Comparing lactate and glycerol as a single-electron donor for sulfate reduction in fluidized bed reactors

Among the greatest challenges to the full implementation of biological sulfate reduction are the cost and availability of the electron source. With the development of the biofuel industry, new organic substrates have become available. Therefore, this work sought to compare the performance of a sulfidogenic process utilizing either lactate or glycerol as the substrate for sulfate-reducing bacteria (SRB) growth. Although sulfate reduction is energetically more favorable with lactate, glycerol is a less expensive alternative because excess production is forecasted with the worldwide development of the biodiesel industry. Continuous experiments were performed in a fluidized bed (FB) reactor containing activated carbon as a carrier for a mixed bacterial population composed of sulfate-reducing and fermentative bacteria. During the lactate-fed phases, incomplete oxidation of lactate to acetate by SRB was the dominant metabolic pathway resulting in as much as 90 % sulfate reduction and high acetate concentrations (2.7 g L^?1). Conversely, in the glycerol-fed phases, glycerol degradation resulted from syntrophic cooperation between sulfate-reducing and fermentative bacteria that produce butyrate along with acetate (1.0 g L^?1) as oxidation products. To our knowledge, this is the first report of butyrate formation during sulfate reduction in a glycerol-fed continuous-flow reactor. Sulfate concentrations were reduced by about 90 % (from 2,000 to 100–300 mg L^?1) when glycerol was being fed to the reactor. Since the FB reactor was able to stand a change from lactate to glycerol, this reactor is recommended as the preferred option should glycerol be selected as a cost-effective alternative to lactate for continuous sulfate reduction.     

Simultaneous removal of hexavalent chromium and <Emphasis Type="Italic">o</Emphasis>-dichlorobenzene by isolated <Emphasis Type="Italic">Serratia marcescens</Emphasis> ZD-9

Heavy metals–organics mixture pollution is increasingly concerned and simultaneous removal of organic pollutants and heavy metals is becoming significant. In this study, a strain was isolated from the sediment of a tannery effluent outfalls, which can remove o-dichlorobenzene (o-DCB) and Cr(VI) simultaneously. The bacterial isolate was identified as Serratia marcescens by the 16S rRNA gene sequences. The strain removed about 90% of o-DCB and more than 80% of Cr(VI) at the concentration of 1.29 g L^?1 o-DCB and 20 mg L^?1 Cr(VI). In the presence of concomitant pollutant o-DCB, the optimal pH (8.0) and temperature (30 °C) were determined for Cr(VI) removal. Changes of the bacterial cells and intracellular black Cr(III) sediments were observed by the TEM auxiliary analysis. The results of the FTIR spectroscopy analysis indicated that hydroxyl, amide and polysaccharides were involved in the process of Cr(VI) removal.     

Homo-fermentative production of d-lactic acid by Lactobacillus sp. employing casein whey permeate as a raw feed-stock

Casein whey permeate (CWP), a lactose-enriched dairy waste effluent, is a viable feed stock for the production of value-added products. Two lactic acid bacteria were cultivated in a synthetic casein whey permeate medium with or without pH control. Lactobacillus lactis ATCC 4797 produced d-lactic acid (DLA) at 12.5 g l^?1 in a bioreactor. The values of Leudking–Piret model parameters suggested that lactate was a growth-associated product. Batch fermentation was also performed employing CWP (35 g lactose l^?1) with casein hydrolysate as a nitrogen supplement in a bioreactor. After 40 h, L. lactis produced 24.3 g lactic acid l^?1 with an optical purity >98 %. Thus CWP may be regarded as a potential feed-stock for DLA production.     

Effect of culture conditions, cytokinins, methyl jasmonate and salicylic acid on the biomass accumulation and production of withanolides in multiple shoot culture of Withania somnifera (L.) Dunal using liquid culture

The influence of cytokinins and culture conditions including medium volume, harvest time and elicitation with abiotic elicitors (SA/MeJ) have been studied for the optimal production of biomass and withanolides in the multiple shoot culture of Withania somnifera. Elicitation of shoot inoculum mass (2 g l^?l FW) with SA at 100 μM in the presence of 0.6 mg l^?l BA and 20 mg l^?l spermidine for 4 h exposure time at the 4th week in 20 ml liquid medium recorded higher withanolides production (withanolides A [8.48 mg g^?l DW], withanolides B [15.47 mg g^?l DW], withaferin A [29.55 mg g^?l DW] and withanone [23.44 mg g^?l DW]), which were 1.14 to 1.18-fold higher than elicitation with MeJ at 100 μM after 5 weeks of culture. SA-elicited cultures did not exhibit much variation in biomass accumulation when compared to control. This cytokinin induces and SA-elicited multiple shoot culture protocol provides a potential alternative for the optimal production of biomass and withanolides utilizing liquid culture.     

