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Bacterial response to siderophore and quorum-sensing chemical signals in the seawater microbial community.

BACKGROUND: Oceans are iron-deficient and nutrient-poor environments. These conditions impart limitations on our understanding of and our ability to identify microorganisms from the marine environment. However, less of knowledge on the influence of siderophores and N-acyl homoserinelactone as interspecies communication signals on the bacterial diversity of seawater has been understood. RESULTS: In the presence of 0.1 nM of the commercial siderophore desferroixamine and the known quorum-sensing chemical signals, synthetic N-(3-oxo)-hexanoylhomoserine lactone (0.1 nM) or N-octanoylhomoserine lactone (0.1 nM), the total numbers of bacteria in S9905 seawater increased nearly three-fold, and nearly eight-fold in S0011 seawater as determined by DAPI staining and counting, and increased three-fold by counting colony forming units in S9905 seawater after 7 days of incubation. Similar bacterial changes in bacterial abundance were observed when high concentration of desferroixamine (1 microM) and each of homoserine lactone compounds (1 microM) were presented in seawater samples. The number of cultivable bacterial species observed was also found to increase from 3 (without addition) to 8 (with additions) including three unknown species which were identified by phylogenetic analysis of 16S rDNA sequences. The growth of unknown species was found to be related to their siderophore production with response to the addition of desferroixamine and N-acyl homoserine lactones under iron-limited conditions. CONCLUSION: Artificial addition of siderophores and HSLs may be a possible method to aid in the identification and isolation of marine bacterial species which are thought to be unknown.

4-Butyrolactone↗

Destruction of Listeria monocytogenes in sturgeon (Acipenser transmontanus) caviar by a combination of nisin with chemical antimicrobials or moderate heat.

The objective of this study was to investigate the effect of nisin in combination with heat or antimicrobial chemical treatments (such as lactic acid, chlorous acid, and sodium hypochlorite) on the inhibition of Listeria monocytogenes and total mesophiles in sturgeon (Acipenser transmontanus) caviar. The effects of nisin (250, 500, 750, and 1,000 IU/ml), lactic acid (1, 2, and 3%), chlorous acid (134 and 268 ppm), sodium hypochlorite (150 and 300 ppm), and heat at 60 degrees C for 3 min were evaluated for a five-strain mixture of L. monocytogenes and total mesophiles in sturgeon caviar containing 3.5% salt. Selected combinations of these antimicrobial treatments were also tested. Injured and viable L. monocytogenes cells were recovered using an overlay method. Treating caviar with > or =500 IU/ml nisin initially reduced L. monocytogenes by 2 to 2.5 log units. Chlorous acid (268 ppm) reduced L. monocytogenes from 7.7 log units to undetectable (<0.48 log units) after 4 days of storage at 4 degrees C. However, there were no synergistic effects observed for combinations of nisin (500 or 750 IU/ml) plus either lactic acid or chlorous acid. Lactic acid caused a slight reduction (approximately 1 log unit) in the microbial load during a 6-day period at 4 degrees C. Sodium hypochlorite was ineffective at the levels tested. Mild heating (60 degrees C for 3 min) with nisin synergistically reduced viable counts of L. monocytogenes and total mesophiles. No L. monocytogenes cells (<0.48 log units) were recovered from caviar treated with heat and nisin (750 IU/ml) after a storage period of 28 days at 4 degrees C.

Animals↗

Microbial population dynamics during fed-batch operation of commercially available garbage composters.

