Competition for electrons between mono-oxygenations of pyridine and 2-hydroxypyridine

Pyridine and its heterocyclic derivatives are widely encountered in industrial wastewaters, and they are relatively recalcitrant to biodegradation. Pyridine biodegradation is initiated by two mono-oxygenation reactions that compete for intracellular electron donor (2H). In our experiments, UV photolysis of pyridine generated succinate, whose oxidation augmented the intracellular electron donor and accelerated pyridine biodegradation and mineralization. The first mono-oxygenation reaction always was faster than the second one, because electrons provided by intracellular electron donors were preferentially utilized by the first mono-oxygenase; this was true even when the concentration of 2HP was greater than the concentration of pyridine. In addition, the first mono-oxygenation had faster kinetics because it had higher affinity for its substrate (pyridine), along with less substrate self-inhibition.     

UV photolysis for accelerated quinoline biodegradation and mineralization

Sequentially and intimately coupled photolysis with biodegradation were evaluated for their ability to accelerate quinoline-removal and quinoline-mineralization kinetics. UV photolysis sequentially coupled to biodegradation significantly improved biomass-growth kinetics, which could be represented well by the Aiba self-inhibition model: UV photolysis increased the maximum specific growth rate (μ _max) by 15 %, and the inhibition constant (K _SI) doubled. An internal loop photo-biodegradation reactor (ILPBR) was used to realize intimately coupled photolysis with biodegradation. The ILPBR was operated with batch experiments following three protocols: photolysis alone (P), biodegradation alone (B), and intimately coupled photolysis and biodegradation (P&B). For P&B, the maximum quinoline removal rate (r _max) increased by 9 %, K _SI increased by 17 %, and the half-maximum-rate concentration (K _S) decreased by 55 %, compared to B; the composite result was a doubling of the quinoline-biodegradation rate for most of the concentration range tested. The degree of mineralization was increased by both forms of photolysis coupled to biodegradation, and the impact was greater for intimate coupling (18 % increase) than sequential coupling (5 %). The benefits of UV photolysis were greater with intimate coupling than with sequential coupling due to parallel transformation by biodegradation and photolysis.     

UV photolysis for enhanced phenol biodegradation in the presence of 2,4,6-trichlorophenol (TCP)

A bacterial strain isolated from activated sludge and identified as Bacillus amyloliquefaciens could biodegrade phenol, but 2,4,6-trichlorophenol (TCP) inhibited phenol biodegradation and biomass growth. UV photolysis converted TCP into dichlorocatechol, monochlorophenol, and dichlorophenol, and this relieved inhibition by TCP. Phenol-removal and biomass-growth rates were significantly accelerated after UV photolysis: the monod maximum specific growth rate (μ _max) increased by 9 % after TCP photolysis, and the half-maximum-rate concentration (K _S) decreased by 36 %. Thus, the major benefit of UV photolysis in this case was to transform TCP into a set of much-less-inhibitory products.     

2,4-DNT removal in intimately coupled photobiocatalysis: the roles of adsorption, photolysis, photocatalysis, and biotransformation

The removal of 2,4-dinitrotoluene (2,4-DNT) by simultaneous UV-photo(cata)lysis and biodegradation was explored using intimately coupled photolysis/photocatalysis and biodegradation (ICPB) with two novel porous carriers. First, a porous ceramic carrier was used to attach the photocatalyst (TiO?) on its exterior and accumulate biomass in its interior. UV irradiation alone decomposed 71% of the 2,4-DNT in 60 h, and TiO? catalyst improved the photolysis to 77%. Second, a macroporous sponge carrier was used to strongly adsorb 2,4-DNT and protect microorganisms from 2,4-DNT inhibition and UV irradiation. The main photolytic reactions were reduction of the nitryl to amino and hydrolysis of the amino to release NH??. The main biodegradation reactions were oxidative release of NO?? and accelerated reductive release of NH??. ICPB more thoroughly released inorganic N, with nearly equal amounts being oxidized to nitrate and reduced to ammonium. The genera Burkholderia and Bacillus were found inside the sponge carriers, and they are associated with biodegradation of DNT and its photolysis intermediates. Therefore, using an adsorbent and macroporous biofilm carrier enabled the effective removal of 2,4-DNT by ICPB.     