Effects of stationary phase elongation and initial nitrogen and phosphorus concentrations on the growth and lipid-producing potential of Chlorella sp. HQ

Higher lipid production and nutrient removal rates are the pursuing goals for synchronous biodiesel production and wastewater treatment technology. An oleaginous alga Chlorella sp. HQ was tested in five different synthetic water, and it was found to achieve the maximum biomass (0.27 g L^?1) and lipid yield (41.3 mg L^?1) in the synthetic secondary effluent. Next, the effects of the stationary phase elongation and initial nitrogen (N) and phosphorus (P) concentrations were investigated. The results show that the algal characteristics were affected apparently under different N concentrations but not P, which were verified by Logistic and Monod models. At the early stationary phase, the algal biomass, lipid and triacylglycerols (TAGs) yields, and P removal efficiency increased and reached up to 0.19 g L^?1, 46.7 mg L^?1, 14.3 mg L^?1, and 94.3 %, respectively, but N removal efficiency decreased from 86.2 to 26.8 % under different N concentrations. And the largest TAGs yield was only 6.4 mg L^?1 and N removal efficiency was above 71.1 % under different P concentrations. At the late stationary phase, the maximal biomass, lipid and TAGs yields, and P removal efficiencies primarily increased as the initial N and P concentrations increase and climbed up to 0.49 g L^?1, 99.2 mg L^?1, 54.0 mg L^?1, and 100.0 %, respectively. It is concluded that stationary phase elongation is of great importance and the optimal initial N/P ratio should be controlled between 8/1 and 20/1 to serve Chlorella sp. HQ for better biodiesel production and secondary effluent purification.     

Performance and granulation in an upflow anaerobic sludge blanket (UASB) reactor treating saline sulfate wastewater

An upflow anaerobic sludge blanket reactor was employed to treat saline sulfate wastewater. Mesophilic operation (35 ± 0.5 °C) was performed with hydraulic retention time fixed at 16 h. When the salinity was 28 g L^?1, the chemical oxygen demand and sulfate removal efficiencies were 52 and 67 %, respectively. The salinity effect on sulfate removal was less than that on organics removal. The methane productions were 887 and 329 cm³ L^?1 corresponding to the NaCl concentrations of 12 and 28 g L^?1, respectively. High salinity could stimulate microbes to produce more extracellular polymeric substances (EPSs) and granulation could be performed better. Besides, with the high saline surroundings, a great deal of Na⁺ compressed the colloidal electrical double-layer, neutralized the negative charge of the sludge particles and decreased their electrostatic repulsion. The repulsion barrier disappeared and coagulation took place. The maximum size of granules was 5 mm, which resulted from the coupled triggering forces of high EPSs and Na⁺ contents. Sulfate-reducing bacteria (SRB) were dominant in the high saline surroundings while the methane-producing archaea dominated in the low saline surroundings. The SRB were affected least by the salinity.     

The effect of heavy metals on microbial community structure of a sulfidogenic consortium in anaerobic semi-continuous stirred tank reactors

