Anthelmintic resistance (AR) is a serious threat to animal health and has a major economic impact worldwide due to production and financial losses. The aim of this study was to determine the occurrence of AR on 30 goat farms in Slovakia during the pasturing seasons and to compare three widely used in vitro and in vivo methods for detecting AR in field conditions. A three-year survey was conducted during the pasturing seasons of 2014-2016. Goats on each farm were split into treated and control groups and were treated by recommended (5 mg/kg body weight) and double doses (10 mg/kg b.w.) of albendazole. Comparisons between percent reduction in a faecal egg count reduction test (FECRT) and an egg hatch test (EHT) and the presence of L3 larvae in a larval development test (LDT) using resistant concentrations of benzimidazole (BZ) were monitored after treatment. The FECRT indicated percent reductions of 69.2-86.2% for the single dose and of 36.3-45.4% for the double dose. The EHT indicated that all farms had BZ-resistant nematodes. Low (<15% hatching) and high (>15% hatching) levels of resistance were detected on 13 and 17 farms, respectively. The LDT failed to detect resistant larvae on seven farms but detected low and high levels of resistance on seven and 14 farms, respectively. The data indicate a moderate correlation between in vitro and in vivo tests for detecting BZ resistance among the 30 goat farms. The hatching detected by the EHT and the presence of L3 larvae by the LDT at resistant BZ concentrations provided reasonable identification of low levels of resistance in the parasite populations, but the use of a double dose for a treatment may underestimate the real occurrence of low levels of resistant parasites on goat farms.

• Klíčová slova
• Anthelmintics, Benzimidazole, Detection method, Goats, Resistance,
• MeSH
• anthelmintika farmakologie   MeSH
• benzimidazoly farmakologie   MeSH
• Haemonchus anatomie a histologie   účinky léků   růst a vývoj   MeSH
• kozy parazitologie   MeSH
• larva anatomie a histologie   účinky léků   růst a vývoj   MeSH
• léková rezistence * MeSH
• Ostertagia anatomie a histologie   účinky léků   růst a vývoj   MeSH
• techniky in vitro MeSH
• Trichostrongylus anatomie a histologie   účinky léků   růst a vývoj   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Slovenská republika MeSH
• Názvy látek
• anthelmintika MeSH
• benzimidazole MeSH Prohlížeč
• benzimidazoly MeSH

Antibiotic resistance is an ever-increasing global problem. Major commercial antibiotics often fail to fight common bacteria, and some pathogens have become multi-resistant. Polymyxins are potent bactericidal antibiotics against gram-negative bacteria. Known resistance to polymyxin includes intrinsic, mutational and adaptive mechanisms, with the recently described horizontally acquired resistance mechanisms. In this review, we present several strategies for bacteria to develop enhanced resistance to polymyxins, focusing on changes in the outer membrane, efflux and other resistance determinants. Better understanding of the genes involved in polymyxin resistance may pave the way for the development of new and effective antimicrobial agents. We also report novel in silico tested primers for PCR assay that may be able distinguish colistin-resistant isolates carrying the plasmid-encoded mcr genes and will assist in combating the spread of colistin resistance in bacteria.

• Klíčová slova
• LPS, colistin, polymyxin, resistance,
• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální léková rezistence účinky léků   genetika   MeSH
• bakteriální proteiny účinky léků   genetika   MeSH
• gramnegativní bakteriální infekce farmakoterapie   genetika   MeSH
• kolistin farmakologie   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• polymyxiny farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• antibakteriální látky MeSH
• bakteriální proteiny MeSH
• kolistin MeSH
• polymyxiny MeSH

BACKGROUND: In 1928, the first antibiotic, penicillin, was discovered. That was the beginning of a great era in the development and prescription of antibiotics. However, the introduction of these antimicrobial agents into clinical practice was accompanied by the problem of antibiotic resistance. Currently, bacterial resistance to antibiotics poses a major problem in both hospital and community settings throughout the world. METHODS AND RESULTS: This review provides examples of modern genetic methods and their practical application in the field of extended-spectrum β-lactamase detection. Since extended-spectrum β-lactamases are the main mechanism of Gram-negative bacterial resistance to oxyimino-cephalosporins, rapid and accurate detection is requested in common clinical practice. CONCLUSIONS: Currently, the detection of extended-spectrum β-lactamases is primarily based on the determination of bacterial phenotypes rather than genotypes. This is because therapeutic decisions are based on assessing the susceptibility rather than presence of resistance genes. One of the main disadvantages of genetic methods is high costs, including those of laboratory equipment. On the other hand, if these modern methods are introduced into diagnostics, they often help in rapid and accurate detection of certain microorganisms or their resistance and pathogenic determinants.

