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

1. Reaktör Meclisi <ol><li> Şekil 1&#39;de şematik göre reaktör parçaları düzenleyin. <ol><li> Bir karıştırma plaka üzerinde reaktör (R) koyun reaktöre bir karıştırma çubuğu ekleyin. Tankın çıkış (üst) bağlantı noktası laboratuarlarında kenarına doğru yönlendirilir ve böylece heyecan plaka ve reaktörün yanında geri dönüşüm tankı (RT) yerleştirin. </li><li> Geri dönüşüm tankı (RT) çıkış (üst) port altına atık kabını (W) yerleştirin. Geri dönüşüm tankı (RT) sonraki besleme tankı (FT) yerleştirin. Not: Besleme tankı 5 L. toplam kapasitesi </li></ol></li><li> Uygun büyüklükte bir standı ve kelepçe ile devrilmeye karşı reaktör (R) sabitleyin. Aynı şekilde, hareket etmesini önlemek için geri dönüşüm tankı (RT) sabitleyin. </li><li> Recycle (Pompa A) neopren peristaltik pompa hortumunu takın ve (Pompa B) pompa kafaları beslenirler. Ek boru spesifikasyonları için malzemeler tablosuna bakın. Vidalar yazarlara ile pompa sürücüler üzerine pompa kafaları yükleyinPompa sürücülerle ded. </li><li> Reaktör ve geri dönüşüm tankı üzerinde limanlarına Pompa A&#39;nın boru bağlayın. Besleme tankı ve geri dönüşüm tankına Pompa B&#39;nin hortumun ucunu. Boru ile geri dönüşüm deposuna üst reaktör noktasını bağlayın. Reaktör limanlarında tüp kıskaçları uygulayın. Not: Fotosentetik topluluklar lambalar tarafından sağlanan yapay aydınlatma yararlanabilir. </li></ol> Stok Solutions, Akan / Yem Çözümleri ve Yosun Aşılayıcı 2. Hazırlık <ol><li> Mineral stok çözelti hazırlayın. 200 g sodyum bikarbonat, 40 g monobazik potasyum fosfat, 4 g magnezyum sülfat, 4 g demir klorür, 4 g kalsiyum klorür, 1 g bakır klorür, 1 g kobalt: deiyonize su, 500 ml bir 1 L&#39;lik bir volumetrik balona aşağıdakileri ekleyin klorür hekzahidrat, 1 g nikel klorür heksahidrat, 1 g çinko sülfat heptahidrat. Iyonu giderilmiş su ilave 400 ml ilave edilir. Tuzların çözülmesini teşvik etmek zorla girdap. Aşağıdaki disstuzların ların çözüme, 1 L. çözeltisinin toplam hacmi getirmek için iyonu giderilmiş su ilave </li><li> Amonyak stok çözelti hazırlayın. 1 L&#39;lik bir volumetrik bir şişede, iyonu giderilmiş su, yaklaşık 900 ml amonyum klorit 38,214 gram çözülür. Dağılmasından sonra, 1000 ml toplam hacmi getirmek için deiyonize su ilave edilir. Not: 1 L&#39;ye seyreltilmiş stok çözeltisi 1 ml 10 mg L-verir -1 NH4 + -N çözeltisi. </li><li> Nitrat stok çözelti hazırlayın. 1 L&#39;lik bir volumetrik bir şişede, iyonu giderilmiş su, yaklaşık 900 ml potasyum nitrat 72,413 g çözülür. Dağılmasından sonra, 1000 ml toplam hacmi getirmek için deiyonize su ilave edilir. Not: - N çözeltisi 1 L&#39;ye seyreltilmiş stok çözeltisi 1 mi -1 Resim 3, bir 10 mg L-verir. </li><li> Besleme / akın çözüm hazırlayın. Bir 2 ml sulandırmak, N - 20 mg L içeren bir besleme solüsyonu -1 NH 4 + N ve 20 mg L-1 NO 3 yapmak içinmmonia stok çözeltisi ve 1 L&#39;lik toplam hacme kadar nitrat stok çözeltisi 2 mi. Seyreltmeden öne, 0.5 ml mineral çözeltisi / çözeltinin L yapılan ekleyin. Reaktörü başlatmak için toplamda akan 5 L hazırlayın. </li><li> Yosun aşı malzemesi hazırlayın. <ol><li> Böyle bir akarsu veya gölet gibi bir yosun içeren su vücuttan su büyük bir hacmi (en az 10 L) toplayın. Alg 24 saat rahatsız su örnekleri bırakarak yerleşmek için izin verin. </li><li> Numune şişeleri içinde konsantre yosun süspansiyonu bırakarak, Durusu ve örneklerin üstündeki net (içermeyen yosun) su atın. Bir konteyner içine numunelerin tüm alg süspansiyon birleştirin ve çökmesini ve şişeden adımları tekrarlayın. </li><li> Konsantre numune içinde biyokütle ölçün. <ol><li> Co ölçmek oda sıcaklığında 30 dakika boyunca bir kurutucuda