Science.gov

Sample records for klebsiella pneumoniae genotpiseerimine

  1. Endogenous Klebsiella pneumoniae endophthalmitis.

    PubMed

    Yin, Wenpeng; Zhou, Haijiang; Li, Chunsheng

    2014-10-01

    Klebsiella pneumonia is a common human pathogen, and endogenous endophthalmitis is a vision-threatening infection presentedwith pain, redness, decreased vision acuity, and intraocular inflammation. Endogenous endophthalmitis caused by Klebsiella pneumoniae is uncommon and usually happens in patients with immunosuppression conditions. Diabetes is a predisposing risk factor, and liver abscess is a major source of Klebsiella pneumonia endogenous endophthalmitis (KPEE). Here, we report a case of KPEE in a patient who lost his vision in one eye after treatment.

  2. Klebsiella pneumoniae Flocculation Dynamics

    PubMed Central

    Jackson, T. L.; Taylor, K. A.; Thompson, A. P.; Younger, J. G.

    2011-01-01

    The bacterial pathogen Klebsiella pneumoniae is a cause of community- and hospital-acquired lung, urinary tract, and blood stream infections. A common contaminant of indwelling catheters, it is theorized that a common infection pathway for this organism is via shedding of aggregates off of biofilm colonies. In an effort to better understand bacterial proliferation in the host bloodstream, we develop a PDE model for the flocculation dynamics of Klebsiella pneumoniae in suspension. Existence and uniqueness results are provided, as well as a brief description of the numerical approximation scheme. We generate artificial data and illustrate the requirements to accurately identify proliferation, aggregation, and fragmentation of flocs in the experimental domain of interest. PMID:18071828

  3. Chronic Klebsiella pneumonia: a rare manifestation of Klebsiella pneumonia.

    PubMed

    Boonsarngsuk, Viboon; Thungtitigul, Poungrat; Suwatanapongched, Thitiporn

    2015-09-01

    K. pneumoniae can present as two forms of community-acquired pneumonia, acute and chronic. Although acute pneumonia may turn into necrotizing pneumonia, which results in a prolonged clinical course, it often has a rapidly progressive clinical course. In contrast, chronic Klebsiella pneumonia runs a protracted indolent course that mimics other chronic pulmonary infections and malignancies. Herein, we present two cases of chronic Klebsiella pneumonia. The diagnosis was made by microorganism identification, as well as absence of other potential causes. Clinical and radiographic findings improved after a prolonged course of antibiotic therapy.

  4. Hypervirulent (hypermucoviscous) Klebsiella pneumoniae

    PubMed Central

    Shon, Alyssa S.; Bajwa, Rajinder P.S.; Russo, Thomas A.

    2013-01-01

    A new hypervirulent (hypermucoviscous) variant of Klebsiella pneumoniae has emerged. First described in the Asian Pacific Rim, it now increasingly recognized in Western countries. Defining clinical features are the ability to cause serious, life-threatening community-acquired infection in younger healthy hosts, including liver abscess, pneumonia, meningitis and endophthalmitis and the ability to metastatically spread, an unusual feature for enteric Gram-negative bacilli in the non-immunocompromised. Despite infecting a healthier population, significant morbidity and mortality occurs. Although epidemiologic features are still being defined, colonization, particularly intestinal colonization, appears to be a critical step leading to infection. However the route of entry remains unclear. The majority of cases described to date are in Asians, raising the issue of a genetic predisposition vs. geospecific strain acquisition. The traits that enhance its virulence when compared with “classical” K. pneumoniae are the ability to more efficiently acquire iron and perhaps an increase in capsule production, which confers the hypermucoviscous phenotype. An objective diagnostic test suitable for routine use in the clinical microbiology laboratory is needed. If/when these strains become increasingly resistant to antimicrobials, we will be faced with a frightening clinical scenario. PMID:23302790

  5. Endogenous Klebsiella endophthalmitis associated with Klebsiella pneumoniae pneumonia.

    PubMed

    Chen, Kuan-Jen; Hwang, Yih-Shiou; Chen, Yen-Po; Lai, Chi-Chun; Chen, Tun-Lu; Wang, Nan-Kai

    2009-01-01

    To investigate the management, bacterial strains, antibiotic sensitivities, and visual outcomes in patients with Klebsiella pneumoniae pneumonia and endogenous Klebsiella endophthalmitis. Data were collected for treatments, antibiotic sensitivity patterns, and final visual outcomes. The study included 10 eyes of 9 patients with a median age of 42 years (range, 0-86 years). Diabetes mellitus was the most common comorbid risk factor (n = 5, 56%). Nine eyes (90%) were treated with intravitreal antibiotics, and one with pars plana vitrectomy and intravitreal antibiotics. One eye achieved a favorable visual acuity of 20/20; however, 6 eyes developed vision of no light perception, including 2 of evisceration. Two nosocomial K. pneumoniae isolates were extended-spectrum-beta-lactamase-producing strains, which demonstrated the resistance to amikacin and ceftazidime. Ophthalmologists and physicians should be aware of Klebsiella pneumonia as a possible cause of endogenous endophthalmitis, and endogenous Klebsiella endophthalmitis usually causes poor visual outcomes.

  6. Klebsiella pneumoniae inoculants for enhancing plant growth

    DOEpatents

    Triplett, Eric W.; Kaeppler, Shawn M.; Chelius, Marisa K.

    2008-07-01

    A biological inoculant for enhancing the growth of plants is disclosed. The inoculant includes the bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101, Pantoea agglomerans P102, Klebsiella pneumoniae 342, Klebsiella pneumoniae zmvsy, Herbaspirillum seropedicae Z152, Gluconacetobacter diazotrophicus PA15, with or without a carrier. The inoculant also includes strains of the bacterium Pantoea agglomerans and K. pneumoniae which are able to enhance the growth of cereal grasses. Also disclosed are the novel bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101 and P102, and Klebsiella pneumoniae 342 and zmvsy.

