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Mixed Cultures (mixed + culture)
Selected AbstractsVIRULENCE RESPONSE OF A SALMONELLA TYPHIMURIUM HILA:LACZY FUSION STRAIN TO SPENT MEDIA FROM PURE CULTURES OF SELECTED BACTERIA AND POULTRY CECAL MIXED CULTUREJOURNAL OF FOOD SAFETY, Issue 3 2002J.D. NUTT ABSTRACT Virulence gene expression in Salmonella is triggered by a variety of environmental factors including changes in the gastrointestinal environment of birds during different dietary regimes. The objective of this study was to determine if growth of specific microorganisms alters the environmental conditions sufficiently to signal Salmonella Typhimurium virulence response. Spent media was obtained from a Salmonella Typhimurium hilA:lacZY fusion strain, a poultry Salmonella Typhimurium strain, Eschcrichia coli K12, and Lactobacillus caseii Spent media samples were collected after 2, 4, 8 and 24 h of growth in brain heart infusion broth (BHI) and M9 media, ,-galactosidase assays were performed on the samples to determine virulence expression. Virulence response to Salmonella, spent media was 2-fold greater than Lactobacillus spent media at 4, 8 and 24 h growth (P < 0.05). Virulence expression almost doubled when exposed to Salmonella Typhimurium (NONA) spent media compared to mixed culture spent media at 4 h, and Salmonella Typhimurium (NONA) was significantly higher than mixed culture spent media at 24 h (P < 0.05). Based on these results, it appears that growth of similar bacterial species may alter the composition of rich media sufficiently to influence virulence. [source] A Kinetic Model for Suspended and Attached Growth of a Defined Mixed CultureBIOTECHNOLOGY PROGRESS, Issue 3 2005Kawai Tam Kinetic experiments were carried out in a semicontinuous wastewater treatment process called self-cycling fermentation (SCF) using a defined mixed culture and various concentrations of synthetic brewery wastewater. The same consortium, which had been previously identified as Acinetobacter sp., Enterobacter sp., and Candida sp., were used in these experiments. The overall rate of substrate removal was attributable to both suspended microbes and the biofilm that formed during the treatment process. A rate expression was developed for the SCF system for a range of synthetic wastewaters containing glucose and various initial concentrations of ethanol and maltose. The data indicated that substrate removal by the suspended cells was directly related to the biomass concentration. However, substrate removal by the biofilm was apparently not affected by the biofilm thickness and was a function of substrate concentration only. [source] Effect of Tetrandrine on Calcium-Dependent Tumour Necrosis Factor-, Production in Glia-Neurone Mixed CulturesBASIC AND CLINICAL PHARMACOLOGY & TOXICOLOGY, Issue 4 2005Bin Wang Tetrandrine has protective effect on neuronal cells, however, the mechanisms involved in its action have not been clearly established. The aim of this study was to investigate the role of tetrandrine on calcium-dependent tumour necrosis factor-, production in glia-neurone mixed cultures. Glia-neurone mixed cultures were treated by addition of Ca2+ regulating agents for a period of 6 hr. Tetrandrine or/and TMB-8 were added 30 min. before the stimulation. The supernatant tumour necrosis factor-, levels were quantified by enzyme-linked immunosorbent assay. Exposure of lipopolysaccharide 10 and 100 ng/ml caused significant increase in tumour necrosis factor-, production respectively, with no alteration in cultures treated with 1 ng/ml lipopolysaccharide. Glia-neurone mixed cultures exhibited a marked elevation in tumour necrosis factor-, production after exposure to CaCl2, KCl, thapsigargin, BHQ and norepinephrine in the presence of lipopolysaccharide at 1 ng/ml respectively. Tetrandrine 0.3, 1, and 3 ,M concentration-dependently reduced tumour necrosis factor-, production evoked by CaCl2 or KCl. Tetrandrine preincubation had no significant effect on the response to Ca2+ -ATPase inhibitor thapsigargin or BHQ. Norepinephrine-induced tumour necrosis factor-, production was significantly reduced by tetrandrine and almost abolished by combination of tetrandrine and intracellular Ca2+ release inhibitor TMB-8. These results suggested that tetrandrine at a concentration of 0.3, 1, or 3 ,M inhibited tumour necrosis factor-, production induced by Ca2+ entry in glia-neurone mixed cultures. [source] Neurone-to-astrocyte communication by endogenous ATP in mixed culture of rat hippocampal neurones and astrocytesDRUG DEVELOPMENT RESEARCH, Issue 1 2003Schuichi Koizumi ATP is recognized as an important intercellular signaling molecule in the peripheral and CNS. Glutamate is reported to be an important neurone-to-glia mediator being released from neurones and astrocytes that activates astrocytic and neuronal Ca2+ responses, respectively. We demonstrate here that endogenous ATP could be an extracellular molecule for neurone-to-astrocyte communication in cocultured rat hippocampal neurones and astrocytes. Hippocampal neurones reveal synchronized Ca2+ oscillation, which was due to glutamatergic synaptic transmission. When analyzed in a fura-2 method, a slight and very slow increase in intracellular Ca2+ concentration ([Ca2+]i) elevation was observed in some population of astrocytes. Such astrocytic [Ca2+]i elevation was dramatically inhibited by apyrase, though apyrase itself had no effect on neuronal Ca2+ oscillation. For a detail analysis, we investigated changes in [Ca2+]i in cells using a confocal microscopy. When cocultured hippocampal neurones and astrocytes were depolarized electronically in the presence of glutamate-receptor antagonists, a transient elevation in [Ca2+]i was observed in neurones, which was followed by a slowly initiated and small rise in [Ca2+]i in astrocytes. Apyrase or P2 receptor antagonists almost abolished the [Ca2+]i rises in astrocytes, suggesting that depolarization-evoked ATP release from neurones should produce astrocytic [Ca2+]i elevation via P2 receptors. Using a luciferin,luciferase bioluminescence assay, we found that neurones could release ATP in an activity-dependent manner. These findings suggest that endogenous ATP should be an important intercellular mediator between neurones and astrocytes