Degradation of Alpha-, Beta-, and Gamma-Hexachlorocyclohexane by a Soil Bacterium under Aerobic Conditions

A Pseudomonas sp., isolated from sugarcane rhizosphere soil, readily metabolized not only alpha and gamma isomers of hexachlorocyclohexane, but also the thermodynamically more stable beta isomer, under aerobic conditions. Bacterial degradation of the three isomers led to the accumulation of a transitory metabolite and eventual release of covalently linked chlorine as chloride in stoichiometric amounts.     

Ethylene Dibromide Mineralization in Soils under Aerobic Conditions

1,2-Dibromoethane (EDB), which is a groundwater contaminant in areas where it was once used as a soil fumigant, was shown to be degraded aerobically by microorganisms in two types of surface soils from an EDB-contaminated groundwater discharge area. At initial concentrations of 6 to 8 μg/liter, EDB was degraded in a few days to near or below the detection limit of 0.02 μg/liter. At 15 to 18 mg/liter, degradation was slower. Bromide ion release at the higher concentrations was 1.4 ± 0.3 and 2.1 ± 0.2 molar equivalents for the two soils. Experiments with [¹⁴C]EDB showed that EDB was converted to approximately equal amounts of CO₂ and apparent cellular carbon; only small amounts of added ¹⁴C were not attributable to these products or unreacted EDB. These results are encouraging, because they indicate that groundwater bacteria may hasten the removal of EDB from contaminated aerobic groundwater supplies. This report also provides evidence for soil-mediated chemical transformations of EDB.     

Impact of an Aerobic Thermophilic Sequencing Batch Reactor on Antibiotic-Resistant Anaerobic Bacteria in Swine Waste

The introduction of antibiotics to animal feed has contributed to the selection of antibiotic-resistant bacteria in concentrated animal feeding operations. The aim of this work was to characterize the impact of an aerobic thermophilic biotreatment on anaerobic antibiotic-resistant bacteria in swine waste. Despite 162- to 6,166-fold reduction in antibiotic-resistant populations enumerated in the swine waste at 25°C and 37°C, resistant populations remained significant (10⁴ to 10⁵ most probable number per milliliter) in the treated swine waste. Five resistance genes were detected before [tet(LMOS) erm(B)], and six resistance genes were detected after [tet(LMOSY) erm(B)] biotreatment. However, the biotreatment decreased the frequency of detection of resistance genes by 57%. Analysis by denaturing gradient gel electrophoresis of polymerase chain reaction-amplified 16 S ribosomal DNA (rDNA) fragments showed that the biotreatment reduced the bacterial diversity of resistant populations enumerated at 37°C. Cloning and sequencing of the 16 S rDNA of these populations revealed that most clones in the treated swine waste were closely similar to some of the clones retrieved from the untreated swine waste. This study revealed that the aerobic thermophilic biotreatment developed in our laboratory does not prevent the introduction of facultatively anaerobic antibiotic-resistant bacteria and their resistance genes into agricultural ecosystems. Horizontal transfer of ecologically advantageous genes within microbial communities are likely to prevent thermophilic biotreatments from completely eliminating antibiotic-resistant bacteria and their resistance genes in animal wastes.     

Degradation of Microbicides under Different Environmental Conditions

To determine the biodegradability of some widely used microbicides, these products were dissolved in a mineral solution and synthetic sewage and their breakdown was followed under aerobic and anaerobic conditions. The results indicated that most biocides will not raise pollution problems provided they are discharged into domestic sewage and subsequently treated by aerobic water purification systems. However, all but a few of the biocides tested failed to be degraded in tests simulating conditions prevailing in oligotrophic aquatic systems or in anaerobic ecosystems.     

Acetic Acid Increases Stability of Silage under Aerobic Conditions

The effects of various compounds on the aerobic stability of silages were evaluated. It has been observed that inoculation of whole-crop maize with homofermentative lactic acid bacteria leads to silages which have low stability against aerobic deterioration, while inoculation with heterofermentative lactic acid bacteria, such as Lactobacillus brevis or Lactobacillus buchneri, increases stability. Acetic acid has been proven to be the sole substance responsible for the increased aerobic stability, and this acid acts as an inhibitor of spoilage organisms. Therefore, stability increases exponentially with acetic acid concentration. Only butyric acid has a similar effect. Other compounds, like lactic acid, 1,2-propanediol, and 1-propanol, have been shown to have no effect, while fructose and mannitol reduce stability.     

