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Antibacterial activity of Xylopia pancheri Baill. (Annonaceae) leaf extract was investigated against susceptible Staphylococcus aureus, methicillin-resistant S. aureus (MRSA) and extended-spectrum-β-lactamase (ESBL)-producing Pseudomonas aeruginosa. Results showed that X. pancheri leaf extract displayed antibacterial activity against S. aureus with higher inhibitory effect on MRSA compared to control gentamycin antibiotic, and IC

International audience; Mostly studied as a zoonosis, leptospirosis is also an environment-borne infection and most human cases originate from soil or water contaminations. Yet, only few studies have been interested in the survival of pathogenic Leptospira in freshwater. In this study, water microcosms were designed to evaluate the survival and virulence of Leptospira spp. for 2 years. Four commercial bottled drinking waters and a non-ionized water, all previously filter-sterilized, were studied. Either one of two Leptospira interrogans strains, one Leptospira borgpetersenii strain, or a saprophytic Leptospira biflexa was inoculated in these waters under nutrient-deprived conditions. Molecular, microscopic and cultural approaches were used to study Leptospira survival. Direct virulence of the pathogens was assessed using animal challenge without re-culturing. Our results confirmed the capacity of pathogenic Leptospira to survive for more than a year in water. In addition, we showed the ability of L. interrogans in nutrient-deprived conditions to directly cause systemic infection in susceptible animals. To our knowledge, this is the first report of direct infection of a susceptible host with Leptospira following a long starvation and survival period in nutrient-deprived water. Our results also suggest that Leptospira turned into a physiological "survival" state in harsh freshwater conditions. These data are of prime importance considering that freshwater is a major source of Leptospira infections. Environmental survival and virulence of pathogenic Leptospira spp. are becoming a crucial challenge to determine the environmental risk and adopt relevant prevention and control strategies.

Plant culture integration within aquaculture activities is a topic of recent interest with economic and environmental benefits. Shrimp farming activities generate nutrient-rich waste trapped in the sediments of farming ponds or release in the mangrove area. Thus, we investigate if the halophytes species naturally growing around the pond can use nitrogen and carbon from shrimp farming for remediation purposes. Halophyte biomasses and sediments influenced by shrimp farm effluents, were collected in two farms in New-Caledonia. All samples were analyzed for their C and N stable isotopic composition and N content. Higher δ15N values were found in plants influenced by shrimp farm water thus evidenced their abilities to take nutrient derived from shrimp farming. Deep root species Chenopodium murale, Atriplex jubata, Suaeda australis and Enchylaena tomentosa appears more efficient for shrimp pond remediation. This work demonstrates that halophytes cultivation in shrimp ponds with sediments, could be effective for the pond's remediation.

The ecological restoration of nickel mining-degraded areas in New Caledonia is strongly limited by low availability of soil mineral nutrients, metal toxicity, and slow growth rates of native plant species. In order to improve plant growth for restoration programs, special attention was paid to interactions between plant and soil microorganisms. In this study, we evaluated the influence of inoculation with Curtobacterium citreum BE isolated from a New Caledonian ultramafic soil on arbuscular mycorrhizal symbiosis and growth of Tetraria comosa, an endemic sedge used in restoration programs. A greenhouse experiment on ultramafic substrate was conducted with an inoculum comprising two arbuscular mycorrhizal fungi (AMF) species isolated from New Caledonian ultramafic soils: Rhizophagus neocaledonicus and Claroideoglomus etunicatum. The effects on plant growth of the AMF and C. citreum BE inoculated separately were not significant, but their co-inoculation significantly enhanced the dry weight of T. comosa compared with the non-inoculated control. These differences were positively correlated with mycorrhizal colonization which was improved by C. citreum BE. Compared with the control, co-inoculated plants were characterized by better mineral nutrition, a higher Ca/Mg ratio, and lower metal translocation. However, for Ca/Mg ratio and metal translocation, there were no significant differences between the effects of AMF inoculation and co-inoculation.

