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Evolution of Fusarium graminearum A3/5 grown in a glucose-limited chemostat culture at a slow dilution rate.

The evolution of Fusarium graminearum A3/5 grown in a glucose-limited chemostat at a dilution rate of 0.05 h-1 (doubling time of 13.9 h) was followed for 957 h or 69 generations. Periodic selection of advantageous mutants was monitored in the culture by determining increases and decreases in the concentration of cycloheximide-resistant macroconidia in the population. Six peaks in the concentration of cycloheximide-resistant macroconidia were observed representing five adaptive changes in the population; on average, an adaptive change occurred once every 148 +/- 22 h (mean +/- SE). The selection coefficient of strains present at the start of each increase in the concentration of cycloheximide-resistant macroconidia (i.e. after the establishment of a new advantageous strain) was determined relative to A3/5 and was found to increase progressively with time. When grown at a dilution rate of 0.05 h-1, the strain (A28-S) isolated from the last adaptive peak had a selection coefficient of 0.023 h-1 relative to A3/5, but A28-S lost its selective advantage when grown at a dilution rate of about 0.11 h-1 and was at a selective disadvantage when grown at a dilution rate higher than 0.11 h-1. The Km value (12 +/- 5 microM) for uptake of glucose by A28-S was significantly lower than that for A3/5. The spontaneous mutation rate from cycloheximide sensitivity to cycloheximide resistance was estimated to be 1.8 (+/- 0.2) x 10(-6) h-1 or 2.5 x 10(-5) generation.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological

Trophic relationships between Saccharomyces cerevisiae and Lactobacillus plantarum and their metabolism of glucose and citrate.

Glucose and citrate are two major carbon sources in fruits or fruit juices such as orange juice. Their metabolism and the microorganisms involved in their degradation were studied by inoculating with an aliquot of fermented orange juice a synthetic model medium containing glucose and citrate. At pH 3.6, their degradation led, first, to the formation of ethanol due to the activity of yeasts fermenting glucose and, eventually, to the formation of acetate resulting from the activity of lactobacilli. The yeast population always outcompeted the lactobacilli even when the fermented orange juice used as inoculum was mixed with fermented beet leaves containing a wider variety of lactic acid bacteria. The evolution of the medium remained similar between pH 3.3 and 5.0. At pH 3.0 or below, the fermentation of citrate was totally inhibited. Saccharomyces cerevisiae and Lactobacillus plantarum were identified as the only dominant microorganisms. The evolution of the model medium with the complex microbial community was successfully reconstituted with a defined coculture of S. cerevisiae and L. plantarum. The study of the fermentation of the defined model medium with a reconstituted microbial community allows us to better understand the behavior not only of fermented orange juice but also of many other fruit fermentations utilized for the production of alcoholic beverages.

Citrates

[Control of the fermentation activity during ensilage of Lolium italicum A. Br. and Medicago sativa L].

Samples with different dry matter contents of Lolium italicum A. Br., Medicago sativa L., have been ensiled in hermetically sealed containers, in order to study the evolution of microflora and its activity. The principal microbial groups (coliforms, proteolytics , lactic acid bacteria, clostridia, yeasts) have been detected and enumerated, in anaerobic atmosphere, after ensiling periods of 3, 6, 10, 13, 20, 100 days. At the same time, the samples were chemically analyzed for the detection of: pH, moisture, ashes, total and ammoniacal nitrogen, total reducing sugars, lactic acid, volatile fatty acids, short chain alcohols. The relations found between chemical and microbiological data, show that a strong lactic fermentation is not enough for the inhibition of silage spoiling microorganisms, such as coliforms, yeasts, and (less) proteolytics . This phenomenon may be related to the "quality" of the lactic acid microflora, more than to the characteristics of the environment.

Animal Feed

Draft results of a workshop to develop guidelines for studies involving microbial incidence or populations in the oral cavity.

