scholarly journals Zinc Import Mediated by AdcABC is Critical for Colonization of the Dental Biofilm by Streptococcus mutans in an Animal Model

2021 ◽  
Author(s):  
Tridib Ganguly ◽  
Alexandra M. Peterson ◽  
Jessica K. Kajfasz ◽  
Jacqueline Abranches ◽  
José A. Lemos
Keyword(s):  
Trace Metals ◽  
Metal Binding ◽  
Pathogenic Bacteria ◽  
Bioinformatic Analysis ◽  
Preliminary Evidence ◽  
Infection Model ◽  
Limiting Factor ◽  
Uptake System ◽  
Dental Biofilm ◽  
High Concentrations

SummaryTrace metals are essential to all domains of life but toxic when found at high concentrations. While the importance of iron in host-pathogen interactions is firmly established, contemporary studies indicate that other trace metals, including manganese and zinc, are also critical to the infectious process. In this study, we sought to identify and characterize the zinc uptake system(s) of S. mutans, a keystone pathogen in dental caries and a causative agent of bacterial endocarditis. Different than other pathogenic bacteria, including several streptococci, that encode multiple zinc import systems, bioinformatic analysis indicated that the S. mutans core genome encodes a single, highly conserved, zinc importer commonly known as AdcABC. Inactivation of the genes coding for the metal-binding AdcA (ΔadcA) or both AdcC ATPase and AdcB permease (ΔadcCB) severely impaired the ability of S. mutans to grow under zinc-depleted conditions. Intracellular metal quantifications revealed that both mutants accumulated less zinc when grown in the presence of a sub-inhibitory concentration of a zinc-specific chelator. Notably, the ΔadcCB strain displayed a severe colonization defect in a rat oral infection model. Both Δadc strains were hypersensitive to high concentrations of manganese, showed reduced peroxide tolerance, and formed less biofilm in sucrose-containing media when cultivated in the presence of the lowest amount of zinc that support their growth, but not when zinc was supplied in excess. Collectively, this study identifies AdcABC as the lone high affinity zinc importer of S. mutans and provides preliminary evidence that zinc is a growth-limiting factor within the dental biofilm.

scholarly journals Characterization of Simultaneous Uptake of Xylose and Glucose in Caldicellulosiruptor Kronotskyensis for Optimal Hydrogen Production

2021 ◽  
Author(s):  
Thitiwut Vongkampang ◽  
Krishnan Sreeni ◽  
Jonathan Engvall ◽  
Carl Grey ◽  
Ed van Niel
Keyword(s):  
Glucose Uptake ◽  
Bioinformatic Analysis ◽  
Growth Patterns ◽  
Sugar Uptake ◽  
Uptake System ◽  
Sugar Mixtures ◽  
Xylose Uptake ◽  
Growth Profiles ◽  
High Concentrations

Abstract BackgroundCaldicellulosiruptor kronotskyensis has gained interest for its ability to grow on various lignocellulosic biomass. The aim of this study was to investigate the growth profiles of C . kronotskyensis in the presence of mixtures of glucose-xylose. Recently, we characterized a diauxic-like pattern for C. saccharolyticus on lignocellulosic sugar mixtures. In this study we aimed to investigated further whether C . kronotskyensis has adapted to uptake glucose in the disaccharide form (cellobiose) rather than the monosaccharide (glucose). ResultsInterestingly, growth of C . kronotskyensis on glucose and xylose mixtures did not display diauxic-like growth patterns. Closer investigation revealed that, in contrast to C. saccharolyticus , C . kronotskyensis does not possess a second uptake system for glucose. Both C. saccharolyticus and C . kronotskyensis share the characteristics of preferring xylose over glucose. Growth on xylose was twice as fast (μ max = 0.57 h -1 ) as on glucose (μ max = 0.28 h -1 ). It was found that C . kronotskyensis takes up glucose and xylose simultaneously with the same transporter. A study of the sugar uptake was made with different glucose-xylose ratios to find a kinetic relationship between the two sugars for transport into the cell. High concentrations of glucose inhibited xylose uptake and vice versa. The inhibition constants were estimated to be K I,glu = 0.01 cmol·L -1 and K I,xyl = 0.001 cmol·L -1 , hence glucose uptake was more severely inhibited by xylose uptake. Bioinformatic analysis indicated the lack of another sugar uptake system in C . kronotskyensis as compared to C. saccharolyticus . Therefore, it was investigated whether glucose uptake by C . kronotskyensis was in the form of cellobiose. Indeed, cellobiose is taken up faster than glucose, nevertheless, the growth rate on each sugar remained similar. ConclusionsC . kronotskyensis possesses a xylose transporter that might take up glucose at an inferior rate even in the absence of xylose. Alternatively, glucose can be taken up in the form of cellobiose, but growth performance is still inferior to growth on xylose. Therefore, we propose that the catabolism of C . kronotskyensis has adapted more strongly to pentose rather than hexose thereby having obtained a specific survival edge in thermophilic lignocellulosic degradation communities.