The effect of heavy metals on community structure of a heavy metal tolerant sulfidogenic consortium was evaluated by using a combination of denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene and dissimilatory sulfite reductase (dsrB) gene fragments, 16S rRNA gene cloning analysis and fluorescence in situ hybridization (FISH). For this purpose, four anaerobic semi-continuous stirred tank reactors (referred as R1–R4) were run in parallel for 12 weeks at heavy metal loading rates of 1.5, 3, 4.5 and 7.5 mg l^?1 d^?1 each of Cu²⁺, Ni²⁺, Zn²⁺, and Cr⁶⁺, respectively. The abundance ratio of Desulfovibrio vulgaris detected by FISH to total cell counts was consistent with the obtained results of cloning and DGGE. This indicated that D. vulgaris was dominant in all analyzed samples and played a key role in heavy metal removal in R1, R2, and R3. In contrast, after 4 weeks of operation of R4, a distinct biomass loss was observed and no positive hybridized cells were detected by specific probes for the domain Bacteria, sulfate-reducing bacteria and D. vulgris. High removal efficiencies of heavy metals were achieved in R1, R2 and R3 after 12 weeks, whereas the precipitation of heavy metals in R4 was significantly decreased after 4 weeks and almost not observed after 6 weeks of operation. In addition, the anaerobic bacteria, such as Pertrimonas sulfuriphila, Clostridium sp., Citrobacter amalonaticus, and Klebsiella sp., identified from DGGE bands and clone library were hypothesized as heavy metal resistant bacteria at a loading rate of 1.5 mg l^?1 d^?1 of Cu²⁺, Ni²⁺, Zn²⁺, and Cr^6+.     

Propionate and butyrate dependent bacterial sulfate reduction at extremely haloalkaline conditions and description of Desulfobotulus alkaliphilus sp. nov.

Evidence on the utilization of simple fatty acids by sulfate-reducing bacteria (SRB) at extremely haloalkaline conditions are practically absent, except for a single case of syntrophy by Desulfonatronum on acetate. Our experiments with sediments from soda lakes of Kulunda Steppe (Altai, Russia) showed sulfide production with sulfate as electron acceptor and propionate and butyrate (but not acetate) as an electron donor at a pH 10–10.5 and a salinity 70–180 g l^?1. With propionate as substrate, a highly enriched sulfidogenic culture was obtained in which the main component was identified as a novel representative of the family Syntrophobacteraceae. With butyrate as substrate, a pure SRB culture was isolated which oxidized butyrate and some higher fatty acids incompletely to acetate. The strain represents the first haloalkaliphilic representative of the family Desulfobacteraceae and is described as Desulfobotulus alkaliphilus sp. nov.     

Kinetics study of pyridine biodegradation by a novel bacterial strain,Rhizobium sp. NJUST18

Biodegradation of pyridine by a novel bacterial strain, Rhizobium sp. NJUST18, was studied in batch experiments over a wide concentration range (from 100 to 1,000 mg l^?1). Pyridine inhibited both growth of Rhizobium sp. NJUST18 and biodegradation of pyridine. The Haldane model could be fitted to the growth kinetics data well with the kinetic constants μ* = 0.1473 h^?1, K _s = 793.97 mg l^?1, K _i = 268.60 mg l^?1 and S _m = 461.80 mg l^?1. The true μ _max, calculated from μ*, was found to be 0.0332 h^?1. Yield coefficient Y _X/S depended on S _i and reached a maximum of 0.51 g g^?1 at S _i of 600 mg l^?1. V _max was calculated by fitting the pyridine consumption data with the Gompertz model. V _max increased with initial pyridine concentration up to 14.809 mg l^?1 h^?1. The q _S values, calculated from $V_{ \hbox{max} }$ , were fitted with the Haldane equation, yielding q _Smax = 0.1212 g g^?1 h^?1 and q* = 0.3874 g g^?1 h^?1 at S _m′ = 507.83 mg l^?1, K _s′ = 558.03 mg l^?1, and K _i′ = 462.15 mg l^?1. Inhibition constants for growth and degradation rate value were in the same range. Compared with other pyridine degraders, μ _max and S _m obtained for Rhizobium sp. NJUST18 were relatively high. High K _i and K _i′ values and extremely high K _s and K _s′ values indicated that NJUST18 was able to grow on pyridine within a wide concentration range, especially at relatively high concentrations.     

