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OBJECTIVE: To study the preparation of seeding type immobilized microorganisms and their degradation characteristics on di-n-butyl phthalate (DBP). METHODS: Diatomite, clinoptilolite, silk zeolite, and coal fly ash were chosen as reserved materials and modified. Their adsorption capacity and intensity in the bacteria were determined and the best carrier was picked out. The seeding type immobilized microorganisms were prepared by the best carrier and then it degraded DBP under different primary concentration, vibration rate, pH, temperature in the presence of metal compounds. RESULTS: The adsorption capacity of the modified coal fly ash, silk zeolite, clinoptilolite and zeolite was 44.2%, 71.6%, 84.0%, and 94.4%, respectively, which was 1.66, 1.49, 1.37, and 1.16 times as high as that of their natural state. Their adsorption intensity was 72.1%, 90.5%, 90.1%, and 91.1% in turn. The modified diatomite was selected to prepare the seeding type immobilized microorganisms. When the primary DBP concentration was 100 to 500 mg/L, the DBP-degraded rate of the immobilized microorganisms could be above 80%. The degradation activity of both the dissociative and immobilized microorganisms was higher in vibration than in stillness. When pH was 6.0 to 9.0, the DBP-degraded rate of the immobilized microorganisms was above 82%, which was higher than the dissociative microorganisms. When the temperature was between 20 degrees C and 40 degrees C, the DBP-degraded rate could reach 84.5% in 24 h. The metal compounds could inhibit the degradation activity of both the dissociative and immobilized microorganisms. The degradation process of the immobilized microorganisms could be described by the first-order model. CONCLUSION: The adsorption capacity of the diatomite, clinoptilolite, silk zeolite and coal fly ash on DBP-degrading bacteria can be improved obviously after they are modified. The modified diatomite is best in terms of its adsorption capacity and intensity. Its seeding type immobilized microorganisms could degrade DBP effectively and is more adaptable to DBP load, temperature, pH than the dissociative microorganisms. The metal compounds could inhibit the activity of both the immobilized and dissociative microorganisms. The degradation reaction of the immobilized microorganisms on DBP is consistent with the first-order model.
OBJECTIVE: To study the biodegradation characteristics of seeding type immobilized microorganism on dibutyl phthalate (DBP). METHODS: The immobilized microorganism was made to adsorb DBP degradation dominant bacteria by using modified diatomite as carrier, then it degraded DBP under different primary concentration, vibration rate, pH, temperature and at the presence of metal compounds. The degradation kinetics was analyzed. RESULTS: When DBP primary concentration was 100-500 mg/L, the adsorptive immobilized microorganism could maintain relatively high activity and the DBP degraded rate was above 80% in 24h. Dissociative and immobilized microorganism could get higher degradation activity in vibration than in stillness. When pH was 6.0 - 9.0, the degradation rate of immobilized microorganism on DBP was above 82% in 24h and its activity is higher than dissociative microorganism. In the range of 20 degrees C to 40 degrees C, the DBP degraded rate by immobilized microorganism could reach 84.5% in 24h. If mental compounds existed in the DBP water sample, the degradation activities of dissociative and immobilized microorganism were inhibited obviously. The form of DBP degradation kinetics could be described as the first-order model. CONCLUSION: The immobilized microorganisms using diatomite as carrier could degrade DBP effectively. The adsorptive immobilized microorganism was more adapted to DBP load, temperature and pH than dissociative microorganism. The mental compounds could inhibited their activities. The degradation reaction of adsorptive immobilized microorganisms on DBP was according with the first-order model.