soğutmadan sonra 103 ° C&#39;ye ayarlanmış bir fırın içinde tekne O / N ağırlık kağıt vakum filtresi (0.45 um MCE vakum filtresi) ve alüminyum KuruFiltrenin kitle mbined ve tekne tartın. </li><li> Vakum konsantre yosun süspansiyonu 20 ml filtresi ve filtre dönmek ve / N O kurumaya fırın tekne tartın. </li><li> Filtrenin kombine kitle ölçün ve tekne tartın. Konsantre numune içinde biyokütle yoğunluğunu hesaplar. Not: Araştırmacılar kaynak suyu vücudun bağlıdır toplamak gerekir su numunesi toplam hacmi. </li></ol></li></ol></li></ol> 3. Tohum ve Reaktör Başlangıç <ol><li> Reaktöre besleme çözeltisinin 750 ml ilave edilir. Besleme çözeltisinin 500 ml &#39;si ile yeniden doldurun. </li><li> Yavaşça reaktörün tabanına yakın yosun 1.5 g ihtiva eden bir süspansiyon inoküle eklemek için uzun bir pipet kullanın. Aşı bir sonraki aşamaya geçmeden önce, reaktörün dibine çöker görsel gözlem ile Bunu sağlamak için izin verir. </li><li> Hücreler yerleşmiş sonra, (10 Revolutio tüp kelepçelerini çıkarın ve yavaş akış hızına Pompa A açmakns min -1 / 38 ml dk -1). Hortumun içinde sıkışıp Hava reaktörüne ihraç edilecektir. Not: reaktöre 750 ml ilave reaktörü terk pompa tarafından rahatsız herhangi bir biyokütle önleyecektir. Tüm hava sınırdışı edilmiş olmasını sağlamak için boru sıkın. </li><li> Çözelti, reaktör içine pompalanır Yavaş yavaş devri daim tankı besleme çözeltisi ilave edilir. Reaktör ve geri dönüşüm deposu hem kapasite ve üst port üzerinden geri dönüşüm tankı çıkmak için atık başlar olana kadar eklenmesini devam edin. Not: besleme çözeltisinin hacmi reaktöre ilâve edilen inokulantın hacmine bağlı olarak değişecektir devri daim tankı eklenmesi. </li><li> Besleme tankına geri kalan besleme solüsyonu dökün. </li><li> 72.5 ml dk bir geri dönüşüm akış hızı kuran, -1 dk 19 devrimlere geri dönüşüm pompası (Pompa A) Set -1. Yosun reaktörün tabanından loft başlar dikkate alınmalıdır. Reaktörün üzerinde geçişleri kullanarak, yosun bi belirlemekomass bölge yüksekliği. Yükseklik sonraki adıma geçmeden önce sabit olduğundan emin olun. </li><li> Çok düşük hızlarda karıştırma plakası açın; 1 ya da 2 ayarı başlatmak için uygun değildir. Karıştırma çubuğu ayrıca biyokütle lofting yardımcı olacaktır, ama agresif karıştırma, reaktör bırakın geri dönüşüm tankı girin ve atık bırakmak için yosun neden olur. Reaktör (Şekil 2A) içinde net bir alg sınır kurmak için gerekli bir ayar hızını karıştırma ayarlayın; alg biyokütle bölgesi yüksekliği yaklaşık 10-15 cm olmalıdır. </li><li> Alg fiş ve reaktör sıvısı arasında net bir sınır gözlemledikten sonra besleme pompasını başlatın. 1,5 ml dakika akış oranı oluşturulması, 1 dak 25 devir Pompayı -1. Nedeniyle gelen içeri giren akımının neden olduğu ağırlık ve yer değiştirmesine reaktör sıvı çıkış portu çıkış gözlemleyin. </li></ol> 4. Numune Toplama ve Analizi <ol><li> Numune toplama faaliyetleri pri yürütmekveya reaktör sistemi üzerinde bakım yapmadan için. Günlük atık ve içeri giren numunelerin 20 ml toplayın. Geri dönüşüm tankı içinde gelen atık örnekleri toplayın. Besleme tankının doğrudan içeri giren örneklerini toplayın. </li><li> Vakum filtre örnekleri depolama ve analiz öncesinde askıda katı kaldırmak için. </li><li> Daha analize kadar -20 ° C &#39;de ve dışarıya akanların örnekleri saklayın. Numuneler maruz kalan donma çözülme döngüsü sayısını sınırlayın. Gerekirse, örnekler örnek bütünlüğünü korumak için kısma bölünebilir. </li><li> Standart teknikler kullanılarak 17 nitrat, nitrit, amonyak ve örnek analizi yürütmek. Not: İyon Kromatografi (IC) kullanılan yazarlar burada sunulan sonuçlar üretmek için. Şartnamede için malzemeler tablosuna bakın. </li></ol>

