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Effect of bacterial interactions on the spoilage of cold-smoked salmon.

Cold-smoked salmon is a lightly preserved fish product in which a mixed microbial flora develops during storage and where the interactive behaviour of micro-organisms may contribute to their growth and spoilage activity. The aim of this study was to assess the effect of the bacterial interactions between the main species contaminating the cold-smoked salmon on bacterial growth, chemical and sensory changes, and spoilage. First, Carnobacterium piscicola, Photobacterium phosphoreum, Lactobacillus sakei, Vibrio sp., Brochothrix thermosphacta and Serratia liquefaciens-like were inoculated as pure cultures on sterile cold-smoked salmon. All bacterial species grew well; Vibrio sp. was the fastest and L. sakei strains developed very rapidly as well with a high maximum cell density on cold-smoked salmon blocks (up to 10(9) cfu g(-1) after 10 days at 8 degrees C). Based on sensory analysis, Vibrio sp. was identified as non-spoilage bacteria, C. piscicola as very lightly and B. thermosphacta as lightly spoiling. L. sakei and S. liquefaciens-like were found to be the most spoiling bacteria. Secondly, C. piscicola and L. sakei, two species frequently occurring in the lactic flora of the product, were inoculated together and each of them in mixed cultures with respectively P. phosphoreum, Vibrio sp., B. thermosphacta, and S. liquefaciens-like. The growth of L. sakei was shown to strongly inhibit most of the co-inoculated strains i.e. P. phosphoreum, B. thermosphacta, S. liquefaciens-like and, to a lesser extent, Vibrio sp. The growth of C. piscicola seemed to be enhanced with B. thermosphacta and to develop earlier with P. phosphoreum and Vibrio sp. Conversely, S. liquefaciens-like and P. phosphoreum were weakly inhibited by C. piscicola. The main observation resulting from the sensory evaluation was the delay in the appearance of the spoilage characteristics in the mixed cultures with L. sakei, in particular L. sakei/ S. liquefaciens-like. On the other hand, the spoilage activity of the non-spoiler strains Vibrio sp. or the moderate spoilage strains B. thermosphacta and C. piscicola was increased when they were associated together. It is concluded that the spoilage behaviour of micro-organisms in mixed culture is significantly different from pure culture and explain the difficulty to find robust quality indices for this product.

Animals↗

Plant-microbe interactions and the new biotechnological methods of plant disease control.

Plants constitute an excellent ecosystem for microorganisms. The environmental conditions offered differ considerably between the highly variable aerial plant part and the more stable root system. Microbes interact with plant tissues and cells with different degrees of dependence. The most interesting from the microbial ecology point of view, however, are specific interactions developed by plant-beneficial (either non-symbiotic or symbiotic) and pathogenic microorganisms. Plants, like humans and other animals, also become sick, but they have evolved a sophisticated defense response against microbes, based on a combination of constitutive and inducible responses which can be localized or spread throughout plant organs and tissues. The response is mediated by several messenger molecules that activate pathogen-responsive genes coding for enzymes or antimicrobial compounds, and produces less sophisticated and specific compounds than immunoglobulins in animals. However, the response specifically detects intracellularly a type of protein of the pathogen based on a gene-for-gene interaction recognition system, triggering a biochemical attack and programmed cell death. Several implications for the management of plant diseases are derived from knowledge of the basis of the specificity of plant-bacteria interactions. New biotechnological products are currently being developed based on stimulation of the plant defense response, and on the use of plant-beneficial bacteria for biological control of plant diseases (biopesticides) and for plant growth promotion (biofertilizers).

Bacteria↗

Winemaking biochemistry and microbiology: current knowledge and future trends.

The fermentation of grape must and the production of premium quality wines are a complex biochemical process that involves the interactions of enzymes from many different microbial species, but mainly yeasts and lactic acid bacteria. Yeasts are predominant in wine and carry out the alcoholic fermentation, while lactic acid bacteria are responsible for malolactic fermentation. Moreover, several optional winemaking techniques involve the use of technical enzyme preparations. Considerable progress has been made recently in understanding the biochemistry and interactions of enzymes during the winemaking process. In this study, some of these recent contributions in the biochemistry of winemaking are reviewed. This article intends to provide an updated overview (including works published until December, 2003) on the main biochemical and microbiological contributions of the different techniques that can be used in winemaking. As well as considering the transformations that take place in traditional winemaking, the production of special wines, such as sparkling wines, 'sur lie' wines, and biologically aged wines, are also studied.

