Publications

TitleAbstractYear
Filter
PMID(sorted descending)
Filter
an optimized swine dysentery murine model to characterize shedding and clinical disease associated with "brachyspira hampsonii" infection.the development of a mouse model as an in vivo pathogenicity screening tool for brachyspira spp. has advanced the study of these economically important pathogens in recent years. however, none of the murine models published to date have been used to characterize the clinical signs of disease in mice, instead focusing on pathology following oral inoculation with various brachyspira spp. the experiments described herein explore modifications of published models to characterize faecal consistency, ...201728830508
a review of methods used for studying the molecular epidemiology of brachyspira hyodysenteriae.brachyspira (b.) spp. are intestinal spirochaetes isolated from pigs, other mammals, birds and humans. in pigs, seven brachyspira spp. have been described, i.e. b. hyodysenteriae, b. pilosicoli, b. intermedia, b. murdochii, b. innocens, b. suanatina and b. hampsonii. brachyspira hyodysenteriae is especially relevant in pigs as it causes swine dysentery and hence considerable economic losses to the pig industry. furthermore, reduced susceptibility of b. hyodysenteriae to antimicrobials is of incr ...201728757022
presence and mechanisms of acquired antimicrobial resistance in belgian brachyspira hyodysenteriae isolates belonging to different clonal complexes.swine dysentery (sd) is an economically important disease for which antimicrobial treatment still occupies an important place to control outbreaks. however, acquired antimicrobial resistance is increasingly observed in brachyspira hyodysenteriae. in this study, the minimal inhibitory concentrations (mic) of six antimicrobial compounds for 30 recent belgian b. hyodysenteriae isolates were determined using a broth microdilution method. in addition, relevant regions of the 16s rrna, 23s rrna and th ...201728757011
metabolic response of porcine colon explants to in vitro infection by brachyspira hyodysenteriae: a leap into disease pathophysiology.swine dysentery caused by brachyspira hyodysenteriae is a production limiting disease in pig farming. currently antimicrobial therapy is the only treatment and control method available.201728706469
emergence of brachyspira species and strains: reinforcing the need for surveillance.this short review discusses the increasing complexity that has developed around the understanding of brachyspira species that infect pigs, and their ability to cause disease. it describes the recognition of new weakly haemolytic brachyspira species, and the growing appreciation that brachyspira pilosicoli and some other weakly haemolytic species may be pathogenic in pigs. it discusses swine dysentery (sd) caused by the strongly haemolytic brachyspira hyodysenteriae, particularly the cyclical nat ...201528694985
in vitro susceptibility of brachyspira hyodysenteriae to a commercial citrus fruit extract.brachyspira hyodysenteriae is the main etiological agent of swine dysentery (sd). nowadays, treatment and control of sd is increasingly difficult due to the emergence of antimicrobial resistance together with the restrictions on the use of antibiotics in veterinary practice. the aim of this study was to evaluate, as an alternative in the control of this disease, the antimicrobial activity and the main mechanism of action of biocitro, a citrus extract commercialized as raw material and used as fe ...201728651094
neutrophil elastase and il17 expressed in the pig colon during brachyspira hyodysenteriae infection synergistically with the pathogen induce increased mucus transport speed and production via mapk3.brachyspira hyodysenteriae colonizes the pig colon resulting in mucoid hemorragic diarrhea and mucus layer changes. these changes are characterized by a disorganized mucus structure and a massive mucus induction with de novo expression of muc5ac and increased production of muc2. to investigate the mechanisms behind this altered mucin environment, we quantified the mrna levels of mucin pathway genes and factors from the immune system in the colon of infected and control pigs, revealing upregulati ...201728559407
brachyspira hyodysenteriae and b. pilosicoli proteins recognized by sera of challenged pigs.the spirochetes brachyspira hyodysenteriae and b. pilosicoli are pig intestinal pathogens that are the causative agents of swine dysentery (sd) and porcine intestinal spirochaetosis (pis), respectively. although some inactivated bacterin and recombinant vaccines have been explored as prophylactic treatments against these species, no effective vaccine is yet available. immunoproteomics approaches hold the potential for the identification of new, suitable candidates for subunit vaccines against sd ...201728522991
characterization of brachyspira hyodysenteriae isolates from italy by multilocus sequence typing and multiple locus variable number tandem repeat analysis.to evaluate and compare the capabilities of multilocus sequence typing (mlst) and multiple locus variable number tandem repeat analysis (mlva) techniques to characterize brachyspira hyodysenteriae isolates and to investigate the relationship between pleuromutilin resistance and genetic variability.201728510989
brachyspira hyodysenteriae infection regulates mucin glycosylation synthesis inducing an increased expression of core-2 o-glycans in porcine colon.brachyspira hyodysenteriae causes swine dysentery (sd), leading to global financial losses to the pig industry. infection with this pathogen results in an increase in b. hyodysenteriae binding sites on mucins, along with increased colonic mucin secretion. we predict that b. hyodysenteriae modifies the glycosylation pattern of the porcine intestinal mucus layer to optimize its host niche. we characterized the swine colonic mucin o-glycome and identified the differences in glycosylation between b. ...201728301166
an update of brachyspira hyodysenteriae serotyping.brachyspira (b.) hyodysenteriae the causative agent of swine dysentery (sd) has been divided into 9 serotypes on basis of its lipooligosaccharide (los). knowledge on circulating serotypes in europe, however, is rare. regarding that immunity to sd is serotype specific an update of b. hyodysenteriae serotyping was undertaken. a los band of 10 to 25kda was identified being appropriate for this purpose. isolates from germany, spain, denmark, usa and japan were characterized in the immunoblot by sera ...201728249175
artificially designed recombinant protein composed of multiple epitopes of foot-and-mouth disease virus as a vaccine candidate.concerns regarding the safety of inactivated foot-and-mouth disease (fmd) vaccine have been raised since it is produced from cultured live fmd virus (fmdv). to overcome this issue, recombinant protein has been studied as an alternative vaccine.201728228147
a novel multiplex qpcr targeting 23s rdna for diagnosis of swine dysentery and porcine intestinal spirochaetosis.a multiplex qpcr targeting a 128 bp region on the 23s rdna gene was developed for detection of brachyspira (b.) hyodysenteriae and b. pilosicoli, the agents of swine dysentery (sd) and porcine intestinal spirochaetosis (pis), together with a triplet of apathogenic brachyspira spp. (b. innocens, b. intermedia, b. murdochii) in porcine feces. the multiplex qpcr was evaluated against a duplex pcr (la et al., j clin microbiol 41:3372-5, 2003).201728173799