In New Caledonia, one third of the main island is covered by ultramafic soils characterized by high levels of metals and low levels of plant nutrients. In these soils, metal tolerant bacteria may play a role in plant adaptation to the edaphic constraints. Recently, two new bacteria species belonging to the genus Burkholderia sensu lato were isolated from rhizospheric roots of pioneer plant growing in New Caledonian ultramafic soils and were able to tolerate high metal concentrations. The purpose of this study was to determine the major mechanisms by which these two bacteria were able to tolerate metals and if they were able to reduce metal absorption by plants from which they were isolated. Bioaccumulation, adsorption on cell surface, ability to form biofilm and production of exopolysaccharides were assessed. A focus was carried out on Ni, generally the most toxic metal in New Caledonian ultramafic soils. All experiments were performed in comparison with a reference strain isolated from a non-ultramafic habitat, Paraburkholderia caribensis MWAP64T. The two ultramafic bacterial species showed a high tolerance to Ni with a Ni–IC50 of 5 mM for Caballeronia novacaledonica STM10272T and 30 mM for Paraburkholderia ultramafica STM10279T, values significantly higher than those of P. caribensis MWAP64T (2 mM). The major mechanism involved in the bacterial Ni tolerance was related to biofilm formation for C. novacaledonica STM10272T and to acidic exopolysaccharide production for P. ultramafica STM10279T. Both species alleviated metal contents in roots of Tetraria comosa, an endemic pioneer herbaceous species growing on New Caledonian ultramafic soils.

The present study focused on the characterization of 10

The present study focused on the characterization of 10 Curtobacterium citreum strains isolated from the rhizosphere of pioneer plants growing on ultramafic soils from New Caledonia. Taxonomic status was investigated using a polyphasic approach. Three strains (BE, BB, and AM) were selected in terms of multiple-metal resistance and plant-growth-promoting traits. They were tested on sorghum growing on ultramafic soil and compared with the reference strain C. citreum DSM20528T. To better understand the bacterial mechanisms involved, biosorption, bioaccumulation, and biofilm formation were investigated for the representative strain of the ultramafic cluster (strain BE) versus C. citreum DSM20528T. The polyphasic approach confirmed that all native isolates belong to the same cluster and are C. citreum. The inoculation of sorghum with strains BE and BB significantly reduced Ni content in shoots compared with inoculation with C. citreum DSM20528T and control values. This result was related to the higher Ni tolerance of the ultramafic strains compared with C. citreum DSM20528T. Ni biosorption and bioaccumulation showed that BE exhibited a lower Ni content, which is explained by the ability of this strain to produce exopolysaccharides involved in Ni chelation. We suggested that ultramafic C. citreum strains are more adapted to this substrate than is C. citreum DSM20528T, and their features allow them to enhance plant metal tolerance.

The shallow submarine hydrothermal field of the Prony Bay (New Caledonia) discharges hydrogen- and methane-rich fluids with low salinity, temperature (

Despite their potential importance as analogs of primitive microbial metabolisms, the knowledge of the structure and functioning of the deep ecosystems associated with serpentinizing environments is hampered by the lack of accessibility to relevant systems. These hyperalkaline environments are depleted in dissolved inorganic carbon (DIC), making the carbon sources and assimilation pathways in the associated ecosystems highly enigmatic. The Prony Bay Hydrothermal Field (PHF) is an active serpentinization site where, similar to Lost City (Mid-Atlantic Ridge), high-pH fluids rich in H-2 and CH4 are discharged from carbonate chimneys at the seafloor, but in a shallower lagoonal environment. This study aimed to characterize the subsurface microbial ecology of this environment by focusing on the earliest stages of chimney construction, dominated by the discharge of hydrothermal fluids of subseafloor origin. By jointly examining the mineralogy and the microbial diversity of the conduits of juvenile edifices at the micrometric scale, we find a central role of uncultivated bacteria belonging to the Firmicutes in the ecology of the PHF. These bacteria, along with members of the phyla Acetothermia and Omnitrophica, are identified as the first chimneys inhabitants before archaeal Methanosarcinales. They are involved in the construction and early consolidation of the carbonate structures via organomineralization processes. Their predominance in the most juvenile and nascent hydrothermal chimneys, and their affiliation with environmental subsurface microorganisms, indicate that they are likely discharged with hydrothermal fluids from the subseafloor. They may thus be representative of endolithic serpentinization-based ecosystems, in an environment where DIC is limited. In contrast, heterotrophic and fermentative microorganisms may consume organic compounds from the abiotic by-products of serpentinization processes and/or from life in the deeper subsurface. We thus propose that the Firmicutes identified at PHF may have a versatile metabolism with the capability to use diverse organic compounds from biological or abiotic origin. From that perspective, this study sheds new light on the structure of deep microbial communities living at the energetic edge in serpentinites and may provide an alternative model of the earliest metabolisms.

