J Agric Food Chem, 2003 Jan 15, 51(2), 424 - 32 Chemical and sensorial aroma characterization of freshly distilled Calvados . 1 . Evaluation of quality and defects on the basis of key odorants by olfactometry and sensory analysis; Guichard H et al.; Eight freshly distilled samples of Calvados, a fermented and distilled apple juice, were analyzed by sensory evaluation and direct injection GC to determine the composition of higher alcohols, esters, and aldehydes . The composition determined by direct injection was tentatively related to sensory descriptors . Esters have a probable maximum level around 500 g/hl of pure alcohol (PA) . This level also corresponds to the threshold of the main ester constituent, ethyl acetate . A high ratio of esters to ethyl acetate seems to be of prime importance for good quality . Total aldehydes, with a maximum level between 8 and 11 g/hl of PA and mainly comprising acetal (maximum between 5 and 9 g/hl of PA), were related to a "green" descriptor . Higher alcohols do not have a direct impact on quality, but other volatile compounds with a positive impact on flavor should probably be present at a high level . As overall quality was not well related to sensory quality, it was necessary to perform more a precise analysis to determine the key odorants . The Calvados samples were thus extracted using pentane . Gas chromatography, employing both a flame ionization detector and an olfactometry port, was used to analyze the obtained extracts . Seventy-one odors were detected and distributed according to Calvados quality determined by sensory evaluation . Nineteen odors common to all Calvados samples constituted the "skeleton" of the aroma . Twenty-eight odors were specific to a quality class: 6 for good quality, 4 for neutral, and 18 for defective . Twenty-four other odors had either too low an odor impact or no evident specificity. Biosci Rep, 2002 Jun-Aug, 22(3-4), 421 - 30 F0 cysteine, bCys21, in the Escherichia coli ATP synthase is involved in regulation of potassium uptake and molecular hydrogen production in anaerobic conditions; Mnatsakanyan N et al.; The single cysteine in the b subunit of the membranous F0 sector and the 19 cysteines in extramembranous F1 sector of the Escherichia coli ATP synthase were replaced by alanine . When cells were grown under anaerobic conditions on glucose, the kcat for ATP hydrolysis of membrane vesicles containing the bCys21Ala mutant enzyme, but not enzymes with other cysteine replacements, was lower, while ATP-driven H+ pumping was unchanged . However, the ATP-dependent increase in the number of accessible thiol groups in membrane vesicles was negated . Furthermore, K+ uptake and molecular hydrogen production by whole cells and protoplasts was greatly decreased . These results indicate a role for the F0 subunit bCys21 in the functionality of F0F1 and coupling to other membranous activities under fermentative conditions. J Am Diet Assoc, 1998 Feb, 98(2), 182 - 6, 189; quiz 187-8 Nutrition and oral health guidelines for pregnant women, infants, and children; Fitzsimons D et al.; Good oral health care and nutrition during pregnancy, infancy, and childhood are essential but often overlooked factors in the growth and development of the teeth and oral cavity . Pregnant women and parents and caregivers of infants and children often receive little guidance about proper preventive dental and oral health care, including fluoride and dietary measures . Pregnant women can maintain their health through proper diet, good oral hygiene, and appropriate use of fluoride . An adequate diet during gestation is important for optimal oral development of the fetus . To promote good oral health in infancy, caregivers need to provide the infant with appropriate amounts of fluoride in addition to a healthful diet . As the teeth erupt into the mouth, the caregiver needs to clean the teeth thoroughly on a daily basis . When solid foods are introduced in later infancy, it is also important to limit the frequency of caries-promoting fermentable carbohydrates between meals . Good oral hygiene habits and dietary practices that emphasize minimum exposure to retentive, fermentable carbohydrates; use of fluoridated water; and a varied, balanced diet should continue throughout childhood to set the stage for optimal oral health for a lifetime. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi, 1998 Jun, 12(2), 139 - 42 {The study on purification and characterization of HIV-1 reverse transcriptase from a recombinant strain of E . coli}; Jia H et al.; Of the HIV proteins, reverse transcriptase(RT) has been probably the most useful target protein for screening and designing of its specific inhibitors . Because retroviral replication is absolutely dependent on both the RNase H and the polymerase function of RT and, so far as is now known, RT does not play a direct role in the life cycle of a normal cell . Under suitable fermentation conditions in our experiments, HIV-1 RT was highly expressed in E . coli JM109(pKRT-2)* by inducing the trc promoter with isopropyl-beta-Dthiogalactopyranoside(IPTG) . 1 . 1 mg of purified RT was obtained from one liter culture of bacteria by DEAE-cellulose and phosphaellulose chromatography . SDS-PAGE analysis of the purified RT showed two major protein bands of 66 kD and 51 kD, indicating that the purified RT was a heterodimer composed of two subunits . Results of enzyme assay showed that the purified RT had high activity(1.4 x 10(4) umit/mg) . We also improved the reaction system of enzyme assay . The effect of PFA on HIV-1 RT was determined with the improved enzyme assay and the mechanism of inhibition was non-competitive with respect to substrate consistent with the reports of Dr . Bo Oberg . This suggests that the purified HIV-1 RT by this simple method can be applied to the anti HIV-1-drug screening . (*E . coli JM109(pKRT2) was obtained from NIAID, NIH; pKRT2 from Dr . Richard D'Aquila and Dr . William C . Summers.) Biotechnol Bioeng, 2003 Mar 5, 81(5), 525 - 34 Effect of deletion of chitin synthase genes on mycelial morphology and culture viscosity in Aspergillus oryzae; Muller C et al.; The objective of this study was to quantify the effect of disrupting two chitin synthases, chsB and csmA, on the morphology and rheology during batch cultivation of Aspergillus oryzae . The rheological properties were characterized in batch cultivations at different biomass concentrations (from 3.4-22.5 g kg(-1) biomass) and the power-law model adequately described the rheological properties . In the cultivations there were pellets, clumps, and freely dispersed hyphal elements . The different morphological fractions were quantified using image analysis . The apparent viscosity of the fermentation broth was significantly affected by the biomass concentration, the morphology, and also by pH . The chsB disruption strain had lower consistency index K values for all biomass concentrations investigated, which is a desirable trait for industrial Aspergillus fermentations . J Nutr, 2003 Jan, 133(1), 1 - 4 Effects of dietary fibers on magnesium absorption in animals and humans; Coudray C et al.; There is overwhelming evidence that dietary fibers are an important component of human and animal diets and play an important role in human health . Because dietary fibers and some associated substances, such as phytate, have in vitro mineral-binding capacities, they have been thought to impair absorption of minerals such as calcium, iron and zinc, although magnesium absorption seems to be less affected . Indeed, the effect of dietary fibers depends largely on their own nature and characteristics, and also on mineral homeostasis . In 1977 it was observed that resistant starch, a fermentable dietary fiber, could improve Mg absorption in rats . More recently, attention has been focused on other fermentable substrates such as inulin and oligo- or polysaccharides, for their potential prebiotic and health effects . Studies conducted on different types of fermentable carbohydrates have confirmed their beneficial effect on Mg absorption in different animal species . The majority of these studies have also sought to determine the effects of fibers on other minerals such as calcium, iron and zinc . In contrast to the studies with Mg, these studies did not show a consistent effect on the absorption of these minerals . This is due to the particularities of sites and mechanism of Mg absorption . To date, four human studies have been carried out that generally confirmed the enhancing effect of fermentable oligo- or polysaccharides on Mg absorption. Appl Environ Microbiol, 2003 Jan, 69(1), 227 - 32 Purification and characterization of a prolyl aminopeptidase from Debaryomyces hansenii; Bolumar T et al.; A prolyl aminopeptidase (PAP) (EC 3.4.11.5) was isolated from the cell extract of Debaryomyces hansenii CECT12487 . The enzyme was purified by selective fractionation with protamine and ammonium sulfate, followed by two chromatography steps, which included gel filtration and anion-exchange chromatography . The PAP was purified 248-fold, with a recovery yield of 1.4% . The enzyme was active in a broad pH range (from 5 to 9.5), with pH and temperature optima at 7.5 and 45 degrees C . The molecular mass was estimated to be around 370 kDa . The presence of inhibitors of serine and aspartic proteases, bestatin, puromycin, reducing agents, chelating agents, and different cations did not have any effect on the enzyme activity . Only iodoacetate, p-chloromercuribenzoic acid, and Hg(2+), which are inhibitors of cysteine proteases, markedly reduced the enzyme activity . The K(m) for proline-7-amido-4-methylcoumarin was 40 micro M . The enzyme exclusively hydrolyzed N-terminal-proline-containing substrates . This is the first report on the identification and purification of this type of aminopeptidase in yeast, which may contribute to the scarce knowledge about D . hansenii proteases and their possible roles in meat fermentation. Appl Environ Microbiol, 2003 Jan, 69(1), 113 - 21 Oxygen consumption by anaerobic Saccharomyces cerevisiae under enological conditions: effect on fermentation kinetics; Rosenfeld E et al.; The anaerobic growth of the yeast Saccharomyces cerevisiae normally requires the addition of molecular oxygen, which is used to synthesize sterols and unsaturated fatty acids (UFAs) . A single oxygen pulse can stimulate enological fermentation, but the biochemical pathways involved in this phenomenon remain to be elucidated . We showed that the addition of oxygen (0.3 to 1.5 mg/g {dry mass} of yeast) to a lipid-depleted medium mainly resulted in the synthesis of the sterols and UFAs required for cell growth . However, the addition of oxygen during the stationary phase in a medium containing excess ergosterol and oleic acid increased the specific fermentation rate, increased cell viability, and shortened the fermentation period . Neither the respiratory chain nor de novo protein synthesis was required for these medium- and long-term effects . As de novo lipid synthesis may be involved in ethanol tolerance, we studied the effect of oxygen addition on sterol and UFA auxotrophs (erg1 and ole1 mutants, respectively) . Both mutants exhibited normal anaerobic fermentation kinetics . However, only the ole1 mutant strain responded to the oxygen pulse during the stationary phase, suggesting that de novo sterol synthesis is required for the oxygen-induced increase of the specific fermentation rate . In conclusion, the sterol pathway appears to contribute significantly to the oxygen consumption capacities of cells under anaerobic conditions . Nevertheless, we demonstrated the existence of alternative oxygen consumption pathways that are neither linked to the respiratory chain nor linked to heme, sterol, or UFA synthesis . These pathways dissipate the oxygen added during the stationary phase, without affecting the fermentation kinetics. J Dairy Sci, 2002 Dec, 85(12), 3454 - 61 Effects of a monensin controlled-release capsule or premix on attenuation of subacute ruminal acidosis in dairy cows; Mutsvangwa T et al.; The effects of monensin, administered either as a controlled release capsule (CRC) or a premix, on attenuating grain-induced subacute ruminal acidosis (SARA) and on ruminal fermentation characteristics in Holstein cows receiving a total mixed ration were investigated in two experiments . In both experiments, six multiparous, rumen-fistulated Holstein cows were used in a two-treatment, two-period crossover design with 6-wk periods . In Experiment 1, treatments were either a monensin CRC or a placebo CRC . In Experiment 2, treatments were either a monensin premix or a placebo premix . In both experiments, at the beginning of wk 4 SARA was induced in experimental cows for a 10-d period with a grain challenge model, and ruminal pH was measured continuously using indwelling pH probes . The administration of monensin either as a CRC or a premix had no effect on ruminal pH characteristics . Neither monensin CRC nor premix had an effect on ruminal volatile fatty acid concentrations, but reduced the acetate:propionate ratio . Monensin premix-treated cows were observed to have increased milk yield, largely as a result of a higher dry matter intake in monensin-treated cows compared to control cows . Milk fat content and yield were lower in monensin-treated cows compared to placebo-treated cows during SARA . In conclusion, there is no evidence that monensin was efficacious in raising ruminal pH during SARA under the conditions employed in this study. J Dairy Sci, 2002 Dec, 85(12), 3411 - 9 Effects of cultivars on ensiling characteristics, chemical composition, and ruminal degradability of pea silage; Mustafa AF et al.; A study was conducted to determine the effects of cultivar on ensiling characteristics, chemical composition and ruminal nutrient degradability of pea (Pisum sativum L.) silage . The cultivars evaluated were Lenca (L), Carneval (C), and Delta (D) . Peas were field-grown