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1.
Saudi J Biol Sci ; 28(7): 4029-4038, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34220261

RESUMO

Using proteomics-based identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), we conducted the first analysis of the composition of endophytic bacteria isolated from different parts of selected Epipactis species, i.e. the buds, the inflorescences and the central part of the shoots, as well as the rhizomes. We identified aerobic and anaerobic bacteria, including such taxa as Bacillus spp., Clostridium spp., Pseudomonas spp. and Stenotrophomonas spp., which may be considered as promoting plant growth. Because most of the indicated bacteria genera belong to spore-producing taxa (spores allow bacterial symbionts to survive adverse conditions), we suggest that these bacteria species contribute to the adaptation of orchids to the environment. We found clear differences in the microbiome between investigated closely related taxa, i.e., Epipactis albensis, E. helleborine, E. purpurata and E. purpurata f. chlorophylla. Some of the analysed orchid species, i.e. E. albensis and E. purpurata co-occur in habitats, and their bacterial microbiomes differ from each other.

2.
Sci Rep ; 11(1): 6639, 2021 03 23.
Artigo em Inglês | MEDLINE | ID: mdl-33758319

RESUMO

Fungi belonging to the Cryptococcus neoformans/C. gattii species complex (CNGSC) are pathogens causing severe infections in humans and animals, that for humans may result in a mortality rate ranging up to 70%. The CNGSC is divided into eight major molecular types, that may differ in their virulence and susceptibility. In order to fully understand the epidemiology of cryptococcosis, it is important to study the world distribution and population structure of these pathogens. The present study is the first presenting a population of strains isolated in Poland and one of the few using a multi-species animal group as a source of the specimen. The pathogen was present in 2.375% of the tested animals. The URA5-RFLP and MALDI-TOF MS analyses have revealed that the population consisted exclusively of C. neoformans strains, with a predominance of major molecular type VNIV (C. neoformans var. neoformans). The MALDI-TOF MS was used to perform the CNGSC strains identification on both the species and sub-species level. Despite the fact that the animals providing the specimens were not treated with 5-fluorocytosine, around 10% of the tested population presented MIC values exceeding 64 mg/L, indicating the existence of the 5-fluorocytosine-resistant strains in the environment.


Assuntos
Doenças dos Animais/epidemiologia , Doenças dos Animais/microbiologia , Criptococose/veterinária , Cryptococcus neoformans/classificação , Doenças dos Animais/história , Animais , Antifúngicos/farmacologia , Cryptococcus neoformans/efeitos dos fármacos , Cryptococcus neoformans/genética , Cryptococcus neoformans/isolamento & purificação , História do Século XXI , Testes de Sensibilidade Microbiana , Tipagem Molecular , Técnicas de Tipagem Micológica , Polônia/epidemiologia , Polimorfismo de Fragmento de Restrição , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
3.
Int J Mol Sci ; 22(3)2021 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-33530493

RESUMO

The impact of the Gram-negative bacterium Escherichia coli (E. coli) on the microbiomic and pathogenic phenomena occurring in humans and other warm-blooded animals is relatively well-recognized. At the same time, there are scant data concerning the role of E. coli strains in the health and disease of cold-blooded animals. It is presently known that reptiles are common asymptomatic carriers of another human pathogen, Salmonella, which, when transferred to humans, may cause a disease referred to as reptile-associated salmonellosis (RAS). We therefore hypothesized that reptiles may also be carriers of specific E. coli strains (reptilian Escherichia coli, RepEC) which may differ in their genetic composition from the human uropathogenic strain (UPEC) and avian pathogenic E. coli (APEC). Therefore, we isolated RepECs (n = 24) from reptile feces and compared isolated strains' pathogenic potentials and phylogenic relations with the aforementioned UPEC (n = 24) and APEC (n = 24) strains. To this end, we conducted an array of molecular analyses, including determination of the phylogenetic groups of E. coli, virulence genotyping, Pulsed-Field Gel Electrophoresis-Restriction Analysis (RA-PFGE) and genetic population structure analysis using Multi-Locus Sequence Typing (MLST). The majority of the tested RepEC strains belonged to nonpathogenic phylogroups, with an important exception of one strain, which belonged to the pathogenic group B2, typical of extraintestinal pathogenic E. coli. This strain was part of the globally disseminated ST131 lineage. Unlike RepEC strains and in line with previous studies, a high percentage of UPEC strains belonged to the phylogroup B2, and the percentage distribution of phylogroups among the tested APEC strains was relatively homogenous, with most coming from the following nonpathogenic groups: C, A and B1. The RA-PFGE displayed a high genetic diversity among all the tested E. coli groups. In the case of RepEC strains, the frequency of occurrence of virulence genes (VGs) was lower than in the UPEC and APEC strains. The presented study is one of the first attempting to compare the phylogenetic structures of E. coli populations isolated from three groups of vertebrates: reptiles, birds and mammals (humans).


