The Use of FTIR Spectroscopy as a Tool for the Seasonal Variation Analysis and for the Quality Control of Polysaccharides from Seaweeds.

Macroalgae are a potentially novel source of nutrition and biologically active molecules. Proliferative species such as Eucheuma denticulatum, Solieria chordalis (red algae) and Sargassum muticum (brown alga) constitute a huge biomass that can be exploited. In this study, we focus on the extraction of polysaccharides from these three macroalgae species and the characterization of cell wall polysaccharides such as carrageenans, fucoidans and alginates by Fourier Transform Infrared spectroscopy with Attenuated Reflectance Module (FTIR-ATR). The comparison of purified extracts with commercial solutions of fucoidans, alginates or carrageenans shows a strong similarity between the spectra. It demonstrates that the methods of extraction that have been used are also suitable purifying technics. Moreover, it validates infrared spectroscopy as a quick, simple and non-destructive method for the accurate analysis of polysaccharides. The FTIR technique applied to samples collected at different periods of the year allowed us to highlight differences in the composition of fucoidans, alginates and carrageenans. Different classes corresponding to the season can be distinguished by statistical multidimensionnal analysis (Principal Component Analysis) showing that the structure of algal polysaccharides, related to bioactivity, depends on the period of harvest. FTIR results showed that S. chordalis and E. denticulatum possess a dominant type of carrageenan called iota-carrageenan. This type of carrageenan is in the majority when the alga is at maturity in its development cycle. During its growth phase, iota-carrageenan precursors can be detected by FTIR spectra, enabling a better control of the extraction and an application of these compounds in various economic sectors. When the alga E. denticulatum is in its juvenile stage, we found traces of kappa-carrageenan and nu-carrageenan polysaccharides in some extracts.

Sulfated Polysaccharides from Seaweed Strandings as Renewable Source for Potential Antivirals against Herpes simplex Virus 1.

Herpes simplex virus 1 (HSV-1) remains a prominent health concern widespread all over the world. The increasing genital infections by HSV-1 that might facilitate acquisition and transmission of HIV-1, the cumulative evidence that HSV-1 promotes neurodegenerative disorders, and the emergence of drug resistance signify the need for new antiviral agents. In this study, the in vitro anti-herpetic activity of sulfated polysaccharides (SPs) extracted by enzyme or hot water from seaweeds collected in France and Mexico from stranding events, were evaluated. The anti-herpetic activity evaluation of the semi-refined-polysaccharides (sr-SPs) and different ion exchange purified fractions showed a wide range of antiviral activity. Among them, the sr-SPs from the Rhodophyta Halymenia floresii showed stronger activity EC50 0.68 µg/mL with SI 1470, without cytotoxicity. Further, the antiviral activity of the sr-SPs evaluated at different treatment schemes showed a high EC50 of 0.38 µg/mL during the viral adsorption assays when the polysaccharide and the virus were added simultaneously, whilst the protection on Vero cell during the post-infection assay was effective up to 1 h. The chemical composition, FTIR and 1H NMR spectroscopic, and molecular weights of the sr-SPs from H. floresii were determined and discussed based on the anti-herpetic activity. The potential utilization of seaweed stranding as a source of antiviral compounds is addressed.

Cinnamomum cassia and Syzygium aromaticum Essential Oils Reduce the Colonization of Salmonella Typhimurium in an In Vivo Infection Model Using Caenorhabditis elegans.

The regulation of intestinal colonization in livestock by means of non-bactericidal additives is an important management lever for zoonotic bacteria such as Salmonella spp. Caenorhabditis elegans is proposed here as a model for the evaluation of five essential oils (EOs) as anti-colonization products against Salmonella Typhimurium. An evaluation of the toxicity of EOs for C. elegans showed LD50 values ranging from 74.5 ± 9.6 µg/mL for Cinnamomum cassia (CEO) to 271.6 ± 14.9 µg/mL for Syzygium aromaticum (SyEO). Both EOs significantly inhibited bacterial colonization in the digestive tract of C. elegans with reductions of 0.88 and 0.70 log CFU/nematode at nontoxic concentrations of 50 µg/mL and 150 µg/mL, respectively. With the minimal bactericidal concentrations of CEO and SyEO against S. Typhimurium being 312.5 µg/mL and 625 µg/mL, respectively, an antibacterial effect can be excluded to explain the inhibition of the bacterial load. The anti-colonizing activity of these two EOs could, however, be related to an inhibition of the swimming motility, which was significantly reduced by 23.47% for CEO at 50 µg/mL and 19.56% for SyEO at 150 µg/mL. This study shows the potential of C. elegans as a predictive in vivo model of anti-colonizing activities that is suitable for the evaluation of essential oils.