  7. Osteopontin promotes host defense during Klebsiella pneumoniae-induced pneumonia.

    PubMed

    van der Windt, G J W; Hoogerwerf, J J; de Vos, A F; Florquin, S; van der Poll, T

    2010-12-01

    Klebsiella pneumoniae is a common cause of nosocomial pneumonia. Osteopontin (OPN) is a phosphorylated glycoprotein involved in inflammatory processes, some of which is mediated by CD44. The aim of this study was to determine the role of OPN during K. pneumoniae-induced pneumonia. Wild-type (WT) and OPN knockout (KO) mice were intranasally infected with 10⁴ colony forming units of K. pneumoniae, or administered Klebsiella lipopolysaccharides (LPS). In addition, recombinant OPN (rOPN) was intranasally administered to WT and CD44 KO mice. During Klebsiella pneumonia, WT mice displayed elevated pulmonary and plasma OPN levels. OPN KO and WT mice showed similar pulmonary bacterial loads 6 h after infection; thereafter, Klebsiella loads were higher in lungs of OPN KO mice and the mortality rate in this group was higher than in WT mice. Early neutrophil recruitment into the bronchoalveolar space was impaired in the absence of OPN after intrapulmonary delivery of either Klebsiella bacteria or Klebsiella LPS. Moreover, rOPN induced neutrophil migration into the bronchoalveolar space, independent from CD44. In vitro, OPN did not affect K. pneumoniae growth or neutrophil function. In conclusion, OPN levels were rapidly increased in the bronchoalveolar space during K. pneumoniae pneumonia, where OPN serves a chemotactic function towards neutrophils, thereby facilitating an effective innate immune response.

  8. [Construction of polyhydroxybutyrate pathway in Klebsiella pneumoniae].

    PubMed

    Guo, Xiaochen; Liu, Hongjuan; Wang, Yanping; Zhang, Jian'an; Liu, Dehua

    2013-10-01

    1,3-propanediol production with the byproduct of biodiesel production is important to increase the economic benefit of biodiesel industry. Accumulation of 3-hydroxypropionaldehyde is one of the key problems in the 1,3-propanediol fermentation process, leading to the cell death and the fermentation abnormal ceasing. Different from the traditional way of reducing the accumulation of the 3-hydroxypropionaldehyde, we introduced the polyhydroxybutyrate pathway into the Klebsiella pneumoniae for the first time to enhance the tolerance of K. pneumoniae to 3-hydroxypropionaldehyde, at the same time, to improve the 1,3-propanediol production. Plasmid pDK containing phbC, phbA, phbB gene was constructed and transformed into K. pneumoniae successfully. PHB was detected in the engineered K. pneumoniae after IPTG induction and its content enhanced with the IPTG concentration increasing. The optimized IPTG concentration was 0.5 mmol/L. The constructed K. pneumoniae could produce 1,3-propanediol normally, at the same time accumulate polyhydroxybutyrate. With the constructed strain, the fermentation proceeds normally with the initial glucose was 70 g/L which the wild type strain stopped growing and the fermentation was ceasing; 1,3-propanediol concentration and yield reached 31.3 g/L and 43.9% at 72 h. Our work is helpful for the deep understanding of 1,3-propanediol metabolic mechanism of Klebsiella pneumoniae, and also provides a new way for strain optimization of Klebsiella pneumoniae.

  9. No Carbapenem Resistance in Pneumonia Caused by Klebsiella Species

    PubMed Central

    Yayan, Josef; Ghebremedhin, Beniam; Rasche, Kurt

    2015-01-01

    Abstract Klebsiella species are a common cause of community- and nosocomial-acquired pneumonia. Antibiotic resistance to the class of carbapenem in patients with pneumonia caused by Klebsiella species is unusual. New studies report carbapenem resistance in patients with pneumonia caused by Klebsiella species. This article examines, retrospectively, antibiotic resistance in patients with community- and nosocomial-acquired pneumonia caused by Klebsiella species. The data of all patients with community- and nosocomial-acquired pneumonia caused by Klebsiella species were collected from the hospital charts at the HELIOS Clinic, Witten/Herdecke University, Wuppertal, Germany, within the study period 2004 to 2014. An antibiogram was created from all of the study patients with pneumonia caused by Klebsiella species. Sensitivity and resistance profiles were performed for the different antibiotics that have been consistently used in the treatment of patients with pneumonia caused by Klebsiella species. All demographic, clinical, and laboratory data of all of the patients with pneumonia caused by Klebsiella species were collected from the patients’ records. During the study period of January 1, 2004, to August 12, 2014, 149 patients were identified with community- and nosocomial-acquired pneumonia affected by Klebsiella species. These patients had a mean age of 70.6 ± 13 (107 [71.8%, 95% CI 64.6%–79%] men and 42 [28.2%, 95% CI 21%–35.4%] women). In all of the patients with pneumonia caused by Klebsiella species, there was resistance to ampicillin (P < 0.0001). Many patients with pneumonia caused by Klebsiella species (75.3%) also showed resistance to piperacillin (P < 0.0001). However, no patients with pneumonia caused by Klebsiella species showed resistance to imipenem or meropenem (P < 0.0001). Antibiotic resistance to the antibiotic class of carbapenem was not detected in patients with pneumonia caused by Klebsiella species. PMID:25674753