While reef degradation is occurring worldwide, it is not uncommon to see phase shifts from coral to macroalgal dominated reefs. Numerous studies have addressed the mechanisms by which macroalgae may outcompete corals and a few recent studies highlighted the putative role of bacteria at the interface between macroalgae and corals. Some studies suggest that macroalgae may act as vectors and/or foster proliferation of microorganisms pathogenic for corals. Using a combination of high throughput sequencing, bacterial culturing, and in situ bioassays we question if the adversity of macroalgal associated bacteria to corals is mediated by specific bacterial taxa. Using Illumina sequencing, we characterized and compared the bacterial community from two Lobophora (Dictyotales, Phaeophyceae) species. The two species presented distinctive bacterial communities. Both species shared approximately half of their OTUs, mainly the most abundant bacteria. Species-specific OTUs belong to Planctomycetes, Proteobacteria, and Bacteroidetes. In total, 16 culturable bacterial strain were isolated and identified from the Lobophora surface, consisting of 10 genera (from nine families, four classes, and three phyla), some of which are not known as, but are related to pathogens involved in coral diseases, and others are naturally associated to corals. When patches of marine agar with 24 h cultures of each of these bacteria were placed in direct contact with the branches of the scleractinian coral Acropora muricata, they caused severe bleaching after 24 h exposure. Results suggest that regardless of taxonomic affinities, increase in density of these bacteria can be adverse to corals. Nevertheless, the microbial community associated to macroalgal surface may not represent a threat to corals, because the specific bacterial screening and control exerted by the alga preventing specific bacterial proliferation. UIVIR ENTROPIE (UR, IRD, CNRS); postdoctoral fellowship of Portuguese National Science Foundation (Fundacao para a Ciencia e a Tecnologia, FCT) [SFRH/BPD/63703/2009, SFRH/BPD/107878/2015]; FCT program [UlD/IVItilti/04326/2013] info:eu-repo/semantics/publishedVersion

This study investigated soluble (Sol-EPS), loosely bound (LB-EPS), and tightly bound extracellular polymeric substances (TB-EPS) harvested from biofilm and planktonic cultures of the marine bacterium Pseudoalteromonas ulvae TC14. The aim of the characterization (colorimetric methods, FTIR, GC-MS, NMR, HPGPC, and AFM analyses) was to identify new anti-biofilm compounds; activity was assessed using the BioFilm Ring Test®. A step-wise separation of EPS was designed, based on differences in water-solubility and acidity. An acidic fraction was isolated from TB-EPS, which strongly inhibited biofilm formation by marine bacterial strains in a concentration-dependent manner. The main constituents of this fraction were characterized as two glucan-like polysaccharides. An active poly(glutamyl-glutamate) fraction was also recovered from TB-EPS. The distribution of these key EPS components in Sol-EPS, LB-EPS, and TB-EPS was distinct and differed quantitatively in biofilm vs planktonic cultures. The anti-biofilm potential of the fractions emphasizes the putative antifouling role of EPS in the environment.