Effects of Ulva sp. Extracts on the Growth, Biofilm Production, and Virulence of Skin Bacteria Microbiota: Staphylococcus aureus, Staphylococcus epidermidis, and Cutibacterium acnes Strains.

Ulva sp. is known to be a source of bioactive compounds such as ulvans, but to date, their biological activity on skin commensal and/or opportunistic pathogen bacteria has not been reported. In this study, the effects of poly- and oligosaccharide fractions produced by enzyme-assisted extraction and depolymerization were investigated, for the first time in vitro, on cutaneous bacteria: Staphylococcus aureus, Staphylococcus epidermidis, and Cutibacterium acnes. At 1000 µg/mL, poly- and oligosaccharide fractions did not affect the growth of the bacteria regarding their generation time. Polysaccharide Ulva sp. fractions at 1000 µg/mL did not alter the bacterial biofilm formation, while oligosaccharide fractions modified S. epidermidis and C. acnes biofilm structures. None of the fractions at 1000 µg/mL significantly modified the cytotoxic potential of S. epidermidis and S. aureus towards keratinocytes. However, poly- and oligosaccharide fractions at 1000 µg/mL induced a decrease in the inflammatory potential of both acneic and non-acneic C. acnes strains on keratinocytes of up to 39.8%; the strongest and most significant effect occurred when the bacteria were grown in the presence of polysaccharide fractions. Our research shows that poly- and oligosaccharide Ulva sp. fractions present notable biological activities on cutaneous bacteria, especially towards C. acnes acneic and non-acneic strains, which supports their potential use for dermo-cosmetic applications.

Poly- and Oligosaccharide Ulva sp. Fractions from Enzyme-Assisted Extraction Modulate the Metabolism of Extracellular Matrix in Human Skin Fibroblasts: Potential in Anti-Aging Dermo-Cosmetic Applications.

Ulva sp. is known to be a source of bioactive compounds such as ulvans, but their biological activity on human dermal fibroblast extracellular matrix (ECM) is poorly reported. In this work, the regulation of ECM has been investigated for the first time at both proteomic and transcriptomic levels in normal human skin dermal fibroblasts, after 48 h of incubation with poly- and oligosaccharide fractions from Ulva sp. obtained after enzyme-assisted extraction and depolymerization. Cell proliferation enhancement (up to +68%) without exhibiting any cytotoxic effect on fibroblasts was demonstrated at 50 and 1000 µg/mL by both fractions. At the proteomic level, polysaccharide fractions at 1000 µg/mL enhanced the most the synthesis of glycosaminoglycans (GAGs, up to +57%), total collagen, especially types I (up to +217%) and III, as well as the synthesis and activity of MMP-1 (Matrix Metalloproteinase-1, up to +309%). In contrast, oligosaccharide fractions had no effect on GAGs synthesis but exhibited similarities for collagens and MMP-1 regulation. At the transcriptomic level, the decrease of COL1A1 and COL1A2 expression, and increase of COL3A1 and MMP-1 expression, confirmed the modulation of ECM metabolism by both fractions. Our research emphasizes that poly- and oligosaccharide Ulva sp. fractions exhibit interesting biological activities and supports their potential use in the area of skin renewal for anti-aging dermo-cosmetic applications.

Staphylococcus epidermidis and Cutibacterium acnes: Two Major Sentinels of Skin Microbiota and the Influence of Cosmetics.

Dermatological and cosmetics fields have recently started to focus on the human skin microbiome and microbiota, since the skin microbiota is involved in the health and dysbiosis of the skin ecosystem. Amongst the skin microorganisms, Staphylococcus epidermidis and Cutibacterium acnes, both commensal bacteria, appear as skin microbiota sentinels. These sentinels have a key role in the skin ecosystem since they protect and prevent microbiota disequilibrium by fighting pathogens and participate in skin homeostasis through the production of beneficial bacterial metabolites. These bacteria adapt to changing skin microenvironments and can shift to being opportunistic pathogens, forming biofilms, and thus are involved in common skin dysbiosis, such as acne or atopic dermatitis. The current evaluation methods for cosmetic active ingredient development are discussed targeting these two sentinels with their assets and limits. After identification of these objectives, research of the active cosmetic ingredients and products that maintain and promote these commensal metabolisms, or reduce their pathogenic forms, are now the new challenges of the skincare industry in correlation with the constant development of adapted evaluation methods.

