Acetate threshold values and acetate activating enzymes in methanogenic bacteria

Abstract The minimum threshold concentrations of acetate utilization and the enzymes responsible for acetate activation of several methanogenic bacteria were investigated and compared with literature data. The minimum acetate concentrations reached by hydrogenotrophic methane bacteria, which require acetate as carbon source, were between 0.4 and 0.6 mM. The acetoclastic Methanosarcina achieves acetate concentrations between 0.2 and 1.2 mM and Methanothrix between 7 and 70 μM. For the activation of acetate most of the hydrogenotrophic methane bacteria investigated use an acetyl-CoA synthetase with a relatively low K _m (40–90 μM) for acetate. although the affinity for acetate was high, the hydrogenotrophic methane bacteria were not able to remove acetate to lower concentrations than the acetoclastic methane bacteria, neither in pure cultures nor in anaerobic granular sludge samples. Based on these observations, it is not likely that hydrogenotrophic methanogens compete strongly for acetate with the acetoclastic methane bacteria.     

Heterotrophic bacterial production on solid fish waste: TAN and nitrate as nitrogen source under practical RAS conditions

The drumfilter effluent from a recirculation aquaculture system (RAS) can be used as substrate for heterotrophic bacteria production. This biomass can be re-used as aquatic feed. RAS effluents are rich in nitrate and low in total ammonia nitrogen (TAN). This might result in 20% lower bacteria yields, because nitrate conversion into bacteria is less energy efficient than TAN conversion. In this study the influence of TAN concentrations (1, 12, 98, 193, 257mgTAN/l) and stable nitrate-N concentrations (174+/-29mg/l) on bacteria yields and nitrogen conversions was investigated in a RAS under practical conditions. The effluent slurry was supplemented with 1.7gC/l sodium acetate, due to carbon deficiency, and was converted continuously in a suspended bacteria growth reactor (hydraulic retention time 6h). TAN utilization did not result in significantly different observed yields than nitrate (0.24-0.32gVSS/gC, p=0.763). However, TAN was preferred compared to nitrate and was converted to nearly 100%, independently of TAN concentrations. TAN and nitrate conversions rates were differing significantly for increasing TAN levels (p<0.000 and p=0.012), and were negatively correlated. It seems, therefore, equally possible to supply the nitrogenous substrate for bacteria conversion as nitrate and not as TAN. The bacteria reactor can, as a result, be integrated into an existing RAS as end of pipe treatment.     

Bactericidal activity of the serum against Staphylococcus aureus grown in the presence of subinhibitory doses of cefamandole and gentamicin

Growth of S. aureus in the presence of subinhibitory concentrations of cefamandole alone (at 1/5 the MIC) or with gentamicin (both at 1/5 the MIC) produced large, globular cells with reduced viability (50%) as compared to untreated bacteria. Gentamicin (1/5 the MIC) did not modify bacteria and also reduced viability of inoculum (57% vs. controls). Cefamandole treated bacteria were more susceptible to the rat serum bactericidal activity then control bacteria. The growth of S. aureus in the presence of cefamandole plus gentamicin did not modify further bacterial susceptibility to serum opsonins. The data suggest that even at subinhibitory concentrations cefamandole may exert an antibacterial effect by acting in cooperation with serum factors.     

Soil: the environmental source of Escherichia coli and Enterococci in Guam's streams

