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phenotypic and genotypic antimicrobial resistance profiles of campylobacter jejuni isolated from cattle, sheep, and free-range poultry faeces.minimum inhibitory concentrations (mic) of 13 antimicrobial agents were determined by broth microdilution for 72 campylobacter jejuni strains from livestock. twenty-three (31.9%) isolates were fully susceptible; all isolates were susceptible to erythromycin, chloramphenicol, streptomycin, gentamicin, sulfamethoxazole, and meropenem, and all but one to kanamycin. resistance to quinolones was highest (52.8%), reaching similar values among poultry, dairy cattle, and sheep, but lower in beef cattle. ...201020224816
epidemiologic investigation of immune-mediated polyradiculoneuropathy among abattoir workers exposed to porcine brain.in october 2007, a cluster of patients experiencing a novel polyradiculoneuropathy was identified at a pork abattoir (plant a). patients worked in the primary carcass processing area (warm room); the majority processed severed heads (head-table). an investigation was initiated to determine risk factors for illness.201020333310
genotyping and pcr detection of potential virulence genes in campylobacter jejuni and campylobacter coli isolates from different sources in poland.the prevalence of potential virulence markers was determined among the population of polish campylobacter jejuni and campylobacter coli isolates from children, chickens, pigs and dogs. the presence of the flaa, flab, cdta, cdtb, cdtc, cdtabc, virb11, and cj0588 genes among 74 c. jejuni and 15 c. coli isolates was detected by pcr. high prevalence of five different putative virulence and toxin genes (flaa, cdta, cdtb, cdtc, and cj0588) was found among isolates obtained from children, chickens and ...201020490760
multi-locus sequence types of campylobacter carried by flies and slugs acquired from local ruminant faeces.to assess whether flies and slugs acquire strains of campylobacter jejuni and campylobacter coli present in local ruminant faeces.201020337762
fluorescence in situ hybridization method using a peptide nucleic acid probe for identification of salmonella spp. in a broad spectrum of samples.a fluorescence in situ hybridization (fish) method for the rapid detection of salmonella spp. using a novel peptide nucleic acid (pna) probe was developed. the probe theoretical specificity and sensitivity were both 100%. the pna-fish method was optimized, and laboratory testing on representative strains from the salmonella genus subspecies and several related bacterial species confirmed the predicted theoretical values of specificity and sensitivity. the pna-fish method has been successfully ad ...201020453122
evolutionary dynamics of complete campylobacter pan-genomes and the bacterial species concept.defining bacterial species and understanding the relative cohesiveness of different components of their genomes remains a fundamental problem in microbiology. bacterial species tend to be comprised of both a set of core and dispensable genes, with the sum of these two components forming the species pan-genome. the role of the core and dispensable genes in defining bacterial species and the question of whether pan-genomes are finite or infinite remain unclear. here we demonstrate, through the ana ...201020688752
repeated therapeutic dosing selects macrolide-resistant campylobacter spp. in a turkey facility.this study assessed the effects of the therapeutic use of tylan® in a large-scale turkey production facility on the selection of macrolide-resistant campylobacter.201020497488
fluoroquinolone resistance detection in campylobacter coli and campylobacter jejuni by luminex xmap technology.the proportion of campylobacter spp. isolates that are resistant to fluoroquinolones, the drugs of choice for campylobacteriosis, has been increasing worldwide. we developed an innovative method based on a luminex xmap dna suspension array that allows the identification of campylobacter species and, simultaneously, the detection of the most common point mutation in the gyra gene (substitution from threonine 86 to isoleucine 86) that is responsible for fluoroquinolone resistance. ninety-six campy ...201020500084
prevalence and antimicrobial resistance of campylobacter jejuni and campylobacter coli isolated from cattle between 2002 and 2006 in france.feces from 2,255 cattle (calves, young beef cattle, and culled cows) were collected at slaughter from nine departments across france. campylobacter was recovered from 16.5% of the 2,255 samples (c. jejuni from 12.8% and c. coli from 3.7%), predominantly from calves. antimicrobial resistance to six antibiotics of medical and/or veterinary interest was tested with the e-test. resistance to tetracycline was found in most isolates (52.8% of c. jejuni isolates and 88.1% of c. coli isolates) in contra ...201020501032
obstacles of multiplex real-time pcr for bacterial 16s rdna: primer specifity and dna decontamination of taq polymerase.background: the detection of a broad range of bacteria by pcr is applied for the screening of blood and blood products with special attention to platelet concentrates. for practical use it is desirable that detection systems include gram-positive, gram-negative and non-gram-stainable bacteria. it is quite challenging to achieve high sensitivity along with a clear negative control with pcr reagents, because especially taq polymerase is contaminated with traces of bacterial dna. methods: bacterial ...201020737013
investigation of water washes suitable for very small meat plants to reduce pathogens on beef surfaces.water washing with a handheld hose was performed on beef surfaces to ascertain the most effective combination of methods needed to remove potentially harmful microorganisms. for these experiments, beef brisket surfaces were experimentally inoculated with a fecal slurry containing escherichia coli o157:h7, salmonella typhimurium, campylobacter coli, and campylobacter jejuni. in a pilot study, surfaces were washed with cold water (15 degrees c) at various water pressures, spray distances, applicat ...201020501042
identification of campylobacter jejuni genes involved in its interaction with epithelial cells.campylobacter jejuni is the leading cause of infectious gastroenteritis in industrialized nations. its ability to enter and survive within nonphagocytic cells is thought to be very important for pathogenesis. however, little is known about the c. jejuni determinants that mediate these processes. through an extensive transposon mutagenesis screen, we have identified several loci that are required for c. jejuni efficient entry and survival within epithelial cells. among these loci, insertional mut ...201020515930
three-hour molecular detection of campylobacter, salmonella, yersinia, and shigella species in feces with accuracy as high as that of culture.campylobacter jejuni and salmonella, shigella, and yersinia species (along with shiga toxin-producing escherichia coli) are the most common causes of acute bacterial diarrheal disease in the united states. current detection techniques are time-consuming, limiting usefulness for patient care. we developed and validated a panel of rapid pcr assays for the detection and identification of c. jejuni, c. coli, salmonella, and yersinia species and shigella and enteroinvasive e. coli in stool samples. a ...201020519461
