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Frontiers of Environmental Science & Engineering

ISSN 2095-2201

ISSN 2095-221X(Online)

CN 10-1013/X

Postal Subscription Code 80-973

2018 Impact Factor: 3.883

Front. Environ. Sci. Eng.    2017, Vol. 11 Issue (1) : 10
Isolation and application of predatory Bdellovibrio-and-like organisms for municipal waste sludge biolysis and dewaterability enhancement
Ran Yu(),Shiwen Zhang,Zhoukai Chen,Chuanyang Li
Key Laboratory of Energy Thermal Conversion and Control (Ministry of Education), Department of Environmental Science and Engineering, School of Energy and Environment, Southeast University, Nanjing 210096, China
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Indigenous predatory BALO strains were successfully isolated from activated sludge.

Sludge SRF and CST were significantly reduced by BALOs induced biolysis process.

The increase of BALO input dosage promoted the sludge biolysis efficiency.

Sludge biolysis disintegrated flocs and lysed cells for internal water release.

The optimal sludge biolysis time was 24 h and no pH adjustment was needed.

Bdellovibrio-and-like organisms (BALOs) are a group of ubiquitous and obligate predatory bacteria and commonly used as biocontrol agents. In this study, an efficient, environmental-friendly, and convenient BALOs encouraged municipal waste sludge biolysis pretreatment technique was developed and investigated for dewaterability enhancement of excess waste sludge. The indigenous predatory BALOs were successfully isolated from the sludge for biolysis treatment. Without any chemical addition or pH adjustment, the sludge specific resistance (SRF) and capillary suction time (CST) were significantly reduced by as high as 53.4% and 23.8%, respectively within 24 h’s treatment, which would further be lowered with the increase of BALOs input dosage. However, the continuous extension of reaction time would worsen the sludge dewaterability. The decreases of SRF and CST accompanied with the increases of sludge disintegration degree and soluble chemical oxygen demand, nitrogen, and phosphorus concentrations all emphasized the contributions of BALOs’ predation activities to sludge disturbance, cell lysis, and consequently the release of sludge intracellular water to finally effectively improve the sludge dewaterability and disposal efficiency.

