Supplementary Materials. Microbial Communities in Liquid and Fiber Fractions of Food Waste Digestates

Transcription

1 Supplementary Materials Journal: Applied Microbiology and Biotechnology Microbial Communities in Liquid and Fiber Fractions of Food Waste Digestates are differentially resistant to inhibition by ammonia Wei Peng 1, 2, Fan Lü 1, 2, *, Liming Shao 2, 3, Pinjing He 2, 3 1. State Key Laboratory of Pollution Control & Resource Reuse, Tongji University, Shanghai , PR China 2. Institute of Waste Treatment and Reclamation, Tongji University, Shanghai , PR China 3. Centre for the Technology Research and Training on Household Waste in Small Towns & Rural Area, Ministry of Housing and Urban-Rural Development of PR China (MOHURD) * Corresponding author. Tel/fax: addresses: lvfan.rhodea@tongji.edu.cn. 1

2 Table S1. Bacterial ARISA length in different batches. Bacterial S-1 S-3 S-5 S-7 L-1 L-3 L-5 L-7 bands (bp)

3 Table S2. Archaeal ARISA length in different batches. Archaeal S-1 S-3 S-5 S-7 L-1 L-3 L-5 L-7 bands (bp)

4 PLFA total concentration (mol/l) PLFA PLEL PLEL total concentration (mol/l) 0 S-initial S1 S3 S5 S7 L-initial L1 L3 L5 L7 0 Fig S1. Total concentration of PLFAs and PLEL of different batches. Error bars represent the data range. 4

5 Method S1. Physical and chemical analysis methods Gas composition (CH4 and CO2) was analyzed using a gas chromatograph equipped with a thermal conductivity detector (GC9800, Shanghai Kechuang Co., Ltd, China). Gas pressure in the serum bottles was measured by a Testo 512 manometer to calculate biogas production. Liquid samples were analyzed for ph, volatile fatty acids (VFAs) and total organic carbon (TOC). ph was measured immediately after sampling using a ph meter (phs-2f, Shanghai Precision and Scientific Instrument Co., Ltd, China). Liquid samples were passed through polyester filters (0.22 μm pore size) before the measurement of VFAs and TOC. VFAs (including formate, acetate, propionate, n-butyrate, iso-butyrate, n-valerate and iso-valerate) were determined by high-performance liquid chromatography (LC-20AD, Shimadzu, Japan). TOC was measured with a total organic carbon meter (TOC-V CPH, Shimadzu, Japan) using the 68 C combustion catalytic oxidation/nondispersive infrared (NDIR) method. TAN was measured by an Automatic Kjeltec Nitrogen Analyzer (Kjeltec 8400TM, Foss, Denmark). Free ammonia nitrogen (FAN) was calculated as (Eq.(1)): NH 3 Ka TAN [ H ] Ka 1 [ H ] (1) Where [NH3] is the concentration (mg/l) of free ammonia; [TAN] is the concentration (mg/l) of total ammonia; Ka is the temperature-dependent dissociation constant ( at 35 C); and [H] is the concentration of hydrogen ion (equal to 10 ph ). 5

6 Method S2. Measurement of PLFA and PLEL by GC-MS A GC-MS instrument was equipped with an HP-5 capillary column (50 m 0.2 mm ID; coated with a cross-linked 5% phenylmethyl rubber phase, film thickness 0.3 μm). The carrier gas was He and the flow rate was 1 ml/min. For PLFA analysis, the temperature was increased after 2 min from the initial temperature of 120 C to 205 C at a rate of 10 C/min, kept at 205 C for 1 min, increased from 205 C to 260 C at a rate of 5 C/min then kept at 260 C for 20 min. For PLEL analysis, the temperature was increased after 2 min from the initial temperature of 70 C to 130 C at a rate of 20 C/min, increase from 130 C to 320 C at a rate of 4.0 C/min then kept at 320 C for 20 min. Method S3. DNA extraction, ARISA analysis and PCA Total DNA was extracted using a Powersoil TM DNA isolation kit (MO-BIO Laboratories Inc., CA). Automated ribosomal intergenic spacer analysis (ARISA), which is a fingerprint technique, was used to explore the bacterial and archaeal profiles after different treatments, and was done on a Bioanalyzer (Agilent 2100). Gel-like images of ARISA were converted to decimal data using the software Quantity One Version (Bio-Rad, USA) to obtain the band intensity in each lane. The density and position of the bands were subjected to principal component analysis (PCA) using the PAST software (Hammer et al. 2001). Reference Hammer O, Harper DAT, Ryan PD (2001) PAST: Paleontological statistics software package for education and data analysis. Palaeontologla Electronica 4:1-9. 6