Role of N-acetylglucosaminidase and N-acetylmuramidase activities in Enterococcus faecalis peptidoglycan metabolism

Identification of the full complement of peptidoglycan hydrolases detected by zymogram in Enterococcus faecalis extracts led to the characterization of two novel hydrolases that we named AtlB and AtlC. Both enzymes have a similar modular organization comprising a central catalytic domain fused to two LysM peptidoglycan-binding modules. AtlB and AtlC displayed N-acetylmuramidase activity, as demonstrated by tandem mass spectrometry analyses of peptidoglycan fragments generated by the purified enzymes. The genes encoding AtlB and AtlC were deleted either alone or in combination with the gene encoding AtlA, a previously described N-acetylglucosaminidase. No autolytic activity was detected in the triple mutant indicating that AtlA, AtlB, and AtlC account for the major hydrolytic activities in E. faecalis. Analysis of cell size distribution by flow cytometry showed that deletion of atlA resulted in the formation of long chains. Thus, AtlA digests the septum and is required for cell separation after cell division. We found that AtlB could act as a surrogate for AtlA, although the enzyme was less efficient at septum digestion. Deletion of atlC had no impact on cell morphology. Labeling of the peptidoglycan with N-[14C]acetylglucosamine revealed an unusually slow turnover as compared with model organisms, almost completely dependent upon the combined activities of AtlA and AtlB. In contrast to atlA, the atlB and atlC genes are located in putative prophages. Because AtlB and AtlC were produced in the absence of cell lysis or production of phage progeny, these enzymes may have been hijacked by E. faecalis to contribute to peptidoglycan metabolism.     

Food choice and food competition among the three major primate species of French Guiana

The three most important primates of French Guiana are frugivorous species, but to different degrees. Ateles paniscus is almost exclusively frugivorous, Alouatta seniculus frugivorous-folivorous, and Cehus apellu partially insectivorous; thus, the overall specific diet spectra arc well separated. During the season when fruit production is minimal, fruit intake by all species is also minimal which results in the greatest diversities in diet and the lowest diet overlaps. Real competition for the common main food category, i.e. fruits, only occurs during the most favourable period. Nevertheless, some fruit characteristics (e.g. size…) separate the monkey species from each other in fruit choice and make the interspecific competition of little importance.     

The peptidoglycan of Mycobacterium abscessus is predominantly cross-linked by L,D-transpeptidases

Few therapeutic alternatives remain for the treatment of infections due to multiresistant Mycobacterium abscessus. Here we show that the peptidoglycans of the "rough" and "smooth" morphotypes contain predominantly 3→3 cross-links generated by l,d-transpeptidases, indicating that these enzymes are attractive targets for the development of efficient drugs.     

Secondary metabolites from plant-associated Pseudomonas are overproduced in biofilm

Plant rhizosphere soil houses complex microbial communities in which microorganisms are often involved in intraspecies as well as interspecies and inter-kingdom signalling networks. Some members of these networks can improve plant health thanks to an important diversity of bioactive secondary metabolites. In this competitive environment, the ability to form biofilms may provide major advantages to microorganisms. With the aim of highlighting the impact of bacterial lifestyle on secondary metabolites production, we performed a metabolomic analysis on four fluorescent Pseudomonas strains cultivated in planktonic and biofilm colony conditions. The untargeted metabolomic analysis led to the detection of hundreds of secondary metabolites in culture extracts. Comparison between biofilm and planktonic conditions showed that bacterial lifestyle is a key factor influencing Pseudomonas metabolome. More than 50% of the detected metabolites were differentially produced according to planktonic or biofilm lifestyles, with the four Pseudomonas strains overproducing several secondary metabolites in biofilm conditions. In parallel, metabolomic analysis associated with genomic prediction and a molecular networking approach enabled us to evaluate the impact of bacterial lifestyle on chemically identified secondary metabolites, more precisely involved in microbial interactions and plant-growth promotion. Notably, this work highlights the major effect of biofilm lifestyle on acyl-homoserine lactone and phenazine production in P. chlororaphis strains.     

Fruit characters as a basis of fruit choice and seed dispersal in a tropical forest vertebrate community

Summary Interactions between a large community of vertebrate frugivore-granivores (including 7 species of large canopy birds, 19 species of rodents, 7 species of ruminants, and 6 species of monkeys), and 122 fruit species they consume, were studied for a year in a tropical rainforest in Gabon.The results show how morphological characters of fruits are involved in the choice and partitioning of the available fruit spectrum among consumer taxa. Despite an outstanding lack of specificity between fruit and consumer species, consideration of simple morphological traits of fruits reveals broad character syndromes associated with different consumer taxa. Competition between distantly related taxa that feed at the same height is far more important than has been previously supposed. The results also suggest how fruit characters could have evolved under consumer pressure as a result of consumer roles as dispersers or seed predators. Our analyses of dispersal syndromes show that fruit species partitioning occurs more between mammal taxa than between mammals and birds. There is thus a bird-monkey syndrome and a ruminant-rodent-elephant syndrome. The bird-monkey syndrome includes fruit species on which there is no pre-dispersal seed predation. These fruits (berries and drupes) are brightly colored, have a succulent pulp or arillate seeds, and no protective seed cover. The ruminant-rodent-elephant syndrome includes species for which there is pre-dispersal predation. These fruits (all drupes) are large, dull-colored, and have a dry fibrous flesh and well-protected seeds.     

