Anaerobic degradation of dehydrodiisoeugenol by rumen bacteria

The degradation of dehydrodiisoeugenol (DDIE) by cow rumen bacteria was studied under strictly anaerobic conditions. After two days of cultivation, about 23% of DDIE (1.2 mM) was degraded to volatile fatty acids (VFA) such as acetic acid, propionic acid and butyric acid. The aromatic intermediates were vanillic acid, 5-methylvanillin and 3-methyl-4-hydroxybenzaldehyde, which suggested that the coumaran ring in DDIE was cleaved during degradation. These results indicate that the rumen anaerobes can degrade this lignin-related dimer to monoaromatic compounds and VFA.     

O-Demethylation, dehydroxylation, ring-reduction and cleavage of aromatic substrates by Enterobacteriaceae under anaerobic conditions

Four fermentative facultative anaerobes, members of the genera Enterobacter and Escherichia , were tested for their ability to transform an aromatic lignin derivative, 3-methoxy-4-hydroxy-cinnamic acid (ferulic acid), under anaerobic (fermentative) conditions. The pure cultures studied were shown to O-demethylate, dehydroxylate, reduce the double bond in the side-chain, decarboxylate the aromatic ring to the stage of benzoate and to reduce the ring to an alicyclic acid. Aromatic hydrocarbons (toluene, ethylbenzene and propylbenzene), as well as phenols (phenol, o-cresol, p-cresol, 2-ethylphenol and 3-hydroxy-4-ethylphenol) were also produced. In addition, during 3 months incubation, the cleavage of the aromatic ring occurred, whereby a small fraction of the substrate was converted to straight-chain and branched (methylated, ethylated) five- to eight-carbon aliphatic acids. The results indicate that pure cultures of fermentative facultative anaerobes might be capable of degrading substituted aromatic acids to aliphatic products under strictly anaerobic (fermentative) conditions. These abilities, which have so far been found only in denitrifying pseudomonads among facultative anaerobes, might be common in Enterobacteriaceae. It is conceivable that these bacteria are important as degraders of aromatic compounds in anaerobic ecosystems.     

Characteristics of bacteria isolated by the anaerobic roll-tube method from cheeses and ground beef.

In this study the methods of Hungate were used to quantitate the anaerobic bacteria present in commercially available ground beef, cheddar cheese, and German hand cheese. Of 235 anaerobic roll-tube isolates from ground beef and German hand cheese, all were facultative anaerobes. Of 213 anaerobic roll-tube isolates from cheddar cheese, 91% were facultative anaerobes and 9% were obligate anaerobes. Using results of biochemical tests, 14 or the 17 obligately anaerobic isolates from cheddar cheese were Propionibacterium acnes, two were strains of Propionibacterium that could not be speciated, and one was tentatively identified as a strain of Streptococcus evolutus. Obligate anaerobes were estimated to be present in the cheddar cheese at a level of about 10(6)/g. The possible significance of these levels of P. acnes in nonsterile foods is discussed.     

Anaerobic digestion of swine manure under natural zeolite addition: VFA evolution, cation variation, and related microbial diversity

Batch experiments were carried out on anaerobic digestion of swine manure under 10 % of total solids and 60 g/L of zeolite addition at 35 °C. Four distinctive volatile fatty acid (VFAs) evolution stages were observed during the anaerobic process, i.e., VFA accumulation, acetic acid (HAc) and butyric acid (HBu) utilization, propionic acid (HPr) and valeric acid (HVa) degradation, and VFA depletion. Large decreases in HAc/HBu and HPr/HVa occurred respectively at the first and second biogas peaks. Biogas yield increased by 20 % after zeolite addition, about 356 mL/g VS_added with accelerated soluble chemical oxygen demand degradation and VFA (especially HPr and HBu) consumption in addition to a shortened lag phase between the two biogas peaks. Compared with Ca²⁺ and Mg²⁺ (100–300 mg/L) released from zeolite, simultaneous K⁺ and NH₄ ⁺ (580–600 mg/L) adsorptions onto zeolite particles contributed more to the enhanced biogasification, resulting in alleviated inhibition effects of ammonium on acidogenesis and methanogenesis, respectively. All the identified anaerobes could be grouped into Bacteroidetes and Firmicutes, and zeolite addition had no significant influence on the microbial biodiversity in this study.     