The quantity of pharmaceutical and cosmetic products with different composition and the diversity of contaminating microorganisms set up special problems for each product with its specific field of application. It is not exact to generalize the hazard of contaminating microorganisms to product cathegories. Some product cathegories however are more imperilled to be contaminated with microorganisms and they are more hazardous than other ones. The most important component of the living matter is water that decides on growth, multiplication and survival of microorganisms in drugs and cosmetics. Products with a high content of available water are the most problematic ones. The consequences of survival and multiplication of microorganisms in relation to metabolic activity, pathogenicity and route of administration can be harmless or fatal. The nutritive wants of many microorganisms are very small, even simple salt solutions do not exclude microbial growth. There scarcely exist substrates not used as energy source by any microorganism and so it does not wonder that desinfectants are also used. Added preservatives may be absorbed or inactivated by suspended anorganic components, swelling agents, solubilizers etc., the containers or its closing. In the water phase of emulsions, in which under special conditions smaller amounts of the preservative are available, microorganisms sometimes can multiply. Skin care products neither belonging to pharmaceuticals nor to cosmetics present special clinical problems. Many times creams and lotions contain ingredients metabolized by microorganisms. Ointments and oils can be overgrown only by highly specialized organisms but if there is condensed water as a film at boundary surfaces much more microorganism species can grow. When using cosmetics it must be considered that parts of them will get into the eye. The eye is really endangered by Pseudomonads, which can according to the composition multiply in shampoos and whose contact to the eye can not be excluded. Syrups and solutions contain sugar concentrations, which suppress growth of most microorganisms except osmotolerant germs. Because of their low water content powders normally are protected against microbial growth but they should be free from clostridial spores for a mixture of powder and sweat is a good nutrient medium to Clostridia and fungi. Powders for surgical use must be sterile. Tablets and dragees do not allow microbial multiplication because of their low water activity. Under bad storage conditions a local increase of water content may permit microbial growth at the surface, especially in hygroscopicity. Fortunately there is seen a trend of not using multi-dose-containers for injections because of the high risk of contamination. It is to develop single-dose-containers also for other groups of preparations especially for eye-preparations. Many products microbiologically unobjectionable produced are contaminated in use, they contain the spectrum of microorganisms of the area of application...
In this study, a quinone profiling method was applied to clarify the differences in community structure between suspended and sessile microorganisms in rivers. The compositions of microbial quinone of 6 sites for 4 rivers were analyzed. Ubiquinone (UQ)-8, UQ-10, menaquinone (MK)-7, and plastoquinone (PQ)-9 were observed in all samples of suspended and sessile microorganisms for the sites investigated. The dominant quinone species in suspended microorganisms was ubiquinone, and that in sessile microorganism was photosynthetic quinones (namely PQ-9 and vitamin K1). This indicated that aerobic bacteria were abundant in the suspended microorganisms, and photosynthetic microorganisms such as micro-algae and cyanobacteria dominated in the sessile microorganisms. The quinone concentration in the river waters tested, which reflects the concentration of suspended microorganisms, ranged from 0.045 to 1.813 nmol/L. The microbial diversities of suspended and sessile microorganisms calculated based on the composition of all quinones were in the range from 3.4 to 7.5, which was lower than those for activated sludge and soils. Moreover, the diversity of heterotrophic bacteria for sessile microorganisms in the rivers was higher than that for the suspended microorganisms.
