the decreasing concentration of organic matters, the lower nutrition for microbial growth in the river, the lower microbes’ richness and diversity. At the same time, the microbes, which adapted to the condition of river, would become the new dominant species.
To illustrate the relationship among microbial communities from different samples, the cluster tree using Bray-Crutis were constructed according to the OTU distribution at 97.00% similarity level. According to Figure 4, the six samples were clustered into four major groups: The first group was composed of d1 and d20, d67, and d133 was the second and third group separately, d37 and d146 were the fourth groups. According to the previous study, the microbial community would shift with the ambient [27], which demonstrated that the river conditions were similar in every single group. In total, the results showed that microbial community would shift with the changing environment in urban river during the treatment by MBT, and the biodiversity showed a decreasing trend with the improvement of river quality.
Water improvement 2019, 11, 66 of river quality.
7 of 14 Figure Figure 4. 4. Bray-Curtis Bray-Curtis tree tree based based on on the the 97% 97% similarity similarity of of the the OTUs.
OTUs.
3.3. Change of the Microbial Community Structure
The change of microbial community composition is the key point of microbial impact of MBT. For better understanding the succession of microbial community during the treatment, the taxonomic affiliation at class level and genus level were analyzed.
3.3.1. The Microbial Community Succession at Class Level
The Figure 5A shows the distribution of microbes at class level (abundance ≥1%). The Alphaproteobacteria, Betaproteobacteria, Deltaproteobacteria, Epsilonproteobacteria, Gammaproteobacteria, Flavobacteriia, Actinobacteria, Clostridia, Sphingobacteriia, and Bacteroidia were dominant microbes at class level. The Alphaproteobacteria, Gammaproteobacteria and Deltaproteobacteria are usually abundant in marine environment [28]. The tidal water brought seawater and marine microbes, affecting the microbial community structure in the river. For other microbes, though they were still abundant after treatment, the relative abundance was changed. The relative abundance of Actinobacteria, Cytophagia and Sphingobacteriia were increased significantly after MBT treatment, which were found in oligotrophic aquatic environment and able to degrade biopolymer [29]. Furthermore, it was reported that Actinobacteria and Verrucomicrobiae are more abundant in sediment samples than water [30]. The increased abundance of Actinobacteria and Verrucomicrobiae indicated that MBT might encourage the microorganisms migrated with nutrients from sediment to water.
The Bacteroidia, Clostridia, Deltaproteobacteria, Flavobacteriia and Synergistia were decreased after MBT treatment, most of which were reported as potential opportunistic pathogens or nutrient-loving microbes under anaerobic condition. For example, Bacteroidia and Flavobacteriia were reported as potential fecal indicator bacteria [31], Deltaproteobacteria could metabolize sulfate in anaerobic environment [30]. The decreased trend of these microbes suggests that after MBT treatment, the aquatic environment was not suitable for these microbes. In all, the result showed that MBT would influence the abundance of microbes, which was mainly related to the concentration of DO and organic matters.
Water 2019, 11, 66 8 of 14 organic matters.
A
Figure 5. Cont.
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OR PEER REVIEW
B
Figure 5. Community structure and distribution of samples (A: At class level by Circos analysis; B: At genus level by heatmap).
3.3.2. The Microbial Community Succession at Genus Level
The Figure 5B showed the response of predominant genera (abundance ≥ 0.1%) to MBT treatment, Acidovorax, Acinetobacter, Cloacibacterium, and Novosphingobium were abundant before and after MBT treatment, living in fresh water commonly [32]. The abundance of Arcobacter sp., Azonexus sp., Citrobacter sp., Dechloromonas, Flavobacterium, Rhodobacter sp., and Geobacter sp. were decreased after treatment, which might be related to the increasing DO and the decreasing concentration of organic matters [33,34]. In contrast, the abundance of Perlucidibaca sp., Pseudarcicella sp., Rhodoluna sp., and Sediminibacterium sp., were higher after MBT treatment. Most of these genera were aerobic bacteria and functional microbes, such as Sediminibacterium sp. and Rheinheimera, which were aerobic microbes and predominant in freshwater, marine and sediment [35,36].