host association of campylobacter genotypes transcends geographic variation.genetic attribution of bacterial genotypes has become a major tool in the investigation of the epidemiology of campylobacteriosis and has implicated retail chicken meat as the major source of human infection in several countries. to investigate the robustness of this approach to the provenance of the reference data sets used, a collection of 742 campylobacter jejuni and 261 campylobacter coli isolates obtained from united kingdom-sourced chicken meat was established and typed by multilocus seque ...201020525862
molecular typing of campylobacter jejuni and c. coli from chickens and patients with gastritis or guillain-barré syndrome based on multilocus sequence types and pulsed-field gel electrophoresis patterns.campylobacter jejuni has recently been noted as the most common cause of bacterial foodborne diseases in japan. in the present study, we determined st types of c. jejuni and campylobacter coli isolated from chickens and patients with enteritis or gbs in japan and thailand. c. jejuni from chickens, enteritis, and gbs exhibited divergent st types and included several novel types in addition to worldwide common types. c. coli from enteritis was also divergent. novel st types may represent unidentif ...201020536735
comparison of real-time pcr assays for detection, quantification, and differentiation of campylobacter jejuni and campylobacter coli in broiler neck skin samples.we tested the use of multiplex real-time pcr for detection and quantification of campylobacter jejuni and campylobacter coli on broiler carcass neck skin samples collected during 2008 from slaughterhouses in switzerland. results from an established taqman assay based on two different targets (hipo and ceue for c. jejuni and c. coli, respectively) were corroborated with data from a newly developed assay based on a single-nucleotide polymorphism in the fusa gene, which allows differentiation betwe ...201020537260
screening for several potential pathogens in feral pigeons (columba livia) in madrid.pathogens with the zoonotic potential to infect humans, such as campylobacter jejuni, campylobacter coli and chlamydophila psittaci, can be found in feral pigeons (columba livia). given the high density of these birds in the public parks and gardens of most cities, they may pose a direct threat to public health.201020569487
differences in methylation at gatc sites in genomic dna of campylobacter coli from turkeys and swine.a significant fraction (46/108, 43%) of swine isolates of campylobacter coli but none of 81 isolates of c. coli from turkeys had genomic dna that was resistant to digestion by mboi, suggesting methylation of adenines at gatc sites. no consistent association was noted between antimicrobial resistance and mboi resistance. seven swine-associated multilocus sequence typing-based sequence types (sts) were detected among multiple isolates with mboi-resistant dna. the data suggest host-associated dna m ...201020833787
prevalence of virulence-associated genes and cytolethal distending toxin production in campylobacter spp. isolated in italy.the prevalence of virulence and cytolethal distending toxin (cdt) genes and the cytotoxic activity in vero and hep-2 cells was estimated in 29 campylobacter jejuni and 36 campylobacter coli from foods, animals and humans isolates. all c. jejuni showed flaa, cadf, cdta, cdtb, cdtc and cdt cluster genes fragments, except for ceue (86.2%) and cdt genes (93.1%). amongst c. coli strains, a lower prevalence of ceue gene (83.3%) was detected than that for cdta, cdtb, cdtc genes (97.2%), cdt gene cluste ...201019195703
inaccuracy of routine susceptibility tests for detection of erythromycin resistance of campylobacter jejuni and campylobacter coli.in the netherlands, both an increase in and regional differences in erythromycin resistance of campylobacter jejuni and campylobacter coli have been reported. to determine the accuracy of routine tests for erythromycin resistance, 48 erythromycin-resistant isolates from various laboratories that participate in the dutch surveillance of campylobacter infections were reinvestigated. initial susceptibility testing for erythromycin had been performed by disk diffusion in six and mic-based methods in ...201019438640
occurrence of salmonella, campylobacter, yersinia enterocolitica, escherichia coli o157 and listeria monocytogenes in swine.the objective of this study was to investigate the occurrence of major bacterial foodborne pathogens in swine. in total, 359 samples from manure storage tanks (91) and fresh pooled faeces (268) obtained from finisher (110), sows (78) and weanlings (80) were collected and tested. campylobacter, salmonella, yersinia enterocolitica, escherichia coli o157 and listeria monocytogenes were isolated from 36.5%, 31.5%, 5.8%, 3.3% and 3.3% of samples respectively. all e. coli o157 isolates found on 10 far ...201019638163
demonstration of persistent strains of campylobacter jejuni within broiler farms over a 1-year period in lithuania.the aim of the study was to investigate the flock prevalence of campylobacter jejuni and campylobacter coli in broiler farms in lithuania and to identify possible persistent strains of camp. jejuni using amplified fragment length polymorphism (aflp) typing method.201019709341
dna identification and characterization of campylobacter jejuni and campylobacter coli isolated from caecal samples of chickens in grenada.to speciate campylobacter strains from the caeca of chickens in grenada using pcr and to evaluate dna-based typing methods for the characterization of these isolates.201019735321
campylobacter jejuni and campylobacter coli genotyping by high-resolution melting analysis of a flaa fragment.the highly variable flagellin-encoding flaa gene has long been used for genotyping campylobacter jejuni and campylobacter coli. high-resolution melting (hrm) analysis is emerging as an efficient and robust method for discriminating dna sequence variants. the objective of this study was to apply hrm analysis to flaa-based genotyping. the initial aim was to identify a suitable flaa fragment. it was found that the pcr primers commonly used to amplify the flaa short variable repeat (svr) yielded a m ...201019933351
species-specific identification and differentiation of arcobacter, helicobacter and campylobacter by full-spectral matrix-associated laser desorption/ionization time of flight mass spectrometry analysis.rapid and reliable identification of arcobacter and helicobacter species, and their distinction from phenotypically similar campylobacter species, has become increasingly important, since many of them are now recognized as human and/or animal pathogens. matrix-associated laser desorption/ionization-time of flight (maldi-tof) ms has been shown to be a rapid and sensitive method for characterization of micro-organisms. in this study, we therefore established a reference database of selected arcoba ...201019959629
diversity of campylobacter jejuni and campylobacter coli genotypes from human and animal sources from rio de janeiro, brazil.to compare the genotypes of campylobacter jejuni and campylobacter coli isolates of human and animal origin collected in rio de janeiro city, 30 c. jejuni and 35 c. coli isolates from animal sources (n=45) and human patients with gastroenteritis (n=20) were genotyped by pcr-based techniques, namely random amplified polymorphic dna (rapd-pcr) and enterobacterial repetitive intergenic consensus sequence (eric-pcr). rapd-pcr identified 50 types and eric-pcr identified 22 genotypes, among the 65 cam ...201019765787
