Immunohistological Localization of the MrkD Adhesin in the Type 3 Fimbriae of Klebsiella pneumoniae

1998 ◽  
Vol 66 (5) ◽  
pp. 2356-2361 ◽  
Author(s):  
Ann-Mari Tarkkanen ◽  
Benita Westerlund-Wikström ◽  
Leena Erkkilä ◽  
Timo K. Korhonen
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Fusion Protein ◽  
Polyclonal Antibodies ◽  
Western Blots ◽  
Deletion Derivative ◽  
E Coli ◽  
Minor Protein ◽  
Type 3

ABSTRACT The adhesive minor protein MrkD of the type 3 fimbria ofKlebsiella pneumoniae was expressed and purified fromEscherichia coli as a fusion protein with an N-terminal polyhistidine tail. Polyclonal antibodies raised against MrkD specifically recognized the MrkD peptide in Western blots of fimbrial preparations. Immunoelectron microscopic analyses showed that the anti-MrkD immunoglobulins bound to the tip of the plasmid-encoded variant of the type 3 fimbria of K. pneumoniae, whereas no binding to the chromosomally encoded MrkD-deficient type 3 fimbrial variant of K. pneumoniae was detected. Immunoglobulins from an antiserum raised against purified type 3 fimbrial filaments bound laterally to both type 3 fimbrial variants. The anti-MrkD antibodies also bound to the tip of a papG deletion derivative of theE. coli P fimbria complemented with mrkD, indicating that MrkD structurally complements a PapG mutation in the P fimbria of E. coli.

scholarly journals Identification of Type 3 Fimbriae in Uropathogenic Escherichia coli Reveals a Role in Biofilm Formation

2007 ◽  
Vol 190 (3) ◽  
pp. 1054-1063 ◽  
Author(s):  
Cheryl-Lynn Y. Ong ◽  
Glen C. Ulett ◽  
Amanda N. Mabbett ◽  
Scott A. Beatson ◽  
Richard I. Webb ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Klebsiella Oxytoca ◽  
The United States ◽  
Conjugative Plasmid ◽  
Biofilm Growth ◽  
Indwelling Catheters ◽  
E Coli ◽  
Type 3

ABSTRACT Catheter-associated urinary tract infection (CAUTI) is the most common nosocomial infection in the United States. Uropathogenic Escherichia coli (UPEC), the most common cause of CAUTI, can form biofilms on indwelling catheters. Here, we identify and characterize novel factors that affect biofilm formation by UPEC strains that cause CAUTI. Sixty-five CAUTI UPEC isolates were characterized for phenotypic markers of urovirulence, including agglutination and biofilm formation. One isolate, E. coli MS2027, was uniquely proficient at biofilm growth despite the absence of adhesins known to promote this phenotype. Mini-Tn5 mutagenesis of E. coli MS2027 identified several mutants with altered biofilm growth. Mutants containing insertions in genes involved in O antigen synthesis (rmlC and manB) and capsule synthesis (kpsM) possessed enhanced biofilm phenotypes. Three independent mutants deficient in biofilm growth contained an insertion in a gene locus homologous to the type 3 chaperone-usher class fimbrial genes of Klebsiella pneumoniae. These type 3 fimbrial genes (mrkABCDF), which were located on a conjugative plasmid, were cloned from E. coli MS2027 and could complement the biofilm-deficient transconjugants when reintroduced on a plasmid. Primers targeting the mrkB chaperone-encoding gene revealed its presence in CAUTI strains of Citrobacter koseri, Citrobacter freundii, Klebsiella pneumoniae, and Klebsiella oxytoca. All of these mrkB-positive strains caused type 3 fimbria-specific agglutination of tannic acid-treated red blood cells. This is the first description of type 3 fimbriae in E. coli, C. koseri, and C. freundii. Our data suggest that type 3 fimbriae may contribute to biofilm formation by different gram-negative nosocomial pathogens.

scholarly journals Immunological Characterization of Escherichia coli O157:H7 Intimin γ1