correction for mirajkar et al., complete genome sequence of brachyspira hyodysenteriae type strain b78 (atcc 27164). 201728104652
mechanisms of antibacterial action of quinoxaline 1,4-di-n-oxides against clostridium perfringens and brachyspira hyodysenteriae.quinoxaline 1,4-di-n-oxides (qdnos) are a class of bioreductive compounds, however, their antibacterial mechanisms are still unclarified. the aim of this study was to assess the ability of two representative qdno drugs, cyadox (cya) and olaquindox (ola), to produce reactive oxide species (ros) in gram-positive anaerobe clostridium perfringens cvcc1125 and gram-negative anaerobe brachyspira hyodysenteriae b204. in addition, the effects of qdnos on the integrity of bacterial cell walls and membran ...201628018297
the in vitro effect of garden thyme (thymus vulgaris l.) extract on brachyspira hyodysenteriae.filtrates of a water extract of commercially available garden thyme (thymus vulgaris l.) were used for studying its possible bactericidal effect on brachyspira hyodysenteriae, the causative agent of swine dysentery, by agar-diffusion technique. five of the six studied brachyspira strains have proven to be sensitive and one moderately sensitive in the in vitro tests. it was concluded that water extract of garden thyme possesses inhibitory effects against b. hyodysenteriae. in vivo experiments are ...201627929352
an investigation into the etiological agents of swine dysentery in australian pig herds.swine dysentery (sd) is a mucohemorrhagic colitis, classically seen in grower/finisher pigs and caused by infection with the anaerobic intestinal spirochete brachyspira hyodysenteriae. more recently, however, the newly described species brachyspira hampsonii and brachyspira suanatina have been identified as causing sd in north america and/or europe. furthermore, there have been occasions where strains of b. hyodysenteriae have been recovered from healthy pigs, including in multiplier herds with ...201627907102
toxigenicity of treponema hyodysenteriae, another emerging enteropathogen. 198227709524
the novel aminomethylcycline omadacycline has high specificity for the primary tetracycline-binding site on the bacterial ribosome.omadacycline is an aminomethylcycline antibiotic with potent activity against many gram-positive and gram-negative pathogens, including strains carrying the major efflux and ribosome protection resistance determinants. this makes it a promising candidate for therapy of severe infectious diseases. omadacycline inhibits bacterial protein biosynthesis and competes with tetracycline for binding to the ribosome. its interactions with the 70s ribosome were, therefore, analyzed in great detail and comp ...201627669321
characterization and recognition of brachyspira hampsonii sp. nov., a novel intestinal spirochete that is pathogenic to pigs.swine dysentery (sd) is a mucohemorrhagic colitis of swine classically caused by infection with the intestinal spirochete brachyspira hyodysenteriae since around 2007, cases of sd have occurred in north america associated with a different strongly beta-hemolytic spirochete that has been molecularly and phenotypically characterized and provisionally named "brachyspira hampsonii." despite increasing international interest, b. hampsonii is currently not recognized as a valid species. to support its ...201627629903
in vitro antimicrobial activities of animal-used quinoxaline 1,4-di-n-oxides against mycobacteria, mycoplasma and fungi.the quinoxaline 1,4-di-n-oxides (qdnos) were known as potent antibacterial agents. for the purpose of evaluating the bioactivity of existing animal-used qdnos drugs against representative pathogenic microorganism, the representative drugs of quinoxalines including cyadox, mequindox, quinocetone and their metabolites were submitted to the in vitro evaluation for antituberculosis, antimycoplasma, antifungal and antiviral activities.201627600955
complete genome sequence of brachyspira hyodysenteriae type strain b-78 (atcc 27164).reported herein is the complete genome sequence of the type strain b-78 (atcc 27164) of brachyspira hyodysenteriae, the etiological agent of swine dysentery. the 3.1-mb genome consists of a 3.056-mb chromosome and a 45-kb plasmid, with 2,617 protein-coding genes, 39 rna genes, and 40 pseudogenes.201627540064
understanding the culture of antimicrobial prescribing in agriculture: a qualitative study of uk pig veterinary surgeons.the use of antimicrobials in food-producing animals has been linked with the emergence of antimicrobial resistance in bacterial populations, with consequences for animal and public health. this study explored the underpinning drivers, motivators and reasoning behind prescribing decisions made by veterinary surgeons working in the uk pig industry.201627516473
antimicrobial susceptibility of brachyspira hyodysenteriae in switzerland.brachyspira (b.) hyodysenteriae is the causative agent of swine dysentery (sd), a severe mucohaemorrhagic diarrheal disease in pigs worldwide. so far, the antimicrobial susceptibility patterns of b. hyodysenteriae in switzerland have not been investigated. therefore, a panel of 30 porcine b. hyodysenteriae isolates were tested against 6 antimicrobial agents by using the vetmic brachy panel, a broth microdilution test. tiamulin and valnemulin showed high antimicrobial activity inhibiting all isol ...201627504836
the exposed proteomes of brachyspira hyodysenteriae and b. pilosicoli.brachyspira hyodysenteriae and brachyspira pilosicoli are well-known intestinal pathogens in pigs. b. hyodysenteriae is the causative agent of swine dysentery, a disease with an important impact on pig production while b. pilosicoli is responsible of a milder diarrheal disease in these animals, porcine intestinal spirochetosis. recent sequencing projects have provided information for the genome of these species facilitating the search of vaccine candidates using reverse vaccinology approaches. h ...201627493641
comparison of brachyspira hyodysenteriae isolates recovered from pigs in apparently healthy multiplier herds with isolates from herds with swine dysentery.swine dysentery (sd) is a mucohaemorrhagic colitis of grower/finisher pigs classically resulting from infection by the anaerobic intestinal spirochaete brachyspira hyodysenteriae. this study aimed to determine whether b. hyodysenteriae isolates from pigs in three healthy german multiplier herds supplying gilts to other farms differed from isolates from nine german production herds with sd. isolates were subjected to whole genomic sequencing, and in silico multilocus sequence typing showed that t ...201627489956
pipeline for amplifying and analyzing amplicons of the v1-v3 region of the 16s rrna gene.profiling of 16s rrna gene sequences is an important tool for testing hypotheses in complex microbial communities, and analysis methods must be updated and validated as sequencing technologies advance. in host-associated bacterial communities, the v1-v3 region of the 16s rrna gene is a valuable region to profile because it provides a useful level of taxonomic resolution; however, use of illumina miseq data for experiments targeting this region needs validation.201627485508