comparison between the biofilm initiation of campylobacter jejuni and campylobacter coli strains to an inert surface using biofilm ring test.the adhesion to an inert surface (the first step of biofilm formation) of the two main pathogenic campylobacter species, campylobacter jejuni and campylobacter coli, isolated from diverse origins, was compared.201019796124
study of cytolethal distending toxin (cdt) in campylobacter coli using a multiplex polymerase chain reaction assay and its distribution among clinical and food strains.from a collection of 242 food isolates of campylobacter spp. 72 were identified as campylobacter jejuni and 170 as campylobacter coli and of 112 isolates from clinical sources, 92 were identified as c. jejuni and 20 as c. coli. the results for food isolates were in contrast to other reported identifications. twenty c. coli strains from clinical cases of campylobacter that occurred in portugal and 170 isolated from poultry products were examined for the presence of cytolethal distending toxin (cd ...201019821742
development and evaluation of internal amplification controls for use in a real-time duplex pcr assay for detection of campylobacter coli and campylobacter jejuni.a common problem of both conventional and real-time pcr assays is failure of dna amplification due to the presence of inhibitory substances in samples. in view of this, our aim was to develop and evaluate internal amplification controls (iacs) for use with an existing duplex real-time pcr assay for campylobacter coli and campylobacter jejuni. both competitive and non-competitive iacs were developed and evaluated. the competitive approach involved a dna fragment of the coding region of the fish v ...201019833779
investigation of food and environmental exposures relating to the epidemiology of campylobacter coli in humans in northwest england.this study uses multilocus sequence typing (mlst) to investigate the epidemiology of campylobacter coli in a continuous study of a population in northwest england. all cases of campylobacter identified in four local authorities (government administrative boundaries) between 2003 and 2006 were identified to species level and then typed, using mlst. epidemiological information was collected for each of these cases, including food and recreational exposure variables, and the epidemiologies of c. je ...201019854914
a genomic island defines subspecies-specific virulence features of the host-adapted pathogen campylobacter fetus subsp. venerealis.the pathogen campylobacter fetus comprises two subspecies, c. fetus subsp. fetus and c. fetus subsp. venerealis. although these taxa are highly related on the genome level, they are adapted to distinct hosts and tissues. c. fetus subsp. fetus infects a diversity of hosts, including humans, and colonizes the gastrointestinal tract. in contrast, c. fetus subsp. venerealis is largely restricted to the bovine genital tract, causing epidemic abortion in these animals. in light of their close genetic ...201019897645
a genomic island defines subspecies-specific virulence features of the host-adapted pathogen campylobacter fetus subsp. venerealis.the pathogen campylobacter fetus comprises two subspecies, c. fetus subsp. fetus and c. fetus subsp. venerealis. although these taxa are highly related on the genome level, they are adapted to distinct hosts and tissues. c. fetus subsp. fetus infects a diversity of hosts, including humans, and colonizes the gastrointestinal tract. in contrast, c. fetus subsp. venerealis is largely restricted to the bovine genital tract, causing epidemic abortion in these animals. in light of their close genetic ...201019897645
development of a dna microarray to detect antimicrobial resistance genes identified in the national center for biotechnology information database.to understand the mechanisms and epidemiology of antimicrobial resistance (ar), the genetic elements responsible must be identified. due to the myriad of possible genes, a high-density genotyping technique is needed for initial screening. to achieve this, ar genes in the national center for biotechnology information genbank database were identified by their annotations and compiled into a nonredundant list of 775 genes. a dna microarray was constructed of 70mer oligonucelotide probes designed to ...201019916789
genetic similarity of campylobacter isolates in humans, food, and water sources in central poland.campylobacter spp. is an important cause of gastroenteritis in humans throughout the world. however, sources of these infections are often difficult to identify. therefore, this study aimed at analyzing the genetic relatedness of campylobacter isolates from environmental and food samples as well as stool specimens of diarrheal patients obtained in a single geographic region in poland. only a few campylobacter jejuni isolates (4/18, 22%) could be assigned to one cluster, whereas the majority were ...201020001328
recrudescent campylobacter jejuni infection in an immunocompetent adult following experimental infection with a well-characterized organism.the recrudescence of infection with campylobacter jejuni after appropriate antibiotic treatment has not been previously reported in an immunocompetent adult. we present the complete clinical, microbiologic, and immunologic evaluation of a closely monitored healthy male with recrudescent c. jejuni infection occurring in the absence of immunodeficiency following experimental infection with a well-characterized strain. after antibiotic treatment, the initial infection was clinically cleared and mic ...201019923572
characterization of two diesel fuel degrading microbial consortia enriched from a non acclimated, complex source of microorganisms.the bioremediation of soils impacted by diesel fuels is very often limited by the lack of indigenous microflora with the required broad substrate specificity. in such cases, the soil inoculation with cultures with the desired catabolic capabilities (bioaugmentation) is an essential option. the use of consortia of microorganisms obtained from rich sources of microbes (e.g., sludges, composts, manure) via enrichment (i.e., serial growth transfers) on the polluting hydrocarbons would provide biorem ...201020158909
prolonged survival of campylobacter species in bovine manure compost.the persistence of naturally occurring campylobacteria in aerobic compost constructed of manure from beef cattle that were administered chlortetracycline and sulfamethazine (as700) or from cattle not administered antibiotics (control) was examined. although there were no differences in population sizes of heterotrophic bacteria, the temperature of as700 compost was more variable and did not become as high as that of control compost. there were significant differences in water content, total carb ...201020023098
comparison of pcr binary typing (p-bit), a new approach to epidemiological subtyping of campylobacter jejuni, with serotyping, pulsed-field gel electrophoresis, and multilocus sequence typing methods.to overcome some of the deficiencies with current molecular typing schema for campylobacter spp., we developed a prototype pcr binary typing (p-bit) approach. we investigated the distribution of 68 gene targets in 58 campylobacter jejuni strains, one campylobacter lari strain, and two campylobacter coli strains for this purpose. gene targets were selected on the basis of distribution in multiple genomes or plasmids, and known or putative status as an epidemicity factor. strains were examined wit ...201020023103
