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occurrence of vibrio parahaemolyticus, vibrio cholerae and vibrio vulnificus in the clam ruditapes philippinarum (adams & reeve, 1850) from emilia romagna and sardinia, italy.marine vibrios, vibrio parahaemolyticus, v. vulnificus and v. cholerae are responsible of the majority of food-borne human infections by consumption of bivalve shellfish. the aim of the present study was to ascertain the occurrence of these bacteria, and their potential pathogenicity, in the manila clam r. philippinarum from emilia romagna (er) and sardinia (sr) regions, italy. isolation was performed on chromagar(tm) vibrio with subculture on (thiosulfate-citrate-bile salts-sucrose) agar and m- ...201627800436
anoctamin 6 contributes to cl- secretion in accessory cholera enterotoxin (ace)-stimulated diarrhea: an essential role for phosphatidylinositol 4,5-bisphosphate (pip2) signaling in cholera.accessory cholera enterotoxin (ace) of vibrio cholerae has been shown to contribute to diarrhea. however, the signaling mechanism and specific type of cl(-) channel activated by ace are still unknown. we have shown here that the recombinant ace protein induced icl of apical plasma membrane, which was inhibited by classical cacc blockers. surprisingly, an ace-elicited rise of current was neither affected by ano1 (tmem16a)-specific inhibitor t16a(inh)-ao1(tao1) nor by the cystic fibrosis transmemb ...201627799301
a plant from the altiplano of northern chile senecio nutans, inhibits the vibrio cholerae pathogen.in the altiplano of northern chile the plant senecio nutans is habitually used as an infusion to relieve the effects of altitude sickness (locally known as "puna"). it is also used to alleviate the bronchitis, whooping cough, asthma, stomachache, tiredness and fever. the extreme conditions under which these plant grow and scientific data that shows the inhibiting potential of the essential oils of plants of the genus senecio represents great potential in the study of their application to control ...201627795930
nanocarriers from gras zein proteins to encapsulate hydrophobic actives.one factor limiting the expansion of nanomedicines has been the high cost of the materials and processes required for their production. we present a continuous, scalable, low cost nanoencapsulation process, flash nanoprecipitation (fnp) that enables the production of nanocarriers (ncs) with a narrow size distribution using zein corn proteins. zein is a low cost, gras protein (having the fda status of "generally regarded as safe") currently used in food applications, which acts as an effective en ...201627744703
the role of vibrio cholerae haemagglutinin protease (hap) in extra-intestinal infection.based on the diversity of surface o antigen vibrio cholerae can be classified into 206 serogroups. vibrio cholerae is the causative agent of cholera and extra intestinal infections like, septicemia, wound infection and haemorrhagic reactions. pathogenic factors of v. cholerae extra-intestinal infection are yet to be explored.201627790431
cholera cases cluster in time and space in matlab, bangladesh: implications for targeted preventive interventions.cholera remains a serious public health threat in asia, africa and in parts of the americas. three world health organization (who) pre-qualified oral cholera vaccines are now available but their supply is limited, so current supplies must be administered strategically. this requires an improved understanding of disease transmission and control strategies.201627789673
complete genome sequences of two marine vibrio cholerae strains isolated from the south coast of sweden.vibrio cholerae serogroups o1 and o139 are commonly associated with diarrhea, while non-o1-o139 strains may cause wound infections. here, we present the genome sequences of two v. cholerae strains isolated from blue mussels (mytilus edulis) collected in coastal waters of southern sweden.201627789632
adherence to intestinal cells promotes biofilm formation in vibrio cholerae.vibrio cholerae, the etiological agent of cholera, is known to form biofilms to persist in the environment. it is demonstrated here that even during infection, biofilm genes are upregulated, and microscopic observation indicated that biofilm formation is initiated almost immediately after adherence of v. cholerae to intestinal cells. about 7-fold upregulation of the biofilm regulatory gene vpst was observed within 30 minutes of adherence of v. cholerae to the intestinal cell line int 407, and a ...201627638940
vibrio cholerae laboratory infection of the adult house fly musca domestica.the present study was designed to test the hypothesis that house flies may be capable of specifically harbouring ingested vibrio cholerae in their digestive tracts. flies were continuously fed green fluorescent protein (gfp)-labelled, non-o1/non-o139 environmental strains of v. cholerae. bacterial burdens were quantitatively measured using plate counts and localization was directly observed using confocal microscopy. vibrio cholerae were present in the fly alimentary canal after just 4 h, and re ...201627444689
non-toxigenic environmental vibrio cholerae o1 strain from haiti provides evidence of pre-pandemic cholera in hispaniola.vibrio cholerae is ubiquitous in aquatic environments, with environmental toxigenic v. cholerae o1 strains serving as a source for recurrent cholera epidemics and pandemic disease. however, a number of questions remain about long-term survival and evolution of v. cholerae strains within these aquatic environmental reservoirs. through monitoring of the haitian aquatic environment following the 2010 cholera epidemic, we isolated two novel non-toxigenic (ctxa/b-negative) vibrio cholerae o1. these t ...201627786291
a genomic island in vibrio cholerae with vpi-1 site-specific recombination characteristics contains crispr-cas and type vi secretion modules.cholera is a devastating diarrhoeal disease caused by certain strains of serogroup o1/o139 vibrio cholerae. mobile genetic elements such as genomic islands (gis) have been pivotal in the evolution of o1/o139 v. cholerae. perhaps the most important gi involved in cholera disease is the v. cholerae pathogenicity island 1 (vpi-1). this gi contains the toxin-coregulated pilus (tcp) gene cluster that is necessary for colonization of the human intestine as well as being the receptor for infection by t ...201627845364
origins of the current seventh cholera pandemic.vibrio cholerae has caused seven cholera pandemics since 1817, imposing terror on much of the world, but bacterial strains are currently only available for the sixth and seventh pandemics. the el tor biotype seventh pandemic began in 1961 in indonesia, but did not originate directly from the classical biotype sixth-pandemic strain. previous studies focused mainly on the spread of the seventh pandemic after 1970. here, we analyze in unprecedented detail the origin, evolution, and transition to pa ...201627849586
phenotypic and antibiogram pattern of v. cholerae isolates from a tertiary care hospital in mumbai during 2004-2013: a retrospective cross-sectional study.cholera is a major gastroenteric disease with reports on fluctuation and resistance. hence, the objective is to determine the trend in seasonality, resistance pattern, prevalent biotypes, serotypes and phage types between 2004 and 2013 among vibrio cholerae isolates.201627888174