Assuntos
Doenças dos Animais/microbiologia , Infecções por Escherichia coli/veterinária , Filogenia , Répteis/microbiologia , Escherichia coli Uropatogênica/classificação , Escherichia coli Uropatogênica/genética , Animais , Proteínas de Escherichia coli/genética , Especificidade de Hospedeiro , Humanos , Tipagem de Sequências Multilocus , Doenças das Aves Domésticas/microbiologia , Virulência/genética , Fatores de Virulência/genética
4.
Acta Biochim Pol ; 67(3): 393-399, 2020 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-32945647

RESUMO

We compared fecal samples from responders and non-responders to administration of Lactobacillus reuteri DSM 17938. Data for this post hoc analysis were collected from an RCT assessing the efficacy of L. reuteri for the management of acute gastroenteritis. Responders were defined as subjects with diarrhea lasting no longer than 48 h. 44 children (17 responders and 27 non-responders) were analyzed. There were no differences in clinical characteristics and gut colonization between both groups. In the responder group, there were significantly lower levels of five metabolites before beginning of the intervention: lactate, choline, ethanol, creatine, and formate. The fecal calprotectin level did not differ between groups prior to the intervention, but its level was significantly lower after intervention in the responder group. Possibly, the responder group with a "metabolic niche", including lower level of metabolites, especially lactate, that are potential products of Lactobacillus genus, would determine the response to probiotic treatment. These findings need to be confirmed, but identification of some differences in the fecal metabolomics and the calprotectin level suggests that further studies are warranted.


Assuntos
Diarreia/dietoterapia , Suplementos Nutricionais/microbiologia , Fezes/química , Fezes/microbiologia , Gastroenterite/dietoterapia , Lactobacillus reuteri/metabolismo , Probióticos/uso terapêutico , Doença Aguda , Pré-Escolar , Diarreia/metabolismo , Método Duplo-Cego , Feminino , Gastroenterite/metabolismo , Humanos , Lactente , Complexo Antígeno L1 Leucocitário/análise , Complexo Antígeno L1 Leucocitário/metabolismo , Masculino , Metaboloma , Resultado do Tratamento
5.
Curr Microbiol ; 77(9): 2166-2171, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32424607

RESUMO

Reptiles appear to be an important vector for Gram-negative pathogens, therefore, they are epidemiologically relevant. However, the composition of reptilian microbiota has been poorly recognized so far. The majority of studies concern exotic reptiles as asymptomatic carriers of Salmonella serovars. Studies of other intestinal bacteria of reptiles are rare. Only recently, the microbiota of free-living European reptiles have been investigated, however, on the basis of small samples, mainly in protected areas. Here, we aim to investigate cloacal Gram-negative microbiota of free-living Natrix natrix. Snakes (N = 45) used in the study were collected in Kraków (Poland) and its vicinity. Nineteen species of Gram-negative bacteria were isolated. The most common species were: Aeromonas hydrophila, Morganella morganii, Proteus vulgaris, Salmonella spp. The bacteria prevalent in N. natrix cloacal swabs are likely to represent the natural intestinal Gram-negative microbiota of the examined snakes. Importantly, the identified bacteria are pathogenic to humans, which clearly highlights the epidemiological potential of free-living N. natrix. The risk of infection is high for immunocompromised humans, children (under 5 years old), elderly persons, and pregnant women. Our study provides the largest dataset on intestinal Gram-negative microbiota of wild snakes. The presence of multiple human pathogens determined by us calls for the necessity of further studies on reptile-transmitted bacteria in anthropogenic environments.