  10. No carbapenem resistance in pneumonia caused by Klebsiella species.

    PubMed

    Yayan, Josef; Ghebremedhin, Beniam; Rasche, Kurt

    2015-02-01

    Klebsiella species are a common cause of community- and nosocomial-acquired pneumonia. Antibiotic resistance to the class of carbapenem in patients with pneumonia caused by Klebsiella species is unusual. New studies report carbapenem resistance in patients with pneumonia caused by Klebsiella species.This article examines, retrospectively, antibiotic resistance in patients with community- and nosocomial-acquired pneumonia caused by Klebsiella species.The data of all patients with community- and nosocomial-acquired pneumonia caused by Klebsiella species were collected from the hospital charts at the HELIOS Clinic, Witten/Herdecke University, Wuppertal, Germany, within the study period 2004 to 2014. An antibiogram was created from all of the study patients with pneumonia caused by Klebsiella species. Sensitivity and resistance profiles were performed for the different antibiotics that have been consistently used in the treatment of patients with pneumonia caused by Klebsiella species. All demographic, clinical, and laboratory data of all of the patients with pneumonia caused by Klebsiella species were collected from the patients' records.During the study period of January 1, 2004, to August 12, 2014, 149 patients were identified with community- and nosocomial-acquired pneumonia affected by Klebsiella species. These patients had a mean age of 70.6 ± 13 (107 [71.8%, 95% CI 64.6%-79%] men and 42 [28.2%, 95% CI 21%-35.4%] women). In all of the patients with pneumonia caused by Klebsiella species, there was resistance to ampicillin (P < 0.0001). Many patients with pneumonia caused by Klebsiella species (75.3%) also showed resistance to piperacillin (P < 0.0001). However, no patients with pneumonia caused by Klebsiella species showed resistance to imipenem or meropenem (P < 0.0001).Antibiotic resistance to the antibiotic class of carbapenem was not detected in patients with pneumonia caused by Klebsiella species.

  11. A one-step multiplex PCR to identify Klebsiella pneumoniae, Klebsiella variicola, and Klebsiella quasipneumoniae in the clinical routine.

    PubMed

    Fonseca, Erica Lourenço; Ramos, Nilceia da Veiga; Andrade, Bruno G Nascimento; Morais, Lena L C S; Marin, Michel F Abanto; Vicente, Ana Carolina P

    2017-04-01

    Klebsiella pneumoniae, Klebsiella variicola and Klebsiella quasipneumoniae are difficult to differentiate phenotypically, leading to misinterpretation of their infection prevalence. We propose a multiplex PCR for blaSHV, blaLEN and blaOKP and their flanking gene (deoR). Since this scheme focuses only on chromosomal genes, it will be feasible for Klebsiella identification in the clinical routine.

  12. Lung adenocarcinoma masquerading as refractory Klebsiella pneumoniae.

    PubMed

    McCartney, Clair; Moghadam, Afshin; Sriram, Krishna B

    2014-04-04

    We report the case of a middle-aged man where a diagnosis of Klebsiella pneumoniae obscured the underlying malignancy. The patient was hospitalised for management of a presumed refractory community-acquired pneumonia with radiological features of right lower lobe consolidation. Bronchoscopy did not identify an endobronchial lesion and washings grew K pneumoniae. CT-guided fine-needle aspirate samples did not detect any malignancy. However, despite appropriate antibiotic treatment there was no improvement in the patient's clinical condition. Consequently, a CT-guided lung core biopsy was performed to obtain more tissue for histopathology, which was diagnostic of primary lung adenocarcinoma. This case highlights the need to continue to investigate a patient who is not progressing as clinically appropriate to their original diagnosis.

  13. Vector promoters used in Klebsiella pneumoniae.

    PubMed

    Jiang, Xiao; Zhu, Chengqian; Lin, Jie; Li, Jingkang; Fu, Shuilin; Gong, Heng

    2016-09-01

    Much effort has been devoted to the metabolic engineering of Klebsiella pneumoniae; however, our knowledge of the actual expression level of promoters used in K. pneumoniae is limited. In this study, the expression levels of three promoters were compared systematically by using the lacZ reporter gene with different carbon sources in K. pneumoniae. The results showed that, although promoters PT5 and Ptac designed for Escherichia coli were functional, PT5 appeared more efficient and the induction/repression ratio of Ptac was decreased extremely in K. pneumoniae. The basal level of Ptac for lacZ expression reached 396.5 U/mg, which was 9.5-fold higher compared with PT5 in LB medium, indicating Ptac can be used as an efficient "constitutive" promoter as well as an efficient induced promoter in K. pneumoniae. In different carbon sources medium, a newly constructed endogenous constitutive Pbud proved to be a stable and weak promoter. On the basis of our data, a set of Pbud and Ptac promoters could meet the broad range (about 1,000 orders of magnitude) of gene expression needed for engineered K. pneumoniae in glycerol-based medium.

  14. Klebsiella pneumoniae FimK Promotes Virulence in Murine Pneumonia.

    PubMed

    Rosen, David A; Hilliard, Julia K; Tiemann, Kristin M; Todd, Elizabeth M; Morley, S Celeste; Hunstad, David A

    2016-02-15

    Klebsiella pneumoniae, a chief cause of nosocomial pneumonia, is a versatile and commonly multidrug-resistant human pathogen for which further insight into pathogenesis is needed. We show that the pilus regulatory gene fimK promotes the virulence of K. pneumoniae strain TOP52 in murine pneumonia. This contrasts with the attenuating effect of fimK on urinary tract virulence, illustrating that a single factor may exert opposing effects on pathogenesis in distinct host niches. Loss of fimK in TOP52 pneumonia was associated with diminished lung bacterial burden, limited innate responses within the lung, and improved host survival. FimK expression was shown to promote serum resistance, capsule production, and protection from phagocytosis by host immune cells. Finally, while the widely used K. pneumoniae model strain 43816 produces rapid dissemination and death in mice, TOP52 caused largely localized pneumonia with limited lethality, thereby providing an alternative tool for studying K. pneumoniae pathogenesis and control within the lung. © The Author 2015. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

  15. Klebsiella pneumoniae FimK Promotes Virulence in Murine Pneumonia

    PubMed Central

    Rosen, David A.; Hilliard, Julia K.; Tiemann, Kristin M.; Todd, Elizabeth M.; Morley, S. Celeste; Hunstad, David A.