and forage was harvested and ensiled in mini-silos for 0, 2, 4, 8, 16, and 70 d . The ensiled forage of all cultivars went through a rapid fermentation with a sharp reduction in pH during the first 2 days of ensiling . Extensive proteolysis took place between 0 and 2 d as indicated by a reduction in true protein and neutral detergent insoluble protein (NDICP) and an increase in nonprotein nitrogen . Chemical analysis of the 70 d silage showed that cultivar L contained higher neutral detergent fiber (NDF) and acid detergent fiber and lower starch levels than C and D . Crude protein was highest for C (20.5% DM), intermediate for D (19.0% DM) and lowest for L (17.9% DM) . Distribution of protein fractions showed that L contains lower soluble protein and higher NDICP levels than the other two pea cultivars . However, no difference in acid detergent insoluble protein levels was observed between the three cultivars . Results of the in situ incubation experiment indicated that L had lower ruminal DM (69.2 vs 74.0%) and CP (84.1 vs 90.6%) degradabilities than C or D . However, ruminal degradability of NDF was similar among the three cultivars (average of 32.9%) . It was concluded that chemical composition and ruminal nutrient degradability of pea silage are significantly influenced by cultivars. J Dairy Sci, 2002 Dec, 85(12), 3336 - 51 Effect of protein source on amino acid supply, milk production, and metabolism of plasma nutrients in dairy cows fed grass silage; Korhonen M et al.; This study conducted according to a 4 x 4 Latin square with 28 d periods and four ruminally cannulated Finnish Ayrshire cows investigated the effect of protein supplements differing in amino acid (AA) profile and rumen undegradable protein content on postruminal AA supply and milk production . Mammary metabolism of plasma AA and other nutrients were also studied . The basal diet (Control; 13.4% crude protein) consisted of grass silage and barley in a ratio of 55:45 on a dry matter basis . The other three isonitrogenous diets (17.0% crude protein) were control + fishmeal (FM), control + soybean meal (SBM), and control + corn gluten meal (CGM) . The protein supplements replaced portions of dry matter of the control diet maintaining the silage to barley ratio constant for all diets . Dry matter intake was limited to 95% of the preexperimental ad libitum intake and was similar (mean 19.8 kg/d dry matter) across the diets . Protein supplements increased milk, lactose, and protein yields but did not affect yields of energy-corrected milk or milk fat . Milk protein yield response was numerically lowest for diet SBM . Protein supplements increased milk protein concentration but decreased milk fat and lactose concentrations . Microbial protein synthesis and rumen fermentation parameters were similar across the diets, except for an increased rumen ammonia concentration for diets supplemented with protein feeds . Protein supplements increased N intake, ruminal organic matter and N, and total tract organic matter, N, and neutral detergent fiber digestibilities . Protein supplements also increased N and AA flows into the omasum, with SBM giving the lowest and CGM the highest flows . This was associated with an unchanged microbial N flow and a higher undegraded dietary N flow . The omasal flows of individual AA reflected differences in total N flow and AA profile of the experimental diets . Differences in AA flows did not always reflect plasma AA concentrations . The results indicated that AA supply of dairy cows fed a grass silage-cereal diet can be manipulated using protein supplements differing in ruminal protein degradability and AA profile . Lower milk production response to SBM than that to FM and CGM appeared to be related mainly to lower N and AA supplies arising from a high ruminal protein degradability of SBM . Histidine appeared to be the first limiting AA for milk protein synthesis on the control diet . Mammary gland may regulate AA uptake according to requirements. Ann Bot (Lond), 2003 Jan, 91 Spec No, 279 - 90 Dynamic aspects of alcoholic fermentation of rice seedlings in response to anaerobiosis and to complete submergence: relationship to submergence tolerance; Boamfa EI et al.; Rice plants are severely damaged by complete submergence . This is a problem in rice farming and could be the result, in part, of tissue anoxia imposed by a reduced availability of oxygen . To investigate this possibility we monitored alcoholic fermentation products as markers for tissue anaerobiosis using sensitive laser-based spectroscopy able to sense ethanol and acetaldehyde down to 3 nl l(-1) and 0.1 nl l(-1), respectively . Acetaldehyde emission began within 0.5 h of imposing an oxygen-free gas phase environment followed closely by ethanol . As treatment progressed, ethanol output increased and came to exceed acetaldehyde emission as this stabilized considerably after approx . 3 h . On re-entry of air, a sharp post-anaerobic peak of acetaldehyde production was observed . This was found to be diagnostic of a preceding anoxic episode of 0.5 h or more . When anaerobiosis was lengthened by up to 14 h, the size of the post-anaerobic acetaldehyde outburst increased . After de-submergence from oxygen-free water, a similarly strong but slower post-anaerobic acetaldehyde upsurge was seen, which was accompanied by an increase in ethanol emission . Light almost, but not completely, eliminated fermentation in anaerobic surroundings and also the post-anaerobic or post-submergence peaks in acetaldehyde production . All photosynthetically generated oxygen was consumed within the plant . There was no substantial difference in acetaldehyde and ethanol output between FR13A and the less submergence-tolerant line CT6241 under any submergence treatment . In some circumstances, submergence damaged CT6241 more than FR13A even in the absence of vigorous fermentation . We conclude that oxygen deprivation may not always determine the extent of damage caused to rice plants by submergence under natural conditions. Ann Bot (Lond), 2003 Jan, 91 Spec No, 195 - 204 Assessment of enzyme induction and aerenchyma formation as mechanisms for flooding tolerance in Trifolium subterraneum 'Park'; Aschi-Smiti S et al.; The objective of this study was to evaluate the role of enzyme induction and aerenchyma formation in prolonged tolerance to soil flooding in a variety of underground clover (Trifolium subterraneum 'Park') previously selected for resistance . Seedlings were grown in hydroponic tanks, initially with aeration for 3 weeks and subsequently in the absence of aeration for up to 3 weeks . After 1 h in the absence of aeration, the oxygen concentration in the hydroponic medium had decreased to 1.5 % . During the 3 weeks of extreme oxygen deficiency, primary roots died and were replaced by considerable numbers of adventitious roots . Activities of many glycolytic and fermentative enzymes increased in adventitious roots . Excised adventitious roots were capable of immediate induction of ethanol in the absence of lactate production, in association with energy charge higher than that in excised roots of aerobically maintained controls . Energy charge was even higher when measured in adventitious roots in planta . Interestingly, haemoglobin protein could be correlated with energy charge . Aerenchyma was readily visualized in adventitious roots by optical microscopy of longitudinal and transverse sections . We conclude that avoidance of root anoxia via aerenchyma is the major mechanism for prolonged root tolerance in Trifolium subterraneum 'Park'. Ann Bot (Lond), 2003 Jan, 91 Spec No, 155 - 72 Functional electron microscopy in studies of plant response and adaptation to anaerobic stress; Vartapetian BB et al.; This article reviews the contribution made by functional electron microscopy towards identifying and understanding the reactions of plant roots and shoots to anaerobic stress . Topics examined include: (1) unexpected hypersensitivity, rather than hyper-resistance, to anoxia of root tips of flooding-tolerant plants; (2) protective, rather than damaging, effects of a stimulated energy metabolism (glycolysis and fermentation) under anaerobic conditions; (3) the concept of two main strategies of plant adaptation to anaerobic environments, namely avoidance of anaerobiosis on the whole plant level, termed 'apparent' tolerance, and metabolic adaptation at the cellular and molecular levels, termed 'true' tolerance; (4) the importance of protein synthesis during hypoxia and anoxia for enhanced energy production and metabolic adaptation; (5) a general adaptive syndrome in plants to stress at the ultrastructural level and a possible molecular mechanism for its realization under anoxia; (6) the physiological role of anaerobically synthesized lipids and nitrate as alternative electron acceptors in an oxygen-free medium; and (7) the selection of cell lines derived from callus cultures that possess enhanced tolerance to anoxia and can regenerate whole plants with improved tolerance of soil waterlogging. Int J Syst Evol Microbiol, 2002 Nov, 52(Pt 6), 2315 - 9 Citeromyces siamensis sp . nov., a novel halotolerant yeast isolated in Thailand; Nagatsuka Y et al.; Two halotolerant yeast strains, H130(T) and H149, were isolated from dry salted squid and fermented soybeans, respectively, in Thailand . Both isolates grew by multilateral budding, produced asci that had one roughened spherical ascospore and contained ubiquinone Q-8 . These characteristics were shared by Citeromyces matritensis, the only species of the genus Citeromyces . Strains H130(T) and H149 were differentiated from C matritensis by their ability to assimilate L-sorbose and L-lysine and to grow at 37 degrees C . The novel isolates were more tolerant to higher concentrations of cations (3 M NaCl or 0.8 M LiCI) and to higher osmotic pressure (60% glucose) than C . matritensis . A phylogenetic analysis of 18S rRNA gene sequence data indicated that the two novel isolates represented a sister species to C . matritensis . Furthermore, DNA-DNA hybridization data indicated that the isolates were clearly distinct from the type strain of C . matritensis (IFO 0954(T) . Based on the above characteristics, strains H130(T) and H149 are proposed to represent a novel species within the genus Citeromyces, Citeromyces siamensis; the type strain is H130(T) (= IFO 11052(T) = JCM 11522(T) = TISTR 5777(T) = CBS 9153(T)). Pediatr Res, 2003 Jan, 53(1), 113 - 8 Nicotinic induction of preproenkephalin and tyrosine hydroxylase gene expression in butyrate-differentiated rat PC12 cells: a model for adaptation to gut-derived environmental signals; Nankova BB et al.; Accelerated maturation of peripheral sympathoadrenal transmitter levels and function occurs at 7-10 postnatal days in the rat . This event is temporally disconnected from the timing of major changes in physiologic stimuli evident after the birthing process (i.e . temperature, oxygen, sound, light, etc.) . Colonization of the gut, fermentation of carbohydrates, and production of short-chain fatty acids (e.g . butyrate) mirrors this postnatal time course . In this report, we examined the interaction between butyrate differentiation of rat pheochromocytoma cells and cholinergic-nicotinic induction of the neuropeptide (enkephalin) and catecholamine-related biosynthetic enzymes (tyrosine hydroxylase, dopamine beta-hydroxylase, phenylethanolamine N-methyltransferase) . Our results show that butyrate induces both preproenkephalin and tyrosine hydroxylase mRNA through a proximal promoter region and that this regulatory step is time and dose dependent . Moreover, there is an additional interaction with cholinergic-nicotinic inducible mechanisms consistent with classically described transsynaptic cholinergic regulation of these genes . Dopamine beta-hydroxylase and phenylethanolamine N-methyltransferase promoters were not affected by butyrate treatment . We speculate that colonization of the human gut (along with the attendant fermentation of enteral carbohydrates to short-chain fatty acids) may represent a mechanism through which environmental signals affect postnatal maturation of sympathoadrenal transmitter systems. J Exp Bot, 2003 Jan, 54(382), 467 - 75 Metabolic signalling and carbon partitioning: role of Snf1-related (SnRK1) protein kinase; Halford NG et al.; A protein kinase that plays a key role in the global control of plant carbon metabolism is SnRK1 (sucrose non-fermenting-1-related protein kinase 1), so-called because of its homology and functional similarity with sucrose non-fermenting 1 (SNF1) of yeast . This article reviews studies on the characterization of SnRK1 gene families, SnRK1 regulation and function, interacting proteins, and the effects of manipulating SnRK1 activity on carbon metabolism and development. Bioresour Technol, 2003 May, 87(3), 337 - 9 Grape and apple wines volatile fermentation products and possible relation to spoilage; Polychroniadou E et al.; The main volatile by-products of the alcoholic fermentation of grape wine, cider and apple pulp wine were investigated to determine if any correlated with spoilage resistance in the latter two . Spoilage was visually detected after seven days in low-alcohol grape wine in comparison to 11 and 16 days in cider and apple pulp wine, respectively . Acetaldehyde, ethyl acetate, methanol, propanol, isobutanol and amyl alcohols were the main fermentation by-products detected in all three wines . There were highest concentrations of acetaldehyde, ethyl acetate, methanol and propanol in grape wine and, therefore, these by-products could not be implicated in spoilage resistance in apple wines . Increased concentrations of isobutanol and amyl alcohols, however, in cider and apple pulp wine in comparison to grape wine might have been the reason for spoilage resistance in the apple wines. Folia Microbiol (Praha), 2002, 47(5), 511 - 5 Influence of cultivation conditions on citrate production by Aspergillus niger in a semi-pilot-scale