Microbial populations in terms of quantity, quality, and activity were monitored during 2 months of start-up operation of commercially available composters for fed-batch treatment of household biowaste. All the reactors, operated at a waste-loading rate of 0.7 kg day(-1) (wet wt), showed a mass reduction efficiency of 88-93%. The core temperature in the reactors fluctuated between 31 degrees C and 58 degrees C due to self-heating. The pH declined during the early stage of operation and steadied at pH 7.4-9.3 during the fully acclimated stage. The moisture content was 48-63% early in the process and 30-40% at the steady state. Both direct total counts and plate counts of bacteria increased via two phases (designated phases I, II) and reached an order of magnitude of 10(11) cells g(-1) (dry wt) at the steady state. Microbial community changes during the start-up period were studied by culture-independent quinone profiling and denatured gradient gel electrophoresis (DGGE) of PCR-amplified 16S rDNA. In all the reactors, ubiquinones predominated during phase I, whereas partially saturated menaquinones became predominant during phase II. This suggested that there was a drastic population shift from ubiquinone-containing Proteobacteria to Actinobacteria during the start-up period. The DGGE analysis of the bacterial community in one of the reactors also demonstrated a drastic population shift during phase I and the predominance of members of the phyla Proteobacteria and Bacteroidetes during the overall period. But this molecular analysis failed to detect actinobacterial clones from the reactor at any stage.

Actinobacteria↗

Shake-flask test for determination of biodegradation rates of (14)C-labeled chemicals at low concentrations in surface water systems.

A simple shake-flask surface water biodegradability die away test with (14)C-labeled chemicals added to microgram per liter concentrations (usually 1-100 microg/L) is described and evaluated. The aim was to provide information on biodegradation behavior and kinetic rates at environmental (low) concentrations in surface water systems. The basic principle of measurement was to determine evolved CO(2) indirectly from measurements of total organic activity in subsamples after stripping off their content of CO(2). Used with surface water alone the test simulates a pelagic environment and amended with sediments (0.1-1 dry weight/L) the test is intended to simulate a water environment with suspended solids (e.g., resuspended sediments). A protocol of the test used with the (14)C technique or with specific chemical analysis was recently developed by the International Organization for Standardization. Practical experience with the method is presented for a set of reference substances. These substances could be ranked in five groups of decreasing biodegradability: aniline>p-nitrophenol, 2, 4-dichlorophenoxyacetic acid>4-chloroaniline>maleic hydrazide, pentachlorophenol>atrazine. It was found that degradation rates and lag periods varied considerably among sampling sites and sometimes also among samples from the same site. No significant correlation could be established between degradation rates and microbial biomass estimates. Even small portions of added sediments greatly enhanced biodegradation of the absorbable compound pentachlorophenol, probably by providing sites for microbial attachment. Repeated tests indicated consistent degradation behavior for the readily degradable substances, whereas degradation sometimes stopped or failed with the more recalcitrant substances. A preadaptation step involving regular reinoculation with freshly collected surface water could, however, overcome the problems of false-negative results.

2,4-Dichlorophenoxyacetic Acid↗

Dissolved organic matter concentration and quality influences upon structure and function of freshwater microbial communities.

Past studies have suggested that the concentration and quality of dissolved organic matter (DOM) may influence microbial community structure. In this study, we cross-inoculated the bacterial communities from two streams and a dystrophic lake that varied in DOM concentration and chemistry, to yield nine fully crossed treatments. We measured dissolved organic carbon (DOC) concentration and heterotrophic microbial community productivity throughout a 72-h incubation period, characterized DOM quality by molecular weight, and determined microbial community structure at the initial and final time points. Our results indicate that all bacterial inoculate sources had similar effects upon DOC concentration and DOM quality, regardless of the DOM source. These effects included an overall decrease in DOM M (W) and an initial period of DOC concentration variability between 0-24h. In contrast, microbial communities and their metabolic rates converged to profiles that reflected the DOM source upon which they were growing, regardless of the initial bacterial inoculation. The one exception was that the bacterial community from the low-concentration and low-molecular-weight DOM source exhibited a greater denaturing gradient gel electrophoresis (DGGE) band richness when grown in its own DOM source than when grown in the highest concentration and molecular weight DOM source. This treatment also exhibited a higher rate of productivity. In general, our data suggest that microbial communities are selected by the DOM sources to which they are exposed. A microbial community will utilize the low-molecular-weight (or labile) DOM sources as well as parts of the high-molecular-weight (refractory) DOM, until a community develops that can efficiently metabolize the more abundant high-molecular-weight source. This experiment examines some of the complex interactions between microbial community selection and the combined factors of DOM quality and concentration. Our data suggest that the roles of aerobic aquatic heterotrophic bacteria in carbon cycling, as well as the importance of high-molecular-weight DOM as a carbon source, may be more complex than is conventionally recognized.