Degradation of Polycyclic Aromatic Hydrocarbons at Low Temperature under Aerobic and Nitrate-Reducing Conditions in Enrichment Cultures from Northern Soils

The potential for biodegradation of polycyclic aromatic hydrocarbons (PAHs) at low temperature and under anaerobic conditions is not well understood, but such biodegradation would be very useful for remediation of polluted sites. Biodegradation of a mixture of 11 different PAHs with two to five aromatic rings, each at a concentration of 10 μg/ml, was studied in enrichment cultures inoculated with samples of four northern soils. Under aerobic conditions, low temperature severely limited PAH biodegradation. After 90 days, aerobic cultures at 20°C removed 52 to 88% of the PAHs. The most extensive PAH degradation under aerobic conditions at 7°C, 53% removal, occurred in a culture from creosote-contaminated soil. Low temperature did not substantially limit PAH biodegradation under nitrate-reducing conditions. Under nitrate-reducing conditions, naphthalene, 2-methylnaphthalene, fluorene, and phenanthrene were degraded. The most extensive PAH degradation under nitrate-reducing conditions at 7°C, 39% removal, occurred in a culture from fuel-contaminated Arctic soil. In separate transfer cultures from the above Arctic soil, incubated anaerobically at 7°C, removal of 2-methylnaphthalene and fluorene was stoichiometrically coupled to nitrate removal. Ribosomal intergenic spacer analysis suggested that enrichment resulted in a few predominant bacterial populations, including members of the genera Acidovorax, Bordetella, Pseudomonas, Sphingomonas, and Variovorax. Predominant populations from different soils often included phylotypes with nearly identical partial 16S rRNA gene sequences (i.e., same genus) but never included phylotypes with identical ribosomal intergenic spacers (i.e., different species or subspecies). The composition of the enriched communities appeared to be more affected by presence of oxygen, than by temperature or source of the inoculum.     

Fate of Chlortetracycline- and Tylosin-Resistant Bacteria in an Aerobic Thermophilic Sequencing Batch Reactor Treating Swine Waste

Antibiotics have been added to animal feed for decades. Consequently, food animals and their wastes constitute a reservoir of antibiotic-resistant bacteria. The objective of this work was to characterize the impact of an aerobic thermophilic biotreatment on aerobic, antibiotic-resistant bacteria in swine waste. The proportion of tylosin- and chlortetracycline-resistant bacteria grown at 25°C, 37°C, and 60°C decreased after treatment, but they were still abundant (10² to 10⁸ most probable number ml⁻¹) in the treated swine waste. The presence of 14 genes conferring resistance to tylosin and chlortetracycline was assessed by polymerase chain reaction in bacterial populations grown at 25°C, 37°C, and 60°C, with or without antibiotics. In 22 cases, genes were detected before but not after treatment. The overall gene diversity was wider before [tet(BLMOSY), erm(AB)] than after [tet(LMOS), erm(B)] treatment. Analysis by denaturing gradient gel electrophoresis of amplified 16S ribosomal DNA (rDNA) fragments generally showed a reduction of the bacterial diversity, except for total populations grown at 60°C and for tylosin-resistant populations grown at 37°C. The latter were further investigated by cloning and sequencing their 16S rDNA. Phylotypes found before treatment were all closely related to Enterococcus hirae, whereas six different phylotypes, related to Pseudomonas, Alcaligenes, and Pusillimonas, were found after treatment. This work demonstrated that the aerobic thermophilic biotreatment cannot be considered as a means for preventing the dissemination of aerobic antibiotic-resistant bacteria and their resistance genes to the environment. However, since pathogens do not survive the biotreatment, the effluent does not represent an immediate threat to animal or human health.     

Aerobic Biodegradation of Liquid Motor Fuels under Extreme Acidic Conditions

Microbiology - Biodegradation of liquid petroleum motor fuels and fuel mixtures containing biodiesel fuel (methyl ethers of rapeseed fatty acids) by aerobic acidophilic actinobacteria Mycobacterium...     

Amylolytic Activities in Cereal Seeds under Aerobic and Anaerobic Conditions

An adequate carbohydrate supply contributes to the survival of seeds under conditions of limited oxygen availability. The amount of soluble, readily fermentable carbohydrates in dry cereal seeds is usually very limited, with starch representing the main storage compound. Starch breakdown during the germination of cereal seeds is the result of the action of hydrolytic enzymes and only through the concerted action of [alpha]-amylase (EC 3.2.1.1), [beta]-amylase (EC 3.2.1.2), debranching enzyme (EC 3.2.1.41), and [alpha]-glucosidase (EC 3.2.1.20) can starch be hydrolyzed completely. We present here data concerning the complete set of starch-degrading enzymes in three cereals, rice (Oryza sativa L.), which is tolerant to anaerobiosis, and wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.), which are unable to germinate under anoxia. Among the cereal seeds tested under anoxia, only rice is able to degrade nonboiled, soluble starch, reflecting the ability to degrade the starch granules in vivo. This is explained by the presence of the complete set of enzymes needed to degrade starch completely either as the result of de novo synthesis ([alpha]-amylase, [beta]-amylase) or activation of preexisting, inactive forms of the enzyme (debranching enzyme, [alpha]-glucosidase). These enzymes are either absent or inactive in wheat and barley seeds kept under anaerobic conditions.     