At different stages of treatment, the dominant microbes were different. For example, before treatment, the dominant microbes prefer anaerobic and eutrophic condition (e.g., Dechloromonas and Flavobacterium); during the treatment, the abundance of sediment microbes were increased
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(e.g., Actinobacteria and Verrucomicrobiae); and after treatment, the abundant microbes were adapted to aerobic and oligotrophic environment (e.g., Actinobacteria, Cytophagia and Sphingobacteriia). It is clear that the microbial community would shift with the concentration of pollutants and changing environment [37].
3.4. Correlation of Microbial Community and Physiochemical Characteristics
The Canonical correspondence analysis (CCA) was used to determine the relationship between microbial community and physiochemical properties in the river, and five variables (CODCr, NH4+-N, pH, DO and TP) were selected to form a significant CCA model (p < 0.05). CCA1 and CCA2 presented 41.94% and 24.56% of total variances, respectively (Figure 6), indicating that these five variables were the major factors shaping microbial community.
In Figure 6, DO, COD and pH were the major factors shaping the microbial community structure, NH4+-N and TP were also involved. The previous study showed the importance of DO for microbial activity in aquatic environment [38], and the concentration of COD and NH4+-N would also influence the microbes [39]. The six samples were separated to four groups, showed the difference of microbial community at different stage during the treatment. Considering the marine microbes found in the river and the dilution of pollutants, tidal water was also involved in the development of microbial community. Therefore, the physiochemical factors and pollutants of the urban river worked together to improved determine in the this microbial river. community structure in the urban river. Figure Figure 6. 6. Canonical Canonical correspondence correspondence analysis analysis (CCA) (CCA) of of microbial microbial community community and and physiochemical physiochemical properties in the river.
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In summary, MBT changed the physical and chemical conditions of the river mainly by increasing the concentration of DO, and then the aerobic microbes were stimulated and activated at degradation of pollutants. Thus, the microbial community would shift and the water quality was improved in this river.
3.5. Microbial Function during MBT Treatment
The microbial community functions were predicted by COG in Figure 7. According to the result, the COG categories related to T (Signal transduction mechanisms), M (Cell wall/membrane/envelope biogenesis), N (Cell motility), J (Translation, ribosomal structure), P (Inorganic ion transport and metabolism), H (Coenzyme transport and metabolism), C (Energy production and conversion), U (Intracellular trafficking, secretion), and F (Nucleotide transport and metabolism) were more abundant before MBT treatment. This might because that the urban black-odor river with high concentration of COD and NH4+-N was suitable for microbes to grow and reproduce, which was treatment matched with could the mitigate high microbial the pollution abundance of river. of sample d1. Figure Figure 7. 7. Functional Functional groups groups by by COG COG categories categories showed showed the the difference difference of microbial of microbial function function before before and after MBT treatment in the river (The left was the abundance of different functional groups; the middle was the percentage of functional group abundance with 95% confidence intervals; the p value was shown in the rightmost; p < 0.05, means significant difference).
Meanwhile microbes responsible to K (transcription), E (Amino acid transport and metabolism), G (carbohydrate transport and metabolism), I (lipid transport and metabolism) and Q (secondary metabolites biosynthesis) were enriched after MBT treatment. It has been reported that the secondary metabolism are well known for the ability to inhibit other microorganisms [40]. Additionally, the result
Water 2019, 11, 66 12 of 14
might due to the reduction of nutrient in the river after MBT treatment, the microorganisms had to compete for resources. The changes of microbial community function showed that the MBT treatment could mitigate the pollution of river.
4. Conclusions
MBT was applied to mitigate the pollution of an urban black-odor river in South China. After the MBT treatment, the concentration of DO was increased and the microbial community was changed, which led to improvement of water quality in the river. The result indicated that MBT is suitable for in situ treatment to mitigate the pollution of the urban black-odor river. Microbial community was shifted during the treatment, and aerobic microbes were dominant at the end of treatment. The relative abundance of dominant classes and genera were changed with the physiochemical conditions and pollution degree in the process of MBT treatment. Thus, MBT could be a possible solution to counteract the deterioration of urban rivers in South China before starting an interception project.
Supplementary Materials: The following are available online at http://www.mdpi.com/2073-4441/11/1/66/s1, Figure S1: Monthly mean temperature of Dongguan City (2016–2018), Figure S2. The Concentration of TP (mg/L) during remediation, Table S1. The ACE and Chao 1 index of samples.