Assuntos
Colubridae , Microbiota , Idoso , Animais , Criança , Pré-Escolar , Feminino , Humanos , Polônia , Gravidez , Salmonella
6.
Int J Mol Sci ; 21(8)2020 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-32294944

RESUMO

The genus Lactobacillus includes, among others, Lactobacillus casei, Lactobacillus paracasei and Lactobacillus rhamnosus, species that are collectively referred to as the Lactobacillus casei group. Many studies have shown that strains belonging to this group may decrease lactose intolerance, the effects of inflammatory bowel disease, diarrhea, constipation, food allergies and even colon cancer. Moreover, evidences exists of positive effects of these bacteria on mucosal immunity and blood cholesterol level. Because of their beneficial influence on human health, many of them are used as food additives and probiotic pharmaceuticals. It should be stressed that health-promoting properties are not attributed at the species level, but to specific strains. Therefore, procedures are necessary to allow specific identification at each phylogenetic level-genus, species and strain. In this paper we present a practical overview of molecular methods for the identification and differentiation of L. casei bacteria. The research included 30 bacterial strains belonging to three species: L.casei, L. paracasei and L. rhamnosus. Among the tested procedures were genus- and species-specific PCR, multiplex-PCR, Real-Time HRM analysis, RFLP-PCR, rep-PCR, RAPD-PCR, AFLP-PCR, and proteomic methods such as MALDI-TOF MS typing and SDS-PAGE fingerprinting. The obtained results showed that multiplex-PCR and MALDI-TOF MS turned out to be the most useful methods to identify the tested bacteria at the species level. At the strain level, the AFLP-PCR method showed the highest discriminatory power. We hope that the presented results will allow for the easy selection of an appropriate procedure, depending on the experiment conducted and the equipment capabilities of any given laboratory.


Assuntos
Lactobacillus casei/classificação , Lactobacillus casei/genética , Tipagem Molecular , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , DNA Bacteriano , Humanos , Tipagem Molecular/métodos , Reação em Cadeia da Polimerase Multiplex , Reação em Cadeia da Polimerase , Probióticos
7.
Future Microbiol ; 15: 259-271, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32271108

RESUMO

Aim: Characterization of the ability of Fusobacterium nucleatum DSM 15643 and DSM 20482 strains in the presence of Cu2+ and H2O2 to reactive oxygen species generation. Method: Spectrophotometric ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) method was used. Results: Determination of: MIC for Cu2+, H2O2 and ABTS; survivability of F. nucleatum under atmospheric oxygen exposure; the level and rate constants of free radicals production by the bacteria. Conclusion: F. nucleatum in the presence of Cu2+ and H2O2 is able to generate free radicals. Reactive oxygen species are produced mainly outside the bacterial cell, which suggests that outer membrane proteins may be involved in oxidative process.


Assuntos
Fusobacterium nucleatum/química , Fusobacterium nucleatum/metabolismo , Espécies Reativas de Oxigênio/análise , Espécies Reativas de Oxigênio/metabolismo , Espectrofotometria/métodos , Benzotiazóis/química , Cobre/farmacologia , Fusobacterium nucleatum/efeitos dos fármacos , Peróxido de Hidrogênio/farmacologia , Ácidos Sulfônicos/química
8.
BMC Vet Res ; 15(1): 312, 2019 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-31477105