2002 ◽  
Vol 9 (1) ◽  
pp. 46-53 ◽  
Author(s):  
W.-G. Son ◽  
T. A. Graham ◽  
V. P. J. Gannon
Keyword(s):  
Escherichia Coli ◽  
Fusion Protein ◽  
Full Length ◽  
Expression Vectors ◽  
Protein Fusion ◽  
Western Blots ◽  
E Coli ◽  
Specific Pcr ◽  
His Tag

ABSTRACT Portions of the intimin genes of Escherichia coli O157:H7 strain E319 and of the enteropathogenic E. coli O127:H6 strain E2348/69 were amplified by PCR and cloned into pET-28a(+) expression vectors. The entire 934 amino acids (aa) of E. coli O157:H7 intimin, the C-terminal 306 aa of E. coli O157:H7 intimin, and the C-terminal 311 aa of E. coli O127:H6 intimin were expressed as proteins fused with a six-histidine residue tag (six-His tag) in pET-28a(+). Rabbit antisera raised against the six-His tag-full-length E. coli O157:H7 intimin protein fusion cross-reacted in slot and Western blots with outer membrane protein preparations from the majority of enterohemorrhagic and enteropathogenic E. coli serotypes which have the intimin gene. The E. coli strains tested included isolates from humans and animals which produce intimin typesα (O serogroups 86, 127, and 142), β1 (O serogroups 5, 26, 46, 69, 111, 126, and 128), γ1 (O serogroups 55, 145, and 157), γ2 (O serogroups 111 and 103), and ε (O serogroup 103) and a nontypeable intimin (O serogroup 80), results based on intimin type-specific PCR assays. Rabbit antisera raised against the E. coli O157:H7 C-terminal fusion protein were much more intimin type-specific than those raised against the full-length intimin fusion protein, but some cross-reaction with other intimin types was also observed for these antisera. In contrast, the monoclonal antibody Intγ1.C11, raised against the C-terminal E. coli O157 intimin, reacted only with preparations from intimin γ1-producing E. coli strains such as E. coli O157:H7.

Антимікробна резистентність провідних збудників інфекцій сечових шляхів у Києві (Україна)

2017 ◽  
Vol 1 (2) ◽  
pp. 48-60
Author(s):  
A.G. Salmanov ◽  
A.V. Rudenko
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Klebsiella Pneumoniae ◽  
Staphylococcus Epidermidis ◽  
Klebsiella Oxytoca ◽  
Enterobacter Aerogenes ◽  
Enterobacter Cloacae ◽  
Proteus Vulgaris ◽  
Providencia Rettgeri ◽  
E Coli