Biofilm Formation by Campylobacter jejuni Is Increased under Aerobic Conditions

The microaerophilic human pathogen Campylobacter jejuni is the leading cause of food-borne bacterial gastroenteritis in the developed world. During transmission through the food chain and the environment, the organism must survive stressful environmental conditions, particularly high oxygen levels. Biofilm formation has been suggested to play a role in the environmental survival of this organism. In this work we show that C. jejuni NCTC 11168 biofilms developed more rapidly under environmental and food-chain-relevant aerobic conditions (20% O₂) than under microaerobic conditions (5% O₂, 10% CO₂), although final levels of biofilms were comparable after 3 days. Staining of biofilms with Congo red gave results similar to those obtained with the commonly used crystal violet staining. The level of biofilm formation by nonmotile aflagellate strains was lower than that observed for the motile flagellated strain but nonetheless increased under aerobic conditions, suggesting the presence of flagellum-dependent and flagellum-independent mechanisms of biofilm formation in C. jejuni. Moreover, preformed biofilms shed high numbers of viable C. jejuni cells into the culture supernatant independently of the oxygen concentration, suggesting a continuous passive release of cells into the medium rather than a condition-specific active mechanism of dispersal. We conclude that under aerobic or stressful conditions, C. jejuni adapts to a biofilm lifestyle, allowing survival under detrimental conditions, and that such a biofilm can function as a reservoir of viable planktonic cells. The increased level of biofilm formation under aerobic conditions is likely to be an adaptation contributing to the zoonotic lifestyle of C. jejuni.Infection with Campylobacter jejuni is the leading cause of food-borne bacterial gastroenteritis in the developed world and is often associated with the consumption of undercooked poultry products (19). The United Kingdom Health Protection Agency reported more than 45,000 laboratory-confirmed cases for England and Wales in 2006 alone, although this is thought to be a 5- to 10-fold underestimation of the total number of community incidents (20, 43). The symptoms associated with C. jejuni infection usually last between 2 and 5 days and include diarrhea, vomiting, and stomach pains. Sequelae of C. jejuni infection include more-serious autoimmune diseases, such as Guillain-Barré syndrome, Miller-Fisher syndrome (18), and reactive arthritis (15).Poultry represents a major natural reservoir for C. jejuni, since the organism is usually considered to be a commensal and can reach densities as high as 1 × 10⁸ CFU g of cecal contents⁻¹ (35). As a result, large numbers of bacteria are shed via feces into the environment, and consequently, C. jejuni can spread rapidly through a flock of birds in a broiler house (1). While well adapted to life in the avian host, C. jejuni must survive during transit between hosts and on food products under stressful storage conditions, including high and low temperatures and atmospheric oxygen levels. The organism must therefore have mechanisms to protect itself from unfavorable conditions.Biofilm formation is a well-characterized bacterial mode of growth and survival, where the surface-attached and matrix-encased bacteria are protected from stressful environmental conditions, such as UV radiation, predation, and desiccation (7, 8, 28). Bacteria in biofilms are also known to be >1,000-fold more resistant to disinfectants and antimicrobials than their planktonic counterparts (11). Several reports have now shown that Campylobacter species are capable of forming a monospecies biofilm (21, 22) and can colonize a preexisting biofilm (14). Biofilm formation can be demonstrated under laboratory conditions, and environmental biofilms, from poultry-rearing facilities, have been shown to contain Campylobacter (5, 32, 44). Campylobacter biofilms allow the organism to survive up to twice as long under atmospheric conditions (2, 21) and in water systems (27).Molecular understanding of biofilm formation by Campylobacter is still in its infancy, although there is evidence for the role of flagella and gene regulation in biofilm formation. Indeed, a flaAB mutant shows reduced biofilm formation (34); mutants defective in flagellar modification (cj1337) and assembly (fliS) are defective in adhering to glass surfaces (21); and a proteomic study of biofilm-grown cells shows increased levels of motility-associated proteins, including FlaA, FlaB, FliD, FlgG, and FlgG2 (22). Flagella are also implicated in adhesion and in biofilm formation and development in other bacterial species, including Aeromonas, Vibrio, Yersinia, and Pseudomonas species (3, 23, 24, 31, 42).Previous studies of Campylobacter biofilms have focused mostly on biofilm formation under standard microaerobic laboratory conditions. In this work we have examined the formation of biofilms by motile and nonmotile C. jejuni strains under atmospheric conditions that are relevant to the survival of this organism in a commercial context of environmental and food-based transmission.     