The role of exogenous electron donors for accelerating 2,4,6-trichlorophenol biotransformation and mineralization

2,4,6-Trichlorophenol (TCP) is a biologically recalcitrant compound, but its biodegradation via reductive dechlorination can be accelerated by adding an exogenous electron donor. In this work, acetate and formate were evaluated for their ability to accelerate TCP reductive dechlorination, as well to accelerate mono-oxygenation of TCP’s reduction product, phenol. Acetate and formate accelerated TCP reductive dechlorination, and the impact was proportional to the number of electron equivalents released by oxidation of the donor: 8 e^? equivalents per mol for acetate, compared to 2 e^? eq per mol for formate. The acceleration phenomenon was similar for phenol mono-oxygenation, and this increased the rate of TCP mineralization. Compared to endogenous electron equivalents generated by phenol mineralization, the impact of exogenous electron donor was stronger on a per-equivalent basis.     

Integrated photocatalytic-biological reactor for accelerated 2,4,6-trichlorophenol degradation and mineralization

An integrated photocatalytic-biological reactor (IPBR) was used for accelerated degradation and mineralization of 2,4,6-trichlorophenol (TCP) through simultaneous, intimate coupling of photocatalysis and biodegradation in one reactor. Intimate coupling was realized by circulating the IPBR’s liquid contents between a TiO₂ film on mat glass illuminated by UV light and honeycomb ceramics as biofilm carriers. Three protocols—photocatalysis alone (P), biodegradation alone (B), and integrated photocatalysis and biodegradation (photobiodegradation, P&B)—were used for degradation of different initial TCP concentrations. Intimately coupled P&B also was compared with sequential P and B. TCP removal by intimately coupled P&B was faster than that by P and B alone or sequentially coupled P and B. Because photocatalysis relieved TCP inhibition to biodegradation by decreasing its concentration, TCP biodegradation could become more important over the full batch P&B experiments. When phenol, an easy biodegradable compounds, was added to TCP in order to promote TCP mineralization by means of secondary utilization, P&B was superior to P and B in terms of mineralization of TCP, giving 95% removal of chemical oxygen demand. Cl⁻ was only partially released during P experiments (24%), and this corresponded to its poor mineralization in P experiments (32%). Thus, intimately coupled P&B in the IPBR made it possible obtain the best features of each: rapid photocatalytic transformation in parallel with mineralization of photocatalytic products.     

Sequential degradation of <Emphasis Type="Italic">p</Emphasis>-cresol by photochemical and biological methods

Sequential photo-and biodegradation of p-cresol was studied using a mercury lamp, as well as KrCl and XeCl excilamps. Preirradiation of p-cresol at a concentration of 10^?4 M did not affect the rate of its subsequent biodegradation. An increase in the concentration of p-cresol to 10^?3 M and in the duration preliminary UV irradiation inhibited subsequent biodegradation. Biodegradation of p-cresol was accompanied by the formation of a product with a fluorescence maximum at 365 nm (λ_ex = 280 nm), and photodegradation yielded a compound fluorescing at 400 nm (λ_ex = 330 nm). Sequential UV and biodegradation led to the appearance of bands in the fluorescence spectra that were ascribed to p-cresol and its photolysis products. It was shown that sequential use of biological and photochemical degradation results in degradation of not only the initial toxicant but also the metabolites formed during its biodegradation.     