brachyspira hyodysenteriae isolated from apparently healthy pig herds following an evaluation of a prototype commercial serological elisa.swine dysentery (sd) is a disease mainly of grower/finisher pigs characterised by severe mucohaemorrhagic colitis. the classical aetiological agent is the anaerobic intestinal spirochaete brachyspira hyodysenteriae, although "brachyspira hampsonii" and brachyspira suanatina also cause sd. this study reports on the unexpected isolation of b. hyodysenteriae from pigs in apparently healthy herds that gave positive reactions when tested with a prototype commercial serological elisa for detecting her ...201627374902
functional and evolutionary characterization of a gene transfer agent's multilocus "genome".gene transfer agents (gtas) are phage-like particles that can package and transfer a random piece of the producing cell's genome, but are unable to transfer all the genes required for their own production. as such, gtas represent an evolutionary conundrum: are they selfish genetic elements propagating through an unknown mechanism, defective viruses, or viral structures "repurposed" by cells for gene exchange, as their name implies? in rhodobacter capsulatus, production of the r. capsulatus gta ( ...201627343288
variation in hemolytic activity of brachyspira hyodysenteriae strains from pigs.brachyspira hyodysenteriae is the primary cause of swine dysentery, which is responsible for major economic losses to the pig industry worldwide. the hemolytic activity of 10 b. hyodysenteriae strains isolated from stools of pigs with mild to mucohemorrhagic diarrhea was compared and seven hemolysis associated genes were sequenced. hemolysis induced by these strains varied from strong to near absent. one weakly hemolytic b. hyodysenteriae strain showed sequence changes in five hemolysis associat ...201627338265
swine dysentery.swine dysentery is a severe enteric disease in pigs, which is characterized by bloody to mucoid diarrhea and associated with reduced growth performance and variable mortality. this disease is most often observed in grower-finisher pigs, wherein susceptible pigs develop a significant mucohemorrhagic typhlocolitis following infection with strongly hemolytic spirochetes of the genus brachyspira. while swine dysentery is endemic in many parts of the world, the disease had essentially disappeared in ...201727288432
antimicrobial susceptibility patterns of brachyspira species isolated from swine herds in the united states.outbreaks of swine dysentery, caused by brachyspira hyodysenteriae and the recently discovered "brachyspira hampsonii," have reoccurred in north american swine herds since the late 2000s. additionally, multiple brachyspira species have been increasingly isolated by north american diagnostic laboratories. in europe, the reliance on antimicrobial therapy for control of swine dysentery has been followed by reports of antimicrobial resistance over time. the objectives of our study were to determine ...201627252458
effect of soy on faecal dry matter content and excretion of brachyspira hyodysenteriae in pigs.the aim of this study was to investigate the effect of a soy diet on the excretion of brachyspira hyodysenteriae in five farms with subclinically infected pigs. the effects on general health, faecal consistency and dry matter were analysed. in total, 200 pigs of different ages (group 1 <100 days of age (n=120) and group 2 ≥100 days (n=80)) were randomly assigned to the control (c) and the treatment (t) groups. group c received the farm's standard diet. in group t half of the daily feed ration wa ...201627239320
quinoxaline 1,4-di-n-oxides: biological activities and mechanisms of actions.quinoxaline 1,4-di-n-oxides (qdnos) have manifold biological properties, including antimicrobial, antitumoral, antitrypanosomal and antiinflammatory/antioxidant activities. these diverse activities endow them broad applications and prospects in human and veterinary medicines. as qdnos arouse widespread interest, the evaluation of their medicinal chemistry is still in progress. in the meantime, adverse effects have been reported in some of the qdno derivatives. for example, genotoxicity and bacte ...201627047380
dissemination of antimicrobial resistance in microbial ecosystems through horizontal gene transfer.the emergence and spread of antibiotic resistance among pathogenic bacteria has been a rising problem for public health in recent decades. it is becoming increasingly recognized that not only antibiotic resistance genes (args) encountered in clinical pathogens are of relevance, but rather, all pathogenic, commensal as well as environmental bacteria-and also mobile genetic elements and bacteriophages-form a reservoir of args (the resistome) from which pathogenic bacteria can acquire resistance vi ...201626925045
prevalence and risk factors for lawsonia intracellularis, brachyspira hyodysenteriae and salmonella spp. in finishing pigs in polish farrow-to-finish swine herds.the aim of the study was to estimate the herd-level, within-herd prevalence, the frequency of mixed infections and risk factors for l. intracellularis, b. hyodysenteriae and salmonella spp. in selected farrow-to-finish polish pig herds. a total of 254 pooled fecal samples were collected from 9 to 24 week-old pigs in 70 herds. real time pcr for detection of l. intracellularis and b. hyodysenteriae was performed. for salmonella spp. bacteriological examination was performed. the herd-level prevale ...201526812826
occurrence of brachyspira hyodysenteriae in multiplier pig herds in switzerland.this research was aimed to determine the occurrence of brachyspira (b.) hyodysenteriae in swiss multiplier pig herds.201626757784
evaluating the risk of pathogen transmission from wild animals to domestic pigs in australia.wild animals contribute to endemic infection in livestock as well as the introduction, reintroduction and maintenance of pathogens. the source of introduction of endemic diseases to a piggery is often unknown and the extent of wildlife contribution to such local spread is largely unexplored. the aim of the current study was to quantitatively assess the probability of domestic pigs being exposed to different pathogens from wild animals commonly found around commercial piggeries in australia. spec ...201626711303
evaluation of the in vitro activity of flumequine against field isolates of brachyspira hyodysenteriae.flumequine is a quinolone derivative used in veterinary medicine to treat enteric infections, mainly those caused by gram negative bacteria and also some gram positive. some recent reports by field practitioners have suggested that its use in swine dysentery outbreaks can minimize the impact of this disease. this study aims to evaluate the in vitro anti-brachyspira hyodysenteriae activity of flumequine. forty eight field isolates of the bacterium were evaluated using a microdilution test. the la ...201526679795
brachyspira pilosicoli-induced avian intestinal spirochaetosis.avian intestinal spirochaetosis (ais) is a common disease occurring in poultry that can be caused by brachyspira pilosicoli, a gram-negative bacterium of the order spirochaetes. during ais, this opportunistic pathogen colonises the lower gastrointestinal (gi) tract of poultry (principally, the ileum, caeca, and colon), which can cause symptoms such as diarrhoea, reduced growth rate, and reduced egg production and quality. due to the large increase of bacterial resistance to antibiotic treatment, ...201526679774