The taxonomic status of eleven rhizospheric bacterial strains belonging to the genus Burkholderia and isolated from roots of Costularia (Cyperaceae), tropical herbaceous pioneer plants growing on ultramafic soils in New Caledonia, was investigated using a polyphasic taxonomic approach. The genetic analyses (16S rRNA genes, gyrB, recA, nreB and cnr) confirmed that all strains are Burkholderia and cluster into two separated groups. The DNA hybridization results showed low relatedness values to the closest relatives Burkholderia species. The phenotypic analyses confirmed that the two groups of strains could be differentiated from each other and from other known Burkholderia species. This polyphasic study revealed that these two groups of strains represent each a novel species of Burkholderia, for which the names Burkholderia novacaledonica sp. nov. (type strain STM10272(T)=LMG28615(T)=CIP110887(T)) and B. ultramafica sp. nov. (type strain STM10279(T)=LMG28614(T)=CIP110886(T)) are proposed, respectively. These strains of Burkholderia presented specific ecological traits such as the tolerance to the extreme edaphic constraints of ultramafic soils: they grew at pH between 4 and 8 and tolerate the strong unbalanced Ca/Mg ratio (1/19) and the high concentrations of heavy metals i.e. Co, Cr, Mn and Ni. Noteworthy B. ultramafica tolerated nickel until 10mM and B. novacaledonica up to 5mM. The presence of the nickel (nreB) and cobalt/nickel (cnr) resistance determinants encoding for protein involved in metal tolerance was found in all strains of both groups. Moreover, most of the strains were able to produce plant growth promoting molecules (ACC, IAA, NH3 and siderophores). Such ecological traits suggest that these new species of Burkholderia might be environmentally adaptable plant-associated bacteria and beneficial to plants.

International audience; Active carbonate chimneys from the shallow marine serpentinizing Prony Hydrothermal Field were sampled 3 times over a 6 years period at site ST09. Archaeal and bacterial communities composition was investigated using PCR-based methods (clone libraries, Denaturating Gel Gradient Electrophoresis, quantitative PCR) targeting 16S rRNA genes, methyl coenzyme M reductase A and dissimilatory sulfite reductase subunit B genes. Methanosarcinales (Euryarchaeota) and Thaumarchaea were the main archaeal members. The Methanosarcinales, also observed by epifluorescent microscopy and FISH, consisted of two phylotypes that were previously solely detected in two other serpentinitzing ecosystems (The Cedars and Lost City Hydrothermal Field). Surprisingly, members of the hyperthermophilic order Thermococcales were also found which may indicate the presence of a hot subsurface biosphere. The bacterial community mainly consisted of Firmicutes, Chloroflexi, Alpha-, Gamma-, Beta-, and Delta-proteobacteria and of the candidate division NPL-UPA2. Members of these taxa were consistently found each year and may therefore represent a stable core of the indigenous bacterial community of the PHF chimneys. Firmicutes isolates representing new bacterial taxa were obtained by cultivation under anaerobic conditions. Our study revealed diverse microbial communities in PHF ST09 related to methane and sulfur compounds that share common populations with other terrestrial or submarine serpentinizing ecosystems.

A new species of oligoglucuronan was previously extracted from a polysaccharide crude fraction produced by a Sinorhizobium meliloti mutant strain, by chromatography on l -histidine immobilized onto poly(ethylene–vinyl alcohol) (PEVA) hollow-fiber membranes. Until now, only one specific species of oligoglucuronan was selectively adsorbed on the membranes with immobilized histidine. In this work, the specific membranes, commonly used to purify proteins, were tested on different non-ionic and anionic carbohydrate components. We determined only anionic oligosaccharides are selectively retained on these membranes. The affinity between the l -His–PEVA membranes and oligoglucuronans varying in the degree of polymerization and the degree of substitution by acetyl residues was studied.

Low-molecular-mass polysaccharides produced by Rhizobia strains may be composed of a mixture of cyclic (1→2)-β- d -glucans substitued by sn -1-phosphoglycerol, succinic acid or methylmalonic acid or unsubstituted, and of oligosaccharides produced de novo or by degradation of the high-molecular-mass polysaccharides. The Sinorhizobium meliloti M5N1CS mutant strain (NCIMB 40472) produces unsubstituted cyclic glucans and these substituted by sn -1-phosphoglycerol, a wide variety of oligoglucuronans and high-molecular-mass glucuronans partially acetylated at the C2 and/or the C3 position. Until now, the purification of oligoglucuronans with a specific acetylation degree was not possible. In this work, we have studied the possibility of using l -histidine immobilized onto poly(ethylene–vinyl alcohol) hollow-fiber membranes for the separation of acetylated oligoglucuronans. Membranes with immobilized histidine known for purification of proteins based on dipole interaction were studied for the selective adsorption of glucuronans with discrimination of acetylation.