We have previously documented that faecal indicator bacteria (Escherichia coli, faecal coliform, enterococci) recommended by the U.S. Environmental Protection Agency (USEPA) to establish recreational water quality standards are naturally found in high concentrations in the surface and subsurface of soils in Hawaii. Rain, the source of all streams in Hawaii, washes the soil sources of faecal bacteria into all the streams of Hawaii, at concentrations which consistently exceed the USEPA recreational water quality standards. The objective of this study was to test the hypothesis that faecal bacteria are able to establish themselves in the soil environments of tropical islands by conducting the same study in Guam, a tropical pacific island with warmer temperatures and higher humidity than Hawaii. The same methods and study design used in Hawaii was used in Guam. The results of the study conducted in Guam revealed that all streams contain consistently high concentrations of faecal coliform, E. coli, and enterocci which exceeded the old USEPA recreational water quality standard of 200 faecal coliform/100 ml as well as the new water quality standards of 126 E. coli/100 ml or 33 enterococci/100 ml. These same faecal indicator bacteria were recovered in high concentrations in surface and subsurface (18-36 cm depth) soil samples in Guam. Limited coastal water analysis showed that most coastal marine waters contain low concentrations of faecal bacteria but coastal waters impacted by stream run-off showed elevated levels of faecal bacteria. The results of this study support the hypothesis that environmental conditions in the tropical areas of the world can support the growth and establishment of populations of faecal bacteria in the soil. Thus, soil becomes an environmental, non-faecal source of faecal indicator bacteria. These results indicate that USEPA water quality standards may not be directly applicable to tropical island environments.     

Strong synergy between a eukaryotic antimicrobial peptide and bacteriocins from lactic acid bacteria

The antimicrobial effect obtained upon combining the prokaryotic antimicrobial peptides (AMPs; more commonly referred to as bacteriocins) pediocin PA-1, sakacin P, and curvacin A (all produced by lactic acid bacteria [LAB]) with the eukaryotic AMP pleurocidin (from fish) has been investigated. The three LAB AMPs alone were active against gram-positive Listeria ivanovii bacteria at nanomolar concentrations, whereas they were inactive against gram-negative Escherichia coli bacteria. Pleurocidin alone was active against both of these types of bacteria at micromolar concentrations. Little if any synergy between the LAB AMPs and pleurocidin against the gram-positive L. ivanovii strain was obtained. In contrast, the LAB AMPs and pleurocidin acted highly synergistically against the gram-negative E. coli strain. Nanomolar concentrations of LAB AMPs increased the growth inhibitory potency of pleurocidin by about fourfold. When micromolar concentrations of LAB AMPs were combined with 2 micro g of pleurocidin/ml, 100% growth inhibition was attained, whereas pleurocidin alone at a concentration of 2 micro g/ml gave no growth inhibition. Most noteworthy, when high concentrations (128 micro g/ml) of pleurocidin in the absence of LAB AMPs were used over a long period of incubation (1 week), some growth of E. coli was observed, whereas 16 micro g of pleurocidin/ml completely abolished growth in the presence of 64 to 128 ng of LAB AMPs/ml over the same period of time. The results clearly demonstrate that combining eukaryotic and prokaryotic AMPs can greatly increase the specific activity and broaden the target-cell range of these peptides.     

Complexing of copper in drinking water samples to enhance recovery of Aeromonas and other bacteria

The presence of copper in drinking water samples at concentrations as low as 10 micrograms/l resulted in a rapid die-off of aeromonads. Coliform bacteria, heterotrophic plate count bacteria and faecal streptococci were also sensitive to copper but to a lesser degree than aeromonads. The effect was particularly noticeable in soft water (less than 3 meq/l Ca + Mg) and at pH-values below 8.0. The toxic effect of copper concentrations up to 500 micrograms/l could be neutralized for a period of up to 24 h by the addition to samples of 50 mg/l of disodium-ethylene-diamino-tetraacetate (Na2EDTA) and keeping the samples on melting ice.     

Nitrite Removal Performance and Community Structure of Nitrite-Oxidizing and Heterotrophic Bacteria Suffered with Organic Matter