Carbonated Beverages↗

The influence of supragingival plaque control on clinical and microbial outcomes following the use of antibiotics for the treatment of periodontitis.

Although supragingival plaque control is essential to successful periodontal therapy, the role of plaque control following systemic antibiotic use in periodontal disease has not been well defined. This study evaluated, following antibiotic use, which clinical and microbial parameters appeared to be influenced primarily by the antibiotics, independent of plaque control, and which outcomes appeared to be dependent on plaque control. Two hundred thirty-six patients (236) with moderate to severe periodontitis were clinically evaluated and microbial samples were taken by their private-practice periodontists. All patients were treated with scaling and root planing and a variety of systemic antibiotics, which were selected based on the microbial and clinical profile of the patient. Three months after therapy, patients were reevaluated and grouped by post-treatment plaque control, as either having very good oral hygiene (LoPl: N = 143; < or = 10% plaque-covered surfaces) or poor oral hygiene (HiPl: N = 93; > or = 25% plaque-covered surfaces). The two groups had different plaque and bleeding scores initially, but similar numbers of pockets probing > 5 mm and similar microbial patterns. Although the LoPl group had a significantly greater reduction in plaque than the HiPl group, bleeding scores and probing depths changed comparably in both groups after antibiotic therapy. Plaque control influenced outcomes significantly, but in a complex manner. The LoPl group exhibited a significantly greater reduction in certain bacteria, for example P. gingivalis. Interactions between plaque control and specific microbial parameters significantly affected clinical outcomes, although neither alone was sufficient to predict outcomes following antibiotic therapy.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Interactions among microorganisms of the indigenous intestinal flora and their influence on the host.

The animal host and its intestinal microbial flora function together as a complex ecologic system in which there is a significant impact of the intestinal flora on the host as well as of components of the microbial flora on one another. Aerobic and anaerobic bacteria of the intestinal flora influence numerous anatomic, physiologic, and immunologic parameters of the host. Constituents of the indigenous intestinal flora also engage in a multitude of antagonistic and cooperative interactions. The normal bacterial intestinal flora represents an extremely important defense mechanism, which effectively interferes with the establishment of many important enteric pathogens. Mechanisms by which microorganisms suppress the growth of other microorganisms include modification of bile acids, stimulation of peristalsis, induction of immunologic responses, depletion of essential substrates from the environment, competition for attachment sites, creation of restrictive physiologic environments, and elaboration of antibiotic-like substances. Components of the intestinal microbial flora also interact synergistically in the induction of disease or the utilization of substrate.

Animals↗

Molecular cross talk between epithelial cells and pathogenic microorganisms.

The conference brought together epithelial cell biologists and molecular microbiologists and emphasized that these seemingly diverse disciplines are intricately intertwined. The model systems discussed throughout the meeting emphasized the novel approaches available to address key issues and begin to understand the molecular details of responses triggered at the microbial-epithelial interface. For example, co-crystallization of native ligand-receptor complexes as well as biologically or chemically altered forms of these complexes will allow fine details of receptor-ligand interactions to be determined. This approach is critical in development of new generation antimicrobial agents. Furthermore, transfection techniques that allow receptor expression in model epithelia, development of representative animal model systems, and development of transgenic mouse strains will aid in dissecting microbial-epithelial interactions and will provide further advances in studies on pathogenesis and tissue and host tropism. We are only beginning to uncover the nature of the bidirectional regulatory signals that occur between microbes and hosts. We know little about how these signals relate to the disease state, to microbial virulence, or to immune function. Clearly the cross talk between cell biologists and microbiologists is an important step in unraveling the events occurring between microbes and eukaryotic cells.

Animals↗

The effect of microbial phytase in a pearl millet-soybean meal diet on apparent digestibility and retention of nutrients, serum mineral concentration, and bone mineral density of nursery pigs.

