Clostridium perfringens forms biofilms and spores that are a source of food contamination. In this study, the antibacterial activities of Lactobacillus plantarum culture supernatants (LP-S), LP-S fractions, and the plant-derived compound epigallocatechin gallate (EG) were evaluated. Specifically, their effects on the viability and biofilm-forming ability of C. perfringens were assessed. Moreover, the expression of quorum sensing-regulated genes associated with the pathogenesis of this microorganism and that of genes involved in biofilm formation was also investigated. The results showed that both EG and the LP-S exerted bactericidal activity against all C. perfringens strains tested. The minimal bactericidal concentration (MBC) of EG was 75 µg/mL for all strains but ranged from 61 to 121 µg of total protein per mL for LP-S. EG exerted only minor effects on biofilm formation, whereas LP-S, particularly its 10 and 30 K fractions, significantly reduced the biofilm-forming ability of all the strains. The antibiofilm activity of LP-S was lost following preincubation with proteases, suggesting that it was mediated by a proteinaceous molecule. The treatment of C. perfringens with either EG or LP-S did not change the transcript levels of two CpAL (C. perfringens quorum-sensing Agr-like system)-related genes, agrB and agrD, which are known to be involved in the regulation of biofilms, suggesting that LP-S exerted its biofilm inhibitory activity downstream of CpAL signaling. In summary, we demonstrated the bactericidal activity of EG and LP-S against C. perfringens and antibiofilm activity of LP-S at a subinhibitory dose. Our results suggested that these compounds can be further explored for food safety applications to control agents such as C. perfringens.
- MeSH
- biofilmy MeSH
- Clostridium perfringens * účinky léků genetika MeSH
- katechin analogy a deriváty farmakologie MeSH
- kultivační média speciální * farmakologie MeSH
- Lactobacillus plantarum * metabolismus MeSH
- regulace genové exprese u bakterií účinky léků MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- epigallocatechin gallate MeSH Prohlížeč
- katechin MeSH
- kultivační média speciální * MeSH
Targeted proteomics recently proved to be a technique for the detection and absolute quantification of proteins not easily accessible to classical bottom-up approaches. Due to this, it has been considered as a high fidelity tool to detect potential warfare agents in wide spread kinds of biological and environmental matrices. Clostridium perfringens toxins are considered to be potential biological weapons, especially the epsilon toxin which belongs to a group of the most powerful bacterial toxins. Here, the development of a target mass spectrometry method for the detection of C. perfringens protein toxins (alpha, beta, beta2, epsilon, iota) is described. A high-resolution mass spectrometer with a quadrupole-Orbitrap system operating in target acquisition mode (parallel reaction monitoring) was utilized. Because of the lack of commercial protein toxin standards recombinant toxins were prepared within Escherichia coli. The analysis was performed using proteotypic peptides as the target compounds together with their isotopically labeled synthetic analogues as internal standards. Calibration curves were calculated for each peptide in concentrations ranging from 0.635 to 1101 fmol/μL. Limits of detection and quantification were determined for each peptide in blank matrices.
- Klíčová slova
- Clostridium perfringens, PRM, epsilon toxin, mass spectrometry, protein toxin,
- MeSH
- bakteriální proteiny analýza genetika MeSH
- bakteriální toxiny analýza genetika MeSH
- chromatografie kapalinová MeSH
- Clostridium perfringens * genetika růst a vývoj metabolismus MeSH
- Escherichia coli genetika MeSH
- peptidy analýza genetika MeSH
- proteomika MeSH
- rekombinantní proteiny analýza MeSH
- tandemová hmotnostní spektrometrie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- bakteriální proteiny MeSH
- bakteriální toxiny MeSH
- peptidy MeSH
- rekombinantní proteiny MeSH
Anaerobic bacteria, such as Bacteroides fragilis or Clostridium perfringens, are part of indigenous human flora. However, Clostridium difficile represents also an important causative agent of nosocomial infectious antibiotic-associated diarrhoea. Treatment of C. difficile infection is problematic, making it imperative to search for new compounds with antimicrobial properties. Hops (Humulus lupulus L.) contain substances with antibacterial properties. We tested antimicrobial activity of purified hop constituents humulone, lupulone and xanthohumol against anaerobic bacteria. The antimicrobial activity was established against B. fragilis, C. perfringens and C. difficile strains according to standard testing protocols (CLSI, EUCAST), and the minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBC) were calculated. All C. difficile strains were toxigenic and clinically relevant, as they were isolated from patients with diarrhoea. Strongest antimicrobial effects were observed with xanthohumol showing MIC and MBC values of 15-107 μg/mL, which are close to those of conventional antibiotics in the strains of bacteria with increased resistance. Slightly higher MIC and MBC values were obtained with lupulone followed by higher values of humulone. Our study, thus, shows a potential of purified hop compounds, especially xanthohumol, as alternatives for treatment of infections caused by select anaerobic bacteria, namely nosocomial diarrhoea caused by resistant strains.