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M., Eveillard, D., Tonon, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+metabolic+approach+to+study+algal-bacterial+interactions+in+changing+environments.">A metabolic approach to study algal-bacterial interactions in changing environments.</a> Mol Ecol. 23 (7), 1656-1660 (2014).</li><li>Watanabe, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Symbiotic+association+in+Chlorella+culture.">Symbiotic association in Chlorella culture.</a> FEMS Microbiol Ecol. 51 (2), 187-196 (2005).</li><li>Burke, C., Thomas, T., Lewis, M., Steinberg, P., Kjelleberg, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Composition,+uniqueness+and+variability+of+the+epiphytic+bacterial+community+of+the+green+alga+Ulva+australis.">Composition, uniqueness and variability of the epiphytic bacterial community of the green alga Ulva australis.</a> ISME J. 5 (4), 590-600 (2011).</li><li>Krohn-Molt, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metagenome+survey+of+a+multispecies+and+alga-associated+biofilm+revealed+key+elements+of+bacterial-algal+interactions+in+photobioreactors.">Metagenome survey of a multispecies and alga-associated biofilm revealed key elements of bacterial-algal interactions in photobioreactors.</a> Appl Environ Microbiol. 79 (20), 6196-6206 (2013).</li><li>Cooper, E. D., Bentlage, B., Gibbons, T. R., Bachvaroff, T. R., Delwiche, C. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metatranscriptome+profiling+of+a+harmful+algal+bloom.">Metatranscriptome profiling of a harmful algal bloom.</a> Harmful Algae. 37, 75-83 (2014).</li></ol>

The authors have nothing to disclose and declare that they have no competing financial interests.

Bu bölüm olası operasyonel konularda yanı sıra farklı mikrobiyal topluluklar kullanarak çözmek için gerekli protokol varyasyonları bir tartışma başlayacak. Bu reaktör tasarımının güçlü oksijen akı kontrolü ve reaktör içinde yüksek yoğunluklu flocs oluşumunu yönetme yeteneği dahil olmak üzere, ele alınacaktır. Güncel sorunlar ve soruşturmanın olası caddeleri de söz edilecektir. Protokol nüansları ve çeşitleri Kültürlerin farklı yeti?...