whole-genome sequences of 26 vibrio cholerae isolates.the human pathogen vibrio cholerae employs several adaptive mechanisms for environmental persistence, including natural transformation and type vi secretion, creating a reservoir for the spread of disease. here, we report whole-genome sequences of 26 diverse v. cholerae isolates, significantly increasing the sequence diversity of publicly available v. cholerae genomes.201628007852
effects of chromosomal deletion of the operon encoding the multiple resistance and ph-related antiporter in vibrio cholerae.to examine the possible physiological significance of mrp, a multi-subunit cation/proton antiporter from vibrio cholerae, a chromosomal deletion δmrp of v. cholerae was constructed and characterized. the resulting mutant showed a consistent early growth defect in lb broth that became more evident at elevated ph of the growth medium and increasing na+ or k+ loads. after 24 h incubation, these differences disappeared likely due to the concerted effort of other cation pumps in the mrp mutant. pheno ...201627902431
selective and efficient elimination of vibrio cholerae with a chemical modulator that targets glucose metabolism.vibrio cholerae, a gram-negative bacterium, is the causative agent of pandemic cholera. previous studies have shown that the survival of the seventh pandemic el tor biotype v. cholerae strain n16961 requires production of acetoin in a glucose-rich environment. the production of acetoin, a neutral fermentation end-product, allows v. cholerae to metabolize glucose without a ph drop, which is mediated by the production of organic acid. this finding suggests that inhibition of acetoin fermentation c ...201627900286
quorum regulated resistance of vibrio cholerae against environmental bacteriophages.predation by bacteriophages can significantly influence the population structure of bacterial communities. vibrio cholerae the causative agent of cholera epidemics interacts with numerous phages in the aquatic ecosystem, and in the intestine of cholera patients. seasonal epidemics of cholera reportedly collapse due to predation of the pathogen by phages. however, it is not clear how sufficient number of the bacteria survive to seed the environment in the subsequent epidemic season. we found that ...201627892495
haitian variant tcpa in vibrio cholerae o1 el tor strains in national capital region (india). 201627934812
comparison of antibody immobilization strategies in detection of vibrio cholerae by surface plasmon resonance.surface plasmon resonance immunosensor for the detection of bacterial cells was first reported in 1998 with high detection limit as much as 10(7 )cfu/ml. since then, many efforts have been made aiming to lower the detection limit and improve the sensitivity of detection. the aim of this study was to compare the effect of four most frequently used immobilization strategies, including direct physical adsorption (physisorption), covalent immobilization via self-assembled monolayer (sam) formation, ...201627923270
cross-protection against vibrio cholerae infection by monoclonal antibodies against vibrio vulnificus rtxa1/martxvv.gram-negative vibrio species secrete multifunctional autoprocessing repeats-in-toxin (martx) toxins associated with bacterial pathogenesis. here, the cross-reactivity and cross-protectivity of mabs against v. vulnificus rtxa1/martxvv was evaluated. passive administration of any of these mabs (21ra, 24ra, 46ra, 47ra and 50ra) provided strong protection against lethal v. cholerae infection. interestingly, 24ra and 46ra, which map to the cysteine protease domain of v. cholerae martxvc , inhibited c ...201627921342
nitrosative stress response in vibrio cholerae: role of s-nitrosoglutathione reductase.vibrio cholerae, the causative agent of cholera, poses serious threats to humans worldwide. v. cholerae faces host inflammatory response and encounters nitrosative stress before establishing successful colonization. it is not clear how v. cholerae combats nitric oxide and reactive nitrogen species. in the present study, we used three clinical strains of v. cholerae and tested their nitrosative stress response pattern towards sodium nitroprusside (snp) and s-nitrosoglutathione (gsno). among them, ...201628000045
cholera rapid test with enrichment step has diagnostic performance equivalent to culture.cholera rapid diagnostic tests (rdt) could play a central role in outbreak detection and surveillance in low-resource settings, but their modest performance has hindered their broad adoption. the addition of an enrichment step may improve test specificity. we describe the results of a prospective diagnostic evaluation of the crystal vc rdt (span diagnostics, india) with enrichment step and of culture, each compared to polymerase chain reaction (pcr), during a cholera outbreak in south sudan. rdt ...201627992488
origins of pandemic vibrio cholerae from environmental gene pools.some microorganisms can transition from an environmental lifestyle to a pathogenic one(1-3). this ecological switch typically occurs through the acquisition of horizontally acquired virulence genes(4,5). however, the genomic features that must be present in a population before the acquisition of virulence genes and emergence of pathogenic clones remain unknown. we hypothesized that virulence adaptive polymorphisms (vaps) circulate in environmental populations and are required for this transition ...201627991885
ribavirin suppresses bacterial virulence by targeting lysr-type transcriptional regulators.targeting bacterial virulence mechanisms without compromising bacterial growth is a promising strategy to prevent drug resistance. lysr-type transcriptional regulators (lttrs) possess structural conservation across bacterial species and regulate virulence in numerous pathogens, making them attractive targets for antimicrobial agents. we targeted aphb, a vibrio cholerae lttr, which regulates the expression of genes encoding cholera toxin and toxin-co-regulated pilus for inhibitor designing. since ...201627991578
polymer-based oral rehydration solution for treating acute watery diarrhoea.acute diarrhoea is one of the main causes of morbidity and mortality among children in low-income countries. glucose-based oral rehydration solution (ors) helps replace fluid and prevent further dehydration from acute diarrhoea. since 2004, the world health organization (who) has recommended the osmolarity of less than 270 mosm/l (ors ≤ 270) versus greater than 310 mosm/l formulation (ors ≥ 310). polymer-based ors (for example, prepared using rice or wheat) slowly releases glucose and may be sup ...201627959472
world health organization estimates of the relative contributions of food to the burden of disease due to selected foodborne hazards: a structured expert elicitation.the foodborne disease burden epidemiology reference group (ferg) was established in 2007 by the world health organization (who) to estimate the global burden of foodborne diseases (fbds). this estimation is complicated because most of the hazards causing fbd are not transmitted solely by food; most have several potential exposure routes consisting of transmission from animals, by humans, and via environmental routes including water. this paper describes an expert elicitation study conducted by t ...201626784029