Integrated photocatalytic-biological reactor for accelerated phenol mineralization

An integrated photocatalytic-biological reactor (IPBR) was developed for accelerated phenol degradation and mineralization. In the IPBR, photodegradation and biodegradation occurred simultaneously, but in two separated zones: a piece of mat-glass plate coated with TiO₂ film and illuminated by UV light was connected by internal circulation to a honeycomb ceramic that was the biofilm carrier for biodegradation. This arrangement was designed to give intimate coupling of photocatalysis and biodegradation. Phenol degradation was investigated by following three protocols: photocatlysis with TiO₂ film under ultraviolet light, but no biofilm (photodegradation); biofilm biodegradation with no UV light (biodegradation); and simultaneous photodegradation and biodegradation (intimately coupled photobiodegradation). Photodegradation alone could partly degrade phenol, but was not able to achieve significant mineralization, even with an HRT of 10 h. Biodegradation alone could completely degrade phenol, but it did not mineralize the COD by more than 74%. Photobiodegradation allowed continuous rapid degradation of phenol, but it also led to more complete mineralization of phenol (up to 92%) than the other protocols. The results demonstrate that intimate coupling was achieved by protecting the biofilm from UV and free-radical inhibition. With phenol as the target compound, the main advantage of intimate coupling in the IPBR was increased mineralization, presumably because photocatalysis made soluble microbial products more rapidly biodegradable.     

Effects of inorganic nanoparticles on viability and catabolic activities of Agrobacterium sp. PH-08 during biodegradation of dibenzofuran

This study investigated the cytotoxicity, genotoxicity, and growth inhibition effects of four different inorganic nanoparticles (NPs) such as aluminum (nAl), iron (nFe), nickel (nNi), and zinc (nZn) on a dibenzofuran (DF) degrading bacterium Agrobacterium sp. PH-08. NP (0–1,000 mg L^?1) -treated bacterial cells were assessed for cytotoxicity, genotoxicity, growth and biodegradation activities at biochemical and molecular levels. In an aqueous system, the bacterial cells treated with nAl, nZn and nNi at 500 mg L^?1 showed significant reduction in cell viability (30–93.6 %, p < 0.05), while nFe had no significant inhibition on bacterial cell viability. In the presence of nAl, nZn and nNi, the cells exhibited elevated levels of reactive oxygen species (ROS), DNA damage and cell death. Furthermore, NP exposure showed significant (p < 0.05) impairment in DF and catechol biodegradation activities. The reduction in DF biodegradation was ranged about 71.7–91.6 % with single NPs treatments while reached up to 96.3 % with a mixture of NPs. Molecular and biochemical investigations also clearly revealed that NP exposure drastically affected the catechol-2,3-dioxygenase activities and its gene (c23o) expression. However, no significant inhibition was observed in nFe treatment. The bacterial extracellular polymeric materials and by-products from DF degradation can be assumed as key factors in diminishing the toxic effects of NPs, especially for nFe. This study clearly demonstrates the impact of single and mixed NPs on the microbial catabolism of xenobiotic-degrading bacteria at biochemical and molecular levels. This is the first study on estimating the impact of mixed NPs on microbial biodegradation.     

Biodegradation of sulfonamides by <Emphasis Type="Italic">Shewanella oneidensis</Emphasis> MR-1 and <Emphasis Type="Italic">Shewanella</Emphasis> sp. strain MR-4

Because of extensive sulfonamides application in aquaculture and animal husbandry and the consequent increase in sulfonamides discharged into the environment, strategies to remediate sulfonamide-contaminated environments are essential. In this study, the resistance of Shewanella oneidensis MR-1 and Shewanella sp. strain MR-4 to the sulfonamides sulfapyridine (SPY) and sulfamethoxazole (SMX) were determined, and sulfonamides degradation by these strains was assessed. Shewanella oneidensis MR-1 and Shewanella sp. strain MR-4 were resistant to SPY and SMX concentrations as high as 60 mg/L. After incubation for 5 days, 23.91 ± 1.80 and 23.43 ± 2.98% of SPY and 59.88 ± 1.23 and 63.89 ± 3.09% of SMX contained in the medium were degraded by S. oneidensis MR-1 and Shewanella sp. strain MR-4, respectively. The effects of the initial concentration of the sulfonamides and initial pH of the medium on biodegradation, and the degradation of different sulfonamides were assessed. The products were measured by LC–MS; with SPY as a substrate, 2-AP (2-aminopyridine) was the main stable metabolite, and with SMX as a substrate, 3A5MI (3-amino-5-methyl-isoxazole) was the main stable metabolite. The co-occurrence of 2-AP or 3A5MI and 4-aminobenzenesulfonic acid suggests that the initial step in the biodegradation of the two sulfonamides is S–N bond cleavage. These results suggest that S. oneidensis MR-1 and Shewanella sp. strain MR-4 are potential bacterial resources for biodegrading sulfonamides and therefore bioremediation of sulfonamide-polluted environments.     