    2016-01-01

    Klebsiella pneumoniae, a chief cause of nosocomial pneumonia, is a versatile and commonly multidrug-resistant human pathogen for which further insight into pathogenesis is needed. We show that the pilus regulatory gene fimK promotes the virulence of K. pneumoniae strain TOP52 in murine pneumonia. This contrasts with the attenuating effect of fimK on urinary tract virulence, illustrating that a single factor may exert opposing effects on pathogenesis in distinct host niches. Loss of fimK in TOP52 pneumonia was associated with diminished lung bacterial burden, limited innate responses within the lung, and improved host survival. FimK expression was shown to promote serum resistance, capsule production, and protection from phagocytosis by host immune cells. Finally, while the widely used K. pneumoniae model strain 43816 produces rapid dissemination and death in mice, TOP52 caused largely localized pneumonia with limited lethality, thereby providing an alternative tool for studying K. pneumoniae pathogenesis and control within the lung. PMID:26347570

  16. Perianal Abscess and Proctitis by Klebsiella pneumoniae.

    PubMed

    Jeong, Woo Shin; Choi, Sung Youn; Jeong, Eun Haeng; Bang, Ki Bae; Park, Seung Sik; Lee, Dae Sung; Park, Dong Il; Jung, Yoon Suk

    2015-01-01

    Klebsiella pneumoniae (K. pneumoniae) can at times cause invasive infections, especially in patients with diabetes mellitus and a history of alcohol abuse. A 61-year-old man with diabetes mellitus and a history of alcohol abuse presented with abdominal and anal pain for two weeks. After admission, he underwent sigmoidoscopy, which revealed multiple ulcerations with yellowish exudate in the rectum and sigmoid colon. The patient was treated with ciprofloxacin and metronidazole. After one week, follow up sigmoidoscopy was performed owing to sustained fever and diarrhea. The lesions were aggravated and seemed webbed in appearance because of damage to the rectal mucosa. Abdominal computed tomography and rectal magnetic resonance imaging were performed, and showed a perianal and perirectal abscess. The patient underwent laparoscopic sigmoid colostomy and perirectal abscess incision and drainage. Extended-spectrum beta-lactamase-producing K. pneumoniae was identified in pus culture. The antibiotics were switched to ertapenem. He improved after surgery and was discharged. K. pneumoniae can cause rapid invasive infection in patients with diabetes and a history of alcohol abuse. We report the first rare case of proctitis and perianal abscess caused by invasive K. pneumoniae infection.

  17. [Fatal pneumonia caused by carbapenem resistant Klebsiella pneumoniae].

    PubMed

    van Apeldoorn, Marjan; Lettinga, Kamilla; Bernards, Alexandra; Paltansing, Sunita; alNaiemi, Nashwan; Kalpoe, Jayant

    2010-01-01

    A 63-year-old Dutch man became colonized with a carbapenem resistant Klebsiella pneumoniae during a period of hospitalization in India. His recovery in the Netherlands was complicated by pneumonia due to this difficult-to-control multiresistant bacteria to which he eventually succumbed. Carbapenem resistance in Enterobacteriaceae, such as K. pneumoniae, is usually caused by carbapenemase (a betalactamase) production. Carbapenemase producing Enterobacteriaceae (CPE) are spreading throughout the world and cause difficult-to-treat infections that are associated with high mortality. This case report illustrates the clinical challenges associated with infection with these multiresistant Enterobacteriaceae. In the Netherlands, there are no guidelines for detection of CPE and carbapenemase production can frequently go undetected in clinical microbiology laboratories. As a consequence, adequate treatment of CPE infections and infection control measures to prevent the spread of CPE can be delayed. Expeditious development and implementation of existing Dutch draft guidelines for detection methods of CPE is therefore warranted.

  18. The diversity of Klebsiella pneumoniae surface polysaccharides

    PubMed Central

    Heinz, Eva; Wyres, Kelly L.; Ellington, Matthew J.; Kowarik, Michael; Holt, Kathryn E.; Thomson, Nicholas R.

    2016-01-01

    Klebsiella pneumoniae is considered an urgent health concern due to the emergence of multi-drug-resistant strains for which vaccination offers a potential remedy. Vaccines based on surface polysaccharides are highly promising but need to address the high diversity of surface-exposed polysaccharides, synthesized as O-antigens (lipopolysaccharide, LPS) and K-antigens (capsule polysaccharide, CPS), present in K. pneumoniae. We present a comprehensive and clinically relevant study of the diversity of O- and K-antigen biosynthesis gene clusters across a global collection of over 500 K. pneumoniae whole-genome sequences and the seroepidemiology of human isolates from different infection types. Our study defines the genetic diversity of O- and K-antigen biosynthesis cluster sequences across this collection, identifying sequences for known serotypes as well as identifying novel LPS and CPS gene clusters found in circulating contemporary isolates. Serotypes O1, O2 and O3 were most prevalent in our sample set, accounting for approximately 80 % of all infections. In contrast, K serotypes showed an order of magnitude higher diversity and differ among infection types. In addition we investigated a potential association of O or K serotypes with phylogenetic lineage, infection type and the presence of known virulence genes. K1 and K2 serotypes, which are associated with hypervirulent K. pneumoniae, were associated with a higher abundance of virulence genes and more diverse O serotypes compared to other common K serotypes. PMID:28348868

  19. [Clinical features and antimicrobial resistance of community-acquired pneumonia caused by Klebsiella pneumoniae in infants].