and that functions of these cells should be fine-tuned by endogenously released ATP in situ. Drug Dev. Res. 59:88,94, 2003. © 2003 Wiley-Liss, Inc. [source] Long-term stability of biological denitrification process for high strength nitrate removal from wastewater of uranium industryENVIRONMENTAL PROGRESS & SUSTAINABLE ENERGY, Issue 3 2008Prashant M. Biradar Abstract The aim of the present study was to biologically denitrify uranium nitrate raffinate (UNR) from nuclear industry, which is a principle source of high strength nitrate waste. To denitrify the high nitrate waste, a pilot-scale continuous stirred tank reactor was designed with two inbuilt settlers. Acclimatization of mixed culture with synthetic waste was carried out prior to the inoculation of the acclimatized sludge into the reactor. Initial concentration of nitrate in uranium raffinate was 77,000 mg/L NO3. It was diluted and used as a feed to the reactor. Concentration of nitrate in feed was increased gradually from 10,000 mg/L NO3 to 40,000 mg/L NO3 with hydraulic retention time (HRT) maintained at 34.4 h. Complete denitrification of 40,000 mg/L NO3 was achieved in a specified HRT. To facilitate understanding of the treatablity and long-term stability of biological denitrification of UNR, study was carried out for 211 days by periodical perturbation of the system. Furthermore, to find the volume ratio of reactor to settler required for the full-scale design of the denitrification plant, settling of acclimatized sludge was carried out. © 2008 American Institute of Chemical Engineers Environ Prog, 2008 [source] Impact of five selected xenobiotics on isolated ammonium oxidizers and on nitrifying activated sludgeENVIRONMENTAL TOXICOLOGY, Issue 4 2006S. N. Dokianakis Abstract Sewage treatment plants (STPs) are usual receptors of xenobiotic compounds that have to be cotreated with municipal wastewaters before being discharged to the water environment. The presence of organic contaminants, such as surfactants, polycyclic aromatic hydrocarbons (PAHs), phthalates, and their primary degradation products in the influents of STPs may inhibit irreversibly sensitive biological processes, such as nitrification. The first step of nitrification, i.e., the oxidation of ammonium to nitrite (nitritification), is particularly sensitive. Inhibition of this step under uncontrolled conditions may completely inhibit biological nitrogen removal. The aim of this work was to study the possible inhibitory effect of five selected xenobiotics on (a) a mixed culture of ammonium-oxidizing bacteria isolated from activated sludge and (b) nitrifying activated sludge directly. The xenobiotics that were tested include nonylphenols (NP), nonylphenolethoxylates (NPEO), linear alkylbenzene sulfonates (LAS), di(2-ethylhexyl) phthalate (DEHP), as a representative phthalate ester, and the PAH phenanthrene. Remarkable inhibitory effects for all tested compounds were observed in this study even at xenobiotic concentrations as low as 1 mg/L. The observed inhibition of xenobiotics on nitrifying activated sludge was less pronounced, because of the masking effect exerted by the sludge flocs, but was still significant for many of the tested substances at concentrations up to 10 mg/L. © 2006 Wiley Periodicals, Inc. Environ Toxicol 21: 310,316, 2006. [source] Nutrient dependent effects of consumer identity and diversity on freshwater ecosystem functionFRESHWATER BIOLOGY, Issue 1 2008ANDREW R. DZIALOWSKI Summary 1. Over the past decade, ecologists have tried to determine how changes in species composition and diversity affect ecosystem structure and function. Until recently, the majority of these studies have been conducted in terrestrial ecosystems and have not taken into account environmental variability. The purpose of this research was to determine how species identity and diversity in the freshwater zooplankton affected biomass of algae and zooplankton at two levels of nutrient enrichment. 2. Several species of cladocerans were grown alone and together in microcosms at both ambient and raised phosphorus concentrations to determine if the effects of consumer identity and diversity were nutrient dependent. 3. Total zooplankton biomass was greater, while algal biomass was lower, in mixed culture than in monoculture. The effects of zooplankton diversity on algal biomass, however, were only observed at raised phosphorus concentrations, suggesting that diversity effects were nutrient dependent. Specifically, diversity effects appeared to be related with biological mechanisms such as complementarity in resource use and/or facilitation. 4. More diverse communities of zooplankton appear to be better able to control algae than single species of zooplankton at high nutrient concentrations; therefore, zooplankton diversity may provide a buffer against eutrophication in freshwater ecosystems. [source] Astroglia-mediated effects of uric acid to protect spinal cord neurons from glutamate toxicityGLIA, Issue 5 2007Yangzhou Du Abstract Uric acid (UA) has been demonstrated to reduce damage to neurons elicited by oxidative stress. However, our studies utilizing cultures derived from embryonic rat spinal cord indicate that an astroglia-mediated mechanism is involved in the effects of UA to protect neurons from glutamate toxicity. The damage elicted by glutamate to neurons in a mixed culture of spinal cord cells can be reversed by UA. Furthermore, addition of UA after the termination of glutamate exposure suggests that UA plays an active role in mediating neuroprotection rather than purely binding peroxynitrite, as previously thought. Importantly, in pure neuron cultures from the same tissue, UA does not protect against glutamate toxicity. Addition of astroglia to the pure neuron cultures restores the ability of UA to protect the neurons from glutamate-induced toxicity. Our results also suggest that glia provide EAAT-1 and EAAT-2 glutamate transporters to protect neurons from glutamate, that functional EAATs may be necessary to mediate the effects of UA, and that treatment with UA results in upregulation of EAAT-1 protein. Taken together, our data strongly suggest that astroglia in mixed cultures are essential for mediating the effects of UA, revealing a novel mechanism by which UA, a naturally produced