Мета роботи — вивчити резистентність до антибіотиків бактеріальних збудників інфекцій сечових шляхів (ІСШ), виділених у пацієнтів урологічного стаціонару в м. Києві. Матеріали і методи. Досліджено 1612 штамів бактерій, виділених із сечі хворих з ІСШ (цистит, уретрит, пієлонефрит), госпіталізованих в урологічне відділення ДУ «Інститут урології НАМН України» у м. Києві протягом 2016 р. Серед пацієнтів переважали жінки — 1201 (74,5 %). Вік хворих становив від 17 до 74 років. Для збору даних використано медичну документацію лікарні. Мікробіологічні дослідження виконано у лабораторії мікробіології ДУ «Інститут урології НАМН України». Аналізували результати культурального дослідження зразків сечі, зібраних за наявності клінічних ознак ІСШ. Дослідження клінічного матеріалу та інтерпретацію отриманих результатів проводили загальноприйнятими методами. Вивчено чутливість уропатогенів до 31 антибіотика дискодифузійним методом відповідно до рекомендацій Інституту клінічних та лабораторних стандартів США (Clinical and Laboratory Standards Institute (CLSI)). Результати та обговорення. Аналіз мікробного спектра сечі виявив домінування серед уропатогенів штамів Escherichia coli (32,0 %), Enterococcus faecalis (19,5 %), Klebsiella pneumoniae (10,9 %), Staphylococcus epidermidis (8,9 %), S. haemolyticus (6,5 %) та Pseudomonas aeruginosa (6,4 %). Частка Enterococcus faecium, Enterobacter aerogenes і Streptococcus viridans становила відповідно 2,5, 2,2 і 1,6 %, Enterobacter cloacae, Klebsiella oxytoca, Acinetobacter baumannii, Proteus vulgaris та Providencia rettgeri — менше 1,0 %. У більшості випадків (69,7 %) мікроорганізми виділено у монокультурі, у решті випадків — у мікробних асоціа- ціях. Високу резистентність до тестованих антибіотиків виявили штами E. aerogenes (45,1 %), E. cloacae (45,7 %), E. faecium (40,9 %), E. faecalis (40,7 %), E. coli (39,9 %), P. aeruginosa (34,0 %), K. pneumoniae (28,6 %). Найбільш активними до уропатогенів були іміпенем (E. coli — 87,6 %, P. aeruginosa — 75,7 %, E. cloacae — 67,3 %, E. aerogenes — 72,6 %, K. pneumoniae — 93,2 %), меропенем (E. coli — 89,1 %, P. aeruginosa — 76,7 %, K. pneumoniae — 82,6 %), лефлоцин (E. coli — 74,5 %, ентерококи — 78,7 %, P. aeruginosa — 76,7 %, E. cloacae — 73,9 %, E. aerogenes — 80,4 %, K. pneumoniae — 83,5 %), амоксицилін/клавуланат (ентерококи — 84,6 %), фурагін (ентерококи — 82,6 %), цефоперазон (K. pneumoniae — 89,2 %, P. aeruginosa — 73,8 %), цефтріаксон (K. pneumoniae — 80,1 %). Висновки. Антибіотикорезистентність збудників ІСШ — важлива терапевтична проблема. Найбільшою активністю до уропатогенів характеризуються іміпенем, меропенем, лефлоцин, амоксицилін/ клавуланат, фурагін, цефоперазон, цефтріаксон, які можна розглядати як препарат вибору для призначення стартової терапії ІСШ. Необхідно здійснювати постійний моніторинг за резистентністю до дії антибіотиків. Політику використання антибіотиків у кожному стаціонарі слід визначати залежно від локальних даних щодо резистентності до протимікробних препаратів.

scholarly journals Prevalence and Characterization of Extended-Spectrum β-Lactamase-Producing Antibiotic-Resistant Escherichia coli and Klebsiella pneumoniae in Ready-to-Eat Street Foods

Antibiotics ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 850
Author(s):  
Shobha Giri ◽  
Vaishnavi Kudva ◽  
Kalidas Shetty ◽  
Veena Shetty
Keyword(s):  
Escherichia Coli ◽  
Food Safety ◽  
Klebsiella Pneumoniae ◽  
Rural Areas ◽  
Significant Risk ◽  
Microbiological Quality ◽  
Resistant Bacteria ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Extended Spectrum

As the global urban populations increase with rapid migration from rural areas, ready-to-eat (RTE) street foods are posing food safety challenges where street foods are prepared with less structured food safety guidelines in small and roadside outlets. The increased presence of extended-spectrum-β-lactamase (ESBL) producing bacteria in street foods is a significant risk for human health because of its epidemiological significance. Escherichia coli and Klebsiella pneumoniae have become important and dangerous foodborne pathogens globally for their relevance to antibiotic resistance. The present study was undertaken to evaluate the potential burden of antibiotic-resistant E. coli and K. pneumoniae contaminating RTE street foods and to assess the microbiological quality of foods in a typical emerging and growing urban suburb of India where RTE street foods are rapidly establishing with public health implications. A total of 100 RTE food samples were collected of which, 22.88% were E. coli and 27.12% K. pneumoniae. The prevalence of ESBL-producing E. coli and K. pneumoniae was 25.42%, isolated mostly from chutneys, salads, paani puri, and chicken. Antimicrobial resistance was observed towards cefepime (72.9%), imipenem (55.9%), cefotaxime (52.5%), and meropenem (16.9%) with 86.44% of the isolates with MAR index above 0.22. Among β-lactamase encoding genes, blaTEM (40.68%) was the most prevalent followed by blaCTX (32.20%) and blaSHV (10.17%). blaNDM gene was detected in 20.34% of the isolates. This study indicated that contaminated RTE street foods present health risks to consumers and there is a high potential of transferring multi-drug-resistant bacteria from foods to humans and from person to person as pathogens or as commensal residents of the human gut leading to challenges for subsequent therapeutic treatments.