plant; Ul-Haq I et al.; The influence of some fermentation parameters on the semi-pilot scale (alteration of growth conditions, e.g., sugar concentration, incubation temperature and initial pH) on citrate production was demonstrated in parent and mutant strains of Aspergillus niger . Raw material from sugar industry (cane molasses) was examined as basal fermentation medium in a stirred stainless-steel 15-L fermentor . After growth on medium with 150 g/L sugar, the parent strain produced 51.2 g/L citric acid; the mutant strain achieved production maximum of 96.2 g/L . Comparing the growth, kinetic (volumetric substrate uptake rate, rate of substrate consumption and volumetric productivity rate) and production parameters it was found that the mutant strain grows more rapidly, with slightly changed morphology (intermediate, shiny round pellets with diameter 0.6-0.7 mm), and exhibits a higher citrate production and higher efficiency of sugar utilization. Folia Microbiol (Praha), 2002, 47(5), 493 - 8 Isolation and identification of Streptomyces sp . and assay of its exocellular water-soluble blue pigments; Lu L et al.; A bacterial strain producing a great amount of blue pigment during submerse fermentation was isolated and identified . Based on morphological characteristics, cell-wall chemotype and sequence of 16S rRNA gene, the strain should belong to the genus Streptomyces; it had 99.4% homology of 16S rRNA gene sequence with that of Streptomyces indigocolor . The pigment production by the strain was affected by carbon and nitrogen sources . The main components of the pigment mixture (detected by HPLC and TLC) were tentatively classified as actinorhodin-related compounds . The pigment was relatively stable against light and higher temperature but was sensitive to low pH . The preliminary acute-toxicity determination showed that the pigment was nontoxic (LD50 > 15 mg/g). Biotechniques, 2002 Dec, 33(6), 1296 - 300 Buffered non-fermenter system for lab-scale production of secreted recombinant His-tagged proteins in Saccharomyces cerevisiae; Ngamkitidechakul C et al.; Expression of recombinant proteins using a secretion system can minimize co-purification of contaminating host proteins . Production of His-tagged recombinant proteins in the yeast alpha-factor secretion system has previously required a fermenter system to control the growth conditions such as pH of the yeast culture . We describe an inexpensive non-fermenter system for the production of secreted recombinant His-tagged proteins in Saccharomyces cerevisiae that uses a buffered low peptone YP glycerol medium, which does not interfere with immobilized metal affinity chromatography . Maspin, a tumor suppressor serpin, was expressed as a secreted N-terminal His/FLAG-tagged protein . Purification of the soluble active recombinant protein only requires centrifugation, concentration by ultrafiltration, and Ni2+ affinity chromatography . Purified protein yields of this system are 3-5 mg/L culture medium. Microbiol Res, 2002, 157(4), 267 - 74 Isolation and identification of yeasts associated with vineyard and winery by RFLP analysis of ribosomal genes and mitochondrial DNA; Sabate J et al.; Yeast colonies isolated from vineyard and cellar substrates were analysed in the present study . Yeast species assessment was carried out by amplification and digestion of a region of the ribosomal RNA gene repeat unit . Saccharomyces strains were also characterised using mitochondrial DNA restriction analysis . Oxidative basidiomycetous yeasts without enological potential were predominant in the vineyard environment . Yeasts associated with grape skin depend on grape variety, vintage and degree of grape maturation . These species from grape surface constituted the predominant microbiota in must and they developed during the first stages of the process . Yeasts colonies were also isolated and identified from the walls of a fermentation vat some days before the harvest . Contray to what was expected, Saccharomyces cerevisiae was not the major species isolated as Candida sorbosa represented 76% of the species isolated . Saccharomyces strains isolated from the fermentation vat had been previously isolated in wine fermentations in this cellar . Therefore, these strains should be considered as constant residents of this winery. J Gen Appl Microbiol, 2002 Oct, 48(5), 261 - 7 Isolation and clonal pre-selection of enological Saccharomyces; Caridi A et al.; The aim of the present study was to perform a fast pre-selection from a great number of wine yeasts using a simple phenotypic-based methodology that allows many different strains to be simultaneously tested . A total of 150 elliptic yeasts, isolated from must and wine from black grapes of a distinctive Italian variety, were studied . Yeasts were identified to genus level by assessing their ability to ferment glucose and their production of spores on acetate agar . The Saccharomyces strains were seeded on BiGGY agar to determine their H(2)S production, on calcium carbonate agar to test their acetic acid production, and on grape-skin agar and on grape-seed agar to assess their interaction with phenolic compounds . The Saccharomyces strains were also examined for fermentative vigor after 2 d or 7 d both with and without the addition of 100 mg L(-1) of SO(2) in must at 20 degrees brix and pH 3.20 . At the end of fermentation, the wines produced by the 18 best yeasts were analyzed and the strains were studied for additional biochemical and technological characteristics . The resistance of the strains to simultaneous acid-stress and osmotic-stress was studied carrying out in duplicate winemaking tests in must at 30 degrees brix and pH 2.60 . A remarkable heterogeneity among the 150 autochthonous yeasts studied was demonstrated . The phenotypical biodiversity is particularly interesting for several technological characteristics useful in winemaking, such as fermentation vigor, acetic acid production and malic acid content of the wines . The vast majority of the elliptic wine yeasts isolated did not show suitable characteristics, so only 18 strains, 12% of the total, remained for the final tests . Many of the strains that had passed the preliminary screenings revealed some defects when they were studied for fermentation performance, both in standard winemaking and under stressors . Two strains exhibited particularly interesting performances: one strain for winemaking of normal musts and the other for winemaking of musts from dried grapes or under stressful conditions. J Gen Appl Microbiol, 1998 Oct, 44(5), 347 - 353 Breeding of flocculent industrial alcohol yeast strains by self-cloning of the flocculation gene FLO1 and repeated-batch fermentation by transformants; Ishida-Fujii K et al.; A nonflocculent industrial polyploid yeast strain, Saccharomyces cerevisiae 396-9-6V, was converted to a flocculent one by introducing a functional FLO1 gene at the URA3 locus . The flocculent strain FSC27 obtained was a so-called self-cloned strain, having no bacterial DNA . FSC27 cells could be easily recovered for reuse from fermentation mash without any physical energy . The strain produced a concentration of alcohol as high as 396-9-6V, although the fermentation rate of FSC27 was slightly lower than that of 396-9-6V . When uracil was added to the medium or when URA3 was reintroduced into FSC27 (named FSCU-L18), the fermentation rate and the growth rate increased, and the ethanol concentration produced was higher than that produced by the parent strain . The stable flocculation and high ethanol productivity were observed by using FSCU-L18 during 10 cycles of repeated-batch fermentation test. J Gen Appl Microbiol, 1999 Feb, 45(1), 23 - 28 Identification of acetic acid bacteria isolated from Indonesian sources, especially of isolates classified in the genus Gluconobacter; Yamada Y et al.; Sixty-four strains of acetic acid bacteria were isolated from Indonesian sources such as fruits, flowers, and fermented foods by the enrichment culture at pH 3.5 . Forty-five strains were routinely identified as Acetobacter strains because of their oxidation of acetate and lactate to carbon dioxide and water and their Q-9 isoprenolog, corresponding to 70% of all the 64 acetic acid bacteria isolated . Eight isolates were identified as Gluconacetobacter strains because of their oxidation of acetate and lactate and their Q-10 isoprenolog, occupying 13% of all the isolates . The remaining 11 isolates, accommodated in the genus Gluconobacter because of no oxidation of acetate and lactate and because of their Q-10 isoprenolog, accounted for 17% of all the isolates . They were divided into two groups based on DNA base compositions . One comprised the seven isolates, which had high G1C contents of DNA ranging from 60.3 to 63.5 mol% and of which DNAs hybridized with that of the type strain of Gluconobacter oxydans at values of 64-94% of DNA relatedness . The other comprised the remaining four isolates, which had low G+C contents of DNA ranging from 57.5 to 57.7 mol% and of which DNAs hybridized with that of the type strain of Gluconobacter frateurii at values of 63-77% of DNA relatedness . The high values of DNA relatedness, 84 to 96%, were obtained between the type strains of Gluconobacter cerinus and Gluconobacter asaii. J Gen Appl Microbiol, 1999 Feb, 45(1), 1 - 6 Isolation and characterization of triacylglycerol-secreting mutant strain from yeast, Saccharomyces cerevisiae; Nojima Y et al.; To establish the molecular bases for development of a microbiological system approaching excretive fermentation of useful lipids, a mutant strain that accumulates lipids in the medium was isolated from the laboratory yeast Saccharomyces cerevisiae . Following the mutagenesis to strain YP1, a long chain fatty acid utilizer with ethylmethane sulfonate, the mutant strain, STG1, was selected from about 80,000 colonies . The analysis of extracellular lipids and the monitoring of leakage of intracellular proteins indicated that strain STG1 secreted lipids containing triacylglycerols into the extracellular space without cell lysis . Genetic studies clarified that this mutation was recessive and was complemented by wild-type genomic DNA fragments . STG1 was considered to be a good tool for elucidation of the molecular mechanism for transmembrane lipid transport. J Gen Appl Microbiol, 1999 Jun, 45(3), 99 - 103 Production of alkaline protease by a genetically engineered Aspergillus oryzae U1521; Samarntarn W et al.; The production of alkaline protease of Aspergillus oryzae U1521 was examined in liquid culture . In a culture of defatted soybean only, it gave satisfactory enzyme yields at 584,000 U/g defatted soybean . When various carbohydrates were supplemented, enzyme production was significantly increased . An increase in production by lactose was the most marked . Enrichment with casitone or casein increased productivity, but not cornsteep solid . Media formulation (g/L) of defatted soybean 10, lactose 5, casitone 1, and KH(2)PO(4) 5 enhanced alkaline protease production by A . oryzae U1521 to a maximum of 1,410,000 U/g defatted soybean . Scaling-up experiments indicated the flask-scale results could be reproduced at 40 g of substrate in 5-L fermenter . The enzyme activity was maximum between pH 8-9 and at a temperature of 45 degrees C. J Gen Appl Microbiol, 1999 Dec, 45(6), 277 - 282 Effect of three strains of Pleurotus tuber-regium (Fr.) Sing . on chemical composition and rumen fermentation of wheat straw; Jalc D et al.; This study was conducted to investigate changes in in vitro dry matter digestibility (IVDMD) and cell wall constituent degradation in wheat straw treated with 3 strains of the fungus Pleurotus tuber-regium (PT) . The incubation of wheat straw for 30 days at 28 degrees C improved IVDMD from 30.3% (UWS-untreated wheat straw) to 47.1% for strain PT1, to 48.5% for PT4, and was unchanged IVDMD-29.9% -for PT5 . The growth of fungi was accompanied by the dry matter loss of wheat straw: 31.5% for PT1, 20.9% for PT4, and 4.8% for PT5 . Fungal treatment was characterized by increased crude protein and ash contents (%) in all fungi-treated straws and reduced hemicellulose and lignin content . It is evident that enzymes of all 3 PT strains preferentially degraded hemicellulose and lignin over cellulose . Wheat straw treated with PT1 (TWS-PT1), PT4 (TWS-PT4), and PT5 (TWS-PT5) and barley (80% : 20%) were used as the experimental diets at the fermentation in the artificial rumen . UWS with barley (80% : 20%) served as the control diet . The fermentation of experimental diets was accompanied with increased IVDMD and a very low degree of hemicellulose degradation . Total gas and methane productions were similar in all diets . Moreover, total volatile fatty acid (VFA) production (mmol day(-1)), mol % of acetate, propionate, butyrate, isobutyrate, and isovalerate were not influenced during the fermentation of experimental diets . From the stoichiometric relations, production, utilization, and recovery of metabolic hydrogen and organic matter fermented were unchanged . Only the recovery of metabolic hydrogen in TWS-PT5 was significantly increased in comparison to control diet . Total microbial production showed the tendency of lower values in experimental diets, and it was accompanied with a significant decrease of ammonia nitrogen (mg L(-1)) . Finally the results showed that the strains of Pleurotus tuber-regium can improve the quality of wheat straw, but the loss of dry matter (DM) (mainly hemicellulose) limits the effective utilization of fungi-treated straw in ruminant digestion. J Gen Appl Microbiol, 1997 Jun, 43(3), 133 - 137 Effect of six species of white-rot basidiomycetes on the chemical composition and rumen degradability of wheat straw; Jalc D et al.; This study was conducted to investigate changes in in vitro dry matter digestibility (IVDMD), volatile fatty acids (VFA) production and cell-wall constituent degradation in wheat straw treated with six white-rot fungi: Daedalea quercina, Hericium clathroides, Phelinus laevigatus, Inonotus andersonii, Inonotus obliquus, and Inonotus dryophilus . The incubation of wheat straw for 30 days at 28 C improved IVDMD from 41.4 (control) to 59.2% for D . quercina, 56.3% for H . clathroides, 50.2% for P . laevigatus, 51.4% for I . andersonii, 52% for I . obliquus, and 55.9% for I . dryophilus . In contrast, the growth of fungi was accompanied by the dry matter loss of wheat straw: 43% for D . quercina, 12% for H . clathroides, and 22-25% for the other fungi . It is evident that the increase in digestibility by D . quercina was not offset by a loss of dry matter . The total VFA production during the rumen fermentation of fungus-treated straw was slightly increased by H . clathroides and I . dryophilus only . Neutral detergent fiber (NDF) and acid detergent fiber (ADF) were reduced in fungus-treated straw . Out of the three fractions (hemicellulose, cellulose, and lignin), hemicellulose and lignin showed the largest proportionate loss after inoculation with the fungi D . quercina, H . clathroides, P . laevigatus, and I . obliquus . The other two fungi showed the largest proportionate loss in cellulose and hemicellulose contents . The results of this study suggest that the digestion enhancement of wheat straw colonized by white-rot fungi is regulated by complex factors including the degradation of structural carbohydrates and lignin. Biofizika, 2002 Nov-Dec, 47(6), 1064 - 7 {Interaction of membrane proton conductivity, membrane and oxidation-reduction potential in Escherichia coli}; Akopian K et al.; It was shown that the proton conductivity of Escherichia coli membranes depends on pH and other conditions of bacterial growth . It is considerably lower in cells fermenting glucose and accomplishing the nitrate-nitrite respiration compared with cells accomplishing the oxygen respiration . Proton conductivity increases substantially with decreasing pH of medium . It was found that proton conductivity is related to the redox and membrane potentials of cells . The energy-dependent flux of protons from cells and the ATPase activity of membrane vesicles considerably vary depending on whether bacteria are grown under aerobic or anaerobic conditions . The H+ flux from cells fermenting glucose (pH 7.5) was 1.7 times greater than the H+ flux from cells that accomplish the nitrate-nitrite and oxygen respiration . The N,N'-dicyclohexylcarbodiimide (DCCD)-sensitive ATPase activity increased 2.5 times as K+ concentration increased to 100 mM (including residual K+ in potassium-free medium) . The DCCD-sensitive ATPase activity considerably decreased with decreasing pH of medium, whereas the ATPase activity that was not suppressed by DCCD was stimulated . These results can be used for establishing the relationship between membrane proton conductivity and the energy-dependent H+ flux and ATPase activity. Br J Nutr, 2002 Dec, 88 Suppl 3, S281 - 5 Bioavailability of minerals in legumes; Sandberg AS; The mineral content of legumes is generally high, but the bioavailability is poor due to the presence of phytate, which is a main inhibitor of Fe and Zn absorption . Some legumes also contain considerable amounts of Fe-binding polyphenols inhibiting Fe absorption . Furthermore, soya protein per se has an inhibiting effect on Fe absorption . Efficient removal of phytate, and probably also polyphenols, can be obtained by enzymatic degradation during food processing, either by increasing the activity of the naturally occurring plant phytases and polyphenol degrading enzymes, or by addition of enzyme preparations . Biological food processing techniques that increase the activity of the native enzymes are soaking, germination, hydrothermal treatment and fermentation . Food processing can be optimized towards highest phytate degradation provided that the optimal conditions for phytase activity in the plant is known . In contrast to cereals, some legumes have highest phytate degradation at neutral or alkaline pH . Addition of microbial enzyme preparations seems to be the most efficient for complete degradation during processing . Fe and Zn absorption have been shown to be low from legume-based diets . It has also been demonstrated that nutritional Fe deficiency reaches its greatest prevalence in populations subsisting on cereal- and legume-based diets . However, in a balanced diet containing animal protein a high intake of legumes is not considered a risk in terms of mineral supply . Furthermore, once phytate, and in certain legumes polyphenols, is degraded, legumes would become good sources of Fe and Zn as the content of these minerals is high. Appl Biochem Biotechnol, 2003 Jan, 104(1), 23 - 36 Effect of freeze-dried immobilized cells on delignified cellulosic material in low-temperature and ambient-temperature wine making; Ikonomopoulou M et al.; In this article, we report on wine making by freeze-dried immobilized cells on delignified cellulosic material for ambient and low temperatures . Biocatalyst supported by freeze-dried delignified cellulosic (FDC) material recovered after the first repeated-batch fermentations the fermentation efficiency and startup, which become about equal to those of biocatalyst supported by wet delignified cellulosic material . The FDC biocatalyst was suitable for wine making at low temperatures (5-15 degrees C), and produced wine of 12% alcoholic degree, with the main volatiles contained in the wine and reduced by a decrease in temperature . The fermentation efficiency was not affected by total acidity of must, while an increase in initial Be density improved percentages of higher alcohols and ethyl acetate . The quality of the wine was validated by a preliminary taste test to be in the range of acceptable to excellent. J Food Prot, 2002 Dec, 65(12), 1854 - 60 Fate of Escherichia coli O157:H7 during silage fermentation; Byrne CM et al.; The survival characteristics of Escherichia coli O157:H7 in silage derived from contaminated grass were investigated . The survival of other enteric bacteria was also investigated to determine if E . coli O157:H7 demonstrates enhanced acid tolerance in comparison . Samples of chopped grass were treated as follows: (i) no additive (control); (ii) inoculation with E . coli O157:H7 to a final concentration of log10 4.0 CFU g(-1); (iii) addition of an 85% solution of formic acid at 3.0 ml kg(-1) grass; and (iv) addition of both E . coli O157:H7 and formic acid, at the above concentrations . Treated 6-kg grass samples were packed into laboratory silos, sealed, and stored at 15 degrees C for up to 180 days . Individual replicate silos were removed from storage periodically and subjected to microbiological and chemical analyses . Chemical analyses of the silage samples indicated that lactic acid-dominant fermentations, with a rapid drop in pH, occurred . Numbers of enteric bacteria decreased from log10 7.0 to 8.0 CFU g(-1) to undetectable levels within 19 days' storage . E . coli O157:H7 did not survive the silage fermentation process, with numbers declining from approximately log10 4.0 CFU g(-1) to undetectable levels within 19 days of ensiling . The pattern of decline in numbers of E . coli O157:H7 was the same as that for the enteric bacteria, indicating that under the conditions tested, the acid tolerance of E . coli O157:H7 was not significantly different from the acid tolerance of other enteric bacteria . This study found that E . coli O157:H7 did not survive a good silage fermentation process, indicating that properly ensiled grass that is correctly stored is unlikely to be a vector for the transmission of the pathogen among cattle. J Appl Microbiol, 2003, 94(1), 120 - 6 Optimization of submerged culture process for the production of mycelial biomass and exo-polysaccharides by Cordyceps militaris C738; Kim SW et al.; AIMS: The objective of the present study was to determine the optimal culture conditions for mycelial biomass and exo-polysaccharide (EPS) by Cordyceps militaris C738 in submerged culture . METHODS AND RESULTS: The optimal temperatures for mycelial biomass and EPS production were 20 degrees C and 25 degrees C, respectively, and corresponding optimal initial pHs were found to be 9 and 6, respectively . The suggested medium composition for EPS production was as follows: 6% (w/v) sucrose, 1% (w/v) polypeptone, and 0.05% (w/v) K2HPO4 . The influence of pH on the fermentation broth rheology, morphology and EPS production of C . militaris C738 was carried out in a 5-l stirred-tank fermenter . The morphological properties were comparatively characterized by pellet roughness and compactness by use of image analyser between the culture conditions with and without pH control . The roughness and compactness of the pellets indicated higher values at pH-stat culture (pH 6.0), suggesting that larger and more compact pellets were desirable for polysaccharide production (0.91 g g(-1) cell d(-1) . CONCLUSIONS: Under the optimized culture conditions (with pH control at 6), the maximum concentration of biomass and EPS were 12.7 g l(-1) and 7.3 g l(-1), respectively, in a 5-l stirred-tank fermenter . SIGNIFICANCE AND IMPACT OF THE STUDY: The critical effect of pH on fungal morphology and rheology presented in this study can be widely applied to other mushroom fermentation processes. Biotechnol Bioeng, 2003 Feb 20, 81(4), 448 - 58 Determination of in vivo oxygen uptake and carbon dioxide evolution rates from off-gas measurements under highly dynamic conditions; Wu L et al.; In vivo kinetics of Saccharomyces cerevisiae are studied, in a time window of 150 s, by analyzing the response of O(2) and CO(2) in the fermentor off-gas after perturbation of chemostat cultures by metabolite pulses . Here, a new mathematical method is presented for the estimation of the in vivo oxygen uptake rate (OUR) and carbon dioxide evolution rate (CER) directly from the off-gas data in such perturbation experiments . The mathematical construction allows effective elimination of delay and distortion in the off-gas measurement signal under highly dynamic conditions . A black box model for the fermentor off-gas system is first obtained by system identification, followed by the construction of an optimal linear filter, based on the identified off-gas model . The method is applied to glucose and ethanol pulses performed on chemostat cultures of S . cerevisiae . The estimated OUR is shown to be consistent with the independent dissolved oxygen measurement . The estimated in vivo OUR and CER provide valuable insights into the complex dynamic behavior of yeast and are essential for the establishment and validation of in vivo kinetic models of primary metabolism . Biotechnol Bioeng, 2003 Feb 20, 81(4), 438 - 47 Use of confocal microscopy to follow the development of penetrative hyphae during growth of Rhizopus oligosporus in an artificial solid-state fermentation system; Nopharatana M et al.; Two methods were compared for determining the concentration of penetrative biomass during growth of Rhizopus oligosporus on an artificial solid substrate consisting of an inert gel and starch as the sole source of carbon and energy . The first method was based on the use of a hand microtome to make sections of approximately 0.2- to 0.4-mm thickness parallel to the substrate surface and the determination of the glucosamine content in each slice . Use of glucosamine measurements to estimate biomass concentrations was shown to be problematic due to the large variations in glucosamine content with mycelial age . The second method was a novel method based on the use of confocal scanning laser microscopy to estimate the fractional volume occupied by the biomass . Although it is not simple to translate fractional volumes into dry weights of hyphae due to the lack of experimentally determined conversion factors, measurement of the fractional volumes in themselves is useful for characterizing fungal penetration into the substrate . Growth of penetrative biomass in the artificial model substrate showed two forms of growth with an indistinct mass in the region close to the substrate surface and a few hyphae penetrating perpendicularly to the surface in regions further away from the substrate surface . The biomass profiles against depth obtained from the confocal microscopy showed two linear regions on log-linear plots, which are possibly related to different oxygen availability at different depths within the substrate . Confocal microscopy has the potential to be a powerful tool in the investigation of fungal growth mechanisms in solid-state fermentation . Curr Genet, 2002 Dec, 42(3), 147 - 52 Epub 2002 Nov 29. Up-regulation of tRNA biosynthesis affects translational readthrough in maf1-delta mutant of Saccharomyces cerevisiae; Kwapisz M et al.; Maf1p is a negative effector of RNA polymerase III in yeast . The maf1-delta mutation caused an increase in the level of cellular tRNAs, but a decrease of translational readthrough at nonsense codons . Using the lacZ- luc dual gene reporter system, we detected an almost twofold diminution of UAA and UAG readthrough in maf1-delta compared with the parental strain . The maf1-delta mutation did not affect the rate of protein biosynthesis and growth at standard conditions, but resulted in temperature-sensitive growth on non-fermentable carbon sources . We examined the correlation of the temperature sensitive and antisuppression phenotypes of maf1- Delta using a colour phenotype assay in the ade2-1 SUP11 strain . Antisuppression, but not the temperature-sensitive growth defect, was compensated either by increased dosage of SUP11or by {PSI(+)}, the prion form of the translation termination factor Sup35p . Summarizing, the elevated tRNA levels in maf1- Delta increase translational fidelity and, independently, affect growth under special conditions. J Dairy Sci, 2002 Nov, 85(11), 2964 - 73 Ruminal digestion and fermentation of high-producing dairy cows with three different feeding systems combining pasture and total mixed rations; Bargo F et al.; Six multiparous Holstein cows fitted with rumen cannulas were used to study the effect of three feeding systems combining pasture and total mixed rations (TMR) on ruminal digestion in a 21-wk repeated measures experiment . The three treatments were: 1) pasture plus concentrate (PC), 2) pasture plus partial TMR (pTMR), and 3) TMR (nonpasture) . Ruminal NH3-N concentration was lower on both the pTMR and TMR treatments (10.2 +/- 0.5 mg/dL) than on the PC treatment (19.9 +/- 0.5 mg/dL) . Ruminal pH was not affected by treatments and averaged 5.87 . Neither total volatile fatty acid concentration (137.5 mmol/L) nor individual volatile fatty acid proportions (63.1,20.6, and 12.0 mol/ 100 mol for acetate, propionate, and butyrate, respectively) differed among treatments . The pTMR treatment reduced the total potentially degradable fraction of dry matter (85.5 vs . 