substance in the body, may act to protect neurons from damage during insults such as spinal cord injury. © 2007 Wiley-Liss, Inc. [source] Effect of different liquid cultures of live yeast strains on performance, ruminal fermentation and microbial protein synthesis in lambsJOURNAL OF ANIMAL PHYSIOLOGY AND NUTRITION, Issue 6 2008M. K. Tripathi Summary Three yeast strains, Kluyveromyces marximanus NRRL-3234 (KM), Saccharomyces cerevisiae NCDC-42 (SC) and Saccharomyces uvarum ATCC-9080 (SU), and a mixed culture (1:1:1 ratio) were evaluated for their value as probiotics in lamb feeding in two experiment. In experiment I and II, 20 and 30 pre-weaner lambs were fed for 63 and 60 days in two and three equal groups respectively. All lambs were offered ad libitum a creep mixture and Zizyphus nummularia leaves, and yeasts were dosed orally. In experiment I, one group received no yeast, the other of the mixed culture (1.5,2 × 1010 live cells/ml). In experiment II, yeast cultivation was modified yielding 1.5,2 × 1013 live cells/ml. Lambs of the three experimental groups received 1 ml/kg live weight of one of the individual yeasts. Feed intake did not differ among groups of both experiments with the exception of SC-supplemented lambs in experiment II which showed a trend to higher intakes per kg metabolic body weight and in percentage of body weight when compared with KM- and SU-supplemented lambs. Supplementation of the mixed yeast culture had no effect on intakes of digestible crude protein and metabolisable energy, nutrient digestibility, nitrogen balance and rumen fermentation characteristics (pH, ammonia, volatile fatty acid concentration, protozoa count) and urinary allantoin as an indicator of microbial protein synthesis. The same was true for comparisons in experiment II except ciliate protozoa counts, which showed a trend to be the highest with SU and the lowest with SC. The results of present study show that the response of lambs to supplemented live yeast cultures is inconsistent, as it lacked to have an effect in the present study, and that differences among strains were small, even when supplemented at a much higher live cell count. [source] Effects of thymol and diphenyliodonium chloride against Campylobacter spp. during pure and mixed culture in vitroJOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2009R.C. Anderson Abstract Aims:, To determine if the purported deaminase inhibitors diphenyliodonium chloride (DIC) and thymol reduce the growth and survivability of Campylobacter. Methods and Results:, Growth rates of Campylobacter jejuni and Camp. coli were reduced compared to unsupplemented controls during culture in Muellar,Hinton broth supplemented with 0·25 ,mol DIC or thymol ml,1 but not with 0·01 ,mol monensin ml,1 or 1% ethanol. Recovery of Camp. jejuni and Camp. coli was reduced >5 log10 CFU from controls after 24 h pure culture in Bolton broth supplemented with 0·25 or 1·0 ,mol DIC ml,1 or with 1·0 ,mol thymol ml,1. Similarly, each test Campylobacter strain was reduced >3 log10 CFU from controls after 24 h mixed culture with porcine faecal microbes in Bolton broth supplemented with 0·25 or 1·0 ,mol DIC ml,1 or with 1·0 ,mol thymol ml,1. Treatments with 0·25 ,mol thymol ml,1, 0·01 ,mol monensin ml,1 or 1% ethanol were less effective. Ammonia production during culture or incubation of cell lysates was reduced by 0·25 or 1·0 ,mol DIC ml,1 but only intermittently reduced, if at all, by the other treatments. Conclusions:, Diphenyliodonium chloride and thymol reduced growth, survivability and ammonia production of Camp. jejuni and Camp. coli. Significance and Impact of the Study:, Results suggest a potential physiological characteristic that may be exploited to develop interventions. [source] Premature Salmonella Typhimurium growth inhibition in competition with other Gram-negative organisms is redox potential regulated via RpoS inductionJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2004E. Komitopoulou Abstract Aims:, To identify the role of oxidation,reduction (redox) potential in the premature growth inhibition and RpoS induction in Salmonella serotype Typhimurium in competitive growth experiments. Methods and Results:, Oxidation,reduction potential was measured throughout the growth of a minority population of Salm. Typhimurium in mixed cultures with other Gram-negative and Gram-positive organisms. A lux -based reporter was also used to evaluate RpoS activity in Salm. Typhimurium in competitor studies. In a mixed culture, the multiplication of a minority population of Salm. Typhimurium was inhibited when competing Gram-negative organisms entered the stationary phase. This was not seen when the competing flora was Gram-positive. The change in redox potential during growth in mixed cultures was closely linked to the inhibition of Salm. Typhimurium growth by Gram-negative competitors. An artificially induced drop in redox potential earlier during growth in mixed cultures with Gram-negative organisms reduced the time to RpoS induction in Salm. Typhimurium and thus inhibited its multiplication prematurely. In contrast, RpoS induction and growth inhibition were prevented under high redox potential conditions. Conclusions:, This work shows that the inhibitory activity of competitive organisms can be mediated through their effect on redox potential-regulated RpoS induction. Significance and Impact of the Study:, Redox potential is shown to be an important determinant of Salm. Typhimurium growth, an observation with practical implications both for its control and detection. [source] VIRULENCE RESPONSE OF A SALMONELLA TYPHIMURIUM HILA:LACZY FUSION STRAIN TO SPENT MEDIA FROM PURE CULTURES OF SELECTED BACTERIA AND POULTRY CECAL MIXED CULTUREJOURNAL OF FOOD SAFETY, Issue 3 2002J.D. NUTT ABSTRACT Virulence gene expression in Salmonella is triggered by a variety of environmental factors including changes in the gastrointestinal environment of birds during different dietary regimes. The objective of this study was to determine if growth of specific microorganisms alters the environmental conditions sufficiently to signal Salmonella Typhimurium virulence response. Spent media was obtained from a Salmonella Typhimurium hilA:lacZY fusion strain, a poultry Salmonella Typhimurium strain, Eschcrichia coli