An Analysis of the Nutritional and Health Values of Caulerpa racemosa (Forsskål) and Ulva fasciata (Delile)-Two Chlorophyta Collected from the Philippines.

Polysaccharides, lipids and amino acid profiles were investigated to understand the nutritional value of Caulerpa racemosa and Ulva fasciata from the Philippines. The results revealed that both species contain high amounts of proteins (8.8-19.9% for C. racemosa and 8.0-11.1% for U. fasciata). The portions of the total amino acids that were essential amino acids (EAAs) (45.28 ± 0.12% for C. racemosa and 42.17 ± 0.12% for U. fasciata) out were comparable to FAO/WHO requirements. Leucine, valine, isoleucine, and lysine are the dominant EAAs in C. racemosa, while leucine, valine, lysine, and phenylalanine are those in U. fasciata. The fatty acid profiles are dominated by monounsaturated fatty acids and polyunsaturated fatty acids in C. racemosa (56.2%), while saturated fatty acids (72.1%) are dominant in U. fasciata. High C18/C20 polyunsaturated fatty acid ratios were recorded in both species. Mineral contents for both seaweeds were within levels considered safe for functional foods. Total pigment content of C. racemosa (140.84 mg/g dw) was almost 20 times higher than that of U. fasciata (7.54 mg/g dw). Hot water extract (HWE) from C. racemosa showed in vitro antiherpetic activity without cytotoxicity. Nutritional characteristics confirmed that C. racemosa could be potentially used as a nutritious and functional food items for human consumption.

Bioactivity of compounds secreted by symbiont bacteria of Nudibranchs from Indonesia.

The aims of this work are to isolate bacterial symbionts from nudibranchs and subsequently to determine anti-Methicillin resistant Staphylococcus aureus (MRSA), cytotoxicity and anti-Herpes simplex virus type 1 (HSV-1) activities of bio compounds. A total of 15 species of nudibranchs were collected from Karimunjawa and five species from Bali, respectively. A total of 245 bacteria isolates were obtained. The anti-MRSA activity screening activity indicated two active bacteria. Ethyl acetate extracts from supernatants, indicating extracelullar compounds, showed an inhibition zone against MRSA at concentrations of 500-1,000 µg/ml. DNA sequence analysis showed that the strain KJB-07 from Phyllidia coelestis was closely related to Pseudoalteromonas rubra, whereas the strain NP31-01 isolated from Phyllidia varicosa was closely related to Virgibacillus salarius. The extract of Pseudoalteromonas rubra was cytotoxic to Vero cells at a concentration of 75 µg/ml. The extract of V. salarius presented no cytotoxicity at concentrations of 5-1,000 µg/ml. No anti HSV-1 was observed for both isolated bacteria. This is the first study describing research on anti-MRSA, cytotoxicity and anti HSV-1 activity of bacterial symbionts from the viscera of nudibranch. Compounds produced by Pseudoalteromonas rubra and V. salarius, had potential anti-MRSA activity. However, only extracts from Pseudoalteromonas rubra showed cytotoxic effects on Vero cells. Three compounds were identified by LC/MS after purification from culture supernatant.

Production of Active Poly- and Oligosaccharidic Fractions from Ulva sp. by Combining Enzyme-Assisted Extraction (EAE) and Depolymerization.