The following five outlines are the results to date of the Workshop held in Rockville, Maryland, in January, 1990. The topics considered in these outlines are: (1) validation of immunological and/or nucleic acid identification probes, (2) cross-calibration of methods and/or laboratories for multi-laboratory cooperative studies, (3) choosing methods for identifying or describing microbial populations appropriate to the scientific question asked, (4) microbial ecology methods (e.g., population dynamics) for the oral cavity studies, and (5) epidemiological methods (e.g., incidence, risk factor analysis) for oral microbial studies. Each topic was considered by two independent groups of participants and later rationalized into one. These outlines are meant to be working outlines for evolution of a set of guidelines to advise on designing studies with microbial incidence and/or population components. We are publishing this preliminary version to elicit comment and criticism from people who did not attend the Workshop. (Attendance at the Workshop was necessarily limited by both space and funds). Some of the topic outlines have been condensed to save Journal space. The full document is available on request. The next stage will be an open forum to gather and discuss further amplification of the "Guidelines", planned for April 17, 1991, Acapulco, Mexico, in conjunction with the IADR/AADR Meeting. Written comments and requests for further information should be sent to the Workshop organizer (MIK) at the above address.(ABSTRACT TRUNCATED AT 250 WORDS)

DNA, Bacterial

Host-driven evolution shapes the polysaccharide utilization profiles of alga-associated Flavobacteriaceae.

BACKGROUND: Marine algae represent major producers of complex polysaccharides and serve as hosts for diverse microbial communities in the phycosphere. Flavobacteriaceae are among the key bacterial taxa involved in polysaccharide degradation and carbon remineralization in this environment. However, the extent to which algal hosts drive the divergence of polysaccharide utilization profiles in these bacteria remains unclear. RESULTS: We conducted a genome-resolved analysis of 103 cultured Flavobacteriaceae strains isolated from red, green, and brown macroalgae, as well as from diatoms and dinoflagellates. We found that macroalga-associated strains generally harbored more abundant and diverse CAZyme-encoding genes than their microalga-associated counterparts. Moreover, strains associated with different algal phyla showed distinct metabolic specializations that aligned with the typical polysaccharides of their respective hosts, strongly supporting host-specific adaptation. In four widely distributed genera (Maribacter, Flagellimonas, Polaribacter, Winogradskyella), CAZyme profile dissimilarity and key glycoside hydrolase gene divergence exhibited phylogenetic congruence with algal host phylogeny (Mantel r up to 0.76 and 0.85, respectively), indicative of host-associated functional adaptation. Using Maribacter as a model, cultivation experiments and transcriptome characterization demonstrated that polysaccharide utilization efficiency is not solely linked to the organization of genes into polysaccharide utilization loci (PULs), but also associated with the expression dynamics of key transcription factors (TFs), particularly those from AraC and DeoR families, whose expression patterns were coordinated with laminarin degradation. Notably, these two TF families also exhibited host-associated divergence patterns similar to those of CAZyme-encoding genes. Furthermore, analysis of the Tara Oceans metagenomic data indicated that, within the AraC and DeoR families, a higher proportion of genes were positively correlated with chlorophyll a content compared to other TF families, reinforcing their specialized roles in alga-associated bacterial lifestyles. CONCLUSIONS: Our integrative genomic and transcriptomic analyses reveal evolutionary and regulatory adaptation of marine Flavobacteriaceae to distinct algal hosts. These findings highlight algae-derived habitats as specialized niches that shape microbial metabolic potential, and suggest that carbohydrate metabolism plays a key role in host-driven bacterial evolution across global oceans. Video Abstract.

Flavobacteriaceae

Concurrent ecological and evolutionary processes contribute to mutualism breakdown between legumes and rhizobia.