scholarly journals Microbiome-based environmental monitoring of a dairy processing facility highlights the challenges associated with low microbial-load samples

2021 ◽  
Vol 5 (1) ◽  
Author(s):  
Aoife J. McHugh ◽  
Min Yap ◽  
Fiona Crispie ◽  
Conor Feehily ◽  
Colin Hill ◽  
...  
Keyword(s):  
Environmental Monitoring ◽  
Illumina Sequencing ◽  
Food Chain ◽  
Control Measures ◽  
Initial Assessment ◽  
Limiting Factor ◽  
Accurate Identification ◽  
Processing Facility ◽  
High Concentrations ◽  
Sequencing Platforms

AbstractEfficient and accurate identification of microorganisms throughout the food chain can potentially allow the identification of sources of contamination and the timely implementation of control measures. High throughput DNA sequencing represents a potential means through which microbial monitoring can be enhanced. While Illumina sequencing platforms are most typically used, newer portable platforms, such as the Oxford Nanopore Technologies (ONT) MinION, offer the potential for rapid analysis of food chain microbiomes. Initial assessment of the ability of rapid MinION-based sequencing to identify microbes within a simple mock metagenomic mixture is performed. Subsequently, we compare the performance of both ONT and Illumina sequencing for environmental monitoring of an active food processing facility. Overall, ONT MinION sequencing provides accurate classification to species level, comparable to Illumina-derived outputs. However, while the MinION-based approach provides a means of easy library preparations and portability, the high concentrations of DNA needed is a limiting factor.

scholarly journals Interactions between Rainbow Trout Eyed Eggs and Flavobacterium spp. Using a Bath Challenge Model: Preliminary Evaluation of Bacteriophages as Pathogen Control Agents

2021 ◽  
Vol 9 (5) ◽  
pp. 971
Author(s):  
Valentina L. Donati ◽  
Inger Dalsgaard ◽  
Anniina Runtuvuori-Salmela ◽  
Heidi Kunttu ◽  
Johanna Jørgensen ◽  
...  
Keyword(s):  
Rainbow Trout ◽  
Pathogenic Bacteria ◽  
Infection Model ◽  
Preliminary Evaluation ◽  
Fish Eggs ◽  
Short Term ◽  
Rainbow Trout Oncorhynchus Mykiss ◽  
Pathogen Control ◽  
Number Of Bacteria ◽  
Bath Challenge