molecular detection of the three major pathogenic vibrio species from seafood products and sediments in tunisia using real-time pcr.vibrio spp. have emerged as a serious threat to human health worldwide. v. parahaemolyticus , v. cholerae , and v. vulnificus pose a considerable public health risk in tunisia because they cause sporadic and epidemic foodborne infections associated with the consumption of raw or undercooked contaminated seafood. more recently, toxr-positive v. alginolyticus was also reported to be a potential source of contaminated seafood. a total of 247 samples, including 113 fishes ( labrus viridis , penaeus ...201628221954
the replication initiator of the cholera pathogen's second chromosome shows structural similarity to plasmid initiators.the conserved dnaa-oric system is used to initiate replication of primary chromosomes throughout the bacterial kingdom; however, bacteria with multipartite genomes evolved distinct systems to initiate replication of secondary chromosomes. in the cholera pathogen, vibrio cholerae, and in related species, secondary chromosome replication requires the rctb initiator protein. here, we show that rctb consists of four domains. the structure of its central two domains resembles that of several plasmid ...201628031373
role of non-coding regulatory rna in the virulence of human pathogenic vibrios.in recent decades, the identification of small non-coding rnas in bacteria has revealed an important regulatory mechanism of gene expression involved in the response to environmental signals and to the control of virulence. in the family vibrionaceae, which includes several human and animal pathogens, small non-coding rnas (srnas) are closely related to important processes including metabolism, quorum sensing, virulence, and fitness. studies conducted in silico and experiments using microarrays ...201628123382
the epidemiology and antimicrobial resistance of cholera cases in iran during 2013.cholera is an endemic diarrheal disease in iran, caused by vibrio cholerae. the epidemiology, transmission route, environmental determinants and antimicrobial resistant pattern of cholera have been changed during recent years. in this study the epidemiology and antimicrobial resistance of cholera in iran during 2013 outbreak was investigated.201628210461
saharan dust nutrients promote vibrio bloom formation in marine surface waters.vibrio is a ubiquitous genus of marine bacteria, typically comprising a small fraction of the total microbial community in surface waters, but capable of becoming a dominant taxon in response to poorly characterized factors. iron (fe), often restricted by limited bioavailability and low external supply, is an essential micronutrient that can limit vibrio growth. vibrio species have robust metabolic capabilities and an array of fe-acquisition mechanisms, and are able to respond rapidly to nutrien ...201627162369
anticancer and antibacterial secondary metabolites from the endophytic fungus penicillium sp. cam64 against multi-drug resistant gram-negative bacteria.the emergence of multiple-drug resistance bacteria has become a major threat and thus calls for an urgent need to search for new effective and safe anti-bacterial agents.201627917206
antibacterial and cytotoxic cytochalasins from the endophytic fungus phomopsis sp. harbored in garcinia kola (heckel) nut.the continuous emergence of multidrug-resistant (mdr) bacteria drastically reduced the efficacy of our antibiotic armory and consequently, increased the frequency of therapeutic failure. the search for bioactive constituents from endophytic fungi against mdr bacteria became a necessity for alternative and promising strategies, and for the development of novel therapeutic solutions. we report here the isolation and structure elucidation of antibacterial and cytotoxic compounds from phomopsis sp., ...201627842536
development of lipopolysaccharide-mimicking peptides and their immunoprotectivity against vibrio cholerae serogroup o1.vibrio cholerae serogroup o1 is the main causative agent of cholera diseases defined by life threatening rice watery diarrhea. cholera routine vaccination has failed in controlling epidemics in developing countries because of their hard and expensive production. in this study, our aim was to investigate phage displayed mimotopes that could mimic v. cholerae lipopolysaccharide (lps). although lps of vibrio, as an endotoxin, can stimulate the immune system, thereby making it a suitable candidate f ...201627766741
local and global consequences of flow on bacterial quorum sensing.bacteria use a chemical communication process called quorum sensing (qs) to control collective behaviours such as pathogenesis and biofilm formation(1,2). qs relies on the production, release and group-wide detection of signal molecules called autoinducers. to date, studies of bacterial pathogenesis in well-mixed cultures have revealed virulence factors and the regulatory circuits controlling them, including the overarching role of qs(3). although flow is ubiquitous to nearly all living systems( ...201627571752
synthesis and biological evaluation of 5,7-dihydroxyflavanone derivatives as antimicrobial agents.a series of 5,7-dihydroxyflavanone derivatives were efficiently synthesized. their antimicrobial efficacy on gram-negative, gram-positive bacteria and yeast were evaluated. among these compounds, most of the halogenated derivatives exhibited the best antimicrobial activity against gram-positive bacteria, the yeast saccharomyces cerevisiae, and the gram-negative bacterium vibrio cholerae. the cytotoxicities of these compounds were low as evaluated on hepg2 cells using a cell viability assay. this ...201627210435
[n-acetyl-β-d-glucosaminidase of vibrio cholerae].study n-acetyl-β-d-glucosaminidase (chitobiase) (ec 3.2.1.30) in strains of vibrio cholerae of o1/non-o1 serogroups of various origin, that is a component of chitinolytic complex taking into account object of isolation and epidemiologic significance of strains.201627228670
antibacterial activity of a novel fatty acid (14e, 18e, 22e, 26e)-methyl nonacosa-14, 18, 22, 26 tetraenoate isolated from amaranthus spinosus.amaranthus spinosus linn. (amaranthaceae), commonly known as ''spiny pigweed'', is used in both indian traditional system and folk medicine for treatment of infectious diseases for a long time in several traditional herbal medicinal preparations. a novel fatty acid [(14e, 18e, 22e, 26e)-methyl nonacosa-14, 18, 22, 26 tetraenoate] is the major metabolite present.201627043360
characterization of mdaφ, a temperate filamentous bacteriophage of neisseria meningitidis.the mechanism by which neisseria meningitidis becomes invasive is not well understood. comparative genomics identified the presence of an 8 kb island in strains belonging to invasive clonal complexes. this island was designated mda for meningococcal disease associated. mda is highly conserved among meningococcal isolates and its analysis revealed a genomic organization similar to that of a filamentous prophage such as ctxφ of vibrio cholerae. subsequent molecular investigations showed that the m ...201626602366