RESUMO

BACKGROUND: Salmonella is generally considered as a human pathogen causing typhoid fever and gastrointestinal infections called salmonellosis, with S. Enteritidis and S. Typhimurium strains as the main causative agents. Salmonella enterica strains have a wide host array including humans, birds, pigs, horses, dogs, cats, reptiles, amphibians and insects. Up to 90% of reptiles are the carriers of one or more serovars of Salmonella. Extraintestinal bacterial infections associated with reptiles pose serious health threat to humans. The import of exotic species of reptiles as pet animals to Europe correlates with the emergence of Salmonella serotypes, which not found previously in European countries. The presented study is a new report about Salmonella serotypes associated with exotic reptiles in Poland. The goal of this research was to examine the zoonotic potential of Salmonella strains isolated from reptiles by comparative analysis with S. Enteritidis strains occurring in human population and causing salmonellosis. RESULTS: The main findings of our work show that exotic reptiles are asymptomatic carriers of Salmonella serovars other than correlated with salmonellosis in humans (S. Enteritidis, S. Typhimurium). Among the isolated Salmonella strains we identified serovars that have not been reported earlier in Poland, for example belonging to subspecies diarizonae and salamae. Restriction analysis with Pulsed-field Gel Electrophoresis (PFGE), showed a great diversity among Salmonella strains isolated from reptiles. Almost all tested strains had distinct restriction patterns. While S. Enteritidis strains were quite homogeneous in term of phylogenetic relations. Most of the tested VGs were common for the two tested groups of Salmonella strains. CONCLUSIONS: The obtained results show that Salmonella strains isolated from reptiles share most of virulence genes with the S. Enteritidis strains and exhibit a greater phylogenetic diversity than the tested S. Enteritidis population.


Assuntos
Eletroforese em Gel de Campo Pulsado , Répteis/microbiologia , Infecções por Salmonella/microbiologia , Salmonella enterica/genética , Animais , Portador Sadio , Cromatografia Gasosa , DNA Bacteriano , Genótipo , Humanos , Salmonella enterica/patogenicidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrometria de Massas em Tandem , Virulência , Zoonoses
9.
J Vet Diagn Invest ; 31(4): 523-530, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31006359

RESUMO

We compared the effectiveness of various methods for the identification of Staphylococcus spp. other than S. aureus isolated from intramammary infections of cows on 3 dairy farms in Lower Silesia, Poland. A total of 131 isolates belonging to 18 Staphylococcus species were identified by sequence analysis of the 16S rRNA and dnaJ genes, as well using a commercial identification system (ID 32 STAPH; bioMérieux) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS; Bruker Daltonics). Sequencing of the 16S rRNA gene was found to have low discriminatory value because only 43% of isolates were recognized unequivocally. Much better results were obtained with the dnaJ gene (all isolates were correctly identified at the species level). However, some of these isolates achieved a low similarity level (<97%) and required a confirmatory test (sequencing of the rpoB gene). The performance of ID 32 STAPH was poor. Regardless of the probability level used (80% or 90%), the commercial system obtained identification rates <40%. Using MALDI-TOF MS and the commercial Bruker database, 67% of isolates were identified correctly with scores ≥2.0 (acceptable species-level identification) but this number increased to 97% after the database was expanded. The definitive identification of Staphylococcus spp. other than S. aureus causing intramammary infections in cattle often requires a combination of different procedures, and the existing databases should be updated.


Assuntos
Mastite Bovina/microbiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterinária , Infecções Estafilocócicas/veterinária , Staphylococcus/classificação , Staphylococcus/genética , Animais , Bovinos , Feminino , Genótipo , Humanos , Mastite Bovina/epidemiologia , Polônia/epidemiologia , RNA Ribossômico 16S/genética , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia
11.
BMC Microbiol ; 18(1): 86, 2018 08 17.
Artigo em Inglês | MEDLINE | ID: mdl-30119648