In vitro propagation and micromorphological studies of Cleome gynandra: a C4 model plant closely related to Arabidopsis thaliana

We developed a micropropagation protocol for Cleome gynandra, a C₄ model plant with medicinal importance. Surface-sterilized nodal segments obtained from 1 to 2-month-old field grown plant were used as explants for culture establishment and plant regeneration. Multiple shoots differentiated through bud breaking on Murashige and Skoog (MS) medium with different concentrations of benzyladenine (BA) and kinetin (Kin). The optimum shoot differentiation occurred on medium with 1.5 mg l^?1 BA. Out of various concentrations and combinations of cytokinins and auxins, MS medium containing 0.5 mg l^?1 BA and 0.1 mg l^?1 IAA (indole-3-acetic acid) was found best for shoot multiplication. However, the differentiated shoots exhibited hyperhydration, leaf curling and early leaf fall during subculturing. To overcome these problems, regenerated shoots were transferred to the modified MS medium with reduced nitrates (825 mg l^?1 NH₄NO₃ and 950 mg l^?1 KNO₃) and 100 mg l^?1 (NH₄)₂SO₄. The micropropagated shoots were rooted (i) in vitro on one-fourth strength of MS salts with 0.25 mg l^?1 each of IBA (indole-3 butyric acid) and NOA (2-naphthoxyacetic acid) + 100 mg l^?1 activated charcoal, and (ii) ex vitro, by treating the shoot base(s) with 200 mg l^?1 of IBA for 3 min and transferred to soilrite moistened with one-fourth strength of MS macro salts in culture bottles. The plants were hardened in the greenhouse with 85 % survival rate. Micromorphological studies of the plants were conducted during hardening with reference to development and changes in vein spacing, glandular trichome and stomata. In comparison to leaves under in vitro condition, higher density of veins and glandular trichomes was observed in the leaves of hardened plants. In addition, stomata became functional during hardening which were non-functional under in vitro condition.     

Evaluation of 4-bromophenol biodegradation in mixed pollutants system by Arthrobacter chlorophenolicus A6 in an upflow packed bed reactor

Bromophenol is listed as priority pollutant by U.S. EPA, however, there is no report so far on its removal in mixed pollutants system by any biological reactor operated in continuous mode. Furthermore, bromophenol along with chlorophenol and nitrophenol are usually the major constituents of paper pulp and pesticide industrial effluent. The present study investigated simultaneous biodegradation of these three pollutants with specially emphasis on substrate competition and crossed inhibition by Arthrobacter chlorophenolicus A6 in an upflow packed bed reactor (UPBR). A 2³ full factorial design was employed with these pollutants at two different levels by varying their influent concentration in the range of 250–450 mg l^?1. Almost complete removal of all these pollutants and 97 % effluent toxicity removal were achieved in the UPBR at a pollutant loading rate of 1707 mg l^?1 day^?1 or lesser. However, at higher loading rates, the reactor performance deteriorated due to transient accumulation of toxic intermediates. Statistical analysis of the results revealed a strong negative interaction of 4-CP on 4-NP biodegradation. On the other hand, interaction effect between 4-CP and 4-BP was found to be insignificant. Among these three pollutants 4-NP preferentially degraded, however, 4-CP exerted more inhibitory effect on 4-NP biodegradation. This study demonstrated the potential of A. chlorophenolicus A6 for biodegradation of 4-BP in mixed pollutants system by a flow through UPBR system.     

Recovery of blood gases and haematological parameters upon anaesthesia with benzocaine,MS-222 or Aqui-S in the air-breathing catfish <Emphasis Type="Italic">Pangasianodon hypophthalmus</Emphasis>

Fish anaesthesia is used to minimize handling stress and damage during harvesting, transportation, and surgical procedures. Through depression of cardiovascular and respiratory functions, it causes significant changes in blood gases and pH. Here, we present the effects of benzocaine (100 mg l^?1), MS-222 (100 mg l^?1), and Aqui-S (30 mg l^?1) on blood gases and haematological parameters of commercial-sized (≈1 kg) striped catfish (Pangasianodon hypophthalmus) and the time course of recovery. Blood was taken through a dorsal aorta catheter immediately after catheterization, and regularly during the following 72 h recovery in aerated water. All anaesthetics caused increases in PCO₂ and lactate resulting in a decrease in pH_e, closely mirrored by RBC pH_i, as well as a marked rise in Hct, associated with elevated [cortisol] and [glucose] and increased RBC counts but no change in RBC volume, as confirmed by the lack of an adrenergic response of RBC in vitro. All anaesthetics showed similar efficacy and blood parameters were normalized within 24 to 48 h.     

