Civilization factors are responsible for the increasing of human exposure to mycobacteria from environment, water, and food during the last few decades. Urbanization, lifestyle changes and new technologies in the animal and plant industry are involved in frequent contact of people with mycobacteria. Type 1 diabetes is a multifactorial polygenic disease; its origin is conditioned by the mutual interaction of genetic and other factors. The environmental factors and certain pathogenetic pathways are shared by some immune mediated chronic inflammatory and autoimmune diseases, which are associated with triggers originating mainly from Mycobacterium avium subspecies paratuberculosis, an intestinal pathogen which persists in the environment. Type 1 diabetes and some other chronic inflammatory diseases thus pose the global health problem which could be mitigated by measures aimed to decrease the human exposure to this neglected zoonotic mycobacterium.
Mycobacterium avium subsp. paratuberculosis (MAP) is a pathogenic bacterium causing the paratuberculosis, chronic and infectious disease common particularly in wild and domestic ruminants. Currently, culture techniques to detect viable MAP are still used most commonly, although these require a long incubation period. Consequently, a faster molecular method for assessing MAP cell viability based on cell membrane integrity was introduced consisting of sample treatment with the intercalation dye propidium monoazide (PMA) followed by quantitative PCR (qPCR). However, the PMA-qPCR assay is complicated by demanding procedures involving work in a darkroom and on ice. In this study, we therefore optimized a viability assay combining sample treatment with palladium (Pd) compounds as an alternative viability marker to PMA, which does not require such laborious procedures, with subsequent qPCR. The optimized Pd-qPCR conditions consisting of 90 min exposure to 30 μM bis(benzonitrile)dichloropalladium(II) or 30 μM palladium(II)acetate at 5 °C and using ultrapure water as a resuspension medium resulted in differences in quantification cycle (Cq) values between treated live and dead MAP cells of 8.5 and 7.9, respectively, corresponding to approximately 2.5 log units. In addition, Pd-qPCR proved to be superior to PMA-qPCR in distinguishing between live and dead MAP cells. The Pd-qPCR viability assay thus has the potential to replace time-consuming culture methods and demanding PMA-qPCR in the detection and quantification of viable MAP cells with possible application in food, feed, clinical and environmental samples.
- MeSH
- azidy farmakologie MeSH
- biotest MeSH
- kvantitativní polymerázová řetězová reakce metody MeSH
- mikrobiální viabilita MeSH
- Mycobacterium avium subsp. paratuberculosis * genetika MeSH
- palladium farmakologie MeSH
- paratuberkulóza * mikrobiologie MeSH
- propidium farmakologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Quantitative PCR (qPCR) has become a frequently employed direct method for the detection and quantification of Mycobacterium avium subsp. paratuberculosis (MAP). The quantity of MAP determined by qPCR, however, may be affected by the type of qPCR quantification standard used (PCR product, plasmid, genomic DNA) and the way in which standard DNA quantity is determined (absorbance, fluorescence). In practice, this can be reflected in the inability to properly compare quantitative data from the same qPCR assays in different laboratories. Thus, the aim of this study was to prepare a prototype of an international MAP reference standard, which could be used to calibrate routinely used qPCR quantification standards in various laboratories to promote clinical data comparability. Considering stability, storage and shipment issues, a lyophilised fecal suspension artificially contaminated with a MAP reference strain was chosen as the most suitable form of the standard. The effect of five types of lyophilisation matrices on standard stability was monitored on 2-weeks interval basis for 4 months by F57 qPCR. The lyophilisation matrix with 10% skimmed milk provided the best recovery and stability in time and was thus selected for subsequent comparative testing of the standard involving six diagnostic and research laboratories, where DNA isolation and qPCR assay procedures were performed with the parallel use of the identical supplied genomic DNA solution. Furthermore, the effect of storage conditions on the standard stability was tested for at least 6 months. The storage at room temperature in the dark and under light, at + 4 °C, - 20 °C and - 80 °C showed no significant changes in the stability, and also no substantial changes in MAP viability were found using phage amplification assay. The prepared MAP quantification standard provided homogeneous and reproducible results demonstrating its suitability for utilisation as an international reference qPCR standard.
- MeSH
- DNA bakterií klasifikace genetika MeSH
- feces chemie mikrobiologie MeSH
- kvantitativní polymerázová řetězová reakce normy MeSH
- lyofilizace MeSH
- Mycobacterium avium subsp. paratuberculosis klasifikace genetika izolace a purifikace MeSH
- nemoci skotu diagnóza MeSH
- paratuberkulóza diagnóza mikrobiologie MeSH
- referenční standardy MeSH
- senzitivita a specificita MeSH
- skot MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Tato práce prezentuje komplexní diagnosticko-terapeutický proces u pacientky bez známého imunodeficitu s těžkou generalizovanou infekcí způsobenou Mycobacterium avium complex, projevující se pancytopenií, hepatopatií a renálním selháním. Ojedinělým byl také výskyt pouze mikroskopického nálezu (granulomatózně nekrotizující proces v kostní dřeni a slezině) při absenci makroskopických či radiologických nálezů. Finální diagnóza byla sta-novena kultivačním vyšetřením tkáně sleziny. Po nasazení cílené antituberkulotické léčby došlo ke zlepšení stavu pacientky, vymizení všech příznaků a návratu do běžného života.