antibacterial and antiviral effectiveness of two household water treatment devices that use monobrominated hydantoinylated polystyrene.many different household water treatment (hwt) methods have been researched and promoted to mitigate the serious burden of diarrheal disease in developing countries. however, hwt methods using bromine have not been extensively evaluated. two gravity-fed hwt devices (aquasure™ and waterbird™) were used to test the antimicrobial effectiveness of halopure(®) br beads (monobrominated hydantoinylated polystyrene) that deliver bromine. as water flows over the beads, reactive bromine species are eluted ...201627959873
isolation, identification, and evaluation of novel probiotic strains isolated from feces of breast-fed infants.to isolate, identify, and evaluate the probiotic properties of lactic acid bacteria (lab) isolated from the feces of breast-fed infants.201626817236
the α-hydroxyketone lai-1 regulates motility, lqs-dependent phosphorylation signalling and gene expression of legionella pneumophila.the causative agent of legionnaires' disease, legionella pneumophila, employs the autoinducer compound lai-1 (3-hydroxypentadecane-4-one) for cell-cell communication. lai-1 is produced and detected by the lqs (legionella quorum sensing) system, comprising the autoinducer synthase lqsa, the sensor kinases lqss and lqst, as well as the response regulator lqsr. lqs-regulated processes include pathogen-host interactions, production of extracellular filaments and natural competence for dna uptake. he ...201626538361
antibacterial activity of aquatic gliding bacteria.the study aimed to screen and isolate strains of freshwater aquatic gliding bacteria, and to investigate their antibacterial activity against seven common pathogenic bacteria. submerged specimens were collected and isolated for aquatic gliding bacteria using four different isolation media (dw, ma, sap2, and vy/2). gliding bacteria identification was performed by 16s rrna gene sequencing and phylogenetic analysis. crude extracts were obtained by methanol extraction. antibacterial activity against ...201626885469
biofilm formation and detachment in gram-negative pathogens is modulated by select bile acids.biofilms are a ubiquitous feature of microbial community structure in both natural and host environments; they enhance transmission and infectivity of pathogens and provide protection from human defense mechanisms and antibiotics. however, few natural products are known that impact biofilm formation or persistence for either environmental or pathogenic bacteria. using the combination of a novel natural products library from the fish microbiome and an image-based screen for biofilm inhibition, we ...201626992172
vibrio cholerae hemagglutinin(ha)/protease: an extracellular metalloprotease with multiple pathogenic activities.vibrio cholerae of serogroup o1 and o139, the etiological agent of the diarrheal disease cholera, expresses the extracellular zn-dependent metalloprotease hemagglutinin (ha)/protease also reported as vibriolysin. this enzyme is also produced by non-o1/o139 (non-cholera) strains that cause mild, sporadic illness (i.e. gastroenteritis, wound or ear infections). orthologs of ha/protease are present in other members of the vibrionaceae family pathogenic to humans and fish. ha/protease belongs to the ...201626952544
species-dependent hydrodynamics of flagellum-tethered bacteria in early biofilm development.monotrichous bacteria on surfaces exhibit complex spinning movements. such spinning motility is often a part of the surface detachment launch sequence of these cells. to understand the impact of spinning motility on bacterial surface interactions, we develop a hydrodynamic model of a surface-bound bacterium, which reproduces behaviours that we observe in pseudomonas aeruginosa, shewanella oneidensis and vibrio cholerae, and provides a detailed dictionary for connecting observed spinning behaviou ...201626864892
genetic control of bacterial biofilms.nearly all bacterial species, including pathogens, have the ability to form biofilms. biofilms are defined as structured ecosystems in which microbes are attached to surfaces and embedded in a matrix composed of polysaccharides, edna, and proteins, and their development is a multistep process. bacterial biofilms constitute a large medical problem due to their extremely high resistance to various types of therapeutics, including conventional antibiotics. several environmental and genetic signals ...201626294280
pasteurella multocida toxin: targeting mast cell secretory granules during kiss-and-run secretion.pasteurella multocida toxin (pmt), a virulence factor of the pathogenic gram-negative bacterium p. multocida, is a 146 kda protein belonging to the a-b class of toxins. once inside a target cell, the a domain deamidates the α-subunit of heterotrimeric g-proteins, thereby activating downstream signaling cascades. however, little is known about how pmt selects and enters its cellular targets. we therefore studied pmt binding and uptake in porcine cultured intestinal mucosal explants to identify su ...201626763205
the unusual redox properties of c-type oxidases.cytochrome cbb3 (also known as c-type) oxidases belong to the family of heme-copper terminal oxidases which couple at the end of the respiratory chain the reduction of molecular oxygen into water and the pumping of protons across the membrane. they are expressed most often at low pressure of o2 and they exhibit a low homology of sequence with the cytochrome aa3 (a-type) oxidases found in mitochondria. their binuclear active site comprises a high-spin heme b3 associated with a cub center. the pro ...201627664317
characterization of prohibitin 1 as a host partner of vibrio vulnificus rtxa1 toxin.rtxa1 toxin, which results in cytoskeletal rearrangement, contact cytotoxicity, hemolysis, tissue invasion, and lethality in mice, is the most potent cytotoxic virulence factor of vibrio vulnificus. bioinformatics analysis of rtxa1 predicted 4 functional domains that presumably performed discrete functions during host cell killing. v. vulnificus rtxa1 has a unique domain designated as rtxa1-d2, corresponding to amino acids 1951-2574, which is absent in vibrio cholerae multifunctional-autoprocess ...201626136468
low prevalence of vibrio cholerae o1 versus moderate prevalence of intestinal parasites in food-handlers working with health care personnel in haiti.food-handlers with poor personal hygiene working in food-service establishments could be potential sources of infection due to pathogenic organisms. in may 2011, a cross-sectional study was undertaken to determine the prevalence of bacteria and intestinal parasites among food-handlers working with cuban health personnel in haiti. stool specimens were collected from 56 food-handlers and samples were examined using standard procedures. of the food handlers, 26.8% had one bacterial or intestinal pa ...201627077312
"non-toxic" proteins of the botulinum toxin complex exert in-vivo toxicity.the botulinum neurotoxin (bont) causes muscle paralysis and is the most potent toxin in nature. bont is associated with a complex of auxiliary "non-toxic" proteins, which constitute a large-sized toxin complex (l-tc). however, here we report that the "non-toxic" complex of serotype d botulinum l-tc, when administered to rats, exerts in-vivo toxicity on small-intestinal villi. moreover, serotype c and d of the "non-toxic" complex, but not bont, induced vacuole-formation in a rat intestinal epithe ...201627507612