K12, and Lactobacillus caseii Spent media samples were collected after 2, 4, 8 and 24 h of growth in brain heart infusion broth (BHI) and M9 media, ,-galactosidase assays were performed on the samples to determine virulence expression. Virulence response to Salmonella, spent media was 2-fold greater than Lactobacillus spent media at 4, 8 and 24 h growth (P < 0.05). Virulence expression almost doubled when exposed to Salmonella Typhimurium (NONA) spent media compared to mixed culture spent media at 4 h, and Salmonella Typhimurium (NONA) was significantly higher than mixed culture spent media at 24 h (P < 0.05). Based on these results, it appears that growth of similar bacterial species may alter the composition of rich media sufficiently to influence virulence. [source] DIEL RHYTHM OF ALGAL PHOSPHATE UPTAKE RATES IN P-LIMITED CYCLOSTATS AND SIMULATION OF ITS EFFECT ON GROWTH AND COMPETITION1JOURNAL OF PHYCOLOGY, Issue 4 2002Chi-Yong Ahn Oscillations in the phosphate (Pi) uptake rates for three species of green algae were examined in a P-limited cyclostat. For Ankistrodesmus convolutus Corda and Chlorella vulgaris Beyerinck, the Pi uptake rates increased during the daytime and decreased at night. In contrast, Chlamydomonas sp. exhibited the opposite uptake pattern. Cell densities also oscillated under a light:dark cycle, dividing at a species-specific timing rather than continuously. In general, the cell densities exhibited an inverse relationship with the Pi uptake rates. A competition experiment between A. convolutus and C. vulgaris in a P-limited cyclostat resulted in the dominance of C. vulgaris, regardless of the relative initial cell concentrations. Chlorella vulgaris also dominated in a mixed culture with Chlamydomonas sp., irrespective of the initial seeding ratio and dilution rate. However, Chlamydomonas sp. and A. convolutus coexisted in the competition experiment with gradual decrease of Chlamydomonas sp. when equally inoculated. Mathematical expressions of the oscillations in the Pi uptake rate and species-specific cell division gate were used to develop a simulation model based on the Droop equation. The simulation results for each of the species conformed reasonably well to the experimental data. The results of the competition experiments also matched the competition simulation predictions quite well, although the experimental competition was generally more delayed than the simulations. In conclusion, the model simulation that incorporated the effect of diel rhythms in nutrient uptake clearly demonstrated that species diversity could be enhanced by different oscillation patterns in resource uptake, even under the condition of limitation by the same resource. [source] Ethanol Effects on Nitric Oxide Production in Cerebral Pial CulturesALCOHOLISM, Issue 4 2001Chin-Lung Shih Background: Although alcohol abusers are known to have higher incidences of hemorrhagic cerebrovascular diseases, it is not known whether these changes are associated with ethanol (EtOH) action on nitric oxide (NO) production in the cerebrovascular cells. The purpose of this study was to examine the effects of EtOH treatment on basal and cytokine-induced NO production in cortical pial cultures. Methods: Cell cultures for this study included murine primary pial vascular cells, primary glial cells and cortical neurons. These cells were exposed to cytokines or EtOH for 24 to 48 hr. The culture media were used for measurement of nitrite, as an indication for NO release, and lactate dehydrogenase (LDH), as an index of cell membrane integrity. In addition, immunocytochemical determinations were carried out to identify cell types and to assess inducible nitric oxide synthase (iNOS). Results: Exposure of primary pial vascular cultures to cytokines that consisted of interleukin-1, (IL-1,; 250 pg/mL) and interferon-, (IFN,; 2 ng/mL) or to EtOH (50 to 100 mM) for 24 to 48 hr significantly elevated NO production. NO production could be attenuated by N -nitro-L-arginine (N-arg), a nonspecific NOS inhibitor, or aminoguanidine (AG), an iNOS inhibitor. Increased iNOS immunoreactivity was observed in cytokines- or EtOH-treated pial cells. When pial cells were cocultured with cortical neurons, prolonged EtOH exposure led to a large increase in NO production as well as LDH release. However, this increase was not observed in pial culture alone or in mixed cortical culture. Nevertheless, inhibition of NO production with N-arg or AG did not alter the EtOH-induced LDH release in the pial cells cocultured with cortical neurons. Conclusion: These results show that EtOH exposure led to increased production of NO in primary pial cell culture. In mixed culture that contained cortical neurons and pial cells, EtOH induced increase in NO as well as LDH release, which is an indication of loss of cell membrane integrity. However, EtOH-mediated LDH release in mixed cortical pial cultures was not a consequence of the increase in NO production by these cells. Studies that use mixed cortical-pial cultures may provide a unique in vitro system for examining the interactions among glial cells, neurons, and cerebrovascular cells. [source] A differential medium for lactic acid-producing bacteria in a mixed cultureLETTERS IN APPLIED MICROBIOLOGY, Issue 6 2008H.M. Lee Abstract Aims:, Modified deMan-Rogosa Sharpe agar containing bromophenol blue (mMRS-BPB) was tested as a medium for counting and differentiation of each lactic acid-producing bacterium (LAB), especially in a mixed culture. Methods and Results:, Type strains of 10 LAB species (Lactobacillus acidophilus, L. brevis, L. bulgaricus, L. gasseri, L. paracasei, L. plantarum, L. reuteri, Weissella confusa, Bifidobacterium bifidum and B. infantis) and five commercial yogurts were inoculated on plate count agar with bromocresol purple, mMRS, and mMRS-BPB. Each type strain showed more clearly formed colonies on the three media under anaerobic conditions than under aerobic conditions. Especially each type strain produced colonies with specific characteristics of each species on mMRS-BPB. Commercial yogurts produced the largest number of colonies with various shapes and colours on mMRS-BPB. Conclusions:, Modified deMan-Rogosa Sharpe agar containing bromophenol blue under anaerobic conditions is appropriate for