Paenibacillus larvae and Melissococcus plutonius represent the most threatening bacterial diseases of honeybee (Apis mellifera)-American and European foulbrood, respectively. For efficient control of those diseases, rapid and accurate detection of the pathogens is crucial. Therefore, we developed a novel multiplex PCR method simultaneously detecting both pathogens. To design and optimize multiplex PCR reaction, four strains of P. larvae representing four ERIC genotypes I-IV (strain DSM 7030-ERIC I, DSM 25430-ERIC II, LMG 16252-ERIC III, DSM 3615-ERIC IV) were selected. Those strains were fully sequenced using long-read sequencing (Sequel I, Pacific Biosciences). For P. larvae, the multicopy insertion sequence IS256 identified in all genotypes of P. larvae was selected to provide high sensitivity. M. plutonius was detected by plasmid pMP1 sequence and the virulence verified by following detection of ETX/MTX2 toxin responsible for pore formation in the cell membrane. As an internal control, a gene encoding for major royal jelly protein 1 specific for honeybees was selected. The method was validated on 36 clinical specimens collected from the colonies suffering from American and European foulbrood in the Czech Republic. Based on the results, sensitivity of PCR was calculated to 93.75% and specificity to 100% for P. larvae diagnosed from hive debris and 100% sensitivity and specificity for honeybee workers and larval scales as well as for diseased brood infected by M. plutonius.
- MeSH
- Enterococcaceae * MeSH
- larva mikrobiologie MeSH
- multiplexová polymerázová řetězová reakce metody MeSH
- Paenibacillus larvae * genetika MeSH
- Paenibacillus * genetika MeSH
- plazmidy genetika MeSH
- transpozibilní elementy DNA MeSH
- včely genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Enterobacterales produkující karbapenemázu (CPE – Carbapenemase-Producing Enterobacterales) představují globální zdravotnický problém. V Evropě dochází zejména v jižních a východních státech ke zvyšování podílu enterobakterií rezistentních vůči karbapenemům. Od roku 2021 roste také v České republice výskyt kmenů s produkcí karbapenemázy, změnila se i struktura zastoupení jednotlivých enzymů, zejména ve prospěch metalobetalaktamázy typu NDM. Tato situace je zřejmě následkem epidemie covid-19 a zhoršenou epidemickou situací ve zdravotnických zařízeních.
Carbapenemase-producing Enterobacterales (CPE) are a global health problem. In Europe, the proportion of carbapenem resistant enterobacteria is increasing, particularly in the southern and eastern countries. Since 2021, there has also been a widespread increase in the occurrence of CPE in the Czech Republic, the structure of carbapenemases has also shifted towards higher proportion of the NDM type. This situation is apparently the result of the COVID-19 epidemic and the worsened epidemic situation in healthcare facilities.
- MeSH
- antibiotická rezistence * genetika imunologie účinky léků MeSH
- COVID-19 MeSH
- mikrobiologie * trendy MeSH
- Publikační typ
- novinové články MeSH
- rozhovory MeSH
Enterobacterales produkující karbapenemázu (CPE – Carbapenemase-Producing Enterobacterales) představují globální zdravotnický problém. V Evropě dochází zejména v jižních státech ke zvyšování podílu enterobakterií rezistentních vůči karbapenemům. Od roku 2016 dochází také v České republice k plošnému nárůstu počtu případů, v roce 2019 bylo zaznamenáno 270 případů kolonizace nebo infekce CPE. Tento trend je zčásti způsoben několika epidemickými epizodami, odráží ale i zhoršující se endemickou situaci ve zdravotnických zařízeních. Přesto lze Českou republiku stále hodnotit jako stát s nízkým výskytem CPE.
Carbapenemase-producing Enterobacterales (CPE) are a global health problem. In Europe, the proportion of carbapenem-resistant enterobacteria is increasing, particularly in the southern states. Since 2016, there has also been a widespread increase in the number of cases in the Czech Republic, with 270 cases of colonization or CPE infection reported in 2019. This trend is partly due to several outbreaks, but it also reflects the deteriorating endemic situation in healthcare facilities. Nevertheless, the Czech Republic still remains a low CPE incidence country.