BACKGROUND: Successful application of growth inhibition techniques for quantitative determination of bacteriocins relies on the sensitivity of the applied indicator microorganism to the bacteriocin to which is exposed. However, information on indicator microorganisms' performance and comparisons in bacteriocin determination with bioassays is almost non-existing in the literature. The aim of the present work was to evaluate the parameter "indicator microorganism" in bioassays carried out on solid -agar diffusion assay- and liquid -turbidometric assay- substrates, applied in the quantification of the most studied bacteriocin nisin. RESULTS: The performance of characterized microorganisms of known sources, belonging to the genera of Lactobacillus, Pediococcus, Micrococcus and Leuconostoc, has been assessed in this work in the assays of plate agar diffusion and turbidometry. Dose responses and sensitivities were examined and compared over a range of assay variables in standard bacteriocin solutions, fermentation broth filtrates and processed food samples. Measurements on inhibition zones produced on agar plates were made by means of digital image analysis. The data produced were analyzed statistically using the ANOVA technique and pairwise comparisons tests. Sensitivity limits and linearity of responses to bacteriocin varied significantly among different test-microorganisms in both applied methods, the lower sensitivity limits depending on both the test-microorganism and the applied method. In both methods, however, only two of the nine tested microorganisms (Lactobacillus curvatus ATCC 51436 and Pediococcus acidilactici ATCC 25740) were sensitive to very low concentrations of the bacteriocin and produced a linear-type of response in all kinds of samples used in this work. In all cases, very low bacteriocin concentrations, e.g. 1 IU/ml nisin, were more accurately determined in the turbidometric assay. CONCLUSION: The present work shows that in growth inhibition techniques used in bacteriocin quantification, the choice of the indicator microorganism is critical. Evaluation of sensitivity levels and type of produced responses showed that they can vary widely among different test-microorganisms and different applied methods, indicating that not all microorganisms can be used successfully as indicators and that measurements of growth inhibition in liquid media produce more reliable results.
A simple biofilm model was developed to simulate the competition between two microorganisms for a common inhibitory substrate. The following assumptions were made for the simulations: (1) the biofilm has a uniform thickness and is composed of 5 segments, (2) growth of two microorganisms A and B which utilize the common substrate is expressed by the Haldane kinetics with a spatial limitation term and is independent of the other microorganism in the biofilm reactor, and (3) diffusion of the substrate, movement of the microorganisms, and continuous loss of the biomass by shearing are expressed by Fick's Law-type equations. The qualitative behavior of the biofilm reactor is characterized by five regions, I-V, depending on the operation conditions, the substrate concentration in feed, and the dilution rate. In region I, both microorganisms are washed out of the biofilm reactor. In region II, microorganism B is washed out, and in region III, microorganism A is washed out of the biofilm. In region IV, both microorganisms coexist with one another. In region V, both microorganisms coexist with a sustained oscillatory behavior. Convergence to regions I-V depends on the initial conditions. In regions II-V, washout of either or both microorganisms is also observed with initial conditions too far away.
Biofiltration is distinguished from other biological waste treatments by the fact that there is a separation between the microorganisms and the treated waste. In biofiltration systems the microorganisms are immobilized to the bedding material, while the treated fluid flows through it. In recent decades, a vast amount of literature has been written on single experiments involving the treatment of fluids by immobilized microorganisms. Several artificial immobilization methods have been examined and impressive results have been achieved in the treatment of fluids with one of the artificial immobilization methods the entrapment of microorganisms within polymer beads. This method, even though it needs to be improved, seems to have a future potential in commercial biofiltration systems. The methods of artificial immobilization of microorganisms within biofiltration systems have several advantages, but also suffer from several disadvantages in comparison to the treatment of fluids by naturally attached microorganisms. Understanding the mechanisms and forces responsible for the attachment of microbes to the bedding material, in attempt to improve this attachment, is of the utmost importance. Further improvement of the artificial entrapment of microorganisms within polymers will allow the exploitation of the advantages of this method in the treatment of fluids. The aim of this review essay is to introduce the main principles of two immobilization processes - the self-attachment of microorganisms to the bedding material and the artificial entrapment of microorganisms within polymer beads. Both treatments of liquids and gases with each immobilization process are discussed. The advantages and disadvantages of each immobilization process are pointed out and different aspects of the fluid treatment with the two immobilization processes are compared.