clinical features of human immunodeficiency virus-infected patients presenting with cholera in port-au-prince, haiti.human immunodeficiency virus (hiv) infection has been postulated to alter the natural history of cholera, including increased susceptibility to infection, severity of illness, and chronic carriage of vibrio cholerae haiti has a generalized hiv epidemic with an adult hiv prevalence of 1.9% and recently suffered a cholera epidemic. we conducted a prospective study at the cholera treatment center (ctc) of gheskio in haiti to characterize the coinfection. adults admitted at the ctc for acute diarrhe ...201627549637
mucosal vaccination for prevention of hiv infection and aids.most of hiv infections occur via the genital tract or the rectum and hiv replicates at high levels in lymphoid organs and intestinal mucosa, likely requiring a more diversified immunity than pathogens restricted to a single mucosal site, such as the gastrointestinal tract for vibrio cholera, or the respiratory airways for the influenza virus.201626957199
biochemical characterization of an α1,2-colitosyltransferase from escherichia coli o55:h7.colitose, also known as 3,6-dideoxy-l-galactose or 3-deoxy-l-fucose, is one of only five naturally occurring 3,6-dideoxyhexoses. colitose was found in lipopolysaccharide of a number of infectious bacteria, including escherichia coli o55 & o111 and vibrio cholera o22 & o139. to date, no colitosyltransferase (colt) has been characterized, probably due to the inaccessibility of the sugar donor, gdp-colitose. in this study, starting with chemically prepared colitose, 94.6 mg of gdp-colitose was prep ...201626703456
immunogenic properties of trivalent recombinant protein composed of b-subunits of lt, stx-2, and ct toxins.infectious diarrhoea remains an emerging problem in the world health program. among diarrheagenic agents, vibrio cholerae and enterotoxigenic and enterohemorrhagic escherichia coli are critical enteropathogens. ab5 toxin produced by these bacteria, heat-labile enterotoxin (lt), cholera enterotoxin (ct), and shiga-like cytotoxin (stx) can target the immune system and are subunit vaccine candidates. a chemically-synthesized chimeric construct composed of the binding subunits of these toxins (ltb, ...201626970204
prospective study of pathogens in asymptomatic travellers and those with diarrhoea: aetiological agents revisited.travellers' diarrhoea (td) remains the most frequent health problem encountered by visitors to the (sub)tropics. traditional stool culture identifies the pathogen in only 15% of cases. exploiting pcr-based methods, we investigated td pathogens with a focus on asymptomatic travellers and severity of symptoms. pre- and post-travel stools of 382 travellers with no history of antibiotic use during travel were analysed with a multiplex quantitative pcr for salmonella, yersinia, campylobacter, shigell ...201626970046
diversity of clinical and environmental isolates of vibrio cholerae in natural transformation and contact-dependent bacterial killing indicative of type vi secretion system activity.the bacterial pathogen vibrio cholerae can occupy both the human gut and aquatic reservoirs, where it may colonize chitinous surfaces that induce the expression of factors for three phenotypes: chitin utilization, dna uptake by natural transformation, and contact-dependent bacterial killing via a type vi secretion system (t6ss). in this study, we surveyed a diverse set of 53 isolates from different geographic locales collected over the past century from human clinical and environmental specimens ...201626944842
transcriptional repression of the vc2105 protein by vibrio fadr suggests that it is a new auxiliary member of the fad regulon.recently, our group along with others reported that the vibrio fadr regulatory protein is unusual in that, unlike the prototypical fadr product of escherichia coli, which has only one ligand-binding site, vibrio fadr has two ligand-binding sites and represents a new mechanism for fatty acid sensing. the promoter region of the vc2105 gene, encoding a putative thioesterase, was mapped, and a putative fadr-binding site (aa ctg gta aga gca ctt) was proposed. different versions of the fadr regulatory ...201626944841
cholera toxin b subunit pentamer reassembled from escherichia coli inclusion bodies for use in vaccination.the cholera toxin b subunit (ctb) is secreted in its pentameric form from escherichia coli if its leader peptide is replaced with one of e. coli origin. however, the secretion of the pentamer is generally severely impaired when the molecule is mutated or fused to a foreign peptide. therefore, we attempted to regenerate pentameric ctb from the inclusion bodies (ibs) of e. coli. stepwise dialysis of the ibs solubilized in guanidine hydrochloride predominantly generated soluble high-molecular-mass ...201626828455
characterization of vcc-1, a novel ambler class a carbapenemase from vibrio cholerae isolated from imported retail shrimp sold in canada.one of the core goals of the canadian integrated program for antimicrobial resistance surveillance (cipars) is to monitor major meat commodities for antimicrobial resistance. targeted studies with methodologies based on core surveillance protocols are used to examine other foods, e.g., seafood, for antimicrobial resistance to detect resistances of concern to public health. here we report the discovery of a novel ambler class a carbapenemase that was identified in a nontoxigenic strain of vibrio ...201626824956
cytoplasmic heme-binding protein (hutx) from vibrio cholerae is an intracellular heme transport protein for the heme-degrading enzyme, hutz.hutz is a cytoplasmic heme-binding protein from vibrio cholerae. although we have previously identified hutz as a heme-degrading enzyme [uchida, t., et al. (2012) chem. commun. 48, 6741-6743], the heme transport protein for hutz remained unknown. to identify the heme transport protein for hutz, we focused on the heme utilization operon, hutwxz. to this end, we constructed an expression system for hutx in escherichia coli and purified it to homogeneity. an absorption spectral analysis demonstrate ...201626807477
antibiotic use, its resistance in nepal and recommendations for action: a situation analysis.antibiotics are crucial, life-saving medicines in the fight against infectious disease, but resistance to these drugs is growing all over. this article presents key findings from a detailed situation analysis produced by the global antibiotic resistance partnership (garp)-nepal working group. in the absence of nationally-representative surveillance, it is not possible to fully describe antibiotic resistance in the country, but many important bacterial pathogens are highly resistant to most first ...201626744193
structural characterization and biomedical properties of sulfated polysaccharide from the gladius of sepioteuthis lessoniana (lesson, 1831).sulfated polysaccharide was extracted from the internal shell (gladius) of sepioteuthis lessoniana. the sulfated polysaccharide contained 61.3% of carbohydrate, 0.8% of protein, 28.2% of ash and 1.33% of moisture respectively. the elemental composition was analyzed using chns/o analyzer. the molecular weight of sulfated polysaccharide determined through page was found to be as 66 kda. monosaccharides analysis revealed that sulfated polysaccharide was composed of rhamnose, galactose, xylose and g ...201626724686