Keywords Bdellovibrio-and-like organisms (BALOs)      Biolysis      Activated sludge      Dewaterability      Predation     
Corresponding Authors: Ran Yu   
Issue Date: 09 January 2017
 Cite this article:   
Ran Yu,Shiwen Zhang,Zhoukai Chen, et al. Isolation and application of predatory Bdellovibrio-and-like organisms for municipal waste sludge biolysis and dewaterability enhancement[J]. Front. Environ. Sci. Eng., 2017, 11(1): 10.
sample No. reduction rate/% sample No. reduction rate/% sample No. reduction rate/%
D1 71.7±0.7 D11 64.1±1.2 D21 80.6±3.0
D2 80.9±0.2 D12 70.4±0.63 D22 67.8±1.6
D3 75.4±1.0 D13 63.4±1.0 D23 45.9±0.4
D4 68.9±1.3 D14 71.6±2.5 D24 49.6±0.8
D5 47.7±0.4 D15 65.2±2.2 D25 46.5±0.7
D6 66.1±1.2 D16 48.5±0.7 D26 70.0±1.9
D7 40.9±0.5 D17 39.3±0.6 D27 48.8±0.8
D8 90.9+ 3.0 D18 82.9±3.7 D28 76.9±1.9
D9 73.4±0.8 D19 39.9±0.4 D29 86.2±0.5
D10 44.5±0.3 D20 69.1±1.7 D30 68.8±1.6
Tab.1  The reduction of OD600 during BALOs’ liquid enrichments co-incubated with (1.0±0.1) × 109 CFU•mL-1 prey culture
Fig.1  D8 BALO strain enrichments with different prey species
group strain number SRF reduction rate/%
12 h 24 h 36 h 48 h
1 control-1 15.8±2.8 16.6±5.7 6.8±2.8 -9.8±4.2
D1 13.5±2.8 8.6±2.8 4.3±1.4 -5.3±1.4
D2 33.5±1.4 39.2±3.1 29.5±2.8 -1.9±3.4
D3 24.7±7.1 41.6±4.2 30.0±5.7 14.8±5.6
D4 23.6±5.7 41.7±2.8 23.4±1.4 5.6±2.8
D6 35.8±2.8 38.5±7.1 34.7±4.2 21.4±1.4
2 control-2 -1.5±4.7 -1.8±5.4 5.8±5.7 -12.6±2.8
D8 -14.6±5.1 4.6±4.2 6.8±4.2 -18.6±3.1
D9 4.8±2.4 6.3±2.8 17.6±1.4 -1.1±4.7
D11 -35.3±6.1 -47.4±4.2 2.5±2.6 -22.7±3.4
D12 7.7±5.7 13.6±3.7 18.1±3.3 1.5±5.7
D13 6.9±7.1 18.1±5.7 17.8±4.4 2.6±2.1
3 control-3 19.2±4.2 15.2±2.8 1.3±2.5 -3.0±1.2
D14 6.1±8.5 30.7±5.7 33.1±2.8 30.3±2.8
D15 23.0±4.2 47.2±1.7 39.4±3.8 25.5±4.1
D18 27.7±5.1 43.9±2.5 38.1±2.6 21.1±5.7
D20 23.8±5.7 40.7±3.7 38.2±7.1 -3.9±3.9
D21 28.7±2.8 35.6±1.8 38.3±1.4 28.4±3.7
4 control-4 4.5±3.7 2.4±1.4 -7.0±5.7 -15.0±7.1
D22 23.0±3.4 16.7±5.7 9.8±6.4 3.7±5.7
D26 5.8±7.1 4.2±7.1 11.1±2.8 -0.5±4.2
D28 6.2±5.3 20.0±4.6 0.4±4.2 -4.3±3.4
D29 30.8±2.8 9.2±6.2 5.7±4.2 -0.1±5.0
D30 -0.1±4.2 -3.0±4.2 -7.8±3.6 -18.4±2.8
Tab.2  The variations of SRF during the BALO-sludge interaction processes
Fig.2  The predation performances of the five BALO strains during their co-incubations with Klebsiella-1 ((1.2±0.1) × 109 CFU•mL-1), respectively (a) and the SRF variation profiles of the sludge lysed by the five BALO cultures of the similar concentrations (6.2±0.1) × 106 PFU·mL-1) in the sludges, respectively (b)
Fig.3  The impacts of sludge pH on SRF during the sludge biolysis with D15 culture ((4.8±0.3) × 106 PFU·mL-1)
Fig.4  The effects of BALO input dosage on sludge dewatering performance improvement in terms of SRF (a) and CST (b) (Note: the legends indicate the magnitudes of the initial D15 concentrations in the sludge samples which were (3.6±0.6) × 106 PFU·mL-1 ((1.4±0.2) × 108 PFU·g MLSS-1), (6.4±0.6) × 107 PFU·mL-1 ((2.6±0.2) × 109 PFU·g MLSS-1), and (4.6±0.6) × 108 PFU·mL-1 ((1.8±0.2) × 1010 PFU·g MLSS-1), respectively)
Fig.5  The variations of MLSS concentration and MLVSS/MLSS ratio (a), DDSCOD and SCOD concentrations (b), and SP, SN, and NH3-N concentrations (c), and the relationship between DDSCOD and SRF or CST (d) during the sludge biolysis in the presence of (4.2±0.3) × 107 PFU•mL-1 D15 culture
Fig.6  ESEM images of the control sludge sample (a) and the one (b) treated with D15 for 24 h with 1,000 fold of magnification. The images in the oval white frame was the further enlargement of the area pointed by the arrow with 3,000 fold of magnification