    PubMed

    He, Li-Yun; Wang, Ying-Jian; Li, Ji-Mei

    2012-11-01

    To study the clinical features and antimicrobial resistance of community-acquired pneumonia caused by Klebsiella pneumoniae in infants. The clinical data of 65 infants with community-acquired pneumonia caused by Klebsiella pneumoniae between 2007 and 2011 were retrospectively studied. Of the 65 infants, 37 cases (57%) were aged ≤3 months, 17 cases (26%) over 4 months, 7 cases (11%) over 7 months and 4 cases (6%) between 13 and 24 months. There were no significant differences in clinical manifestations and chest X-ray features between the infants with community-acquired pneumonia caused by Klebsiella pneumoniae and those with other bacterial pneumonia. Forty strains (62%) of ESBLs-producing Klebsiella pneumoniae were detected. Klebsiella pneumoniae was 100% sensitive to imipenem, meropenem and amikacin but resistant to penicillins and cephalosporins. The resistance rates of ESBLs-producing strains to penicillins, cephalosporins, amoxicillin/clavulanic acid, ampicillin/sulbactam, compound sulfamethoxazole, gentamycin, ciprofloxacin and aztreonam were significantly higher than for non-ESBLs-producing strains. ESBLs-producing strains also showed multiple-drug resistance. Community-acquired pneumonia caused by Klebsiella pneumoniae is common in infants aged ≤3 months. ESBLs-producing strains are prevalent in community-acquired pneumonia caused by Klebsiella pneumoniae and demonstrate both high rates of drug resistance and multiple-drug resistance.

  20. Klebsiella pneumoniae survives within macrophages by avoiding delivery to lysosomes.

    PubMed

    Cano, Victoria; March, Catalina; Insua, Jose Luis; Aguiló, Nacho; Llobet, Enrique; Moranta, David; Regueiro, Verónica; Brennan, Gerard P; Millán-Lou, Maria Isabel; Martín, Carlos; Garmendia, Junkal; Bengoechea, José A

    2015-11-01

    Klebsiella pneumoniae is an important cause of community-acquired and nosocomial pneumonia. Evidence indicates that Klebsiella might be able to persist intracellularly within a vacuolar compartment. This study was designed to investigate the interaction between Klebsiella and macrophages. Engulfment of K. pneumoniae was dependent on host cytoskeleton, cell plasma membrane lipid rafts and the activation of phosphoinositide 3-kinase (PI3K). Microscopy studies revealed that K. pneumoniae resides within a vacuolar compartment, the Klebsiella-containing vacuole (KCV), which traffics within vacuoles associated with the endocytic pathway. In contrast to UV-killed bacteria, the majority of live bacteria did not co-localize with markers of the lysosomal compartment. Our data suggest that K. pneumoniae triggers a programmed cell death in macrophages displaying features of apoptosis. Our efforts to identify the mechanism(s) whereby K. pneumoniae prevents the fusion of the lysosomes to the KCV uncovered the central role of the PI3K-Akt-Rab14 axis to control the phagosome maturation. Our data revealed that the capsule is dispensable for Klebsiella intracellular survival if bacteria were not opsonized. Furthermore, the environment found by Klebsiella within the KCV triggered the down-regulation of the expression of cps. Altogether, this study proves evidence that K. pneumoniae survives killing by macrophages by manipulating phagosome maturation that may contribute to Klebsiella pathogenesis.

  1. The capsular network of Klebsiella pneumoniae.

    PubMed

    Cassone, A; Garaci, E

    1977-06-01

    Attempts at improving chemical fixation for electron-microscopic observation of the capsule of Klebsiella pneumoniae were made. The capsule was preserved by using alcian blue - lanthanum and tris-(1-aziridinyl) phosphine oxide (TAPO) - aldehyde - osmium procedures. Despite the different retention of the overall capsular material and minor variations in morphological details, in both cases the interpretation of ultrastructural patterns suggested that the capsule be composed of a meshed network of thin polysaccharide fibrils radiating from the cell wall. This organization is in keeping with all recognized chemical properties of bacterial polysaccharide capsules or, at least, does not contradict them. Moreover, an effective preservation of bacterial structures other than capsule has been obtained, mostly in specimens fixed by the TAPO-aldehyde-osmium method, a fact which gives further reliability to the technical approach used for capsule visualization.

  2. Mapping the Evolution of Hypervirulent Klebsiella pneumoniae

    PubMed Central

    Roe, Chandler C.; Stegger, Marc; Stahlhut, Steen G.; Hansen, Dennis S.; Engelthaler, David M.; Andersen, Paal S.; Driebe, Elizabeth M.; Keim, Paul; Krogfelt, Karen A.

    2015-01-01

    ABSTRACT Highly invasive, community-acquired Klebsiella pneumoniae infections have recently emerged, resulting in pyogenic liver abscesses. These infections are caused by hypervirulent K. pneumoniae (hvKP) isolates primarily of capsule serotype K1 or K2. Hypervirulent K1 isolates belong to clonal complex 23 (CC23), indicating that this clonal lineage has a specific genetic background conferring hypervirulence. Here, we apply whole-genome sequencing to a collection of K. pneumoniae isolates to characterize the phylogenetic background of hvKP isolates with an emphasis on CC23. Most of the hvKP isolates belonged to CC23 and grouped into a distinct monophyletic clade, revealing that CC23 is a unique clonal lineage, clearly distinct from nonhypervirulent strains. Separate phylogenetic analyses of the CC23 isolates indicated that the CC23 lineage evolved recently by clonal expansion from a single common ancestor. Limited grouping according to geographical origin was observed, suggesting that CC23 has spread globally through multiple international transmissions. Conversely, hypervirulent K2 strains clustered in genetically unrelated groups. Strikingly, homologues of a large virulence plasmid were detected in all hvKP clonal lineages, indicating a key role in K. pneumoniae hypervirulence. The plasmid encodes two siderophores, aerobactin and salmochelin, and RmpA (regulator of the mucoid phenotype); all these factors were found to be restricted to hvKP isolates. Genomic comparisons revealed additional factors specifically associated with CC23. These included a distinct variant of a genomic island encoding yersiniabactin, colibactin, and microcin E492. Furthermore, additional novel genomic regions unique to CC23 were revealed which may also be involved in the increased virulence of this important clonal lineage. PMID:26199326

  3. Draft Genome Sequence of Klebsiella pneumoniae subsp. pneumoniae ATCC 9621.

    PubMed

    Poehlein, Anja; Najdenski, Hristo; Simeonova, Diliana D

    2017-03-23

    We present here the 5.561-Mbp assembled draft genome sequence of Klebsiella pneumoniae subsp. pneumoniae ATCC 9621, a phosphite- and organophosphonate-assimilating Gammaproteobacterium. The genome harbors 5,179 predicted protein-coding genes.