evaluation of antibacterial activity of plant mediated cao nanoparticles using cissus quadrangularis extract.an assessment of antibacterial activity of greenly synthesized nanoparticles using aqueous stem extract of cissus quadrangularis was carried out. the synthesized nanoparticles were characterized by uv-vis spectroscopy, sem, xrd, ftir and further subjected for antibacterial activity against the pathogens staphylococcus aureus, escherichia coli, klebsiella pneumoniae, pseudomonas aeruginosa, salmonella typhi, shigella dysenteriae and vibrio cholerae. the sem photograph represents cubic and hexagon ...201626723000
in sickness and in health: the relationships between bacteria and bile in the human gut.colonization of a human host with a commensal microbiota has a complex interaction in which bacterial communities provide numerous health benefits to the host. an equilibrium between host and microbiota is kept in check with the help of biliary secretions by the host. bile, composed primarily of bile salts, promotes digestion. it also provides a barrier between host and bacteria. after bile salts are synthesized in the liver, they are stored in the gallbladder to be released after food intake. t ...201627565580
biosensing vibrio cholerae with genetically engineered escherichia coli.cholera is a potentially mortal, infectious disease caused by vibrio cholerae bacterium. current treatment methods of cholera still have limitations. beneficial microbes that could sense and kill the v. cholerae could offer potential alternative to preventing and treating cholera. however, such v. cholerae targeting microbe is still not available. this microbe requires a sensing system to be able to detect the presence of v. cholera bacterium. to this end, we designed and created a synthetic gen ...201627529184
identification and characterization of a putative manganese export protein in vibrio cholerae.manganese plays an important role in the cellular physiology and metabolism of bacterial species, including the human pathogen vibrio cholerae the intracellular level of manganese ions is controlled through coordinated regulation of the import and export of this element. we have identified a putative manganese exporter (vc0022), named mnea (manganese exporter a), which is highly conserved among vibrio spp. an mnea mutant exhibited sensitivity to manganese but not to other cations. under high-man ...201627481926
screening for transmembrane association in divisome proteins using toxgreen, a high-throughput variant of the toxcat assay.toxcat is a widely used genetic assay to study interactions of transmembrane helices within the inner membrane of the bacterium escherichia coli. toxcat is based on a fusion construct that links a transmembrane domain of interest with a cytoplasmic dna-binding domain from the vibrio cholerae toxr protein. interaction driven by the transmembrane domain results in dimerization of the toxr domain, which, in turn, activates the expression of the reporter gene chloramphenicol acetyl transferase (cat) ...201627453198
lcp crystallization and x-ray diffraction analysis of vcmn, a mate transporter from vibrio cholerae.multidrug and toxic compound extrusion (mate) transporters, one of the multidrug exporter families, efflux xenobiotics towards the extracellular side of the membrane. since mate transporters expressed in bacterial pathogens contribute to multidrug resistance, they are important therapeutic targets. here, a mate-transporter homologue from vibrio cholerae, vcmn, was overexpressed in escherichia coli, purified and crystallized in lipidic cubic phase (lcp). x-ray diffraction data were collected to 2 ...201627380372
quantitative microbial risk assessment (qmra) shows increased public health risk associated with exposure to river water under conditions of riverbed sediment resuspension.although higher microbial concentrations have been reported in sediments than in the overlying water column, most quantitative microbial risk assessment (qmra) studies have not clearly indicated the contribution of sediment-borne pathogens to estimated risks. thus, the present study aimed at determining the public health risk associated with exposure to pathogenic bacteria in polluted river water under undisturbed conditions and conditions of sediment resuspension in the apies river, gauteng, so ...201627297265
human fecal and pathogen exposure pathways in rural indian villages and the effect of increased latrine coverage.efforts to eradicate open defecation and improve sanitation access are unlikely to achieve health benefits unless interventions reduce microbial exposures. this study assessed human fecal contamination and pathogen exposures in rural india, and the effect of increased sanitation coverage on contamination and exposure rates. in a cross-sectional study of 60 villages of a cluster-randomized controlled sanitation trial in odisha, india, human and domestic animal fecal contamination was measured in ...201627192358
a single gene of a commensal microbe affects host susceptibility to enteric infection.indigenous microbes inside the host intestine maintain a complex self-regulating community. the mechanisms by which gut microbes interact with intestinal pathogens remain largely unknown. here we identify a commensal escherichia coli strain whose expansion predisposes mice to infection by vibrio cholerae, a human pathogen. we refer to this strain as 'atypical' e. coli (atec) because of its inability to ferment lactose. the atec strain is resistant to reactive oxygen species (ros) and proliferate ...201627173141
strategies to block bacterial pathogenesis by interference with motility and chemotaxis.infections by motile, pathogenic bacteria, such as campylobacter species, clostridium species, escherichia coli, helicobacter pylori, listeria monocytogenes, neisseria gonorrhoeae, pseudomonas aeruginosa, salmonella species, vibrio cholerae, and yersinia species, represent a severe economic and health problem worldwide. of special importance in this context is the increasing emergence and spread of multidrug-resistant bacteria. due to the shortage of effective antibiotics for the treatment of in ...201627000091
an expeditious synthesis of blood-group antigens, abo histo-blood group type ii antigens and xenoantigen oligosaccharides with amino type spacer-arms.blood group oligosaccharides are one of the most clinically important antigen families and they may also act as secondary ligands for bacterial toxins from escherichia coli and vibrio cholerae. herein we report the synthesis of spacered (sp = ch2ch2ch2nh2) glycosides of a antigen {α-d-galnac-(l→3)-[α-l-fuc-(l→2)]-β-d-gal-}, b antigen{α-d-gal-(l→3)-[α-l-fuc-(l→2)]-β-d-gal-}, lewisx{α-d-gal-(l→4)-[α-l-fuc-(l→3)]-β-d-glcnac-}, a type-ii {α-d-galnac-(l→3)-[α-l-fuc-(l→2)]-β-d-gal-(1→4)-β-d-glcnac-}, ...201626572140
regulation of biofilm formation and cellular buoyancy through modulating intracellular cyclic di-gmp levels in engineered cyanobacteria.the second messenger cyclic dimeric (3'→5') gmp (cyclic di-gmp or c-di-gmp) has been implicated in the transition between motile and sessile lifestyles in bacteria. in this study, we demonstrate that biofilm formation, cellular aggregation or flocculation, and cellular buoyancy are under the control of c-di-gmp in synechocystis sp. pcc 6803 (synechocystis) and fremyella diplosiphon. synechocystis is a unicellular cyanobacterium and displays lower levels of c-di-gmp; f. diplosiphon is filamentous ...201626192200