82.3%) and the potentially digestible fraction of neutral detergent fiber (82.1 vs . 74.9%) of pasture compared to the PC treatment . Ruminal NH3-N losses were reduced when combining pasture and TMR; however this combination decreased the ruminal digestion of pasture, indicating the presence of associative effects in the rumen. J Dairy Sci, 2002 Nov, 85(11), 2928 - 47 Corn silage management III: effects of hybrid, maturity, and processing on nitrogen metabolism and ruminal fermentation; Johnson LM et al.; Two experiments were conducted to evaluate the effects of maturity and mechanical processing of two hybrids of whole plant corn silage on DM and OM digestibility, nitrogen metabolism, ruminal fermentation, and milk production and composition in lactating Holstein cows . In the first experiment, Pioneer hybrid 3845 whole plant corn was harvested at hard dough, one-third milkline, and two-thirds milkline with a theoretical length-of-cut of 6.4 mm . At each stage of maturity, corn was harvested with (1-mm roll clearance) and without (15.9-mm roll clearance) mechanical processing using a John Deere 5830 harvester with an on-board kernel processor . In the second experiment, Pioneer hybrids 3845 and Quanta were harvested at one-third milkline, two-thirds milkline, and blackline stages of maturity with and without mechanical processing . The theoretical length-of-cut was 12.7 mm . Total tract DM and OM digestibilities were lower for cows fed diets containing processed corn silage in experiment 1, and tended to be lower for cows fed diets containing unprocessed corn silage in experiment 2 . Ruminal acetate concentrations were greater and ruminal propionate concentrations were lower 2 and 6 h after feeding for cows fed diets containing corn silage harvested at physiological maturity in experiment 2 . This was due to decreased digestion of starch at advanced maturities in experiment 2 . Ruminal pH tended to decline rapidly after feeding for cows fed hybrid Quanta (2 h) compared to hybrid 3845 (5 h) corn silage based diets . Ruminal acetate concentrations decreased and ruminal propionate concentrations increased 2 and 6 h after feeding for cows fed diets containing hybrid Quanta corn silage compared to hybrid 3845 corn silage . This was related to a greater starch concentration in the corn silage, greater starch intake, and increased rate of starch digestion for cows fed hybrid Quanta corn silage-based diets . Microbial nitrogen flow was lower and feed nitrogen flow was greater for cows fed diets containing hybrid Quanta corn silage . The lower microbial nitrogen flow was due to lower microbial nitrogen concentration and nonammonia nitrogen flow to the duodenum . Milk fat and protein concentrations had a strong quadratic relationship with forage NDF intake as a percentage of body weight . When forage NDF intake as a percentage of body weight dropped below 0.70%, there was a rapid decline in milk fat and protein concentrations. J Dairy Sci, 2002 Nov, 85(11), 2890 - 904 Ruminal fermentation and nutrient digestion by dairy cows fed varying amounts of soyhulls as a replacement for corn grain; Ipharraguerre IR et al.; Five multiparous Holstein cows cannulated in the rumen and duodenum that averaged 63 d in milk were used in a 5 x 5 Latin square design with 14-d periods to evaluate the incremental substitution of soyhulls for corn in the diet . Diets contained 23% alfalfa silage, 23% corn silage, and 54% concentrate on a dry matter (DM) basis . Pelleted soyhulls replaced corn in the concentrate to supply 0, 10, 20, 30, or 40% of the dietary DM . The intakes of DM and organic matter were unaffected by treatments . Intakes of acid detergent fiber and neutral detergent fiber increased linearly, but the intake of nonstructural carbohydrates decreased linearly as soyhulls increased from 0 to 40% of dietary DM . The amount of acid detergent fiber and neutral detergent fiber digested was increased whereas the amount of nonstructural carbohydrate digested was decreased in the rumen, in the lower digestive tract, and in the total digestive tract as soyhulls replaced corn in the diet . Passage to the duodenum of nonammonia N, microbial N, nonammonia nonmicrobial N, total essential amino acids, total nonessential amino acids, and total amino acids were not affected by treatments . Yield of milk (29.5 kg/d) was not affected by treatments in this experiment . In a companion experiment, cows fed the 40% SH diet produced 1.2 kg/day per cow less (P < 0.07) milk than cows fed the control diet which is similar to the 1.3 kg/day per cow less milk produced by cows fed the same 40% SH diet in this experiment . Differences in the source of energy (fiber vs . nonstructural carbohydrates), in the amount of fiber and nonstructural carbohydrates digested, and in the site of digestion in the gastrointestinal tract may cause a shortage of the source and/or amount of energy that is required for maximum milk production in high producing cows when more than 30% of the dietary DM that is supplied as corn is replaced with soyhullss. Crit Rev Biotechnol, 2002, 22(4), 355 - 74 Production and mass transfer characteristics of non-Newtonian biopolymers for biomedical applications; Richard A et al.; The market for microbial biopolymers is currently expanding to include several emerging biomedical applications . Specifically, these applications are drug delivery and wound healing . A fundamental understanding of the key fermentation parameters is necessary in order to optimize the production of these biopolymers . Considering that most microbial biopolymer systems exhibit non-Newtonian rheology, oxygen mass transfer can be an important parameter to optimize and control . In this article, we present a critical review of recent advances in rheological and mass transfer characteristics of selected biopolymers of commercial interest in biomedical applications. Exp Biol Med (Maywood), 2002 Dec, 227(11), 1017 - 21 Modulation of CD8+ intraepithelial lymphocyte distribution by dietary fiber in the rat large intestine; Ishizuka S et al.; We studied whether ingestion of dietary fiber modifies the distribution of intraepithelial lymphocytes (IEL) in a physiological condition . Male WKAH rats were fed diets either with fiber (sugar beet fiber or crystalline cellulose, 100 g/kg diet each) or without fiber for 3 weeks . The number of CD8(+), CD4(+), and NKR-P1(+) IEL per epithelial layer in the crypt section of the cecum, proximal colon, and distal colon were scored by immunohistochemical staining . We found that the proportion of CD8(+) IEL was greater in the cecal mucosa and was gradually reduced toward the distal large intestine in general . In contrast, there was no difference in the proportion of CD4(+) and NKR-P1(+) IEL in the large intestine . Dietary sugar beet fiber, but not crystalline cellulose, increased the proportion of CD8(+) IEL, especially in the cecal mucosa, but not the CD4(+) and NKR-P1(+) IEL . Analysis of cecal organic acid concentration confirmed higher concentrations of acetate and butyrate, and lower concentration of succinate and isovalerate, in the cecum of the rats fed sugar beet fiber than other diets . These results indicate that ingestion of some dietary fiber modulates local cell proliferation of a progenitor of CD8(+) IEL or promotes homing of CD8(+) T cells into the large intestinal epithelium, most likely via the fermentation in the luminal contents. Lett Appl Microbiol, 2003, 36(1), 54 - 8 Saccharomyces cerevisiae biodiversity in spontaneous commercial fermentations of grape musts with 'adequate' and 'inadequate' assimilable-nitrogen content; Granchi L et al.; AIM: To evaluate whether intraspecific diversity of Saccharomyces cerevisiae in wine fermentations is affected by initial assimilable-nitrogen content . METHODS AND RESULTS: Saccharomyces cerevisiae isolates from two spontaneous commercial wine fermentations started with adequate and inadequate nitrogen amounts were characterized by mitochondrial DNA restriction analysis . Several strains occurred in each fermentation, two strains, but not the same ones, being predominant at frequencies of about 30% . No significant differences were detected by comparing the biodiversity indices of the two fermentations . Cluster analysis demonstrated that the strain distribution was independent of nitrogen content, the two pairs of closely related dominant strains grouping into clusters at low similarity . CONCLUSIONS: The genetic variability of S . cerevisiae in wine fermentations seemed not to depend on the nitrogen availability in must . SIGNIFICANCE AND IMPACT OF THE STUDY: Nitrogen content did not affect the genetic diversity but may have induced a 'selection effect' on S . cerevisiae strains dominating wine fermentations, with possible consequences on wine properties. J Gen Appl Microbiol, 2001 Jun, 47(3), 119 - 131 Identification of Acetobacter strains isolated from Indonesian sources, and proposals of Acetobacter syzygii sp . nov., Acetobacter cibinongensis sp . nov., and Acetobacter orientalis sp . nov . Lisdiyanti P, Kawasaki H, Seki T, Yamada Y, Uchimura T, Komagata K. Forty-six strains of acetic acid bacteria newly isolated from flowers, fruits, and fermented foods collected in Indonesia were taxonomically studied . They were Gram-negative rods, produced acetic acid from ethanol, oxidized acetate and lactate to CO(2) and H(2)O, and had Q-9 as the major ubiquinone system . On the basis of DNA-DNA similarity, all strains studied, including type strains and reference strains of the genus Acetobacter, were separated into eleven groups (Groups I to XI) . Of the 46 isolates, two isolates were included in Group II and identified as Acetobacter pasteurianus, five in Group IV as A . orleanensis, 16 in Group V as A . lovaniensis, five in Group VII as A . indonesiensis, and three in Group VIII as A . tropicalis . The remaining 15 isolates constituted three new groups based on DNA-DNA similarity; four isolates were included in Group IX, two in Group X, and nine in Group XI . No isolates were identified as A . aceti (Group I), A . peroxydans (Group III), and A . estunensis (Group VI) . Phylogenetic analysis based on 16S rDNA sequences of representative strains of the Groups indicated belonging to the strains of the genus Acetobacter . On the basis of DNA base composition, DNA-DNA similarity, and 16S rDNA sequences, three new species of the genus Acetobacter are proposed: Acetobacter syzygii sp . nov . for Group IX, Acetobacter cibinongensis sp . nov . for Group X, and Acetobacter orientalis sp . nov . for Group XI . The distribution of Acetobacter strains in Indonesia is discussed in light of isolation sources. J Gen Appl Microbiol, 2001 Jun, 47(3), 107 - 117 Structure and function of cyclin-dependent Pho85 kinase of Saccharomyces cerevisiae; Toh-E A et al.; Yeast Saccharomyces cerevisiae has five cyclin-dependent protein kinases (Cdks), Cdc28, Srb10, Kin28, Ctk1, and Pho85 . Any of these Cdks requires a cyclin partner for its kinase activity and a Cdk/cyclin complex, thus produced, phosphorylates a set of specific substrate proteins to exert its function . The cyclin partners of Srb10, Kin28, and Ctk1 are Srb11, Ccl1, and Ctk2, respectively . In contrast to the fact that each of Srb10, Kin28, and Ctk1 has a single cyclin partner, Cdc28 and Pho85 are polygamous; Cdc28 has 9 cyclins and Pho85 has 10 cyclins . Among these Cdks, Kin28 and Cdc28 are essential Cdks and it is well known that Cdc28 kinase plays a major role in regulating cell cycle progression . Pho85 is a non-essential Cdk but its absence causes a broad spectrum of phenotypes such as constitutive expression of PHO5, inability to utilize non-fermentable carbon sources, defects in cell cycle progression, and so on . Pho85 homologues are expanding to higher eukaryotes . Pho85 is most closely related with Cdk5 in terms of the amino acid sequence . The functional analysis of the domains of Pho85 also supports the close relationship between Pho85 and Cdk5, in which it was shown that the method of regulation of these two kinases is similar . Furthermore, forced expression of the mammalian CDK5 gene in a pho85Delta strain canceled a part of the pho85 defects . In this review, we summarize the functions of both Pho85/cyclin kinase and emphasize yeast Pho85 as valuable model systems to elucidate the functions of their homologues in other organisms. J Ind Microbiol Biotechnol, 2002 Dec, 29(6), 314 - 22 Influence of the yeast strain on the changes of the amino acids, peptides and proteins during sparkling wine production by the traditional method; Martinez-Rodriguez AJ et al.; The influence of five yeast strains on the nitrogen fractions, amino acids, peptides and proteins, during 12 months of aging of sparkling wines produced by the traditional or Champenoise method, was studied . High-performance liquid chromatography (HPLC) techniques were used for analysis of the amino acid and peptide fractions . Proteins plus polypeptides were determined by the colorimetric Bradford method . Four main stages were detected in the aging of wines with yeast . In the first stage, a second fermentation took place; amino acids and proteins plus polypeptides diminished, and peptides were liberated . In the second stage, there was a release of amino acids and proteins, and peptides were degraded . In the third stage, the release of proteins and peptides predominated . In the fourth stage, the amino acid concentration diminished . The yeast strain used influenced the content of free amino acids and peptides and the aging time in all the nitrogen fractions. J Hepatobiliary Pancreat Surg, 2002, 9(4), 448 - 58 Enteral nutrition in HPB surgery: past and future; Bengmark S; Perioperative nutrition has, during the past century, been transformed from a tool to provide calorie and nitrogen support to a tool to boost the immune system and increase resistance to complications . Despite all the progress in medicine and surgery, perioperative morbidity, the rate of infections, thrombosis, and the development of serosal adhesions has remained the same as long as can be judged, or at least during the past 80 years . Most prone to develop complications are persons above the age of 65 and persons with depressed immunity . About 80% of the immune system is localized in the gastrointestinal tract, which offers great opportunities for modulation through enteral nutrition . As the stomach has a tendency to develop postoperative paralysis, tube feeding is often necessary . In 1918, Andresen demonstrated the advantages of enteral nutrition, which already started on the operating table . Mulholland and colleagues and Rhoads and co-workers demonstrated, during the 1940s, certain advantages of enteral tube feeding . Also, the works by Alexander, Fischer, and Ryan, and their co-workers supported the value of early enteral feeding, and suggested enteral feeding as an effective tool to boost the immune system . It was, however, works published in the early 1990s, by Moore and colleagues and by Kudsk and colleagues, which made surgeons more aware of the advantages of early enteral nutrition . Surgery in the hepatobiliary pancreatic field is known to have a high rate of complications . Uninterrupted perioperative nutrition, i.e., nutrition during the night before, during surgery, and immediately after, offers a strong tool to prevent complications . It is essential that the nutrition also provides food for the colon, e.g., fiber and healthy bacteria (probiotics) to ferment the fiber and boost the immune system. Berl Munch Tierarztl Wochenschr, 2002 Nov-Dec, 115(11-12), 453 - 7 Effect of Tween 80 and monensin on ruminal fermentation of the diet containing 70% wheat straw treated by white-rot fungus in artificial rumen; Jalc D; The objective of this study was to investigate the effect of Tween 80 and monensin on rumen fermentation of the diet containing 70% wheat straw treated by white-rot fungus Pleurotus tuber-regium (TWS-PT) and 30% barley in artificial rumen (RUSITEC) . The RUSITEC consisted of four fermentation vessels (V1, V2, V3, V4): V1 was without additives (control), V2 received daily 10 mg of monensin, V3 received daily 0.5% Tween 80 (vol.wt-1) and V4 involved the combination of 10 mg of monensin with 0.5% Tween 80 (vol.wt-1) . After an adaptation period (6 days) the fermentation parameters were determined for six consecutive days . Tween 80 did not affect the rumen fermentation of the diet consisting 70% TWS-PT and 30% barley in RUSITEC . Monensin affected the rumen fermentation of the diet by the decreased degradability of DM, neutral detergent fiber (NDF), acid detergent fiber (ADF), hemicellulose, cellulose (p < 0.001), the decrease of methane production (p < 0.001) and the higher proportion of propionate within the volatile fatty acids (p < 0.001) in comparison to control . Tween 80 did not improve the potency of monensin . Only some indices of the increase mol% of propionate (about 3.4%) and the decrease of methane production (about 0.47 mmol.day-1) were found by using Tween 80 plus monensin in comparison to use of monensin alone. Trends Biotechnol, 2003 Jan, 21(1), 29 - 37 Accelerated design of bioconversion processes using automated microscale processing techniques; Lye GJ et al.; Microscale processing techniques are rapidly emerging as a means to increase the speed of bioprocess design and reduce material requirements . Automation of these techniques can reduce labour intensity and enable a wider range of process variables to be examined . This article examines recent research on various individual microscale unit operations including microbial fermentation, bioconversion and product recovery techniques . It also explores the potential of automated whole process sequences operated in microwell formats . The power of the whole process approach is illustrated by reference to a particular bioconversion, namely the Baeyer-Villiger oxidation of bicyclo{3.2.0}hept-2-en-6-one for the production of optically pure lactones. Surg Neurol, 2002 Sep-Oct, 58(3-4), 271 - 3 Gas-containing otogenic brain abscess; Paolini S et al.; BACKGROUND: Gas-containing brain abscesses are very rare . Two mechanisms may be responsible for the presence of intracavitary gas: bacterial fermentation or penetration through an abnormal communication between the exterior and the intracranium . The need to search for this potential communication is considered an indication for open surgery . We report the case of a surgically treated gas-containing brain abscess originating from an undiagnosed chronic otitis media . CASE DESCRIPTION: A 54-year-old man developed acute neurologic deterioration, becoming comatose within 24 hours . A contrast-enhanced computed tomography (CT) scan disclosed a gas-containing cystic mass in the right temporal lobe . Urgent surgical decompression revealed the presence of an abscess, which was excised . During the same surgery, we performed a radical mastoidectomy, removing a previously undiagnosed attic cholesteatoma . Neither procedure revealed a discontinuity of the floor of the middle cranial fossa . Cultures grew a mixed flora . Antibiotics were administered for 6 weeks . The patient made a complete neurologic recovery . CONCLUSION: This report demonstrates that otogenic brain abscesses may contain gas due to fermentation of nonclostridial bacteria. Yeast, 2002 Dec, 19(16), 1399 - 411 Characterization of a gene encoding tRNA nucleotidyltransferase from Candida glabrata; Hanic-Joyce PJ et al.; A gene encoding ATP (CTP):tRNA nucleotidyltransferase (EC2.7.7.25) was isolated from Candida (Torulopsis) glabrata by complementation in Saccharomyces cerevisiae . The predicted amino acid sequence of the protein revealed a large region with high sequence similarity to members of the Class II group of the nucleotidyltransferase superfamily and an N-terminal region characteristic of a mitochondrial targeting sequence . The essential role of the carboxylates within the conserved DXD and RRD motifs was confirmed by mutagenesis . C . glabrata strains bearing truncated CCA1 genes that lacked sequences encoding the putative mitochondrial targeting peptide were unable to grow on non-fermentable carbon sources but were able to grow on a fermentable carbon source . These results suggest that, as in S . cerevisiae, the C . glabrata CCA-adding enzyme is a sorting isozyme that functions in multiple cellular compartments . Mapping of the 5'-ends of primary transcripts of CCA1 revealed multiple transcription start sites located both upstream of and between two in-frame start codons . When the cells were cultured on a non-fermentable carbon source the longer transcripts appeared more abundant, suggesting that the choice of transcription start sites was influenced by carbon source . The shorter transcripts, which lacked sequences encoding the mitochondrial targeting information, were more predominant in cells grown on glucose . These observations suggest that expression of CCA-adding isozymes in C . glabrata may be regulated . The DNA sequence has been assigned GenBank Accession No . AF098803 . Curr Genet, 2002 Nov, 42(2), 94 - 102 Epub 2002 Nov 21. The mitochondrial genome can be altered or lost without lethal effect in the petite-negative yeast Debaryomyces (Schwanniomyces) occidentalis; Fernet CS et al.; The nature of mutations affecting several cytochrome-deficient mutants of Debaryomyces (Schwanniomyces) occidentalis has been characterized . The DR12 mutant, which is deficient in cytochrome b, and the B10Mn mutant, which is deficient in cytochromes b and a, a3, are deleted in the mitochondrial CYB and COX1 genes respectively . The B10 strain, which is partially deficient in cytochrome b, has no detectable change in its mitochondrial DNA and possibly carries nuclear lesion(s) . These three mutants, unlike the rho(-) and rho degrees "petite" mutants of Saccharomyces cerevisiae, can still grow on non-fermentable substrates, due to the development of a salicylhydroxamic acid (SHAM)-sensitive alternative pathway linked to phosphorylation at site 1 . A gly(-) mutant lacking mtDNA and respiratory capacity has been isolated . For the first time, it is demonstrated that mtDNA can be altered or even lost without lethal consequence in D . occidentalis, although this yeast was classified as a petite-negative species. Microbiol Immunol, 2002, 46(10), 667 - 75 Extracellular ATP regulates cell death of lymphocytes and monocytes induced by membrane-bound lipoproteins of Mycoplasma fermentans and Mycoplasma salivarium; Into T et al.; The cytotoxicities of lipoproteins of Mycoplasma fermentans and Mycoplasma salivarium to a lymphocytic cell line, MOLT-4, and a monocytic cell line, HL-60, was upregulated by ATP added extracellularly in a dose-dependent manner . These lipoproteins induced ATP release and plasma membrane permeability increase in these cell lines . In addition, periodate-oxidized ATP, an antagonist for P2X purinergic receptors, suppressed the cytotoxicity of the lipoproteins, suggesting the possibility that P2X receptors for ATP play crucial roles in the cytotoxicity . Activation of caspase-3 induced by the lipoproteins, which was assessed by the cleavage of the synthetic substrate DEVD-pNA and the endogenous substrate poly(ADP-ribose) polymerase, was also upregulated and downregulated by extracellular ATP and periodate-oxidized ATP, respectively . On the basis of these results, this study suggests that mycoplasmal lipoproteins induce the permeability increase in lymphocytes and monocytes, by which ATP is released, and the ATP regulates the cytotoxicities of the lipoproteins to the cells, possibly by interaction with ATP receptors such as P2X purinergic receptors. Mikrobiyol Bul, 2002 Jan, 36(1), 49 - 55 {Catalase-negative Rothia dentocariosa: evaluation of additional descriptive tests}; Ergin C et al.; Rothia dentocariosa is one of the human oral flora members, and classified in the group of coryneform bacteria which are taxonomically heterogeneous . Although R . dentocariosa is considered as a contaminant in throat cultures generally, it may be the causative infectious agent in the presence of predisposing factors . The catalase activity which is used as a simple and initial identification test, becomes contradictory because of the detection of the catalase negative strains recently . The aim of this study was to compare several biochemical and enzymatic reactions for the identification of catalase positive and negative R . dentocariosa strains . A total of 42 R . dentocariosa strains which were isolated from throat cultures have been studied, and the positivity rate of mannose fermentation in catalase negative R . dentocariosa isolates was found to be significantly higher than those catalase positive (p < 0.05) . In conclusion, mannose fermentation test seems to be a useful tool as an additional identification method of catalase negative R . dentocariosa isolates. J Agric Food Chem, 2002 Dec 18, 50(26), 7660 - 8 Volatile response of four apple varieties with different coatings during marketing at room temperature; Bai J et al.; Five experimental coatings with different resistance to gas exchange were used with freshly harvested and 20-week commercially stored apples of Delicious, Fuji, Braeburn, and Granny Smith varieties . The coated or noncoated apples were held at 20 degrees C for up to 4 weeks . The gas partial pressures inside the fruits with the various coatings ranged from 1 to 25 kPa CO(2) and from 20 to 1 kPa O(2) . Volatile evaporation rates were measured, as also were the volatiles compositions in the fruit . The coatings with intermediate gas resistance (carnauba-shellac mixture and candelilla) gave intermediate values of CO(2) and O(2) in the internal atmosphere in Delicious, Fuji, and Braeburn apples and the highest concentrations of butyl acetate and 2-methylbutyl acetate in the fruits . The coatings with the highest gas resistance (shellac and shellac-protein) caused high internal CO(2) and low O(2), resulting in anaerobic fermentation in Braeburn and Granny Smith apples and relatively high amounts of low-molecular-weight ethyl esters trapped within the fruit . A small portion of the alcohols were evaporated from fruits compared to esters, this attributed to their high Henry's law coefficients. Biotechnol Bioeng, 2003 Feb 5, 81(3), 341 - 7 Pulsed feeding during fed-batch fungal fermentation leads to reduced viscosity without detrimentally affecting protein expression; Bhargava S et al.; The goal in this study was to determine if pulsed addition of substrate could be used to alter filamentous fungal morphology during fermentation, to result in reduced broth viscosity . In all experiments, an industrially relevant strain of Aspergillus oryzae was grown in 20-liter fermentors . As a control, cultures were fed limiting substrate (glucose) continuously . Tests were performed by altering the feeding strategy so that the same total amount of glucose was fed in repeated 300-s cycles, with the feed pump on for either 30 or 150 s during each cycle . Variables indicative of cellular