Though they jointly shape community responses to environmental perturbations, ecology and evolution are often examined separately, even in microorganisms where both occur over short timescales. Here we examine ecological and evolutionary responses to 33 years of nitrogen fertilization using the legume-rhizobium mutualism. Pairing a manipulative inoculation study with full-length 16S rRNA gene amplicon sequencing and structural equation modeling allows us to synthesize across biological scales: whole bacterial community, genus Rhizobium, Rhizobium ASVs, and symbiosis plasmids. Clover's preferred partner decreases in N-addition soils, limiting host growth, while a diverse and largely uncharacterized Rhizobium community increases. This ecological change is compounded by a concurrent evolutionary degradation of symbiont partner quality via changing frequencies of symbiotic plasmids. Ecological (rarer symbionts) and evolutionary (inferior symbionts) processes each accounted for roughly half of this loss of host benefit, revealing that ecology and evolution jointly shape mutualism breakdown over the short timescales typical of microbial systems.

ecology

Somatic cell parasitism and the evolution of somatic tissue compatibility.

Selection pressures proposed to account for the convergent evolution of self/not-self recognition systems in lower organisms include defense against microbial parasites and somatic cell variants. Direct support for the existence of somatic cell parasites in natural populations has been lacking. I here report the occurrence of a somatic cell parasite in the cellular slime mold Dictyostelium mucoroides and discuss the implications of this phenomenon to the evolution of mechanisms of somatic tissue compatibility.

Animals

Of microbes and macrophages: entry, survival and persistence.

The macrophage is capable of fulfilling the roles of both host cell and effector cell in the immune clearance of many microbial infections. This dual capacity has led to the evolution of intriguing mechanisms whereby pathogens ensure the establishment and maintenance of intracellular infections. The strategies developed range from passive avoidance mechanisms to extremely active modulation of cell function. Recent advances have been made in our appreciation of the processes of intramacrophage parasitization from initial ligand-receptor interactions, through establishment of a stable intracellular environment, to the maintenance of a persistent infection.

Animals

Human lysosomal cathepsin G and granzyme B share a functionally conserved broad spectrum antibacterial peptide.

Human neutrophil lysosomal cathepsin G (cat G) exerts broad-spectrum antibacterial action in vitro against Gram-negative and -positive bacteria independent of its serine protease activity. We recently determined that an internal peptide of cat G (HPQYNQR), obtained after digestion of cat G with clostripain, possessed broad-spectrum antibacterial action in vitro, displaying an ED50 of 5 x 10(-5) M. In order to evaluate the structure-antibacterial properties of this peptide, synthetic variants with single alanine substitutions at each position were prepared and tested for antibacterial action. We found that alanine substitution for His-1 or Tyr-4, or certain modifications of the His-1 side chain, produced nonbactericidal peptides. A hexapeptide lacking the COOH-terminal Arg-7 but not a pentapeptide lacking both Gln-6 and Arg-7 possessed in vitro bactericidal activity. Interestingly, the cat G bactericidal peptide displays similarity to sequences within other serine proteases, notably the proposed cytotoxic granzymes present in the cytolytic granules of human and mouse cytotoxic T lymphocytes. We now report that an internal peptide of one human granzyme (granzyme B) with the sequence of HPAYNPK also displays bactericidal action in vitro. Our results suggest that an internal antibacterial domain among human serine proteases cat G and granzyme B has been functionally conserved through evolution perhaps for the purpose of host defense against microbial pathogens and targets of cytotoxic T lymphocyte killing.

Amino Acid Sequence

Effects of combinations of simulated acid rain and cadmium or zinc on microbial activity in soil.

There was little effect on the rate of CO2 evolution from glucose-supplemented soil, adjusted to pH 3.2 with a 2:1 combination of H2SO4 and fuming HNO3, and concomitant additions of 100 or 1000 ppm Cd or of 1000 or 10,000 ppm Zn (as sulfates) were no more inhibitory than in soil untreated with acid. In soil adjusted to pH 2.8, the lag in CO2 evolution was increased by 1 day, and was extended further by the concomitant addition of 10,000 but not 1000 ppm Zn or of 1000 but not 100 ppm Cd. The growth of Aspergillus niger in soil acidified to pH levels of 3.6 to 4.2 was further reduced by the addition of either 100 or 250 ppm Cd or of 1000 ppm Zn.

Air Pollutants

Microbial life at high pressures.