biochemical and structural characterization of the novel sialic acid-binding site of escherichia coli heat-labile enterotoxin lt-iib.the structurally related ab5-type heat-labile enterotoxins of escherichia coli and vibrio cholerae are classified into two major types. the type i group includes cholera toxin (ct) and e. coli lt-i, whereas the type ii subfamily comprises lt-iia, lt-iib and lt-iic. the carbohydrate-binding specificities of lt-iia, lt-iib and lt-iic are distinctive from those of cholera toxin and e. coli lt-i. whereas ct and lt-i bind primarily to the gm1 ganglioside, lt-iia binds to gangliosides gd1a, gd1b and g ...201627562297
competitive survival of escherichia coli, vibrio cholerae, salmonella typhimurium and shigella dysenteriae in riverbed sediments.studies on the survival of bacterial enteric pathogens in riverbed sediments have mostly focused on individual organisms. reports on the competitive survival of these pathogens in riverbed sediments under the same experimental setup are limited. we investigated the survival of escherichia coli, salmonella enterica ser. typhimurium, vibrio cholerae and shigella dysenteriae in riverbed sediments of the apies river. experiments were performed in flow chambers containing three sediment types and con ...201627193001
killing of vibrio cholerae and escherichia coli strains carrying d-mannose-sensitive ligands by mytilus hemocytes is promoted by a multifunctional hemolymph serum protein.in aquatic environments, bivalve mollusks represent an important ecological niche for microorganisms. persistence of bacteria in bivalve tissues partly depends on their capacity to survive the bactericidal activity of the hemolymph due to both cellular (hemocyes) and soluble serum factors (e.g., enzymes, lectins, opsonins). the extrapallial protein (ep) present in serum of mytilus galloprovincialis (mgep) has been recently shown to work as an opsonin promoting d-mannose sensitive (ms) interactio ...201627041371
dna replication in engineered escherichia coli genomes with extra replication origins.the standard outline of bacterial genomes is a single circular chromosome with a single replication origin. from the bioengineering perspective, it appears attractive to extend this basic setup. bacteria with split chromosomes or multiple replication origins have been successfully constructed in the last few years. the characteristics of these engineered strains will largely depend on the respective dna replication patterns. however, the dna replication has not been investigated systematically i ...201627268399
antibacterial activity of polyphenolic fraction of kombucha against enteric bacterial pathogens.the emergence of multi-drug-resistant enteric pathogens has prompted the scientist community to explore the therapeutic potentials of traditional foods and beverages. the present study was undertaken to investigate the efficacy of kombucha, a fermented beverage of sugared black tea, against enterotoxigenic escherichia coli, vibrio cholerae, shigella flexneri and salmonella typhimurium followed by the identification of the antibacterial components present in kombucha. the antibacterial activity w ...201627638313
an unusual cation-binding site and distinct domain-domain interactions distinguish class ii enolpyruvylshikimate-3-phosphate synthases.enolpyruvylshikimate-3-phosphate synthase (epsps) catalyzes a critical step in the biosynthesis of a number of aromatic metabolites. an essential prokaryotic enzyme and the molecular target of the herbicide glyphosate, epspss are the subject of both pharmaceutical and commercial interest. two epsps classes that exhibit low sequence homology, differing substrate/glyphosate affinities, and distinct cation activation properties have previously been described. here, we report structural studies of t ...201626813771
optimization and characterization of the synthetic secondary chromosome synvicii in escherichia coli.learning by building is one of the core ideas of synthetic biology research. consequently, building synthetic chromosomes is the way to fully understand chromosome characteristics. the last years have seen exciting synthetic chromosome studies. we had previously introduced the synthetic secondary chromosome synvicii in escherichia coli. it is based on the replication mechanism of the secondary chromosome in vibrio cholerae. here, we present a detailed analysis of its genetic characteristics and ...201628066763
evolution and structural dynamics of bacterial glycan binding adhesins.infectious disease processes like bacterial adherence or the activity of secreted toxins frequently gain host and tissue specificity by glycan binding interactions with the host glycome. recent functional and structural studies highlight the high niche specialization of bacterial lectins, but also reveal a remarkable plasticity in their glycan binding sites and mechanisms, to adapt to host glycome dynamics or changing environmental conditions at the site of infection. in this review we put empha ...201628043017
single-cell analysis of the plasmablast response to vibrio cholerae demonstrates expansion of cross-reactive memory b cells.we characterized the acute b cell response in adults with cholera by analyzing the repertoire, specificity, and functional characteristics of 138 monoclonal antibodies (mabs) generated from single-cell-sorted plasmablasts. we found that the cholera-induced responses were characterized by high levels of somatic hypermutation and large clonal expansions. a majority of the expansions targeted cholera toxin (ct) or lipopolysaccharide (lps). using a novel proteomics approach, we were able to identify ...201627999163
microbiology, infection control and infection related outcome in pediatric patients in an oncology center in eastern india: experience from tata medical center, kolkata.infection is a major determinant in the outcome of patients with cancer.201626842142
expression and assembly of cholera toxin b subunit and domain iii of dengue virus 2 envelope fusion protein in transgenic potatoes.the rates of mosquito-transmitted dengue virus infection in humans have increased in tropical and sub-tropical areas. domain iii of dengue envelope protein (ediii) is involved in cellular receptor binding and induces serotype-specific neutralizing antibodies. ediii fused to the b subunit of vibrio cholera (ctb-ediii) was expressed in potatoes to develop a plant-based vaccine against dengue virus type 2. ctb-ediii fused to an endoplasmic reticulum (er) retention signal, sekdel, was introduced int ...201627335159
deploying aptameric sensing technology for rapid pandemic monitoring.the genome of virulent strains may possess the ability to mutate by means of antigenic shift and/or antigenic drift as well as being resistant to antibiotics with time. the outbreak and spread of these virulent diseases including avian influenza (h1n1), severe acute respiratory syndrome (sars-corona virus), cholera (vibrio cholera), tuberculosis (mycobacterium tuberculosis), ebola hemorrhagic fever (ebola virus) and aids (hiv-1) necessitate urgent attention to develop diagnostic protocols and as ...201626381238