prevalence of chlamydial infections in fattening pigs and their influencing factors.chlamydial infections in pigs are associated with respiratory disease, diarrhea, conjunctivitis and other pathologies. the aim of this study was to define the prevalence of chlamydiaceae in swiss fattening pigs by applying sensitive and specific detection methods and to correlate prior antibiotic treatment and farm related factors with differences in prevalence. conjunctival and fecal swabs were collected from 636 pigs in 29 swiss fattening pig farms with and without antibiotic treatment, at the ...201526619187
drug-susceptibility of isolates of brachyspira hyodysenteriae isolated from colonic mucosal specimens of pigs collected from slaughter houses in japan in 2009.twenty nine isolates identified as brachyspira hyodysenteriae were most susceptible to carbadox and metronidazole, whereas they were resistant to macrolides. the isolates showed intermediate susceptibility to tiamulin, lincomycin, penicillin g, ampicillin, chloramphenicol, tetracycline, enrofloxacin and valnemulin, with mic50 values ranging from 0.39 to 3.13.201526596637
drug-susceptibility of isolates of brachyspira hyodysenteriae isolated from colonic mucosal specimens of pigs collected from slaughter houses in japan in 2009.twenty nine isolates identified as brachyspira hyodysenteriae were most susceptible to carbadox and metronidazole, whereas they were resistant to macrolides. the isolates showed intermediate susceptibility to tiamulin, lincomycin, penicillin g, ampicillin, chloramphenicol, tetracycline, enrofloxacin and valnemulin, with mic50 values ranging from 0.39 to 3.13.201526596637
porcine epidemic diarrhea virus among farmed pigs, ukraine.an outbreak of porcine epidemic diarrhea occurred in the summer of 2014 in ukraine, severely affecting piglets <10 days of age; the mortality rate approached 100%. full genome sequencing showed the virus to be closely related to strains reported from north america, showing a sequence identity of up to 99.8%.201526584081
alterations in the colonic microbiota of pigs associated with feeding distillers dried grains with solubles.in an effort to reduce feed costs, many pork producers have increased their use of coproducts of biofuel production in commercial pig diets, including increased feeding of distiller's dried grains with solubles (ddgs). the inclusion of ddgs increases the insoluble fiber content in the ration, which has the potential to impact the colonic microbiota considerably as the large intestine contains a dynamic microenvironment with tremendous interplay between microorganisms. any alteration to the physi ...201526555787
development of a serological elisa using a recombinant protein to identify pig herds infected with brachyspira hyodysenteriae.brachyspira hyodysenteriae is an anaerobic spirochaete that can induce swine dysentery (sd), a severe mucohaemorrhagic colitis in grower and fattener pigs. the aim of this study was to develop a serological elisa for use as a screening method to detect evidence of herd infection. bioinformatic analysis of the complete genome sequence of strain wa1 was used to identify genes predicted to encode outer membrane proteins. twenty candidate genes were expressed in an escherichia coli mediated system, ...201526542367
spirochetal motility and chemotaxis in the natural enzootic cycle and development of lyme disease.two-thirds of all bacterial genomes sequenced to-date possess an organelle for locomotion, referred to as flagella, periplasmic flagella or type iv pili. these genomes may also contain a chemotaxis-signaling system which governs flagellar rotation, thus leading a coordinated function for motility. motility and chemotaxis are often crucial for infection or disease process caused by pathogenic bacteria. although motility-associated genes are well-characterized in some organisms, the highly orchest ...201526519910
peptide/protein vaccine delivery system based on plga particles.due to the excellent safety profile of poly (d,l-lactide-co-glycolide) (plga) particles in human, and their biodegradability, many studies have focused on the application of plga particles as a controlled-release vaccine delivery system. antigenic proteins/peptides can be encapsulated into or adsorbed to the surface of plga particles. the gradual release of loaded antigens from plga particles is necessary for the induction of efficient immunity. various factors can influence protein release rate ...201526513024
peptide/protein vaccine delivery system based on plga particles.due to the excellent safety profile of poly (d,l-lactide-co-glycolide) (plga) particles in human, and their biodegradability, many studies have focused on the application of plga particles as a controlled-release vaccine delivery system. antigenic proteins/peptides can be encapsulated into or adsorbed to the surface of plga particles. the gradual release of loaded antigens from plga particles is necessary for the induction of efficient immunity. various factors can influence protein release rate ...201526513024
crystallographic characterization of the ribosomal binding site and molecular mechanism of action of hygromycin a.hygromycin a (hyga) binds to the large ribosomal subunit and inhibits its peptidyl transferase (pt) activity. the presented structural and biochemical data indicate that hyga does not interfere with the initial binding of aminoacyl-trna to the a site, but prevents its subsequent adjustment such that it fails to act as a substrate in the pt reaction. structurally we demonstrate that hyga binds within the peptidyl transferase center (ptc) and induces a unique conformation. specifically in its ribo ...201526464437
brachyspira suanatina sp. nov., an enteropathogenic intestinal spirochaete isolated from pigs and mallards: genomic and phenotypic characteristics.the genus brachyspira currently encompasses seven valid species that colonize the intestines of mammals and birds. in a previous study a group of strongly haemolytic isolates from pigs and mallards was provisionally described as a new species within genus brachyspira, "b. suanatina", and enteropathogenic properties were demonstrated in a porcine challenge model.201526458507
draft genome sequence of 'treponema phagedenis' strain v1, isolated from bovine digital dermatitis.'treponema phagedenis' is considered to be a key agent in the pathogenesis of bovine digital dermatitis, an infectious foot condition of economic and animal welfare importance. we hereby report the draft sequence of 't. phagedenis' strain v1. the draft genome assembly consists of 51 scaffolds comprising 3,129,551 bp and a gc-content of 39.9 %. putative pathogenicity related factors have been identified in the genome that can be used in future studies to gain insight into the pathogenic mechanism ...201526392840
lawsonia intracellularis infection of intestinal crypt cells is associated with specific depletion of secreted muc2 in goblet cells.the expression patterns of secreted (muc2 and muc5ac) and membrane-tethered (muc1, muc4, muc12 and muc13) mucins were monitored in healthy pigs and pigs challenged orally with lawsonia intracellularis. these results showed that the regulation of mucin gene expression is distinctive along the gi tract of the healthy pig, and may reflect an association between the function of the mucin subtypes and different physiological demands at various sites. we identified a specific depletion of secreted muc ...201526377360
