Pore-forming treatments induce aggregation of Salmonella Senftenberg through protein leakage.

Fresh herbs are not commonly associated with foodborne pathogens, due to the production of essential oils with antimicrobial activity. Recalls of contaminated basil, and basil outbreaks caused by Salmonella motivated studies aimed to comprehend the antimicrobial activity of basil essential oils, and to explore the mechanisms in which Salmonella can overcome them. Linalool, a major constituent of basil oil, increases the permeability of Salmonella Senftenberg cells by damaging their membrane. Linalool also induces bacterial aggregation. We hypothesized that the membrane perforation effect triggers cell aggregation through leakage of intracellular substances from live and dead cells. By exposing S. Senftenberg to additional physical (sonication) or chemical (eugenol, Triton-X-100) treatments, we showed that the aggregation is caused by various membrane-targeted treatments. Enzymatic degradation of leaked proteins restricted the bacterial aggregation, and disassembled existing aggregates. Moreover, supplemented proteins such as bacterial intracellular proteins or BSA also caused aggregation, further supporting the hypothesis that non-specific proteins trigger the bacterial aggregation. This study provides a novel understanding of the role of protein leakage in promoting bacterial aggregation. Since aggregation has significant roles in food safety and microbial ecology, this finding may establish future studies about microbial resistance via formation of clusters similar to biofilm development.

Adaptation of Salmonella enterica Serovar Senftenberg to Linalool and Its Association with Antibiotic Resistance and Environmental Persistence.

A clinical isolate of Salmonella enterica serovar Senftenberg, isolated from an outbreak linked to the herb Ocimum basilicum L. (basil), has been shown to be resistant to basil oil and to the terpene alcohol linalool. To better understand how human pathogens might develop resistance to linalool and to investigate the association of this resistance with resistance to different antimicrobial agents, selective pressure was applied to the wild-type strain by sequential exposure to increasing concentrations of linalool. The results demonstrated that S Senftenberg adapted to linalool with a MIC increment of at least 8-fold, which also resulted in better resistance to basil oil and better survival on harvested basil leaves. Adaptation to linalool was shown to confer cross protection against the antibiotics trimethoprim, sulfamethoxazole, piperacillin, chloramphenicol, and tetracycline, increasing their MICs by 2- to 32-fold. The improved resistance was shown to correlate with multiple phenotypes that included changes in membrane fatty acid composition, induced efflux, reduced influx, controlled motility, and the ability to form larger aggregates in the presence of linalool. The adaptation to linalool obtained in vitro did not affect survival on the basil phyllosphere in planta and even diminished survival in soil, suggesting that development of extreme resistance to linalool may be accompanied by a loss of fitness. Altogether, this report notes the concern regarding the ability of human pathogens to develop resistance to commercial essential oils, a resistance that is also associated with cross-resistance to antibiotics and may endanger public health.IMPORTANCE Greater consumer awareness and concern regarding synthetic chemical additives have led producers to control microbial spoilage and hazards by the use of natural preservatives, such as plant essential oils with antimicrobial activity. This report establishes, however, that these compounds may provoke the emergence of resistant human pathogens. Herein, we demonstrate the acquisition of resistance to basil oil by Salmonella Senftenberg. Exposure to linalool, a component of basil oil, resulted in adaptation to the basil oil mixture, as well as cross protection against several antibiotics and better survival on harvested basil leaves. Collectively, this work highlights the hazard to public health while using plant essential oils without sufficient knowledge about their influence on pathogens at subinhibitory concentrations.

Mechanisms of resistance to linalool in Salmonella Senftenberg and their role in survival on basil.

Fresh produce contaminated with human pathogens raises vital and ecological questions about bacterial survival strategies. Such occurrence was basil harboring Salmonella enterica serovar Senftenberg that caused an outbreak in 2007. This host was unanticipated due to its production of antibacterial substances, including linalool. We show that linalool perforates bacterial membranes, resulting in increased permeability and leakage of vital molecules. It also inhibits cell motility and causes bacterial aggregation. Linalool-resistance was investigated by identification and characterization of S. Senftenberg mutants that perform altered resistance. Resistance mechanisms include selective permeability, regulated efflux/influx and chemotaxis-controlled motility. Moreover, survival of S. Senftenberg on basil leaves was substantially affected by McpL, a putative chemotaxis-related receptor, and RfaG, a component of the lipopolysaccharide production pathway, both have a role in resistance to linalool. Results reveal that adaptation to linalool occurs in nature by concurrent mechanisms. This adaption raises concerns about pathogens adaptation to new hosts including antimicrobial-compound-producing plants.