• MeSH
• beta-laktamová rezistence * genetika   MeSH
• genetické techniky * ekonomika   normy   MeSH
• molekulární biologie metody   MeSH
• náklady a analýza nákladů MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Clostridioides difficile is one of the most important human pathogens. The identification of its possible sources is important for the understanding of C. difficile infection (CDI) epidemiology. A total of 16 water samples from wastewater and surface water in South Moravia in the Czech Republic and 82 samples of fish and gulls were collected between May and July 2019. C. difficile isolates were cultured by direct plating and after enrichment on chromogenic media. Susceptibility testing to eight antimicrobials was performed by Etest. C. difficile isolates were characterized by ribotyping, multilocus sequence typing, multilocus tandem repeats analysis, and toxin gene detection. Samples from fish and gulls were C. difficile negative; a total of 15 C. difficile isolates from 8 out of 16 water samples were cultured (6 out of 14 surface water samples yielded 6 isolates, and 2 out of 2 wastewater samples yielded 9 isolates). Direct plating was culture positive in 6 out of 16 samples (12 isolates), and enrichment culture was positive in an additional 2 out of 16 samples (3 isolates). Twelve different ribotyping profiles and 14 sequence types of clades 1, 4, and 5 were identified. Five isolates did not carry genes for toxins, and eight isolates carried genes for toxins A and B; the remaining two isolates (RT078) carried the genes for toxins A, B, and binary. All C. difficile isolates were susceptible to amoxicillin, moxifloxacin, tetracycline, and vancomycin and resistant to ciprofloxacin. A high level of erythromycin resistance (>256 mg/L) was detected in eight isolates. Clindamycin resistance was found in 14 isolates, 6 of which showed a high level of resistance (>256 mg/L) and carried ermB. Surprisingly, one isolate (RT010, ST15) showed resistance to metronidazole (12 mg/L) with the presence of the plasmid pCD-METRO. In conclusion, a diverse spectrum of C. difficile strains was found in wastewater and surface water. A recently discovered plasmid-bound resistance to metronidazole was detected in C. difficile from the surface water sample. IMPORTANCE The combination of direct plating and culture after enrichment was used in order to gain a spectrum of C. difficile ribotypes present in the water samples. Toxigenic C. difficile ribotypes detected in surface water and in wastewater treatment plants overlapped with those derived from patients with CDI and/or animals. Importantly, a recently discovered plasmid-mediated resistance to metronidazole, a drug used for the treatment of CDI, was detected in C. difficile from river water.

• Klíčová slova
• MLST, antimicrobial resistance, erm(B), plasmid-bound metronidazole resistance, ribotyping, surface water, wastewater treatment plant,
• MeSH
• antibakteriální látky farmakologie   terapeutické užití   MeSH
• bakteriální léková rezistence genetika   MeSH
• Clostridioides difficile * genetika   MeSH
• Clostridioides MeSH
• klostridiové infekce * veterinární   MeSH
• lidé MeSH
• metronidazol farmakologie   terapeutické užití   MeSH
• mikrobiální testy citlivosti MeSH
• odpadní voda MeSH
• plazmidy genetika   MeSH
• řeky MeSH
• voda MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antibakteriální látky MeSH
• metronidazol MeSH
• odpadní voda MeSH
• voda MeSH

There is a steady increase in the number of overweight and obese people worldwide and increasingly, younger people. Excess adipose tissue impairs the action of insulin, leading to insulin resistance (IR). Tissue IR is a major factor in relation to cardiovascular disease, metabolic syndrome and diabetes. Thus, it is important to recognize at the pre-disease stage with the possibility of therapeutic intervention. IR is assessed using indicators of epidemiological significance, most often calculated from fasting and postprandial glucose and insulin values, so-called indirect indicators of insulin resistance. The most commonly used parameter is the Homeostatic Model Assessment (HOMA). Although the Quantitative Insulin Sensitivity Check Index (QUICKI), Matsuda Index and the Insulin Secretion-Sensitivity Index-2 (ISSI-2) are also used, the values of these indices established for IR vary for different age, sex, populations and ethnic groups. Thus, appropriate reference values of indirect indices should be determined for such groups, and when this is precluded, data from published studies carried out on the most ethnically, socio-economically and age-matched populations should be applied.