in vitro susceptibility of brachyspira hyodysenteriae to organic acids and essential oil components.the antibacterial potential of organic acids and essential oil components against brachyspira hyodysenteriae, the causative pathogen of swine dysentery, was evaluated. minimum inhibitory concentrations (mic) of 15 compounds were determined at ph 7.2 and ph 6.0, using a broth microdilution assay. in addition, possible synergism was determined. mic values for the three tested strains were similar. for organic acids, mic values at ph 6.0 were lower than at ph 7.2. b. hyodysenteriae was most sensiti ...201526369432
in vitro susceptibility of brachyspira hyodysenteriae to organic acids and essential oil components.the antibacterial potential of organic acids and essential oil components against brachyspira hyodysenteriae, the causative pathogen of swine dysentery, was evaluated. minimum inhibitory concentrations (mic) of 15 compounds were determined at ph 7.2 and ph 6.0, using a broth microdilution assay. in addition, possible synergism was determined. mic values for the three tested strains were similar. for organic acids, mic values at ph 6.0 were lower than at ph 7.2. b. hyodysenteriae was most sensiti ...201526369432
peptide vaccine: progress and challenges.conventional vaccine strategies have been highly efficacious for several decades in reducing mortality and morbidity due to infectious diseases. the bane of conventional vaccines, such as those that include whole organisms or large proteins, appear to be the inclusion of unnecessary antigenic load that, not only contributes little to the protective immune response, but complicates the situation by inducing allergenic and/or reactogenic responses. peptide vaccines are an attractive alternative st ...201426344743
release and cytokine production of bmpb from bmpb-loaded ph-sensitive and mucoadhesive thiolated eudragit microspheres.swine dysentery is a contagious mucohaemorrhagic colitis of pigs that is caused by anaerobic intestinal spirochaete brachyspira hyodysenteriae. recently, an outer membrane lipoprotein of b. hyodysenteriae (bmpb) has been identified, and the mice or pigs immunized with a recombinant bmpb generated antibodies recognizing the native bmpb of b. hyodysenteriae. in this study, we cloned, expressed and purified bmpb protein from e. coli and used it as a vaccine candidate for oral delivery. the bmpb was ...201526328412
the altered schaedler flora: continued applications of a defined murine microbial community.the gastrointestinal (gi) microbiota forms a mutualistic relationship with the host through complex and dynamic interactions. because of the complexity and interindividual variation of the gi microbiota, investigating how members of the microbiota interact with each other, as well as with the host, is daunting. the altered schaedler flora (asf) is a model community of eight microorganisms that was developed by r.p. orcutt and has been in use since the late 1970s. the eight microorganisms composi ...201526323627
the gastrointestinal tract as a potential infection reservoir of digital dermatitis-associated treponemes in beef cattle and sheep.digital dermatitis (dd) is an important cause of lameness in dairy cattle worldwide. it has now been reported in beef cattle and also sheep (contagious ovine digital dermatitis [codd]). three treponema phylogroups are consistently isolated from lesions, treponema medium-like, treponema phagedenis-like, and treponema pedis. the gastrointestinal (gi) tract and feces are suggested sites of treponemal infection in dairy cattle; however, isolation of dd-associated treponemes from these areas has prev ...201526276110
risk assessment as a tool for improving external biosecurity at farm level.biosecurity routines at herd level may reduce the probability of introduction of disease into the herd, but some measures may be regarded as expensive and cumbersome for the farmers. custom-made measures based on individual farm characteristics may aid in improving the actual application of on-farm biosecurity. the aim of the study was to provide a tool for calculating the effects of different biosecurity measures and strategies on the individual farm level. a simple model was developed to asses ...201526215281
complete genome sequence and description of salinispira pacifica gen. nov., sp. nov., a novel spirochaete isolated form a hypersaline microbial mat.during a study of the anaerobic microbial community of a lithifying hypersaline microbial mat of lake 21 on the kiritimati atoll (kiribati republic, central pacific) strain l21-rpul-d2(t) was isolated. the closest phylogenetic neighbor was spirochaeta africana z-7692(t) that shared a 16s rrna gene sequence identity value of 90% with the novel strain and thus was only distantly related. a comprehensive polyphasic study including determination of the complete genome sequence was initiated to chara ...201526203324
weaner production with low antimicrobial usage: a descriptive study.health, productivity and antimicrobial use in the production of pigs are expected to be interrelated to some extent. previous studies on register-based data have investigated these correlations with a subsequent large variation residing at the farm level. in order to study such farm factors in more detail we designed an elaborate interview-guide. by in-depth interviews of farmers with well-managed 7-30 kg (weaner) productions we sought to describe a set of common key-factors characterizing their ...201526183944
molecular epidemiology of novel pathogen "brachyspira hampsonii" reveals relationships between diverse genetic groups, regions, host species, and other pathogenic and commensal brachyspira species.outbreaks of bloody diarrhea in swine herds in the late 2000s signaled the reemergence of an economically significant disease, swine dysentery, in the united states. investigations confirmed the emergence of a novel spirochete in swine, provisionally designated "brachyspira hampsonii," with two genetically distinct clades. although it has since been detected in swine and migratory birds in europe and north america, little is known about its genetic diversity or its relationships with other brach ...201526135863
analysis of multiple brachyspira hyodysenteriae genomes confirms that the species is relatively conserved but has potentially important strain variation.the intestinal spirochete brachyspira hyodysenteriae is an important pathogen in swine, causing mucohemorrhagic colitis in a disease known as swine dysentery. based on the detection of significant linkage disequilibrium in multilocus sequence data, the species is considered to be clonal. an analysis of the genome sequence of western australian b. hyodysenteriae strain wa1 has been published, and in the current study 19 further strains from countries around the world were sequenced with illumina ...201526098837
heterocyclic n-oxides - an emerging class of therapeutic agents.heterocyclic n-oxides have emerged as potent compounds with anticancer, antibacterial, antihypertensive, antiparasitic, anti-hiv, anti-inflammatory, herbicidal, neuroprotective, and procognitive activities. the n-oxide motif has been successfully employed in a number of recent drug development projects. this review surveys the emergence of this scaffold in the mainstream medicinal chemistry with a focus on the discovery of the heterocyclic n-oxide drugs, n-oxide-specific mechanisms of action, dr ...201526087764