- Klíčová slova
- Clostridium, Xanthohumol, anaerobic bacteria, gut,
- MeSH
- anaerobióza fyziologie MeSH
- antibakteriální látky izolace a purifikace farmakologie MeSH
- Bacteroides fragilis účinky léků růst a vývoj MeSH
- Clostridioides difficile účinky léků růst a vývoj patogenita MeSH
- Clostridium perfringens účinky léků růst a vývoj MeSH
- cyklohexeny izolace a purifikace farmakologie MeSH
- flavonoidy izolace a purifikace farmakologie MeSH
- Humulus chemie MeSH
- infekce spojené se zdravotní péčí mikrobiologie MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- propiofenony izolace a purifikace farmakologie MeSH
- průjem mikrobiologie MeSH
- pseudomembranózní enterokolitida mikrobiologie MeSH
- rostlinné extrakty chemie MeSH
- symbióza fyziologie MeSH
- terpeny izolace a purifikace farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antibakteriální látky MeSH
- cyklohexeny MeSH
- flavonoidy MeSH
- humulon MeSH Prohlížeč
- lupulon MeSH Prohlížeč
- propiofenony MeSH
- rostlinné extrakty MeSH
- terpeny MeSH
- xanthohumol MeSH Prohlížeč
Clostridium perfringens is a Validated Biological Agent and a pathogen of medical, veterinary, and military significance. Gas gangrene is the most destructive of all the clostridial diseases and is caused by C. perfringens type A strains wherein the infection spreads quickly (several inches per hour) with production of gas. Influence of repeated in vitro cultivation on the infectivity of C. perfringens was investigated by comparing the surface proteins of laboratory strain and repository strains of the bacterium using 2DE-MS approach. In order to optimize host-pathogen interaction during experimental gas gangrene infection, we also explored the role of particulate matrix on ability of C. perfringens to cause gas gangrene.
- MeSH
- bakteriální proteiny genetika metabolismus MeSH
- Clostridium perfringens genetika růst a vývoj fyziologie MeSH
- gangréna plynatá mikrobiologie MeSH
- interakce hostitele a patogenu MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- bakteriální proteiny MeSH
Clostridium perfringens-induced necrotic enteritis is generally controlled by antibiotics. However, because of increasing antibiotic resistance, other antibacterial agents are required, preferably ones that do not affect the beneficial intestinal microbiota of the host. This study evaluated the in vitro selective growth-inhibitory effect of 8-hydroxyquinoline (8HQ) on C. perfringens vs. bifidobacteria in a medium containing chicken ileal digesta. Prior to the experiments, the minimum inhibitory concentrations of 8HQ and penicillin G were determined by broth microdilution assay. The minimum inhibitory concentration values of 8HQ for C. perfringens were 16-32 times lower than the values for bifidobacteria. Treatment of autoclaved and non-autoclaved chicken ileal digesta with 8HQ showed a selective anticlostridial effect. After incubation of C. perfringens with autoclaved ileal digesta for 3 h, all 8HQ concentrations tested (32-2048 μg/mL) significantly reduced C. perfringens bacterial count. In contrast, the same treatment had no or only a slight effect on bifidobacteria counts. Unlike 8HQ, penicillin G did not exhibit any selectivity. Similar results were obtained after incubation for 24 h. In non-autoclaved ileal digesta, all 8HQ concentrations tested significantly reduced C. perfringens bacterial counts after incubation for 30 min and 3 h, while no effect was observed on bifidobacteria. These results suggest that 8HQ may serve as a prospective veterinary compound for use against necrotic enteritis in poultry.
- MeSH
- antibakteriální látky farmakologie MeSH
- Bifidobacterium účinky léků MeSH
- Clostridium perfringens účinky léků MeSH
- ileum mikrobiologie MeSH
- klostridiové infekce prevence a kontrola veterinární virologie MeSH
- kur domácí mikrobiologie MeSH
- nemoci drůbeže prevence a kontrola virologie MeSH
- oxychinolin farmakologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antibakteriální látky MeSH
- oxychinolin MeSH
The aim of this study was to assess the effect of a complete nitrite replacement strategy using celery, carmine, sodium lactate and orange dietary fibre combined with vitamins C and E, on the quality characteristics (technological, sensorial and safety properties) of hot dog sausages (five samples) during chilled storage (2 ± 1℃ 60 days). Nitrite replacers (combined with vitamins C and E) presented antioxidant activity, reducing lipid oxidation in reformulated samples. At the end of storage redness (a*) was similar in the control sample (with added nitrite) and in the sample without added nitrite. Sensory evaluation detected no significant difference between samples with and without added nitrite. All the reformulated samples were judged acceptable by the panellists. At the end of storage, the control sample contained more than four times as much residual nitrite as the reformulated samples. Growth of presumptive Clostridium perfringens was not observed in any of the samples. Samples without added nitrite had longer shelf-lives than control sausage. Samples containing 0.1% vitamin C registered the lowest microbiological levels. This strategy could be a good alternative to reduce and/or eliminate added nitrite in hot dog sausages.
