Herbert H. P. Fang

Effect of pH on hydrogen production from glucose by a mixed culture

The effect of pH on the conversion of glucose to hydrogen by a mixed culture of fermentative bacteria was evaluated. At 36 degrees C, six hours hydraulic retention, over 90% of glucose was degraded at pH ranging 4.0-7.0, producing biogas and an effluent comprising mostly fatty acids. At the optimal pH of 5.5, the biogas comprised 64 +/- 2% of hydrogen with a yield of 2.1 +/- 0.1 mol-H2/mol-glucose and a specific production rate of 4.6 +/- 0.4 l-H2/(g-VSS day). The effluent was composed of acetate (15.3-34.1%) and butyrate (31.2-45.6%), plus smaller quantities of other volatile fatty acids and alcohols. The diversity of microbial communities increased with pH, based on 16S rDNA analysis by denaturing gradient gel electrophoresis (DGGE).

Extraction of extracellular polymeric substances (EPS) of sludges.

The efficacies of extracting extracellular polymeric substances (EPS) from aerobic, acidogenic and methanogenic sludges using EDTA, cation exchange resin and formaldehyde under various conditions were compared. Results show that formaldehye plus NaOH was most effective in extracting EPS for all sludges; only 1.1-1.2% of DNA in the sludge samples were detected, suggesting the EPS extracted were not contaminated by intracellular substances. For each gram of volatile solids, formaldehyde-NaOH extracted 165, 179 and 102 mg of EPS from aerobic, acidogenic and methanogenic sludges, respectively. All EPS were mainly composed of carbohydrate, protein and humic substance, plus small quantities of uronic acid and DNA. Carbohydrate was predominant in the acidogenic sludge (62% in the EPS extracted by formaldehyde-NaOH), whereas protein was predominant in the methanogenic sludge (41%). Humic substance, which has often been overlooked, accounted for 30.6, 8.4 and 22.8% of the extracted EPS from aerobic, acidogenic and methanogenic sludges, respectively. However, judging from EPS quantities estimated from confocal laser scanning microscopic observations, formaldehyde-NaOH extracted only a limited portion of EPS. Optimization of extraction procedures and/or development of a more effective extraction method are warranted.

Fermentative hydrogen production from wastewater and solid wastes by mixed cultures

Over 160 publications related to fermentative hydrogen production from wastewater and solid wastes by mixed cultures are compiled and analyzed. Of the 98 reported cases, 57 used single substrates (mainly carbohydrates), 8 used actual wastewater, and 33 used solid wastes for hydrogen conversion. The key information is compiled in four tables: (1) pretreatment conditions for screening hydrogen-producing bacteria from anaerobic sludge or soil, and the process and performance parameters for (2) single substrates in synthetic wastewaters, (3) actual wastewaters, and (4) solid wastes. Process parameters discussed include pH, temperature, hydraulic retention time, seed sludge, nutrients, inhibitors, reactor design, and the means used for lowering hydrogen partial pressure. Performance parameters discussed include hydrogen yield, maximum volumetric production rate, maximum specific production rate, and conversion efficiency. The outlook for this new technology is discussed at the end.

Antibiotic resistance genes in water environment

The use of antibiotics may accelerate the development of antibiotic resistance genes (ARGs) and bacteria which shade health risks to humans and animals. The emerging of ARGs in the water environment is becoming an increasing worldwide concern. Hundreds of various ARGs encoding resistance to a broad range of antibiotics have been found in microorganisms distributed not only in hospital wastewaters and animal production wastewaters, but also in sewage, wastewater treatment plants, surface water, groundwater, and even in drinking water. This review summarizes recently published information on the types, distributions, and horizontal transfer of ARGs in various aquatic environments, as well as the molecular methods used to detect environmental ARGs, including specific and multiplex PCR (polymerase chain reaction), real-time PCR, DNA sequencing, and hybridization based techniques.