• Klíčová slova
• decision limit, insulin resistance, reference interval,
• MeSH
• glukózový toleranční test metody   MeSH
• glykemický index MeSH
• homeostáza fyziologie   MeSH
• inzulinová rezistence fyziologie   MeSH
• krevní glukóza metabolismus   MeSH
• lidé MeSH
• referenční hodnoty MeSH
• sekrece inzulinu fyziologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• krevní glukóza MeSH

Staphylococcal hospital isolates (n = 166) were tested in a touchdown multiplex-polymerase chain reaction assay for the identification of methicillin and mupirocin resistance and discrimination of S. aureus (femA gene) from coagulase negative staphylococci and other bacteria. All isolates harbored the 16SrDNA (Staphylococcus genus specific internal control) gene, and 130 (78 %) the mecA (methicillin resistance) gene. Fifty-seven (44 %) of these were determined as methicillin-resistant S. aureus, while the remaining 73 (56 %) were methicillin-resistant coagulase-negative staphylococci. Seventy-five (45 %) isolates harbored the ileS-2 (high-level mupirocin resistance) gene and were determined as mupirocin-resistant. This assay represents a simple, rapid, reliable approach for the detection and discrimination of methicillin-and mupirocin-resistant staphylococci.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• DNA bakterií genetika   MeSH
• lidé MeSH
• methicilin farmakologie   MeSH
• mnohočetná bakteriální léková rezistence * MeSH
• mupirocin farmakologie   MeSH
• nemocnice MeSH
• polymerázová řetězová reakce metody   MeSH
• rezistence na methicilin MeSH
• RNA ribozomální 16S genetika   MeSH
• senzitivita a specificita MeSH
• stafylokokové infekce mikrobiologie   MeSH
• Staphylococcus účinky léků   genetika   izolace a purifikace   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• Názvy látek
• antibakteriální látky MeSH
• bakteriální proteiny MeSH
• DNA bakterií MeSH
• methicilin MeSH
• mupirocin MeSH
• RNA ribozomální 16S MeSH

In the last two decades, microbiology laboratories have radically changed by the introduction of novel technologies, like Next-Generation Sequencing (NGS) and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS). Nevertheless, emergence of antibiotic-resistant microorganisms represents a global threat of current medicine, being responsible for increasing mortality and health-care direct and indirect costs. In addition, the identification of antibiotic-resistant microorganisms, like OXA-48 carbapenemase-producing Enterobacteriaceae, has been changeling for clinical microbiology laboratories. Even the cost of NGS technology and MALDI-TOF MS equipment is relatively high, both technologies are increasingly used in diagnostic and research protocols. Therefore, the aim of this review is to present applications of these technologies used in clinical microbiology, especially in detection of antibiotic resistance and its surveillance, and to propose a combinatory approach of MALDI-TOF MS and NGS for the investigation of microbial associated infections.

• Klíčová slova
• MLST, NGS, beta-lactamase, carbapenemase, susceptibility testing,
• MeSH
• antibakteriální látky farmakologie   MeSH
• Bacteria klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• bakteriální infekce diagnóza   mikrobiologie   MeSH
• bakteriální léková rezistence * MeSH
• hmotnostní spektrometrie metody   MeSH
• laboratoře nemocniční MeSH
• lidé MeSH
• vysoce účinné nukleotidové sekvenování metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• antibakteriální látky MeSH

An occurrence of resistance to tetracycline (TET) and erythromycin (ERY) was ascertained in 82 isolates of Enterococcus spp. of animal and environmental origin. Using E test, 33 isolates were resistant to TET and three isolates to ERY. Using polymerase chain reaction (PCR; single and multiplex), the TET determinants tet(M) and tet(L) were detected in 35 and 13 isolates, respectively. Twelve isolates carried both tet(M) and tet(L) genes. Eight isolates possessed ermB gene associated with ERY resistance. Multiplex PCR was shown to be a suitable method for simultaneous determination of all three resistance determinants that occurred most frequently in bacteria isolated from poultry. This study also demonstrates that gastrointestinal tract of broilers may be a reservoir of enterococci with acquired resistance to both TET and ERY that can be transferred to humans via food chain.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální geny účinky léků   MeSH
• DNA bakterií analýza   MeSH
• drůbež mikrobiologie   MeSH
• Enterococcus účinky léků   genetika   izolace a purifikace   MeSH
• gastrointestinální trakt mikrobiologie   MeSH
• makrolidy farmakologie   MeSH
• mikrobiální testy citlivosti MeSH
• mnohočetná bakteriální léková rezistence genetika   MeSH
• polymerázová řetězová reakce MeSH
• tetracyklin farmakologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antibakteriální látky MeSH
• DNA bakterií MeSH
• makrolidy MeSH
• tetracyklin MeSH