The microbial community surrounding fish eyed eggs can harbor pathogenic bacteria. In this study we focused on rainbow trout (Oncorhynchus mykiss) eyed eggs and the potential of bacteriophages against the pathogenic bacteria Flavobacterium psychrophilum and F. columnare. An infection bath method was first established, and the effects of singular phages on fish eggs was assessed (survival of eyed eggs, interaction of phages with eyed eggs). Subsequently, bacteria-challenged eyed eggs were exposed to phages to evaluate their effects in controlling the bacterial population. Culture-based methods were used to enumerate the number of bacteria and/or phages associated with eyed eggs and in the surrounding environment. The results of the study showed that, with our infection model, it was possible to re-isolate F. psychrophilum associated with eyed eggs after the infection procedure, without affecting the survival of the eggs in the short term. However, this was not possible for F. columnare, as this bacterium grows at higher temperatures than the ones recommended for incubation of rainbow trout eyed eggs. Bacteriophages do not appear to negatively affect the survival of rainbow trout eyed eggs and they do not seem to strongly adhere to the surface of eyed eggs either. Finally, the results demonstrated a strong potential for short term (24 h) phage control of F. psychrophilum. However, further studies are needed to explore if phage control can be maintained for a longer period and to further elucidate the mechanisms of interactions between Flavobacteria and their phages in association with fish eggs.

scholarly journals Trk2 Potassium Transport System in Streptococcus mutans and Its Role in Potassium Homeostasis, Biofilm Formation, and Stress Tolerance

2016 ◽  
Vol 198 (7) ◽  
pp. 1087-1100 ◽  
Author(s):  
Gursonika Binepal ◽  
Kamal Gill ◽  
Paula Crowley ◽  
Martha Cordova ◽  
L. Jeannine Brady ◽  
...  
Keyword(s):  
Stress Tolerance ◽  
Streptococcus Mutans ◽  
Transport System ◽  
Biofilm Formation ◽  
Cellular Response ◽  
Pathogenic Bacteria ◽  
Transport Systems ◽  
Content Type ◽  
Growth And Survival ◽  
Dental Biofilm

ABSTRACTPotassium (K+) is the most abundant cation in the fluids of dental biofilm. The biochemical and biophysical functions of K+and a variety of K+transport systems have been studied for most pathogenic bacteria but not for oral pathogens. In this study, we establish the modes of K+acquisition inStreptococcus mutansand the importance of K+homeostasis for its virulence attributes. TheS. mutansgenome harbors four putative K+transport systems that included two Trk-like transporters (designated Trk1 and Trk2), one glutamate/K+cotransporter (GlnQHMP), and a channel-like K+transport system (Kch). Mutants lacking Trk2 had significantly impaired growth, acidogenicity, aciduricity, and biofilm formation. [K+] less than 5 mM eliminated biofilm formation inS. mutans. The functionality of the Trk2 system was confirmed by complementing anEscherichia coliTK2420 mutant strain, which resulted in significant K+accumulation, improved growth, and survival under stress. Taken together, these results suggest that Trk2 is the main facet of the K+-dependent cellular response ofS. mutansto environment stresses.IMPORTANCEBiofilm formation and stress tolerance are important virulence properties of caries-causingStreptococcus mutans. To limit these properties of this bacterium, it is imperative to understand its survival mechanisms. Potassium is the most abundant cation in dental plaque, the natural environment ofS. mutans. K+is known to function in stress tolerance, and bacteria have specialized mechanisms for its uptake. However, there are no reports to identify or characterize specific K+transporters inS. mutans. We identified the most important system for K+homeostasis and its role in the biofilm formation, stress tolerance, and growth. We also show the requirement of environmental K+for the activity of biofilm-forming enzymes, which explains why such high levels of K+would favor biofilm formation.

scholarly journals Characterization of simultaneous uptake of xylose and glucose in Caldicellulosiruptor kronotskyensis for optimal hydrogen production

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Thitiwut Vongkampang ◽  
Krishnan Sreenivas ◽  
Jonathan Engvall ◽  
Carl Grey ◽  
Ed W. J. van Niel
Keyword(s):  
Glucose Uptake ◽  
Growth Patterns ◽  
Sugar Uptake ◽  
Uptake System ◽  
Sugar Mixtures ◽  
Xylose Uptake ◽  
Growth Profiles ◽  
High Concentrations ◽  
Xylose Transporter