Aerobic decolorization of Acid Brilliant Scarlet GR by microbial community and the community dynamics during sequencing batch processes

In this research, aerobic decolorization of Acid Brilliant Scarlet GR by microbial community was studied. Effects of conditions and dye concentraion on decolorization processes were investigated. Additionally, continuous decolorization was evaluated through sequencing batch tests and the microbial dynamics during this process was analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis. The results showed that 100 mg l^?1 of the dye was completely decolorized within 12 h, which was mainly caused by biodegradation. The optimal decolorization conditions were as follows: inoculation size 2.07 g l^?1 (wet cell pellet), rotation speed 150 r min^?1, pH 5.0–7.0 and 30 °C. The processes were well described by zero-order kinetics, and more than 700 mg l^?1 of the dye would inhibit the activity of the consortium. Furthermore, the microbial community exhibited high efficiency in sequencing batch processes for continuous decolorization. Microbial community structure shifted obviously when exposed to higher concentration of the dye (500 mg l^?1), and all the dominant microorganisms were affiliated with four different phyla of Actinobacteria, Bacteroidetes, Proteobacteria and Firmicutes.     

Agrobacterium-mediated transformation of Eruca sativa

An Agrobacterium tumefaciens—mediated transformation system was developed for Eruca sativa (eruca). Hypocotyl explants were co-cultivated with bacterial cells carrying a plasmid harboring a uidA:nptII fusion gene along a phosphinothricin acetyl transferase (PAT) gene cassette, for a period of 2 days. These were grown on a high cytokinin/auxin medium containing 5.0 mg l^?1 6-benzyladenine (BA), 1.0 mg l^?1 indole-3-acetic acid (IAA), and 0.1 mg l^?1 α-naphthaleneacetic acid (NAA). Explants were then transferred to a lower cytokinin/auxin medium containing 2.0 mg l^?1 BA and 0.1 mg l^?1 NAA along with 5.0 mg l^?1 silver nitrate and 300 mg l^?1 Timentin®. Upon transfer to a selection medium containing either 20 mg l^?1 kanamycin or 2 mg l^?1 L-phosphinothricin (L-ppt), shoot regenerants were observed. Expression of the transgenes in putative transformants was confirmed using a histochemical GUS assay. Presence of the PAT transgene in GUS-positive T0 plants was confirmed by Southern blot analysis. Moreover, spot tests of T1 seedlings were conducted using the L-ppt herbicide. A transformation frequency of 1.1% was obtained with more than 60% of transgenic lines containing single copies of the transgenes.     

Plant regeneration via callus culture and subsequent in vitro flowering of Dendrobium huoshanense

A protocol for regenerating and subsequent in vitro flowering of an economical important and endangered medicinal orchid, Dendrobium huoshanense, was established mainly via indirect protocorm-like body (PLB) formation. A four-step method was developed to induce successful plant regeneration on 1/2 MS medium supplemented with suitable plant growth regulators (PGRs). Step 1 (callus induction): the root tip explants (1 cm long) were cultured at 1 mg l^?1 2,4-D + 1 mg l^?1 TDZ for 3 months. Step 2 (callus proliferation): the calli were subcultured with a 1-month interval at 1 mg l^?1 2,4-D + 1 mg l^?1 TDZ. Step 3 (PLB induction): the calli were cultured at 2 mg l^?1 NAA + 1 mg l^?1 BA for 2 months. Step 4 (plantlet conversion): the 2-month-old PLBs were cultured at 0.1 mg l^?1 IBA for 4 months. It took at least 6 months to produce well-rooted regenerated plantlets with an average of 3.2 roots and 3.6 leaves from the initial callus. The 6-month-old rooted plantlets were transferred onto PGR-free 1/2 MS medium for 6 months, and then potted with Sphagnum moss for acclimatization. After 2 month of culture, the survival rate was 100 %. The in vitro flowers were obtained on the 8-month-old plantlets at 1 mg l^?1 IBA, 5 mg l^?1 IBA and 0.1 mg l^?1 NAA, but the flowers showed a lack of the gynandrium. The abnormity was overcome by the aid of 5 mg l^?1 TDZ, and subsequently, the capsules formed without artificial pollination. This protocol provides the basis for further investigation on cell suspension, micropropagation, in vitro flowering and breeding programs in Dendrobium huoshanense.     

