We evaluated the prevalence and epidemiology of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli isolates in pigs during production cycle on a Czech farm with the history of previous use of ceftiofur. ESBL-producing E. coli isolates were obtained from rectal swabs from pigs of different age groups (suckling piglets, weaned piglets, growers and sows). Collected samples were directly cultivated on MacConkey agar with cefotaxime (2 mg l-1 ), whereas intestinal swabs of slaughtered pigs and surface swabs from pig carcasses were also pre-enriched in buffered peptone water without antimicrobials before the cultivation. Clonal relationship of selected isolates was determined by XbaI pulse-field gel electrophoresis and multi-locus sequence typing. The transferability of plasmids carrying blaCTX-M genes was tested by conjugation experiments. From all examined samples, 141 (43·7%, n = 323) were positive for ESBL-producing E. coli. All ESBL-producing isolates showed resistance to multiple antimicrobials and were positive for blaCTX-M genes. The blaCTX-M-1 was carried by conjugative IncN/ST1 plasmids (c. 40-45 kb) while the blaCTX-M-15 was located on conjugative F plasmids with F:18:A5:B1 formula (c. 165 kb). This study demonstrated the persistence of CTX-M-positive E. coli isolates 2 months after banner of ceftiofur usage and indicated possible risk of transmission of these isolates to humans via the food chain.
- MeSH
- beta-laktamasy genetika metabolismus MeSH
- Escherichia coli enzymologie genetika izolace a purifikace MeSH
- farmy MeSH
- infekce vyvolané Escherichia coli mikrobiologie patofyziologie veterinární MeSH
- lidé MeSH
- multilokusová sekvenční typizace MeSH
- nemoci prasat mikrobiologie patofyziologie MeSH
- plazmidy genetika metabolismus MeSH
- prasata růst a vývoj mikrobiologie MeSH
- proteiny z Escherichia coli genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
The spread of antimicrobial resistance from human activity derived sources to natural habitats implicates wildlife as potential vectors of antimicrobial resistance transfer. Wild birds, including corvid species can disseminate mobile genetic resistance determinants through faeces. This study aimed to determine the occurrence of plasmid-mediated quinolone resistance (PMQR) genes in Escherichia coli and Klebsiella spp. isolates obtained from winter roosting sites of American crows (Corvus brachyrhynchos) and common ravens (Corvus corax) in Canada. Faecal swabs were collected at five roosting sites across Canada. Selective media isolation and multiplex PCR screening was utilized to identify PMQR genes followed by gene sequencing, pulse-field gel electrophoresis and multilocus sequence typing to characterize isolates. Despite the low prevalence of E. coli containing PMQR (1·3%, 6/449), qnrS1, qnrB19, qnrC, oqxAB and aac(6')-Ib-cr genes were found in five sequence types (ST), including E. coli ST 131. Conversely, one isolate of Klebsiella pneumoniae contained the plasmid-mediated resistance gene qnrB19. Five different K. pneumoniae STs were identified, including two novel types. The occurrence of PMQR genes and STs of public health significance in E. coli and Klebsiella pneumoniae recovered from corvids gives further evidence of the anthropogenic derived dissemination of antimicrobial resistance determinants at the human activity-wildlife-environment interface. SIGNIFICANCE AND IMPACT OF THE STUDY: This study examined large corvids as possible vector species for the dissemination of antimicrobial resistance in indicator and pathogenic bacteria as a means to assess the anthropogenic dissemination of plasmid-mediated quinolone resistance (PMQR) genes. Although rare, PMQR genes were found among corvid populations across Canada. The clinically important Escherichia coli strain ST131 containing aac(6')-Ib-cr gene along with a four-class phenotypic antimicrobial resistance (AMR) pattern as well as one Klebsiella pneumoniae strain containing a qnrB19 gene were identified in one geographical location. Corvids are a viable vector for the circulation of PMQR genes and clinically important clones in wide-ranging environments.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence genetika MeSH
- chinolony farmakologie MeSH
- Escherichia coli účinky léků genetika izolace a purifikace MeSH
- infekce bakteriemi rodu Klebsiella farmakoterapie mikrobiologie MeSH
- infekce vyvolané Escherichia coli farmakoterapie mikrobiologie MeSH
- Klebsiella pneumoniae účinky léků genetika izolace a purifikace MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- multilokusová sekvenční typizace MeSH
- nemoci ptáků farmakoterapie mikrobiologie MeSH
- plazmidy genetika MeSH
- průřezové studie MeSH
- vrány mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Kanada MeSH
- Publikační typ
- abstrakt z konference MeSH
Current diagnostic techniques are inefficient in distinguishing latent and low-risk forms of prostate cancer from high-risk forms. The present study is focused on determination of putative tumor markers of aggressive high-grade forms of prostate cancer. Potential markers were determined in blood sera of 133 patients (82 cases and 51 controls) and in cell lines (Gleason score 9-derived 22Rv1 and normal tissue derived PNT1A) on mRNA and protein levels. Alpha-methylacyl-CoA racemase (AMACR), metallothionein classes 1A and 2A (MT1A and MT2A) were determined and compared to prostate specific antigen (PSA) levels. On mRNA level, significantly increased expression of MT2A (2.4-fold), PSA (2.6-fold) and AMACR (8.4-fold) and insignificantly (1.9-fold) elevated MT1A in 22Rv1 compared to non-tumor PNT1A were determined. On protein level, significant enhancement of free PSA and total PSA in tumor cell line was evident. AMACR protein was 1.5-fold elevated in tumor line (below the level of significance). Contrary to mRNA, significantly (p = 0.01) reduced level of MT protein in tumor lines was determined. In the case of serum level, significantly enhanced MT level (4.5-fold) in patients' sera was found. No significant changes were observed in the case of AMACR. These findings indicate possible alternative role of MT to PSA prostate cancer marker. In addition, level of AMACR is distinctly higher in the Gleason score 9 in serum of patients and MT shows a descending trend in relation to Gleason score.