Resistance to essential oils affects survival of Salmonella enterica serovars in growing and harvested basil.

The number of outbreaks of food-borne illness associated with consumption of fresh products has increased. A recent and noteworthy outbreak occurred in 2007. Basil contaminated with Salmonella enterica serovar Senftenberg was the source of this outbreak. Since basil produces high levels of antibacterial compounds the aim of this study was to investigate if the emerging outbreak reflects ecological changes that occurred as a result of development of resistance to ingredients of the basil oil. We irrigated basil plants with contaminated water containing two Salmonella serovars, Typhimurium and Senftenberg, and showed that Salmonella can survive on the basil plants for at least 100 days. S. Senftenberg counts in the phyllosphere were significantly higher than S. Typhimurium, moreover, S. Senftenberg was able to grow on stored harvested basil leaves. Susceptibility experiments demonstrated that S. Senftenberg is more resistant to basil oil and to its antimicrobial constituents: linalool, estragole and eugenol. This may indicate that S. Senftenberg had adapted to the basil environment by developing resistance to the basil oil. The emergence of resistant pathogens has a significant potential to change the ecology, and opens the way for pathogens to survive in new niches in the environment such as basil and other plants.

Effect of high ß-palmitate content in infant formula on the intestinal microbiota of term infants.

OBJECTIVES: Palmitic acid (PA) constitutes 17% to 25% of the human milk fatty acids, and ~70% is esterified in the sn-2 position of triglycerides (ß-palmitate). In the sn-2 position, PA is not hydrolyzed and thus is efficiently absorbed. The PA in palm oils, commonly used in infant formulas, is esterified in the sn-1 and sn-3 positions. In these positions, PA is hydrolyzed and forms poorly absorbed calcium complexes. The present study assessed whether high ß-palmitate in infant formulas affects the intestinal flora. METHODS: Thirty-six term infants were enrolled: 14 breast-fed (BF group) and 22 formula-fed infants who were randomly assigned to receive formula containing high ß-palmitate (HBP group, n=14), or low ß-palmitate (LBP group, n=8), where 44% and 14% of the PA was ß-palmitate, respectively. The total amount of PA in the formulas was 19% and 22% in the LBP and HBP groups, respectively. Neither formula contained pre- or probiotics. Stool samples were collected at enrollment and at 6 weeks for the quantification of bacteria. RESULTS: At 6 weeks, the HBP and BF groups had higher Lactobacillus and bifidobacteria counts than the LBP group (P<0.01). The Lactobacillus counts at 6 weeks were not significantly different between the HBP and BF groups. Lactobacillus counts were 1.2×10¹°, 1.2×10¹¹, and 5.6×10¹° CFU/g for LBP, HBP, and BF groups, respectively. Bifidobacteria counts were 5.1×109, 1.2×10¹¹, and 3.9×10¹° CFU/g for LBP, HBP, and BF groups, respectively. CONCLUSIONS: HBP formula beneficially affected infant gut microbiota by increasing the Lactobacillus and bifidobacteria counts in fecal stools.

The effects of stainless steel finish on Salmonella Typhimurium attachment, biofilm formation and sensitivity to chlorine.