1 Vesilind P A, Hsu C C. Limits of sludge dewaterability. Water Science and Technology, 1997, 36(11): 87–91
2 Mowla D, Tran H N, Allen D G. A review of the properties of biosludge and its relevance to enhanced dewatering processes. Biomass and Bioenergy, 2013, 58: 365–378
3 Huo M, Zheng G, Zhou L. Enhancement of the dewaterability of sludge during bioleaching mainly controlled by microbial quantity change and the decrease of slime extracellular polymeric substances content. Bioresource Technology, 2014, 168(3): 190–197 pmid: 24650613
4 Tsang K R, Vesilind P A. Moisture distribution in sludges. Water Science and Technology, 1990, 22(12): 135–142
5 Chen G, Yue P L, Mujumdar A S. Sludge dewatering and drying. Drying Technology, 2002, 20(4&5): 883–916
6 Dincler A, Vesilind P A. Effect of sludge water distribution on the liquid–solid separation of a biological sludge. Journal of Environmental Science and Health. Part A, Environmental Science and Engineering & Toxic and Hazardous Substance Control, 2003, 38(10): 2391–2400
7 Ferrentino R, Langone M, Merzari F, Tramonte L, Andreottola G. A review of anaerobic side-stream reactor for excess sludge reduction: configurations, mechanisms, and efficiency. Critical Reviews in Environmental Science and Technology, 2016, 46(4): 382–405
8 Sockett R E. Predatory lifestyle of Bdellovibrio bacteriovorus. Annual Review of Microbiology, 2009, 63(1): 523–539 pmid: 19575566
9 El-Shanshoury A E R R, Abo-Amer A E, Alzahrani O M. Isolation of Bdellovibrio sp. from wastewater and their potential application in control of Salmonella paratyphi in water. Geomicrobiology Journal, 2016, 33(10): 886–893
10 Fry J C, Staples D G. Distribution of Bdellovibrio bacteriovorus in sewage works, river water, and sediments. Applied and Environmental Microbiology, 1976, 31(4): 469–474
pmid: 1267445
11 Özkan M, Çelik M A, Karagöz P, Yılmaz H, Şengezer Ç. Activity of Bdellovibrio on sludge bacteria and its potential use for cleaning of membrane bioreactors. New Biotechnology, 2014, 31S: S133
12 Jurkevitch E, Minz D, Ramati B, Barel G. Prey range characterization, ribotyping, and diversity of soil and rhizosphere Bdellovibrio spp. isolated on phytopathogenic bacteria. Applied and Environmental Microbiology, 2000, 66(6): 2365–2371 pmid: 10831412
13 Johnke J, Cohen Y, de Leeuw M, Kushmaro A, Jurkevitch E, Chatzinotas A. Multiple micro-predators controlling bacterial communities in the environment. Current Opinion in Biotechnology, 2014, 27: 185–190 pmid: 24598212
14 Dwidar M, Monnappa A K, Mitchell R J. The dual probiotic and antibiotic nature of Bdellovibrio bacteriovorus. BMB Reports, 2012, 45(2): 71–78 pmid: 22360883
15 Withey S, Cartmell E, Avery L M, Stephenson T. Bacteriophages--potential for application in wastewater treatment processes. Science of the Total Environment, 2005, 339(1-3): 1–18 pmid: 15740754
16 Christensen M L, Keiding K, Nielsen P H, Jørgensen M K. Dewatering in biological wastewater treatment: a review. Water Research, 2015, 82: 14–24 pmid: 25959073
17 Dias F F, Bhat J V. Microbial ecology of activated sludge II. Bacteriophages, Bdellovibrio, Coliforms, and other organisms. Applied and Environmental Microbiology, 1965, 13(2): 257–261
pmid: 14325890
18 Shapiro O H, Kushmaro A, Brenner A. Bacteriophage predation regulates microbial abundance and diversity in a full-scale bioreactor treating industrial wastewater. ISME Journal, 2010, 4(3): 327–336 pmid: 19924159
19 Kadouri D, O’Toole G A. Susceptibility of biofilms to Bdellovibrio bacteriovorus attack. Applied and Environmental Microbiology, 2005, 71(7): 4044–4051 pmid: 16000819