RESUMO

BACKGROUND: Yersinia enterocolitica is widespread within the humans, pigs and wild boars. The low isolation rate of Y. enterocolitica from food or environmental and clinical samples may be caused by limited sensitivity of culture methods. The main goal of present study was identification of presumptive Y. enterocolitica isolates using MALDI TOF MS. The identification of isolates may be difficult due to variability of bacterial strains in terms of biochemical characteristics. This work emphasizes the necessity of use of multiple methods for zoonotic Y. enterocolitica identification. RESULTS: Identification of Y. enterocolitica isolates was based on MALDI TOF MS, and verified by VITEK® 2 Compact and PCR. There were no discrepancies in identification of all human' and pig' isolates using MALDI TOF MS and VITEK® 2 Compact. However three isolates from wild boars were not decisively confirmed as Y. enterocolitica. MALDI TOF MS has identified the wild boar' isolates designated as 3dz, 4dz, 8dz as Y. enterocolitica with a high score of matching with the reference spectra of MALDI Biotyper. In turn, VITEK® 2 Compact identified 3dz and 8dz as Y. kristensenii, and isolate 4dz as Y. enterocolitica. The PCR for Y. enterocolitica 16S rDNA for these three isolates was negative, but the 16S rDNA sequence analysis identified these isolates as Y. kristensenii (3dz, 4dz) and Y. pekkanenii (8dz). The wild boar' isolates 3dz, 4dz and 8dz could not be classified using biotyping. The main bioserotype present within pigs and human faeces was 4/O:3. It has been shown that Y. enterocolitica 1B/O:8 can be isolated from human faeces using ITC/CIN culturing. CONCLUSION: The results of our study indicate wild boars as a reservoir of new and atypical strains of Yersinia, for which protein and biochemical profiles are not included in the MALDI Biotyper or VITEK® 2 Compact databases. Pigs in the south-west Poland are the reservoir for pathogenic Y. enterocolitica strains. Four biochemical features included in VITEK® 2 Compact known to be common with Wauters scheme were shown to produce incompatible results, thus VITEK® 2 Compact cannot be applied in biotyping of Y. enterocolitica.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Sus scrofa/microbiologia , Suínos/microbiologia , Yersinia enterocolitica/isolamento & purificação , Animais , DNA Ribossômico , Reservatórios de Doenças/microbiologia , Fezes/microbiologia , Humanos , Polônia , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 16S/genética , Análise de Sequência , Especificidade da Espécie , Yersinia/classificação , Yersinia/genética , Yersinia/isolamento & purificação , Yersinia enterocolitica/classificação , Yersinia enterocolitica/genética
12.
Vet Microbiol ; 214: 28-35, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29408029

RESUMO

The aim of this paper is to describe a novel subpopulation of Staphylococcus haemolyticus isolated from intramammary gland infections (IMI) in cattle. In total, eight isolates originating from milk samples from two unrelated dairy farms were examined phenotypically (using the ID 32 STAPH system) and genotypically. These isolates had almost identical sequences of each of the housekeeping genes examined (dnaJ, rpoB and sodA) but these sequences displayed similarity of only ∼92.5%, 95.0% and 96.8%, respectively, with known S. haemolyticus sequences. The atypical isolates could also be distinguished biochemically by the positive ß-galactosidase test (with 2-naphthyl-ß-d-galactopyranoside as the substrate). All the isolates were identified as S. haemolyticus upon MALDI-TOF analysis but half of them, that achieved scores 1.7-1.999 (not reliable species identification), required expanding the commercial database for secure identification. Our study has shown that IMI in cattle may be caused by two distinct subpopulations of S. haemolyticus, differing clearly by some genotypic and phenotypic properties. The first of these subpopulations seems to be common to many hosts (including humans), whereas the second (possibly at the subspecies rank) is, so far, found only in cattle.


Assuntos
Inflamação/microbiologia , Leite/microbiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus haemolyticus/genética , Staphylococcus haemolyticus/isolamento & purificação , Animais , Bovinos , Indústria de Laticínios , Feminino , Genótipo , Glândulas Mamárias Animais/microbiologia , Mastite Bovina/microbiologia , Fenótipo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Staphylococcus haemolyticus/classificação
13.
Microb Drug Resist ; 24(3): 314-322, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28628752

RESUMO

Campylobacter spp. is a major cause of foodborne diseases in humans, particularly when transmitted by the handling or consumption of undercooked poultry meat. Most Campylobacter infections are self-limiting, but antimicrobial treatment (e.g., fluoroquinolones and macrolides) is necessary in severe or prolonged cases. The indiscriminate use of these drugs, both in clinical medicine and animal production, has a major impact on public health. The aim of the present study was to identify Campylobacter strains, isolated from turkey and broilers, using both PCR and the matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) methods to reveal the accuracy of identification, as well to evaluate the antimicrobial and genetic resistance of the investigated strains. MALDI-TOF and PCR methods were used to show differences, if any, in the specificity of that test. In this study, MALDI-TOF mass spectrometry gave the same results as multiplex PCR, in all cases. The highest rate of resistance (i.e., 100% of turkey and broiler strains) was detected against ciprofloxacin, whereas 58.1% of turkey and 78.6% of broiler strains were resistant to tetracycline. Multidrug-resistant isolates were not found in the study. All ciprofloxacin-resistant strains had a mutation in the gyrA gene, at the Thr-86 position. The presence of the tetO gene was found in 71% of turkey and in 100% of broiler strains. All resistant to tetracycline strains included tetO gene. Additionally, in five turkey and three broiler strains, susceptible to tetracycline, tetO gene was present. These results indicate the high prevalence of Campylobacter strains, which are phenotypically and genetically resistant to fluoroquinolones and tetracycline.