Expression and partial purification of human pro-opiomelanocortin in Escherichia coli

1989 ◽  
Vol 3 (2) ◽  
pp. 105-112 ◽  
Author(s):  
T. S. Grewal ◽  
P. J. Lowry ◽  
D. Savva
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Fusion Protein ◽  
Western Blot ◽  
Blot Analysis ◽  
Expression Vectors ◽  
Partial Purification ◽  
Amino Acid Residues ◽  
E Coli ◽  
Dna Fragment

ABSTRACT A large portion of the human pro-opiomelanocortin (POMC) peptide corresponding to amino acid residues 59–241 has been cloned and expressed in Escherichia coli. A 1·0 kb DNA fragment encoding this peptide was cloned into the expression vectors pUC8 and pUR291. Plasmid pJMBG51 (a pUC8 recombinant) was found to direct the expression of a 24 kDa peptide. The recombinant pUR291 (pJMBG52) was shown to produce a β-galactosidase fusion protein of 140 kDa. Western blot analysis showed that both the 24 kDa and 140 kDa peptides are recognized by antibodies raised against POMC-derived peptides. The β-galactosidase fusion protein has been partially purified from crude E. coli cell lysates using affinity chromatography on p-aminobenzyl-1-thio-β-d-galactopyranoside agarose.

Resistencia antibiótica de Escherichia coli y Staphylococcus saprophyticus en la infección urinaria de un hospital público

2021 ◽  
Vol 61 (4) ◽  
pp. 633-641
Author(s):  
Jarvis Raraz-Vidal ◽  
Henry Allpas-Gomez ◽  
Omar Raraz-Vidal
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
E Coli ◽  
Staphylococcus Saprophyticus ◽  
De Medicina

Introducción: La resistencia de antibióticos puede llegar a causar una amplia morbilidad y complicaciones. Objetivo: Determinar el perfil de resistencia antimicrobiana de Escherichia Coli y de Staphylococcus Saprophyticus, en pacientes con infección urinaria hospitalizados en el servicio de Medicina Interna del Hospital Municipal Los Olivos. Métodos: Estudio descriptivo, retrospectivo de corte transversal. Se realizó en el servicio de Medicina Interna del Hospital Municipal los Olivos (HMLO). Participantes: historia clínica de pacientes hospitalizados con infección urinaria en el servicio de Medicina Interna. Intervenciones: Según los criterios de inclusión y exclusión se obtuvieron, 96 historias clínicas (HC) del año 2013. Se utilizó un instrumento de recolección validado. Se realizó el análisis descriptivo con software estadístico STATA versión 25. Resultados: De las 96 HC, la edad promedio fue 55,04 años, los agentes microbianos más frecuentes fueron: la Escherichia coli con 85,3%, Staphylococcus saprophyticus 4.2% y Klebsiella pneumoniae 3,1%. La prevalencia de productores de betalactamasa espectro extendido (BLEE) fue 10,4%. Los antibióticos más resistentes fueron: trimetoprim/sulfametoxazol 89,6%, ampicilina 86%, piperacilina 84,6%, tetraciclina 79,2% y ciprofloxacino 70,8%. Los antibióticos más sensibles fueron: amikacina 100%, imipenem 100%, ertapenem 98%, meropenem 96% y piperacilina/tazobactam 96%. Conclusión: El uropatógeno más frecuente en pacientes con ITU hospitalizados fue la E. coli. Los antibióticos que presentaron resistencia a la E. coli fueron: trimetoprim/sulfametoxazol, ampicilina, piperacilina, tetraciclina y ciprofloxacino, y para el S. Saprophyticus fueron: amoxicilina/ ácido clavulánico, trimetoprim/sulfametoxazol, ceftriaxona y ciprofloxacino.