Antimicrobial consumption in veterinary medicine has led to the spread of multi drug-resistance in clinically important bacteria, with the companion animals and their environment involved as emerging reservoirs. While CTX-M-15 and CMY-2 acquired β-lactamases have been widely detected in the bacterial population of companion and breeding animals in European area, DHA-1 enzymes have been rarely reported in veterinary medicine. The aim of the study was to characterize the Escherichia coli associated with mortality of a litter of Bulldog puppies in a breeding kennel located in Pesaro area, Central Italy. The E. coli strains O39 serotype were resistant to 3rd/4th generation cephalosporins, chloramphenicol, aminoglycosides, trimethoprim-sulfamethoxazole, and ciprofloxacin, retaining susceptibility to carbapenems, colistin, fosfomycin, and levofloxacin (by Microscan Autoscan4, EUCAST clinical breakpoints). Pulse field gel electrophoreses (PFGE-XbaI) on five E. coli strains revealed the presence of a single profile. Whole genome sequencing (WGS) analysis revealed a complex resistome, harboring blaTEM-1b, blaCTX-M-15, blaOXA-1, aph(6)-Ib, aac(6')Ib-cr, aac(3)-Ila, aph(6)-Id, aadA1, qnrB1, sul2, catA1, catB3, tetA, and dfrA14 genes located on a 302597 bp IncHI2/HI2A plasmid. Moreover, blaDHA-1, qnrB4, mph(A), sul1, and dfrA17 determinants were carried on an 83,429 bp IncFII plasmid. A blaCMY-2 determinant was carried on a 90,249 bp IncI1 plasmid. Two IncX1 and IncX4 plasmids without antimicrobial resistance genes were also detected. The presence of lpfA, iss, astA, and gad virulence factors was highlighted. This is the first report in Italy on an invasive infection in eight 2-weeks old dogs caused by the same MDR E. coli O39 blaCTX-M-15, blaCMY-2, blaDHA-1, and aac(6')-Ib-cr positive strain. The above MDR E. coli clone caused the death of the entire litter, despite amoxicillin-clavulanate and enrofloxacin administration. The tank for storage of the water used to prepare the milk-based meal for the litter was the suspected reservoir.
- MeSH
- bakteriální léková rezistence MeSH
- Escherichia coli genetika patogenita MeSH
- fatální výsledek MeSH
- infekce vyvolané Escherichia coli * mikrobiologie veterinární MeSH
- nemoci psů * mikrobiologie MeSH
- plazmidy genetika MeSH
- psi MeSH
- sekvenování celého genomu metody MeSH
- zvířata MeSH
- Check Tag
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
Dermatofyty jsou mikroskopické vláknité houby způsobující časté infekce kůže a kožních adnex. Jejich identifikace byla do nedávné doby prováděna pouze podle morfologických charakteristik. Ta je však velmi zdlouhavá a vyžaduje značné zkušenosti. V posledních letech se v identifikaci dermatofytů začíná uplatňovat MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) hmotnostní spektrometrie (MS). U vláknitých hub je nutné, kvůli jejich komplexní morfologii a požadavkům na kultivaci, použít odlišné postupy zpracování kultury. Tato práce se zabývá srovnáním MALDI-TOF MS v identifikaci dermatofytů v porovnání s metodou sekvenace oblasti ITS (Internal Transcribed Spacer) rDNA a konvenčními metodami. Do studie bylo zahrnuto 81 izolátů dermatofytů získaných z klinických vzorků. Výsledky MALDI-TOF MS analýzy se shodovaly s výsledky molekulárně genetické identifikace u 75 % izolátů. Metodami klasické identifikace se podařilo identifikovat pouze 47 % izolátů. MALDI-TOF MS lze tedy považovat za účinný nástroj rychlé a jednoduché identifikace dermatofytů v rutinní diagnostické laboratoři.
Dermatophytes are microscopic fungi causing skin infections and infections of skin adnexa. Their identification is usually performed by morphological characteristics requiring experienced microbiologists and quiet long turnaround time. In recent years, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been introducing to many diagnostic laboratories allowing identification of dermatophytes as well. For that purpose, however, specific procedures comparing with bacteria and yeasts must be used because of their complex morphology and specific cultivation requirements. This study is focused on comparison of MALDI-TOF MS-based identification, identification using DNA sequencing of Internal Transcribed Spacer (ITS) rDNA region and conventional methods. In the study, 81 clinical isolates of dermatophytes were included. MALDI-TOF MS correctly identified 74% of all isolates. Using classical methods, only 47% of isolates were identified. Thus, MALDI-TOF MS provides a useful tool for identification of those fungi in routine microbiological laboratories.