The Mediterranean Sea has rarely been investigated for the characterization of marine bacteria as compared to other marine environments such as the Atlantic or Pacific Ocean. Bacteria recovered from inert surfaces are poorly studied in these environments, when it has been shown that the community structure of attached bacteria can be dissimilar from that of planktonic bacteria present in the water column. The objectives of this study were to identify and characterize marine bacteria isolated from biofilms developed on inert surfaces immersed in the Mediterranean Sea and to evaluate their capacity to form a biofilm in vitro. Here, 13 marine bacterial strains have been isolated from different supports immersed in seawater in the Bay of Toulon (France). Phylogenetic analysis and different biological and physico-chemical properties have been investigated. Among the 13 strains recovered, 8 different genera and 12 different species were identified including 2 isolates of a novel bacterial species that we named Persicivirga mediterranea and whose genus had never been isolated from the Mediterranean Sea. Shewanella sp. and Pseudoalteromonas sp. were the most preponderant genera recovered in our conditions. The phenotypical characterization revealed that one isolate belonging to the Polaribacter genus differed from all the other ones by its hydrophobic properties and poor ability to form biofilms in vitro. Identifying and characterizing species isolated from seawater including from Mediterranean ecosystems could be helpful for example, to understand some aspects of bacterial biodiversity and to further study the mechanisms of biofilm (and biofouling) development in conditions approaching those of the marine environment.

International audience; Biofilm formation is a key step during marine biofouling, the natural colonization of immersed substrata, leading to major economic and ecological consequences. Consequently, bacteria have been used for the screening of new non-toxic antifoulants: the adhesion of five strains isolated on three French locations was monitored using a fluorescence-based assay and toxicity was also evaluated. Nine biocides including commercial, natural and natural-derived products were tested. The commercial antifoulants, TBTO and Sea Nine showed low EC(50) but high toxicity. The non-commercial products TFA-Z showed significant anti-adhesion activities and appeared to be non-toxic, suggesting a specific anti-adhesion mechanism. In addition, the strains could be classified depending on their sensitivity to the molecules used even if strain sensitivity also depended on the molecules tested. In conclusion, TFA-Z would be a promising candidate as non-toxic antifoulant and our results strengthen the need to perform antifouling bioassays with a panel of strains showing different response profiles.

The extracytoplasmic function sigma factor AlgU of Pseudomonas aeruginosa is responsible for alginate overproduction, leading to mucoidy and chronic infections of cystic fibrosis patients. We investigated here the role of AlgU in the formation of nonmucoid biofilms. The algU mutant of P. aeruginosa PAO1 (PAOU) showed a dramatic impairment in biofilm formation under dynamic conditions. PAOU was defective both in cell attachment to glass and in development of robust, shear-resistant biofilms. This was explained by an impaired production of extracellular matrix, specifically of the exopolysaccharide Psl, as revealed by microscopy and enzyme-linked immunosorbent assay. Complementing the algU mutation with a plasmid-borne algU gene restored wild-type phenotypes. Compared with that in PAO1, expression of the psl operon was reduced in the PAOU strain, and the biofilm formation ability of this strain was partially restored by inducing the transcription of the psl operon. Furthermore, expression of the lectin-encoding lecA and lecB genes was reduced in the PAOU strain. In agreement with the requirement of LecB for type IV pilus biogenesis, PAOU displayed impaired twitching motility. Collectively, these genetic downregulation events explain the biofilm formation defect of the PAOU mutant. Promoter mapping indicated that AlgU is probably not directly responsible for transcription of the psl operon and the lec genes, but AlgU is involved in the expression of the ppyR gene, whose product was reported to positively control psl expression. Expressing the ppyR gene in PAOU partially restored the formation of robust biofilms.

The Rhizobium sp. T1 strain. which induces nodule formation on alfalfa and clover roots, produces, during growth, an extracellular polysaccharide composed of D-glucose and D-glucuronic acid noted glucoglucuronan. During the bacterial growth, the pH of the medium decreases slightly. The control of pH in the growth medium slightly reduces the glucoglucuronan production. Under the conditions tested in the present work, the weight-average molecular weight of the polymers produced with or without pH control are similar: Mw approximately 2 x 10(6); the repeating unit determined by chemical and NMR analyses corresponds to the disaccharide: --> 3)-beta-D-GlcpA-(1-->4)-beta-D-Glcp-(1 -->.