Eighteen gilts (initial BW of 10.9 kg; age 4 wk) were used in a 35-d experiment to study the effects of adding microbial phytase to a pearl millet-soybean meal-based diet on growth, apparent digestibility and retention of nutrients, and bone mineral status. The dietary treatments were arranged in a 2 x 3 factorial with two levels of P (.58%, low-P or .95%, adequate-P) and three levels of microbial phytase (0, 700, or 1,000 units/kg of diet). Phosphorus, Ca, and N balance were determined from d 32 to 35. Blood samples were collected on d 0 and 35 for serum mineral analysis. Global bone mineral content (GBMC) and global bone mineral density (GBMD) were determined on d 0 and 35 using dual-energy x-ray absorptiometry (DXA). A microbial phytase level x P level interaction was observed for final BW, ADG, gain:feed (P < .001), and serum P concentration (P < .003). Phytase supplementation of the low-P diets increased final BW, ADG, gain: feed, and serum inorganic P concentration but did not improve growth or serum inorganic P concentration in pigs fed the adequate-P diets. The addition of microbial phytase to the low-P and adequate-P diets reduced fecal P (P < .01), fecal N (P < .05), increased P absorption and retention (P < .01), and increased N absorption (P < .05). Pigs fed the adequate-P diets absorbed more P (P < .01) but excreted more P (P < .01) in the feces. Phytase tended to increase Ca (P < .07) and N (P < .10) retention, apparent threonine digestibility (P < .06), and serum Zn concentration (P < .09). Phytase increased GBMC and GBMD (P < .04). Phosphorus also increased GBMC and GBMD (P < .007 and P < .002, respectively). These results show that supplemental microbial phytase in pearl millet-soybean meal diets increased P availability, decreased the amount of P excreted in feces, and increased bone mineralization in nursery pigs.

6-Phytase↗

Applications of pharmacogenomics in general dental practice.

The human oral cavity represents a complex ecology of approximately 500 microbial species existing as commensals, and interacting with human/host factors. Perturbation of this ecosystem can lead to diseases of the oral cavity. The oral cavity also acts as a mirror of complex systemic diseases. Unique challenges and opportunities in oral healthcare exist in the diagnosis and treatment of primary disorders of the oral cavity or manifestations of systemic diseases. The completion of human and microbial genome projects has provided a wealth of information that will permit the application of pharmacogenomics - how genetic variations within the commensal and the host will impact the efficacy of drugs, and the diagnosis and treatment of oral disease. This review highlights opportunities focusing on oral health where pharmacogenomics can be applied.

Diagnosis, Oral↗

Influence of precipitation and soil on transport of fecal enterococci in fractured limestone aquifers.

Limestone aquifers provide the main drinking water resources of southern Italy. The groundwater is often contaminated by fecal bacteria because of the interaction between rocks having high permeability and microbial pollutants introduced into the environment by grazing and/or manure spreading. The microbial contamination of springwater in picnic areas located in high mountains can cause gastrointestinal illness. This study was carried out in order to analyze the interaction between Enterococcus faecalis and the soil of a limestone aquifer and to verify the influence of this interaction on the time dependence of groundwater contamination. E. faecalis was chosen because, in the study area involved, it represents a better indicator than Escherichia coli. The research was carried out through field (springwater monitoring) and laboratory experiments (column tests with intact soil blocks). The transport of bacterial cells through soil samples was analyzed by simulating an infiltration event that was monitored in the study area. Comparison of laboratory results with data acquired in the field showed that discontinuous precipitation caused an intermittent migration of microorganisms through the soil and produced, together with dispersion in the fractured medium (unsaturated and saturated zones), an articulated breakthrough at the spring. The short distances of bacterial transport in the study area produced a significant daily variability of bacterial contamination at the field scale.

Calcium Carbonate↗

Molecular signals in the interactions between plants and microbes.

The field of plant-microbe interactions has witnessed several recent breakthroughs, such as the molecular details of vir gene induction, identification of Nod factors, and the cloning and characterization of avr genes. Other breakthroughs, such as the cloning and characterization of R genes, appear imminent. Parallels to mammalian systems are emerging in the world of plant-microbe interactions, for example, ion channels formed by Rhizobium proteins, similarities of hrp genes to pathogenicity genes of mammalian pathogens, and plant signal transduction via calcium and protein phosphorylation. We remain, however, largely ignorant of many facets of signaling in plant-microbe interactions. We know little about how microbial signals are perceived by plants or how subsequent signal transduction occurs within plant cells and are probably unaware of many of the microbe-generated signals to which plants respond or of plant-generated signals to which bacteria and fungi respond. Contributions from those working on the genetics, molecular biology, and physiology of bacteria, fungi, and plants will be required to address these questions. The many nonpathogenic plant-microbe interactions in addition to the Rhizobium-plant interaction remain relatively unexplored. Genetic and molecular approaches are being initiated to investigate the signaling that is likely to underlie interactions such as those between mycorrhizal fungi and plant roots and between epiphytic bacteria and plant leaf surfaces. The importance of these interactions to plant growth and development makes it likely that they will figure more prominently at future symposia.(ABSTRACT TRUNCATED AT 250 WORDS)

Plant Physiological Phenomena↗

Characterization of pKa values and titration shifts in the cytotoxic ribonuclease alpha-sarcin by NMR. Relationship between electrostatic interactions, structure, and catalytic function.