Abstract Background Caldicellulosiruptor kronotskyensis has gained interest for its ability to grow on various lignocellulosic biomass. The aim of this study was to investigate the growth profiles of C. kronotskyensis in the presence of mixtures of glucose–xylose. Recently, we characterized a diauxic-like pattern for C. saccharolyticus on lignocellulosic sugar mixtures. In this study, we aimed to investigate further whether C. kronotskyensis has adapted to uptake glucose in the disaccharide form (cellobiose) rather than the monosaccharide (glucose). Results Interestingly, growth of C. kronotskyensis on glucose and xylose mixtures did not display diauxic-like growth patterns. Closer investigation revealed that, in contrast to C. saccharolyticus, C. kronotskyensis does not possess a second uptake system for glucose. Both C. saccharolyticus and C. kronotskyensis share the characteristics of preferring xylose over glucose. Growth on xylose was twice as fast (μmax = 0.57 h−1) as on glucose (μmax = 0.28 h−1). A study of the sugar uptake was made with different glucose–xylose ratios to find a kinetic relationship between the two sugars for transport into the cell. High concentrations of glucose inhibited xylose uptake and vice versa. The inhibition constants were estimated to be KI,glu = 0.01 cmol L−1 and KI,xyl = 0.001 cmol L−1, hence glucose uptake was more severely inhibited by xylose uptake. Bioinformatics analysis could not exclude that C. kronotskyensis possesses more than one transporter for glucose. As a next step it was investigated whether glucose uptake by C. kronotskyensis improved in the form of cellobiose. Indeed, cellobiose is taken up faster than glucose; nevertheless, the growth rate on each sugar remained similar. Conclusions C. kronotskyensis possesses a xylose transporter that might take up glucose at an inferior rate even in the absence of xylose. Alternatively, glucose can be taken up in the form of cellobiose, but growth performance is still inferior to growth on xylose. Therefore, we propose that the catabolism of C. kronotskyensis has adapted more strongly to pentose rather than hexose, thereby having obtained a specific survival edge in thermophilic lignocellulosic degradation communities.

scholarly journals Amycomicin is a potent and specific antibiotic discovered with a targeted interaction screen

2018 ◽  
Vol 115 (40) ◽  
pp. 10124-10129 ◽  
Author(s):  
Gleb Pishchany ◽  
Emily Mevers ◽  
Sula Ndousse-Fetter ◽  
Dennis J. Horvath ◽  
Camila R. Paludo ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Pathogenic Bacteria ◽  
Fatty Acid Biosynthesis ◽  
Streptomyces Coelicolor ◽  
Specific Inhibitor ◽  
Mouse Skin ◽  
Infection Model ◽  
Microbial Species ◽  
Rdna Sequencing ◽  
New Antibiotics

The rapid emergence of antibiotic-resistant pathogenic bacteria has accelerated the search for new antibiotics. Many clinically used antibacterials were discovered through culturing a single microbial species under nutrient-rich conditions, but in the environment, bacteria constantly encounter poor nutrient conditions and interact with neighboring microbial species. In an effort to recapitulate this environment, we generated a nine-strain actinomycete community and used 16S rDNA sequencing to deconvolute the stochastic production of antimicrobial activity that was not observed from any of the axenic cultures. We subsequently simplified the community to just two strains and identifiedAmycolatopsissp. AA4 as the producing strain andStreptomyces coelicolorM145 as an inducing strain. Bioassay-guided isolation identified amycomicin (AMY), a highly modified fatty acid containing an epoxide isonitrile warhead as a potent and specific inhibitor ofStaphylococcus aureus. Amycomicin targets an essential enzyme (FabH) in fatty acid biosynthesis and reducesS. aureusinfection in a mouse skin-infection model. The discovery of AMY demonstrates the utility of screening complex communities against specific targets to discover small-molecule antibiotics.