This case study describes the complex diagnostic and therapeutic process in an immunocompetent patient suffering from a disseminated nontuberculous mycobacteria infection caused by Mycobacterium avium complex. During the infection, the patient suffered from pancytopenia, liver damage and acute renal failure. Necrotizing granulomatosis was found in bone marrow and splenic tissue with no signs of infection macroscopically or using imaging methods. Diagnosis was eventually determined by microbial cultivation of splenic tissue. The patient was then prescribed targeted antituberculotic treatment, which led to clinical and laboratory improvement.
- MeSH
- antituberkulotika terapeutické užití MeSH
- diagnostické zobrazování metody MeSH
- ethambutol terapeutické užití MeSH
- klarithromycin terapeutické užití MeSH
- klinické laboratorní techniky metody MeSH
- kombinovaná farmakoterapie MeSH
- lidé středního věku MeSH
- lidé MeSH
- Mycobacterium avium izolace a purifikace účinky léků MeSH
- mykobakteriózy * diagnóza patologie terapie MeSH
- pancytopenie MeSH
- rifampin terapeutické užití MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Based on the isosterism concept, we have designed and synthesized homologous N-alkyl-2-[4-(trifluoromethyl)benzoyl]hydrazine-1-carboxamides (from C1 to C18) as potential antimicrobial agents and enzyme inhibitors. They were obtained from 4-(trifluoromethyl)benzohydrazide by three synthetic approaches and characterized by spectral methods. The derivatives were screened for their inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) via Ellman's method. All the hydrazinecarboxamides revealed a moderate inhibition of both AChE and BuChE, with IC50 values of 27.04-106.75 µM and 58.01-277.48 µM, respectively. Some compounds exhibited lower IC50 for AChE than the clinically used drug rivastigmine. N-Tridecyl/pentadecyl-2-[4-(trifluoromethyl)benzoyl]hydrazine-1-carboxamides were identified as the most potent and selective inhibitors of AChE. For inhibition of BuChE, alkyl chain lengths from C5 to C7 are optimal substituents. Based on molecular docking study, the compounds may work as non-covalent inhibitors that are placed in a close proximity to the active site triad. The compounds were evaluated against Mycobacterium tuberculosis H37Rv and nontuberculous mycobacteria (M. avium, M. kansasii). Reflecting these results, we prepared additional analogues of the most active carboxamide (n-hexyl derivative 2f). N-Hexyl-5-[4-(trifluoromethyl)phenyl]-1,3,4-oxadiazol-2-amine (4) exhibited the lowest minimum inhibitory concentrations within this study (MIC ≥ 62.5 µM), however, this activity is mild. All the compounds avoided cytostatic properties on two eukaryotic cell lines (HepG2, MonoMac6).
- MeSH
- acetylcholinesterasa metabolismus MeSH
- antiinfekční látky * chemická syntéza chemie farmakologie MeSH
- buňky Hep G2 MeSH
- butyrylcholinesterasa metabolismus MeSH
- cholinesterasové inhibitory * chemická syntéza chemie farmakologie MeSH
- GPI-vázané proteiny metabolismus MeSH
- imidazoly * chemická syntéza chemie farmakologie MeSH
- lidé MeSH
- Mycobacterium avium růst a vývoj MeSH
- Mycobacterium kansasii růst a vývoj MeSH
- Mycobacterium tuberculosis růst a vývoj MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Paratuberculosis, a chronic disease affecting ruminant livestock, is caused by Mycobacterium avium subsp. paratuberculosis (MAP). It has direct and indirect economic costs, impacts animal welfare and arouses public health concerns. In a survey of 48 countries we found paratuberculosis to be very common in livestock. In about half the countries more than 20% of herds and flocks were infected with MAP. Most countries had large ruminant populations (millions), several types of farmed ruminants, multiple husbandry systems and tens of thousands of individual farms, creating challenges for disease control. In addition, numerous species of free-living wildlife were infected. Paratuberculosis was notifiable in most countries, but formal control programs were present in only 22 countries. Generally, these were the more highly developed countries with advanced veterinary services. Of the countries without a formal control program for paratuberculosis, 76% were in South and Central America, Asia and Africa while 20% were in Europe. Control programs were justified most commonly on animal health grounds, but protecting market access and public health were other factors. Prevalence reduction was the major objective in most countries, but Norway and Sweden aimed to eradicate the disease, so surveillance and response were their major objectives. Government funding was involved in about two thirds of countries, but operations tended to be funded by farmers and their organizations and not by government alone. The majority of countries (60%) had voluntary control programs. Generally, programs were supported by incentives for joining, financial compensation and/or penalties for non-participation. Performance indicators, structure, leadership, practices and tools used in control programs are also presented. Securing funding for long-term control activities was a widespread problem. Control programs were reported to be successful in 16 (73%) of the 22 countries. Recommendations are made for future control programs, including a primary goal of establishing an international code for paratuberculosis, leading to universal acknowledgment of the principles and methods of control in relation to endemic and transboundary disease. An holistic approach across all ruminant livestock industries and long-term commitment is required for control of paratuberculosis.