Effects of high glucose and high insulin concentrations on osteoblast function in vitro

Bone disease as a consequence of diabetes mellitus (DM) is not fully understood. The effects of high glucose (30 mM), high insulin (50 nM), or mannitol (30 mM; osmotic control) were evaluated on MC3T3-E1 cells (osteoblasts) in vitro. The mRNA and protein levels of parathyroid hormone (PTH) receptor (PTH1R), collagen I, RANKL, osteoprotegerin (OPG), alkaline phosphatase (ALP), and glucose transporter (GLUT1) were estimated by real-time polymerase chain reaction or Western blotting. The mineralization capacity was analyzed by von Kossa staining. High glucose induced overexpression of RANKL (2×) and OPG (30×), suggesting that RANKL-induced osteoclast activity might not be a dominant mechanism of bone disease in DM, since this increase was followed by increased OPG. Collagen I increased by 12×, indicating an excess of organic matrix production. The expression of ALP decreased by 50 %, indicating a deficit in mineralization capacity, confirmed by von Kossa staining. Mannitol induced similar effects as glucose suggesting that extracellular hyperosmolarity was able to stimulate organic matrix production. GLUT1 expression was not altered, and insulin did not reverse most of the effects of glucose, suggesting that glucose uptake by osteoblasts was not altered by high glucose. The data suggest that the bone fragility typical of DM is not a consequence of excessive bone reabsorption but is instead attributable to a defect in organic matrix mineralization. The heightened increase in OPG versus RANKL might cause a decrease in the bone-remodeling cycle. Osteoblasts appear to be more sensitive to extracellular hypertonicity than to the intracellular metabolic effects of hyperglycemia.     

Sequential degradation of p-cresol by photochemical and biological methods

Sequential photo- and biodegradation of p-cresol was studied using a mercury lamp, as well as KrCl and XeCl excilamps. Preirradiation of p-cresol at a concentration of 10(-4) M did not affect the rate of its subsequent biodegradation. An increase in the concentration of p-cresol to 10(-3) M and in the duration preliminary UV irradiation inhibited subsequent biodegradation. Biodegradation of p-cresol was accompanied by the formation of a product with a fluorescence maximum at 365 nm (lambdaex 280 nm), and photodegradation yielded a compound fluorescing at 400 nm (lambdaex 330 nm). Sequential UV and biodegradation led to the appearance of bands in the fluorescence spectra that were ascribed to p-cresol and its photolysis products. It was shown that sequential use of biological and photochemical degradation results in degradation of not only the initial toxicant but also the metabolites formed during its biodegradation.     

Photobiodegradation of phenol with ultraviolet irradiation of new ceramic biofilm carriers

Activated sludge acclimated to biodegrade phenol was allowed to attach on and in light porous ceramic carriers and to function as a biofilm in a photolytic circulating-bed bioreactor (PCBBR). Phenol degradation in the PCBBR was investigated following three protocols: photolysis with ultraviolet light alone (P), biodegradation alone (B), and the two mechanisms operating simultaneously (P/B). Phenol was degraded at approximately equal rates by B and P/B, each of which was much faster than the rate by P. Furthermore, phenol was mineralized to a significantly greater extent with P/B than with either P or B. SEM showed that the biofilm survived well inside macropores that presumably shaded the microorganisms from UV irradiation, even though the UV light greatly reduced biofilm on outer surface of the carriers in the P/B experiments. Rapid biodegradation of phenol, enhanced mineralization, and survival of bacteria inside macropores demonstrated that being in a biofilm inside the porous carriers protected the bacteria from UV-light toxicity, allowing intimate coupling of photolysis and biodegradation.     