Kentsel atık su genellikle askıda katı madde (SS), biyolojik oksijen talebi (BOD), organik ve inorganik azot ve fosfor içeriği 5,6 azaltmak amacıyla aktif çamur işlemleri ile muamele edilir. Aktif çamur yöntemi, sekonder atık su işleme için bir araç, gelen atık ve geri dönüşümlü heterotrofik mikroorganizmanın karışık sıvısı ile doldurulmuş bir havalandırma tankında organik karbon oksidasyonunu gerektirir 5-7 (yaygın aktif çamur olarak da adlandırılır). Karışık likör sonra çamur için, daha kolay toplanması için yerleşir nispeten büyük arıtıcı (dinlendirme tankı) girer elden çıkarılması veya açıklık, arıtılmış atık su üçüncül tedavi veya dezenfeksiyon devam edebilirsiniz ederken salınan önce, geri havalandırma tankına geri dönüştürülebilir ya alan sular 5-7. İkincil arıtıcı tedavi atıksu ve katı (çamur) Verimli ayrılması oldu düzgün fonksiyonu için gereklidirtewater arıtma sistemi, herhangi bir aktif çamur atık 5-8 BOİ ve SS artacak Çöktürme ötesine devam olarak. Alternatif biyolojik süreçlerin bir dizi azaltmak veya ekli-büyüme (biyofilm) reaktörler, membran Biyoreaktörlerin (MBR&#39;leri) ve granül çamur reaktörler de dahil olmak üzere geniş açıklığa kavuşturulması tankları, ihtiyacını ortadan kaldırmak atıksuyun ikincil tedavisi için mevcuttur. Biyofilm reaktör, doğal agrega mikroorganizmalar ve katı bir yüzey üzerinde bir katman olarak bağlandıkları biyofilmlerin oluşumuyla olarak, açıklık tankın gerek kalmadan biyokütle tutma ve birikimi sağlar. Biyofilm reaktörler üç tip halinde sınıflandırılabilir: paketlenmiş yataklı reaktörler, akışkan yataklı reaktörler ve dönen biyolojik. Böyle bir trickling filtreleri ve biyolojik kuleleri Paketli yataklı reaktörler, sabit bir katı büyüme yüzeyi 5,6 kullanmaktadır. Akışkan yataklı reaktörler, (FBRs), parçacıkların Mikroorganizmaların bağlanması bağlıdırkum gibi granül karbon (GAC) ya da yüksek yukarı doğru akış oranı 9,10 ile süspansiyon içinde muhafaza edilmiştir cam boncuk, aktif. Döner biyolojik reaktörler biyofilm sağlayan bir döner şaftına bağlı ortam üzerinde oluşturulmuş biyofilm alternatif olarak havaya maruz kalma ve sıvı 5,6 tedavi edilen bağlıdır. MBR&#39;leri biyoreaktör (batık konfigürasyonunda) içinde veya dışarıdan devridaim (yan-akım konfigürasyonu) 5,11 vasıtasıyla ya, membran filtrasyon üniteleri kullanın. Membranlar tedavi sıvı 11,12 den biyokütle ve katı parçacıkların iyi ayrılmasını sağlamak için hizmet vermektedir. Granül çamur reaktörler içinde mikroorganizmaların çok yoğun ve iyi yerleşme granüllerin oluşumu yüksek yüzeysel hava yukarı doğru çıktığı maruz kaldığında ortaya çıkar 13 hızlar yukarı akış reaktörlerdir. Aktif çamur işlemi, yeni bir yukarı akış reaktör sistemi için başka bir alternatif olarak, hemen yüksek yoğunluklu biyoreaktör (HDBR) olarak adlandırılan, designe oldud ve yoksul yerleşim çamur oluşmasına neden olduğu bilinmektedir düşük F / M koşullarında sentetik atık akışları (yani, kabarma çamur) 1,7,14 dan aktif çamur tarafından KOİ giderimi incelemek için Satış ve (2006) Shieh tarafından inşa. Kullanılan HDBR sistemi tipik olarak, bir yukarı akış reaktörü ve bir dış devri daim tank, akışkan yataklı reaktörler modifiye. Biyofilm kaplı substrat tutulacak şekilde Akışkan yataklı reaktörler tipik olarak yeterince süspansiyon haline biyofilm büyüme alt katmanı tutmak için yeterince yüksek, fakat düşük yeniden çevrim akımı, akış oranları ile çalıştırılır. Akışkan yataklı reaktörlerde aksine, HDBR Satış anlatılan ve (2006) Shieh dış havalandırma ile birlikte, reaktörün 1 bünyesinde oluşturulan biyokütle bölgenin bozulması önlenir, nispeten düşük devri daim akımı debileri kullanılır. Daha sonraki çalışmalar başarıyla bakteri 3,4 denitrifikasyon / nitrifikasyon kullanarak azot akışlarının bir dizi tedavi etmek için bu reaktör tasarımın yeteneğini göstermiştir. Tüm saplamaHDBR içinde istikrarlı, yoğun biyokütle bölgenin oluşumu harici flokülasyon / sedimantasyon süreci 1-4 ihtiyacını ortadan ies. Burada rapor olarak yoğun kültürleri büyümeye HDBR kullanımı da alg yetiştirilmesi için bir foto-biyo (PBR) konfigürasyonunda olarak test edilmiştir. Biz yararları ve alg ekimi için bu yeni reaktör sisteminin sakıncalarını ve biyokütle toplama (yani, iyi katı-sıvı ayırma 15,16) ile ilişkili Alg biyoyakıt ticarileştirilmesi büyük bir engel üstesinden gelmek için potansiyel tartışmak. Aşağıdaki protokol başlatma, örnek, monte ve ilgi mikrobiyal topluluk olarak yosun ile HDBR korumak için gerekli adımları açıklar. Heterotrofik ve nitrifikasyon / Azot salan kültürlerinin başlangıç ​​ve işletme protokolü değişimler de söz edilecektir. Son olarak, genel avantajları, dezavantajları, ve bu yeni reaktör tasarımının bilinmeyenler vurgulanır.

<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.