Ferritin Crystals in the Gut Caeca of a Deep-Sea Hydrothermal Vent Stegocephalid (Crustacea: Amphipoda)

1997 ◽  
Vol 77 (1) ◽  
pp. 269-272 ◽  
Author(s):  
P.G. Moore ◽  
P.S. Rainbow
Keyword(s):  
Trace Metals ◽  
Hydrothermal Vent ◽  
Tectonic Activity ◽  
Amphipod Species ◽  
Single Pair ◽  
Thin Sections ◽  
Widespread Occurrence ◽  
X Ray ◽  
Mid Atlantic Ridge ◽  
High Concentrations

Ferritin crystals and calcium granules are reported from the ventral ceaca of Steleuthera ecoprophycea (Amphipoda: Stegocephalidae) collected from the Snake Pit hydrothermal vent, Mid-Atlantic Ridge (3520 m).In a series of earlier papers (Moore & Rainbow, 1984, 1989, 1992; Moore et al., 1994), the authors described the widespread occurrence of intracellular, octahedral crystals of ferritin in the ventral caeca of a range of stegocephalid amphipod species from the continental shelf epibenthos and oceanic plankton. The discovery at 3500 m of a new Steleuthera species (S. ecoprophycea), from a hydrothermal vent on the Mid-Atlantic Ridge was announced recently by Bellan-Santini & Thurston (1996), and a complete description is provided therein.Oceanic ridge sites are notable for their tectonic activity and the presence of a diversity of trace metals at high concentrations is to be expected in hydrothermal plumes emanating from such regions (German & Angel, 1995). The detoxification of accumulated trace metals in the ventral ceaca of stegocephalid amphipods from uncontaminated environments is now relatively well known (see above), so it was of interest to investigate whether a vent stegocephalid showed an atypical presence of trace metals in detoxified form in cells of the ventral caeca. The results following from an opportunity to investigate this are reported herein.Material was derived from a dive made by the submersible ‘Alvin’ at Snake Pit hydrothermal field, Mid-Atlantic Ridge (23°23′N 44°56′W), 3520 m, on 16 June 1993 (see Bellan-Santini & Thurston, 1996). Steleuthera ecoprophycea was preserved in 70% alcohol. Subsequently to dissection, the single pair of ventral caeca from each of the four damaged amphipods investigated were post-fixed in 4% glutaraldehyde. For electron microscopy and x-ray microanalysis in STEM mode, ventral caecal tissues were then dehydrated through 95% and absolute ethanol, cleared in propylene oxide, embedded in TAAB resin, sectioned at 0·5 μm (semi-thin sections) on a Reichert OmU2 ultramicrotome and examined without staining in a JEOL 100C electron microscope equipped with LINK system EDX energy dispersive x-ray microanalyser.

scholarly journals Characterization of an ATP Translocase Identified in the Destructive Plant Pathogen “Candidatus Liberibacter asiaticus”

2009 ◽  
Vol 192 (3) ◽  
pp. 834-840 ◽  
Author(s):  
Cheryl M. Vahling ◽  
Yongping Duan ◽  
Hong Lin
Keyword(s):  
Plant Pathogen ◽  
Bioinformatic Analysis ◽  
Intracellular Bacteria ◽  
Cell Competition ◽  
Candidatus Liberibacter ◽  
Citrus Huanglongbing ◽  
Liberibacter Asiaticus ◽  
High Concentrations ◽  
Obligate Intracellular Bacteria

ABSTRACT ATP/ADP translocases transport ATP across a lipid bilayer, which is normally impermeable to this molecule due to its size and charge. These transport proteins appear to be unique to mitochondria, plant plastids, and obligate intracellular bacteria. All bacterial ATP/ADP translocases characterized thus far have been found in endosymbionts of protozoa or pathogens of higher-order animals, including humans. A putative ATP/ADP translocase was uncovered during the genomic sequencing of the intracellular plant pathogen “Candidatus Liberibacter asiaticus,” the causal agent of citrus huanglongbing. Bioinformatic analysis of the protein revealed 12 transmembrane helices and predicted an isoelectric point of 9.4, both of which are characteristic of this family of proteins. The “Ca. Liberibacter asiaticus” gene (nttA) encoding the translocase was subsequently expressed in Escherichia coli and shown to enable E. coli to import ATP directly into the cell. Competition assays with the heterologous E. coli system demonstrated that the translocase was highly specific for ATP and ADP but that other nucleotides, if present in high concentrations, could also be taken up and/or block the ability of the translocase to import ATP. In addition, a protein homologous to NttA was identified in “Ca. Liberibacter solanacearum,” the bacterium associated with potato zebra chip disease. This is the first reported characterization of an ATP translocase from “Ca. Liberibacter asiaticus,” indicating that some intracellular bacteria of plants also have the potential to import ATP directly from their environment.