Efficacious somatic embryogenesis and fertile plant recovery from shoot apex explants of onion (Allium cepa. L.)

An efficient somatic embryogenesis and regeneration system was developed for the first time in onion using shoot apex explants. These explants were used to initiate callus in Murashige and Skoog (MS) medium supplemented with 4.0 mg l^?1 2,4-dichlorophenoxyacetic acid. The induction frequency of primary callus in this medium was 85.3%. The primary calli were then transferred onto medium supplemented with 2.0 mg l^?1 2,4-dichlorophenoxyacetic acid. Following two biweekly subcultures, embryogenic callus formed. Inclusion of a low concentration of 6-benzylaminopurine in the subculture medium promoted the formation of embryogenic callus. The addition of 2.0 mg l^?1 glycine, 690 mg l^?1 proline, and 1.0 g l^?1 casein hydrolysate also increased the frequency of callus induction and embryogenic callus formation. The highest frequency of embryogenic callus (86.9%) and greatest number of somatic embryos (26.3 per callus) were obtained by the further addition of 8.0 mg l^?1 silver nitrate. Somatic embryos formed plantlets on regeneration medium supplemented with 1.5 mg l^?1 6-benzylaminopurine; addition of 2.0 mg l^?1 glycine to the regeneration medium promoted a high frequency of regeneration (78.1%) and plantlet formation (28.7 plants per callus). The regenerated plantlets were transferred to half-strength MS medium supplemented with 1.5 mg l^?1 indole-3-butyric acid for root development; the maximum frequency of root formation was 87.7% and the average number of roots was 7.6 per shoot. The regenerated plantlets were successfully grown to maturity after hardening in the soil. This is the first report of somatic embryogenesis and regeneration from shoot apex explants of onion.     

Production of aglycone protopanaxatriol from ginseng root extract using Dictyoglomus turgidum β-glycosidase that specifically hydrolyzes the xylose at the C-6 position and the glucose in protopanaxatriol-type ginsenosides

The hydrolytic activity of a recombinant β-glycosidase from Dictyoglomus turgidum that specifically hydrolyzed the xylose at the C-6 position and the glucose in protopanaxatriol (PPT)-type ginsenosides followed the order Rf > Rg₁ > Re > R₁ > Rh₁ > R₂. The production of aglycone protopanaxatriol (APPT) from ginsenoside Rf was optimal at pH 6.0, 80 °C, 1 mg ml^?1 Rf, and 10.6 U ml^?1 enzyme. Under these conditions, D. turgidum β-glycosidase converted ginsenoside R₁ to APPT with a molar conversion yield of 75.6 % and a productivity of 15 mg l^?1 h^?1 after 24 h by the transformation pathway of R₁ → R₂ → Rh₁ → APPT, whereas the complete conversion of ginsenosides Rf and Rg₁ to APPT was achieved with a productivity of 1,515 mg l^?1 h^?1 after 6.6 h by the pathways of Rf → Rh₁ → APPT and Rg₁ → Rh₁ → APPT, respectively. In addition, D. turgidum β-glycosidase produced 0.54 mg ml^?1 APPT from 2.29 mg ml^?1 PPT-type ginsenosides of Panax ginseng root extract after 24 h, with a molar conversion yield of 43.2 % and a productivity of 23 mg l^?1 h^?1, and 0.62 mg ml^?1 APPT from 1.35 mg ml^?1 PPT-type ginsenosides of Panax notoginseng root extract after 20 h, with a molar conversion yield of 81.2 % and a productivity of 31 mg l^?1 h^?1. This is the first report on the APPT production from ginseng root extract. Moreover, the concentrations, yields, and productivities of APPT achieved in the present study are the highest reported to date.     