Bacterial colonization and biofilm formation on stainless steel (SS) surfaces can be sources for cross contamination in food processing facilities, possessing a great threat to public health and food quality. Here the aim was to demonstrate the influence of surface finish of AISI 316 SS on colonization, biofilm formation and susceptibility of Salmonella Typhimurium to disinfection. Initial attachment of S. Typhimurium on surfaces of SS was four times lower, when surface was polished by Bright-Alum (BA) or Electropolishing (EP), as compared to Mechanical Sanded (MS) or the untreated surface (NT). The correlation between roughness and initial bacterial attachment couldn't account on its own to explain differences seen. Biofilms with similar thickness (15-18 µm) were developed on all surfaces 1-day post inoculation, whereas EP was the least covered surface (23%). Following 5-days, biofilm thickness was lowest on EP and MS (30 µm) and highest on NT (62 µm) surfaces. An analysis of surface composition suggested a link between surface chemistry and biofilm development, where the higher concentrations of metal ions in EP and MS surfaces correlated with limited biofilm formation. Interestingly, disinfection of biofilms with chlorine was up to 130 times more effective on the EP surface (0.005% surviving) than on the other surfaces. Overall these results suggest that surface finish should be considered carefully in a food processing plant.

Presence and persistence of Salmonella enterica serotype typhimurium in the phyllosphere and rhizosphere of spray-irrigated parsley.

Salmonella enterica is one of the major food-borne pathogens associated with ready-to-eat fresh foods. Although polluted water might be a significant source of contamination in the field, factors that influence the transfer of Salmonella from water to the crops are not well understood, especially under conditions of low pathogen levels in water. The aim of this study was to investigate the short- and long-term (1 h to 28 days) persistence of Salmonella enterica serotype Typhimurium in the phyllosphere and the rhizosphere of parsley following spray irrigation with contaminated water. Plate counting and quantitative real-time PCR (qRT-PCR)-based methods were implemented for the quantification. By applying qRT-PCR with enrichment, we were able to show that even irrigation with water containing as little as ∼300 CFU/ml resulted in the persistence of S. Typhimurium on the plants for 48 h. Irrigation with water containing 8.5 log CFU/ml resulted in persistence of the bacteria in the phyllosphere and the rhizosphere for at least 4 weeks, but the population steadily declined with a major reduction in bacterial counts, of ∼2 log CFU/g, during the first 2 days. Higher levels of Salmonella were detected in the phyllosphere when plants were irrigated during the night compared to irrigation during the morning and during winter compared to the other seasons. Further elucidation of the mechanisms underlying the transfer of Salmonella from contaminated water to crops, as well as its persistence over time, will enable the implementation of effective irrigation and control strategies.

Binding Capabilities of Different Genetically Engineered pVIII Proteins of the Filamentous M13/Fd Virus and Single-Walled Carbon Nanotubes.

Binding functional biomolecules to non-biological materials, such as single-walled carbon nanotubes (SWNTs), is a challenging task with relevance for different applications. However, no one has yet undertaken a comparison of the binding of SWNTs to different recombinant filamentous viruses (phages) bioengineered to contain different binding peptides fused to the virus coat proteins. This is important due to the range of possible binding efficiencies and scenarios that may arise when the protein's amino acid sequence is modified, since the peptides may alter the virus's biological properties or they may behave differently when they are in the context of being displayed on the virus coat protein; in addition, non-engineered viruses may non-specifically adsorb to SWNTs. To test these possibilities, we used four recombinant phage templates and the wild type. In the first circumstance, we observed different binding capabilities and biological functional alterations; e.g., some peptides, in the context of viral templates, did not bind to SWNTs, although it was proven that the bare peptide did. The second circumstance was excluded, as the wild-type virus was found to hardly bind to the SWNTs. These results may be relevant to the possible use of the virus as a "SWNT shuttle" in nano-scale self-assembly, particularly since the pIII proteins are free to act as binding-directing agents. Therefore, knowledge of the differences between and efficiencies of SWNT binding templates may help in choosing better binding phages or peptides for possible future applications and industrial mass production.

Factors affecting the thermal resistance of Salmonella Typhimurium in tahini.