Bacteria↗

Evaluation of nitrogenous substrates such as peptones from fish:a new method based on Gompertz modeling of microbial growth.

Fish peptones from tuna, cod, salmon, and unspecified fish were compared with a casein one by using a new method based on Gompertz modeling of microbial growth. Cumulative results obtained from six species of bacteria, yeasts, and fungi showed that, in most cases, these fish peptones are very effective. Nevertheless, this study raised some questions about the standardization of fish raw material, the enzymatic hydrolysis of fish proteins, and the composition of the culture medium used for testing the peptones.

Animals↗

Development of autonomous control in a closed microbial bioreactor.

Space-based life support systems which include ecological components will rely on sophisticated hardware and software to monitor and control key system parameters. Autonomous closed artificial ecosystems are useful for research in numerous fields. We are developing a bioreactor designed to study both microbe-environment interactions and autonomous control systems. Currently we are investigating N-cycling and N-mass balance in closed microbial systems. The design features of the system involve real-time monitoring of physical parameters (e.g. temperature, light), growth solution composition (e.g. pH, NOx, CO2), cell density and the status of important hardware components. Control of key system parameters is achieved by incorporation of artificial intelligence software tools that permit autonomous decision-making by the instrument. These developments provide a valuable research tool for terrestrial microbial ecology, as well as a testbed for implementation of artificial intelligence concepts. Autonomous instrumentation will be necessary for robust operation of space-based life support systems, and for use on robotic spacecraft. Sample data acquired from the system, important features of software components, and potential applications for terrestrial and space research will be presented.

Artificial Intelligence↗

Description of heterotrophic bacteria occurring in paper mills and paper products.

AIMS: To isolate aerobic mesophilic bacilli and thermophilic bacteria from different paper mill samples and to evaluate their potential harmfulness. METHODS AND RESULTS: A total of 109 mesophilic and 68 thermophilic isolates were purified and characterized by automated ribotyping and partial 16S rDNA sequencing. The mesophilic isolates belonged to the genera Bacillus (13 taxa), Brevibacillus (three taxa) and Paenibacillus (five taxa). The thermophilic bacteria represented seven taxa of Bacillus, Geobacillus or Paenibacillus, four of proteobacteria and one of actinobacteria. The most frequently occurring bacteria were Bacillus cereus, B. licheniformis, Pseudoxanthomonas taiwanensis and bacteria closely related to Paenibacillus stellifer, P. turicensis or Leptothrix sp. One mill was contaminated throughout with bacteria of a novel mesophilic genus most closely related to Brevibacillus centrosporus and another with bacteria of a novel thermophilic genus most closely related to Hydrogenophilus thermoluteolus. One B. cereus isolate producing haemolytic diarrhoeal enterotoxin was detected and all the tested B. licheniformis isolates produced a metabolite toxic to boar sperm cells. CONCLUSIONS: The bacilli and thermophilic bacteria isolated represent species which should not present occupational hazards in paper mill environments. The most harmful bacterium detected was B. licheniformis and potentially also B. cereus. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge of the microbial diversity in a paper mill provides a rational basis for development of an effective controlling programme. A database constructed from the fingerprints generated using automated ribotyping helps to identify and trace the contamination routes of bacteria occurring in paper mills.

Bacillus↗

Low temperature bioremediation of oil-contaminated soil using biostimulation and bioaugmentation with a Pseudomonas sp. from maritime Antarctica.