proteomic analysis of iron acquisition, metabolic and regulatory responses of yersinia pestis to iron starvation.the gram-negative bacterium yersinia pestis is the causative agent of the bubonic plague. efficient iron acquisition systems are critical to the ability of y. pestis to infect, spread and grow in mammalian hosts, because iron is sequestered and is considered part of the innate host immune defence against invading pathogens. we used a proteomic approach to determine expression changes of iron uptake systems and intracellular consequences of iron deficiency in the y. pestis strain kim6+ at two phy ...201020113483
activation of human and chicken toll-like receptors by campylobacter spp.campylobacter infection in humans is accompanied by severe inflammation of the intestinal mucosa, in contrast to colonization of chicken. the basis for the differential host response is unknown. toll-like receptors (tlrs) sense and respond to microbes in the body and participate in the induction of an inflammatory response. thus far, the interaction of campylobacter with chicken tlrs has not been studied. here, we investigated the potential of four campylobacter strains to activate human tlr1/2/ ...201020038539
activation of human and chicken toll-like receptors by campylobacter spp.campylobacter infection in humans is accompanied by severe inflammation of the intestinal mucosa, in contrast to colonization of chicken. the basis for the differential host response is unknown. toll-like receptors (tlrs) sense and respond to microbes in the body and participate in the induction of an inflammatory response. thus far, the interaction of campylobacter with chicken tlrs has not been studied. here, we investigated the potential of four campylobacter strains to activate human tlr1/2/ ...201020038539
antimicrobial susceptibilities of multidrug-resistant campylobacter jejuni and c. coli strains: in vitro activities of 20 antimicrobial agents.there is a paucity of information regarding antimicrobial agents that are suitable to treat severe infections caused by multidrug-resistant campylobacter spp. our aim was to identify agents that are potentially effective against multiresistant campylobacter strains. the in vitro activities of 20 antimicrobial agents against 238 campylobacter strains were analyzed by determining mics by the agar plate dilution method or the etest. these strains were selected from 1,808 campylobacter isolates coll ...201020038624
high-throughput identification of bacteria and yeast by matrix-assisted laser desorption ionization-time of flight mass spectrometry in conventional medical microbiology laboratories.matrix-assisted laser desorption ionization-time of flight mass spectrometry (maldi-tof ms) is suitable for high-throughput and rapid diagnostics at low costs and can be considered an alternative for conventional biochemical and molecular identification systems in a conventional microbiological laboratory. first, we evaluated maldi-tof ms using 327 clinical isolates previously cultured from patient materials and identified by conventional techniques (vitek-ii, api, and biochemical tests). discre ...201020053859
amoebae and algae can prolong the survival of campylobacter species in co-culture.several species of free-living amoebae can cause disease in humans. however, in addition to the direct pathogenicity of e.g. acanthamoebae and naegleria species, they are recognized as environmental hosts, indirectly involved in the epidemiology of many pathogenic bacteria. although several studies have demonstrated intracellular survival of many different bacteria in these species, the extent of such interactions as well as the implications for the epidemiology of the bacterial species involved ...201020056117
mass spectrometric analysis of ehrlichia chaffeensis tandem repeat proteins reveals evidence of phosphorylation and absence of glycosylation.ehrlichia chaffeensis has a small subset of immunoreactive secreted, acidic (pi approximately 4), tandem repeat (tr)-containing proteins (trps), which exhibit abnormally large electrophoretic masses that have been associated with glycosylation of the tr domain.201020209062
comparison of campylobacter populations isolated from a free-range broiler flock before and after slaughter.relatively little is known about the campylobacter genotypes colonizing extensively reared broiler flocks and their survival through the slaughter process, despite the increasing demand for free-range and organic products by the consumer. campylobacter isolates from a free-range boiler flock, sampled before and after slaughter, were genotyped by mlst (multilocus sequence typing) and sequence analysis of the flaa short variable region (svr). the campylobacter genotypes isolated before and after s ...201020071049
understanding the molecular epidemiology of the footrot pathogen dichelobacter nodosus to support control and eradication programs.the gram-negative anaerobe dichelobacter nodosus is the primary etiologic agent of ovine footrot. few studies of the genetic diversity and epidemiology of d. nodosus have been done, despite the economic cost and welfare implications of the disease. this study examined a large collection of australian isolates; 735 isolates from footrot-infected sheep from 247 farms in western australia (wa) were tested by pulsed-field gel electrophoresis (pfge), and a subset of 616 isolates was tested by infrequ ...201020071558
understanding the molecular epidemiology of the footrot pathogen dichelobacter nodosus to support control and eradication programs.the gram-negative anaerobe dichelobacter nodosus is the primary etiologic agent of ovine footrot. few studies of the genetic diversity and epidemiology of d. nodosus have been done, despite the economic cost and welfare implications of the disease. this study examined a large collection of australian isolates; 735 isolates from footrot-infected sheep from 247 farms in western australia (wa) were tested by pulsed-field gel electrophoresis (pfge), and a subset of 616 isolates was tested by infrequ ...201020071558
expanded multilocus sequence typing and comparative genomic hybridization of campylobacter coli isolates from multiple hosts.the purpose of this work was to evaluate the evolutionary history of campylobacter coli isolates derived from multiple host sources and to use microarray comparative genomic hybridization to assess whether there are particular genes comprising the dispensable portion of the genome that are more commonly associated with certain host species. genotyping and clonalframe analyses of an expanded 16-gene multilocus sequence typing (mlst) data set involving 85 isolates from 4 different hosts species te ...201020097815
methods for determining disease burden and calibrating national surveillance data in the united kingdom: the second study of infectious intestinal disease in the community (iid2 study).infectious intestinal disease (iid), usually presenting as diarrhoea and vomiting, is frequently preventable. though often mild and self-limiting, its commonness makes iid an important public health problem. in the mid 1990s around 1 in 5 people in england suffered from iid a year, costing around pound0.75 billion. no routine information source describes the uk's current community burden of iid. we present here the methods for a study to determine rates and aetiology of iid in the community, pre ...201020444246