Salmonella enterica is a leading human pathogen responsible for foodborne outbreaks worldwide. In the last decade, foods with low water activity (aw) and high-fat content have been involved in an increased occurrence of foodborne outbreaks. This research focuses on the foodstuff tahini, which is often linked to Salmonella infection outbreaks and recalls. Thermal treatments are suggested to reduce microbial populations in tahini, but little is known about its effectiveness against Salmonella. Our major objectives were to study the survival of Salmonella Typhimurium in tahini treated at temperatures ≥ 70 °C, and to identify food related factors that could influence its survival. Based on our experimental results the thermal treatments at 70 °C, 80 °C and 90 °C are suitable to inactivate only a partial population of Salmonella. The death of Salmonella in tahini matches a biphasic logarithmic inactivation model, with a maximal 3-log reduction after 1 h at 90 °C. Moreover, we observed that a second thermal treatment the day after the first treatment, is significantly less effective compared with the first thermal treatment. The inactivation rates of Salmonella in 100% tahini are almost 4-log lower than in water/tahini emulsions at 70 °C, with negative linear correlation between D-value and aw, and the Salmonella susceptibility to heat in sesame oil/tahini emulsions is affected by the matrix of pre-acclimation. Bacteria that had been acclimated in tahini kept their heat resistance, while acclimation in sesame oil before mixing in the preheated oil/tahini emulsions resulted in a sharp decline within 2 min at 70 °C. According to these findings, tahini producers' current pasteurization processes are not sufficient to achieve the required 5-log reduction. Furthermore, we suggest that due to the tahini heterogenicity, the aw in the micro-environment of each bacterium, which is shaped by the tahini substances, plays an essential role in Salmonella's survival in tahini at temperatures ≥ 70 °C.

The activity of BcsZ of Salmonella Typhimurium and its role in Salmonella-plants interactions.

Salmonella enterica is one of the most common human pathogens associated with fresh produce outbreaks. The present study suggests that expression of BcsZ, one of the proteins in the bcs complex, enhances the survival of Salmonella Typhimurium on parsley. BcsZ demonstrated glucanase activity with the substrates carboxymethylcellulose and crystalline cellulose, and was responsible for a major part of the S. Typhimurium CMCase activity. Moreover, there was constitutive expression of BcsZ, which was also manifested after exposure to plant polysaccharides and parsley-leaf extract. In an in-planta model, overexpression of BcsZ significantly improved the epiphytic and endophytic survival of S. Typhimurium on/in parsley leaves compared with the wild-type strain and bcsZ null mutant. Interestingly, necrotic lesions appeared on the parsley leaf after infiltration of Salmonella overexpressing BcsZ, while infiltration of the wild-type S. Typhimurium did not cause any visible symptoms. Infiltration of purified BcsZ enzyme, or its degradation products also caused symptoms on parsley leaves. We suggest that the BcsZ degradation products trigger the plant's defense response, causing local necrotic symptoms. These results indicate that BcsZ plays an important role in the Salmonella-plant interactions, and imply that injured bacteria may take part in these interactions.

Burkholderiaceae and Multidrug Resistance Genes Are Key Players in Resistome Development in a Germfree Soil Model.

Assembly of a resistome in parallel with the establishment of a microbial community is not well understood. Germfree models can reveal microbiota interactions and shed light on bacterial colonization and resistance development under antibiotic pressure. In this study, we exposed germfree soil (GS), GS with diluted nontreated soil (DS), and nontreated soil (NS) to various concentrations of tetracycline (TET) in a nongermfree environment for 10 weeks, followed by 2 weeks of exposure to water. High-throughput sequencing was used to profile bacterial communities and antibiotic resistance genes (ARGs) in the soils. The initial bacterial loads were found to shape the profiles of bacterial communities and the resistomes. GS and DS treated with TET and the same soils left untreated had similar profiles, whereas NS showed different profiles. Soils with the same initial bacterial loads had their profiles shifted by TET treatment. Multidrug resistance (MDR) genes were the most abundant ARG types in all soils, with multidrug efflux pump genes being the discriminatory ARGs in GS regardless of different TET treatments and in GS, DS, and NS after TET. Furthermore, MDR genes were significantly enriched by TET treatment. In contrast, tetracycline resistance genes were either absent or low in relative abundance. The family Burkholderiaceae was predominant in all soils (except in NS treated with water) and was positively selected for by TET treatment. Most importantly, Burkholderiaceae were the primary carrier of ARGs, including MDR genes. IMPORTANCE This is the first study to examine how resistomes develop and evolve using GS. GS can be used to study the colonization and establishment of bacterial communities under antibiotic selection. Surprisingly, MDR genes were the main ARGs detected in GS, and TET treatments did not positively select for specific tetracycline resistance genes. Additionally, Burkholderiaceae were the key bacterial hosts for MDR genes in the current GS model under the conditions investigated. These results show that the family Burkholderiaceae underpins the development of resistome and serves as a source of ARGs. The ease of establishment of Burkholderiaceae and MDR genes in soils has serious implications for human health, since these bacteria are versatile and ubiquitous in the environment.