Assuntos
Antibacterianos/farmacologia , Infecções por Campylobacter/veterinária , Campylobacter coli/genética , Campylobacter jejuni/genética , Farmacorresistência Bacteriana/genética , Doenças das Aves Domésticas/epidemiologia , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Infecções por Campylobacter/tratamento farmacológico , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/microbiologia , Campylobacter coli/classificação , Campylobacter coli/efeitos dos fármacos , Campylobacter coli/isolamento & purificação , Campylobacter jejuni/classificação , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/isolamento & purificação , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Galinhas/microbiologia , Ciprofloxacina/farmacologia , DNA Girase/genética , DNA Girase/metabolismo , Fazendas , Fezes/microbiologia , Expressão Gênica , Humanos , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase Multiplex/veterinária , Polônia/epidemiologia , Aves Domésticas/microbiologia , Doenças das Aves Domésticas/tratamento farmacológico , Doenças das Aves Domésticas/microbiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterinária , Tetraciclina/farmacologia , Perus/microbiologia
14.
BMC Infect Dis ; 17(1): 763, 2017 12 12.
Artigo em Inglês | MEDLINE | ID: mdl-29233117

RESUMO

BACKGROUND: Elizabethkingia miricola is a rare Gram-negative bacterium found in water and clinical specimens. Typical culturing methods often misidentify Elizabethkingia spp. as Flavobacterium or Chryseobacterium. Although diagnosis is based on culturing samples taken from sterile sites, such as blood, a proper identification of this bacterium requires an expertise that goes beyond the capabilities of a typical clinical laboratory. CASE PRESENTATION: A 35-year-old woman diagnosed with common variable immunodeficiency was admitted to our center. Previous treatment with antibiotics (amoxicillin plus clavulanate, first and third generation of cephalosporins, macrolides) and systemic corticosteroids (up to 120 mg/day of prednisolone) failed to arrest the spread of inflammation. Gingival recession was observed in her oral cavity, resulting in an apparent lengthening of her teeth. In addition to typical commensal bacteria, including streptococci and neisseriae, strains of Rothia mucilaginosa and Elizabethkingia miricola were identified upon a detailed microbiological examination using a MALDI-TOF MS Biotyper system. The presence of the latter strain correlated with severe periodontitis, lack of IgA in her saliva and serum, a very low IgG concentration (< 50 mg/dl), IgM-paraproteinemia, decreases in C3a and C5a and microvascular abnormality. High-dose immunoglobulin (to maintain IgG > 500 mg/dl) and targeted levofloxacin treatment resulted in immune system reconstitution, oral healing, and eradication of the Elizabethkingia infection. CONCLUSIONS: E. miricola rarely causes disease in healthy individuals. However, the overgrowth of commensal bacteria, lack of IgG/IgA, microvasculopathy and complement cascade activation in patients with humoral immunodeficiency may facilitate Elizabethkingia invasion. Overuse of antibiotics, particularly beta-lactams, may cause mucosal colonization by E. miricola, followed by its multiplication combined with periodontitis that prompts bacterial translocation. MALDI-TOF Biotyper analysis may become a method of choice for identification of Elizabethkingia infections.