scholarly journals The zinc transporter ZnuABC is critical for the virulence of Chromobacterium violaceum and contributes to diverse zinc-dependent physiological processes

2021 ◽  
Author(s):  
Renato E. R. S. Santos ◽  
Waldir P. da Silva Júnior ◽  
Simone A. Harrison ◽  
Eric P Skaar ◽  
Walter J. Chazin ◽  
...  
Keyword(s):  
Mutant Strain ◽  
Zinc Uptake ◽  
Chromobacterium Violaceum ◽  
Host Protein ◽  
Infection Model ◽  
Uptake System ◽  
Environmental Bacterium ◽  
Mouse Infection Model ◽  
Synthetic Metal

Chromobacterium violaceum is a ubiquitous environmental bacterium that causes sporadic life-threatening infections in humans. How C. violaceum acquires zinc to colonize environmental and host niches is unknown. In this work, we demonstrated that C. violaceum employs the zinc uptake system ZnuABC to overcome zinc limitation in the host, ensuring the zinc supply for several physiological demands. Our data indicated that the C. violaceum ZnuABC transporter is encoded in a zur-CV_RS15045-CV_RS15040-znuCBA operon. This operon was repressed by the zinc uptake regulator Zur and derepressed in the presence of the host protein calprotectin (CP) and the synthetic metal chelator EDTA. A ΔznuCBA mutant strain showed impaired growth under these zinc-chelated conditions. Moreover, the deletion of znuCBA provoked a reduction in violacein production, swimming motility, biofilm formation, and bacterial competition. Remarkably, the ΔznuCBA mutant strain was highly attenuated for virulence in an in vivo mouse infection model and showed a low capacity to colonize the liver, grow in the presence of CP, and resist neutrophil killing. Overall, our findings demonstrate that ZnuABC is essential for C. violaceum virulence, contributing to subvert the zinc-based host nutritional immunity.

scholarly journals Mycobacterial resistance to zinc poisoning requires assembly of P-ATPase-containing membrane metal efflux platforms

2021 ◽  
Author(s):  
Yves-Marie Boudehen ◽  
Marion Faucher ◽  
Xavier Marechal ◽  
Roger Miras ◽  
Jerome Rech ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Metal Binding ◽  
Bacterial Resistance ◽  
Binding Motif ◽  
Membrane Microdomain ◽  
High Zinc ◽  
Functional Membrane ◽  
Zinc Concentrations ◽  
Metal Efflux ◽  
High Concentrations

Transition metals are toxic at high concentrations. The P1B-ATPase metal exporter CtpC/Rv3270 is required for resistance to zinc poisoning in the human pathogen Mycobacterium tuberculosis. Here, we discovered that zinc resistance also depends on the chaperone-like protein PacL1/Rv3269. PacL1 bound Zn2+, but unlike PacL1 and CtpC, the PacL1 metal-binding motif (MBM) was required only at high zinc concentrations. PacL1 co-localized with CtpC in dynamic microdomains within the mycobacterial plasma membrane. Microdomain formation did not require flotillins nor the PacL1 MBM. Instead, loss of the PacL1 Glutamine/Alanine repeats led to loss of CtpC and sensitivity to zinc. PacL1 and CtpC are within the same operon, and homologous PacL1-P1B-ATPase pairs are widely distributed within and across prokaryotes. PacL1 colocalized and functioned redundantly with PacL orthologs in Mycobacterium tuberculosis. Overall, our study suggests that PacL proteins are scaffolds that assemble P-ATPase-containing metal efflux platforms, a novel type of functional membrane microdomain that underlies bacterial resistance to metal poisoning.

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