Author Contributions: Y.W. has conceived the study. She also carried out the experiment, data collection, analysis of data, and prepared the first edition of the manuscript. S.L., Y.H., H.L., W.Y. and S.S. participated in the design of the study and helped to draft and edit the manuscript. All authors read and approved the final manuscript.
Funding: This work was supported by National Natural Science Foundation of Guangdong, China (2016A030313136), the National Natural Science Foundation of China (No. 21607006 and No. 51878170), and the Special Financial Grant from the China Postdoctoral Science Foundation (No. 212400227).
Conflicts of Interest: The authors declare no conflicts of interest.
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Fantastic Properties of Nano-bubbles
Kaneo Chiba Masayoshi Takahashi
Burst at the surface
Goes up rapidly
Ordinary bubble (Macrobubble)
3
Disappear Shrinking
Microbubble
Continue
Nano-bubble
Waste water treatment
Nanobubble production
Collapse
Physical stimulus
Microbubble
Collapse
Physical stimulus
Nano-bubble production by collapsing microbubble
・OHDecomposition
of organic chemicals
Radical generation
Microbubble
・OH
with salt (electrolytes) Nano-bubble
4
Nono-bubble generator
Microbubble Nano-bubble
Physical stimulus
5
Analysis of nano-bubble by Dynamic light scattering (DLS)
02468101214161820
76 93 113 138 168 204 249 304 370
Particle size (nm)
6
The deference of size between MB and NB
Microbubble 10 ~30μm
Nano-bubble
< 200nm
7
The deference of life time between MB and NB
Disappear
Continue
Shrinking
Microbubble Nanobubble
Several minutes Several months
Why ? 8
Electrode ( + ↔ − ) Light Electrode
(− ↔ +)
Electrophoresis cell
Microscope
Water + microbubble
Movement of microbubbles in electrophoresis cell
9
Change in ζ potential of microbubble over time
Accumulation of surrounding ions
-80
-70
-60
-50
-40
-30
-20
0 5 10 15 20 25 30
Diameter (μm)
time Ions
Ions Microbubble
with salt (electrolytes)
time
10
Stabilization mechanism of nanobubble (assumption)
Negatively charged
Aqueous solution
Surrounded by accumulated ions
(counter ions)
Salting out
Prevention of gas dispersion
11
+
+
+
+ +
+
+
+
--- +-
-
--- ++
+++-
-
++ ++
++
Nano-bubble
Nano-bubble generator
12
Normal method
Ozone
Bubbling
Nano-bubble
Collapse
Micro-bubble Nano-bubble
13
Normal ozone water vs. Ozone nano-bubble
Ozone water ; disinfection & sterilizing (Food industry, medical )
0.1mg/L 1.5mg/L
1 hour
1.5mg/L
More than 1.0mg/L
100 days
Norovirus
Norovirus symptoms:
vomiting, diarrhea, a low-grade fever, chills, headache, muscle aches
How to treat the problem of norovirus How to treat the problem of norovirus
14
Oyster
During cultivation
Conventional method
Oyster
Ozone water Oyster
Ozone = strong oxidation ability
↓ Virus → inactivation Norovirus 15
Norovirus
Chlorine
Chlorine ( high concentration )
----- not good for foods
Oysters in ozone nanobuble water for 1 day
16
Before ozone nanobubble treatment
17
After ozone nanobubble treatment
More than 99% of feline calicivirus in the body --- inactivated
18
Practical method for inactivation of norovirus by ozone nano-bubble 19
Active effect for fishes
Carps : recovering from Carp herpes ( serious disease )
20
Oxidation ability of ozone nano-bubble as a form of mist
21
more than 80% of original oxidation ability
reduction of smell and bacteria from atmospheres 22
Collapse of oxygen microbubble About 1% salt
Recovery of weakened fishes
Active effect for creatures
23
Oxygen nanobubble
Nano-Bubble Aquarium 24
Astonishing effects on plant
Orchid in oxygen NB water for more than 1 month
Still alive
25
Initial growth of plants
White radish
White radish Broccoli Broccoli
Tap water Oxygen nano-bubble water
26
Active effect for oysters
-20°C for 1 day in oxygen NB
Room temperature
After 1 hour
Still alive
27
Application of nanobubble in medicine
Prevention of arteriosclerosis (hardening of blood vessel )
Oxygen nanobubble inhibits mRNA expression induced by TNFα in RAOECs
ICAM1
VCAM1
GAPDH
Culture aorta endothelium of rat Cytokine stimulation(TNF alpha)
Heart attack
Stroke
28
control alpha TNF
control alpha
TNF
With oxygen nanobubble
Department of Cardiology, Jichi Medical University
Application of nanobubble in medicine
Preservation of living tissue / organ
Vagus nerve of rat : after 1 week
Isotonic NaCl solution Oxygen nanobubble
Tokyo Medical and Dental University
29
30 Thank you very much
NACTE
National Association of Concerned Veterans 4850 Jay Street NE, Washington, DC 20019 202 680-3142
------------------------------------------------------
CREDENTIALS: Service History & Experience
Summary
NACV (National Association of Concerned Veterans) - Since 1978 we have been providing a wide range of services to all veterans who honorably served our country from all branches of the military.