Data on fractionation and depolymerization of the matrix ulvan polysaccharides, and studies on the biological activities on skin cells, are very scarce. In this work, crude ulvans were produced by using EAE (enzyme-assisted extraction) and compared to maceration (an established procedure). After different fractionation procedures-ethanolic precipitation, dialysis, or ammonium sulfate precipitation-the biochemical composition showed that EAE led to an increased content in ulvans. Coupling EAE to sulfate ammonium precipitation led to protein enrichment. Oligosaccharides were obtained by using radical depolymerization by H2O2 and ion-exchange resin depolymerization. Sulfate groups were partially cleaved during these chemical treatments. The potential bioactivity of the fractions was assessed using a lipoxygenase inhibition assay for anti-inflammatory activity and a WST-1 assay for human dermal fibroblast viability and proliferation. All ulvans extracts, poly- and oligosaccharidic fractions from EAE, expanded the fibroblast proliferation rate up to 62%. Our research emphasizes the potential use of poly- and oligosaccharidic fractions of Ulva sp. for further development in cosmetic applications.

Evaluation of immunomodulatory activities of essential oils by high content analysis.

Nowadays, research concerning immunomodulatory products are of great interest, particularly in the treatment of inflammatory diseases or the prevention of infectious diseases. These activities are usually evaluated on cell cultures, by tracking different factors requiring dedicated manipulation. Evaluation of the immunomodulatory activities of essential oils and pure compounds using several technics adapted to high content analysis is described in this study. This approach allows a multiparametric evaluation on a single cell batch, in order to obtain an overall response. The developed method is based on the simultaneous evaluation of phagocytosis, production of iNOS and secretion of IL-6, induced by contact of RAW 264.7 cells with LPS. The results highlight the immunomodulatory activities of cinnamon and clove essential oils. They also provide information, particularly concerning the inhibitory activity of mint essential oil, which inhibits the LPS-induced phagocytosis of RAW 264.7 cells by 42%, at 100 µg/ml. This work presents for the first time the adaptation of high content analyses for the monitoring of immunomodulatory activities of essential oils. This protocol could be adjustable to other cell types and supplemented by the evaluation of additional parameters.

Photo-protective compounds in red macroalgae from Brittany: Considerable diversity in mycosporine-like amino acids (MAAs).

To cope with the biotic and abiotic stresses experienced within their environment, marine macroalgae have developed certain defence mechanisms including the synthesis of photo-protective molecules against light and particularly harmful UV radiation. The aim of this study was to screen selected red algae, a highly diverse phylogenetic group, for the production of photo-protective molecules. The pigment content and composition (i.e. chlorophyll-a, phycobiliproteins and carotenoids) and the composition of mycosporine-like amino acids (MAAs) were studied in 40 species of red macroalgae collected in Brittany (France), at two distinct periods (i.e. February and July 2017). A high inter-specific variability was demonstrated in terms of pigment content and MAA composition. Twenty-three potential MAAs were detected by HPLC, and six were identified by LC-MS (i.e. shinorine, palythine, asterina-330, porphyra-334, usurijene and palythene). This is the first study to report on the composition of pigments and MAAs in a diverse group of red seaweeds from Brittany, including some species for which the MAA composition has never been studied before. Nevertheless, the results suggested that some species of red algae are more likely to cope with high levels of light radiation since those species such as Bostrychia scorpioides, Porphyra dioica, Gracilaria vermiculophylla and Vertebrata lanosa are living in environments exposed to higher levels of irradiation, and had various MAAs in addition to their photo-protective pigments.

Environmentally Friendly Valorization of Solieria filiformis (Gigartinales, Rhodophyta) from IMTA Using a Biorefinery Concept.

Marine macroalgae (seaweed) are an excellent source of novel bioactive metabolites. The biorefinery concept applied to seaweed facilitates the extraction of many chemical constituents from the same biomass ensuring that the resource is used fully, generating few residues through a succession of extraction steps. In the present study, the biomass of the carragenophyte Solieria filiformis (Rhodophyta, Gigartinales) cultured in an integrated multi-trophic aquaculture (IMTA) system was evaluated to obtain valuable products by a biorefinery approach. Enzymatic-assisted extraction (EAE) and microwave-assisted extraction (MAE) were the eco-friendly technologies used to ensure an environmentally friendly valorization of the biomass. Three valuable products were successfully recovered: a water-soluble extract rich in proteins and sulfated polysaccharides suitable as a food supplement; a lipid fraction rich in polyunsaturated fatty acids (PUFAs) with potential to be used in the nutraceutical industry; and a pure ι-carrageenan with a powerful antiviral activity against Herpes simplex virus (EC50 = 6.3 µg mL-1) comparable to the commercial antiviral acyclovir (EC50 = 3.2⁻5.4 µg mL-1).