Altlhough ammonia oxidation and ammonia-oxidizing bacteria (AOB) have been extensively studied, nitrite oxidation and nitrite-oxidizing bacteria (NOB) are still not well understood. In this article, the effect of organic matter on NOB and heterotrophic bacteria was investigated with functional performance analysis and bacterial community shift analysis. The results showed that at low concentrations of initial sodium acetate [initial sodium acetate (ISA) = 0.5 or 1 g/L], the nitrite removal rate was higher than that obtained under autotrophic conditions and the bacteria had a single growth phase, whereas at high ISA concentrations (5 or 10 g/L), continuous aerobic nitrification and denitrification occurred in addition to higher nitrite removal rates, and the bacteria had double growth phases. The community structure of total bacteria strikingly varied with the different concentrations of ISA; the dominant populations shifted from autotrophic and oligotrophic bacteria (NOB, and some strains of Bacteroidetes, Alphaproteobacteria, Actinobacteria, and green nonsulfur bacteria) to heterotrophic and denitrifying bacteria (strains of Gammaproteobacteria, especially Pseudomonas stutzeri and P. nitroreducens). The reasons that nitrite removal rate increased with supplement of organic matters were discussed.     

Complexing of copper in drinking water samples to enhance recovery of Aeromonas and other bacteria

The presence of copper in drinking water samples at concentrations as low as 10 μg/l resulted in a rapid die-off of aeromonads. Coliform bacteria, heterotrophic plate count bacteria and faecal streptococci were also sensitive to copper but to a lesser degree than aeromonads. The effect was particularly noticeable in soft water (<3 meq/l Ca + Mg) and at pH-values below 8.0. The toxic effect of copper concentrations up to 500 μg/l could be neutralized for a period of up to 24 h by the addition to samples of 50 mg/l of disodium-ethylene-diamino-tetraacetate (Na₂EDTA) and keeping the samples on melting ice.     

六价铬对薏米人工湿地微生物群落数量的影响

通过桶栽构筑微型模拟垂直流薏米人工湿地(CAW),以1/2 Hoaglands营养液为营养源,在营养液中添加不同浓度的Cr6+(0,5,20,40,60mg/L,以K2Cr2O7配置),各浓度处理均以不种植薏米湿地(NPW)为对照,以研究铬(Cr6+)对薏米人工湿地基质真菌、细菌及放线菌群落结构数量的影响。结果表明:(1)真菌、细菌、放线菌的数量在薏米人工湿地(CAW)中明显多于无植物对照处理(NPW);(2)中低浓度(5、20mg/L)Cr6+对CAW真菌,对NPW细菌、放线菌数量有促进作用,薏米人工湿地微生物对低中浓度的Cr6+胁迫有一定的耐受能力。     

Effects of heavy metals and other trace elements on the fermentative activity of the rumen microflora and growth of functionally important rumen bacteria

The inhibitory effects of high concentrations of essential and non-essential trace elements were tested on the rumen microflora using the rate of fermentation in vitro as the assay. The elements (and the concentration causing 50% inhibition) in decreasing order of toxicity were Hg2+ (20 microgram/ml), Cu2+ (21 microgram/ml), Cr6+ (70 microgram/ml), Se4+ (73 microgram/ml), Ni2+ (160 microgram/ml), Cd2+ (175 microgram/ml), As3+ (304 microgram/ml) and As5+ (1610 microgram/ml). The elements tested that were either weak or noninhibitory at concentrations greater than 400 microgram/ml included Zn2+, Cr2+, Fe2+, Mn2+, Pb2+, and Co2+. Methylmercury was as inhibitory as mercuric chloride to the fermentation. When the inhibitory effect of Cd2+ was tested on separated bacterial and protozoal fractions, it was more inhibitory to the bacteria. The inhibitory effects of trace elements were also determined for a number of axenic cultures of rumen bacteria. The bacteria which most frequently exhibited the greatest sensitivity were Bacteroides succinogenses, Ruminococcus albus, Bacteroides amylophilus, and Eubacterium ruminantium. Those often exhibiting intermediate sensitivities included Butyrivibrio fibrisolvens, Selenomonas ruminantium, and Megasphera elsdenii, while Streptococcus bovis was very refractory to all elements tested. Rumen fluid provided a modest protective effect for the bacteria.     