NACV has been training and employing veterans since its inception but made this a major focus in the early 70s through the GI EDucational Bill, CETA, and even the CASE book attempts to provide college credit, certifications and credentialing assistance for military training and experience. NACV began as community-based self-help vets helping vets direct services organization so we knew if we needed jobs we needed to create them if we needed services we needed to provide them if we needed financial assistance we needed to generate it.
NACV provides: comprehensive readjustment transition and re-entry services to vets and their family’s discharge upgrade and compensation and claims benefit assistance.
Jobs creation and jobs training to include: on the job training, WFD, DOES, CWT
Vocational (VOC) rehab and education services
Homeless and housing assistance, training employment and job readiness/placement assistance
GE Program model PHP, IOP and credit classes for transitioning vets
Substance abuse and recovery programs.
PTSD and all other DSM and ICD 10 codes and diagnoses focusing on military and combat-related disorders
Specific Program Experience
Comprehensive, proactive program creation, management & administration.
CETA, VAworkstudy, and college work-study helped ease the plight of recently discharged Veterans. There were 1500 plus VA certified schools around the country and after 74 NAVPA member institutions, there were thousands of jobs created facilitated by VCIP program funds: veterans cost of instruction Program funding created by The Congress through HEW. these were monies paid to colleges universities and post-secondary institutions to provide services for vets per capita vet enrollment.
it funded GED classes, tutoring, child care, discharge review and upgrades, some comp and claim assistance, emergency assistance, employment planning, and placement, temp services, and morphing into small business incubators and micro enterprises, even things like housing for student vets, utilities asst, deposits and assist, and later into housing rehab, vets buying a four-unit building and fixing it up one room and one unit at a time. and rent out rooms etc. and even the continuation of the Catch 22 supply sgt and military loans etc.
NACV was heavily involved in the creation of a vets office for vet entrepreneurs and small businesses, the DOL creation of Veterans Employment and training services (VETS) and ultimately OSDBU program,
NACV received contracts to train and employ vets via SBA, Energy, CSA, DOL, EPA, Commerce From 78-84 NACV train, certified and employed literally 100s of DC area vets in haz mat, asbestos abatement, lead abatement, cable television construction and installation, weatherizaton, energy conservation, transportation, small business incubators, housing construction, remodeling, renovation, housing management, thrift and estate sales, housing furniture, hauling and disposal, moving, demolition, landscaping, and all aspects of construction: electric, plumbing, hvac, roofing, masonry, flooring, insulation, windows doors, siding, framing, carpentry, tile work, cement and paving, brick, drywall, painting, plaster, taping, finishing, auto mechanics, small engine repair, restoration, grounds keeping, IT, smart wiring, deconstruction,
NACV worked with the homeless programs from their inception during The Vietnam era there was a time when one out of three or one out of four new admissions to prison or homeless were vets 16-30 unemployment rate for vets esp in urban inner cities.
NACV work collaboratively with a number of Vietnam vet groups around the country swords to plowshares, there were few allies in the beginning because the older vets did not accept the Vietnam vets A
From 84 to 94 NACV completed literally hundreds of projects in the DMV alone primarily in housing, asbestos, and lead abatement and govt contracts.
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