metabolic activity (biomass concentration, oxygen uptake rate, base consumed for pH control) showed no significant difference between continuous and pulse-fed fermentations . In addition, there was no significant difference between total extracellular protein expression or the apparent distribution of these proteins . In contrast, fungal mycelia during the second half of pulse-fed fermentations were approximately half the size (average projected area) of fungi during fermentations with continuous addition of glucose . As a result, broth viscosity during the second half of pulse-fed fermentations was approximately half that during the second half of continuous fermentations . If these results prove to be applicable for other fungal strains and processes, then this method will represent a simple and inexpensive means to reduce viscosity during filamentous fungal fermentation . Arch Microbiol, 2002 Dec, 179(1), 1 - 6 Epub 2002 Nov 07. Dissimilation of the C2 sulfonates; Cook AM et al.; Organosulfonates are widespread in the environment, both as natural products and as xenobiotics; and they generally share the property of chemical stability . A wide range of phenomena has evolved in microorganisms able to utilize the sulfur or the carbon moiety of these compounds; and recent work has centered on bacteria . This Mini-Review centers on bacterial catabolism of the carbon moiety in the C2-sulfonates and the fate of the sulfonate group . Five of the six compounds examined are subject to catabolism, but information on the molecular nature of transport and regulation is based solely on sequencing data . Two mechanisms of desulfonation have been established . First, there is the specific monooxygenation of ethanesulfonate or ethane-1,2-disulfonate . Second, the oxidative, reductive and fermentative modes of catabolism tend to yield the intermediate sulfoacetaldehyde, which is now known to be desulfonated to acetyl phosphate by a thiamin-diphosphate-dependent acetyltransferase . This enzyme is widespread and at least three subgroups can be recognized, some of them in genomic sequencing projects . These data emphasize the importance of acetyl phosphate in bacterial metabolism . A third mechanism of desulfonation is suggested: the hydrolysis of sulfoacetate. J Environ Qual, 2002 Nov-Dec, 31(6), 1795 - 801 Application technique and slurry co-fermentation effects on ammonia, nitrous oxide, and methane emissions after spreading: II . Greenhouse gas emissions; Wulf S et al.; The aim of this study was to investigate the effect of different application techniques on greenhouse gas emission from co-fermented slurry . Ammonia (NH3), nitrous oxide (N2O), and methane (CH4) emissions were measured in two field experiments with four different application techniques on arable and grassland sites . To gather information about fermentation effects, unfermented slurry was also tested, but with trail hose application only . Co-fermented slurry was applied in April at a rate of 30 m3 ha(-1) . Measurements were made every 4 h on the first day after application and were continued for 6 wk with gradually decreasing sampling frequency . Methane emissions were <150 g C ha(-1) from co-fermentation products and seemed to result from dissolved CH4 . Only in the grassland experiment were emissions from unfermented slurry significantly higher, with wetter weather conditions probably promoting CH4 production . Nitrous oxide emission was significantly increased by injection on arable and grassland sites two- and threefold, respectively . Ammonia emissions were smallest after injection or trail shoe application and are discussed in the preceding paper . We evaluated the climatic relevance of the measured gas emissions from the different application techniques based on the comparison of CO2 equivalents . It was evident that NH3 emission reduction, which can be achieved by injection, is at least compensated by increased N2O emissions . Our results indicate that on arable land, trail hose application with immediate shallow incorporation, and on grassland, trail shoe application, bear the smallest risks of high greenhouse gas emissions when fertilizing with co-fermented slurry. J Environ Qual, 2002 Nov-Dec, 31(6), 1789 - 94 Application technique and slurry co-fermentation effects on ammonia, nitrous oxide, and methane emissions after spreading: I . Ammonia volatilization; Wulf S et al.; Ammonia emissions after spreading animal manure contribute a major share to N losses from agriculture . There is an increasing interest in anaerobic co-digestion of liquid manure with organic additives . This fermentation results in a change of physical and chemical parameters of the slurry . Among these are an increased pH and ammonium content, implying a higher risk of NH3 losses from fermentation products . To compare different application techniques and the effect of fermentation on NH3 volatilization, we used the standard comparison method and tested it for reliability . This method seems to be perfectly suited for experiments with a large number of treatments and replicates if prerequisites concerning the experimental layout are considered . We tested four different application techniques on arable and grassland sites . The more the substrate was incorporated into the soil or applied near the soil surface on the grassland site, the less NH3 was lost . Injection of the substrate reduced losses to less than 10% of applied NH4+ on both sites, whereas losses after splash plate application amounted to more than 30% . Trail shoe application on grassland performed as well as injection . Harrowing on arable land also reduced emissions efficiently, if harrowing occurred within the first 2 h after application . Emissions from trail hose-applied co-fermentation product were not greater than from unfermented slurry . Better infiltration of the less viscous substrate seemed to have compensated for the increased loss potential. J Nutr, 2002 Dec, 132(12), 3599 - 602 Dietary chicory inulin increases whole-body bone mineral density in growing male rats; Roberfroid MB et al.; Chicory inulin is a natural linear fructan that is not digested in the upper part of the gastrointestinal tract but is fermented in the cecocolon . It enhances calcium absorption in rats and improves femur and tibia mineral contents in gastrectomized or ovariectomized rats . We studied the effect of inulin (0, 5 and 10 g/100 g diet) on whole-body bone mineral content (WBBMC), whole-body bone area (WBBA) and whole-body bone mineral density (WBBMD) in live, growing male rats fed diets containing 0.2, 0.5 or 1 g Ca/100 g . Three experiments, each corresponding to one of the different dietary Ca concentrations, were performed using male Wistar rats (n = 108; 4 wk old) . WBBMC was measured by dual-energy X-ray absorptiometry every 4 wk up to wk 22 . Inulin increased WBBMC (P < 0.05) and WBBMD (P < 0.001) significantly but not WBBA at all ages and all dietary calcium concentrations . This is the first report to demonstrate that chicory inulin not only increases calcium absorption but also increases mineral parameters in whole-body bones. Biotechnol Prog, 2002 Nov-Dec, 18(6), 1431 - 8 Comparison of techniques for monitoring antibody fragment production in E . coli fermentation cultures; Bowering LC et al.; The use of an optical biosensor for monitoring antibody fragment accumulation following induction in a batch fermentation of recombinant E . coli is compared to the more traditional method of ELISA quantification . Using the biosensor, concentration data can be obtained within minutes of sample addition to the device, compared to an average assay time of 3-4 h for the ELISA . We describe two biosensor assays developed as an alternative to ELISA and compare them with ELISA in the ability to provide quantitative product accumulation profiles during fermentation . Discrepancies in titers recorded by the assays are explained by a combination of differences in product variants detected by each assay and interference from sample contaminants . Method of sample preparation is also shown to be important if accurate concentration data is required . Both biosensor assays are shown to be capable of providing product accumulation profiles comparable to those obtained by ELISA . The use of a rapid extraction technique would allow such data to be obtained during process operation, enabling improved fermentation control and more rapid process development. Biotechnol Prog, 2002 Nov-Dec, 18(6), 1366 - 76 Identification of critical batch operating parameters in fed-batch recombinant E . coli fermentations using decision tree analysis; Buck KK et al.; To develop a useful fermentation process model, it is first necessary to identify which batch operating parameters are critical in determining the process outcome . To identify critical processing inputs in large databases, we have explored the use of Decision Tree Analysis with the decision metrics of Gain (i.e., Shannon Entropy changes), Gain Ratio, and a multiple hypergeometric distribution . The usefulness of this approach lies in its ability to treat "categorical" variables, which are typical of archived fermentation databases, as well as "continuous" variables . In this work, we demonstrate the use of Decision Tree Analysis for the problem of optimizing recombinant green fluorescent protein production in E . coli . A database of 85 fermentations was generated to examine the effect of 15 process input parameters on final biomass yield, maximum recombinant protein concentration, and productivity . The use of Decision Tree Analysis led to a considerable reduction in the fermentation database through the identification of the significant as well as insignificant inputs . However, different decision metrics selected different inputs and different numbers of inputs to classify the data for each output. Biotechnol Prog, 2002 Nov-Dec, 18(6), 1356 - 65 Genetic programming assisted stochastic optimization strategies for optimization of glucose to gluconic acid fermentation; Cheema JJ et al.; This article presents two hybrid strategies for the modeling and optimization of the glucose to gluconic acid batch bioprocess . In the hybrid approaches, first a novel artificial intelligence formalism, namely, genetic programming (GP), is used to develop a process model solely from the historic process input-output data . In the next step, the input space of the GP-based model, representing process operating conditions, is optimized using two stochastic optimization (SO) formalisms, viz., genetic algorithms (GAs) and simultaneous perturbation stochastic approximation (SPSA) . These SO formalisms possess certain unique advantages over the commonly used gradient-based optimization techniques . The principal advantage of the GP-GA and GP-SPSA hybrid techniques is that process modeling and optimization can be performed exclusively from the process input-output data without invoking the detailed knowledge of the process phenomenology . The GP-GA and GP-SPSA techniques have been employed for modeling and optimization of the glucose to gluconic acid bioprocess, and the optimized process operating conditions obtained thereby have been compared with those obtained using two other hybrid modeling-optimization paradigms integrating artificial neural networks (ANNs) and GA/SPSA formalisms . Finally, the overall optimized operating conditions given by the GP-GA method, when verified experimentally resulted in a significant improvement in the gluconic acid yield . The hybrid strategies presented here are generic in nature and can be employed for modeling and optimization of a wide variety of batch and continuous bioprocesses. Biotechnol Prog, 2002 Nov-Dec, 18(6), 1141 - 8 Modulation of phosphoenolpyruvate synthase expression increases shikimate pathway product yields in E . coli; Yi J et al.; Product yields in microbial synthesis are ultimately limited by the mechanism utilized for glucose transport . Altered expression of phosphoenolpyruvate synthase was examined as a method for circumventing these limits . Escherichia coli KL3/pJY1.216A was cultured under fed-batch fermentor conditions where glucose was the only source of carbon for the formation of microbial biomass and the synthesis of product 3-dehydroshikimic acid . Shikimate pathway byproducts 3-deoxy-D-arabino-heptulosonic acid, 3-dehydroquinic acid, and gallic acid were also generated . An optimal expression level of phosphoenolpyruvate synthase was identified, which did not correspond to the highest expression levels of this enzyme, where the total yield of 3-dehydroshikimic acid and shikimate pathway byproducts synthesized from glucose was 51% (mol/mol) . For comparison, the theoretical maximum yield is 43% (mol/mol) for synthesis of 3-dehydroshikimic acid and shikimate pathway byproducts from glucose in lieu of amplified expression of phosphoenolpyruvate synthase. Appl Microbiol Biotechnol, 2002 Dec, 60(4), 455 - 60 Epub 2002 Oct 15. Use of a histone H4 promoter to drive the expression of homologous and heterologous proteins by Penicillium funiculosum; Belshaw NJ et al.; Two genes encoding histone H4 (H4.1 and H4.2) from Penicillium funiculosum have been cloned and characterised . Structurally, the histone H4.1 gene is divergently linked to the histone H3 gene and the two genes are separated by approximately 800 bp . The transcription of the histone H4.1 and H4.2 genes in P . funiculosum appears to be distinctively regulated . Histone H4.1 mRNA showed a high steady-state level during the early stages of batch culture that decreased as growth reached the stationary phase . In contrast, the expression of the histone H4.2 gene was lower than that of H4.1 throughout batch growth and increased gradually with time . In order to expand the industrial application of P . funiculosum as a host for the production of heterologous proteins, the promoter of the histone H4.1 gene was successfully used to drive the expression of an intracellular