- MeSH
- chov zvířat MeSH
- divoká zvířata mikrobiologie MeSH
- hlášení nemocí normy MeSH
- incidence MeSH
- Mycobacterium avium subsp. paratuberculosis izolace a purifikace MeSH
- paratuberkulóza ekonomika epidemiologie prevence a kontrola MeSH
- přežvýkavci mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- MeSH
- lidé MeSH
- Mycobacterium avium MeSH
- mykobakteriózy * patofyziologie terapie MeSH
- nemoci střev mikrobiologie patologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- komentáře MeSH
- úvodníky MeSH
Netuberkulózní mykobakteriózy jsou vzácná onemocnění, jejichž incidence spolu s narůstajícím počtem imunokompromitovaných nemocných a lepšími diagnostickými možnostmi pozvolna narůstá. Článek stručně shrnuje problematiku netuberkulózních mykobakterióz na podkladě reálné kazuistiky.
Nontuberculous mycobacterial infections are rare diseases. However, as number of immunocompromised patients is growing and modern diagnostic tools are available, both the importance and incidence of nontuberculous mycobacterial infections are gaining clinical importance. Based on a clinical case, this article briefly summarizes the current knowledge on this issue.
- MeSH
- antibakteriální látky terapeutické užití MeSH
- imunosupresivní léčba škodlivé účinky MeSH
- klinické laboratorní techniky metody MeSH
- lidé MeSH
- Mycobacterium avium MeSH
- mykobakteriózy * diagnóza terapie MeSH
- senioři MeSH
- smrt MeSH
- výsledek terapie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- Publikační typ
- kazuistiky MeSH
Mycobacterium avium subsp. paratuberculosis (MAP), the etiological agent of paratuberculosis, is considered to be a potential zoonotic pathogen and meat is one of the sources of MAP exposure for humans. MAP has been shown to be relatively resistant to different food processing methods, but there is a lack of information about the effects of ripening and fermentation processes on MAP survival in meat. Our results demonstrate that a short ripening process during teewurst production did not reduce MAP counts and viable mycobacteria were detected even during 4 weeks of storage. Although no viable MAP was recovered during the dry fermented sausage production process, there was no reduction in MAP count detected by real time PCR during production and storage of both sausages. Although the impact of foodborne exposure to viable MAP and/or mycobacterial components has not yet been clearly determined, the consumption of raw fermented meat products may be considered as a possible route of MAP transmission to humans.
- MeSH
- DNA bakterií izolace a purifikace MeSH
- fermentace MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- manipulace s potravinami metody MeSH
- masné výrobky mikrobiologie MeSH
- Mycobacterium avium subsp. paratuberculosis genetika růst a vývoj izolace a purifikace MeSH
- paratuberkulóza mikrobiologie MeSH
- prasata MeSH
- skladování potravin MeSH
- skot MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Granulomatózní plicní proces má celou řadu příčin, mezi něž patří infekční, nádorová onemocnění, reakce na organické prachy a chemikálie. Granulom vzniká také u některých imunitních aberací (sarkoidóza), ale i u jiných stavů. Autoři předkládají raritní pozorování, kdy u pacientky byla zjištěna dvojí, odlišná příčina granulomů. Jednalo se o koincidenci parazitárního postižení plic a aviární mykobakteriózy.
Granulomatous pulmonary disease has many possible causes, including infectious and cancer illnesses, reactions to organic dust and chemicals. Granuloma can also occur in some immune aberrations (sarcoidosis) and other conditions. The authors present a rare observation of a patient diagnosed with two different causes of granulomas. It was a coincidence of parasitic lung disease and avian mycobacteriosis.
- MeSH
- granulom * diagnostické zobrazování diagnóza etiologie MeSH
- helmintóza diagnostické zobrazování diagnóza MeSH
- Langhansovy obrovské buňky cytologie MeSH
- lidé MeSH
- Mycobacterium avium MeSH
- parazitární onemocnění plic diagnostické zobrazování diagnóza MeSH
- senioři MeSH
- Check Tag
- lidé MeSH
- senioři MeSH
- ženské pohlaví MeSH