scholarly journals Incidence and Antimicrobial Susceptibility of Escherichia coli and Klebsiella pneumoniae with Extended-Spectrum β-Lactamases in Community- and Hospital-Associated Intra-Abdominal Infections in Europe: Results of the 2008 Study for Monitoring Antimicrobial Resistance Trends (SMART)

2010 ◽  
Vol 54 (7) ◽  
pp. 3043-3046 ◽  
Author(s):  
Stephen P. Hawser ◽  
Samuel K. Bouchillon ◽  
Daryl J. Hoban ◽  
Robert E. Badal ◽  
Rafael Cantón ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Klebsiella Pneumoniae ◽  
Antimicrobial Susceptibility ◽  
Beta Lactamase ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Extended Spectrum ◽  
Abdominal Infections

ABSTRACT From 2002 to 2008, there was a significant increase in extended-spectrum beta-lactamase (ESBL)-positive Escherichia coli isolates in European intra-abdominal infections, from 4.3% in 2002 to 11.8% in 2008 (P < 0.001), but not for ESBL-positive Klebsiella pneumoniae isolates (16.4% to 17.9% [P > 0.05]). Hospital-associated isolates were more common than community-associated isolates, at 14.0% versus 6.5%, respectively, for E. coli (P < 0.001) and 20.9% versus 5.3%, respectively, for K. pneumoniae (P < 0.01). Carbapenems were consistently the most active drugs tested.

Cloning and expression of Xylella fastidiosa antigens in Escherichia coli and Erwinia stewartii

1989 ◽  
Vol 35 (4) ◽  
pp. 487-491 ◽  
Author(s):  
Paul H. Goodwin
Keyword(s):  
Escherichia Coli ◽  
Molecular Weight ◽  
Genomic Dna ◽  
Xylella Fastidiosa ◽  
Gene Bank ◽  
Cloning And Expression ◽  
Western Blots ◽  
E Coli ◽  
Cosmid Vector ◽  
Erwinia Stewartii

Xylella fastidiosa DNA, partially digested with Sau3A, was ligated into the cosmid vector, pUCD615. Approximately 4500 ampicillin-resistant Escherichia coli colonies were obtained. The frequency of complementation of leucine auxotrophy in transfected E. coli indicated that the cosmid gene bank was representative of X. fastidiosa genomic DNA. Colonies were lysed directly onto nitrocellulose membranes using a thermo-inducible λ lysogen and screened for expression of X. fastidiosa antigens. Approximately 16.5% of a random sample of clones were found to express X. fastidiosa antigens as determined by Western blots. These proteins comigrated with proteins of X. fastidiosa and ranged in molecular weight from 10 000 to 160 000. Conjugation of several of the plasmids into Erwinia stewartii resulted in expression of the similar molecular weight cloned proteins with similar levels of expression as in E. coli.Key words: Xylella fastidiosa, Pierce's disease, immunological clone screening, thermo-inducible lysogeny.

scholarly journals Antimicrobial Resistance Patterns and ESBL of Uropathogens Isolated from Adult Females in Najran Region of Saudi Arabia

2021 ◽  
Vol 11 (3) ◽  
pp. 650-658
Author(s):  
Mohammed Yahia Alasmary
Keyword(s):  
Escherichia Coli ◽  
Saudi Arabia ◽  
Antimicrobial Resistance ◽  
Klebsiella Pneumoniae ◽  
Urine Samples ◽  
Klebsiella Species ◽  
E Coli ◽  
Government Hospitals ◽  
Resistance Patterns ◽  
Tract Infections