Anaerobic degradation of veratrylglycerol-beta-guaiacyl ether and guaiacoxyacetic acid by mixed rumen bacteria

Veratrylglycerol-beta-guaiacyl ether (0.2 g/liter), a lignin model compound, was found to be degraded by mixed rumen bacteria in a yeast extract medium under strictly anaerobic conditions to the extent of 19% within 24 h. Guaiacoxyacetic acid, 2-(o-methoxyphenoxy)ethanol, vanillic acid, and vanillin were detected as degradation products of veratrylglycerol-beta-guaiacyl ether by thin-layer chromatography, gas chromatography, and gas chromatography-mass spectrometry. Guaiacoxyacetic acid (0.25 g/liter), when added into the medium as a substrate, was entirely degraded within 36 h, resulting in the formation of phenoxyacetic acid, guaiacol, and phenol. These results suggest that the beta-arylether bond, an important intermonomer linkage in lignin, can be cleaved completely by these rumen anaerobes.     

Aerotolerance of strictly anaerobic microorganisms and factors of defense against oxidative stress: a review

Effects of aerobic conditions on strictly anaerobic microorganisms belonging to diverse taxa (clostridia, acetogenic bacteria, lactic acid bacteria, bacteroids, sulfate-reducing bacteria, and methanogenic archaea) and differing considerably in their oxygen resistance have been reviewed, with emphasis on the role of aerotolerance in the ecology of anaerobes. Consideration is given to components of nutritive media for anaerobe culturing, which decrease the toxic effects of oxygen and there by contribute significantly to maintenance and storage of industrial cultures of strictly anaerobic microorganisms. Physiological and biochemical factors are described, accounting for the relative resistance of many strict anaerobes to oxygen and products of incomplete reduction thereof. Specific attention is given to regulation of enzymes of antioxidative defense, operating in the cells of strict anaerobes under the conditions of oxidative stress caused by oxygen, superoxide anion, or hydrogen peroxide.     

Aerotolerance of strictly anaerobic microorganisms and factors of defense against oxidative stress: A review

Effects of aerobic conditions on strictly anaerobic microorganisms belonging to diverse taxa (clostridia, acetogenic bacteria, lactic acid bacteria, bacteroids, sulfate-reducing bacteria, and methanogenic archaea) and differing considerably in their oxygen resistance have been reviewed, with emphasis on the role of aerotolerance in the ecology of anaerobes. Consideration is given to components of nutritive media for anaerobe culturing, which decrease the toxic effects of oxygen and there by contribute significantly to maintenance and storage of industrial cultures of strictly anaerobic microorganisms. Physiological and biochemical factors are described, accounting for the relative resistance of many strict anaerobes to oxygen and products of incomplete reduction thereof. Specific attention is given to regulation of enzymes of antioxidative defense, operating in the cells of strict anaerobes under the conditions of oxidative stress caused by oxygen, superoxide anion, or hydrogen peroxide.     

Anaerobic degradation of veratrylglycerol-beta-guaiacyl ether and guaiacoxyacetic acid by mixed rumen bacteria.

Veratrylglycerol-beta-guaiacyl ether (0.2 g/liter), a lignin model compound, was found to be degraded by mixed rumen bacteria in a yeast extract medium under strictly anaerobic conditions to the extent of 19% within 24 h. Guaiacoxyacetic acid, 2-(o-methoxyphenoxy)ethanol, vanillic acid, and vanillin were detected as degradation products of veratrylglycerol-beta-guaiacyl ether by thin-layer chromatography, gas chromatography, and gas chromatography-mass spectrometry. Guaiacoxyacetic acid (0.25 g/liter), when added into the medium as a substrate, was entirely degraded within 36 h, resulting in the formation of phenoxyacetic acid, guaiacol, and phenol. These results suggest that the beta-arylether bond, an important intermonomer linkage in lignin, can be cleaved completely by these rumen anaerobes.     