Anaerobic biodegradation of high-molecular-weight polycyclic aromatic hydrocarbons by a facultative anaerobe Pseudomonas sp. JP1

Polycyclic aromatic hydrocarbons (PAHs) are harmful persistent organic pollutants, while the high-molecular-weight (HMW) PAHs are even more detrimental to the environment and human health. However, microbial anaerobic degradation of HMW PAHs has rarely been reported. One facultative anaerobe Pseudomonas sp. JP1 was isolated from Shantou Bay, Shantou, China, which could degrade a variety of HMW PAHs. After 40 days cultivation with strain JP1, anaerobic biodegradation rate of benzo[a]pyrene (BaP), fluoranthene, and phenanthrene was 30, 47, and 5 %, respectively. Consumption of nitrate as the electron acceptor was confirmed by N-(1-naphthyl) ethylenediamine spectrophotometry. Supplementation of sodium sulfite, maltose, or glycine, and in a salinity of 0–20 ‰ significantly stimulated anaerobic degradation of BaP. Lastly, the anaerobic degradation metabolites of BaP by strain JP1 were investigated using GC/MS, and the degradation pathway was proposed. This study is helpful for further studies on the mechanism of anaerobic biodegradation of PAHs.     

Differential Effects of Fluoride During Osteoblasts Mineralization in C57BL/6J and C3H/HeJ Inbred Strains of Mice

The behavior of fluoride ions in biological systems has advantages and problems. On one hand, fluoride could be a mitogenic stimulus for osteoblasts. However, high concentrations of this element can cause apoptosis in rat and mouse osteoblasts. Toward an understanding of this effect, we examined the role of sodium fluoride (NaF) in two mouse calvaria osteoblasts during the mineralization process. The animals used were C3H/HeJ (C3) and C57BL/6J (B6) mice. The calvaria cells were cultured for 28 days in the presence of several doses of NaF (0, 5, 10, 25, 50, and 75 μM), and we performed the assays: mineralized nodule measurements, alkaline phosphatase (ALP) activity, determination of type I collagen, and matrix metalloproteinase-2 (MMP-2) activity. The results showed no effects on alkaline phosphatase activity but decreased mineralized nodule formation. In B6 cells, the NaF effect was already seen with 10 μM of NaF and a greater increase of cellular type I collagen, and MMP-2 activity was upregulated after 7 days of NaF exposure. C3 osteoblasts showed a reduction in the mineralization pattern only after 50 μM of NaF with a slight increase of type I collagen and downregulation of MMP-2 activity during the mineralization period. In conclusion, fluoride affects the production and degradation of the extracellular matrix during early onset and probably during the mineralization period. Additionally, the genetic factors may contribute to the variation in cell response to fluoride exposure, and the differences observed between the two strains could be explained by an alteration of the bone matrix metabolism (synthesis and degradation).     

Internal loop photobiodegradation reactor (ILPBR) for accelerated degradation of sulfamethoxazole (SMX)

The internal loop photobiodegradation reactor (ILPBR) was evaluated for the degradation of the pharmaceutical sulfamethoxazole (SMX) using batch experiments following three protocols: photolysis alone (P), biodegradation alone (B), and intimately coupled photolysis and biodegradation (P&B). SMX was removed more rapidly by P&B than by either P or B alone, and the corresponding dissolved organic carbon (DOC) removals by P&B also were higher. The faster SMX removal probably was due to a synergy between photolysis and the rapid biodegradation of SMX by the biofilm. The greater DOC removal was brought about by the presence of biofilm bacteria able to biodegrade photolysis products. Ammonium N released during photolysis of SMX gave more evidence for the formation of intermediates and was enough in P&B experiments to support bioactivity when no other N was supplied. Clone libraries performed on the biofilms before and after the P&B experiments showed profound changes in the microbial community. Whereas Rhodopirellula baltica and Methylibium petroleiphilum PM1 dominated the biofilm after the B experiments, they were replaced by Micrococcus luteus, Delftia acidovorans, and Oligotropha carboxidovorans after the P&B experiments. The changes in microbial community structure mirrored the change in function in the P&B experiments: SMX biodegradation (presumably the roles of R. baltica and M. petroleiphilum) was out-competed by SMX photolysis, but biodegradation of photolysis products (most likely by M. luteus and D. acidovorans) became important. The higher removal rates of SMX and DOC, as well as the changes in microbial community structure, confirm the value of intimately coupling photolysis with biodegradation in the ILPBR.     