Assuntos
Infecções por Bactérias Gram-Negativas/diagnóstico , Periodontite/diagnóstico , Corticosteroides/uso terapêutico , Adulto , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Complemento C3a/análise , Complemento C5a/análise , Feminino , Flavobacteriaceae/efeitos dos fármacos , Flavobacteriaceae/genética , Flavobacteriaceae/isolamento & purificação , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/imunologia , Humanos , Imunidade Humoral , Imunoglobulina A/análise , Imunoglobulina A/sangue , Imunoglobulinas Intravenosas/uso terapêutico , Levofloxacino/uso terapêutico , Boca/microbiologia , Periodontite/tratamento farmacológico , Periodontite/imunologia , RNA Ribossômico 16S/isolamento & purificação , RNA Ribossômico 16S/metabolismo , Saliva/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
15.
Artigo em Inglês | MEDLINE | ID: mdl-28924447

RESUMO

BACKGROUND: Allergic, especially anaphylactic, reactions during immunoglobulin replacement therapy are rare, but their pathophysiology and classification remain ambiguous. Recent findings show positive results of skin tests with commercially available immunoglobulins, but target antigens and responsible compounds of the tested immunoglobulins have not been strictly identified. CASE DESCRIPTION AND FINDINGS: Four adult patients with recently diagnosed common variable immunodeficiency qualified for standard subcutaneous immunoglobulin replacement therapy regimen. They had no history of receiving immunoglobulins, blood or blood product transfusions. Edema, confluent wheals and erythema were observed at the site of subcutaneous immunoglobulin infusion: typical early and late phase reaction. A transient increase in various passively transferred IgG and IgE antibodies was responsible for misleading positive outcome of the serological testing for active humoral response such as type I allergy, anti-Rh, isohemagglutinins and rheumatoid factor (RF). Although the clinical presentation was very unusual and severe, the retrospective analysis showed no isohemagglutinins, RF and IgE in the patients' serum before but it was positive after the infusion (median IgE = 18 IU/ml, RF = 8 IU/ml). Type I allergic reaction (laryngeal edema, rhinoconjuctivitis) came out at +14 days of replacement therapy when the patient visited countryside. In the second patient anaphylactic reaction was observed 5 days after ScIg administration, and only when the patient consumed peanuts. Therefore, IgE concentration was measured retrospectively in a series of commercial preparations used in the initial subcutaneous immunoglobulin replacement therapy that caused the adverse event (AE) and it was determined between 138 and 232 IU/ml (kU/l), i.e. 690-2100 IU per g of protein. Specific IgE was within a wide range from 198 (mix of food) to 2809 kUA/l (mix of grass) but many of the tested allergen-specific IgE were class 2 or 3 (i.e. 0.71-17.5 kUA/l). CONCLUSIONS: The case resembles passive cutaneous anaphylaxis and Prausnitz-Küstner reaction but clinical significance of the classical phenomena has not yet been described. This observation indicates that anaphylactic reactions during immunoglobulin replacement therapy may result from IgE or pathological IgG content. Such IgE presence was sporadically reported (34.5-105 IU/ml, i.e. 862.5-1450 IU/g of protein) in intravenous immunoglobulins that are used and monitored by healthcare professionals. In clinical practice the definition of adverse events is inadequate since individual batches of immunoglobulins come with different specificity therefore, they should be classified as transfusion products (not bioequivalents). Such new approach implies establishing (1) new control methods and strategies to ensure introduction of the safety regulations for subcutaneous home self-administration of immunoglobulins as well as (2) guidelines for the prevention of anaphylaxis in patients receiving immunoglobulins (for example peanut).

17.
BMC Microbiol ; 17(1): 169, 2017 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-28750622

RESUMO

BACKGROUND: Citrobacter strains are opportunistic pathogens often responsible for serious enteric as well as extra-intestinal diseases, and therefore the O-antigenic scheme, still in use in diagnostic identification, should be set for proper serotyping. The structures of more than 30 different Citrobacter O-antigens (O-polysaccharide chains of the lipopolysaccharides) of 43 Citrobacter O-serogroups have been elucidated so far. However, relationships between strains in several heterogeneous serogroups still need to be clarified by immunochemical studies. These include complex serogroups O3 and O8, represented by 20 and 7 strains, respectively, which are the subject of the present work. Earlier, the O-polysaccharide structures have been determined for Citrobacter O3 strain Be35/57 (PCM 1508) and Citrobacter O8 strain Be64/57 (PCM 1536). RESULTS: Serological studies (immunoblotting) carried out on Citrobacter lipopolysaccharides from different strains ascribed to serogroups O3 and O8 showed that each of these serogroups should be divided into non-cross-reacting subgroups. Based on the results of chemical analyses and 1H and 13C NMR spectroscopy the structure of Citrobacter O-antigens from strains PCM 1504 (O6) and PCM 1573 (O2) have been established. Chemical data combined with serological analyses showed that several Citrobacter strains should be reclassified into other serogroups. CONCLUSIONS: Immunochemical studies carried out on Citrobacter LPS, described in this paper, showed the expediency of reclassification of: 1) strains PCM 1504 and PCM 1573 from serogroups O6 and O2 to serogroups O3 and O8, respectively, 2) strains PCM 1503 and PCM 1505 from serogroups O3 and O8 to new serogroups O3a and O8a, respectively.