Siderophore Production by Pseudomonas stutzeri under Aerobic and Anaerobic Conditions

The siderophore production of the facultative anaerobe Pseudomonas stutzeri, strain CCUG 36651, grown under both aerobic and anaerobic conditions, was investigated by liquid chromatography and mass spectrometry. The bacterial strain has been isolated at a 626-m depth at the Äspö Hard Rock Laboratory, where experiments concerning the geological disposal of nuclear waste are performed. In bacterial culture extracts, the iron in the siderophore complexes was replaced by gallium to facilitate siderophore identification by mass spectrometry. P. stutzeri was shown to produce ferrioxamine E (nocardamine) as the main siderophore together with ferrioxamine G and two cyclic ferrioxamines having molecular masses 14 and 28 atomic mass units lower than that of ferrioxamine E, suggested to be ferrioxamine D₂ and ferrioxamine X₁, respectively. In contrast, no siderophores were observed from anaerobically grown P. stutzeri. None of the siderophores produced by aerobically grown P. stutzeri were found in anaerobic natural water samples from the Äspö Hard Rock Laboratory.     

Biodegradation of Benzene by Halophilic and Halotolerant Bacteria under Aerobic Conditions

A highly enriched halophilic culture was established with benzene as the sole carbon source by using a brine soil obtained from an oil production facility in Oklahoma. The enrichment completely degraded benzene, toluene, ethylbenzene, and xylenes within 1 to 2 weeks. Also, [¹⁴C]benzene was converted to ¹⁴CO₂, suggesting the culture's ability to mineralize benzene. Community structure analysis revealed that Marinobacter spp. were the dominant members of the enrichment.     

Interactions of Nitric Oxide with Lipid Peroxidation Products under Aerobic Conditions: Inhibitory Effects on the Formation of Malondialdehyde and Related Thiobarbituric Acid-Reactive Substances

Under aerobic conditions, exposure of peroxidized lipids to nitric oxide (NO) was found to result in a rapid decrease in the levels of thiobarbituric acid-reactive substances (TBARS). Addition of 10–100 μM NO to rat brain homogenates preincubated for 2 h at 37°C caused up to a 20% decrease in the levels of TBARS compared to controls. A similar inhibitory effect was observed on TBARS produced by Fe²⁺-induced decomposition of 15-hydroperoxyeicosatetraenoic acid (15-HPETE), due apparently to NO-induced decomposition of the hydroperoxide (ferrous oxidation/xylenol orange assay). Prostaglandin G₂ (PGG₂, 35 μM), as a model bicyclic endoperoxide, and malondialdehyde (MDA, 20 μM), the main component of TBARS, proved also susceptible to degradation by NO or NO donors (diethylamine NONOate, DEA/NO) at concentrations of 100 μM or higher in 0.05 M phosphate buffer, pH 7.4, and at 37°C, as indicated by the reduced response to the TBA assay. No significant effect on TBARS determination was caused by nitrite ions. These and other data indicate that NO can inhibit TBARS formation by decomposing primary lipid peroxidation products, chiefly 15-HPETE and related hydroperoxides, and, to a lesser extent, later stage TBARS precursors, including bicyclic endoperoxides and MDA, via nitrosation and other oxidative routes, without however affecting chromogenic reactions during the assay.     

Procurement and Maintenance of Germ-Free Swine for Microbiological Investigations

Germ-free swine were routinely procured by both hysterectomy and hysterotomy (Caesarian section). By means of light-weight portable equipment, piglets could be obtained and transported to the laboratory (without contamination) over distances in excess of 100 miles. The isolators employed in rearing were constructed of stainless steel and flexible plastic film. At weekly intervals, fecal swabs and waste from the floor of the isolator were cultured on blood-agar and in thioglycolate broth, as well as being examined microscopically for the presence of bacteria, yeast, and fungi. The presence of pleuropneumonia-like organisms (PPLO) and viruses in such material was not demonstrable, either by the use of enriched PPLO media or primary porcine-kidney cell cultures. Tissues, body fluids, and cecal contents of piglets sacrificed specifically for microbiological examination were also negative for PPLO, viruses, bacteria, yeast, and fungi. Prenatal infestations by ascarids were not observed. Nutritional problems related to rearing of germ-free piglets, such as hypoglycemia, were not encountered, and the use of an autoclaved commercial sow's milk replacer proved quite satisfactory. The temperature to which piglets were subjected during the first few days of life, however, was very important. The isolator design and application of gnotobiotic techniques to the procurement and rearing of a large germ-free animal such as the pig proved feasible and less difficult than anticipated.     