40 mg -NL -1 beslemedeki toplam nitrojen içeriğine korurken HDBR, içeri giren amonyak ve sodyum nitrat konsantrasyonlarında çeşitli oranları üzerinden yosun yetiştirmek için kullanıldı. Akın ve atık numuneleri günlük alındı; biyokütle yoğunluğu numuneleri her durumun başında ve sonunda alındı. Reaktör koşulları değiştirildi sonra kararlı durum dengeye ulaşmak için ortalama 3-5 gün sürdü. Içeri giren koşulları geniş bir yelpazede üzerinde belirgin bir biyokütle bölges...

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.

Çeşitli mikrobiyal Toplulukları Yüksek Yoğunluklu Yetiştirme için Yeni Bir Biyoreaktör

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

a-novel-bioreactor-for-high-density-cultivation-diverse-microbial

Research

JoVE Journal

Bioengineering

16.8K Görüntüleme.  Drexel University. 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.

Video: A Novel Bioreactor for High Density Cultivation of Diverse Microbial Communities

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

Complex tissue culture matrices, in which types and concentrations of biological stimuli (e.g. growth factors, inhibitors, or small molecules) or matrix structure (e.g. composition, concentration, or stiffness of the matrix) vary over space, would enable a wide range of investigations concerning how these variables affect cell differentiation, migration, and other phenomena. The major challenge in creating layered matrices is maintaining the structural integrity of layer interfaces without diffusion of individual components from each layer1. Current methodologies to achieve this include photopatterning2-3, lithography4, sequential functionalization5, freeze drying6, microfluidics7, or centrifugation8, many of which require sophisticated instrumentation and technical skills. Others rely on sequential attachment of individual layers, which may lead to delamination of layers9. 
DGMP overcomes these issues by using an inert density modifier such as iodixanol to create layers of varying densities10. Since the density modifier can be mixed with any prepolymer or bioactive molecule, DGMP allows each scaffold layer to be customized. Simply varying the concentration of the density modifier prevents mixing of adjacent layers while they remain aqueous. Subsequent single step polymerization gives rise to a structurally continuous multilayered scaffold, in which each layer has distinct chemical and mechanical properties. The density modifier can be easily removed with sufficient rinsing without perturbation of the individual layers or their components. This technique is therefore well suited for creating hydrogels of various sizes, shapes, and materials.
A protocol for fabricating a 2D-polyethylene glycol (PEG) gel, in which alternating layers incorporate RGDS-350, is outlined below. We use PEG because it is biocompatible and inert. RGDS, a cell adhesion peptide11, is used to demonstrate spatial restriction of a biological cue, and the conjugation of a fluorophore (Alexa Fluor 350) enables us to visually distinguish various layers. This procedure can be adapted for other materials (e.g. collagen, hyaluronan, etc.) and can be extended to fabricate 3D gels with some modifications10.

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

Here we describe a unique strategy for creating biocompatible, layered matrices with continuous interfaces between distinct layers for tissue engineering. Such a scaffold could provide an ideal customizable environment to modulate cell behavior by various biological, chemical or mechanical cues

Yoğunluk Gradient Katmanlı Polimerizasyonu (DGMP): Multi-bölmesi oluşturma için A Novel Tekniği, Doku Mühendisliği için Özelleştirilebilir İskelesi

Complex tissue culture matrices, in which types and concentrations of biological stimuli (e.g. growth factors, inhibitors, or small molecules) or matrix ...

Biyomühendislik

Design of a Biaxial Mechanical Loading Bioreactor for Tissue Engineering

We designed a loading device that is capable of applying uniaxial or biaxial mechanical strain to a tissue engineered biocomposites fabricated for transplantation. While the device primarily functions as a bioreactor that mimics the native mechanical strains, it is also outfitted with a load cell for providing force feedback or mechanical testing of the constructs. The device subjects engineered cartilage constructs to biaxial mechanical loading with great precision of loading dose (amplitude and frequency) and is compact enough to fit inside a standard tissue culture incubator. It loads samples directly in a tissue culture plate, and multiple plate sizes are compatible with the system. The device has been designed using components manufactured for precision-guided laser applications. Bi-axial loading is accomplished by two orthogonal stages. The stages have a 50 mm travel range and are driven independently by stepper motor actuators, controlled by a closed-loop stepper motor driver that features micro-stepping capabilities, enabling step sizes of less than 50 nm. A polysulfone loading platen is coupled to the bi-axial moving platform. Movements of the stages are controlled by Thor-labs Advanced Positioning Technology (APT) software. The stepper motor driver is used with the software to adjust load parameters of frequency and amplitude of both shear and compression independently and simultaneously. Positional feedback is provided by linear optical encoders that have a bidirectional repeatability of 0.1 &mu;m and a resolution of 20 nm, translating to a positional accuracy of less than 3 &mu;m over the full 50 mm of travel. These encoders provide the necessary position feedback to the drive electronics to ensure true nanopositioning capabilities. In order to provide the force feedback to detect contact and evaluate loading responses, a precision miniature load cell is positioned between the loading platen and the moving platform. The load cell has high accuracies of 0.15% to 0.25% full scale.