Total phenolic content and biological activities of enzymatic extracts from Sargassum muticum (Yendo) Fensholt.

Seaweeds are potentially excellent sources of bioactive metabolites that could represent useful leads in the development of new functional ingredients in pharmaceutical and cosmetic industries. In the last decade, new marine bioprocess technologies have allowed the isolation of substances with biological properties. The brown alga Sargassum muticum (Yendo) Fensholt (Ochrophyta) was enzymatically hydrolyzed to prepare water-soluble extracts by using six different commercially available carbohydrate-degrading enzymes and two proteases. Evaluation of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) antioxidant, tyrosinase, elastase, and biofilm inhibition, antibacterial and antiviral activities as well as evaluation of cytotoxicity were realized for each extract. Total phenolic content was measured for extract characterization, and solid-phase extraction was useful to purify the enzymatic extract. Soluble total phenolic content of S. muticum Viscozyme extract was highest with 6.4% of dry weight. Enzymatic Celluclast and Viscozyme extracts had the lowest value of DPPH IC50 indicating a strong antiradical activity, 0.6 mg mL-1, in comparison with other enzymes. The ferric reducing antioxidant power ranged between 48.7 µM Fe2+ Eq, digested with Viscozyme, and 60.8 µM Fe2+ Eq, digested with Amyloglucosidase. Tyrosinase inhibition activity of S. muticum Neutrase extract was 41.3% higher compared to other enzymes. Elastase inhibition activity of S. muticum Shearzyme extract had highest activity (32.8%). All enzymatic extracts showed no cytotoxic effect towards the kidney Vero cells. Meanwhile, only S. muticum Neutrase and Alcalase extracts exhibited potential antiviral activity. In addition, S. muticum Viscozyme and Shearzyme extracts showed promising activity in suppressing the biofilm formation against Pseudomonas aeruginosa and Escherichia coli, respectively. Purification of S. muticum Viscozyme extracts by solid-phase extraction managed to concentrate the phenolic content and improve the bioactivity. These results indicate the promising potential of enzyme-assisted followed by solid-phase extraction in recovering phenolic content and in improving its bioactivity.

Lipid Composition, Fatty Acids and Sterols in the Seaweeds Ulva armoricana, and Solieria chordalis from Brittany (France): An Analysis from Nutritional, Chemotaxonomic, and Antiproliferative Activity Perspectives.

Lipids from the proliferative macroalgae Ulva armoricana (Chlorophyta) and Solieria chordalis (Rhodophyta) from Brittany, France, were investigated. The total content of lipids was 2.6% and 3.0% dry weight for U. armoricana and S. chordalis, respectively. The main fractions of S. chordalis were neutral lipids (37%) and glycolipids (38%), whereas U. armoricana contained mostly neutral lipids (55%). Polyunsaturated fatty acids (PUFA) represented 29% and 15% of the total lipids in U. armoricana and S. chordalis, respectively. In both studied algae, the phospholipids were composed of PUFA for 18%. In addition, PUFA were shown to represent 9% and 4.5% of glycolipids in U. armoricana and S. chordalis, respectively. The essential PUFA were 16:4n-3, 18:4n-3, 18:2n-3, 18:2n-6, and 22:6n-3 in U. armoricana, and 20:4n-6 and 20:5n-3 in S. chordalis. It is important to notice that six 2-hydroxy-, three 3-hydroxy-, and two monounsaturated hydroxy fatty acids were also identified and may provide a chemotaxonomic basis for algae. These seaweeds contained interesting compounds such as squalene, α-tocopherol, cholest-4-en-3-one and phytosterols. The antiproliferative effect was evaluated in vitro on human non-small-cell bronchopulmonary carcinoma line (NSCLC-N6) with an IC50 of 23 µg/mL for monogalactosyldiacylglycerols isolated from S. chordalis and 24 µg/mL for digalactosyldiacylglycerols from U. armoricana. These results confirm the potentialities of valorization of these two species in the fields of health, nutrition and chemotaxonomy.

Lipopolysaccharides from Commensal and Opportunistic Bacteria: Characterization and Response of the Immune System of the Host Sponge Suberites domuncula.