Response surface methodology analysis of anaerobic syntrophic degradation of volatile fatty acids in an upflow anaerobic sludge bed reactor inoculated with enriched cultures

Anaerobic oxidation of volatile fatty acids (VFAs) as the key intermediates is restricted thermodynamically. Presently, enriched acetogenic and methanogenic cultures were used for syntrophic anaerobic digestion of VFAs in an upflow anaerobic sludge bed reactor fed with acetic, propionic, and butyric acids at maximum concentrations of 5.0, 3.0, and 4.0 g/L, respectively. Interactive effects of propionate, butyrate and acetate were analyzed. Hydraulic retention time (HRT) and acetate oxidizing syntrophs and methanogen (hydrogenotrophs) to syntrophic bacteria (propionate- and butyrate-oxidizing bacteria) population ratio (M/A) were investigated as key microbiological and operating variables of VFA anaerobic degradations. M/A did not affect the size distribution and had little effect on extracellular polymer contents of the granules. Granular sludge with close spatial microbial proximity enhanced syntrophic degradation of VFAs compared to other cultures, such as suspended cultures. Optimum conditions were found to be propionate = 1.93 g/L, butyrate = 2.15 g/L, acetate = 2.50 g/L, HRT = 22 h, and M/A = 2.5 corresponding to maximum VFA removal and biogas production rate. Results of verification experiments and predicted values from fitted correlations were in close agreement at the 95% confidence interval. Granules seemed to be smaller particles and less stable in construction with an irregular fractured surface compared to the original granules.     

Anaerobic microbiology in 198 cases of pleural empyema: a Bulgarian study

The aim of the study was to evaluate the incidence of anaerobic bacteria in 198 patients with pleural empyema and the susceptibility of isolates to eight antibacterial agents. Isolates were identified by the Crystal anaerobes identification system, API System rapid ID 32 A and/or routine methods. Susceptibility was tested by Sceptor MIC system for anaerobic bacteria and limited agar dilution method. Anaerobic bacteria were found in 74.2% of the patients and included 247 strains within 21 genera. The predominant anaerobes were Gram-positive anaerobic cocci (52 isolates), Fusobacterium (51), microaerophilic streptococci (24), Prevotella (19) and Bacteroides species (11). Common species/groups were Fusobacterium nucleatum (in 27.2% of specimens yielding anaerobes), Micromonas micros (8.2%), Finegoldia magna (7.5%), Bacteroides fragilis group (6.8%), Peptostreptococcus anaerobius (6.1%) and F. necrophorum (5.4%). No resistance to chloramphenicol and ampicillin/sulbactam was detected. The susceptibility rates of Gram-negative anaerobic isolates to penicillin, cefoxitin, clindamycin, clarithromycin, metronidazole and tetracycline were 63.8%, 90.2%, 87.8%, 58.6%, 98.8% and 71%, and those of Gram-positive anaerobes were 79.2%, 100%, 84.3%, 68.4%, 41.9% and 75%, respectively. The wide diversity of isolated anaerobic genera and species and the susceptibility patterns of the isolates emphasize the role of the anaerobic microbiology in cases of pleural empyema.     

Volatile fatty acid production from lignocellulosic biomass by lime pretreatment and its applications to industrial biotechnology

Lignocellulosic biomass was pretreated with lime, and used for the production of VFA (volatile fatty acid) through batch anaerobic digestion. About 0.34 g VFA yield was obtained using 10 g/L reed, after 3 days of fermentation with lime treatment; however, a higher VFA yield (more than 0.5 g/g biomass) was achieved with a modified lime treatment. Overall, our study showed that that the modified lime treatment is better suited for VFA production. VFAs can be widely used in platforms for fuels and chemicals from biomass.     

Stability of a percolating anaerobic downflow fixed film reactor under overloading conditions

Summary An anaerobic downflow fixed film (DSFF) reactor was used to treat cheese whey. The reactor was initially operated flooded (DSFF mode) and then the liquid level reduced by 67% (ATF mode); thus, the packing support was completely submerged and one third submerged respectively. The partly submerged anaerobic culture had higher gas production, and was more resistant to general and localized high VFA levels; methanogenic bacteria were acclimated to VFA levels as high as 3000 mg/l of propionate and 1600 mg/l of acetate (pH near neutrality) whilst the reactor remained stable.     