Deep-sea environment have selected for the evolution of unusual bacterial extremophiles which are adapted to life at high pressures. This review briefly characterises barophilic bacteria; compares and contrasts high pressure effects on cellular and biochemical processes in both barosensitive and barophilic bacteria; and presents molecular and genetic approaches which have been used to examine the basis of high pressure sensitivity in terrestrial barosensitive bacteria, or to dissect barophilic processes in deep-sea bacteria.

Air Pressure

Microbial catabolism, the carbon cycle and environmental pollution.

The establishment of a carbon cycle was a necessary prerequisite for the evolution of higher forms of life. This could not have been achieved without the direct participation of oxygen gas in certain metabolic reactions. The controlled activation of oxygen is catalyzed by microbial oxygenases; in principle, activated oxygen is hazardous to all living forms but without it, the degradative segment of the carbon cycle could not operate. The degradation of aromatic compounds is not an esoteric activity of a few specialized microorganisms. It occurs continuously, accompanied by fixation and cycling of oxygen on a massive scale; but like other global biochemical processes it tends to be neglected in general biological curricula. However, knowledge of the scope and limitations of microbial catabolic enzymes is central to the development of rational approaches to many of society's environmental concerns.

Acinetobacter

Protein H--a bacterial surface protein with affinity for both immunoglobulin and fibronectin type III domains.

Several bacterial species express surface proteins with affinity for the constant region (Fc) of immunoglobulin (Ig) G. The biological consequences of the interaction with IgG are poorly understood but it has been demonstrated that genes encoding different IgG Fc-binding proteins have undergone convergent evolution, suggesting that these surface molecules are connected with essential microbial functions. One of the molecules, protein H, is present in some strains of Streptococcus pyogenes, the most significant streptococcal species in clinical medicine. In contrast to other Ig-binding bacterial proteins tested, protein H was found to interact also with the neural cell adhesion molecule (N-CAM), a eukaryotic cell surface glycoprotein mediating homo- and heterophilic cell-cell interactions. The affinity for the interaction between protein H and N-CAM was 1.6 x 10(8)/M and the binding site on protein H was mapped to the NH2-terminal 80 amino acid residues. N-CAM and IgG are both members of the Ig superfamily and analogous to N-CAM, IgG binds to the NH2-terminal part of protein H. However, the binding sites for the two proteins were found to be separate, an unexpected result which was explained by the observation that the fibronectin type III (FNIII) domains and not the Ig-like domains of N-CAM are responsible for the interaction with protein H. Thus, the binding of N-CAM to protein H was blocked with fibronectin but not with IgG. Moreover, apart from fibronectin itself and N-CAM, fragments of fibronectin and the matrix protein cytotactin/tenascin containing FNIII domains also showed affinity for protein H.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Proteins

Evolutionary relationships of the Bacillus licheniformis macrolide-lincosamide-streptogramin B resistance elements.

Naturally occurring erythromycin (Em) resistance was found in 11 of the 18 Bacillus licheniformis isolates tested but was absent from a wide variety of other Bacillus strains. The Em resistance elements confer inducible macrolide-lincosamide-streptogramin B (MLS) resistance and are related to ermD , an MLS resistance element previously cloned from the chromosome of B. licheniformis 749. The MLS sensitive B. licheniformis strains and the other sensitive Bacillus strains tested, lack sequences with detectable homology to ermD . The sensitive B. licheniformis strains do exhibit homology to sequences which flank ermD in B. licheniformis 749. The relative sizes of the homologous DNA fragments suggest that the sensitive strains are lacking a 3.6 kb segment which contains ermD . It is shown that ermD is homologous to chromosomal DNA from Streptomyces erythreus ATCC 11635, an Em producing organism. These observations suggest to us that MLS resistance may have arisen in the Streptomyces and spread to B. licheniformis, another gram positive bacterium found in soil. It is further proposed that ermD is or was located on a transposon-like element and has spread and evolved further to yield a variety of related Staphylococcal and Streptococcal MLS determinants.

Anti-Bacterial Agents