Marine sponges harbor a rich bacterioflora with which they maintain close relationships. However, the way these animals make the distinction between bacteria which are consumed to meet their metabolic needs and opportunistic and commensal bacteria which are hosted is not elucidated. Among the elements participating in this discrimination, bacterial cell wall components such as lipopolysaccharides (LPS) could play a role. In the present study, we investigated the LPS chemical structure of two bacteria associated with the sponge Suberites domuncula: a commensal Endozoicomonas sp. and an opportunistic Pseudoalteromonas sp. Electrophoretic patterns indicated different LPS structures for these bacteria. The immunomodulatory lipid A was isolated after mild acetic acid hydrolysis. The electrospray ionization ion-trap mass spectra revealed monophosphorylated molecules corresponding to tetra- and pentaacylated structures with common structural features between the two strains. Despite peculiar structural characteristics, none of these two LPS influenced the expression of the macrophage-expressed gene S. domuncula unlike the Escherichia coli ones. Further research will have to include a larger number of genes to understand how this animal can distinguish between LPS with resembling structures and discriminate between bacteria associated with it.

Enzyme-assisted extraction of bioactive material from Chondrus crispus and Codium fragile and its effect on herpes simplex virus (HSV-1).

Codium fragile and Chondrus crispus are, respectively, green and red seaweeds which are abundant along the North Atlantic coasts. We investigated the chemical composition and antiviral activity of enzymatic extracts of C. fragile (CF) and C. crispus (CC). On a dry weight basis, CF consisted of 11% protein, 31% neutral sugars, 0.8% sulfate, 0.6% uronic acids, and 49% ash, while CC contained 27% protein, 28% neutral sugars, 17% sulfate, 1.8% uronic acids, and 25% ash. Enzyme-assisted hydrolysis improved the extraction efficiency of bioactive materials. Commercial proteases and carbohydrases significantly improved (p ≤ 0.001) biomass yield (40%-70% dry matter) as compared to aqueous extraction (20%-25% dry matter). Moreover, enzymatic hydrolysis enhanced the recovery of protein, neutral sugars, uronic acids, and sulfates. The enzymatic hydrolysates exhibited significant activity against Herpes simplex virus (HSV-1) with EC50 of 77.6-126.8 µg/mL for CC and 36.5-41.3 µg/mL for CF, at a multiplicity of infection (MOI) of 0.001 ID50/cells without cytotoxity (1-200 µg/mL). The extracts obtained from proteases (P1) and carbohydrases (C3) were also effective at higher virus MOI of 0.01 ID50/cells without cytotoxity. Taken together, these results indicate the potential application of enzymatic hydrolysates of C. fragile and C. crispus in functional food and antiviral drug discovery.

In vitro effects of triclosan and methyl-triclosan on the marine gastropod Haliotis tuberculata.

Triclosan (2,4,4'-trichloro-2'-hydroxy-diphenyl ether; TCS) is an antibacterial agent incorporated in a wide variety of household and personal care products. Because of its partial elimination in sewage treatment plants, TCS is commonly detected in natural waters and sediments. Moreover, due to its high hydrophobicity, TCS accumulates in fatty tissues in various aquatic organisms. TCS can be converted into methyl-triclosan (2,4,4'-trichloro-2'-methoxydiphenyl ether; MTCS) after biological methylation. In this study, the acute cytotoxicity of TCS and MTCS in short-term in vitro experiments was assessed on cell cultures from the European abalone Haliotis tuberculata. The results showed that morphology and density of hemocyte are affected from a concentration of 8 µM TCS. Using the XTT reduction assay, TCS has been demonstrated to decrease hemocyte metabolism activity in a dose- and time-dependent exposure. The IC(50) was evaluated at 6 µM for both hemocyte and gill cells after a 24 h-incubation with TCS. A significant cytotoxicity of MTCS was also observed from 4 µM in 24 h-old hemocyte culture. Our results reveal a toxic effect of TCS and MTCS on immune (hemocytes) and/or respiratory cells (gill cells) of the abalone, species living in coastal waters areas and exposed to anthropogenic pollution.

Occurrence and toxicity of antimicrobial triclosan and by-products in the environment.