clostridium butyricum strains and dysbiosis linked to necrotizing enterocolitis in preterm neonates.necrotizing enterocolitis (nec) is the most common and serious gastrointestinal disorder among preterm neonates. we aimed to assess a specific gut microbiota profile associated with nec.201526084844
distinct trna accommodation intermediates observed on the ribosome with the antibiotics hygromycin a and a201a.the increase in multi-drug-resistant bacteria is limiting the effectiveness of currently approved antibiotics, leading to a renewed interest in antibiotics with distinct chemical scaffolds. we have solved the structures of the thermus thermophilus 70s ribosome with a-, p-, and e-site trnas bound and in complex with either the aminocyclitol-containing antibiotic hygromycin a (hyga) or the nucleoside antibiotic a201a. both antibiotics bind at the peptidyl transferase center and sterically occlude ...201526028538
investigation of biosecurity risks associated with the feed delivery: a pilot study.this study explored potential biosecurity issues related to the delivery of feed to commercial farms. a pilot study was conducted to collect information about the day-to-day feed delivery, including biosecurity concerns at the level of the feed truck, the driver, and the farm. in addition, a reusable rubber boot was tested in an effort to increase the proportion of farms at which truck drivers wore clean footwear, and to explore an alternative to the standard plastic disposable boots that may be ...201525969585
prevalence and genetic diversity of clinical vibrio parahaemolyticus isolates from china, revealed by multilocus sequence typing scheme.the population structure of clinical vibrio parahaemolyticus isolates spreading in china remains undefined. we brought 218 clinical isolates from the pubmlst database originating from different regions of china collected since the year of 1990, analyzed by multilocus sequence typing (mlst), to elucidate the prevalence and genetic diversity of v. parahaemolyticus circulating in chinese population. the mlst scheme produced 137 sequence types (sts). these sts were clustered into six clonal complexe ...201525914691
characterization of a caprine model for the subclinical initial phase of mycobacterium avium subsp. paratuberculosis infection.paratuberculosis caused by mycobacterium avium subsp. paratuberculosis (map) is difficult to control due to a long phase of clinically non-apparent (latent) infection for which sensitive diagnostics are lacking. a defined animal model for this phase of the infection can help to investigate host-map interactions in apparently healthy animals and identify surrogate markers for disease progress and might also serve as challenge model for vaccines. to establish such a model in goats, different age a ...201525889716
biosecurity level and health management practices in 60 swedish farrow-to-finish herds.biosecurity measures are important tools to maintain animal health in pig herds. within the minapig project, whose overall aim is to evaluate strategies to raise pigs with less antimicrobial use, biosecurity was evaluated in medium to large farrow-to-finish pig herds in sweden. in 60 farrow-to-finish herds with more than 100 sows, the biosecurity level was evaluated using a previously developed protocol (biocheck). in a detailed questionnaire, internal and external biosecurity was scored in six ...201525887040
mycobacterium tuberculosis tlya protein negatively regulates t helper (th) 1 and th17 differentiation and promotes tuberculosis pathogenesis.mycobacterium tuberculosis, the causative agent of tuberculosis, is an ancient pathogen and a major cause of death worldwide. although various virulence factors of m. tuberculosis have been identified, its pathogenesis remains incompletely understood. tlya is a virulence factor in several bacterial infections and is evolutionarily conserved in many gram-positive bacteria, but its function in m. tuberculosis pathogenesis has not been elucidated. here, we report that tlya significantly contributes ...201525847237
mutations in the bacterial ribosomal protein l3 and their association with antibiotic resistance.different groups of antibiotics bind to the peptidyl transferase center (ptc) in the large subunit of the bacterial ribosome. resistance to these groups of antibiotics has often been linked with mutations or methylations of the 23s rrna. in recent years, there has been a rise in the number of studies where mutations have been found in the ribosomal protein l3 in bacterial strains resistant to ptc-targeting antibiotics but there is often no evidence that these mutations actually confer antibiotic ...201525845869
the levels of brachyspira hyodysenteriae binding to porcine colonic mucins differ between individuals, and binding is increased to mucins from infected pigs with de novo muc5ac synthesis.brachyspira hyodysenteriae colonizes the pig colon, resulting in mucohemorrhagic diarrhea and growth retardation. fecal mucus is a characteristic feature of swine dysentery; therefore, we investigated how the mucin environment changes in the colon during infection with b. hyodysenteriae and how these changes affect this bacterium's interaction with mucins. we isolated and characterized mucins, the main component of mucus, from the colon of experimentally inoculated and control pigs and investiga ...201525644008
the rebirth of culture in microbiology through the example of culturomics to study human gut microbiota.bacterial culture was the first method used to describe the human microbiota, but this method is considered outdated by many researchers. metagenomics studies have since been applied to clinical microbiology; however, a "dark matter" of prokaryotes, which corresponds to a hole in our knowledge and includes minority bacterial populations, is not elucidated by these studies. by replicating the natural environment, environmental microbiologists were the first to reduce the "great plate count anomal ...201525567229
in situ prebiotics for weaning piglets: in vitro production and fermentation of potato galacto-rhamnogalacturonan.postweaning diarrhea (pwd) in pigs is a leading cause of economic loss in pork production worldwide. the current practice of using antibiotics and zinc to treat pwd is unsustainable due to the potential of antibiotic resistance and ecological disturbance, and novel methods are required. in this study, an in vitro model was used to test the possibility of producing prebiotic fiber in situ in the gastrointestinal (gi) tract of the piglet and the prebiotic activity of the resulting fiber in the ter ...201525527557
comparison of culture, polymerase chain reaction, and fluorescent in situ hybridization for detection of brachyspira hyodysenteriae and "brachyspira hampsonii" in pig feces.swine dysentery is characterized by mucohemorrhagic diarrhea and can occur following infection by brachyspira hyodysenteriae or "brachyspira hampsonii ". a definitive diagnosis is often based on the isolation of strongly beta-hemolytic spirochetes from selective culture or by the application of species-specific polymerase chain reaction (pcr) assays directly to feces. while culture is highly sensitive, it typically requires 6 or more days to complete, and pcr, although rapid, can be limited by f ...201525525136