counting and differentiating each LAB in a mixed culture. Significance and Impact of the Study:, Modified deMan-Rogosa Sharpe agar containing bromophenol blue will be useful in isolation and enumeration of each LAB from fermented foods as well as intestinal microflora. [source] Bioconversion of 3,-acetoxypregna-5,16-diene-20-one to androsta-1,4-diene-3,17-dione by mixed bacterial cultureLETTERS IN APPLIED MICROBIOLOGY, Issue 2 2002S. Patil Aims:,To isolate a bacterium capable of degrading 3,-acetoxypregna-5,16-diene-20-one (16-DPA) to androsta-1,4-diene-3,17-dione (ADD) and to decipher the biodegradation pathway. Methods and Results:,Isolation on mineral salt agar containing 16-DPA as sole carbon source yielded two bacteria identified as Pseudomonas diminuta and Comamonas acidovorons. These bacteria failed to degrade 16-DPA individually in pure cultures but converted 16-DPA to ADD in a mixed culture. The intermediates accumulated during the bioconversion were identified as pregna-4,16-diene-3,20-dione and pregna-1,4,16-triene-3,20-dione. Conclusions:,The degradation pattern of 16-DPA by mixed bacterial culture revealed the reaction sequence as (i) cleavage of C-3 acetyl function, (ii) dehydrogenation at C-1 and C-2 positions and (iii) cleavage of C-17 side-chain. Significance and Impact of the Study:,The present work opens a new approach towards the production of a female sex hormone precursor and elucidates the biodegradation pathway of 16-DPA by mixed bacterial culture. [source] Bacteriological Findings and Hormonal Profiles in the Postpartum Balady GoatsREPRODUCTION IN DOMESTIC ANIMALS, Issue 1 2006M M Ababneh Contents Twenty-six Balady goats categorized according to parity into primiparous and pluriparous goats were used to investigate bacterial flora of the genital tract and hormonal profiles during the postpartum (PP) period. Escherichia coli and Staphylococcus aureus were isolated in pure or mixed culture from the uterus. Arcanobacterium pyogenes was isolated from swabs obtained from the vagina and cervix of one primiparous goat. Uteri and cervices but not vaginas were free of bacterial contamination by day 10 PP except for one pluriparous goat with scanty E. coli contamination on day 25 PP. Fluctuating oestradiol 17, (E2) levels demonstrated resumption of follicular activity as early as day 13 PP in both parity groups. Progesterone (P4) levels remained low at basal levels throughout the study period. Higher concentrations of 15-keto-13,14-dihydroprostaglandin F2, (PGFM) were observed during the first week PP compared with the rest of the PP period. PGFM concentrations dropped to low basal level by day 10 PP and remained constantly low throughout the study period. P4, E2 and PGFM profiles were not different between the different parity groups. In conclusion, intrauterine infection is not common in goats with normal kidding. E. coli was the most common intrauterine bacterial isolate. E2 and P4 profiles were consistent with resumption of follicular growth but not ovulation. High PGFM concentrations coincided with the fast regression phase of uterine involution. Hormonal profile and bacterial contamination and clearance were similar to those reported in other related species and not related to parity. [source] Life-cycle kinetic model for endospore-forming bacteria, including germination and sporulationBIOTECHNOLOGY & BIOENGINEERING, Issue 5 2009Seongjun Park Abstract We develop a mechanistic life-cycle model for endospore-forming bacteria (EFB) and test the model with experiments with a Bacillus mixed culture. The model integrates and quantifies how sporulation and germination are triggered by depletion or presence of a limiting substrate, while both substrates affect the rate of vegetative growth by a multiplicative model. Kinetic experiments show the accumulation of small spherical spores after the triggering substrate is depleted, substantially more rapid decay during sporulation than for normal decay of vegetative cells, and a higher specific substrate utilization rate for the germinating cells than that for growth of vegetative cells. Model simulations capture all of these experimental trends. According to model predictions, when a batch reactor is started, seeding with EFB spores instead of active EFB delays the onset of rapid chemical oxygen demand (COD) utilization and biomass growth, but the end points are the same. Simulated results with low aeration intensity show that germination can consume some substrate without dissolved oxygen (DO) depletion. Biotechnol. Bioeng. 2009; 104: 1012,1024. © 2009 Wiley Periodicals, Inc. [source] The utilization of glycogen accumulating organisms for mixed culture production of polyhydroxyalkanoatesBIOTECHNOLOGY & BIOENGINEERING, Issue 4 2009*Article first published online: 15 JUN 200, Simon Bengtsson Abstract Production of polyhydroxyalkanoates (PHAs) by an open mixed culture enriched in glycogen accumulating organisms (GAOs) under alternating anaerobic,aerobic conditions with acetate as carbon source was investigated. The culture exhibited a stable enrichment performance over the 450-day operating period with regards to phenotypic behavior and microbial community structure. Candidatus Competibacter phosphatis dominated the culture at between 54% and 70% of the bacterial biomass throughout the study, as determined by fluorescence in situ hybridization. In batch experiments under anaerobic conditions, PHA containing 3-hydroxybutyrate (3HB) and 27,mol-% 3-hydroxyvalerate (3HV) was accumulated up to 49% of cell dry weight utilizing the glycogen pool stored in the SBR cycle. Under aerobic and ammonia limited conditions, PHA comprising only 3HB was accumulated to 60% of cell dry weight. Glycogen was consumed during aerobic PHA accumulation as well as under anaerobic conditions, but with different stoichiometry. Under aerobic conditions 0.31 C-mol glycogen was consumed per consumed C-mol acetate compared to 0.99 under anaerobic conditions. Both the PHA biomass content and the specific PHA production rate obtained were similar to what is typically obtained using the more commonly applied aerobic dynamic feeding strategy. Biotechnol. Bioeng. 