啤酒废水的厌氧发酵产氢

对经热处理后的厌氧污泥利用啤酒废水厌氧产氢的影响因素（温度、初始pH值和有机物浓度）进行了研究。结果表明,温度与初始pH值对厌氧产氢过程均有显著影响。最佳温度为35℃,此时,比产氢速率、氢气产率、VFA含量与总糖降解率均达到最大,分别为10.16mL/g-VSS.h、0.1673mL/mg.COD、4640.0mg/L和95.20%。最适初始pH值为6.0～7.0,在此范围内氢气产率、VFA含量、总糖降解率均可获得最大值。一个半经验模型适用于描述初始pH值与氢气产率之间的相互关系。在35℃、初始pH值6.5的条件下,有机物浓度COD1000mg/L～2000mg/L时,总糖降解率与VFA产率均获得最大值。底物抑制模型分析结果显示COD1587mg/L时,氢气产率（0.1935mL/mg.COD）达到最大。     

Incorporation of [methyl-3H]thymidine by obligate and facultative anaerobic bacteria when grown under defined culture conditions

Abstract Incorporation of [ methyl -³H]thymidine into bacterial DNA was determined for a range of axenic anaerobic bacterial cultures: fermentative heterotrophs, sulphate-reducing bacteria, purple sulphur bacteria, acetogens and methanogens. Anaerobically growing Bacillus sp. and the obligate aerobe Thiobacillus ferrooxidans were also investigated. Actively growing cultures of sulphate-reducing bacteria belonging to the genera Desulfovibrio, Desulfotomaculum, Desulfobacter, Desulfobotulus and Desulfobulbus , purple sulphur bacteria ( Chromatium vinosum OP2 and Thiocapsa roseopersicina OP1), methanogens ( Methanococcus GS16 and Methanosarcina barkeri ) and an acetogen ( Acetobacterium woodii ) did not incorporate [ methyl -³H]thymidine into DNA. The only obligate anaerobes in which thymidine incorporation into DNA could be unequivocally demonstrated were members of the genus Clostridium . Anaerobically growing Bacillus sp. also incorporated thymidine. These data demonstrate that pure culture representatives of major groups of anaerobic bacteria involved in the terminal oxidation of organic carbon and anoxygenic phototrophs within sediments are unable to incorporate [ methyl -³H]thymidine into DNA, although some obligate and facultative anaerobes can. Variability in thymidine incorporation amongst pure culture isolates indicates that unless existing techniques can be calibrated to take this into consideration then productivity estimates in both aerobic and anaerobic environments may be greatly underestimated using the [ methyl -³H]thymidine technique.     

Effect of temperature and hydraulic retention time on volatile fatty acid production based on bacterial community structure in anaerobic acidogenesis using swine wastewater

To investigate the effect of hydraulic retention time (HRT) and temperature (T) on bacterial community structure and volatile fatty acids (VFAs) production of an acidogenic process, and VFA production and changes in the bacterial community in three identical automated anaerobic continuously-stirred tank reactors were analyzed using response surface analysis (RSA) and nonmetric multidimensional scaling (NMDS). For RSA, 11 trials were conducted to find the combination of T and HRT under which VFA production was greatest; VFA production was affected more by HRT than by T. To identify the bacterial community structure in each trial, DNA from each experimental point of the RSA was analyzed using denaturating gradient gel electrophoresis (DGGE), and eight bacteria species were detected. NMDS was conducted on band intensities obtained using DGGE, and bacterial community structure was affected more by T than by HRT. Taken together, these results suggest that VFA production during acidogenesis was more dependent on the physicochemical properties of acidogens, such as their specific growth rate or contact time with of substrates, than on changes in the microbial community.     