20 Medina A A, Kadouri D E. Biofilm formation of Bdellovibrio bacteriovorus host-independent derivatives. Research in Microbiology, 2009, 160(3): 224–231 pmid: 19223013
21 Eaton A D, Clesceri L S, Rice E W, Greenberg A E, Franson M A H, eds. Standard methods for the examination of water and wastewater. 21 ed: Washington, DC: APHA, AWWA and WEF, 2005
22 Huan L, Yiying J, Mahar R B, Zhiyu W, Yongfeng N. Effects of ultrasonic disintegration on sludge microbial activity and dewaterability. Journal of Hazardous Materials, 2009, 161(2-3): 1421–1426 pmid: 18547717
23 Nadkarni M A, Martin F E, Jacques N A, Hunter N. Determination of bacterial load by real-time PCR using a broad-range (universal) probe and primers set. Microbiology, 2002, 148(Pt 1): 257–266 pmid: 11782518
24 Varon M, Shil M. Interacton of Bdellovibrio bacteriovorus and host bacteria. I. Kinetic studies of attachment and invasion of Escherichia coli B by Bdellovibrio bacteriovorus. Journal of Bacteriology, 1968, 95(3): 744–753
pmid: 4868362
25 Schwarzenbeck N, Borges J M, Wilderer P A. Treatment of dairy effluents in an aerobic granular sludge sequencing batch reactor. Applied Microbiology and Biotechnology, 2005, 66(6): 711–718 pmid: 15558277
26 Jenkins D, Richard M G, Daigger G. Manual of the Control of Activated Sludge Bulking and Foaming. 2 ed. Michigan: Lewis Publisher, 1993
27 Pineiro S A, Sahaniuk G E, Romberg E, Williams H N. Predation pattern and phylogenetic analysis of Bdellovibrionaceae from the Great Salt Lake, Utah. Current Microbiology, 2004, 48(2): 113–117 pmid: 15057478
28 Markelova N Y. Predacious bacteria, Bdellovibrio with potential for biocontrol. International Journal of Hygiene and Environmental Health, 2010, 213(6): 428–431 pmid: 20850380
29 Eskicioglu C, Kennedy K J, Droste R L. Characterization of soluble organic matter of waste activated sludge before and after thermal pretreatment. Water Research, 2006, 40(20): 3725–3736 pmid: 17028065
30 More T T, Yan S, Tyagi R D, Surampalli R Y. Potential use of filamentous fungi for wastewater sludge treatment. Bioresource Technology, 2010, 101(20): 7691–7700 pmid: 20542684
31 Schoeffield A J, Williams H N, Turng B, Fackler W A Jr. A comparison of the survival of intraperiplasmic and attack phase bdellovibrios with reduced oxygen. Microbial Ecology, 1996, 32(1): 35–46 pmid: 8661540
32 Pasternak Z, Njagi M, Shani Y, Chanyi R, Rotem O, Lurie-Weinberger M N, Koval S, Pietrokovski S, Gophna U, Jurkevitch E. In and out: an analysis of epibiotic vs. periplasmic bacterial predators. ISME Journal, 2014, 8(3): 625–635 pmid: 24088628
33 Varon M, Shilo M. Interaction of Bdellovibrio bacteriovorus and host bacteria. II. Intracellular growth and development of Bdellovibrio bacteriovorus in liquid cultures. Journal of Bacteriology, 1969, 99(1): 136–141
pmid: 4895842
34 Dashiff A, Keeling T G, Kadouri D E. Inhibition of predation by Bdellovibrio bacteriovorus and Micavibrio aeruginosavorus via host cell metabolic activity in the presence of carbohydrates. Applied and Environmental Microbiology, 2011, 77(7): 2224–2231 pmid: 21317250
35 Bougrier C, Carrère H, Delgenès J P. Solubilisation of waste-activated sludge by ultrasonic treatment. Chemical Engineering Journal, 2005, 106(2): 163–169
36 Davidov Y, Friedjung A, Jurkevitch E. Structure analysis of a soil community of predatory bacteria using culture-dependent and culture-independent methods reveals a hitherto undetected diversity of Bdellovibrio-and-like organisms. Environmental Microbiology, 2006, 8(9): 1667–1673 pmid: 16913926
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