2009; 104: 698,708 © 2009 Wiley Periodicals, Inc. [source] A Kinetic Model for Suspended and Attached Growth of a Defined Mixed CultureBIOTECHNOLOGY PROGRESS, Issue 3 2005Kawai Tam Kinetic experiments were carried out in a semicontinuous wastewater treatment process called self-cycling fermentation (SCF) using a defined mixed culture and various concentrations of synthetic brewery wastewater. The same consortium, which had been previously identified as Acinetobacter sp., Enterobacter sp., and Candida sp., were used in these experiments. The overall rate of substrate removal was attributable to both suspended microbes and the biofilm that formed during the treatment process. A rate expression was developed for the SCF system for a range of synthetic wastewaters containing glucose and various initial concentrations of ethanol and maltose. The data indicated that substrate removal by the suspended cells was directly related to the biomass concentration. However, substrate removal by the biofilm was apparently not affected by the biofilm thickness and was a function of substrate concentration only. [source] Mixed Culture Bioconversion of 16-Dehydropregnenolone Acetate to Androsta-1,4-diene-3,17-dione: Optimization of ParametersBIOTECHNOLOGY PROGRESS, Issue 2 2003Tushar Banerjee Bioconversion of 16-dehydropregnenolone acetate (16-DPA) to androsta-1,4-diene-3,17-dione (ADD), an intermediate for the production of female sex hormones, by mixed culture of Pseudomonasdiminuta MTCC 3361 and Comamonas acidovorans MTCC 3362 is reported. Various physicochemical parameters for the bioconversion of 16-DPA to ADD have been optimized in shake flask cultures. Nutrient broth inoculated with actively growing co-culture proved ideal for bacterial growth and bioconversion. A temperature range of 35,40 °C was most suitable; higher or lower temperatures adversely affected the bioconversion. Dimethylformamide below 2% concentration was the most suitable carrier solvent. Maximum conversion was recorded at 0.5 mg mL,1 16-DPA. A pH of 5.0 yielded a peak conversion of 62 mol % in 120 h incubation period. Addition of 9,-hydroxylase inhibitors failed to prevent further breakdown of ADD to nonsteroidal products. 16-DPA conversion in a 5 L fermenter followed a similar trend. [source] Effect of Inoculum Composition and Low KCl Supplementation on the Biological and Rheological Stability of an Immobilized-Cell System for Mixed Mesophilic Lactic Starter ProductionBIOTECHNOLOGY PROGRESS, Issue 6 2001L. Lamboley Two strains of Lactococcus lactissubsp. lactis (L. lactis KB and KBP) and one of L. lactissubsp. lactis biovar. diacetylactis (L. diacetylactis MD) were immobilized separately in ,-carrageenan-locust bean gum gel beads. Continuous fermentations were carried out in supplemented whey permeate in a 1-L pH-controlled stirred tank reactor inoculated with a 30% (v/v) bead inoculum and a bead ratio of 55:30:15 for KB, KBP, and MD, respectively. The process demonstrated a high productivity and microbial stability during the 7-week continuous culture. Compared with previous experiments carried out with an inoculum bead ratio of 33:33:33 for KB, KBP, and MD beads, respectively, the modification of the inoculum bead ratio had apparently little effect on free and immobilized, total and specific populations. A dominant behavior of L. diacetylactis MD over the other strains of the mixed culture was observed both with free-cell populations in the effluent and with immobilized-cell populations. Additional experiments were carried out with other strain combinations for continuous inoculation-prefermentation of milk. The data also confirmed the dominance of L. diacetylactis during long-term continuous immobilized-cell fermentations. This dominance may be tentatively explained by the local competition involved in the development of the bead cross-contamination and in citrate utilization by L. diacetylactis strains. The gel beads demonstrated a high rheological stability during the 7-week continuous fermentation even at low KCl supplementation of the broth medium (25 mM KCl). [source] Ammonia Removal Using Hepatoma Cells in Mammalian Cell CulturesBIOTECHNOLOGY PROGRESS, Issue 5 2000Yeon Sook Choi It was examined whether hepatocyte cell lines can be used for ammonia removal in mammalian cell cultures. It was found that there exists a critical ammonium concentration level for each hepatocyte cell to remove ammonia. Among the cells tested in this work, primary hepatocytes showed the strongest ammonia removal capability if ammonium concentration is higher than the critical level. However, primary hepatocytes lost the liver function gradually and finally died after 2,3 weeks. Because of this limitation, primary hepatocytes were not appropriate to be used for ammonia removal in long-term cultures. Hep G2 cells, which are immortal, also showed a strong ammonia removal activity. The ammonia removal activity of Hep G2 cells depended on the concentration of ammonium in the medium, as in the case of primary hepatocytes. However, urea could not be detected in the course of ammonia removal by Hep G2 cells. Instead of urea, Hep G2 cells secreted glutamine into the culture medium. The capacity for ammonia removal was higher in the absence than in the presence of glutamine. Thus we checked the activity of glutamine synthetase in the Hep G2 cells. The level of glutamine synthetase activity increased with the addition of ammonium chloride. This result accounts for the ammonium concentration dependency of Hep G2 cells in ammonia removal and glutamine synthesis. Furthermore Hep G2 cells could grow well in the absence of glutamine, which was necessarily required in mammalian cell cultures. These results prove that glutamine formation serves as the primary mechanism of detoxifying ammonia in hepatocyte cell lines as expected. In addition, it was demonstrated that ammonium level could be reduced 38% and that erythropoietin production increased 2-fold in the mixed culture of Hep G2 and recombinant CHO cells. [source] Kinetik des anaeroben Glycerinabbaus mithilfe sulfatreduzierender Bakterien,.CHEMIE-INGENIEUR-TECHNIK (CIT), Issue 10 2010Kinetics of Anaerobic Biodegradation of Glycerol by Sulfate-Reducing Bacteria Anaerobic glycerol degradation; Anaerobic wastewater treatment; biological sulfate reduction; heavy metals and sulphate removal; industrial sewage; Sulphate-reducing bacteria Abstract Die