- Klíčová slova
- Hot dog, L-ascorbic acid, orange dietary fibre, residual nitrite, sodium lactate, vitamin E,
- MeSH
- antiinfekční látky farmakologie MeSH
- antioxidancia farmakologie MeSH
- Apium MeSH
- chuť * MeSH
- Citrus chemie MeSH
- Clostridium perfringens účinky léků růst a vývoj MeSH
- dusitany * MeSH
- karmín farmakologie MeSH
- konzervace potravin metody MeSH
- kyselina askorbová farmakologie MeSH
- lidé MeSH
- manipulace s potravinami metody MeSH
- masné výrobky analýza mikrobiologie MeSH
- natriumlaktát farmakologie MeSH
- peroxidace lipidů účinky léků MeSH
- potravinářská mikrobiologie MeSH
- potravní vláknina MeSH
- skladování potravin metody MeSH
- skot MeSH
- vitamin E farmakologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antiinfekční látky MeSH
- antioxidancia MeSH
- dusitany * MeSH
- karmín MeSH
- kyselina askorbová MeSH
- natriumlaktát MeSH
- potravní vláknina MeSH
- vitamin E MeSH
Infection of prosthetic hemodialysis (HD) shunts is one of the most common complications of vascular access in hemodialysis patients. The incidence of anaerobic infection is very rare. In such a case, management of treatment represents a great challenge for the surgeon. We report a complicated case of autologous hemodialysis shunt infected by Clostridium perfringens on the right forearm in a polymorbid female patient with chronic renal failure and myelodysplastic syndrome. The patient has undergone repeated establishment of HD shunt with subsequent polybacterial local infectious complications. Destructive clostridial infection developed at the site of the infectious complications. This very rare complication involving infection by an atypical bacterial agent requires rapid diagnosis and comprehensive multimodal treatment including surgical, antibiotic and supportive therapy. Clinical implications and optimal therapy is not precisely defined in these cases.
A prerequisite for successful identification of anaerobic pathogenic bacteria from samples of clinical material is the method of cultivation. Currently, several methods of cultivation in anaerobic environment are used: cultivation in anaerobic box, anaerobic jar, and in nonrecurring cultivation system. Here, we determined the suitability of the above methods of cultivation using the estimation of the growth (diameters of colony size) of commonly isolated anaerobic pathogens (Bacteroides fragilis, Clostridium difficile, and Clostridium perfringens). The tested bacterial strains were exposed to atmospheric oxygen for various time periods and then they were cultivated using different anaerobic cultivation systems. Maximum growth differed, depending on the type of cultivation and the strain used. Thus, largest zone diameters, in the majority of measurements, were achieved in the anaerobic box. However, nonrecurring cultivation system seemed better in several cases; this applied to the cultivation of C. perfringens after 15, 30, and 60 min exposure to atmospheric oxygen as well as the cultivation of B. fragilis after 30 and 60 min of oxygen exposure. The cultivation in anaerobic box was the most convenient method for growth of C. difficile. In almost all cases, higher growth was observed in nonrecurring cultivation system than in the system of anaerobic jar. On the other hand, no significant differences were observed among these anaerobic cultivation systems which confirmed their applicability (taking into account some individual features concerning the optimization of cultivations) for identification of pathogenic anaerobes.
- MeSH
- anaerobióza MeSH
- anaerobní bakterie metabolismus MeSH
- Bacteroides fragilis metabolismus MeSH
- Clostridioides difficile metabolismus MeSH
- Clostridium perfringens metabolismus MeSH
- klostridiové infekce mikrobiologie MeSH
- kyslík metabolismus MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- kyslík MeSH
Bilirubin is degraded in the human gut by microflora into urobilinoids. In our study we investigated whether the bilirubin-reducing strain of Clostridium perfringens can reduce bilirubin ditaurate (BDT), a bile pigment of some lower vertebrates, without hydrolysis of the taurine moiety. C. perfringes was incubated under anaerobic conditions with BDT; reduction products were quantified by spectrophotometry and separated by TLC. Based on Rf values of BDT reduction products and synthetic urobilinogen ditaurate, three novel taurine-conjugated urobilinoids were identified. It is likely that bilirubin-reducing enzyme(s) serve for the effective disposal of electrons produced by fermentolytic processes in these anaerobic bacteria.
- MeSH
- bilirubin analogy a deriváty izolace a purifikace metabolismus MeSH
- chromatografie na tenké vrstvě MeSH
- Clostridium perfringens izolace a purifikace metabolismus MeSH
- feces mikrobiologie MeSH
- hydrolýza MeSH
- lidé MeSH
- novorozenec MeSH
- oxidace-redukce MeSH
- střeva mikrobiologie MeSH
- taurin analogy a deriváty izolace a purifikace metabolismus MeSH
- urobilinogen MeSH
- Check Tag
- lidé MeSH
- novorozenec MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- bilirubin ditaurine MeSH Prohlížeč
- bilirubin MeSH
- taurin MeSH
- urobilinogen MeSH
