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Версія від 10:25, 15 лютого 2017, створена Curve2pocket (обговореннявнесок) (Створена сторінка: The aim of this study is to determine the presence and diversity of C.?difficile in Dutch animals and to compare the isolates for genetic relatedness to those f...)

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The aim of this study is to determine the presence and diversity of C.?difficile in Dutch animals and to compare the isolates for genetic relatedness to those from patients hospitalized in the Netherlands by means Alectinib cell line of MLVA. Faecal samples were collected from healthy poultry, pigs, veal calves and dairy cattle (100 samples each) at abattoirs during 2009 and 2010 by the Dutch Food and Consumer Product Safety Authority (monitoring samples). One faecal specimen per epidemiological unit (herd or flock) was obtained from arbitrarily selected, apparently healthy animals representing the Dutch animal populations. Samples were stored in buffered peptone water with 10% glycerol (w/v) at ?80��C. Additionally, faecal specimens submitted for routine microbiological diagnostic procedures (virology, bacteriology and/or parasitology) from diarrheic animals were tested (diagnostic samples). These were collected arbitrarily during 2009 and 2010 and stored without preservatives at ?20��C. Samples from dogs (n?=?116), cats (n?=?115), horses (n?=?135), poultry (n?=?21), sheep (n?=?11) and dairy cattle (n?=?5) were obtained from the Veterinary Microbiological Diagnostic Centre of the Faculty of Veterinary Medicine in Utrecht (VMDC); pig samples (n?=?36) were collected from the PI3K inhibition Animal Health Service in Deventer (AHS). Human isolates were collected at the National Reference Laboratory for C.?difficile at Leiden University Medical Centre (LUMC) from patients with diarrhoea who tested positive for C.?difficile toxin or from a surveillance study for CDI in hospitalized patients in 19 hospitals. The prevalence of human PCR ribotypes was based on 1552 samples collected from January 2009 to August 2010. MLVA data involved randomly selected PCR ribotype 012, ribotype 014 and ribotype 078 isolates from January 2006 to August 2010. The culture method involved heat shock treatment (60�� Tryptophan synthase in water at 60��C), after which samples were inoculated onto selective media, Clostridium difficile Selective Medium (Oxoid PB5054A; Oxoid, Basingstoke, UK) and Brazier��s Clostridium difficile selective agar (Oxoid PB5191A). Plates were incubated anaerobically in jars (Mart; Anoxomat, Lichtenvoorde, the Netherlands) for 7?days at 37��C. Enrichment of heat shock-treated samples was performed during 7?days (1?g in 9?mL BHI broth supplemented with cycloserine-cefoxitin (Clostridium difficile Selective Supplement SR0096E; Oxoid, Basingstoke, UK) and 0.1% sodium taurocholate), followed by subculturing on selective agar media. At regular intervals, plates were examined for suspect colonies (morphology, typical odour and positive latex slide agglutination test (Oxoid)), which were pure cultured on Heart Infusion Sheep blood agar (HIS) and stored in buffered peptone water with 20% glycerol (w/v) at ?80��C.