Wirtschaftlichkeit der Produktion von Biosulfid für die Schwermetall- oder Sulfatelimination aus Industrieabwässern hängt ausschlaggebend vom verwendeten Substrat ab und steht in direktem Zusammenhang mit der Verwertbarkeit der Kohlenstoffquelle durch Mikroorganismen. Leicht zugänglich und einfach in der Handhabung ist Glycerin als Substrat für die sulfatreduzierenden Bakterien (SRB). Gegenstand der hier vorgestellten Arbeiten war die Untersuchung des anaeroben Glycerinabbaus in der Batch-Fermentation mithilfe einer Mischkultur von SRB und die Ermittlung eines kinetischen Modellansatzes. The efficiency of the production of Biosulfid for the elimination of heavy metals or sulphate from industrial wastewater is crucial on the used substrate and is directly related to the recoverability of the carbon source by microorganisms. Easily accessible and easy to use is glycerol as substrate for the sulfate-reducing bacteria (SRB). Subject of the presented work was the investigation of the anaerobic degradation of glycerol in the batch fermentation. A mixed culture of SRB and a kinetic model approach was determined. [source] Thermodynamic Analysis of Energy Transfer in Acidogenic CulturesENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 5 2008J.-R. Bastidas-Oyanedel Abstract A global thermodynamic analysis, normally used for pure cultures, has been performed for steady-state data sets from acidogenic mixed cultures. This analysis is a combination of two different thermodynamic approaches, based on tabulated standard Gibbs energy of formation, global stoichiometry and medium compositions. It takes into account the energy transfer efficiency, ,, together with the Gibbs free energy dissipation, ,Go, analysis of the different data. The objective is to describe these systems thermodynamically without any heat measurement. The results show that , is influenced by environmental conditions, where increasing hydraulic retention time increases its value all cases. The pH effect on , is related to metabolic shifts and osmoregulation. Within the environmental conditions analyzed, , ranges from 0.23 for a hydraulic retention time of 20,h and pH,4, to 0.42 for a hydraulic retention time of 8,h and a pH ranging from 7,8.5. The estimated values of ,Go are comparable to standard Gibbs energy of dissipation reported in the literature. For the data sets analyzed, ,Go ranges from ,1210,kJ/molx, corresponding to a stirring velocity of 300,rpm, pH,6 and a hydraulic retention time of 6,h, to ,20744,kJ/molx for pH,4 and a hydraulic retention time of 20,h. For average conclusions, the combined approach based on standard Gibbs energy of formation and global stoichiometry, used in this thermodynamic analysis, allows for the estimation of Gibbs energy dissipation values from the extracellular medium compositions in acidogenic mixed cultures. Such estimated values are comparable to the standard Gibbs energy dissipation values reported in the literature. It is demonstrated that , is affected by the environmental conditions, i.e., stirring velocity, hydraulic retention time and pH. However, a relationship that relates this parameter to environmental conditions was not found and will be the focus of further research. [source] The Composition of Jerusalem Artichoke (Helianthus tuberosus L.) Spirits Obtained from Fermentation with Bacteria and YeastsENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 1 2005K. Szambelan Abstract The composition of spirits distilled from fermentation of Jerusalem artichoke (Helianthus tuberosus L.) tubers was compared by means of gas chromatography. The microorganisms used in the fermentation processes were the bacterium Zymomonas mobilis, strains,3881 and 3883, the distillery yeast Saccharomyces cerevisiae, strains,Bc16a and D2 and the Kluyveromyces fragilis yeast with an active inulinase. The fermentation of mashed tubers was conducted using a single culture of the distillery yeast Saccharomyces cerevisiae and the bacterium Zymomonas mobilis (after acid or enzymatic hydrolysis) as well as Kluyveromyces fragilis (sterilized mashed tubers). The tubers were simultaneously fermented by mixed cultures of the bacterium or the distillery yeast with K.,fragilis. The highest ethanol yield was achieved when Z.,mobilis,3881 with a yeast demonstrating inulinase activity was applied. The yield reached 94,% of the theoretical value. It was found that the distillates resulting from the fermentation of mixed cultures were characterized by a relatively lower amount of by-products compared to the distillates resulting from the single species process. Ester production of 0.30,2.93,g/L, responsible for the aromatic quality of the spirits, was noticed when K.,fragilis was applied for ethanol fermentation both in a single culture process and also in the mixed fermentation with the bacterium. Yeast applied in this study caused the formation of higher alcohols to concentrations of 7.04,g/L much greater than those obtained with the bacterium. The concentrations of compounds other than ethanol obtained from Jerusalem artichoke mashed tubers, which were fermented by Z.,mobilis, were lower than those achieved for yeasts. [source] Physiological and molecular characterization of anaerobic benzene-degrading mixed culturesENVIRONMENTAL MICROBIOLOGY, Issue 2 2003Ania C. Ulrich Summary Nine distinct anaerobic benzene-degrading cultures were enriched from sediment samples from four different sites. These cultures used nitrate, sulphate or CO2 as electron acceptors. The shortest doubling times were observed in nitrate-reducing cultures, although cell yield was lowest in these cultures. The highest substrate concentration utilized and maximum absolute rates of benzene degraded (in µM day,1) were observed in methanogenic cultures. The microbial compositions of a methanogenic and nitrate-reducing culture were determined from a clone library of 16S rRNA genes. Five Bacterial 16S rRNA sequences, one of which resembled a clone previously found in a sulphate-reducing, benzene-degrading culture and four Archaeal 16S rRNA sequences were identified in a methanogenic culture. Four Bacterial and no Archaeal 16S rRNA sequences were identified in a nitrate-reducing culture. The relative abundance of the four nitrate-reducing putative species was