Aerobic Microbial Degradation of Glucoisosaccharinic Acid

α-Glucoisosaccharinic acid (GISA), a major by-product of kraft paper manufacture, was synthesized from lactose and used as the carbon source for microbial media. Ten strains of aerobic bacteria capable of growth on GISA were isolated from kraft pulp mill environments. The highest growth yields were obtained with Ancylobacter spp. at pH 7.2 to 9.5. GISA was completely degraded by cultures of an Ancylobacter isolate. Ancylobacter cell suspensions consumed oxygen and produced carbon dioxide in response to GISA addition. A total of 22 laboratory strains of bacteria were tested, and none was capable of growth on GISA. GISA-degrading isolates were not found in forest soils.     

Attachment Behavior of Shewanella putrefaciens onto Magnetite under Aerobic and Anaerobic Conditions

In this study, batch experiments were used to characterize attachment behavior of Shewanella putrefaciens strain 200R to ferrihydrite and magnetite. Attachment was quantified in batch experiments with a 0.01 M NaNO ₃ solution as a function of pH (ranging from 3 to 10), sorbed anion (PO₄ ^{3 ?} ), and growth conditions (aerobic vs. anaerobic). Electrophoretic mobility data was collected for S. putrefaciens cells and magnetite grains and used as a means to interpret the role of electrostatic interaction in attachment studies. Little difference in attachment behavior was observed as a function of growth conditions or surface treatments. The exception was at pH ranging from 2 to 4, under anaerobic conditions, where increased attachment was measured on magnetite surfaces with sorbed PO₄ ^{3 ?} . This increased attachment was attributed to development of Fe-PO₄ surface complexes or secondary mineral phases, resulting in altered surface interactions between cell and mineral surfaces. Attachment was irreversible and increased with time under anaerobic conditions even under elevated pH conditions unfavourable to electrostatic interactions between cells and mineral surfaces. These results suggest that electrophoretic mobility data in this system is not a good predictor of attachment behavior, while surface charge development via protonation and deprotonation of surface functional groups is consistent with experimental attachment data. In this study, S. putrefaciens appears to utilize polymers or pili to remain attached to Fe-oxides and this process may facilitate Fe reduction on these surfaces. Results from this study underscore the need for quantitative bulk measurements of microbial attachment to accurately predict partitioning of dissimilatory iron reducing bacteria between solution and solid phases.     

Production of a Gaseous Saturated-hydrocarbon Mixture by Rhizopus japonicus under Aerobic Conditions

The microbial production, by the genus Rhizopus, of a gaseous saturated-hydrocarbon mixture was studied under aerobic conditions. Rhizopus strains, comprising 13 strains of 9 species, were tested as to their ability to produce a gaseous hydrocarbon mixture. Except for one strain, all the strains tested produced more than two kinds of gaseous hydrocarbons simultaneously when grown in nutrient broth containing glucose. Rhizopus japonicus IFO 4758 was selected as being typical of these producers of mixed gaseous hydrocarbons. When this organism was cultivated in a synthetic medium supplemented with l-cysteine under aerobic conditions, the maximum production of the total gaseous hydrocarbon mixture reached ca. 10 nl/ml culture broth/hr. The gaseous hydrocarbon mixture produced was composed mainly of paraffin hydrocarbons, i.e., ca. 74% pentane, ca. 16% propane and a trace amount of methane. The ratios of saturated to unsaturated, and even to odd number hydrocarbons produced by this fungus were 95 : 5 and 90: 10, respectively. The biosynthetic pathways for the production of these gases are discussed in comparison with the biosynthetic pathways for ethylene and isobutene in microorganisms.     

城市生活垃圾的微生物处理技术

城市生活垃圾量的日益增多严重威胁着城市的环境及人们的健康,垃圾处理成为现代社会面临的重要问题之一,因此城市生活垃圾处理技术对于城市的可持续发展起着非常重要的作用。微生物在自然界中种类繁多,在城市生活垃圾生物处理应用中占有重要地位。本文对微生物在城市生活垃圾的卫生填埋技术、生物反应器填埋技术、好氧堆肥技术、发酵降解技术及生物干燥器技术中的应用进行了综述,并对城市生活垃圾的综合处理提出建议,以期为城市生活垃圾的微生物处理技术的应用提供参考。