Gonad development and filleting yield of common carp Cyprinus carpio L. reared in ponds in Eastern France

The processing of 232 common carp Cyprinus carpio made it possible to study the correlation between gonad maturation and filleting yield. The study was undertaken between April and August 1992 and was carried out on 4-year-old carp cultured in extensive ponds. Morphological parameters were recorded and relations between total weight and length, biomass of fillet, and biomass of gonad were determined. The histology of male and female gonads was examined in relation to the gonadosomatic index (GSI: up to 16% for females and less than 11% for males) and condition indices (total K or somatic K1). Filleting yields (32–36% of the total weight) decreased for females just prior to spawning and corresponded to a decline of K1. A rematuration phenomenon was observed, although females weighing less than 1200g were still immature. We suggest avoiding the filleting of large females before spawning and suggest estimating the date of appearance of this stage by referring to the positive correlation established in this paper between the number of degree-days and total fish weight.     

Community structure of the frugivours-folivorous forest mammals of Gabon

The distribution of 66 spcies of mammalian primary consumers in a Gabon rain forest is analysed by broad categories of habitat, height, activity cycle, diet and body weight. Aa few species are restricted to waterside habita; most inhabit dry-land forest. Species are almost equally divided between the roles of arboreal and terrestrial, diurnal and nocturnal, but there are five times as many frugivores as folivores. Terrestriality, large body size, and folivory are correlated with mixed diurnal-nocturnal activity. Frugivores ahve a wider range of ecological roles than folivores. Different taxa have radiated in similare patterns, suggesting similar ecologcical constraints on the diversification of different families.     

Le comportement du Chevrotain africain,Hyemoschus aquaticus Ogilby (Artiodactyla,Ruminantia)

The purpose of this study was to test the influence of social isolation during infancy on learning behavior in rats. Rats, reared isolated (n = 55) or paired (n = 60) for a period of 50 days following weaning, were tested on different learning tasks in an operant conditioning chamber. Isolation rearing produced significantly better learning when the training period directly followed the different housing conditions. After a 28 day period of social isolation for the subjects raised paired during infancy no differences in learning between isolated and paired reared animals could be observed. The differences are discussed in relation to the effects of stimulus deprivation.     

Heterogeneity of Proteinase Inhibitors in the Water-Soluble Organic Matrix from the Oyster Nacre

We extracted proteinase inhibitors from the nacre of the oyster Pinctada margaritifera with water. Mixing the nacre powder with water for 20 h led to a water-soluble fraction [0.24% (wt/wt) of nacre]. After dialysis of the water-soluble matrix through 6- to 8-kDa and 0.5-kDa membranes, the proteinase inhibitors were divided into low and high molecular weight fractions that contained inhibitors of papain, bovine cathepsin B, and human cathepsin L. We studied the heterogeneity of the inhibitors after separating the low molecular weight fraction according to charge and hydrophobicity. After multistep purification, mass spectrometry analysis revealed that a potent inhibitory fraction contained several molecules. This observation demonstrates the difficulties encountered in attempting to isolate individual metabolites from the complex mixture of molecules present in nacre matrix. Interestingly, the low molecular weight fraction contained specific inhibitors that could discern between cathepsin B and cathepsin L. The nacre organic inhibitors were active against several cysteine proteinases, yet they were more specific in relation to serine proteinases, because only proteinase K was inhibited. These results demonstrate, for the first time, the presence of active proteinase inhibitors in the mollusc shell, and it is possible that these inhibitors may play a role in either protection of proteins involved in shell formation or in defense against parasites, or both.     

Novel mechanism of resistance to glycopeptide antibiotics in Enterococcus faecium

Glycopeptides and beta-lactams are the major antibiotics available for the treatment of infections due to Gram-positive bacteria. Emergence of cross-resistance to these drugs by a single mechanism has been considered as unlikely because they inhibit peptidoglycan polymerization by different mechanisms. The glycopeptides bind to the peptidyl-D-Ala(4)-D-Ala(5) extremity of peptidoglycan precursors and block by steric hindrance the essential glycosyltransferase and D,D-transpeptidase activities of the penicillin-binding proteins (PBPs). The beta-lactams are structural analogues of D-Ala(4)-D-Ala(5) and act as suicide substrates of the D,D-transpeptidase module of the PBPs. Here we have shown that bypass of the PBPs by the recently described beta-lactam-insensitive L,D-transpeptidase from Enterococcus faecium (Ldt(fm)) can lead to high level resistance to glycopeptides and beta-lactams. Cross-resistance was selected by glycopeptides alone or serially by beta-lactams and glycopeptides. In the corresponding mutants, UDP-MurNAc-pentapeptide was extensively converted to UDP-MurNAc-tetrapeptide following hydrolysis of D-Ala(5), thereby providing the substrate of Ldt(fm). Complete elimination of D-Ala(5), a residue essential for glycopeptide binding, was possible because Ldt(fm) uses the energy of the L-Lys(3)-D-Ala(4) peptide bond for cross-link formation in contrast to PBPs, which use the energy of the D-Ala(4)-D-Ala(5) bond. This novel mechanism of glycopeptide resistance was unrelated to the previously identified replacement of D-Ala(5) by D-Ser or D-lactate.