  4. Receptor for Advanced Glycation End Products (RAGE) Serves a Protective Role during Klebsiella pneumoniae - Induced Pneumonia.

    PubMed

    Achouiti, Ahmed; de Vos, Alex F; van 't Veer, Cornelis; Florquin, Sandrine; Tanck, Michael W; Nawroth, Peter P; Bierhaus, Angelika; van der Poll, Tom; van Zoelen, Marieke A D

    2016-01-01

    Klebsiella species is the second most commonly isolated gram-negative organism in sepsis and a frequent causative pathogen in pneumonia. The receptor for advanced glycation end products (RAGE) is expressed on different cell types and plays a key role in diverse inflammatory responses. We here aimed to investigate the role of RAGE in the host response to Klebsiella (K.) pneumoniae pneumonia and intransally inoculated rage gene deficient (RAGE-/-) and normal wild-type (Wt) mice with K. pneumoniae. Klebsiella pneumonia resulted in an increased pulmonary expression of RAGE. Furthermore, the high-affinity RAGE ligand high mobility group box-1 was upregulated during K. pneumoniae pneumonia. RAGE deficiency impaired host defense as reflected by a worsened survival, increased bacterial outgrowth and dissemination in RAGE-/- mice. RAGE-/- neutrophils showed a diminished phagocytosing capacity of live K. pneumoniae in vitro. Relative to Wt mice, RAGE-/- mice demonstrated similar lung inflammation, and slightly elevated-if any-cytokine and chemokine levels and unchanged hepatocellular injury. In addition, RAGE-/- mice displayed an unaltered response to intranasally instilled Klebsiella lipopolysaccharide (LPS) with respect to pulmonary cell recruitment and local release of cytokines and chemokines. These data suggest that (endogenous) RAGE protects against K. pneumoniae pneumonia. Also, they demonstrate that RAGE contributes to an effective antibacterial defense during K. pneumoniae pneumonia, at least partly via its participation in the phagocytic properties of professional granulocytes. Additionally, our results indicate that RAGE is not essential for the induction of a local and systemic inflammatory response to either intact Klebsiella or Klebsiella LPS.

  5. Whole-Genome Sequencing of Human Clinical Klebsiella pneumoniae Isolates Reveals Misidentification and Misunderstandings of Klebsiella pneumoniae, Klebsiella variicola, and Klebsiella quasipneumoniae

    PubMed Central

    Linson, Sarah E.; Ojeda Saavedra, Matthew; Cantu, Concepcion; Davis, James J.; Brettin, Thomas; Olsen, Randall J.

    2017-01-01

    ABSTRACT Klebsiella pneumoniae is a major threat to public health, causing significant morbidity and mortality worldwide. The emergence of highly drug-resistant strains is particularly concerning. There has been a recognition and division of Klebsiella pneumoniae into three distinct phylogenetic groups: Klebsiella pneumoniae, Klebsiella variicola, and Klebsiella quasipneumoniae. K. variicola and K. quasipneumoniae have often been described as opportunistic pathogens that have less virulence in humans than K. pneumoniae does. We recently sequenced the genomes of 1,777 extended-spectrum-beta-lactamase (ESBL)-producing K. pneumoniae isolates recovered from human infections and discovered that 28 strains were phylogenetically related to K. variicola and K. quasipneumoniae. Whole-genome sequencing of 95 additional non-ESBL-producing K. pneumoniae isolates recovered from patients found 12 K. quasipneumoniae strains. Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) analysis initially identified all patient isolates as K. pneumoniae, suggesting a potential pitfall in conventional clinical microbiology laboratory identification methods. Whole-genome sequence analysis revealed extensive sharing of core gene content and plasmid replicons among the Klebsiella species. For the first time, strains of both K. variicola and K. quasipneumoniae were found to carry the Klebsiella pneumoniae carbapenemase (KPC) gene, while another K. variicola strain was found to carry the New Delhi metallo-beta-lactamase 1 (NDM-1) gene. K. variicola and K. quasipneumoniae infections were not less virulent than K. pneumoniae infections, as assessed by in-hospital mortality and infection type. We also discovered evidence of homologous recombination in one K. variicola strain, as well as one strain from a novel Klebsiella species, which challenge the current understanding of interrelationships between clades of Klebsiella. IMPORTANCE Klebsiella

  6. [A study on carbapenem resistance in klebsiella pneumoniae].

    PubMed

    Guan, Hong; Cao, Xia; Chen, Rui; Zhou, Tao; Huang, Xing-Long; Xu, Xin; Pei, Xiao-Fang

    2013-03-01

    To investigate the molecular mechanisms of reduced carbapenem susceptibility in Klebsiella pneumonia. One reduced carbapenem susceptible Klebsiella pneumonia clinical isolate was investigated. Kirby-Bauer disc test was applied to determine the antibiotic susceptibility of the isolate. Modified Hodge Test and EDTA-disk synergy test were used to confirm whether this Klebsiella pneumonia strain could produce metallo-beta-lactamase. The genotype of the beta-lactamase was confirmed by PCR and DNA sequence analysis. Plasmid DNA preparations and conjugation experiment were used to determine the location of the resistant gene. Antibacterial circle of imipenem, meropenem for Klebsiella pneumonia isolate were 16 cm and 17 cm implied that the isolated strain producing carbapenemas. Modified Hodge Test and EDTA-disk synergy test confirmed that this Klebsiella pneumonia isolate produced metallo-beta-lactamase. IMP-4 gene was amplified by PCR and confirmed with sequence analysis. A reduced carbapenem susceptibility in obtained conjugants was observed when evaluated with Kirby-Bauer disc test and conjugation experiment also revealed that blalMP-4 were carried on one plasmid with a size of approximately 73 000 bp. Production of plasmid-mediated metallo-beta lactamase IMP-4 might lead to the reduced susceptibility of Klebsiella pneumonia spp. to carbapenems.