Assuntos
Citrobacter/classificação , Infecções por Enterobacteriaceae/microbiologia , Citrobacter/genética , Citrobacter/imunologia , Citrobacter/isolamento & purificação , Humanos , Lipopolissacarídeos/química , Lipopolissacarídeos/imunologia , Espectroscopia de Ressonância Magnética , Filogenia , Sorogrupo
19.
Nat Microbiol ; 1: 16027, 2016 03 21.
Artigo em Inglês | MEDLINE | ID: mdl-27572446

RESUMO

Together with plague, smallpox and typhus, epidemics of dysentery have been a major scourge of human populations for centuries(1). A previous genomic study concluded that Shigella dysenteriae type 1 (Sd1), the epidemic dysentery bacillus, emerged and spread worldwide after the First World War, with no clear pattern of transmission(2). This is not consistent with the massive cyclic dysentery epidemics reported in Europe during the eighteenth and nineteenth centuries(1,3,4) and the first isolation of Sd1 in Japan in 1897(5). Here, we report a whole-genome analysis of 331 Sd1 isolates from around the world, collected between 1915 and 2011, providing us with unprecedented insight into the historical spread of this pathogen. We show here that Sd1 has existed since at least the eighteenth century and that it swept the globe at the end of the nineteenth century, diversifying into distinct lineages associated with the First World War, Second World War and various conflicts or natural disasters across Africa, Asia and Central America. We also provide a unique historical perspective on the evolution of antibiotic resistance over a 100-year period, beginning decades before the antibiotic era, and identify a prevalent multiple antibiotic-resistant lineage in South Asia that was transmitted in several waves to Africa, where it caused severe outbreaks of disease.


Assuntos
Disenteria Bacilar/epidemiologia , Disenteria Bacilar/microbiologia , Evolução Molecular , Filogeografia , Sorogrupo , Shigella dysenteriae/classificação , Shigella dysenteriae/isolamento & purificação , Farmacorresistência Bacteriana , Disenteria Bacilar/história , Genoma Bacteriano , Saúde Global , História do Século XIX , História do Século XX , História do Século XXI , Humanos , Epidemiologia Molecular , Análise de Sequência de DNA , Shigella dysenteriae/genética
20.
J Basic Microbiol ; 56(8): 922-8, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26972384

RESUMO

The pathogenicity of entomopathogenic nematodes (EPNs) depends directly on the presence of bacteria in the nematode digestive tracts. Based on 16S rRNA and MALDI-TOF analyses 20 isolated bacteria were assigned to 10 species with 10 isolates classified as Pseudomonas ssp. Six strains (30%) show ureolytic activity on Christensen medium. Spectroscopic analysis of the strains showed that the ureolytic activity is strongly correlated with the following wavenumbers: 935 cm(-1) in window W4, which carries information about the bacterial cell wall construction and 1158 cm(-1) in window W3 which corresponds to proteins in bacterial cell. A logistic regression model designed on the basis of the selected wavenumbers differentiates ureolytic from non-ureolytic bacterial strains with an accuracy of 100%. Spectroscopic studies and mathematical analyses made it possible to differentiate EPN-associated Pseudomonas sp. strains from clinical Pseudomonas aeruginosa PAO1. These results suggest, that infrared spectra of EPN-associated Pseudomonas sp. strains may reflect its adaptation to the host.


Assuntos
Insetos/parasitologia , Nematoides/microbiologia , Pseudomonas/metabolismo , Ureia/metabolismo , Animais , Pseudomonas/genética , Pseudomonas/isolamento & purificação
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