risk assessment of growth hormones and antimicrobial residues in meat.growth promoters including hormonal substances and antibiotics are used legally and illegally in food producing animals for the growth promotion of livestock animals. hormonal substances still under debate in terms of their human health impacts are estradiol-17β, progesterone, testosterone, zeranol, trenbolone, and melengestrol acetate (mga) . many of the risk assessment results of natural steroid hormones have presented negligible impacts when they are used under good veterinary practices. for ...201024278538
diversity and impact of prokaryotic toxins on aquatic environments: a review.microorganisms are ubiquitous in all habitats and are recognized by their metabolic versatility and ability to produce many bioactive compounds, including toxins. some of the most common toxins present in water are produced by several cyanobacterial species. as a result, their blooms create major threats to animal and human health, tourism, recreation and aquaculture. quite a few cyanobacterial toxins have been described, including hepatotoxins, neurotoxins, cytotoxins and dermatotoxins. these t ...201022069558
abc transporters involved in export of cell surface glycoconjugates.complex glycoconjugates play critical roles in the biology of microorganisms. despite the remarkable diversity in glycan structures and the bacteria that produce them, conserved themes are evident in the biosynthesis-export pathways. one of the primary pathways involves representatives of the atp-binding cassette (abc) transporter superfamily. these proteins are responsible for the export of a wide variety of cell surface oligo- and polysaccharides in both gram-positive and gram-negative bacteri ...201020805402
role of the pjm1 plasmid-encoded transport proteins fatb, c and d in ferric anguibactin uptake in the fish pathogen vibrio anguillarum.vibrio anguillarum serotype o1 is part of the natural flora in the aquatic habitat, but under certain circumstances it can cause terminal haemorrhagic septicemia in marine and fresh water fish due to the action of the anguibactin iron uptake system encoded by the virulence plasmid pjm1. this plasmid harbours the genes for the biosynthesis of the siderophore anguibactin and the ferric anguibactin transport proteins fatd, c, b and a encoded in the iron transport operon. the fata protein is the out ...201021304833
typing of campylobacter jejuni and campylobacter coli isolated from live broilers and retail broiler meat by flaa-rflp, mlst, pfge and rep-pcr.we analyzed 100 campylobacter spp. isolates (c. jejuni and c. coli) from grenada, puerto rico and alabama, which were collected from live broilers or retail broiler meat. we analyzed these isolates with four molecular typing methods: restriction fragment length polymorphism of the flaa gene (flaa-rflp), multilocus sequence typing (mlst), pulsed-field gel electrophoresis (pfge), and automated repetitive extragenic palindromic polymerase chain reaction (rep-pcr) using the diversilab system. all me ...201021130125
molecular characterization of the 16s rrna gene of helicobacter fennelliae isolated from stools and blood cultures from paediatric patients in south africa.forty strains of h. fennelliae collected from paediatric blood and stool samples over an 18 year period at a children's hospital in cape town, south africa, were amplified by pcr of the 16s rrna. two distinct genotypes of h. fennelliae were identified based on the phylogenetic analysis. this was confirmed by sequencing a portion of the beta subunit of the rna polymerase (rpob) gene. all isolates from south africa clustered with a proposed novel helicobacter strain (accession number af237612) iso ...201022567323
molecular characterization of the 16s rrna gene of helicobacter fennelliae isolated from stools and blood cultures from paediatric patients in south africa.forty strains of h. fennelliae collected from paediatric blood and stool samples over an 18 year period at a children's hospital in cape town, south africa, were amplified by pcr of the 16s rrna. two distinct genotypes of h. fennelliae were identified based on the phylogenetic analysis. this was confirmed by sequencing a portion of the beta subunit of the rna polymerase (rpob) gene. all isolates from south africa clustered with a proposed novel helicobacter strain (accession number af237612) iso ...201022567323
tir domain-containing adaptor sarm is a late addition to the ongoing microbe-host dialog.toll/interleukin-1 receptor (tir) domain-containing proteins play important roles in defense against pathogens in both animals and plants, connecting the immunity signaling pathways via a chain of specific protein-protein interactions. among them is sarm, the only tir domain-containing adaptor that can negatively regulate tlr signaling. by extensive phylogenetic analysis, we show here that sarm is closely related to bacterial proteins with tir domains, suggesting that this family has a different ...201021110998
simultaneous screening of 24 target genes of foodborne pathogens in 35 foodborne outbreaks using multiplex real-time sybr green pcr analysis.a set of 8 multiplex real-time sybr green pcr (sg-pcr) assays including 3 target primers and an internal amplification control (iac) primer was simultaneously evaluated in 3 h or less with regard to detection of 24 target genes of 23 foodborne pathogens in 7 stool specimens of foodborne outbreak using a 96-well reaction plate. this assay, combined with dna extraction (qiaamp dna stool mini kit), offered detection of greater than 10(3)-10(4) foodborne pathogens per g in stool specimens. the produ ...201020936159
tir domain-containing adaptor sarm is a late addition to the ongoing microbe-host dialog.toll/interleukin-1 receptor (tir) domain-containing proteins play important roles in defense against pathogens in both animals and plants, connecting the immunity signaling pathways via a chain of specific protein-protein interactions. among them is sarm, the only tir domain-containing adaptor that can negatively regulate tlr signaling. by extensive phylogenetic analysis, we show here that sarm is closely related to bacterial proteins with tir domains, suggesting that this family has a different ...201021110998
solexaqa: at-a-glance quality assessment of illumina second-generation sequencing data.illumina's second-generation sequencing platform is playing an increasingly prominent role in modern dna and rna sequencing efforts. however, rapid, simple, standardized and independent measures of run quality are currently lacking, as are tools to process sequences for use in downstream applications based on read-level quality data.201020875133
the era of molecular and other non-culture-based methods in diagnosis of sepsis.sepsis, a leading cause of morbidity and mortality throughout the world, is a clinical syndrome with signs and symptoms relating to an infectious event and the consequent important inflammatory response. from a clinical point of view, sepsis is a continuous process ranging from systemic inflammatory response syndrome (sirs) to multiple-organ-dysfunction syndrome (mods). blood cultures are the current "gold standard" for diagnosis, and they are based on the detection of viable microorganisms pres ...201020065332
flagellin as an adjuvant: cellular mechanisms and potential.flagellin is a potent activator of a broad range of cell types involved in innate and adaptive immunity. an increasing number of studies have demonstrated the effectiveness of flagellin as an adjuvant, as well as its ability to promote cytokine production by a range of innate cell types, trigger a generalized recruitment of t and b lymphocytes to secondary lymphoid sites, and activate tlr5(+)cd11c(+) cells and t lymphocytes in a manner that is distinct from cognate ag recognition. the plasticity ...201021048152