We designed a novel mechanical loading bioreactor that can apply uniaxial or biaxial mechanical strain to a cartilage biocomposite prior to transplantation into an articular cartilage defect.

Doku Mühendisliği için iki eksenli Mekanik Yükleniyor Biyoreaktör tasarımı

We  designed a loading device that is capable of applying uniaxial or biaxial  mechanical strain to a tissue engineered biocomposites fabricated for  ...

Microfluidic Picoliter Bioreactor for Microbial Single-cell Analysis: Fabrication, System Setup, and Operation

In this protocol the fabrication, experimental setup and basic operation of the recently introduced microfluidic picoliter bioreactor (PLBR) is described in detail. The PLBR can be utilized for the analysis of single bacteria and microcolonies to investigate biotechnological and microbiological related questions concerning, e.g.&#160;cell growth, morphology, stress response, and metabolite or protein production on single-cell level. The device features continuous media flow enabling constant environmental conditions for perturbation studies, but in addition allows fast medium changes as well as oscillating conditions to mimic any desired environmental situation. To fabricate the single use devices, a silicon wafer containing sub micrometer sized SU-8 structures served as the replication mold for rapid polydimethylsiloxane casting. Chips were cut, assembled, connected, and set up onto a high resolution and fully automated microscope suited for time-lapse imaging, a powerful tool for spatio-temporal cell analysis. Here, the biotechnological platform organism Corynebacterium glutamicum&#160;was seeded into the PLBR and cell growth and intracellular fluorescence were followed over several hours unraveling time dependent population heterogeneity on single-cell level, not possible with conventional analysis methods such as flow cytometry. Besides insights into device fabrication, furthermore, the preparation of the preculture, loading, trapping of bacteria, and the PLBR cultivation of single cells and colonies is demonstrated. These devices will add a new dimension in microbiological research to analyze time dependent phenomena of single bacteria under tight environmental control. Due to the simple and relatively short fabrication process the technology can be easily adapted at any microfluidics lab and simply tailored towards specific needs.

In this protocol the fabrication, setup and basic operation of a microfluidic picoliter bioreactor (PLBR) for single-cell analysis of prokaryotic microorganisms is introduced. Industrially relevant microorganisms were analyzed as proof of principle allowing insights into growth rate, morphology, and phenotypic heterogeneity over certain time periods, hardly possible with conventional methods.

In this protocol the fabrication, experimental setup and basic operation of the recently introduced microfluidic picoliter bioreactor (PLBR) is described ...

Operation of a Benchtop Bioreactor

Fermentation systems are used to provide an optimal growth environment for many different types of cell cultures. The ability afforded by fermentors to carefully control temperature, pH, and dissolved oxygen concentrations in particular makes them essential to efficient large scale growth and expression of fermentation products. This video will briefly describe the advantages of the fermentor over the shake flask. It will also identify key components of a typical benchtop fermentation system and give basic instruction on setup of the vessel and calibration of its probes. The viewer will be familiarized with the sterilization process and shown how to inoculate the growth medium in the vessel with culture. Basic concepts of operation, sampling, and harvesting will also be demonstrated. Simple data analysis and system cleanup will also be discussed.

Fermentors are used to increase culture yield and productivity of bioengineered cells. After screening multiple microbial or animal cell culture candidates in shake flasks, the next logical step is to increase the selected culture&#x2019;s biomass with the fermentor. This video demonstrates the setup and operation of a typical benchtop bioreactor system.