AIMS: To identify native Antarctic bacteria capable of oil degradation at low temperatures. METHODS AND RESULTS: Oil contaminated and pristine soils from Signy Island (South Orkney Islands, Antarctica) were examined for bacteria capable of oil degradation at low temperatures. Of the 300 isolates cultured, Pseudomonas strain ST41 grew on the widest range of hydrocarbons at 4 degrees C. ST41 was used in microcosm studies of low temperature bioremediation of oil-contaminated soils. Microcosm experiments showed that at 4 degrees C the levels of oil degradation increased, relative to the controls, with (i) the addition of ST41 to the existing soil microbial population (bioaugmentation), (ii) the addition of nutrients (biostimulation) and to the greatest extent with (iii) a combination of both treatments (bioaugmentation and biostimulation). Addition of water to oil contaminated soil (hydration) also enhanced oil degradation, although less than the other treatments. Analysis of the dominant species in the microcosms after 12 weeks, using temporal temperature gradient gel electrophoresis, showed Pseudomonas species to be the dominant soil bacteria in both bioaugmented and biostimulated microcosms. CONCLUSIONS: Addition of water and nutrients may enhance oil degradation through the biostimulation of indigenous oil-degrading microbial populations within the soil. However, bioaugmentation with Antarctic bacteria capable of efficient low temperature hydrocarbon degradation may enhance the rate of bioremediation if applied soon after the spill. SIGNIFICANCE AND IMPACT OF THE STUDY: In the future, native soil bacteria could be of use in bioremediation technologies in Antarctica.

Alkanes↗

Occurrence of enteric rods, staphylococci and Candida in subgingival samples.

The frequency and percentage of enteric rods, staphylococci and Candida were determined in 973 subgingival samples collected from 535 patients subjected to different periodontal treatment procedures. The analysis was performed with culture technique using selective and nonselective media. One or more organisms were detected in 65.5% of the samples and in 76.7% of the patients. In most samples enteric rods, staphylococci and/or Candida constituted a small amount of the total microbial viable count. Enteric rods exceeded 10% of the total viable count in 30 samples. Staphylococci occurred in more than 10% in only 3 samples. In these 3 samples, enterics constituted more than 10% of the total viable count. Candida was not found to exceed 10% of the total viable count in any of the samples. No statistically significant correlation was found between the presence of any of the target microorganisms and kind of periodontal treatment procedure received, antibiotic administration or sample transport time.

Adolescent↗

[The microbial status of the periodontal pocket].

Analysis of components helped reveal the true relationships between the counts of individual representatives of the bacterial association in the contents of 18 periodontal pouches. Though the authors did their best to select a group of patients with the periodontitis course as similar as possible, the multidimensional grouping method has demonstrated the heterogeneity of this group. Four types of periodontal pouches were distinguished, differing by their bacterial status; this probably requires a differentiated approach to its correction.

Adult↗

Investigation of the hygienic safety of continuous positive airways pressure devices after reprocessing.

With the widespread use of continuous positive airways pressure (CPAP) therapy, the safety of CPAP devices after reprocessing is the subject of debate. In this study, the contamination of CPAP devices and the effectiveness of disinfection was investigated. A total of 122 CPAP devices were examined including 50 CPAP devices used by patients, which were examined before and after reprocessing. Seventy-two new CPAP devices that had not been in contact with patients served as controls. The results of this study show that the microbial contamination of new and used CPAP devices is only minimal. Contaminating micro-organisms were predominantly micro-organisms reflecting the normal environmental microflora such as Penicillium spp., Aspergillus spp., Micrococcaceae and Bacillaceae. Gram-negative species could only be found in rare cases. The data obtained give no indication of poor disinfection of CPAP devices.

Air Microbiology↗

Microbiologic spectrum and susceptibility of isolates: part II. Posttraumatic endophthalmitis. Endophthalmitis Research Group.