An improved data analysis method is described for rapid identification of intact microorganisms from MALDI-TOF-MS data. The method makes no use of mass spectral fingerprints. Instead, a microorganism database is automatically generated that contains biomarker masses derived from ribosomal protein sequences and a model of N-terminal Met loss. We quantitatively validate the method via a blind study that seeks to identify microorganisms with known ribosomal protein sequences. We also include in the database microorganisms with incompletely known sets of ribosomal proteins to test the specificity of the method. With an optimal MALDI protocol, and at the 95% confidence level, microorganisms represented in the database with 20 or more biomarkers (i.e., those with complete or nearly completely sequenced genomes) are correctly identified from their spectra 100% of the time, with no incorrect identifications. Microorganisms with seven or less biomarkers (i.e., incompletely sequenced genomes) are either not identified or misidentified. Robustness with respect to variations in sample preparation protocol and mass analysis protocol is demonstrated by collecting data with two different matrixes and under two different ion-mode configurations. Statistical analysis suggests that, even without further improvement, the method described here would successfully scale up to microorganism databases with roughly 1000 microorganisms. The results demonstrate that microorganism identification based on proteome data and modeling can perform as well as methods based on mass spectral fingerprinting.
A method is reported that combines the microscopic determinations of specific, individual, respiring microorganisms by the detection of electron transport system activity and the total number of organisms of an estuarine population by epifluorescence microscopy. An active cellular electron transport system specifically reduces 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride (INT) to INT-formazan, which is recognized as opaque intracellular deposits in microorganisms stained with acridine orange. In a comparison of previously described sample preparation techniques, a loss of >70% of the counts of INT-reducing microorganisms was shown to be due to the dissolution of INT-formazan deposits by immersion oil (used in microscopy). In addition, significantly fewer fluorescing microorganisms and INT-formazan deposits, both </=0.2 mum in size, were found for sample preparations that included a Nuclepore filter. Visual clarity was enhanced, and significantly greater direct counts and counts of INT-reducing microorganisms were recognized by transferring microorganisms from a filter to a gelatin film on a cover glass, followed by coating the sample with additional gelatin to produce a transparent matrix. With this method, the number of INT-reducing microorganisms determined for a Chesapeake Bay water sample was 2-to 10-fold greater than the number of respiring organisms reported previously for marine or freshwater samples. INT-reducing microorganisms constituted 61% of the total direct counts determined for a Chesapeake Bay water sample. This is the highest percentage of metabolically active microorganisms of any aquatic population reported using a method which determines both total counts and specific activity.
CONTEXT: The National Inventory of Clinical Laboratory Testing Services (NICLTS) was designed to give an unbiased estimate of all patient testing performed by laboratories registered under the Clinical Laboratory Improvement Amendments in 1996. OBJECTIVE: Survey data were used to develop a profile of laboratory testing primarily intended to identify microorganisms or antibodies to these microorganisms. DESIGN: Estimates of the extent of microorganism identification were derived from the NICLTS database by identifying associated tests and methods. The volumes for tests performed at locations that primarily prepared blood components for distribution were excluded. Organisms of public health importance were identified from the National Notifiable Disease list maintained by the Centers for Disease Control and Prevention. PARTICIPANTS: Laboratories that were enrolled in the 1996 Online Certification Survey and Reporting System, maintained by the US Health Care Finance Administration, and that performed laboratory testing in 1996. OUTCOME MEASURE: Estimated volumes and associated confidence limits by test, method, specimen type, public health importance, and testing location. RESULTS: Excluding testing of the blood supply, 315 million tests (95% confidence limits, 280-354 million tests) were performed in the United States for microorganism identification. Those tests for which public health consensus requires national reporting represented 38% of this total. Although hospitals performed 46% of all microorganism identification, they only performed 33% of the testing for microorganisms of public health importance. Independent and specialty laboratories performed 38% of all testing but 65% of the testing for microorganisms of public health importance. Direct methods (methods not involving culture) were used in 77% of the tests for microorganisms of public health importance and in 42% of all identification tests. CONCLUSIONS: The distribution of microorganism identification testing found using NICLTS data is consistent with plans to modernize the public health surveillance system in the United States.
