Ticks and the pathogens they transmit constitute a growing burden for human and animal health worldwide. Traditionally, tick-borne pathogen detection has been carried out using PCR-based methods that rely in known sequences for specific primers design. This approach matches with the view of a 'single-pathogen' epidemiology. Recent results, however, have stressed the importance of coinfections in pathogen ecology and evolution with impact in pathogen transmission and disease severity. New approaches, including high-throughput technologies, were then used to detect multiple pathogens, but they all need a priori information on the pathogens to search. Thus, those approaches are biased, limited and conceal the complexity of pathogen ecology. Currently, next generation sequencing (NGS) is applied to tick-borne pathogen detection as well as to study the interactions between pathogenic and non-pathogenic microorganisms associated to ticks, the pathobiome. The use of NGS technologies have surfaced two major points: (i) ticks are associated to complex microbial communities and (ii) the relation between pathogens and microbiota is bidirectional. Notably, a new challenge emerges from NGS experiments, data analysis. Discovering associations among a high number of microorganisms is not trivial and therefore most current NGS studies report lists of microorganisms without further insights. An alternative to this is the combination of NGS with analytical tools such as network analysis to unravel the structure of microbial communities associated to ticks in different ecosystems.
- MeSH
- Bacteria isolation & purification MeSH
- Host-Pathogen Interactions MeSH
- Ticks microbiology MeSH
- Coinfection microbiology MeSH
- Humans MeSH
- Microbial Interactions MeSH
- Microbiota * MeSH
- Tick-Borne Diseases diagnosis microbiology MeSH
- High-Throughput Nucleotide Sequencing MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Review MeSH
The aim of the review is to characterize Enterobacter sakazakii alias Cronobacter sp. as one of most dangerous microbial contaminants in foods. The microbe is a pathogen causing a serious disease in premature and newborn infants, rarely in adults. Despite a low occurrence of the infections, a high mortality (up to 80 %) has been reported. A survey of methods of its detection is given. Cultivation, chromogenic/fluorogenic media, biochemical kits and molecular genetic methods are promising in Enterobacter sakazakii screening in foods.
- MeSH
- Cronobacter sakazakii pathogenicity MeSH
- Molecular Diagnostic Techniques methods MeSH
- Financing, Organized MeSH
- Food Contamination legislation & jurisprudence MeSH
- Culture Techniques methods MeSH
- Polymerase Chain Reaction methods MeSH
- Food Microbiology legislation & jurisprudence MeSH
- Reagent Kits, Diagnostic utilization MeSH
Bakteriální zoonózy představují i v současnosti závažný medicínský problém. K méně častým, ale závažným zoonózám patří maltská horečka způsobená bakterií Brucella melitensis. Předkládaná kazuistika popisuje případ relapsu importované maltské horečky u mladého muže. Diagnóza byla vedle čtyř sérologických testů verifikována přímým průkazem původce v hemokultuře. Izolát Brucella melitensis byl identifikován metodou polymerázové řetězové reakce a metodou hmotností spektrometrie MALDI-TOF.
Bacterial zoonoses still represent a serious medical problem. One of the less frequent but severe zoonoses is brucellosis caused by the bacterium Brucella melitensis. The presented case report describes relapsing imported brucellosis in a young male. In addition to four serological tests, the diagnosis was confirmed by direct detection of the pathogen in blood culture. The isolate of Brucella melitensis was identified using the MALDI-TOF BioTyper method and subsequently also by PCR.
- Keywords
- MALDI-TOF, alimentární infekce, zoonóza,
- MeSH
- Brucella melitensis isolation & purification MeSH
- Brucellosis diagnosis microbiology MeSH
- Travel MeSH
- Humans MeSH
- Young Adult MeSH
- Check Tag
- Humans MeSH
- Young Adult MeSH
- Male MeSH
- Publication type
- Case Reports MeSH
Aphanomyces astaci, the causal agent of the crayfish plague, has recently been confirmed to infect also freshwater-inhabiting crabs. We experimentally tested the resistance of freshwater shrimps, another important decapod group inhabiting freshwaters, to this pathogen. We exposed individuals of two Asian shrimp species, Macrobrachium dayanum and Neocaridina davidi, to zoospores of the pathogen strain isolated from Procambarus clarkii, a known A. astaci carrier likely to get into contact with shrimps. The shrimps were kept in separate vessels up to seven weeks; exuviae and randomly chosen individuals were sampled throughout the experiment. Shrimp bodies and exuviae were tested for A. astaci presence by a species-specific quantitative PCR. The results were compared with amounts of A. astaci DNA in an inert substrate to distinguish potential pathogen growth in live specimens from persisting spores or environmental DNA attached to their surface. In contrast to susceptible crayfish Astacus astacus, we did not observe mortality of shrimps. The amount of detected pathogen DNA was decreasing steadily in the inert substrate, but it was still detectable several weeks after zoospore addition, which should be considered in studies relying on molecular detection of A. astaci. Probably due to moulting, the amount of A. astaci DNA was decreasing in N. davidi even faster than in the inert substrate. In contrast, high pathogen DNA levels were detected in some non-moulting individuals of M. dayanum, suggesting that A. astaci growth may be possible in tissues of this species. Further experiments are needed to test for the potential of long-term A. astaci persistence in freshwater shrimp populations.
