Name: a novel bioreactor for high density cultivation of diverse microbial communities
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Description: Watch this Scientific Journal Video about A Novel Bioreactor for High Density Cultivation of Diverse Microbial Communities at JoVE.com

The authors would like to acknowledge Aspen Walker at the University of Pennsylvania for her assistance in reactor maintenance and sample collection.

1. Reattore Assemblea <ol><li> Disporre i componenti del reattore secondo lo schema di figura 1. <ol><li> Posizionare il reattore (R) su una piastra di miscelazione, aggiungere una ancoretta al reattore. Posizionare il serbatoio di riciclo (RT) accanto alla piastra di agitazione e reattore in modo che la porta effluente (superiore) del serbatoio è diretto verso il bordo del banco di laboratorio. </li><li> Posizionare il contenitore degli scarti (W) sotto la porta di effluente (superiore) del serbatoi...

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Questa sezione inizia con una discussione di variazioni di protocollo necessari per affrontare eventuali questioni operative, nonché utilizzando diverse comunità microbiche. La forza di questo disegno reattore saranno discussi, tra cui la possibilità di governare il controllo del flusso di ossigeno e la formazione di fiocchi alta densità all&#39;interno del reattore. Saranno inoltre menzionate le sfide attuali e le possibili vie di studio. Protocollo sfumature e variazioni <br /...

Reflui urbani è comunemente trattati con processo a fanghi attivi per ridurre i solidi sospesi (SS), richiesta di ossigeno biologica (BOD), azoto organico e inorganico, e fosforo 5,6. Il processo a fanghi attivi, un mezzo di depurazione secondario, comporta l&#39;ossidazione del carbonio organico in una vasca di aerazione riempito con un liquido misto di refluo e riciclati microrganismi eterotrofi (comunemente indicato come fanghi attivi) 5-7. Il liquido misto quindi entra in una relativamente grande chiarificatore (decantatore) dove il fango si deposita per facilitare la raccolta, sia di essere smaltiti o riciclati al serbatoio di aerazione, mentre, acque reflue trattate chiarificata può continuare a trattamento terziario o disinfezione prima di essere rilasciato in acque riceventi 5-7. Separazione efficiente di acque reflue trattate e solidi (fanghi) nel chiarificatore secondario è essenziale per il corretto funzionamento di un erasistema di trattamento tewater, come ogni fanghi attivi continuare al di là dei chiarificatori aumenterà la BOD e SS nell&#39;effluente 5-8. Un certo numero di processi biologici alternative esistono per trattamento secondario delle acque reflue, che riducono o eliminano la necessità di grandi serbatoi chiarificatori, compresi crescita divisoria (biofilm) reattori, bioreattori a membrana (MBR), e reattori fanghi granulari. Nei reattori biofilm, la formazione di biofilm, in cui i microrganismi naturalmente aggregato e collegare come uno strato su una superficie solida, permette di ritenzione biomassa e accumulo senza la necessità di un serbatoio chiarire. Reattori biofilm possono essere classificati in tre tipi: i reattori a letto ricco, reattori a letto fluido, e la rotazione contattori biologici. Reattori a letto pranzo, come un percolatori e torri biologiche, utilizzano un stazionario superficie solida crescita 5,6. Reattori a letto fluidizzato (FBRs) dipendono dalla attacco dei microrganismi di particelle,come sabbia, carbone attivo granulare (GAC), o di perle di vetro, che sono tenuti in sospensione da un alto tasso di flusso verso l&#39;alto 9,10. Rotazione reattori biologici dipendono biofilm formati sulla media associati ad un albero rotante che consentono il biofilm essere alternativamente esposte all&#39;aria e il liquido viene trattata 5,6. MBR utilizzano unità di filtrazione a membrana, sia all&#39;interno del bioreattore (configurazione sommerso) o esternamente tramite ricircolo (configurazione side-stream) 5,11. Le membrane servono per ottenere una buona separazione della biomassa e particelle solide dal liquido trattato 11,12. Reattori fango granulare sono reattori flusso verso l&#39;alto in cui la formazione di granuli estremamente densi e ben sedimentazione di microrganismi si verifica quando sono esposti a elevato flusso verso l&#39;alto dell&#39;aria superficiale velocità vettoriale 13. Come ulteriore alternativa al processo a fanghi attivi, un reattore upflow romanzo, ora chiamato un bioreattore ad alta densità (HDBR), era designed e costruito da vendite e Shieh (2006) per studiare la rimozione COD da fanghi attivi dai flussi di rifiuti sintetici in condizioni di scarsa F / M che sono noti per causare la formazione di cattiva decantazione fanghi (cioè, carica fanghi) 1,7,14. Il sistema utilizzato HDBR modificato reattori a letto fluido che in genere consistono di un reattore flusso verso l&#39;alto e un serbatoio di riciclo esterno. Reattori a letto fluido sono in genere azionati con corrente di riciclo Portate abbastanza alto per mantenere il substrato di crescita biofilm sospeso ma bassi abbastanza in modo che il substrato biofilm coperte viene mantenuta. A differenza di reattori a letto fluido, il HDBR descritto nelle vendite e Shieh (2006) ha usato le portate del flusso di riciclaggio relativamente bassi che, insieme con aerazione esterna, impedito interruzione della zona di biomassa formata all&#39;interno del reattore 1. Studi successivi hanno dimostrato la capacità di questo progetto del reattore di trattare con successo una serie di flussi di azoto utilizzando nitrificanti / batteri denitrificanti 3,4. In tutto stalloneies la formazione di una stabile, zona biomassa densa all&#39;interno HDBR eliminato la necessità di un processo di flocculazione / sedimentazione esterno 1-4. Come riportiamo qui, l&#39;uso del HDBR crescere culture dense è stato anche testato in una configurazione photobioreactor (PBR) alla coltura delle alghe. Discutiamo i vantaggi e gli svantaggi di questo sistema reattore romanzo per la coltivazione delle alghe e le sue potenzialità per superare un grande ostacolo nella commercializzazione di biocarburanti algali associati alla raccolta di biomassa (ad esempio, una buona separazione solido-liquido 15,16). Il protocollo che segue illustra i passaggi necessari per assemblare, avvio, campione, e mantenere un HDBR con alghe come la comunità microbica di interesse. Saranno inoltre menzionate variazioni del protocollo di avvio e il funzionamento delle culture eterotrofi e nitrificazione / denitrificanti. Infine, saranno evidenziati i vantaggi generali, gli svantaggi e le incognite di questo disegno romanzo reattore.