The electrostatic behavior of titrating groups in alpha-sarcin was investigated using 1H NMR spectroscopy. A total of 209 chemical shift titration curves corresponding to different protons in the molecule were determined over the pH range of 3.0-8.5. Nonlinear least-squares fits of the data to simple relationships derived from the Henderson-Hasselbalch equation led to the unambiguous determination of pKa values for all glutamic acid and histidine residues, as well as for the C-terminal carboxylate and most of the aspartic acids in the free enzyme. The ionization constants of catalytically relevant histidines, His50 and His137, and glutamic acid, Glu96, in the alpha-sarcin-2'-GMP complex were also determined. The pKa values of 15 ionizable groups (C-carboxylate, six aspartic acids, four glutamic acids, and four histidines) were found to be close to their normal values. On the other hand, a number of side chain groups, including those in the active center, showed pKa values far from their intrinsic values. Thus, the pKa values for active site residues His50, Glu96, and His137 were 7.7, 5.2, and 5.8 in the free enzyme and 7.6, approximately 4.8, and 6.8 in the alpha-sarcin-2'-GMP complex, respectively. The pKa values and the activity profile against ApA, as a function of pH, are in agreement with the proposed enzymatic mechanism (in common with RNase T1 and the family of the microbial ribonucleases), in which Glu96 and His137 act as a general base and general acid, respectively. In almost all microbial ribonucleases, a Phe-His interaction is present, which affects the pKa of one of the His residues at the active site (His137). The absence of this interaction in alpha-sarcin would explain the lower pKa value of this His residue, and provides an explanation for the decreased RNase activity of this protein as compared to those of other microbial ribonucleases.

Aspergillus↗

Synergistic interaction of catecholamine hormones and Mycobacterium avium results in the induction of interleukin-10 mRNA expression by murine peritoneal macrophages.

The results of this investigation provides evidence that catecholamine hormones interact with macrophages that are infected with Mycobacterium avium resulting in the induction of IL-10 mRNA and protein. The effect of catecholamine hormones was prevented by treating the cells with the beta-adrenergic receptor antagonist propranolol but not by alpha-adrenergic antagonist phentolamine. The effect of catecholamine stimulation was mimicked by the addition of beta-2 adrenergic agonists and by the addition of cAMP to the infected macrophage cultures. These observations suggest that sympathetic nervous system activation together with microbial infection results in a synergistic interaction that could result in the control of inflammatory processes.

Adrenergic alpha-Agonists↗

Affinity and kinetic analysis of the bovine plasma C-type lectin collectin-43 (CL-43) interacting with mannan.

Collectins are C-type lectins which have been implied to play an important role in the innate immune defence against microorganisms. The critical discriminatory event in the opsonization of microorganisms by collectins is the interaction of the C-type lectin domain with microbial carbohydrates. Surface plasmon resonance measurements allow for quantitative real-time measurements of binding interaction between immobilized carbohydrate and unlabelled lectin in solution. Binding analysis were carried out with purified collectin-43 (CL-43) which structurally is the simplest collectin consisting of only three polypeptides each terminating in a C-type lectin domain. The target was immobilized yeast mannan. The molecular mass of native CL-43 was determinated by mass spectroscopy to 99.8 kDa. The dissociation rate (kdiss) of the C-type lectin-carbohydrate binding was fast (1.19-1.36 x 10(-2) second-1), and the association rate (kass) was 4.37-5.07 x 10(5) M-1 second-1. The equilibrium constant for dissociation (Kd) was 2.68-2.72 x 10(-8) M.

Animals↗

In vitro interaction of flucytosine with conventional and new antifungals against Cryptococcus neoformans clinical isolates.

Combinations of flucytosine with conventional and new antifungals were evaluated in vitro against 30 clinical isolates of Cryptococcus neoformans. Synergy determined by checkerboard analysis was observed with combinations of fluconazole, itraconazole, voriconazole, amphotericin B, and caspofungin with flucytosine against 77, 60, 80, 77, and 67% of the isolates, respectively. Antagonism was never observed. Killing curves showed indifferent interactions between triazoles and flucytosine and synergy between amphotericin B and flucytosine.