The extent of human contamination, via water and soil, makes leptospirosis an environmental zoonosis. However, the survival of pathogenic leptospires in the environment remains poorly studied. To date, the genus Leptospira is composed of Pathogens 1 (formerly pathogens) and 2 (former intermediates), Saprophytes 1 (formerly saprophytes) and 2 (new group of saprophytes). The reorganization undergone by the phylogeny of leptospires is based on the discovery of a great biodiversity of leptospires. There are now 64 species of which 46 have been isolated from the environment. The hypothesis of the soil as the environmental habitat of pathogenic leptospires, remobilized by water during rains, was confirmed thanks to a systematic review of the literature. In order to better understand the environmental way of life of leptospires, their survival was studied in water, soil solutions and soils. Thus, a survival of pathogenic leptospires of more than one year has been confirmed in nutrient-free water. In addition, the maintenance of their virulence after this long incubation has been demonstrated. On the other hand, soil solutions with neutral pH and low conductivity as well as soils with low cation exchange capacity and field capacity as well as low in sodium and calcium seem to promote the survival of pathogenic leptospires. Understand the environmental transmission of leptospirosis. Identifying the habitat of these pathogenic bacteria will improve control and prevention strategies for this potentially fatal disease, applicable to other environmental infections., L’importance des contaminations humaines, via les eaux et les sols, fait de la leptospirose une zoonose d’origine environnementale. Pourtant, la survie des leptospires pathogènes dans l’environnement reste peu étudiée.À ce jour, le genre Leptospira est composé des Pathogènes 1 (anciennement pathogènes) et 2 (anciens intermédiaires), Saprophytes 1 (anciennement saprophytes) et 2 (nouveau groupe de saprophytes). Le remaniement subi par la phylogénie des leptospires repose sur la découverte d’une grande biodiversité de leptospires. On dénombre aujourd’hui 64 espèces dont 46 ont été isolées depuis l’environnement. L’hypothèse du sol comme habitat environnemental des leptospires pathogènes, remobilisés par l’eau lors de pluies, a été confortée grâce à une revue systématique de la littérature. Afin de mieux comprendre le mode de vie environnemental des leptospires, leur survie a été étudiée en eaux, solutions de sols et sols. Ainsi, une survie de leptospires pathogènes de plus d’un an a été confirmée en eau dépourvue en nutriment. De plus, le maintien de leur virulence après cette longue incubation a été démontré. D’autre part, les solutions de sol à pH neutre et de faible conductivité ainsi que les sols à faible capacité d’échange cationique et capacité au champ ainsi que pauvres en sodium et calcium semblent favoriser la survie des leptospires pathogènes.Ces études contribuent à comprendre la transmission environnementale de la leptospirose. L’identification de l’habitat de ces bactéries pathogènes permettra d’améliorer les stratégies de contrôle et de prévention de cette maladie potentiellement mortelle, applicables à d’autres infections environnementales.

The zoonotic bacterium Leptospira interrogans is the aetiological agent of leptospirosis, a re-emerging infectious disease that is a growing public health concern. Most human cases of leptospirosis result from environmental infection. Biofilm formation and its contribution to the persistence of virulent leptospires in the environment or in the host have scarcely been addressed. Here, we examined spatial and time-domain changes in biofilm production by L. interrogans. Our observations showed that biofilm formation in L. interrogans is a highly dynamic process and leads to a polarized architecture. We notably found that the biofilm matrix is composed of extracellular DNA, which enhances the biofilm’s cohesiveness. By studying L. interrogans mutants with defective diguanylate cyclase and phosphodiesterase genes, we show that biofilm production is regulated by intracellular levels of bis-(3′–5′)-cyclic dimeric guanosine monophosphate (c-di-GMP) and underpins the bacterium’s ability to withstand a wide variety of simulated environmental stresses. Our present results show how the c-di-GMP pathway regulates biofilm formation by L. interrogans, provide insights into the environmental persistence of L. interrogans and, more generally, highlight leptospirosis as an environment-borne threat to human health.