microbial diagnostic microarray for food- and water-borne pathogens.a microbial diagnostic microarray for the detection of the most relevant bacterial food- and water-borne pathogens and indicator organisms was developed and thoroughly validated. the microarray platform based on sequence-specific end labelling of oligonucleotides and the pyhylogenetically robust gyrb marker gene allowed a highly specific (resolution on genus/species level) and sensitive (0.1% relative and 10(4) cfu absolute detection sensitivity) detection of the target pathogens. validation was ...201021255342
successful treatment of cryptosporidiosis in 2 common marmosets (callithrix jacchus) by using paromomycin.two pair-housed, 1-y-old common marmosets (callithrix jacchus) had intermittent loose feces and weight loss for approximately 2 mo. cryptosporidum parvum was identified by elisa in the feces of both animals. cbc and blood chemistry values, including liver enzymes, were within normal range. both marmosets were treated with the antibiotic paromomycin (15 mg/kg po) twice daily for 28 d. resolution of clinical signs coincided with treatment. three follow-up samples, taken 2 wk apart after treatment ...201021205457
multiplex pcr to diagnose bloodstream infections in patients admitted from the emergency department with sepsis.sepsis is caused by a heterogeneous group of infectious etiologies. early diagnosis and the provision of appropriate antimicrobial therapy correlate with positive clinical outcomes. current microbiological techniques are limited in their diagnostic capacities and timeliness. multiplex pcr has the potential to rapidly identify bloodstream infections and fill this diagnostic gap. we identified patients from two large academic hospital emergency departments with suspected sepsis. the results of a m ...200919846634
multiplex pcr to diagnose bloodstream infections in patients admitted from the emergency department with sepsis.sepsis is caused by a heterogeneous group of infectious etiologies. early diagnosis and the provision of appropriate antimicrobial therapy correlate with positive clinical outcomes. current microbiological techniques are limited in their diagnostic capacities and timeliness. multiplex pcr has the potential to rapidly identify bloodstream infections and fill this diagnostic gap. we identified patients from two large academic hospital emergency departments with suspected sepsis. the results of a m ...200919846634
occurrence of campylobacter jejuni and campylobacter coli and their biotypes in beef and dairy cattle from the south of chile.the prevalence of campylobacter jejuni and campylobacter coli and their biotypes in beef and dairy cattle from the south of chile was established. campylobacter were statistically more prevalent among beef cattle (35.9%) than among dairy cattle (21.3%), being c. jejuni the species most frequently isolated.200924031386
rapid identification of bacterial pathogens using a pcr- and microarray-based assay.during the course of a bacterial infection, the rapid identification of the causative agent(s) is necessary for the determination of effective treatment options. we have developed a method based on a modified broad-range pcr and an oligonucleotide microarray for the simultaneous detection and identification of 12 bacterial pathogens at the species level. the broad-range pcr primer mixture was designed using conserved regions of the bacterial topoisomerase genes gyrb and pare. the primer design a ...200919664269
comprehensive and rapid real-time pcr analysis of 21 foodborne outbreaks.a set of four duplex sybr green i pcr (sg-pcr) assay combined with dna extraction using qiaamp dna stool mini kit was evaluated for the detection of foodborne bacteria from 21 foodborne outbreaks. the causative pathogens were detected in almost all cases in 2 hours or less. the first run was for the detection of 8 main foodborne pathogens in 5 stool specimens within 2 hours and the second run was for the detection of other unusual suspect pathogens within a further 45 minutes. after 2 to 4 days, ...200920016673
drug efflux pump deficiency and drug target resistance masking in growing bacteria.recent experiments have shown that drug efflux pump deficiency not only increases the susceptibility of pathogens to antibiotics, but also seems to "mask" the effects of mutations, that decrease the affinities of drugs to their intracellular targets, on the growth rates of drug-exposed bacteria. that is, in the presence of drugs, the growth rates of drug-exposed wt and target mutated strains are the same in a drug efflux pump deficient background, but the mutants grow faster than wt in a drug ef ...200919416855
prevalence of intestinal parasitic and bacterial pathogens in diarrhoeal and non-diarroeal human stools from vhembe district, south africa.in the present study, a cross-sectional survey of intestinal parasitic and bacterial infections in relation to diarrhoea in vhembe district and the antimicrobial susceptibility profiles of isolated bacterial pathogens was conducted. stool samples were collected from 528 patients attending major public hospitals and 295 children attending two public primary schools and were analyzed by standard microbiological and parasitological techniques. entamoeba histolytica/e. dispar (34.2%) and cryptospori ...200920099757
prevalence and risk factor investigation of campylobacter species in beef cattle feces from seven large commercial feedlots in alberta, canada.this fecal prevalence study targeted cattle from 7 large (10,000 to > 40,000 head) commercial feedlots in alberta as a means of establishing campylobacter levels in cattle just prior to animals entering the food chain. overall, 87% [95% confidence interval (ci) = 86-88] of 2776 fresh pen-floor fecal samples were culture positive for campylobacter species, with prevalences ranging from 76% to 95% among the 7 feedlots. campylobacter spp. prevalence was 88% (95% ci = 86-90) in the summer (n = 1376) ...200920046629
prevalence of campylobacter spp. in raw retail poultry on sale in northern ireland.a year-long survey of fresh, retail poultry products on sale in northern ireland was undertaken to define the prevalence of campylobacter spp. by using protocols based on iso (standard) 10272-1:2006. incubation at 37 and 42 degrees c was undertaken to increase the diversity of isolates obtained. overall, 652 isolates were identified as campylobacter spp. by using pcr and amplified fragment length polymorphic typing. phenotyping wrongly identified 21% of isolates. prevalences of campylobacter fou ...200919777882
occurrence of thermophilic campylobacter spp. contamination on vegetable farms in malaysia.the aim of the present study was to examine the prevalence of thermophilic campylobacter spp. (campylobacter jejuni and campylobacter coli) in soil, poultry manure, irrigation water, and freshly harvested vegetables from vegetable farms in malaysia. c. jejuni was detected in 30.4% and 2.7% of the soil samples, 57.1% and 0% of the manure samples, and 18.8% and 3% of the vegetable samples from farm a and farm b, respectively, when using the mpnpcr method. campylobacter spp. was not found in any of ...200919996696