Common alimentary obesity frequently occurs on a polygenic basis as a typical lifestyle disorder in the developed countries. It is associated with characteristic complex metabolic changes, which are the cornerstones for future metabolic syndrome development. The aims of our study were 1) to determine the incidence of metabolic syndrome (based on the diagnostic criteria defined by the International Diabetes Federation for children and adolescents) in Czech obese children, 2) to evaluate the incidence of insulin resistance according to HOMA-IR and QUICKI homeostatic indexes in obese children with and without metabolic syndrome, and 3) to consider the diagnostic value of these indexes for the early detection of metabolic syndrome in obese children. We therefore performed anthropometric and laboratory examinations to determine the incidence of metabolic syndrome and insulin resistance in the group of 274 children with obesity (128 boys and 146 girls) aged 9-17 years. Metabolic syndrome was found in 102 subjects (37 %). On the other hand, the presence of insulin resistance according to QUICKI <0.357 was identified in 86 % and according to HOMA-IR >3.16 in 53 % of obese subjects. This HOMA-IR limit was exceeded by 70 % children in the MS(+) group, but only by 43 % children in the MS(-) group (p<0.0001). However, a relatively high incidence of insulin resistance in obese children without metabolic syndrome raises a question whether the existing diagnostic criteria do not falsely exclude some cases of metabolic syndrome. On the basis of our results we suggest to pay a preventive attention also to obese children with insulin resistance even if they do not fulfill the actual diagnostic criteria for metabolic syndrome.

• MeSH
• časná diagnóza MeSH
• dítě MeSH
• incidence MeSH
• inzulin krev   MeSH
• inzulinová rezistence * MeSH
• kauzalita MeSH
• komorbidita MeSH
• krevní glukóza analýza   MeSH
• lidé MeSH
• metabolický syndrom krev   diagnóza   epidemiologie   MeSH
• mladiství MeSH
• obezita krev   diagnóza   epidemiologie   MeSH
• reprodukovatelnost výsledků MeSH
• rizikové faktory MeSH
• senzitivita a specificita MeSH
• surveillance populace MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Názvy látek
• inzulin MeSH
• krevní glukóza MeSH

The occurrence of colistin resistance has increased rapidly among Enterobacterales around the world. We performed a national survey of plasmid-mediated colistin resistance in human clinical isolates through a retrospective analysis of samples from 2009 to 2017 and a prospective sampling in 2018-2020. The aim of this study was to identify and characterize isolates with mcr genes from various regions of the Czech Republic using whole genome sequencing (WGS). Of all 1932 colistin-resistant isolates analyzed, 73 (3.8%) were positive for mcr genes. Most isolates carried mcr-1 (48/73) and were identified as Escherichia coli (n = 44) and Klebsiella pneumoniae (n = 4) of various sequence types (ST). Twenty-five isolates, including Enterobacter spp. (n = 24) and Citrobacter freundii (n = 1) carrying the mcr-9 gene were detected; three of them (Enterobacter kobei ST54) co-harbored the mcr-4 and mcr-9 genes. Multi-drug resistance phenotype was a common feature of mcr isolates and 14% (10/73) isolates also co-harbored clinically important beta-lactamases, including two isolates with carbapenemases KPC-2 and OXA-48. Phylogenetic analysis of E. coli ST744, the dominant genotype in this study, with the global collection showed Czech isolates belonged to two major clades, one containing isolates from Europe, while the second composed of isolates from diverse geographical areas. The mcr-1 gene was carried by IncX4 (34/73, 47%), IncHI2/ST4 (6/73, 8%) and IncI2 (8/73, 11%) plasmid groups. Small plasmids belonging to the ColE10 group were associated with mcr-4 in three isolates, while mcr-9 was carried by IncHI2/ST1 plasmids (4/73, 5%) or the chromosome (18/73, 25%). We showed an overall low level of occurrence of mcr genes in colistin-resistant bacteria from human clinical samples in the Czech Republic.

• Klíčová slova
• Enterobacterales, antibiotic resistance, human, mcr, plasmids,
• Publikační typ
• časopisecké články MeSH