Influences of extracellular polymeric substances (EPS) on flocculation, settling, and dewatering of activated sludge

Results in over 200 publications related to constituents, quantities, and characteristics of extracellular polymeric substances (EPS) in activated sludge and their influences on sludge flocculation, settling, and dewatering are compiled and analyzed. Constituents and quantities of EPS are strongly dependent on the extraction process. Most results show that EPS in activated sludge comprise substantially more proteins than carbohydrates. EPS and sludge flocs carry negative zeta potentials and negative charge. Sludge hydrophobicity increases with protent content of the EPS, and an increased hydrophobicity generally leads to a better flocculation. Most data show that sludge settles less well, as measured by sludge volume index, with increased amount of EPS. Sludge settling and dewatering characteristics are affected by the zeta potential, surface charge, and hydrophobicity, as well as the presence of multivalent mineral cations. Process conditions of activated sludge such as sludge retention time, dissolved oxygen, and nutrient levels affect the EPS properties and, thus, the sludge flocculation and settling characteristics. The effects of EPS quantities, ratio of protein and carbohydrate in EPS, sludge digestion, mineral cations, chelating reagent, nutrient level, and pretreatment on sludge dewaterability are discussed.

Biohydrogen production from starch in wastewater under thermophilic condition.

Batch experiments were conducted to convert starch in wastewater into hydrogen at 55 degrees C at various wastewater pH (4.0-9.0) and starch concentrations (9.2-36.6 g/l). The maximum hydrogen yield of 92 ml/g of starch added (17% of the theoretical value) was found at wastewater pH 6.0, and the maximum specific hydrogen production rate of 365 ml/(g-VSS.d) was at wastewater pH 7.0. The methane-free biogas contained up to 60% of hydrogen. The mixed liquor was composed mostly of acetate (40.2-53.4%) and butyrate (26.0-40.9%). Phylogenetic analysis based on 16S rDNA sequences of the 72 clones developed from the sludge at pH 6.0 shows that 85.7% of the clones were closely affiliated with genus Thermoanaerobacterium in family Thermoanaerobacteriaceae; the remaining 14.3% were with an uncultured Saccharococcus sp. clone ETV-T2.

Applications of real-time polymerase chain reaction for quantification of microorganisms in environmental samples

Due to the advanced development of fluorogenic chemistry, quantitative real-time polymerase chain reaction (qRT-PCR) has become an emerging technique for the detection and quantification of microorganisms in the environment. Compared with the conventional hybridization- and PCR-based techniques, qRT-PCR not only has better sensitivity and reproducibility, but it is also quicker to perform and has a minimum risk of amplicon carryover contamination. This article reviews the principle of this emerging technique, its detection reagents, target DNAs, quantification procedures, and affecting factors. The applications of qRT-PCR for the quantification of microorganisms in the environment are also summarized.

Effects of toxic metals and chemicals on biofilm and biocorrosion

Microbes in marine biofilms aggregated into clusters and increased the production of extracellular polymeric substances (EPS), by over 100% in some cases, when the seawater media containing toxic metals and chemicals, such as Cd(II), Cu(II), Pb(II), Zn(II), AI(III), Cr(III), glutaraldehyde, and phenol. The formation of microbial cluster and the increased production of EPS, which contained 84-92% proteins and 8-16% polysaccharides, accelerated the corrosion of the mild steel. However, there was no quantitative relationship between the degree of increased corrosion and the toxicity of metals/chemicals towards sulfate-reducing bacteria, or the increased EPS production.

Quantification of bacterial adhesion forces using atomic force microscopy (AFM).

This study demonstrated that atomic force microscopy (AFM) can be used to obtain high-resolution topographical images of bacteria, and to quantify the tip-cell interaction force and the surface elasticity. Results show that the adhesion force between the Si3N4 tip and the bacteria surface was in the range from -3.9 to -4.3 nN. On the other hand, the adhesion forces at the periphery of the cell-substratum contact surface ranged from -5.1 to -5.9 nN and those at the cell-cell interface ranged from -6.5 to -6.8 nN. The two latter forces were considerably greater than the former one, most likely due to the accumulation of extracellular polymer substance (EPS). Results also show that the elasticity varied on the cell surface.

THE ROLES OF CALCIUM IN SLUDGE GRANULATION DURING UASB REACTOR START-UP

Six upflow anaerobic sludge blanket (UASB) reactors were concurrently operated for 146 d to examine the effects of calcium on the sludge granulation process during start-up. Introduction of Ca2+ at concentrations from 150 to 300 mg/l enhanced the biomass accumulation and granulation process. The calcium concentration in the granules was nearly proportional to the calcium concentration in the feed, and calcium carbonate was the main calcium precipitate in the granules. The specific activity of granules decreased with increasing influent calcium concentration. The optimum calcium concentration for the granulation was from 150 to 300 mg/l. The addition of low-concentration calcium to the UASB reactors appeared to enhance the three steps of sludge granulation: adsorption, adhesion and multiplication, but it did not lead to a different proliferation of predominant microorganisms in the granules.