  7. Nosocomial Outbreak of Klebsiella pneumoniae Carbapenemase-Producing Klebsiella oxytoca in Austria

    PubMed Central

    Hoenigl, Martin; Valentin, Thomas; Zarfel, Gernot; Wuerstl, Benjamin; Leitner, Eva; Salzer, Helmut J. F.; Posch, Josefa; Krause, Robert

    2012-01-01

    To date, no outbreak of carbapenemase-producing bacteria has been reported for Austria. While outbreaks of Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae have been increasingly reported, no outbreak caused by KPC-producing Klebsiella oxytoca has been described yet, to the best of our knowledge. We report an outbreak of KPC-producing K. oxytoca. In 5 months, 31 KPC-producing Klebsiella oxytoca strains were isolated from five patients. All patients were admitted to the same medical intensive care unit in Austria. PMID:22290949

  8. Nosocomial outbreak of Klebsiella pneumoniae carbapenemase-producing Klebsiella oxytoca in Austria.

    PubMed

    Hoenigl, Martin; Valentin, Thomas; Zarfel, Gernot; Wuerstl, Benjamin; Leitner, Eva; Salzer, Helmut J F; Posch, Josefa; Krause, Robert; Grisold, Andrea J

    2012-04-01

    To date, no outbreak of carbapenemase-producing bacteria has been reported for Austria. While outbreaks of Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae have been increasingly reported, no outbreak caused by KPC-producing Klebsiella oxytoca has been described yet, to the best of our knowledge. We report an outbreak of KPC-producing K. oxytoca. In 5 months, 31 KPC-producing Klebsiella oxytoca strains were isolated from five patients. All patients were admitted to the same medical intensive care unit in Austria.

  9. Antimicrobial resistance temporal trend of Klebsiella pneumoniae isolated from blood.

    PubMed

    Gavriliu, L C; Benea, O E; Benea, S

    2016-01-01

    Background. According to the European Antimicrobial Resistance Surveillance Network, Romania reports an increasing number of resistant Klebsiella pneumoniae strains from invasive infections every year. Material and Method. We analyzed the antimicrobial susceptibility of Klebsiella pneumoniae strains isolated from blood in 2010 and 2015 in "Matei Bals" National Institute of Infectious Diseases, in order to identify any significant changes in the last five years. Results. We identified 18 strains in 2010 and 37 strains in 2015. Although the resistance to aminopenicillin-betalactamase inhibitors association, piperacillin-tazobactam, third generation cephalosporins, fluoroquinolones, gentamicin, amikacin and the combined resistance decreased between these two time frames, this evolution was statistically non-significant. The same was noticed for the increased resistance rates to carbapenems. Conclusions. Antimicrobial resistance of Klebsiella pneumoniae may become a major problem for the public health and the hospital-acquired infections control. Therefore, it needs further monitoring and efforts must be made in order to limit the increase of the resistance.

  10. Evidence for a nicotinamide nucleotide transhydrogenase in Klebsiella pneumoniae.

    PubMed

    Fristedt, U; Rydström, J; Persson, B

    1994-02-15

    Bacterial membranes from Klebsiella pneumoniae were investigated for the presence of a nicotinamide nucleotide transhydrogenase activity. Inverted membrane vesicles derived from these cells catalyzed a reduction of NAD+ or 3-acetylpyridine-NAD+ by NADPH, which showed a maximal activity of about 260 nmoles/minute per milligram protein at pH 7-8. In the presence of a protonic uncoupler the specific activity was stimulated about two-fold in this pH range. The presence of detergents did not further increase the specific activity of enzyme. The Klebsiella pneumoniae transhydrogenase activity was sensitive to phenylarsine oxide and palmityl-Coenzyme A, both of which are agents known to inhibit the mammalian enzyme. The Ki-value for palmityl-Coenzyme A with respect to NADPH was about 1.25 microM. Antibodies raised against beef heart transhydrogenase crossreacted with a 54 kD protein in the Klebsiella pneumonia membrane.

  11. First report of KPC-producing Klebsiella pneumoniae in Croatia.

    PubMed

    Bedenić, Branka; Mazzariol, Annarita; Plečko, Vanda; Bošnjak, Zrinka; Barl, Petra; Vraneš, Jasmina; Cornaglia, Giuseppe

    2012-08-01

    In February 2011, a 78-year-old male patient was admitted to Clinical Hospital Center Zagreb with subdural haematoma. Klebsiella pneumoniae with reduced susceptibility to carbapenems was isolated. PCR revealed the presence of bla(KPC), bla(TEM), and bla(SHV) genes. Sequencing of bla(KPC) gene identified K. pneumoniae carbapenemase (KPC)-2 beta-lactamase. The strain belonged to ST37 clone by multilocus sequence typing. Infection control efforts limited the spread of KPC-producing clone of K. pneumoniae in our hospital so far. To our knowledge, this is the first report of a KPC-producing K. pneumoniae in Croatia.