Toxic effects exerted on methanogenic, nitrifying and denitrifying bacteria by chemicals used in a milk analysis laboratory

The toxic effects caused by the chemicals contained in wastewaters generated by laboratories involved in raw milk analyses were assessed using batch assays. These assays were carried out separately with methanogenic, ammonium-oxidizing, nitrite-oxidizing and denitrifying bacteria. Since sodium azide is one of the main components of the chemical mixture present in these streams, a set of assays was carried out with the whole chemical mixture, and another one was performed only with azide as the sole toxicant. The concentrations of all chemicals in the raw wastewaters (100%) were the fundamental references used to assess the relative concentrations corresponding to a decrease of 50% in bacterial activity (IC₅₀). The results obtained showed that nitrite-oxidizing bacteria were the most sensitive microorganisms, with IC₅₀ relative concentrations around 0.04%. The values obtained for the other groups were: 20, 20 and 33% for methanogenic, ammonium-oxidizing and denitrifying bacteria, respectively.     

Synthesis, characterization, and antibacterial activities of novel methacrylate polymers containing norfloxacin

A novel methacrylate monomer containing a quinolone moiety was synthesized and homopolymerized in N,N-dimethylformamide (DMF) by using azobisisobutyronitrile (AIBN) as an initiator. The new monomer was copolymerized with poly(ethylene glycol) methyl ether methacrylate (MPEGMA) in DMF using the same initiator. The monomer, homopolymer, and copolymer were characterized by elemental analysis, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), size exclusion chromatography (SEC), FTIR, (13)C NMR, and (1)H NMR. The antibacterial activities of the monomer as well as polymers were investigated against Staphylococcus aureus and Escherichia coli, which are representative of Gram-positive and Gram-negative bacteria, respectively. All compounds showed excellent antibacterial activities against these two types of bacteria. The antibacterial activities were determined using the shaking flask method, where 25 mg/mL concentrations of each compound were tested against 10(5) CFU/mL bacteria solutions. The number of viable bacteria was calculated by using the spread plate method, where 100 microL of the incubated antibacterial agent in bacteria solutions were spread on agar plates and the number of viable bacteria was counted after 24 h of incubation period at 37 degrees C.     

Meteorological factors and ambient bacterial levels in a subtropical urban environment

We conducted a study to investigate the characteristics and determinants of ambient bacteria in Taipei, Taiwan from August 2004 to March 2005. We monitored ambient culturable bacteria in Shin-Jhuang City, an urban area in the Taipei metropolitan areas, using duplicate Burkard Portable Air Samplers with R2A agar. The average concentration of total bacteria was 1,986 colony-forming units per cubic meter of air (CFU/m³) (median?=?780 CFU/m³) over the study period, with the highest level in autumn. Most bacterial taxa had similar seasonal variation, with higher concentrations in autumn and winter. During the study period, Gram negative rods and cocci were predominant. Multivariate analyses indicated that wind speed and wind direction significantly influenced ambient bacterial distribution. Temperature and relative humidity were also important environmental factors positively associated with ambient bacterial concentrations. We observed statistically significant relationships between ambient bacteria and air pollutants, including sulfur dioxide (SO₂), ozone, particulate matter (aerodynamic diameter ≤10 μm (PM₁₀)), methane and total hydrocarbons. The concentrations of methane and total hydrocarbons during the previous day were positively associated with total bacteria and Gram negative rods, respectively. Ozone level on the previous day had a negative relationship with Gram negative cocci. SO₂ level with a 3-day lag was positively correlated with concentrations of both total bacteria and Gram negative cocci. In the future, more longitudinal studies are needed to confirm the relationships and possible mechanisms between ambient bacteria and meteorological factors, as well as to evaluate the ecological and health impacts of ambient bacteria.     