Salicylate reduces the antimicrobial activity of ciprofloxacin against extracellular Salmonella enterica serovar Typhimurium, but not against Salmonella in macrophages.

OBJECTIVES: Salicylate, a potent inducer of the MarA activator in Salmonella enterica, is the principal metabolite of aspirin, which is often consumed for medicinal and cosmetic uses. Our research was aimed at testing if salicylate activates the mar regulon in macrophage-associated Salmonella (intracellular bacteria), and investigating its effects on bacterial susceptibility to ciprofloxacin extracellularly and intracellularly. METHODS: J774 macrophages were infected with S. enterica serovar Typhimurium (wild-type and marA null mutant), treated with ciprofloxacin with and without pre-exposure to salicylate, and the surviving bacteria were counted. Similar experiments were conducted with bacteria in broth (extracellular bacteria). Phe-Arg-beta-naphthylamide (PAbetaN) was added to investigate the role of efflux pumps in resistance. The transcriptional regulation of marRAB, acrAB and micF in extracellular and intracellular Salmonella Typhimurium with and without salicylate and ciprofloxacin was investigated using green fluorescent protein as a marker protein and quantitative real time PCR. RESULTS: Pre-exposure of Salmonella to salicylate increased the resistance of extracellular but not intracellular bacteria to ciprofloxacin, although salicylate stimulated the expression of mar genes in intracellular and extracellular bacteria. Using marA mutants and the inhibitor PAbetaN, we showed that the improved resistance in extracellular bacteria is derived from the induction of acrAB by salicylate, which is mediated by MarA. CONCLUSIONS: In intracellular bacteria, the expression of acrAB is already higher when compared with extracellular cells; therefore, salicylate does not result in significant acrAB induction intracellularly and subsequent resistance enhancement. Results show that conclusions raised from extracellular studies cannot be applied to intracellular bacteria, although the systems have similar functions.

Cellodextrin and laminaribiose ABC transporters in Clostridium thermocellum.

Clostridium thermocellum is an anaerobic thermophilic bacterium that grows efficiently on cellulosic biomass. This bacterium produces and secretes a highly active multienzyme complex, the cellulosome, that mediates the cell attachment to and hydrolysis of the crystalline cellulosic substrate. C. thermocellum can efficiently utilize only beta-1,3 and beta-1,4 glucans and prefers long cellodextrins. Since the bacterium can also produce ethanol, it is considered an attractive candidate for a consolidated fermentation process in which cellulose hydrolysis and ethanol fermentation occur in a single process. In this study, we have identified and characterized five sugar ABC transporter systems in C. thermocellum. The putative transporters were identified by sequence homology of the putative solute-binding lipoprotein to known sugar-binding proteins. Each of these systems is transcribed from a gene cluster, which includes an extracellular solute-binding protein, one or two integral membrane proteins, and, in most cases, an ATP-binding protein. The genes of the five solute-binding proteins were cloned, fused to His tags, overexpressed, and purified, and their abilities to interact with different sugars was examined by isothermal titration calorimetry. Three of the sugar-binding lipoproteins (CbpB to -D) interacted with different lengths of cellodextrins (G(2) to G(5)), with disassociation constants in the micromolar range. One protein, CbpA, binds only cellotriose (G(3)), while another protein, Lbp (laminaribiose-binding protein) interacts with laminaribiose. The sugar specificity of the different binding lipoproteins is consistent with the observed substrate preference of C. thermocellum, in which cellodextrins (G(3) to G(5)) are assimilated faster than cellobiose.

CsrA and CsrB are required for the post-transcriptional control of the virulence-associated effector protein AvrA of Salmonella enterica.