Bir Benchtop Biyoreaktör Operasyonu

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

Construction and Characterization of a Novel Vocal Fold Bioreactor

In vitro engineering of mechanically active tissues requires the presentation of physiologically relevant mechanical conditions to cultured cells. To emulate the dynamic environment of vocal folds, a novel vocal fold bioreactor capable of producing vibratory stimulations at fundamental phonation frequencies is constructed and characterized. The device is composed of a function generator, a power amplifier, a speaker selector and parallel vibration chambers. Individual vibration chambers are created by sandwiching a custom-made silicone membrane between a pair of acrylic blocks. The silicone membrane not only serves as the bottom of the chamber but also provides a mechanism for securing the cell-laden scaffold. Vibration signals, generated by a speaker mounted underneath the bottom acrylic block, are transmitted to the membrane aerodynamically by the oscillating air. Eight identical vibration modules, fixed on two stationary metal bars, are housed in an anti-humidity chamber for long-term operation in a cell culture incubator. The vibration characteristics of the vocal fold bioreactor are analyzed non-destructively using a Laser Doppler Vibrometer (LDV). The utility of the dynamic culture device is demonstrated by culturing cellular constructs in the presence of 200-Hz sinusoidal vibrations with a mid-membrane displacement of 40 &#181;m. Mesenchymal stem cells cultured in the bioreactor respond to the vibratory signals by altering the synthesis and degradation of vocal fold-relevant, extracellular matrix components. The novel bioreactor system presented herein offers an excellent in vitro platform for studying vibration-induced mechanotransduction and for the engineering of functional vocal fold tissues.

A novel vocal fold bioreactor capable of delivering physiologically relevant, vibratory stimulation to cultured cells is constructed and characterized. This dynamic culture device, when combined with a fibrous poly(&#949;-caprolactone) scaffold, creates a vocal fold-mimetic environment that modulates the behaviors of mesenchymal stem cells.

Bir Roman Vokal Fold Bioreactor İnşaatı ve Karakterizasyonu

In vitro engineering of mechanically active tissues requires the presentation of physiologically relevant mechanical conditions to cultured cells. To ...

Cultivation of Mammalian Cells Using a Single-use Pneumatic Bioreactor System

Recent advances in mammalian, insect, and stem cell cultivation and scale-up have created tremendous opportunities for new therapeutics and personalized medicine innovations. However, translating these advances into therapeutic applications will require in vitro systems that allow for robust, flexible, and cost effective bioreactor systems. There are several bioreactor systems currently utilized in research and commercial settings; however, many of these systems are not optimal for establishing, expanding, and monitoring the growth of different cell types. The culture parameters most challenging to control in these systems include, minimizing hydrodynamic shear, preventing nutrient gradient formation, establishing uniform culture medium aeration, preventing microbial contamination, and monitoring and adjusting culture conditions in real-time. Using a pneumatic single-use bioreactor system, we demonstrate the assembly and operation of this novel bioreactor for mammalian cells grown on micro-carriers. This bioreactor system eliminates many of the challenges associated with currently available systems by minimizing hydrodynamic shear and nutrient gradient formation, and allowing for uniform culture medium aeration. Moreover, the bioreactor&#8217;s software allows for remote real-time monitoring and adjusting of the bioreactor run parameters. This bioreactor system also has tremendous potential for scale-up of adherent and suspension mammalian cells for production of a variety therapeutic proteins, monoclonal antibodies, stem cells, biosimilars, and vaccines.

Using a pneumatic bioreactor, we demonstrate the assembly, operation, and performance of this single-use bioreactor system for the growth of mammalian cells.

Tek kullanımlık Pnömatik Biyoreaktör Sistemi kullanılarak memeli hücrelerinde yetiştirme

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

Construction and Setup of a Bench-scale Algal Photosynthetic Bioreactor with Temperature, Light, and pH Monitoring for Kinetic Growth Tests

The optimal design and operation of photosynthetic bioreactors (PBRs) for microalgal cultivation is essential for improving the environmental and economic performance of microalgae-based biofuel production. Models that estimate microalgal growth under different conditions can help to optimize PBR design and operation. To be effective, the growth parameters used in these models must be accurately determined. Algal growth experiments are often constrained by the dynamic nature of the culture environment, and control systems are needed to accurately determine the kinetic parameters. The first step in setting up a controlled batch experiment is live data acquisition and monitoring. This protocol outlines a process for the assembly and operation of a bench-scale photosynthetic bioreactor that can be used to conduct microalgal growth experiments. This protocol describes how to size and assemble a flat-plate, bench-scale PBR from acrylic. It also details how to configure a PBR with continuous pH, light, and temperature monitoring using a data acquisition and control unit, analog sensors, and open-source data acquisition software.

This paper describes the assembly process and operation of a bench-scale photosynthetic bioreactor that can be used, in conjunction with other methods, to estimate pertinent kinetic growth parameters. This system continuously monitors the pH, light, and temperature using sensors, a data acquisition and control unit, and open-source data acquisition software.