The Toll-like receptors TLR2 and TLR4 do not affect the intestinal microbiota composition in mice

The interaction between intestinal epithelial cells and microbes is partly mediated by Toll-like receptors (TLRs). Sensing of Gram-positive and Gram-negative bacteria by TLR2 and TLR4, respectively, can result in immune system activation and in an exclusion of bacteria from the intestine. To test the impact of these TLRs on bacterial composition, germ-free TLR2/TLR4 double-knock out mice and the corresponding C57BL/10ScSn wild-type mice where associated with fecal bacteria from one single donor mouse. In addition, C3H/HeOuJ and BALB/c mice were used in this study. Fecal bacteria were monitored over 13 weeks with denaturing-gradient gel electrophoresis (DGGE). Colonic bacteria were enumerated by fluorescent in situ hybridization (FISH) and short-chain fatty acids (SCFA) were measured in caecal samples. No effect of the TLRs on intestinal microbiota composition and SCFA concentrations was observed. However, the microbiota composition as reflected by DGGE band patterns differed between C3H and BALB/c mice on the one hand and C57BL/10 mice on the other hand. Corresponding differences between the mouse strains were also observed in cecal propionic, valeric and i-valeric acid concentrations. No differences between the animals were observed in the numbers of bacteria detected by FISH. We conclude that genetic traits but not TLR2 and TLR4 have an impact on the intestinal microbiota composition.     

Growth characteristics of low-nutrient bacteria from the north-east and central Pacific Ocean

Abstract Low-nutrient, or oligotrophic, marine bacteria, defined operationally as those which grow well in the dilute nutrient concentrations typical of natural seawaters, were found in waters of the eastern and central Pacific Ocean. All isolates tested grew in sterile unsuplemented seawater, ultraviolet-irradiated seawater, artificial seawater and charcoal-treated artificial seawater. Under optimal conditions, doubling times for low-nutrient bacteria were generally 2–5 h. Low-nutrient bacteria are heterotrophs responding to added organic substrates including glucose, proline, acetate, peptone, algal extract and yeast extract. Organic substrate concentrations ( K _s values) necessary to yield half the maximum growth rates of low-nutrient bacteria ranged from 0.8–12 μg · 1⁻¹ (0.2–4.9 μg C · 1⁻¹). Bacterial inoculum size had little effect on doubling times, whereas low oxygen tensions and low temperatures reduced growth rates. Surfaces (glass beads) increased growth rates and maximum cell densities. The results of these laboratory studies indicate that low-nutrient bacteria play a major role in marine heterotrophic transformations of dissolved organic matter.     

Attached and Free Bacteria in an Estuary with Abundant Suspended Solids

Concentrations and glucose mineralization potentials ( V _max) of attached and free bacteria in waters from the Humber estuary were determined. The waters had high concentrations of suspended solids. Most bacteria were attached to suspended solids and V _max of attached bacteria was greater than that of free bacteria. These observations suggest that breakdown of organic matter is carried out mainly by attached bacteria. Examination of samples from other estuaries which contained less suspended solids than the Humber suggested that in these estuaries free bacteria might be more abundant than attached bacteria.     

The effects of agar concentration on the growth and morphology of submerged colonies of motile and non-motile bacteria

The growth and morphology of submerged bacterial colonies was investigated. Five separate colonial forms were recognized depending both on species and on agar concentration. These were (i) branched, dendritic structures seen only with Bacillus cereus ; (ii) lenticular colonies for all other species at high agar concentrations; (iii) small lobed to spherical colonies for non-motile organisms at low agar concentrations; (iv) and (v) large diffuse spherical colonies which can be further subdivided into 'snowball' or 'wispy' types for motile bacteria growing at agar concentrations below about 0·65% w/v. Viable count determinations suggested that agar concentration had little effect in the early stages of growth but that motile cells at low agar concentrations achieved higher cell numbers than did those in concentrations greater than 0·65% w/v. Transmission electron microscopy indicated that bacteria in lenticular colonies were tightly packed within lens-shaped splits in the agar whilst at low agar concentrations motile cells were well separated and appeared to move through the agar matrix.     