determined by slot blot hybridization. Two green sulphur bacteria together formed 52% of the clone library, but were found to be less than 4% of the culture by slot blot analysis. One of the cloned 16S rRNA gene sequences comprised 70% of the culture and was phylogenetically 93% similar to both Azoarcus and Dechloromonas species, which have been shown to degrade aromatic compounds, including benzene, under nitrate-reducing conditions. [source] Acute toxicity of heavy metals to acetate-utilizing mixed cultures of sulfate-reducing bacteria: EC100 and EC50ENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 12 2001Vivek P. Utgikar Abstract Acid mine drainage from abandoned mines and acid mine pit lakes is an important environmental concern and usually contains appreciable concentrations of heavy metals. Because sulfate-reducing bacteria (SRB) are involved in the treatment of acid mine drainage, knowledge of acute metal toxicity levels for SRB is essential for the proper functioning of the treatment system for acid mine drainage. Quantification of heavy metal toxicity to mixed cultures of SRB is complicated by the confounding effects of metal hydroxide and sulfide precipitation, biosorption, and complexation with the constituents of the reaction matrix. The objective of this paper was to demonstrate that measurements of dissolved metal concentrations could be used to determine the toxicity parameters for mixed cultures of sulfate-reducing bacteria. The effective concentration, 100% (EC100), the lowest initial dissolved metal concentrations at which no sulfate reduction is observed, and the effective concentration, 50% (EC50), the initial dissolved metal concentrations resulting in a 50% decrease in sulfate reduction, for copper and zinc were determined in the present study by means of nondestructive, rapid physical and chemical analytical techniques. The reaction medium used in the experiments was designed specifically (in terms of pH and chemical composition) to provide the nutrients necessary for the sulfidogenic activity of the SRB and to preclude chemical precipitation of the metals under investigation. The toxicity-mitigating effects of biosorption of dissolved metals were also quantified. Anaerobic Hungate tubes were set up (at least in triplicate) and monitored for sulfate-reduction activity. The onset of SRB activity was detected by the blackening of the reaction mixture because of formation of insoluble ferrous sulfide. The EC100 values were found to be 12 mg/L for copper and 20 mg/L for zinc. The dissolved metal concentration measurements were effective as the indicators of the effect of the heavy metals at concentrations below EC100. The 7-d EC50 values obtained from the difference between the dissolved metal concentrations for the control tubes (tubes not containing copper or zinc) and tubes containing metals were found to be 10.5 mg/L for copper and 16.5 mg/L for zinc. Measurements of the turbidity and pH, bacterial population estimations by means of a most-probable number technique, and metal recovery in the sulfide precipitate were found to have only a limited applicability in these determinations. [source] Synthetic bacterial lipopeptide analogs facilitate naive CD4+ T,cell differentiation and enhance antigen-specific HLA-II-restricted responsesEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 8 2005Mascia Ghielmetti Abstract Synthetic di- and tri-palmitoylated bacterial lipopeptide analogs (BLpA) can enhance HLA-I-restricted immune responses. Here we show that BLpA indirectly promote antigen-driven differentiation of naive CD4+ T,lymphocytes in vitro, with mechanisms that require DC and are inhibited by CTLA-4/Ig. In mixed cultures of cord blood-derived PBMC and allogeneic DC, P3CSK4 lipopeptide facilitated the transition from CCR7+/CD45RA+/CD62L+ to CCR7,/CD45RA,/CD62Ldim T,cells with kinetics significantly exceeding those obtained with the unlipidated CSK4 analog. Moreover, P3CSK4 and P2CSK4, but neither the mono-palmitoylated PCSK4 analog nor the CSK4 peptide, increased the frequency of IFN-,-producing T,cells expanded under similar conditions. Along with this, P2CSK4 and P3CSK4, but not PCSK4, restored the in vitro antigenicity of MDP-OspA, a non-immunogenic analog of Borrelia burgdorferi major outer surface lipoprotein,A, and enhanced the frequency of in vitro expanded T,cells specific for the tetanus toxoid (TT) and hepatitis,B surface antigen (HBsAg) peptides TT947,967 and HBsAg19,33 and for TT. Altogether, BLpA bearing at least two ester-bonded palmitoyl side chains indirectly enhance antigen-driven CD4+ T,cell differentiation. BLpA adjuvanticity is independent of covalent bonding to Ag and Ag formulation. This information may be helpful to generate more potent recombinant vaccines. [source] Metabolic profiling as a tool for revealing Saccharomyces interactions during wine fermentationFEMS YEAST RESEARCH, Issue 1 2006Kate S. Howell Abstract The multi-yeast strain composition of wine fermentations has been well established. However, the effect of multiple strains of Saccharomyces spp. on wine flavour is unknown. Here, we demonstrate that multiple strains of Saccharomyces grown together in grape juice can affect the profile of aroma compounds that accumulate during fermentation. A metabolic footprint of each yeast in monoculture, mixed cultures or blended wines was derived by gas chromatography , mass spectrometry measurement of volatiles accumulated during fermentation. The resultant ion spectrograms were transformed and compared by principal-component analysis. The principal-component analysis showed that the profiles of compounds present in wines made by mixed-culture fermentation were different from those where yeasts were grown in monoculture fermentation, and these differences could not be produced by blending wines. Blending of monoculture wines to mimic the population composition of mixed-culture wines showed that yeast metabolic interactions could account for these differences. Additionally, the yeast strain contribution of volatiles to a mixed fermentation cannot be predicted by the population of that yeast. This study provides a novel way to measure the population status of wine fermentations by metabolic footprinting. [source] | |||||||||||||||||||||||||||||||||||||