PURPOSE: To present the microbial spectrum and susceptibilities of isolates in posttraumatic endophthalmitis. METHOD: Isolates from 182 eyes of 182 patients who underwent vitrectomy for posttraumatic endophthalmitis were examined. RESULTS: One hundred thirteen (62.1%) of 182 vitreous samples were culture-positive, and 23 (20.4%) of 113 culture-positive cases were polymicrobial, including three (2.7%) trimicrobial cases, yielding a total of 139 isolates. Isolates included 63 (45.3%) gram-positive cocci, 24 (17.3%) gram-positive bacilli, 25 (18.0%) gram-negative organisms, seven (5.0%) Actinomycetes-related organisms, and 20 (14.4%) fungi. Susceptibilities to amikacin, ceftazidime, chloramphenicol, cefazolin, ciprofloxacin, gentamicin, and vancomycin are reported. CONCLUSIONS: This study represents a large series on microbial spectrum and susceptibilities in posttraumatic endophthalmitis. We report a high prevalence of gram-positive bacilli species and polymicrobial infections containing gram-negative species, underscoring the importance of broad-spectrum, combination antibiotics in the empiric treatment of posttraumatic endophthalmitis.

Anti-Bacterial Agents↗

Microbial community dynamics and evaluation of bioremediation strategies in oil-impacted salt marsh sediment microcosms.

Microbial community dynamics in wetlands microcosms emended with commercial products (surfactant, a biological agent, and nutrients) designed to enhance bioremediation was followed for 3 months. The effectiveness of enhanced degradation was assessed by determining residual concentrations of individual petroleum hydrocarbons by GC/MS. The size and composition of the sediment microbial community was assessed using a variety of indices, including bacterial plate counts, MPNs, and DNA hybridizations with domain- and group-specific oligonucleotide probes. The addition of inorganic nutrients was the most effective treatment for the enhancement of oil degradation, resulting in marked degradation of petroleum alkanes and a lesser extent of degradation of aromatic oil constituents. The enhanced degradation was associated with increases in the amount of extractable microbial DNA and Streptomyces in the sediment, although not with increased viable counts (plate counts, MPN). Bacteria introduced with one of the proprietary products were still detected in the microcosms after 3 months, but were not a major quantitative constituent of the community. The biological product enhanced oil degradation relative to the control, but to a lesser extent than the nutrient additions alone. In contrast, application of the surfactant to the oil-impacted sediment decreased oil degradation.

Biodegradation, Environmental↗

Eugenol stimulates lactate accumulation yet inhibits volatile fatty acid production and eliminates coliform bacteria in cattle and swine waste.

AIM: To determine how eugenol affects fermentation parameters and faecal coliforms in cattle and swine waste slurries stored anaerobically. METHODS AND RESULTS: Waste slurries (faeces:urine:water, 50:35:15) were blended with and without additives and aliquoted to triplicate 1-l flasks. Faecal coliforms were eliminated in cattle and swine waste slurries within 1 or 2 days with additions of eugenol at 10.05 mm (0.15%) and 16.75 mm (0.25%). At these concentrations volatile fatty acids (VFA) were reduced ca 70 and 50% in cattle and swine waste, respectively, over 6-8 weeks. Additionally, in cattle waste, eugenol stimulated the accumulation of lactate (>180 mm) when compared with thymol treatment (20 mm lactate). In swine waste, lactate accumulation did not occur without additives; eugenol and thymol stimulated lactate accumulation to concentrations of 22 and 32 mm, respectively. CONCLUSIONS: Eugenol added to cattle waste may be more beneficial than thymol because not only does it effectively control faecal coliforms and odour (VFA production), it also stimulates lactate accumulation. This in turn, causes the pH to drop more rapidly, further inhibiting microbial activity and nutrient emissions. SIGNIFICANCE AND IMPACT OF THE STUDY: Plant essential oils have the potential to solve some of the environmental problems associated with consolidated animal feeding operations. Thymol and eugenol reduce fermentative activity, thus, have the potential to reduce emissions of greenhouse gases and odour, and curtail transmission of pathogens in cattle and swine wastes.

Animals↗

Effects of xylitol, xylitol-sorbitol, and placebo chewing gums on the plaque of habitual xylitol consumers.