Kinetik Büyüme Testleri için Sıcaklık, Işık ve pH İzleme ile Bir Bench Ölçekli Algal Fotosentetik Biyoreaktörün Kurulması ve Kurulması

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

Process Optimization using High Throughput Automated Micro-Bioreactors in Chinese Hamster Ovary Cell Cultivation

Optimization of bioprocesses to increase the yield of desired products is of importance in the biopharmaceutical industry. This can be achieved by strain selection and by developing bioprocess parameters. Shake flasks have been used for this purpose. They, however, lack the capability to control the process parameters such as pH and dissolved oxygen (DO). This limitation can be overcome with the help of an automated micro-bioreactor. These bioreactors mimic cultivation at a larger scale. One of the major advantages of this system is the integration of the Design of Experiment (DOE) in the software. This integration enables establishing a design where multiple process parameters can be varied simultaneously. The critical process parameters and optimum bioprocess conditions can be analyzed within the software. The focus of the work presented here is to introduce the user to the steps involved in process design in the software and incorporation of the DOE within the cultivation run.

Here, we present a detailed procedure to run a Design of Experiment in an automated micro-bioreactor followed by cell harvest and protein quantification using a Protein A column.

Çin Hamster Yumurtalık Hücre Yetiştiriciliğinde Yüksek İş-İş BulaMetli Otomatik Mikro-Biyoreaktörler Kullanılarak Proses Optimizasyonu

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

High-Throughput Identification of Resistance to Pseudomonas syringae pv. Tomato in Tomato using Seedling Flood Assay

Tomato is an agronomically important crop that can be infected by Pseudomonas syringae, a Gram-negative bacterium, resulting in bacterial speck disease. The tomato-P. syringae pv. tomato pathosystem is widely used to dissect the genetic basis of plant innate responses and disease resistance. While disease was successfully managed for many decades through the introduction of the Pto/Prf gene cluster from Solanum pimpinellifolium into cultivated tomato, race 1 strains of P. syringae have evolved to overcome resistance conferred by the Pto/Prf gene cluster and occur worldwide.
Wild tomato species are important reservoirs of natural diversity in pathogen recognition, because they evolved in diverse environments with different pathogen pressures. In typical screens for disease resistance in wild tomato, adult plants are used, which can limit the number of plants that can be screened due to their extended growth time and greater growth space requirements. We developed a method to screen 10-day-old tomato seedlings for resistance, which minimizes plant growth time and growth chamber space, allows a rapid turnover of plants, and allows large sample sizes to be tested. Seedling outcomes of survival or death can be treated as discrete phenotypes or on a resistance scale defined by amount of new growth in surviving seedlings after flooding. This method has been optimized to screen 10-day-old tomato seedlings for resistance to two P. syringae strains and can easily be adapted to other P. syringae strains.

High-Throughput Identification of Resistance to Pseudomonas syringae pv. Tomato in Tomato using Seedling Flood Assay

The seedling flood assay facilitates rapid screening of wild tomato accessions for the resistance to the Pseudomonas syringae bacterium. This assay, used in conjunction with the seedling bacterial growth assay, can assist in further characterizing the underlying resistance to the bacterium, and can be used to screen mapping populations to determine the genetic basis of resistance.

Pseudomonas syringae pv Direnci yüksek-Throughput Tanımlama. Fide Sel Asası Kullanarak Domates

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

Generation of Dynamical Environmental Conditions using a High-Throughput Microfluidic Device

Mimicking in vivo environmental conditions is crucial for in vitro studies on complex life machinery. However, current techniques targeting live cells and organs are either highly expensive, like robotics, or lack nanoliter volume and millisecond time accuracy in liquid manipulation. We herein present the design and fabrication of a microfluidic system, which consists of 1,500 culture units, an array of enhanced peristaltic pumps and an on-site mixing modulus. To demonstrate the capacities of the microfluidic device, neural stem cell (NSC) spheres are maintained in the proposed system. We observed that when the NSC sphere is exposed to CXCL in day 1 and EGF in day 2, the round-shaped conformation is well maintained. Variation in the input order of 6 drugs causes morphological changes to the NSC sphere and the expression level representative marker for NSC stemness (i.e., Hes5 and Dcx). These results indicate that dynamic and complex environmental conditions have great effects on NSC differentiation and self-renewal, and the proposed microfluidic device is a suitable platform for high throughput studies on the complex life machinery.

We present a microfluidic system for high throughput studies on complex life machinery, which consists of 1500 culture units, an array of enhanced peristaltic pumps and an on-site mixing modulus. The microfluidic chip allows for the analysis of the highly complex and dynamic micro-environmental conditions in vivo.

Yüksek Verimli Mikroakışkan Cihaz Kullanarak Dinamik ÇevreSel Koşulların Üretilmesi

Mimicking in vivo environmental conditions is crucial for in vitro studies on complex life machinery. However, current techniques targeting live cells and ...