Surface plasmon resonance (SPR) biosensors capable of in real time detection of Cronobacter at concentrations down to 10⁶ cells mL⁻¹ in samples of consumer fresh-whole fat milk, powder whole-fat milk preparation, and powder infant formulation were developed for the first time. Antibodies against Cronobacter were covalently attached onto polymer brushes of poly(2-hydroxyethyl methacrylate) (poly(HEMA)) grafted from the SPR chip surface. The lowest detection limit, 10⁴ cells mL⁻¹, was achieved in phosphate buffered saline (pH 7.4) with sensors prepared by covalent immobilization of the same antibodies onto a self assembled monolayer (SAM) of hexa(ethylene glycol) undecanethiol (EG₆). However, when the EG₆ based sensors were challenged with milk samples the non-specific response due to the deposition of non-targeted compounds from the milk samples was much higher than the specific response to Cronobacter hampering the detection in milk. Similar interfering fouling was observed on antifouling polymer brushes of hydroxy-capped oligoethylene glycol methacrylate and even a 10 times higher fouling was observed on the widely used SAM of mixed hydroxy- and carboxy-terminated alkanethiols. Only poly(HEMA) brushes totally suppressed the fouling from milk samples. The robust well-controlled surface initiated atom transfer radical polymerization of HEMA allowed the preparation of highly dense brushes with a minimal thickness so that the capture of antigens by the antibodies immobilized on the brush layer could take place close to the gold SPR surface to provide a stronger optical response while the fouling was still suppressed. A minimum thickness of 19 nm of poly(HEMA) brush layer was necessary to suppress completely non-specific sensor response to fouling from milk.
- MeSH
- Enterobacteriaceae isolation & purification pathogenicity MeSH
- Antibodies, Immobilized MeSH
- Infant MeSH
- Humans MeSH
- Limit of Detection MeSH
- Milk microbiology MeSH
- Infant Formula MeSH
- Polyhydroxyethyl Methacrylate MeSH
- Food Microbiology methods statistics & numerical data MeSH
- Surface Plasmon Resonance methods statistics & numerical data MeSH
- Antibodies, Bacterial MeSH
- Gold MeSH
- Animals MeSH
- Check Tag
- Infant MeSH
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
Background and Aims: Pisum sativum L. (pea) seed is a source of carbohydrate and protein for the developing plant. By studying pea seeds inoculated by the cytokinin-producing bacterium, Rhodococcus fascians , we sought to determine the impact of both an epiphytic (avirulent) strain and a pathogenic strain on source-sink activity within the cotyledons during and following germination. Methods: Bacterial spread was monitored microscopically, and real-time reverse transcription-quantitative PCR was used to determine the expression of cytokinin biosynthesis, degradation and response regulator gene family members, along with expression of family members of SWEET , SUT , CWINV and AAP genes - gene families identified initially in pea by transcriptomic analysis. The endogenous cytokinin content was also determined. Key Results: The cotyledons infected by the virulent strain remained intact and turned green, while multiple shoots were formed and root growth was reduced. The epiphytic strain had no such marked impact. Isopentenyl adenine was elevated in the cotyledons infected by the virulent strain. Strong expression of RfIPT , RfLOG and RfCKX was detected in the cotyledons infected by the virulent strain throughout the experiment, with elevated expression also observed for PsSWEET , PsSUT and PsINV gene family members. The epiphytic strain had some impact on the expression of these genes, especially at the later stages of reserve mobilization from the cotyledons. Conclusions: The pathogenic strain retained the cotyledons as a sink tissue for the pathogen rather than the cotyledon converting completely to a source tissue for the germinating plant. We suggest that the interaction of cytokinins, CWINVs and SWEETs may lead to the loss of apical dominance and the appearance of multiple shoots.