absence of a set of plasmid-encoded genes is predictive of reduced pathogenic potential in brachyspira hyodysenteriae.the gene content of 14 strains of the intestinal spirochaete brachyspira hyodysenteriae was compared using a dna microarray. a consistent difference occurred in a block of four genes on the ~36 kb plasmid, with these being present in six virulent strains and absent in eight strains with reduced pathogenic potential. these genes encoded a predicted radical s-adenosylmethionine domain protein, a glycosyl transferase group 1-like protein, an nad dependent epimerase and a dtdp-4-dehydrorhamnose 2-5 ...201425512147
sequence types and pleuromutilin susceptibility of brachyspira hyodysenteriae isolates from italian pigs with swine dysentery: 2003-2012.swine dysentery is a mucohaemorrhagic colitis of pigs caused by infection with brachyspira hyodysenteriae. the disease can be controlled by treatment with antimicrobial agents, with the pleuromutilins tiamulin and valnemulin being widely used. in recent years, the occurrence of b. hyodysenteriae with reduced susceptibility to these drugs has been increasing. the aim of this study was to determine temporal changes in genetic groups and pleuromutilin susceptibility amongst b. hyodysenteriae isolat ...201525486860
investigation of the impact of increased dietary insoluble fiber through the feeding of distillers dried grains with solubles (ddgs) on the incidence and severity of brachyspira-associated colitis in pigs.diet has been implicated as a major factor impacting clinical disease expression of swine dysentery and brachyspira hyodysenteriae colonization. however, the impact of diet on novel pathogenic strongly beta-hemolytic brachyspira spp. including "b. hampsonii" has yet to be investigated. in recent years, distillers dried grains with solubles (ddgs), a source of insoluble dietary fiber, has been increasingly included in diets of swine. a randomized complete block experiment was used to examine the ...201425485776
development of aminoglycoside and β-lactamase resistance among intestinal microbiota of swine treated with lincomycin, chlortetracycline, and amoxicillin.lincomycin, chlortetracycline, and amoxicillin are commonly used antimicrobials for growth promotion and infectious disease prophylaxis in swine production. in this study, we investigated the shifts and resistance development among intestinal microbiota in pregnant sows before and after lincomycin, chlortetracycline, and amoxicillin treatment by using phylogenetic analysis, bacterial enumeration, and pcr. after the antimicrobial treatment, shifts in microbial community, an increased proportion o ...201425408688
genome mining for ribosomally synthesized and post-translationally modified peptides (ripps) in anaerobic bacteria.ribosomally synthesized and post-translationally modified peptides (ripps) are a diverse group of biologically active bacterial molecules. due to the conserved genomic arrangement of many of the genes involved in their synthesis, these secondary metabolite biosynthetic pathways can be predicted from genome sequence data. to date, however, despite the myriad of sequenced genomes covering many branches of the bacterial phylogenetic tree, such an analysis for a broader group of bacteria like anaero ...201425407095
16s rrna gene mutations associated with decreased susceptibility to tetracycline in mycoplasma bovis.mycoplasma bovis isolates with decreased susceptibilities to tetracyclines are increasingly reported worldwide. the acquired molecular mechanisms associated with this phenomenon were investigated in 70 clinical isolates of m. bovis. sequence analysis of the two 16s rrna-encoding genes (rrs3 and rrs4 alleles) containing the primary binding pocket for tetracycline (tet-1 site) was performed on isolates with tetracycline hydrochloride mics of 0.125 to 16 μg/ml. mutations at positions a965t, a967t/c ...201525403668
herd diagnosis of low pathogen diarrhoea in growing pigs - a pilot study.the major indication for antibiotic use in danish pigs is treatment of intestinal diseases post weaning. clinical decisions on antibiotic batch medication are often based on inspection of diarrhoeic pools on the pen floor. in some of these treated diarrhoea outbreaks, intestinal pathogens can only be demonstrated in a small number of pigs within the treated group (low pathogen diarrhoea). termination of antibiotic batch medication in herds suffering from such diarrhoea could potentially reduce t ...201425392732
cessation of clinical disease and spirochete shedding after tiamulin treatment in pigs experimentally infected with "brachyspira hampsonii".with the emergence of "brachyspira hampsonii" associated with swine dysentery in north america, identification of effective treatments and interventions is a pressing need. denagard® (tiamulin hydrogen fumarate) liquid concentrate 12.5% is approved in the united states for treatment of dysentery caused by brachyspira hyodysenteriae at 0.006% in the water. in this study, the effectiveness of tiamulin in resolving clinical disease, eliminating viable spirochete shedding, and reducing neutrophilic ...201425218812
understanding the molecular epidemiology and global relationships of brachyspira hyodysenteriae from swine herds in the united states: a multi-locus sequence typing approach.outbreaks of mucohemorrhagic diarrhea in pigs caused by brachyspira hyodysenteriae in the late 2000s indicated the re-emergence of swine dysentery (sd) in the u.s. although the clinical disease was absent in the u.s. since the early 1990s, it continued to cause significant economic losses to other swine rearing countries worldwide. this study aims to fill the gap in knowledge pertaining to the re-emergence and epidemiology of b. hyodysenteriae in the u.s. and its global relationships using a mul ...201425192199
holins in bacteria, eukaryotes, and archaea: multifunctional xenologues with potential biotechnological and biomedical applications.holins form pores in the cytoplasmic membranes of bacteria for the primary purpose of releasing endolysins that hydrolyze the cell wall and induce cell death. holins are encoded within bacteriophage genomes, where they promote cell lysis for virion release, and within bacterial genomes, where they serve a diversity of potential or established functions. these include (i) release of gene transfer agents, (ii) facilitation of programs of differentiation such as those that allow sporulation and spo ...201425157079
holins in bacteria, eukaryotes, and archaea: multifunctional xenologues with potential biotechnological and biomedical applications.holins form pores in the cytoplasmic membranes of bacteria for the primary purpose of releasing endolysins that hydrolyze the cell wall and induce cell death. holins are encoded within bacteriophage genomes, where they promote cell lysis for virion release, and within bacterial genomes, where they serve a diversity of potential or established functions. these include (i) release of gene transfer agents, (ii) facilitation of programs of differentiation such as those that allow sporulation and spo ...201425157079