Utilization of volatile fatty acids from the anaerobic digestion liquor of sewage sludge for 5-aminolevulinic acid production by photosynthetic bacteria

Using volatile fatty acids (VFA) from the anaerobic digestion liquor of sewage sludge, up to 9.2 mm 5-aminolevulinic acid (ALA) could be produced by Rhodobacter sphaeroides under anaerobic-light (5 kLux) conditions with repeated addition of levulinic acid (LA) and glycine and using a large inoculum (approx. 2 g/l of cells, initially from glutamate/malate medium). As the VFA medium also contained organic nitrogen sources such as glutamic acid, the cells were later grown up to about 2 g/l in the VFA medium instead of the glutamate/malate medium. ALA production was then again promoted by adding LA and glycine. Using this improved method, up to 9.3 mm ALA was produced by feeding propionate and acetate together with LA and glycine, indicating that VFA medium formed from sewage sludge could be useful for ALA production.     

Growth of a facultative anaerobe under oxygen-limiting conditions in pure culture and in co-culture with a sulfate-reducing bacterium

Abstract The occurrence and properties were studied of glucose-metabolizing bacteria present in the anaerobic sediment 5–10 cm below the surface of an estuarine tidal mud-flat. Of all these bacteria (10⁴– 10⁵ per g wet sediment) 80–90% were facultatively anaerobic species. Chemostat enrichments on glucose under aerobic, oxygen-limited and alternately aerobic-anaerobic conditions also yielded cultures dominated by facultative anaerobes. One of the dominant species, tentatively identified as a Vibrio sp., was studied in more detail under oxygen-limiting conditions. Fermentative and respiratory metabolisms were found to operate simultaneously, and the ratio between the two was regulated by the extent of oxygen limitation. A small fraction of the acetate formed under such growth conditions was shown to be subsequently respired. A co-culture was established of the Vibrio sp. and a sulfate-reducing bacterium ( Desulfovibrio HL21 ) in an aerated chemostat. The importance of these observations is discussed in relation to the role of facultative anaerobes in anaerobic habitats.     

Anaerobes in the Nasopharynx During Acute Otitis Media Episodes in Infancy

The present study, as a part of our longitudinal study on the development of the nasopharyngeal microflora, investigates the nasopharyngeal carriage of anaerobes during 75 episodes of acute otitis media (AOM) experienced by 34 infants during their first 2 years of life. Special emphasis was put on adequate methods of detecting anaerobic bacteria from carefully collected nasopharyngeal aspirates. During the follow-up period, two-thirds of the infants carried anaerobes in the nasopharynx at least once. Anaerobic species were recovered from 47% of the nasopharyngeal aspirates. Environmental/ecological changes in the nasopharynx during infection favour the growth of anaerobes. Their significance in the pathogenesis of AOM, if any, warrants further study.     

细菌作为抗肿瘤剂的研究进展

厌氧和兼性厌氧细菌能够专一性地优先累积在肿瘤的低氧量和坏死的区域,这使其具有很好的靶向性。它们能够分泌毒素或水解酶以及引起针对肿瘤的免疫反应,具有溶瘤作用。基因工程技术能够改造这些细菌使其具有抗肿瘤的特性,进一步增加治疗效果。最新的研究发现,这些细菌的某些代谢产物也具有增强肿瘤治疗效果的作用,为肿瘤的治疗提供了新的思路。本文对细菌作为抗肿瘤剂的相关研究进行综述。     

Using enriched cultures for elevation of anaerobic syntrophic interactions between acetogens and methanogens in a high-load continuous digester

Volatile fatty acids (VFAs) are key intermediates in anaerobic digestion. Enriched acetogenic and methanogenic cultures were used for syntrophic anaerobic digestion of VFAs in a high-load continuous reactor fed with acetic (HAc), propionic (HPr) and butyric (HBu) acids at maximum concentrations of 5, 3 and 4 g/L, respectively. Interactive effects of HPr, HBu and HAc were analyzed. Furthermore, hydraulic retention time (HRT) and methanogen to acetogen population ratio (M/A) were investigated as key microbiological and operating variables of VFA anaerobic degradations. Optimum conditions were found to be HPr = 1125.0 mg/L, HBu = 1833.4 mg/L, HAc = 1727.4 mg/L, HRT = 21 h and M/A = 2.5 (corresponding to maximum VFA removal and biogas production rate (BPR)). Results of verification experiments and predicted values from fitted correlations were in close agreement at 95% confidence interval. HRT and M/A had positive effects on VFA removal and BPR. M/A was the most important factor that affected BPR. All VFAs inhibited VFA removals.