Strong Synergy between a Eukaryotic Antimicrobial Peptide and Bacteriocins from Lactic Acid Bacteria

The antimicrobial effect obtained upon combining the prokaryotic antimicrobial peptides (AMPs; more commonly referred to as bacteriocins) pediocin PA-1, sakacin P, and curvacin A (all produced by lactic acid bacteria [LAB]) with the eukaryotic AMP pleurocidin (from fish) has been investigated. The three LAB AMPs alone were active against gram-positive Listeria ivanovii bacteria at nanomolar concentrations, whereas they were inactive against gram-negative Escherichia coli bacteria. Pleurocidin alone was active against both of these types of bacteria at micromolar concentrations. Little if any synergy between the LAB AMPs and pleurocidin against the gram-positive L. ivanovii strain was obtained. In contrast, the LAB AMPs and pleurocidin acted highly synergistically against the gram-negative E. coli strain. Nanomolar concentrations of LAB AMPs increased the growth inhibitory potency of pleurocidin by about fourfold. When micromolar concentrations of LAB AMPs were combined with 2 μg of pleurocidin/ml, 100% growth inhibition was attained, whereas pleurocidin alone at a concentration of 2 μg/ml gave no growth inhibition. Most noteworthy, when high concentrations (128 μg/ml) of pleurocidin in the absence of LAB AMPs were used over a long period of incubation (1 week), some growth of E. coli was observed, whereas 16 μg of pleurocidin/ml completely abolished growth in the presence of 64 to 128 ng of LAB AMPs/ml over the same period of time. The results clearly demonstrate that combining eukaryotic and prokaryotic AMPs can greatly increase the specific activity and broaden the target-cell range of these peptides.     

Budding and prosthecate bacteria from freshwater habitats of various trophic states

Budding and prosthecate bacteria were enumerated in spring and summer by viable counting procedures in several freshwater habitats in Australia including oligotrophic lakes, a mesotrophic lake, and eutrophic ponds.Caulobacter spp. were the most numerous type encountered. They were present in the highest concentrations (exceeding 1000/ml) in the mesotrophic lake during the summer. Their proportion to total viable heterotrophic bacteria was also highest (35.1 to 37.7) in this habitat. From 17 to 330/mlCaulobacter spp. were counted in the eutrophic habitats where their proportion to total viable numbers was less than 1.0%. In the oligotrophic lakes they varied from 5 to 23/ml and comprised greater than 5% of the total viable count.Hyphomicrobium- like bacteria were also numerous in the mesotrophic lake and in one oligotrophic lake during the summer sampling period.Ancalomicrobium spp. occurred in high concentrations (130/ml) in the mesotrophic lake. Budding bacteria of thePlanctomyces-Pasteuria group were most numerous in the eutrophic habitats where as many as 240/ml were counted; their proportion to total heterotrophs remained relatively constant regardless of trophic state, however. A similar pattern was observed withProsthecobacter spp.     

Biotransformation of phosphogypsum in media containing different forms of nitrogen

Studies on the biotransformation of phosphogypsum (a waste product formed in the course of the production of phosphorous fertilizers) with the use of sulfate reducing bacteria (SRB) demonstrated that it is a good source of sulfates and biogenic elements for these bacteria, though the addition of organic carbon and nitrogen is necessary. The aim of this study was to investigate the form of nitrogen and C:N ratio in the medium on the growth of SRB community in cultures containing phosphogypsum. Batch community cultures of sulfate reducing bacteria were maintained in medium with phosphogypsum (5.0 g/l), different concentrations of sodium lactate (1.6 - 9.4 g/l) and different forms (NH4CI, CO(NH2)2, KNO3) and concentrations (0 - 250 mg/l) of nitrogen. The growth of SRB was studied in the C:N ratio of from 2:1 to 300:1. It was found that: 1 - the best source of nitrogen for SRB is urea, followed by ammonium, the worst were nitrates; 2 - the bacteria were also able to grow in medium without nitrogen but their activity was then by approximately 15% lower than in optimal growth conditions; 3 - in medium with KNO3 inhibition of sulfate reduction by approx. 50% was observed; 4 - the highest reduction of nitrates (removal of nitrate) in media with phosphogypsum and nitrates was at limiting concentrations of sodium lactate. This is probably caused by the selection under these conditions (low concentration of hydrogen sulfide) of denitrifying bacteria or sulfate reducing bacteria capable of using nitrates as an electron acceptor.