Freezing of prokaryotic and eukaryotic microorganisms is the main interest in the study of cold stress responses of living organisms. In parallel, applications which arise from this approach are of two types: (i) optimization of the frozen starters used in food processing; and (ii) improvement of the ex situ preservation of microorganisms in collections. Currently, cryopreservation of microorganisms in collections is carried out in cryotubes, and bibliographical references related to freezing microorganisms packaged in straws are scarce. In this context, a preliminary study was completed to evaluate the technological potential of ionomeric resin straws compared to polycarbonate cryo-tubes. Survival under freezing stress was tested on three microorganisms selected for their biotechnological interest: two lactic acid bacteria, Lactococcus lactis subsp. cremoris and Lactobacillus delbrueckii subsp. bulgaricus and a deuteromycete fungus, Geotrichum candidum. The stress was carried out by repeated freezing-thawing cycles to artificially accelerate the lethal effect of freezing on the microorganisms. Two main results were obtained: (i) the survival rate values (per freezing-thawing cycle) seems to depend on the thermal type of the studied microorganism, and (ii) there was no, under our experimental conditions, significant difference between straws and tubes. However, conservation in the resin straws lead to a slight increase in the survival of L. cremoris and G. candidum compared to microtubes. In those conditions, straws seems an alternative system to securely store frozen microorganisms with three main characteristics: (i) a high resistance to thermal stress, (ii) a safe closing by hermetic weld, and (iii) a system for inviolable identification.
Activated sludge technology is more used than any other for biological treatment of wastewater. However, filamentous bulking is a very common problem in activated sludge plants, chlorine being the chemical agent normally used to control it. In this work the effect of chlorine on microorganisms present in activated sludge flocs was assessed by a respirometric technique (oxygen uptake rate, OUR) and by the INT-dehydrogenase activity test (DHA) measured by two techniques: spectrophotometry (DHA(a)) and image analysis (DHA(i)). Both DHA tests were optimized and correlated with the respirometric technique (OUR) using pure cultures of a filamentous microorganism (Sphaerotilus natans) under chlorine inhibition. Using these correlations the tested methods were applied to determine the action of chlorine on respiratory activity in activated sludge. The OUR and the DHA(a) quantifies the action of chlorine on the total respiratory activity (RA) of flocs (filamentous and floc-forming bacteria); in contrast, the DHA(i) test evaluates specific action of chlorine on the RA of filamentous microorganisms. In activated sludge flocs containing filamentous microorganisms, a chlorine dose of 4.75 mgCl(2) (gVSS)(-1) with a contact time of 20 min reduced about 80% of the RA of filamentous bacteria while affecting only 50-60% of the total RA of flocs. Besides, a chlorine dose of 7.9 mgCl(2) (gVSS)(-1) produced the total respiratory inactivation of filamentous microorganisms after 10 min contact, however, with this dose the total RA of activated sludge flocs was reduced only about 45-65%; controlling filamentous bulking without affecting too much floc-forming bacteria. At the tested chlorine concentrations the inhibition of filamentous microorganisms was higher than in the whole activated sludge. Although floc-forming microorganisms were demonstrated to be more susceptible to chlorine than filamentous in pure cultures, results obtained in the present work confirmed that it is the location of the filamentous microorganisms in the flocs and the presence of extracellular polymer substances which largely determines their higher susceptibility to chlorine; consequently this feature plays a critical role in bulking control.
A mathematical model concerning the interaction of plants and rhizospheric microorganisms on complete mineral medium and under nitrogen limitation has been constructed. The model takes into account the closeness of plants and microorganisms in terms of the matter released by the plant and consumed by the microorganisms. The effect of rhizospheric microorganisms on plant growth with normal carbon dioxide and complete mineral medium has been demonstrated. Plants interacting with microorganisms have a greater biomass than plants growing without microorganisms. Wheat growth stimulation by metabolites of rhizospheric microorganisms under laboratory conditions on artificial soil has been experimentally demonstrated (Pechurkin, 1997). Under nitrogen limitation, the biomass of plants, with or without microorganisms, is identical, and is substantially reduced as compared with the medium with standard nitrogen.