efficacy of mini vidas for the detection of campylobacter spp. from retail broiler meat enriched in bolton broth, with or without the supplementation of blood.the goals of this study were to evaluate the efficacy of the mini vidas automated immunoassay chemistry system to detect campylobacter spp. from retail broiler meat enriched in bolton broth supplemented with lysed blood (b+b) or without blood (b-b), and to detect positive samples at 24 versus 48 h after enrichment. retail broiler meat was enriched and tested for campylobacter spp. with the mini vidas and with an agar plate. isolates were speciated with a multiplex pcr and typed with pulsed-field ...200919903413
campylobacter species occurrence within internal organs and tissues of commercial caged leghorn laying hens.campylobacter spp. are frequently present in the intestinal tract and internal tissues of broiler breeder and broiler chickens. campylobacter spp. ecology in commercial leghorn laying hens has not been extensively studied. the objectives of the current study were to determine 1) campylobacter spp. presence in the reproductive tract, lymphoid organs, liver-gallbladder, and ceca of commercial leghorn laying hens; 2) species of campylobacter present; and 3) antimicrobial resistance pattern of campy ...200919834099
characterization of the oxidative stress stimulon and perr regulon of campylobacter jejuni.during gut colonization, the enteric pathogen campylobacter jejuni must surmount the toxic effects of reactive oxygen species produced by its own metabolism, the host immune system, and intestinal microflora. elucidation of c. jejuni oxidative stress defense mechanisms is critical for understanding campylobacter pathophysiology.200919835633
campylobacter excreted into the environment by animal sources: prevalence, concentration shed, and host association.an intensive study of 443 isolates of campylobacter jejuni and campylobacter coli from 2031 fecal samples excreted by animal sources including cattle, sheep, and pigs, a range of wild and domesticated avian species and pets is described. the prevalence found in the majority of animal sources ranged from 22% to 28% with poultry being highest at 41% and cats and dogs lowest (<5%). the average count excreted for each animal source was found not to be significantly different ranging from approximate ...200919839759
long-term survival of campylobacter jejuni at low temperatures is dependent on polynucleotide phosphorylase activity.campylobacter jejuni is a leading cause of bacterial gastroenteritis worldwide. infection generally occurs after ingestion of contaminated poultry products, usually conserved at low temperatures. the mechanisms promoting survival of c. jejuni in the cold remain poorly understood despite several investigations. the present study provides insight into the survival mechanism by establishing the involvement of polynucleotide phosphorylase (pnpase), a 3'-5' exoribonuclease with multiple biological fu ...200919801468
functional analysis of the helicobacter pylori flagellar switch proteins.helicobacter pylori uses flagellum-mediated chemotaxis to promote infection. bacterial flagella change rotational direction by changing the state of the flagellar motor via a subcomplex referred to as the switch. intriguingly, the h. pylori genome encodes four switch complex proteins, flim, flin, fliy, and flig, instead of the more typical three of escherichia coli or bacillus subtilis. our goal was to examine whether and how all four switch proteins participate in flagellation. previous work de ...200919767432
genetic diversity and stability of the pora allele as a genetic marker in human campylobacter infection.the major outer-membrane protein (momp) of campylobacter jejuni and campylobacter coli, encoded by the pora gene, is extremely genetically diverse. conformational momp epitopes are important in host immunity, and variation in surface-exposed regions probably occurs as a result of positive immune selection during infection. pora diversity has been exploited in genotyping studies using highly discriminatory nucleotide sequences to identify potentially epidemiologically linked cases of human campyl ...200919744989
cost analysis and antimicrobial susceptibility testing comparing the e test and the agar dilution method in campylobacter jejuni and campylobacter coli.although numerous reports have compared the antimicrobial susceptibility of campylobacter spp., controversy still exists about the use of the e test as an alternative to the agar dilution method suggested by the clinical and laboratory standards institute. mics of 8 antimicrobials were determined using the e test and agar dilution methods for 103 campylobacter jejuni and campylobacter coli isolates from fresh chicken randomly purchased from stores in 3 southern ontario counties. overall, 72.6% o ...200919748428
functional characterization of flagellin glycosylation in campylobacter jejuni 81-176.the major flagellin of campylobacter jejuni strain 81-176, flaa, has been shown to be glycosylated at 19 serine or threonine sites, and this glycosylation is required for flagellar filament formation. some enzymatic components of the glycosylation machinery of c. jejuni 81-176 are localized to the poles of the cell in an flhf-independent manner. flagellin glycosylation could be detected in flagellar mutants at multiple levels of the regulatory hierarchy, indicating that glycosylation occurs inde ...200919749047
mouse intestine colonization ability of campylobacter coli strains.campylobacter coli is an etiological agent of gastrointestinal and extraintestinal infections in man and animals, and can be found as a commensal in gastrointestinal tract of animals. in this study, we aimed to determine differences among c coli strains in colonization of the intestinal tract of mice. seven c coli strains isolated from diarrheic patients, asymptomatic hosts and chicken carcasses were used for this study. each strain was inoculated with 0.1 ml of a bacterial suspension (3 x 10(8) ...200919753795
molecular, antigenic, and functional characteristics of ferric enterobactin receptor cfra in campylobacter jejuni.the ferric enterobactin receptor cfra not only is responsible for high-affinity iron acquisition in campylobacter jejuni but also is essential for c. jejuni colonization in animal intestines. in this study, we determined the feasibility of targeting the iron-regulated outer membrane protein cfra for immune protection against campylobacter colonization. alignment of complete cfra sequences from 15 campylobacter isolates showed that the levels of amino acid identity for cfra range from 89% to 98%. ...200919737895
description and sources of contamination by campylobacter spp. of river water destined for human consumption in brittany, france.presence or absence of campylobacter spp. in water of five rivers upstream from an intake point for drinking water production was investigated, and isolates genetically compared with human, pig and poultry isolates in order to determine their source. river water and drinking water obtained from these rivers were sampled one time per month, over a period of one year, and tested for campylobacter. isolates were typed by pfge. campylobacter was not detected in treated drinking water, but 50% of the ...200919942377
thermotolerant coliforms are not a good surrogate for campylobacter spp. in environmental water.this study aimed to assess the importance of quantitatively detecting campylobacter spp. in environmental surface water. the prevalence and the quantity of campylobacter spp., thermotolerant coliforms, and escherichia coli in 2,471 samples collected weekly, over a 2-year period, from 13 rivers and 12 streams in the eastern townships, québec, canada, were determined. overall, 1,071 (43%), 1,481 (60%), and 1,463 (59%) samples were positive for campylobacter spp., thermotolerant coliforms, and e. c ...200919734335