<table border="0" cellpadding="0" cellspacing="0" width="686">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Aeration stone</td>
			<td style="width:96px;">Alita</td>
			<td style="width:119px;">AS-3015C</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Aerator</td>
			<td style="width:96px;">Top Fin</td>
			<td style="width:119px;">Air-1000</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Ammonium chloride</td>
			<td style="width:96px;">Sigma Aldrich</td>
			<td style="width:119px;">A9434</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Anion analysis column</td>
			<td style="width:96px;">Shodex</td>
			<td style="width:119px;">IC SI-52 4E</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:213px;">Beaker (600 mL)</td>
			<td style="width:96px;">Corning Pyrex</td>
			<td style="width:119px;">1000-600</td>
			<td style="width:259px;">Used as mixing vessel (MV). Addition of hose barbs at the bottom and 500 mL levels. Outside diameter of hose barbs 3/8&#34;.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Calcium chloride</td>
			<td style="width:96px;">Sigma Aldrich</td>
			<td style="width:119px;">C5670</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Cation analysis column</td>
			<td style="width:96px;">Shodex</td>
			<td style="width:119px;">IC YS-50</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Cobalt chloride hexahydrate</td>
			<td style="width:96px;">Sigma Aldrich</td>
			<td style="width:119px;">C8661</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Copper chloride</td>
			<td style="width:96px;">Sigma Aldrich</td>
			<td style="width:119px;">222011</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Ferric chloride</td>
			<td style="width:96px;">Sigma Aldrich</td>
			<td style="width:119px;">157740</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:213px;">Filter (vacuum)</td>
			<td style="width:96px;">Fisherbrand</td>
			<td style="width:119px;">09-719-2E</td>
			<td style="width:259px;">0.45 um membrane filter, MCE, 47 mm diameter</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:213px;">Graduated cylinder (1000 mL)</td>
			<td style="width:96px;">Corning Pyrex</td>
			<td style="width:119px;">3025-1L</td>
			<td style="width:259px;">Used as reactor vessel (R). Addition of hose barbs at bottom, 500 mL, and 1 L levels. Outside diameter of hose barbs 3/8&#34;.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">HPLC/IC</td>
			<td style="width:96px;">Shimadzu</td>
			<td style="width:119px;">Prominence</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Magnesium sulfate</td>
			<td style="width:96px;">Sigma Aldrich</td>
			<td style="width:119px;">M2643</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:213px;">Masterflex L/S variable speed drive</td>
			<td style="width:96px;">Masterflex</td>
			<td style="width:119px;">07553-50</td>
			<td style="width:259px;">Drive for recycle and feed pumps (2 needed)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Nickel chloride hexahydrate</td>
			<td style="width:96px;">Sigma Aldrich</td>
			<td style="width:119px;">N6136</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Potassium nitrate</td>
			<td style="width:96px;">Sigma Aldrich</td>
			<td style="width:119px;">P8291</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:213px;">(Monobasic) Potassium phosphate</td>
			<td style="width:96px;">Sigma Aldrich</td>
			<td style="width:119px;">P5655</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Pump head</td>
			<td style="width:96px;">Masterflex</td>
			<td style="width:119px;">07018-20</td>
			<td style="width:259px;">Recycle pump head</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Pump head</td>
			<td style="width:96px;">Masterflex</td>
			<td style="width:119px;">07013-20</td>
			<td style="width:259px;">Feed pump head</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Pump tubing</td>
			<td style="width:96px;">Masterflex</td>
			<td style="width:119px;">6404-18</td>
			<td style="width:259px;">Recycle pump tubing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Pump tubing</td>
			<td style="width:96px;">Masterflex</td>
			<td style="width:119px;">6404-13</td>
			<td style="width:259px;">Feed pump tubing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Sodium bicarbonate</td>
			<td style="width:96px;">Sigma Aldrich</td>
			<td style="width:119px;">S5761</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:213px;">Zinc sulfate heptahydrate</td>
			<td style="width:96px;">Sigma Aldrich</td>
			<td style="width:119px;">Z0251</td>
			<td style="width:259px;"></td>
		</tr>
	</tbody>
</table>