Antifungal Agents↗

Antimicrobial substances and effects on sessile bacteria.

Biofilms occur in natural aquatic ecosystems and on surfaces of biomaterials. They are generally associated with clinical infections predominantly of prosthetic hip joints, heart valves and catheters. Sessile microorganisms may be intimately associated with each other and to solid substratum through binding to and inclusion into exopolymer matrices on biofilms. The establishment of functional colonies within the exopolymeric matrices generate physico-chemical gradients within biofilms, that modify the metabolism and cell-wall properties of the microorganism. A consequence of biofilm growth is an enhanced microbial resistance to chemical antimicrobial agents and antibiotics. Investigations on the antimicrobial efficacy of antibiotics, antiseptics and antimicrobial heavy ions, however, gave controversial results. No single antimicrobial substance has been developed for the efficient eradication of adherent bacteria. This review elucidates the mechanisms of microbial resistance in biofilms and strategies for the prevention of biofilm development. Pharmacokinetical and pharmacodynamical issues for the screening of biofilm-active drugs are presented. Combinations of antistaphylococcal antibiotics with rifampin may be advantageous for preventing and curing biomaterial infections.

Anti-Bacterial Agents↗

Effects of single D-amino acid substitutions on disruption of beta-sheet structure and hydrophobicity in cyclic 14-residue antimicrobial peptide analogs related to gramicidin S.

Gramicidin S (GS) is a 10-residue cyclic beta-sheet peptide with lytic activity against the membranes of both microbial and human cells, i.e. it possesses little to no biologic specificity for either cell type. Structure-activity studies of de novo-designed 14-residue cyclic peptides based on GS have previously shown that higher specificity against microbial membranes, i.e. a high therapeutic index (TI), can be achieved by the replacement of a single L-amino acid with its corresponding D-enantiomer [Kondejewski, L.H. et al. (1999) J. Biol. Chem. 274, 13181]. The diastereomer with a D-Lys substituted at position 4 caused the greatest improvement in specificity vs. other L to D substitutions within the cyclic 14-residue peptide GS14, through a combination of decreased peptide amphipathicity and disrupted beta-sheet structure in aqueous conditions [McInnes, C. et al. (2000) J. Biol. Chem. 275, 14287]. Based on this information, we have created a series of peptide diastereomers substituted only at position 4 by a D- or L-amino acid (Leu, Phe, Tyr, Asn, Lys, and achiral Gly). The amino acids chosen in this study represent a range of hydrophobicities/hydrophilicities as a subset of the 20 naturally occurring amino acids. While the D- and L-substitutions of Leu, Phe, and Tyr all resulted in strong hemolytic activity, the substitutions of hydrophilic D-amino acids D-Lys and D-Asn in GS14 at position 4 resulted in weaker hemolytic activity than in the L-diastereomers, which demonstrated strong hemolysis. All of the L-substitutions also resulted in poor antimicrobial activity and an extremely low TI, while the antimicrobial activity of the D-substituted peptides tended to improve based on the hydrophilicity of the residue. D-Lys was the most polar and most efficacious substitution, resulting in the highest TI. Interestingly, the hydrophobic D-amino acid substitutions had superior antimicrobial activity vs. the L-enantiomers although substitution of a hydrophobic D-amino acid increases the nonpolar face hydrophobicity. These results further support the role of hydrophobicity of the nonpolar face as a major influence on microbial specificity, but also highlights the importance of a disrupted beta-sheet structure on antimicrobial activity.

1-Naphthylamine↗

In vitro activity of six antibiotics against multiresistant staphylococci and other gram-positive cocci.

Sixty-two strains of Staphylococcus aureus and coagulase-negative staphylococci, 21 Streptococcus faecalis and 17 other strains of streptococci isolated from cases of endocarditis were tested for sensitivity against rifampicin, teicoplanin, vancomycin, fusidic acid, erythromycin and novobiocin. Only rifampicin, novobiocin and teicoplanin were found to be active against the great majority of these strains. The microbial properties of these antibiotics suggest the necessity of combinations for effective therapy. The combinations rifampicin + novobiocin and rifampicin + teicoplanin were additive and suppressed the emergence of resistant mutants. Thus according to in vitro tests, either of these two combinations would be suitable for prophylactic use in high-risk patients, especially those scheduled to receive prosthetic implants.

Anti-Bacterial Agents↗