- MeSH
- adenokarcinom genetika metabolismus patologie MeSH
- dospělí MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé středního věku MeSH
- lidé MeSH
- messenger RNA genetika MeSH
- metalothionein genetika metabolismus MeSH
- nádorové biomarkery genetika metabolismus MeSH
- nádory prostaty genetika metabolismus patologie MeSH
- prognóza MeSH
- prostatický specifický antigen genetika metabolismus MeSH
- racemasy a epimerasy genetika metabolismus MeSH
- senioři MeSH
- studie případů a kontrol MeSH
- stupeň nádoru MeSH
- western blotting MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Současné možnosti diagnostiky karcinomu prostaty neumožňují odlišit agresivní (signifikantní tumory) od klinicky latentních (nesignifikantních forem). Práce se zaměřuje na stanovení hladiny klinicky využitelných tumorových markerů karcinomu prostaty: alfa-metyl CoA-racemáza (AMACR), caveolin-1, metalothionein (MT), p53, NF-?B, c-FOS, c-JUN, Ki-67, prostatický specifický antigen (PSA), ZIP1 a ZnT-1 na úrovni prostatické tkáně (reprezentované buněčnými liniemi 22Rv1 a PNT1A) a v séru pacientů s histologicky ověřeným adenokarcinomem (82 karcinomových a 51 kontrolních sér). Na úrovni mRNA bylo zjištěno snížení exprese Cav-1, NF-?B, c-FOS a c-JUN a zvýšení exprese metalothioneinu, AMACR, PSA, Ki-67, MMP-9 a zinkových transportérů ZIP1 a ZnT-1. V séru je popsáno signifikantní zvýšení metalothioneinu nad hladinu 1 µM. Hladina caveolinu-1 se významně zvyšuje u high-grade tumorů, což jej staví do pozice možného markeru agresivní formy karcinomu prostaty.
Current methods for diagnosis of prostate carcinoma do not enable us to distinguish aggressive tumours (significant tumours) from clinically latent tumours (non-significant tumours). This study aims to determine levels of potential clinically important tumour markers such as alpha-methyl CoA-racemase (AMACR), caveolin-1, metallothionein (MT), p53, NF-?B, c-FOS, c-JUN, Ki-67, prostate-specific antigen (PSA), ZIP1 and ZnT-1 in prostatic tissue represented by 22Rv1 (tumour) and PNT1A (healthy) cell lines and in blood serum of patients with histologically evaluated adenocarcinoma (82 tumour patients and 51 healthy volunteers). Based on mRNA expression, it was found that Cav-1, NF-?B, c-FOS and c-JUN were down-regulated and MT, AMACR, PSA, Ki-67, MMP-9 and zinc transporters ZIP1 and ZnT-1 were up-regulated. In serum, the level of MT was significantly enhanced above 1 µM. Caveolin-1 levels were significantly increased in high-grade tumours, which points to the possibility of using this protein as a marker for the aggressive form of prostate carcinoma.
- Klíčová slova
- nádorová onemocnění, imunodetekce, molekulárně-biologické techniky,
- MeSH
- 2D gelová elektroforéza využití MeSH
- diagnostické techniky molekulární metody MeSH
- elektrochemie metody MeSH
- financování organizované MeSH
- kaveolin 1 diagnostické užití MeSH
- lidé MeSH
- nádorové biomarkery krev MeSH
- nádorové buněčné linie chemie MeSH
- nádory prostaty genetika krev MeSH
- polymerázová řetězová reakce využití MeSH
- prostatický specifický antigen izolace a purifikace krev MeSH
- senzitivita a specificita MeSH
- stupeň závažnosti nemoci MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Klíčová slova
- monitorování buněk v reálném čase, XCELLigence, MTT,
- MeSH
- buněčný cyklus účinky léků MeSH
- doxorubicin farmakologie MeSH
- elipticiny farmakologie MeSH
- financování organizované MeSH
- nádorové buněčné linie cytologie účinky léků MeSH
- neuroblastom farmakoterapie MeSH
- techniky in vitro MeSH
- testy toxicity metody využití MeSH
- MeSH
- apoptóza genetika MeSH
- buněčné linie MeSH
- exprese genu genetika MeSH
- financování organizované MeSH
- geny p53 genetika MeSH
- geny ras genetika MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory prostaty genetika MeSH
- proliferace buněk MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- srovnávací studie MeSH