novel clonal complexes with an unknown animal reservoir dominate campylobacter jejuni isolates from river water in new zealand.campylobacter jejuni is widely distributed in the environment, and river water has been shown to carry high levels of the organism. in this study, 244 c. jejuni isolates from three river catchment areas in new zealand were characterized using multilocus sequence typing. forty-nine of the 88 sequence types identified were new. the most common sequence types identified were st-2381 (30 isolates), st-45 (25 isolates), and st-1225 (23 isolates). the majority of the sequence types identified in the r ...200919648367
susceptibility of campylobacter jejuni and campylobacter coli isolated from animals and humans to ciprofloxacin.fifty five thermophilic campylobacter spp. strains were isolated from the caecum of broilers, cecum and colon of pigs and from human faeces. the strains were identified as campylobacter jejuni and campylobacter coli. the more prevalent species in humans and broilers was campylobacter jejuni, and in pigs campylobacter coli. in the framework of this study, sensitivity to ciprofloxacin in isolated strains of campylobacter jejuni and campylobacter coli was tested by e-test. resistant to ciprofloxaci ...200919645360
the response of campylobacter jejuni to low temperature differs from that of escherichia coli.human infection with campylobacter jejuni is often associated with the consumption of foods that have been exposed to both chilling and high temperatures. despite the public health importance of this pathogen, little is known about the effects of cold exposure on its ability to survive a subsequent heat challenge. this work examined the effect of rapid exposure to chilling, as would occur in poultry processing, on the heat resistance at 56 degrees c of two c. jejuni strains, 11168 and 2097e48, a ...200919648365
amoxicillin therapy of poultry flocks: effect upon the selection of amoxicillin-resistant commensal campylobacter spp.the aim of this study was to investigate the effect of amoxicillin therapy of poultry flocks upon the persistence of commensal campylobacter spp. and the incidence of antibiotic resistance.200919656785
a campylobacter jejuni znua orthologue is essential for growth in low-zinc environments and chick colonization.campylobacter jejuni infection is a leading cause of bacterial gastroenteritis in the united states and is acquired primarily through the ingestion of contaminated poultry products. here, we describe the c. jejuni orthologue of znua in other gram-negative bacteria. znua (cj0143c) is the periplasmic component of a putative zinc abc transport system and is encoded on a zinc-dependent operon with cj0142c and cj0141c, which encode the other two likely components of the transport system of c. jejuni. ...200919103921
a campylobacter jejuni znua orthologue is essential for growth in low-zinc environments and chick colonization.campylobacter jejuni infection is a leading cause of bacterial gastroenteritis in the united states and is acquired primarily through the ingestion of contaminated poultry products. here, we describe the c. jejuni orthologue of znua in other gram-negative bacteria. znua (cj0143c) is the periplasmic component of a putative zinc abc transport system and is encoded on a zinc-dependent operon with cj0142c and cj0141c, which encode the other two likely components of the transport system of c. jejuni. ...200919103921
antimicrobial activity of prulifloxacin tested against a worldwide collection of gastroenteritis-producing pathogens, including those causing traveler's diarrhea.prulifloxacin, the prodrug of ulifloxacin (active component), is a newer fluoroquinolone with broad activity against enteric and nonenteric gram-negative bacilli. ulifloxacin and other oral comparator agents were tested for activity against 582 gastroenteritis strains from global surveillance studies. ulifloxacin was highly active against escherichia coli, salmonella spp., shigella spp., yersinia spp., vibrio spp., aeromonas spp., and plesiomonas spp. (mic(50)s and mic(90)s, <or=0.03 microg/ml a ...200919114678
antimicrobial activity of prulifloxacin tested against a worldwide collection of gastroenteritis-producing pathogens, including those causing traveler's diarrhea.prulifloxacin, the prodrug of ulifloxacin (active component), is a newer fluoroquinolone with broad activity against enteric and nonenteric gram-negative bacilli. ulifloxacin and other oral comparator agents were tested for activity against 582 gastroenteritis strains from global surveillance studies. ulifloxacin was highly active against escherichia coli, salmonella spp., shigella spp., yersinia spp., vibrio spp., aeromonas spp., and plesiomonas spp. (mic(50)s and mic(90)s, <or=0.03 microg/ml a ...200919114678
diversity and habitat specificity of free-living protozoa in commercial poultry houses.despite stringent biosecurity measures, infections by bacterial food pathogens such as campylobacter are a recurrent problem in industrial poultry houses. as the main transmission route remains unclear, persistence of these infections has been linked to bacterial survival and possibly multiplication within protozoan vectors. to date, however, virtually no information is available on the diversity and occurrence of free-living protozoa in these environments. using a combination of microscopic ana ...200919124593
[determination of sensitivity to antibiotics of campylobacter jejuni and campylobacter coli isolated from human feces].one of the most important bacterial zoonosis is campylobacteriosis. human disease is mostly caused by thermophilic campylobacter spp: campylobacter jejuni (c. jejuni), campylobacter coli (c. coli), campylobacter lari (c. lari) and campylobacter upsakensis (campsalensis). campylobacteriosis is a mild and self-healing disorder. in patients with more severe and prolonged forms, an antibiotic treatment is recommended. recommended drugsare erythromycin, ciprofloxacin, tetracyclin, chloramphenicol and ...200919195264
novel campylobacter lari-like bacteria from humans and molluscs: description of campylobacter peloridis sp. nov., campylobacter lari subsp. concheus subsp. nov. and campylobacter lari subsp. lari subsp. nov.a polyphasic study was undertaken to clarify the taxonomic position of campylobacter lari-like strains isolated from shellfish and humans. the diversity within the strain collection was initially screened by means of fluorescent amplified fragment length polymorphism analysis and whole-cell protein electrophoresis, revealing the existence of two clusters distinct from c. lari and other campylobacter species. the divergence of these clusters was confirmed by phenotypic analysis and by 16s rrna an ...200919406805
spatiotemporal homogeneity of campylobacter subtypes from cattle and sheep across northeastern and southwestern scotland.source attribution using molecular subtypes has implicated cattle and sheep as sources of human campylobacter infection. whether the campylobacter subtypes associated with cattle and sheep vary spatiotemporally remains poorly known, especially at national levels. here we describe spatiotemporal patterns of prevalence, bacterial enumeration, and subtype composition in campylobacter isolates from cattle and sheep feces from northeastern (63 farms, 414 samples) and southwestern (71 farms, 449 sampl ...200919700557
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