Which way is up? Asymmetric spectral input along the dorsal–ventral axis influences postural responses in an amphibious annelid

Medicinal leeches are predatory annelids that exhibit countershading and reside in aquatic environments where light levels might be variable. They also leave the water and must contend with terrestrial environments. Yet, leeches generally maintain a dorsal upward position despite lacking statocysts. Leeches respond visually to both green and near-ultraviolet (UV) light. I used LEDs to test the hypothesis that ventral, but not dorsal UV would evoke compensatory movements to orient the body. Untethered leeches were tested using LEDs emitting at red (632 nm), green (513 nm), blue (455 nm) and UV (372 nm). UV light evoked responses in 100 % of trials and the leeches often rotated the ventral surface away from it. Visible light evoked no or modest responses (12–15 % of trials) and no body rotation. Electrophysiological recordings showed that ventral sensilla responded best to UV, dorsal sensilla to green. Additionally, a higher order interneuron that is engaged in a variety of parallel networks responded vigorously to UV presented ventrally, and both the visible and UV responses exhibited pronounced light adaptation. These results strongly support the suggestion that a dorsal light reflex in the leech uses spectral comparisons across the dorsal–ventral axis rather than, or in addition to, luminance.     

Photochemistry of NH3, CH4 and PH3. Possible applications to the Jovian planets

Photolysis of NH₃ at 185 nm in the presence of a two-fold excess of CH₄ results in the loss of about 0.25 mole of CH₄ per mole of NH₃ decomposed (ΔCH₄/ΔNH₃). The loss arises from the abstraction of hydrogen atoms from CH₄ by photolytically generated hot hydrogen atoms, the presence of which is established by the constancy of ΔCH₄/ΔNH₃ between 298 and 156 K and by the quenching of the abstraction reaction when either H₂ or SF₆ is added. From the latter result, it can be concluded that NH₃ photolysis in the H₂-abundant atmosphere of Jupiter is not responsible for the presence of the carbon compounds observed there such as ethane, acetylene, and hydrogen cyanide, but may have had a role in the early atmosphere of Titan. Photolysis of PH₃ with a 206 nm light source gives P₂H₄, which in turn is converted to a red-brown solid (P₄?). The course of the photolysis is not changed appreciably when the temperature is lowered to 157 K except that the concentration of P₂H₄ increases. The presence of H₂ has no effect on the P₂H₄ yield. Photolysis of 9∶1 NH₃∶PH₃ gives a rate of decomposition of PH₃ that is comparable with that observed by the direct photolysis of PH₃. Comparable amounts of P₂H₄ and the red-brown solid are also observed. The mechanisms of these photochemical reactions together with their implications to the atmospheric chemistry of Jupiter are discussed. The structures of the compounds responsible for the wide array of colorse.g., brown, red and white, observed in the atmosphere of Jupiter have been the subject of extensive speculation. One theory suggests that these colors are due to organic materials formed by the action of either solar ultraviolet light or electric discharges on mixtures of CH₄, NH₃ and NH₄HS in the Jovian atmosphere (Ponnamperuma, 1976; Khareet al., 1978). An alternative hypothesis is that the colors are due to inorganic compounds resulting from the photolysis of NH₄HS and PH₃ (Lewis and Prinn, 1970; Prinn and Lewis, 1975). In this paper we will summarize our experiments which were designed to test some of these hypotheses.