The virulence-associated effector protein AvrA of Salmonella enterica is an ubiquitin-like acetyltransferase/cysteine protease, which interferes with the first line of immune response of the target organism. In contrast to translation of the AvrA protein in S. enterica strains, which takes place either constitutively (class 1 strains), or after acid induction (class 2 strains), or not at all (class 3 strains); the constitutive transcription of the respective avrA genes occurs regardless of these defined expression classes. When the number of avrA genes and mRNA molecules is raised experimentally using plasmids carrying the respective cloned avrA genes together with their promoter regions, the translation of avrA mRNA takes place very strongly in all respective AvrA expression classes. This kind of copy-dependent, post-transcriptional control of AvrA was shown to be dependent on the regulatory action of the CsrA/CsrB system since the deletion of both genes completely abolished the translation in the tested S. enterica strains, whereas the transcription remained unaffected. Moreover, AvrA production in strains carrying the cloned avrA genes on plasmids remained dependent on the presence of CsrA but unaffected in csrB mutant strains. On the other hand, overproduction of the regulatory molecules CsrA and CsrB in S. enterica strains carrying cloned csrA and csrB genes on plasmids ceased the expression of AvrA again. Therefore, the expression of avrA is suggested to be regulated in a post-transcriptional manner by critical and effective concentrations of CsrA (see-saw regulation), which is achieved through the sequestering activity of CsrB.

Transfer of Salmonella enterica serovar Typhimurium from contaminated irrigation water to parsley is dependent on curli and cellulose, the biofilm matrix components.

Enteric pathogens can contaminate fresh produce, and this contaminated produce can be a significant potential source of human illness. The objective of this study was to determine a possible mode of transfer of Salmonella Typhimurium from contaminated irrigation water to mature parsley plants and to investigate the role of bacterial cellulose and curli. Parsley plants were drip irrigated with water containing green fluorescent protein-labeled Salmonella Typhimurium. Stems and leaves were harvested 1 day after the third irrigation and examined for the presence of Salmonella Typhimurium. Three weeks after harvesting, the presence of Salmonella was again confirmed in the regrown plants. During this period, bacterial numbers on leaves declined from 4.1 (+/- 0.3) to 2.3 (+/- 0.1) log CFU g(-1) (P < 0.05). Numbers in the soil were constant (5 log CFU g(-1)). Results demonstrated the ability of Salmonella Typhimurium to transfer from irrigation water to the edible parts of the plants. Confocal laser scanning microscopic images revealed that Salmonella Typhimurium formed aggregates at a depth of 8 to 32 microm beneath the leaf surface. Penetration might be achieved through the roots or the phyllosphere. The importance of the bacterial cellulose and curli was determined by comparing the wild-type strain with its mutants, which lack the ability to synthesize cellulose and curli. Counts of the double mutant were 2-log higher in the soil but 1-log lower in the leaves (P < 0.05). Deletion of the agfBA gene (for curli) was more effective than deletion of bcsA (for cellulose). Thus, curli and cellulose play a role in the transfer or survival of Salmonella Typhimurium in the plant, as they do for plant pathogens.

A comparative study assaying commonly used sanitizers for antimicrobial activity against indicator bacteria and a Salmonella Typhimurium strain on fresh produce.

With increased concerns over failures in vegetable and fruit sanitation, evaluating the efficacy of widely approved chemicals is ever more important. The purpose of this study was to determine whether sanitation treatments are equally effective against indicator bacteria and human enteric pathogens on cucumber and parsley. We provide here an experimental overview on the efficacy of common sanitation methods, which are based on peracetic acid-hydrogen peroxide, sodium dichloroisocyanurate, and the quaternary ammonium compound didecyldimethylammonium chloride. The sanitizers were tested for their activity against natural populations of total aerobic microorganisms, enterococci, and coliforms, and against the enteric pathogen Salmonella Typhimurium ATCC 14028 (which was added artificially). Results revealed that compared with washing parsley and cucumbers with water, treatments with all three sanitizers were not effective, resulting in a maximal reduction of only 0.7 log CFU of Salmonella Typhimurium. These sanitizers were also not effective in removal of natural bacteria from parsley (maximal reduction was 0.7 log CFU). Sanitation of cucumber was more successful; peracetic acid showed the most effective result, with a reduction of 2.7 log in aerobic microorganisms compared with cucumbers washed with water. Still, removal of natural bacteria from cucumbers proved more efficient than the removal of Salmonella Typhimurium. This may create a debate about the necessity of the sanitation and its contribution to safety, because sanitation of some contaminated vegetables may result in an increased likelihood of foods that, although they are given good hygienic ratings due to low microbial counts, harbor pathogens.