an entamoeba histolytica adp-ribosyl transferase from the diphtheria toxin family modifies the bacterial elongation factor tu.adp-ribosyl transferases are enzymes involved in the post-translational modification of proteins; they participate in multiple physiological processes, pathogenesis and host-pathogen interactions. several reports have characterized the functions of these enzymes in viruses, prokaryotes and higher eukaryotes, but few studies have reported adp-ribosyl transferases in lower eukaryotes, such as parasites. the locus ehi_155600 from entamoeba histolytica encodes a hypothetical protein that possesses a ...201627234208
cationic antimicrobial peptides derived from crocodylus siamensis leukocyte extract, revealing anticancer activity and apoptotic induction on human cervical cancer cells.known antimicrobial peptides kt2 and rt2 as well as the novel rp9 derived from the leukocyte extract of the freshwater crocodile (crocodylus siamensis) were used to evaluate the ability in killing human cervical cancer cells. rp9 in the extract was purified by a combination of anion exchange column and reversed-phase hplc, and its sequence was analyzed by mass spectrometry. the novel peptide could inhibit gram-negative vibrio cholerae (clinical isolation) and gram-positive bacillus pumilus tistr ...201627129462
green synthesis and antimicrobial activity of silver nanoparticles using wild medicinal mushroom ganoderma applanatum (pers.) pat. from similipal biosphere reserve, odisha, india.in the present study, green synthesis and cost effective approach of silver nanoparticles using wild medicinal mushroom ganoderma applanatum (pers.) pat. from similipal biosphere reserve, odisha, india is reported. the biosynthesised agnps were characterised using uv-visible spectroscopy, particle analyser and scanning electron microscopy studies. it was found by dynamic light scattering analysis, that the average size and charges of the agnps were 133.0 ± 0.361 nm and -6.01 ± 5.30 mv, respectiv ...201627463787
hard-wired control of bacterial processes by chromosomal gene location.bacterial processes, such as stress responses and cell differentiation, are controlled at many different levels. while some factors, such as transcriptional regulation, are well appreciated, the importance of chromosomal gene location is often underestimated or even completely neglected. a combination of environmental parameters and the chromosomal location of a gene determine how many copies of its dna are present at a given time during the cell cycle. here, we review bacterial processes that r ...201627364121
use of a novel report protein to study the secretion signal of flagellin in bacillus subtilis.flagellin (also called hag) is the main component of bacterial flagellum and is transported across the cytoplasmic membrane by flagellar secretion apparatus. because flagella play an essential role in the pathogenesis of numerous pathogens, the flagellins of escherichia coli, salmonella typhimurium, pseudomonas aeruginosa, campylobacter jejuni, and vibrio cholerae have been intensively studied; however, very few studies have focused on the flagellin of bacillus subtilis, which is considered to b ...201627154466
antibacterial activity of an acidic phospholipase a2 (nn-xib-pla2) from the venom of naja naja (indian cobra).the resistance of bacteria against the use of conventional antibiotics has become a serious threat to public health and considering the associated side effect with antibiotics; new strategies to find and develop new molecules with novel modes of action has received grate attention in recent years. in this study, when the antibacterial potential of an acidic protein-nn-xib-pla2 (naja naja venom phospholipase a2 fraction-xib) of naja naja venom was evaluated, it showed significant bactericidal act ...201626885465
a specific antimicrobial protein cap-1 from pseudomonas sp. isolated from the jellyfish cyanea capillata.a bacterium strain, designated as cmf-2, was isolated from the jellyfish cyanea capillata and its culture supernatant exhibited a significant antimicrobial activity. the strain cmf-2 was identified as pseudomonas sp. based on the morphological, biochemical and physiological characteristics as well as 16s rrna sequence analysis. in this study, an antimicrobial protein, named as cap-1, was isolated from the culture of cmf-2 through ammonium sulfate precipitation and gel filtration chromatography. ...201626529191
bark extract mediated green synthesis of silver nanoparticles: evaluation of antimicrobial activity and antiproliferative response against osteosarcoma.in the current investigation we report the biosynthesis potentials of bark extracts of ficus benghalensis and azadirachta indica for production of silver nanoparticle without use of any external reducing or capping agent. the appearance of dark brown color indicated the complete nanoparticle synthesis which was further validated by absorbance peak by uv-vis spectroscopy. the morphology of the synthesized particles was characterized by field emission- scanning electron microscopy (fe-sem) and ato ...201626478285
the impact of oxygen on bacterial enteric pathogens.bacterial enteric pathogens are responsible for a tremendous amount of foodborne illnesses every year through the consumption of contaminated food products. during their transit from contaminated food sources to the host gastrointestinal tract, these pathogens are exposed and must adapt to fluctuating oxygen levels to successfully colonize the host and cause diseases. however, the majority of enteric infection research has been conducted under aerobic conditions. to raise awareness of the import ...201627261784
[monitoring and research on pathogen spectrum in patients with acute diarrhea from sentinel hospital of zhejiang province during 2009 to 2014].objective: to explore pathogen spectrum constitution of acute diarrhea in outpatient and emergency of zhejiang province, and provide basis for treatment, prevention and control of the disease. methods: during january 2009 to december 2014, we selected seven sentinel hospitals in different regions of zhejiang, monitored and researched on pathogen spectrum in patients with acute diarrhea from outpatient and emergency. we recorded patients' personal basic information, the main symptoms and signs, a ...201628057113
effects of ethyl acetate extracts from the polycephalomyces nipponicus isolate cod-mk1201 (ascomycetes) against human pathogenic bacteria and a breast cancer cell line.this study aimed to identify a suitable organic solvent for extracting bioactive compounds from polycephalomyces nipponicus and to evaluate the antibacterial and anticancer activities of the extracts obtained. only extracts obtained with ethyl acetate exhibited antibacterial activity, so ethyl acetate was chosen for large-scale extraction. the minimum inhibitory concentration (mic) and minimum bactericidal concentration (mbc) against gram-positive and gram-negative human pathogenic bacteria of t ...201627910791
case-control study of diarrheal disease etiology in individuals over 5 years in southwest china.acute diarrhea is one of the major public health problems worldwide. most of studies on acute diarrhea have been made on infants aged below 5 years and few efforts have been made to identify the etiological agents of acute diarrhea in people over five, especially in china.201627891182
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