[eradication of swine dysentery as modified partial depopulation in a nucleus sow breeding farm].on a swiss nucleus sow breeding farm with 170 sows and 600 gilts/fatteners, an eradication of swine dysentery as modified partial depopulation was conducted in stages over a period of 12 weeks in 2011 after brachyspira (b.) hyodysenteriae was detected in the herd. in addition to administering oral medication (8.1 mg tiamulin per kg body weight) for 4 weeks to the pigs remaining on the farm, all stables were cleaned thoroughly and the residual slurry was disinfected with alzogur(®) (3 l/m(3)) whi ...201425082634
matrix-assisted laser desorption ionization time-of-flight mass spectrometry for rapid identification of brachyspira species isolated from swine, including the newly described "brachyspira hampsonii".the brachyspira species traditionally associated with swine dysentery and other diarrheal diseases in pigs are brachyspira hyodysenteriae, brachyspira pilosicoli, and, to a lesser extent, brachyspira murdochii. "brachyspira hampsonii" is a recently proposed novel species that causes clinical disease similar to that caused by b. hyodysenteriae. matrix-assisted laser desorption ionization time-of-flight mass spectrometry (maldi-tof ms) systems are increasingly available in veterinary diagnostic la ...201425012082
evolutionary origin of insect-wolbachia nutritional mutualism.obligate insect-bacterium nutritional mutualism is among the most sophisticated forms of symbiosis, wherein the host and the symbiont are integrated into a coherent biological entity and unable to survive without the partnership. originally, however, such obligate symbiotic bacteria must have been derived from free-living bacteria. how highly specialized obligate mutualisms have arisen from less specialized associations is of interest. here we address this evolutionary issue by focusing on an ex ...201424982177
carbadox has both temporary and lasting effects on the swine gut microbiota.antibiotics are used in livestock and poultry production to treat and prevent disease as well as to promote animal growth. carbadox is an in-feed antibiotic that is widely used in swine production to prevent dysentery and to improve feed efficiency. the goal of this study was to characterize the effects of carbadox and its withdrawal on the swine gut microbiota. six pigs (initially 3-weeks old) received feed containing carbadox and six received unamended feed. after 3-weeks of continuous carbado ...201424959163
mutations in the 50s ribosomal subunit of brachyspira hyodysenteriae associated with altered minimum inhibitory concentrations of pleuromutilins.brachyspira hyodysenteriae, the causative agent of swine dysentery, is responsible for severe mucohaemorrhagic colitis with considerable financial loss to worldwide swine production. antimicrobial resistance against macrolides and lincosamides is widespread and the mechanisms are well known. currently, the most common treatment for swine dysentery is the use of pleuromutilins and resistance to these drugs also is increasingly being reported. although resistance mechanisms against pleuromutilins ...201424948419
confirmation that "brachyspira hampsonii" clade i (canadian strain 30599) causes mucohemorrhagic diarrhea and colitis in experimentally infected pigs."brachyspira hampsonii", discovered in north america in 2010 associated with dysentery-like illness, is an economically relevant swine pathogen resulting in decreased feed efficiency and increased morbidity, mortality and medication usage. "b. hampsonii" clade ii strain 30446 has been shown to be causally associated with mucohemorrhagic diarrhea and colitis. our objectives were to determine if "brachyspira hampsonii" clade i strain 30599 is pathogenic to pigs, and to evaluate the relative diagno ...201424917084
estimating diagnostic test accuracies for brachyspira hyodysenteriae accounting for the complexities of population structure in food animals.for swine dysentery, which is caused by brachyspira hyodysenteriae infection and is an economically important disease in intensive pig production systems worldwide, a perfect or error-free diagnostic test ("gold standard") is not available. in the absence of a gold standard, bayesian latent class modelling is a well-established methodology for robust diagnostic test evaluation. in contrast to risk factor studies in food animals, where adjustment for within group correlations is both usual and re ...201424906140
down-regulation of mechanisms involved in cell transport and maintenance of mucosal integrity in pigs infected with lawsonia intracellularis.lawsonia intracellularis is an obligate intracellular bacterium, responsible for the disease complex known as proliferative enteropathy (pe). l. intracellularis is associated with intestinal crypt epithelial cell proliferation but the mechanisms responsible are yet to be defined. microarray analysis was used to investigate the host-pathogen interaction in experimentally infected pigs to identify pathways that may be involved. ileal samples originating from twenty-eight weaner pigs experimentally ...201424885874
antibiotic alternatives: the substitution of antibiotics in animal husbandry?it is a common practice for decades to use of sub-therapeutic dose of antibiotics in food-animal feeds to prevent animals from diseases and to improve production performance in modern animal husbandry. in the meantime, concerns over the increasing emergence of antibiotic-resistant bacteria due to the unreasonable use of antibiotics and an appearance of less novelty antibiotics have prompted efforts to develop so-called alternatives to antibiotics. whether or not the alternatives could really rep ...201424860564
pleuromutilins: use in food-producing animals in the european union, development of resistance and impact on human and animal health.pleuromutilins (tiamulin and valnemulin) are antimicrobial agents that are used mainly in veterinary medicine, especially for swine and to a lesser extent for poultry and rabbits. in pigs, tiamulin and valnemulin are used to treat swine dysentery, spirochaete-associated diarrhoea, porcine proliferative enteropathy, enzootic pneumonia and other infections where mycoplasma is involved. there are concerns about the reported increases in the mics of tiamulin and valnemulin for porcine brachyspira hy ...201424793902
seroprevalence of lawsonia intracellularis antibodies in intensive pig farms in china.porcine proliferative enteropathy caused by lawsonia intracellularis (l. intracellularis) is a major concern to the pig industry worldwide. although 8.3 billion pigs are produced each year in china, few reports on the prevalence of l.intracellularis infection are available. the aim of the current study was to estimate the seroprevalence of l. intracellularis antibodies in intensive pig farms in china.201424774304
a field evaluation of two vaccines against mycoplasma hyopneumoniae infection in pigs.a field trial was carried out with two mycoplasma hyopneumoniae vaccines in order to investigate the benefit of vaccination under field conditions in modern danish pig production facilities with pigs being positive for m. hyopneumoniae. the m. hyopneumoniae infection of the herd was confirmed through blood samples that were positive for antibodies against m. hyopneumoniae combined with gross lesions of the lungs related to m. hyopneumoniae at slaughter and detection of m. hyopneumoniae by polyme ...201424739629
Displaying items 1 - 100 of 818