Mixed ruminal microorganisms were harvested from a lactating dairy cow and preserved frozen or lyophilized. Fermentation characteristics of freshly strained ruminal fluid, frozen microorganisms, or lyophilized microorganisms were evaluated during a 24-h pre-incubation and a 4-h incubation with test proteins. Differences observed during the first 4 to 6 h in total amino acid concentration, optical density, pH and VFA concentrations, acetate:propionate ratio, and lactate concentration largely disappeared later in the pre-incubation. Protein degradation rates determined for expeller and solvent meals were .015 and .092 h-1, .015 and .101 h-1, and .005 and .019 h-1, with fresh ruminal fluid, frozen microorganisms, and lyophilized microorganisms, respectively. Regression of degradation rates obtained with fresh ruminal fluid on those obtained with pre-incubated, frozen microorganisms indicated the two methods were well correlated (r2 = .98 and .94 in two experiments). Mean in vitro degradability obtained for 17 feeds using pre-incubated, frozen microbes was 89% of that obtained using the in situ method; however, in situ degradation rates for these same feeds averaged only 67% of those obtained using frozen microorganisms. Ruminal undegraded protein values for nine samples of heated soybeans and soybean meal, determined using frozen microbes, were overestimated relative to in vivo values (in vivo = 1.1 + .8 in vitro; r2 = .77). These results indicated that ruminal microorganisms can be preserved by freezing and used as the inoculum for in vitro determination of ruminal protein degradation after overnight pre-incubation.
Ruminal microorganisms, preserved either lyophilized or frozen, were compared with freshly strained ruminal fluid for proteolytic activity and as inoculum source for determination of ruminal protein degradation rates by the inhibitor in vitro method. Dialysis and glycerol addition had no effect on the proteolytic activity of preserved microorganisms. Net release of NH3 and total amino acids from protein using the fluid plus particle-associated microorganisms was higher than that found using the fluid-associated microorganisms alone. Method of inoculum preservation altered total proteolytic activity, but harvesting bacteria using centrifugal force greater than 5,000 x g did not increase proteolytic activity of the pellet. The proposed method for harvesting and preserving microorganisms consisted of centrifuging strained ruminal fluid at 5,000 x g (30 min at 4 degrees C), stirring the pellet in a 50:50 (vol/vol) solution of glycerol-McDougall's buffer for 15 min, and then storing at -20 degrees C. Protein degradation rates in incubations with preserved microorganisms were four to eight times slower than when using fresh ruminal fluid; however, feed proteins were ranked similarly for degradation rate. Preincubating the preserved microorganisms reduced blank concentrations of NH3 and total amino acid and increased protein degradative activity of the preserved inoculum. Degradation rates with preincubated, preserved inocula were similar to those obtained using fresh ruminal fluid. These results indicated that mixed ruminal microorganisms can be preserved by freezing and, after a preincubation period of 6 h, used as the inoculum source for in vitro estimation of ruminal protein degradation.
In a previous series of rocket- and satellite-borne experiments, microorganisms were exposed to space between altitudes of 60 to 460 km for periods between 3 minutes and 4 months. The results showed that some of the unprotected microorganisms survived the direct exposure for up to 17 hours. Complete survival was found when the microorganisms were shielded from nonpenetrating radiation. These results made it desirable to systematically study the lethal influence of the space environment on a wider spectrum of unprotected microorganisms over various exposure periods. This communication presents the results of exposure experiments on board the Gemini XII satellite. The microorganisms were dried on plastic coated aluminum plates using techniques which will be described elsewhere. During the flight of the Gemini XII space capsule, the microorganisms were directly exposed to space for approximately 6 1/2 hours. After the successful completion of the Gemini XII mission the payload was returned to the laboratory for elution and titration of the microorganisms using techniques described elsewhere. The results showed that again survival of some microorganisms had occurred. An attempt is made to integrate these results with data obtained in previously published experiments.