Structure-toxicity relationship of aminoglycosides: correlation of 2'-amine basicity with acute toxicity in pseudo-disaccharide scaffolds.

A new pseudo-disaccharide NB23 with a 3',4'-methylidene protection was designed and its properties were evaluated in comparison to other two structurally related pseudo-disaccharides. The basicity of the 2'-amine was found to be well correlated to acute toxicity data in mice: the increase in the basicity is associated with the toxicity increase. Based on these data, a new pseudo-trisaccharide NB45 was constructed. NB45 exhibited significant antibacterial activity while at the same time retained low acute toxicity.

Food restriction followed by refeeding with a casein- or whey-based diet differentially affects the gut microbiota of pre-pubertal male rats.

Researchers are gaining an increasing understanding of host-gut microbiota interactions, but studies of the role of gut microbiota in linear growth are scarce. The aim of this study was to investigate the effect of food restriction and refeeding with different diets on gut microbiota composition in fast-growing rats. Young male Sprague-Dawley rats were fed regular rat chow ad libitum (control group) or subjected to 40% food restriction for 36 days followed by continued restriction or ad libitum refeeding for 24 days. Three different diets were used for refeeding: regular vegetarian protein chow or chow in which the sole source of protein was casein or whey. In the control group, the composition of the microbiota remained stable. Food restriction for 60 days led to a significant change in the gut microbiota at the phylum level, with a reduction in the abundance of Firmicutes and an increase in Bacteroidetes and Proteobacteria. Rats refed with the vegetarian protein diet had a different microbiota composition than rats refed the casein- or whey-based diet. Similarities in the bacterial population were found between rats refed vegetarian protein or a whey-based diet and control rats, and between rats refed a casein-based diet and rats on continued restriction. There was a significant strong correlation between the gut microbiota and growth parameters: humerus length, epiphyseal growth plate height, and levels of insulin-like growth factor 1 and leptin. In conclusion, the type of protein in the diet significantly affects the gut microbiota and, thereby, may affect animal's health.

Effect of triclosan on Salmonella typhimurium at different growth stages and in biofilms.

Triclosan is a potent biocide that is included in a diverse range of products. This research was aimed to investigate the susceptibility of planktonic and biofilm-associated Salmonella enterica serovar Typhimurium to triclosan, and to identify potential mechanisms of adaptation. The effect of triclosan was studied on planktonic Salmonella (log and stationary phases), on biofilm-associated cells, on bacteria derived from disrupted biofilms and on a biofilm-deficient mutant. An eight-log reduction of exponentially growing cells was observed with 1000 micro g mL(-1) triclosan within 10 min, a 3.6-log reduction in stationary cells and a 6.3-log reduction in stationary cells of a biofilm-deficient mutant (P<0.05). Biofilm-associated cells were tolerant (1-log reduction). However, biofilm-derived cells showed sensitivity to triclosan similar to stationary-phase cells. Triclosan induced the transcription of fabI and micF. Within biofilms, triclosan also up-regulated the transcription of acrAB, encoding for an efflux pump, marA, and the cellulose-synthesis-coding genes bcsA and bcsE. Thus, Salmonella within biofilms could experience reduced influx, increased efflux and enhanced exopolysaccharides production. Our results demonstrated that the tolerance of Salmonella towards triclosan in the biofilm was attributed to low diffusion through the extracellular matrix, while changes of gene expression might provide further resistance to triclosan and to other antimicrobials.

Draft Genome Sequence of Salmonella enterica Serovar Senftenberg 070885 and Its Linalool-Adapted Mutant.

Here we report the genome sequences of both Salmonella Senftenberg 070885, a clinical isolate from the 2007 outbreak linked to basil, and its mutant linalool-adapted S Senftenberg (LASS). These draft genomes of S Senftenberg may enable the identification of bacterial genes responsible for resistance to basil oil.