INTRODUCTION AND AIMS: A review was undertaken on the occurrence, toxicity, and degradation of triclosan (TCS; 5-chloro-2,4-dichlorophenoxy)phenol) in the environment. TCS is a synthetic, broad-spectrum antibacterial agent incorporated in a wide variety of household and personal care products such as hand soap, toothpaste, and deodorants but also in textile fibers used in a range of other consumer products (e.g., toys, undergarments and cutting boards among other things). OCCURRENCE: Because of its partial elimination in sewage treatment plants, most reports describe TCS as one of the most commonly encountered substances in solid and water environmental compartments. It has been detected in a microgram per liter or microgram per kilogram level in sewage treatment plants (influents, effluents, and sludges), natural waters (rivers, lakes, and estuarine waters), and sediments as well as in drinking water. TOXICITY: Moreover, due to its high hydrophobicity, TCS can accumulate in fatty tissues and has been found in fish and human samples (urine, breast milk, and serum). TCS is known to be biodegradable, photo-unstable, and reactive towards chlorine and ozone. DISCUSSION: As a consequence, it can be transformed into potentially more toxic and persistent compounds, such as chlorinated phenols and biphenyl ethers after chlorination, methyl triclosan after biological methylation, and chlorinated dibenzodioxins after photooxidation. The toxicity of TCS toward aquatic organisms like fish, crustaceans, and algae has been demonstrated with EC50 values near TCS environmental concentrations. It has even been shown to produce cytotoxic, genotoxic, and endocrine disruptor effects. CONCLUSION: Furthermore, the excessive use of TCS is suspected to increase the risk of emergence of TCS-resistant bacteria and the selection of resistant strains.

Lipid A components from Pseudomonas aeruginosa PAO1 (serotype O5) and mutant strains investigated by electrospray ionization ion-trap mass spectrometry.

The lipid A components of the Pseudomonas aeruginosa strains PAO1 (wild-type) and derived mutants PAO1 algC::tet and PAO1 PDO100 were isolated after mild acetic acid hydrolysis of LPS. Their structural heterogeneities were characterized using electrospray ionization (ESI) ion-trap mass spectrometry (MS) with direct infusion in the negative ion mode without prior derivatization. The ESI-mass spectra revealed monophosphorylated molecules corresponding to known tetra-, penta- and hexaacylated structures of P. aeruginosa lipid A. The MS/MS fragmentation patterns allowed the location of fatty acyl chains on the disaccharide backbone of lipid A. In addition, a hexaacylated lipid A containing a hexadecanoyl chain was detected for the first time in strain P. aeruginosa PAO1. With multiple stages of fragmentation (MS(n)), the position of this hexadecanoyl chain O-linked to the decanoyl chain at the C-3(') position of the glucosamine backbone was determined. This sensitive method is suitable to reveal lipid A heterogeneity, i.e. the nature, number and distribution of acyl chains, without prior lipopolysaccharide purification. The lipid A from mutant strains were also characterized and significant differences were shown in the abundance of monophosphorylated lipid A components between the wild-type and the mutant strains.

Structure of a highly phosphorylated lipopolysaccharide core in the Delta algC mutants derived from Pseudomonas aeruginosa wild-type strains PAO1 (serogroup O5) and PAC1R (serogroup O3).

Lipopolysaccharides (LPS) were isolated from rough-type mutant strains of Pseudomonas aeruginosa (Delta algC) derived from wild-type strains PAO1 (serogroup O5) and PAC1R (serogroup O3). Structural studies of the LPS core region with a special focus on the phosphorylation pattern were performed by 2D NMR spectroscopy, including a 1H,(31)P HMQC-TOCSY experiment, MALDI-TOF MS, and Fourier-transform ion cyclotron resonance ESIMS using the capillary skimmer dissociation technique. Both LPS were found to contain two residues each of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) and L-glycero-D-manno-heptose (Hep), one residue of N-(L-alanyl)-D-galactosamine and one O-carbamoyl group (Cm) on the distal Hep residue. The following structures of a tetrasaccharide trisphosphate from strain PAC1R Delta algC and that carrying an additional ethanolamine phosphate group (PEtN) from strain PAO1 Delta algC were elucidated: [carbohydrate structre: see text] where R=P in PAC1R Delta algC and PPEtN in PAO1 Delta algC. To our knowledge, in this work the presence of ethanolamine diphosphate is unambiguously confirmed and its position established for the first time in the LPS core of a rough-type strain of P. aeruginosa. In addition, the structure of the complete LPS core of wild-type strain P. aeruginosa PAO1 was reinvestigated and the position of the phosphorylation sites was revised.