Background: To explore the prevalence of urinary tract infections (UTIs) among female patients in the Najran region of Saudi Arabia and determine their antimicrobial resistance pattern. Methods: This study was conducted on 136 urine samples collected from outpatient departments (OPDs) of the different government hospitals in the Najran region of Saudi Arabia. Over one year, the results of susceptibility testing reports of outpatient midstream urine samples from three government hospitals were prospectively evaluated. Results: Of 136 urine samples, only 123 (90.45%) were found to show significant growth for UTIs, from which 23 different uropathogens were identified. Escherichia coli (58.5%) was the most commonly isolated organism, followed by Klebsiella pneumoniae (8.1%). The isolated microorganism showed increased resistance patterns from 3.3% to 62.6%, with an overall resistance of 27.19%. Meropenem was the most effective antimicrobial, followed by amikacin and ertapenem (0.47%, 0.91%, and 1.5% resistance, respectively). At the same time, ampicillin and cephazolin were the least (62.6% and 59.5% resistance, respectively) effective. Overall, eleven (8.94%) uropathogens isolates were ESBLs, among which there were eight (6.5%) Escherichia coli, one (0.81%) Klebsiella pneumoniae, one (0.81%) Klebsiella oxytoca, and one (0.81%) Citrobacter amalonaticus. Conclusions: E. coli remains the most commonly isolated causative uropathogens, followed by Klebsiella species. The prevalence of pathogenic E. coli and Klebsiella species underscores the importance of developing cost-effective, precise, and rapid identification systems to minimize public exposure to uropathogens. Antibiotic susceptibility data revealed that most of the isolates were resistant to the majority of the antibiotics. The patients with UTIs in the Najran region of Saudi Arabia are at a high risk of antibiotic resistance, leading to significant problems in outpatient department (OPD) treatment outcomes and raising the alarm for the physician to change their empiric treatment.

scholarly journals Bactérias gram-negativas em cardeais (Paroaria coronata e Paroaria dominicana) apreendidos do tráfico de animais silvestres

2016 ◽  
Vol 53 (1) ◽  
pp. 107 ◽  
Author(s):  
Marcos Paulo Vieira Cunha ◽  
Marta Brito Guimarães ◽  
Yamê Miniero Davies ◽  
Liliane Milanelo ◽  
Terezinha Knöbl
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Klebsiella Pneumoniae ◽  
Sao Paulo ◽  
São Paulo ◽  
Salmonella Spp ◽  
E Coli

Anualmente o tráfico de animais silvestres retira milhões de aves da natureza. Os cardeais (Paroaria coronata) e cardeais-do-nordeste (Paroaria dominicana) estão incluídos entre as espécies de aves mais traficadas. A microbiota cloacal de passeriformes de vida livre é composta principalmente por bacilos e cocos gram-positivos, já os bacilos gram-negativos predominam em aves de cativeiro. Em situações de estresse e baixa de imunidade as bactérias gram-negativas podem causar infecções oportunistas. O presente trabalho identificou bactérias da microbiota da cloaca de 49 espécimes de P. coronata e P. dominicana apreendidas do tráfico de animais silvestres em São Paulo (SP). Foram isoladas treze espécies de bactérias gram-negativas, incluindo Salmonella spp. e Pseudomonas aeruginosa. A maior frequência de ocorrência foi de Escherichia coli, identificada em 42/49 (85,7%) das amostras fecais. Dentre os isolados de E. coli, 21/42 pertenciam aos grupos filogenéticos B2 e D, relacionados a estirpes patogênicas que causam doença extraintestinal em humanos. Klebsiella pneumoniae foi isolada em 28/49 (57,1%) das amostras. Esses resultados reforçam que as condições estressantes a que esses animais são submetidos em situações de tráfico, incluindo o contato com humanos, podem favorecer a colonização da microbiota cloacal das aves por patógenos, o que representa um risco para a sua reintrodução na natureza considerando-se o possível contato com humanos e outros animais.

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