- MeSH
- Cytokinins metabolism MeSH
- Pisum sativum genetics microbiology MeSH
- Host-Pathogen Interactions MeSH
- Germination MeSH
- Cotyledon growth & development microbiology MeSH
- Plant Diseases microbiology MeSH
- Polymerase Chain Reaction MeSH
- Rhodococcus physiology MeSH
- Seeds microbiology MeSH
- Publication type
- Journal Article MeSH
Hematooncologic patients often host rare or fastidious pathogens. Using 16S rDNA sequencing and transmission electron microscopy, we have identified 2 lymphoma patients infected with Candidatus Neoehrlichia mikurensis. In both individuals, the clinical presentation suggested ehrlichiosis-like syndrome. We believe that molecular techniques open new vistas in the field of pathogen detection.
- MeSH
- Anaplasmataceae classification genetics isolation & purification MeSH
- Molecular Diagnostic Techniques MeSH
- DNA, Bacterial genetics MeSH
- Ehrlichiosis diagnosis MeSH
- Microscopy, Electron MeSH
- Hematologic Neoplasms complications MeSH
- Fever of Unknown Origin diagnosis microbiology MeSH
- Immunocompromised Host MeSH
- Anaplasmataceae Infections complications diagnosis MeSH
- Ticks microbiology MeSH
- Middle Aged MeSH
- Humans MeSH
- Polymerase Chain Reaction MeSH
- DNA, Ribosomal genetics MeSH
- Sequence Analysis, DNA MeSH
- Animals MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Case Reports MeSH
Stále častější používání katétrů a umělých implantátů, podávání antibiotik a vysoký počet imunokompromitovaných pacientů patří mezi hlavní příčiny rostoucího významu biofilmových infekcí. Jejich charakteristické rysy, především značná rezistence k antimikrobiálním látkám a tvorba dlouhodobě perzistujících ložisek, komplikují terapii. Proto je pro klinické lékaře znalost přítomnosti tohoto faktoru patogenity cenná a měla by mít vliv na postup při léčbě pacienta. K průkazu schopnosti tvořit biofilm u klinicky významných mikroorganismů je však potřeba dostatečně spolehlivý a citlivý postup, kterého lze použít i v podmínkách běžné mikrobiologické laboratoře. Pro průkaz tohoto faktoru virulence se v současnosti používá široké spektrum vyšetřovacích technik. Vizualizací biofilmu mikroskopickými technikami počínaje přes kultivační průkaz, detekci jeho složek, odhalování fyzikálně-chemických odlišností biofilmpozitivních mikroorganismů ve srovnání s jejich planktonickými formami až po průkaz genů zodpovědných za jeho tvorbu. Vzhledem k omezením jednotlivých metod lze nejlepších výsledků dosáhnout nejlépe jejich vzájemnou kombinací.
The increasing use of catheters, artificial implants and antimicrobials as well as high numbers of immunocompromised patients are major causes for concern over biofilm infections. These infections are characterized particularly by high resistance to antimicrobials and formation of persistent foci that may complicate therapy. Therefore, detection of biofilm formation is of high relevance to the clinician and his/her approach to the treatment. Reliable and sensitive methods for detection of this pathogenicity factor in clinically important organisms, suitable for use in routine microbiological laboratories, are needed for this purpose. Currently, a wide array of techniques are available for detection of this virulence factor, such as biofilm visualization by microscopy, culture detection, detection of particular components, detection of physical and chemical differences between biofilm-positive organisms and their planktonic forms and detection of genes responsible for biofilm formation. Since each of these methods has limitations, the best results can be achieved by combining different approaches.
Many populations of the narrow-clawed crayfish Astacus leptodactylus in Turkey, including those inhabiting Lake Eğirdir, declined drastically in the mid-1980s due to introduction of crayfish plague Aphanomyces astaci. However, unlike many other localities, there has been some recovery in the A. leptodactylus population inhabiting this lake even though crayfish plague has been suspected to have persisted since then. In support of this, DNA from 5 of 34 healthy-looking crayfish sampled recently from the lake tested positive by both conventional and real-time PCR using species-specific primers targeting the rDNA internal transcribed spacer region, and product sequence analysis confirmed the identification of A. astaci. This complies with other recent reports of coexistence of native European crayfish with this pathogen, and further research is now needed to identify the key mechanisms allowing it.
- MeSH
- Population Density MeSH
- Lakes MeSH
- Polymerase Chain Reaction methods MeSH
- Astacoidea microbiology MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Turkey MeSH