a novel bioreactor for high density cultivation of diverse microbial communities

A novel reactor design, coined a high density bioreactor (HDBR), is presented for the cultivation and study of high density microbial communities. Past studies have evaluated the performance of the reactor for the removal of COD1 and nitrogen species2-4 by heterotrophic and chemoautotrophic bacteria, respectively. The HDBR design eliminates the requirement for external flocculation/sedimentation processes while still yielding effluent containing low suspended solids. In this study, the HDBR is applied as a photobioreactor (PBR) in order to characterize the nitrogen removal characteristics of an algae-based photosynthetic microbial community. As previously reported for this HDBR design, a stable biomass zone was established with a clear delineation between the biologically active portion of the reactor and the recycling reactor fluid, which resulted in a low suspended solid effluent. The algal community in the HDBR was observed to remove 18.4% of total nitrogen species in the influent. Varying NH4+ and NO3- concentrations in the feed did not have an effect on NH4+ removal (n=44, p=0.993 and n=44, p=0.610 respectively) while NH4+ feed concentration was found to be negatively related with NO3- removal (n=44, p=0.000) and NO3- feed concentration was found to be positively correlated with NO3- removal (n=44, p=0.000). Consistent removal of NH4+, combined with the accumulation of oxidized nitrogen species at high NH4+ fluxes indicates the presence of ammonia- and nitrite-oxidizing bacteria within the microbial community.

Il HDBR stato usato per coltivare le alghe in diversi rapporti di concentrazione influente ammoniaca e nitrati, pur mantenendo un contenuto di azoto totale nel mangime a 40 mg -NL -1. Influente e campioni effluenti sono stati presi tutti i giorni; campioni di densità di biomassa sono stati prelevati all&#39;inizio e alla fine di ogni condizione. Il reattore sono voluti in media 3-5 giorni per raggiungere l&#39;equilibrio di stato stazionario dopo sono state modificate le condizioni. Su una vasta gamma di con...

Watch this Scientific Journal Video about A Novel Bioreactor for High Density Cultivation of Diverse Microbial Communities at JoVE.com

A Novel Bioreactor for High Density Cultivation of Diverse Microbial Communities

A novel reactor design, coined a high density bioreactor (HDBR), is presented for the cultivation and study of high density microbial communities. Here, the HDBR is successfully applied in a photobioreactor (PBR) configuration for the study of nitrogen metabolism by a mixed high density algal community.

Un bioreattore Novel per l&#39;alta densità di coltivazione diverse comunità microbiche

A novel reactor design, coined a high density bioreactor (HDBR), is presented for the cultivation and study of high density microbial communities. Past ...

Research

JoVE Journal

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La coltivazione di cellule progenitrici neurali umane in un 3-dimensionale di auto-assemblaggio Peptide Hydrogel

The  influence of 3-dimensional (3D) scaffolds on growth, proliferation and finally  neuronal differentiation is of great interest in order to find new ...

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Density Gradient Multilayered Polymerization (DGMP): A Novel Technique for Creating Multi-compartment, Customizable Scaffolds for Tissue Engineering

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Fermentation systems are used to provide an optimal growth environment for many different types of cell cultures. The ability afforded by fermentors to ...

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Recent advances in mammalian, insect, and stem cell cultivation and scale-up have created tremendous opportunities for new therapeutics and personalized ...

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The optimal design and operation of photosynthetic bioreactors (PBRs) for microalgal cultivation is essential for improving the environmental and economic ...

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Optimization of bioprocesses to increase the yield of desired products is of importance in the biopharmaceutical industry. This can be achieved by strain ...

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Tomato is an agronomically important crop that can be infected by Pseudomonas syringae, a Gram-negative bacterium, resulting in bacterial speck disease. ...