Effect of the cathode potential and sulfate ions on nitrate reduction in a microbial electrochemical denitrification system

Recently, bioelectrochemical systems have been demonstrated as advantageous for denitrification. Here, we investigated the nitrate reduction rate and bacterial community on cathodes at different cathode potentials [?300, ?500, ?700, and ?900 mV vs. standard hydrogen electrode (SHE)] in a two-chamber microbial electrochemical denitrification system and effects of sulfate, a common nitrate co-contaminant, on denitrification efficiency. The results indicated that the highest nitrate reduction rates (3.5 mg L^?1 days^?1) were obtained at a cathode potential of ?700 mV, regardless of sulfate presence, while a lower rate was observed at a more negative cathode potential (?900 mV). Notably, although sulfate ions generally inhibited nitrate reduction, this effect was absent at a cathode potential of ?700 mV. Polymerase chain reaction–denaturing gradient gel electrophoresis revealed that bacterial communities on the graphite-felt cathode were significantly affected by the cathode potential change and sulfate presence. Shinella-like and Alicycliphilus-like bacterial species were exclusively observed on cathodes in reactors without sulfate. Ochrobactrum-like and Sinorhizobium-like bacterial species, which persisted at different cathode potentials irrespective of sulfate presence, were shown to contribute to bioelectrochemical denitrification. This study suggested that a cathode potential of around ?700 mV versus SHE would ensure optimal nitrate reduction rate and counteract inhibitory effects of sulfate. Additionally, sulfate presence considerably affects denitrification efficiency and microbial community of microbial electrochemical denitrification systems.     

Optimizing micropropagation of drought resistant <Emphasis Type="Italic">Pyrus boissieriana</Emphasis> Buhse

The present study concentrated on introducing a micropropagation protocol for a drought resistant genotype from Pyrus boissieriana, which is the second most naturally widespread pear species in Iran with proper physiological and medicinal properties. Proliferating microshoot cultures were obtained by placing nodal segments on MS medium supplemented with BAP and IBA or NAA. The highest number of shoots (27 shoots per explant) were obtained with 1.5 mg l^?1 BAP and 0.05 mg l^?1 IBA, but this combination did not produce shoots of desirable length (>1.7 cm). Combination of 1.75 mg l^?1 BAP and 0.07 mg l^?1 IBA was the best for the shoot multiplication in P. boissieriana with a sufficient number of shoot production (22.33 shoots per explant) and relatively more appropriate shoot length. The larger and greenish leaves were obtained when PG was added to the best multiplication treatment. Microshoot elongation was carried out in 1/2 and 1/4 MS medium containing 50–100 mg l^?1 PG with different concentrations of IBA or NAA at intervals of 30–60 days. Significant increase in shoot length was detected after 45–60 days of culture in the presence of PG. The highest shoot length (8 cm) was recorded on 1/2 MS medium supplemented with 0.5 mg l^?1 IBA and 100 mg l^?1 PG. GA₃ negatively affected number and length of shoots and generally caused generation of red leaves. The highest percentage of root induction (100%) and root length (9 cm) were obtained on 1/6 strength MS medium supplemented with 0.005 mg l^?1 IBA. All plantlets were hardened when transferred to ex vitro conditions through a period of 25–30 days. The results suggest axillary shoot proliferation of P. boissieriana could successfully be employed for propagation of candidate drought resistant seedling.     

Potential availability of anaerobic treatment with digester slurry of animal waste for the reclamation of acid mine water containing sulfate and heavy metals

The use of an anaerobic digester slurry of cattle waste for the reclamation of acid mine water was examined. When the digester slurry was mixed with acid mine water, anaerobic digestion, including sulfate reduction and methanogenesis, was enhanced. In the mixture of acid mine water and the digester slurry, sulfate reduction proceeded without diminishing methanogenesis. The digester slurry and its supernatant (SDF-sup) showed a significant capacity to act as a strong alkaline reagent, and the pH of the acid mine water was markedly elevated by the addition of the digester slurry of SDF-sup even at the low ratio of 1% (v/v). Precipitation of heavy metals in the acid mine water occurred as the pH was elevated by the addition of SDF-sup. When the digester slurry was added at the ratio of 5% (v/v) to acid mine water which had been pretreated with SDF-sup, the rate of sulfate reduction increased with increasing the concentration of sulfate in the mixture up to about 1,400 mg·l⁻¹. In acid mine water pretreated with SDF-sup and supplemented with the digester slurry at the ratio of 5% (v/v), the maximum amount of sulfate reduced within 20 d of incubation was about 1,000 mg·l⁻¹, and the maximum rate of sulfate reduction was about 120 mg SO₄²⁻·l⁻¹·d⁻¹.