bacterial enzyme, beta-glucuronidase, and a secreted homologous enzyme, xylanase C . The constitutive secretion of xylanase C was achieved in the absence of other xylanases by batch fermentation in the presence of glucose. Environ Technol, 2002 Oct, 23(10), 1147 - 56 Modelling of an activated primary settling tank including the fermentation process and VFA elutriation; Ribes J et al.; A complete model of a primary settler including both sedimentation and biological processes is presented . It is a one-dimensional model based on the solids flux concept and the conservation of mass that uses the Takacs model for the settling velocity, which is corrected by a compression function in the lower layers . The biological model is based on the ASM2 and enlarged with the fermentation model proposed by this research group . The settler was split in ten layers and the flux terms in the mass balance for each layer is obtained by means of the settling model . A pilot plant has been operated to study the primary sludge fermentation and volatile fatty acids (VFA) elutriation in a primary settler tank . The model has been tested with pilot plant experimental data with very good results . It has been able to simulate the VFA production in the settler and their elutriation with the influent wastewater for all the studied experiments . The developed model is easily applicable to secondary settlers and thickeners, also taking into account biological activity inside them. Nat Prod Lett, 2002 Dec, 16(6), 377 - 82 Microbial transformation of (+)-androsta-1 ,4-diene-3,17-dione by Cephalosporium aphidicola; Choudhary MI et al.; Fermentation of (+)-androsta-1,4-diene-3,17-dione ({structure: see text}) with Cephalosporium aphidicola for 8 days yielded oxidative and reductive metabolites, androst-4-ene-3,17-dione ({structure: see text}), 17beta-hydroxyandrosta-1,4-diene-3-one ({structure: see text}), 11alpha-hydroxyandrosta-1,4-diene-3,17-dione ({structure: see text}), 11alpha-hydroxyandrost-4-ene-3,17-dione ({structure: see text}), 11alpha,17beta-dihydroxyandrost-4-ene-3-one ({structure: see text}) and 11alpha,17beta-dihydroxyandrosta-1,4-diene-3-one ({structure: see text}) . The fermentation of {structure: see text} with Fusarium lini also yielded metabolites {structure: see text} . The structures of these metabolites were elucidated on the basis of spectroscopic techniques. J Anim Sci, 2002 Nov, 80(11), 3016 - 20 Effect of exogenous fibrolytic enzyme on ruminal fermentation and digestibility of alfalfa and rye-grass hay fed to lambs; Pinos-Rodriguez JM et al.; This experiment was carried out to study the effect of a directly fed exogenous fibrolytic enzyme on intake and digestion of DM, OM, protein, NDF, ADF, and hemicellulose of alfalfa and ryegrass hay by sheep . Four diets were randomly assigned to four ruminally cannulated lambs using a 4 x 4 Latin square design, repeated in time, with a factorial arrangement (2 x 2) of diets: 1) alfalfa hay; 2) alfalfa hay + exogenous fibrolytic enzymes (enzyme); 3) ryegrass hay; and 4) ryegrass hay + enzyme . Lambs consumed more DM and OM from alfalfa than from ryegrass hay (P < 0.001) . The ADF intake was not different between the hays, but NDF intake was lower for alfalfa (P < 0.001) . For both hays, the enzyme increased intake of DM (P < 0.01), as well as OM and CP (P < 0.05); however, NDF and ADF intake were not changed . Alfalfa hay had higher apparent digestibility of DM, OM, and CP (P < 0.001), but lower digestibility for NDF, ADF, and hemicellulose . The enzyme increased apparent digestibility of CP, hemicellulose (P < 0.05), and NDF (P < 0.10) for alfalfa . Also, for both hays, the enzyme improved N balance because lambs retained more N (P < 0.05) . The enzyme increased (P < 0.05) total VFA concentration (3 and 6 h) for both hays . Results from this trial indicate that directly fed exogenous fibrolytic enzymes may change ruminal fermentation, intake, and digestibility of forages with different nutritive value. J Anim Sci, 2002 Nov, 80(11), 3005 - 15 Effects of virginiamycin and monensin plus tylosin on ruminal protein metabolism in steers fed corn-based finishing diets with or without wet corn gluten feed; Ives SE et al.; Six ruminally cannulated steers (345 +/- 20 kg initial BW) were used in a 6 x 6 Latin square to evaluate effects of diet and antibiotics on ruminal protein metabolism . Two diets and three antibiotic treatments were arranged factorially . One diet contained (DM basis) 72% dry-rolled corn, 12% soybean meal, 10% alfalfa hay, and 4% molasses (SBM), and the other contained 63% dry-rolled corn, 30% wet corn gluten feed, and 5% alfalfa hay (WCGF) . Antibiotic treatments included control, virginiamycin (175 mg/d; VM), and monensin/tylosin (250 and 100 mg/d, respectively; MT) . Steers were fed at 12-h intervals at a rate of 2.4% of empty BW daily . Each period included 18 d of adaptation and 3 d of ruminal fluid collections . Samples were collected at 0, 2, 4, 6, 8, and 10 h after the morning feeding on d 19 and 20 . On d 21, rumens were dosed 2 h after the morning feeding with 350 g of solubilized casein to evaluate in vivo ruminal protease and deaminase activities . Ruminal fluid samples were collected 1, 2, 3, 4, and 6 h after the casein dose . On d 19 and 20, antibiotics had no effect on ruminal pH or concentrations of VFA, lactate, ammonia, ciliated protozoa, alpha-amino nitrogen (AAN), or peptide N, but VM reduced (P < 0.01) the concentration of isovalerate compared to MT and control . After casein dosing (d 21), peptide N concentration was unaffected by antibiotics, but AAN were higher (P < 0.01) for VM than MT and control . Relative to MT and control, VM reduced ruminal isovalerate (P = 0.05) and increased ruminal propionate (P < 0.01) on d 21 . Ruminal pH was lower (P < 0.01) in steers fed SBM than in steers fed WCGF, but lactate concentrations were unaffected by diet . Steers fed SBM had higher (P < 0.05) ruminal concentrations of total VFA and propionate . Ammonia concentrations were lower before feeding and higher after feeding for steers fed WCGF (P < 0.01) . Steers fed WCGF had higher counts of total ciliated protozoa than steers fed SBM (P < 0.05) due to greater Entodinium sp . (P < 0.05) . Steers fed WCGF had higher (P < 0.01) ruminal AAN and peptide N concentrations than those fed SBM on d 19 and 20 . After casein dosing, ruminal peptide N concentrations were similar, but AAN were lower (P < 0.01) for WCGF than SBM . Overall, VM appeared to depress ruminal deaminase activity, and MT had minimal effects on ruminal fermentation products . The protein in WCGF appeared to be more readily degradable than that in SBM. J Anim Sci, 2002 Nov, 80(11), 2978 - 88 Influence of rumen protein degradability and supplementation frequency on steers consuming low-quality forage: II . Ruminal fermentation characteristics; Bohnert DW et al.; Seven ruminally and duodenally cannulated steers (264 +/- 8 kg BW) consuming low-quality forage (5% CP; 61% NDF; 31% ADF) were used to determine the influence of CP degradability and supplementation frequency (SF) on ruminal fermentation characteristics . Treatments included an unsupplemented control and degradable intake protein (DIP) or undegradable intake protein (UIP) provided daily, every 3 d, or every 6 d . The DIP treatments (18% UIP) were calculated to provide 100% of the DIP requirement, while the UIP treatments (60% UIP) were provided on an isonitrogenous basis compared with DIP . Ruminal NH3-N was increased on the day all supplements were provided with supplemental CP (P = 0.04) and for DIP compared with UIP (P < 0.01) . Also, because ruminal NH3-N increased at a greater rate with DIP compared with UIP as SF decreased, a linear effect of SF x CP degradability interaction (P = 0.02) was observed . In addition, NH3-N was greater on the day only daily supplements were provided for supplemented treatments (P = 0.04), and decreased linearly (P < 0.01) as SF decreased . Concentration of total VFA increased linearly (P = 0.02) as SF decreased on the day all supplements were provided, whereas on the day only daily supplements were provided, total VFA were greater for UIP compared with DIP (P = 0.01), and decreased linearly (P < 0.01) as SF decreased . An interaction concerning the linear effect of SF and CP degradability (P = 0.02) was observed for ruminal liquid volume on the day all supplements were provided . This was the result of an increase in liquid volume with DIP as SF decreased compared with a minimal effect with UIP . In contrast, there was no influence of supplementation on liquid volume the day only daily supplements were provided . Ruminal liquid dilution rate was greater (P = 0.02) with CP supplementation on the day all supplements were provided . We did observe a quadratic effect of SF x CP degradability interaction (P = 0.01) for dilution rate because of a quadratic response with DIP (greatest value with the every-third-day treatment) compared with a decrease as SF decreased for UIP . On the day only daily supplements were provided, ruminal liquid dilution rate decreased linearly (P = 0.02) as SF decreased . These results suggest that DIP and UIP elicit different effects on ruminal fermentation when supplemented infrequently to ruminants consuming low-quality forage while not adversely affecting nutrient intake and digestibility. Lett Appl Microbiol, 2002, 35(6), 473 - 80 Investigating the prebiotic and gas-generating effects of selected carbohydrates on the human colonic microflora; Probert HM et al.; AIMS: To compare the fermentation of dietary carbohydrates with reference to their prebiotic and gas-generating capacity . METHODS AND RESULTS: Static anaerobic batch culture fermentations were carried out measuring gas generation and the prebiotic effect of five selected substrates (including various fructo-oligosaccharides, levan and maltodextrin) . The largest gas producer was levan, whilst those showing no significant difference to Actilight included oligofructose and maltodextrin . Gas composition data showed that hydrogen and carbon dioxide were the two most quantitatively important gases . The substrate that appeared to have the best prebiotic effect in vitro was branched chain fructo-oligosaccharide (FOS), followed by oligofructose, Actilight and maltodextrin which each exerted a similar effect . The substrate with the least bifidogenic effect was levan . CONCLUSIONS: The composition and total gas generation data showed that there was much variation between and within donor inocula . Generally, the lower gas producers had a more selective fermentation whilst larger gas producers were less specific . SIGNIFICANCE AND IMPACT OF THE STUDY: The study of these three parameters enabled a more complete picture of carbohydrate breakdown to be drawn and hence highlighted the need for potential prebiotics to be more extensively evaluated in order to reduce negative side-effects such as gas distension. Caries Res, 2002 Nov-Dec, 36(6), 417 - 22 Evaluation of the cariogenic potential of cassava flours from the Amazonian region; Rebelo Vieira JM et al.; Cassava flour is the main dietary carbohydrate source in the Amazonian region . The cariogenic potentials of the two main kinds of cassava flour - 'seca' and 'd'agua' - were evaluated in vivo and in vitro . A sweet made from a regional fruit (cupuacu) and a species of local fish (tambaqui) were used as positive and negative controls, respectively . For in vivo evaluation of dental plaque acidogenicity, the study had a crossover design, in four legs, with 19 volunteers in four treatment groups . Dental plaque pH was determined by the microtouch method before and for 60 min after food consumption . For in vitro evaluation, stimulated saliva of each volunteer was incubated with the food and pH variation was determined over 4 h . Carbohydrates were chemically determined in the flours and the majority was found to be water-insoluble . In vivo, plaque pH decreased significantly after the consumption of the sweet (p < 0.05), increased after the fish (p < 0.05), but did not change after intake of the flours (p > 0.05) . In vitro, the flours were slowly fermented by bacteria present in saliva . The in vivo and in vitro findings suggest that, in the form that the main cassava flours from the Amazonian region of Brazil are customarily eaten, they may be considered to have no or very low cariogenicity . J Microbiol Methods, 2003 Feb, 52(2), 239 - 44 Selective enrichment and purification of cultures of Methanosaeta spp; Janssen PH; A simple method for the isolation of axenic cultures of members of the obligately acetotrophic methanogenic genus Methanosaeta is described . To overcome the competitive advantage obtained by faster growing acetate-utilizing Methanosarcina spp . in batch enrichment cultures, acetone and isopropanol are used as the growth substrates for the enrichment step . Acetone- and isopropanol-utilizing bacteria slowly ferment these substrates to acetate, which allows Methanosaeta spp . to maintain the acetate concentration at levels below the threshold required for growth of Methanosarcina spp . These enrichments eventually develop dense populations of Methanosaeta spp., which can then be separated from contaminating microorganisms to yield axenic cultures . J Chromatogr A, 2002 Nov 29, 978(1-2), 153 - 64 Immobilized metal-ion affinity chromatography of recombinant Fab protein OPG C11 in the presence of EDTA-Mg(II); Xiang H et al.; Undesired adsorption of host cell proteins poses a big challenge for immobilized metal-ion affinity chromatography (IMAC) purification . In this study, by using His6-tagged protein Fab OPG C11 from Escherichia coli fermentation as a model, we found that the presence of low concentrations of EDTA-Mg2+ in feed streams weakens the adsorption but makes it more specific towards polyhistidine tag . By combining EDTA-Mg2+ treatment and periplasmic extraction, we developed a one-step purification procedure for His6-tagged recombinant Fab