Xylitol reduces plaque but the reduction mechanism is largely unknown. The main aim of the present study was to determine whether the xylitol-induced reduction in the amount of plaque and the number of mutans streptococci could be demonstrated in subjects with (presumably) high levels of xylitol-resistant (XR; not inhibited by xylitol) mutans streptococci acquired following previous xylitol consumptions. 37 healthy dental students participated in the double-blind study. All subjects had been uncontrolled, habitual consumers of xylitol-containing products for at least 1 yr before the study. A 1-month washout period was followed by a 2-week test period during which either xylitol, xylitol-sorbitol or unsweetened chewing gum base was chewed 3-5 x a day. Plaque and saliva samples were collected at baseline and at the 2-week point for determination of the amount of plaque, microbiological variables, and hydrolytic enzymes. Mixtures of xylitol and sorbitol seemed to perform equally well with respect to reduction in the amount of plaque but not the number of mutans streptococci. Thus, polyols were the active ingredients of chewing gums able to modulate the amount of plaque and its microbial composition. Xylitol reduced plaque with a mechanism which appeared not to be associated with the study-induced changes in the proportion (%) of mutans streptococci in plaque, the number of salivary mutans streptococci, the proportion of XR strains in plaque or saliva, or the hydrolytic enzyme activities of plaque.

Adult↗

Microbiology of the oil fly, Helaeomyia petrolei.

Helaeomyia petrolei larvae isolated from the asphalt seeps of Rancho La Brea in Los Angeles, Calif., were examined for microbial gut contents. Standard counts on Luria-Bertani, MacConkey, and blood agar plates indicated ca. 2 x 10(5) heterotrophic bacteria per larva. The culturable bacteria represented 15 to 20% of the total population as determined by acridine orange staining. The gut itself contained large amounts of the oil, had no observable ceca, and maintained a slightly acidic pH of 6.3 to 6.5. Despite the ingestion of large amounts of potentially toxic asphalt by the larvae, their guts sustained the growth of 100 to 1,000 times more bacteria than did free oil. All of the bacteria isolated were nonsporeformers and gram negative. Fourteen isolates were chosen based on representative colony morphologies and were identified by using the Enterotube II and API 20E systems and fatty acid analysis. Of the 14 isolates, 9 were identified as Providencia rettgeri and 3 were likely Acinetobacter isolates. No evidence was found that the isolates grew on or derived nutrients from the asphalt itself or that they played an essential role in insect development. Regardless, any bacteria found in the oil fly larval gut are likely to exhibit pronounced solvent tolerance and may be a future source of industrially useful, solvent-tolerant enzymes.

Animals↗

Comparison of the microflora on organically and conventionally grown spring mix from a California processor.

Considerable speculation has occurred concerning the potential for higher numbers of foodborne pathogens on organically grown produce compared with produce not grown organically. The microflora composition of spring mix or mesclun, a mixture of multiple salad ingredients, grown either by organic or conventional means was determined. Unwashed or washed spring mix was obtained from a commercial California fresh-cut produce processor who does not use manure in their cultivation practices. Fifty-four samples of each type of product were supplied over a 4-month period. Analysis included enumeration of total mesophiles, psychrotrophs, coliforms, generic Escherichia coli, lactic acid bacteria, yeasts, and molds. In addition, spring mix was analyzed for the presence of Salmonella and Listeria monocytogenes. The mean populations of mesophilic and psychrotrophic bacteria, yeasts, molds, lactic acid bacteria, and coliforms on conventionally grown spring mix were not statistically different (P > 0.05) from respective mean populations on organically grown spring mix. The mean population of each microbial group was significantly higher on unwashed spring mix compared with the washed product. Of the 14 samples found to contain E. coli, eight were from nonwashed conventional spring mix, one was from washed conventional spring mix, and four were from nonwashed organic spring mix. Salmonella and L. monocytogenes were not detected in any of the samples analyzed.

Agriculture↗