  12. Receptor for Advanced Glycation End Products (RAGE) Serves a Protective Role during Klebsiella pneumoniae - Induced Pneumonia

    PubMed Central

    Achouiti, Ahmed; de Vos, Alex F.; van ‘t Veer, Cornelis; Florquin, Sandrine; Tanck, Michael W.; Nawroth, Peter P.; Bierhaus, Angelika; van der Poll, Tom; van Zoelen, Marieke A. D.

    2016-01-01

    Klebsiella species is the second most commonly isolated gram-negative organism in sepsis and a frequent causative pathogen in pneumonia. The receptor for advanced glycation end products (RAGE) is expressed on different cell types and plays a key role in diverse inflammatory responses. We here aimed to investigate the role of RAGE in the host response to Klebsiella (K.) pneumoniae pneumonia and intransally inoculated rage gene deficient (RAGE-/-) and normal wild-type (Wt) mice with K. pneumoniae. Klebsiella pneumonia resulted in an increased pulmonary expression of RAGE. Furthermore, the high-affinity RAGE ligand high mobility group box-1 was upregulated during K. pneumoniae pneumonia. RAGE deficiency impaired host defense as reflected by a worsened survival, increased bacterial outgrowth and dissemination in RAGE-/- mice. RAGE-/- neutrophils showed a diminished phagocytosing capacity of live K. pneumoniae in vitro. Relative to Wt mice, RAGE-/- mice demonstrated similar lung inflammation, and slightly elevated—if any—cytokine and chemokine levels and unchanged hepatocellular injury. In addition, RAGE-/- mice displayed an unaltered response to intranasally instilled Klebsiella lipopolysaccharide (LPS) with respect to pulmonary cell recruitment and local release of cytokines and chemokines. These data suggest that (endogenous) RAGE protects against K. pneumoniae pneumonia. Also, they demonstrate that RAGE contributes to an effective antibacterial defense during K. pneumoniae pneumonia, at least partly via its participation in the phagocytic properties of professional granulocytes. Additionally, our results indicate that RAGE is not essential for the induction of a local and systemic inflammatory response to either intact Klebsiella or Klebsiella LPS. PMID:26824892

  13. Acute renal failure caused by Klebsiella pneumoniae pyelonephritis.

    PubMed

    Creyghton, W M; Lobatto, S; Weening, J J

    2001-11-01

    We report a 34-year-old male patient without prior medical history who presented with acute renal failure due to acute bacterial pyelonephritis. Both blood and urine cultures grew Klebsiella pneumoniae. Although a kidney biopsy revealed extensive necrosis and no viable glomeruli, renal function recovered to near normal after intermittent hemodialysis and antibiotic therapy. We believe that it is important to include this entity in the differential diagnosis of acute renal failure since proper diagnosis and treatment is essential for recovery of renal function. Furthermore, we would like to draw attention to Klebsiella pneumoniae as an important potential pathogen in such cases, in addition to Escherichia coli.

  14. Pneumonia Caused by Klebsiella spp. in 46 Horses.

    PubMed

    Estell, K E; Young, A; Kozikowski, T; Swain, E A; Byrne, B A; Reilly, C M; Kass, P H; Aleman, M

    2016-01-01

    Klebsiella spp. are implicated as a common cause of bacterial pneumonia in horses, but few reports describe clinical presentation and disease progression. To describe the signalment, clinicopathologic data, radiographic and ultrasonographic findings, antimicrobial susceptibility, outcome, and pathologic lesions associated with Klebsiella spp. pneumonia in horses. Forty-six horses from which Klebsiella spp. was isolated from the lower respiratory tract. Retrospective study. Medical records from 1993 to 2013 at the William R. Pritchard Veterinary Medical Teaching Hospital, University of California, Davis were reviewed. Exact logistic regression was performed to determine if any variables were associated with survival to hospital discharge. Survival in horses <1 year old was 73%. Overall survival in adults was 63%. For adults in which Klebsiella pneumoniae was the primary isolate, survival was 52%. Mechanical ventilation preceded development of pneumonia in 11 horses. Complications occurred in 25/46 horses, with thrombophlebitis and laminitis occurring most frequently. Multi-drug resistance was found in 47% of bacterial isolates. Variables that significantly impacted survival included hemorrhagic nasal discharge, laminitis, and thoracic radiographs with a sharp demarcation between marked caudal pulmonary alveolar infiltration and more normal-appearing caudodorsal lung. Klebsiella spp. should be considered as a differential diagnosis for horses presenting with hemorrhagic pneumonia and for horses developing pneumonia after mechanical ventilation. Multi-drug resistance is common. Prognosis for survival generally is fair, but is guarded for adult horses in which K. pneumoniae is isolated as the primary organism. Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  15. Chronic pneumonia due to Klebsiella oxytoca mimicking pulmonary tuberculosis.

    PubMed

    Gera, Kamal; Roshan, Rahul; Varma-Basil, Mandira; Shah, Ashok

    2015-01-01

    Klebsiella species infrequently cause acute community acquired pneumonia (CAP). The chronic form of the disease caused by K. pneumoniae (Friedlander's bacillus) was occasionally seen in the pre-antibiotic era. K. oxytoca is a singularly uncommon cause of CAP. The chronic form of the disease caused by K. oxytoca has been documented only once before. A 50-year-old immunocompetent male smoker presented with haemoptysis for 12 months. Imaging demonstrated a cavitary lesion in the right upper lobe with emphysematous changes. Sputum stains and cultures for Mycobacterium tuberculosis were negative. However, three sputum samples for aerobic culture as well as bronchial aspirate cultured pure growth of K. oxytoca. A diagnosis of chronic pneumonia due to K. oxytoca was established and with appropriate therapy, the patient was largely asymptomatic. The remarkable clinical and radiological similarity to pulmonary tuberculosis can result in patients with chronic Klebsiella pneumonia erroneously receiving anti-tuberculous therapy.

  16. Order of genes near nif in Klebsiella pneumoniae.

    PubMed Central

    MacNeil, D; Supiano, M A; Brill, W J

    1979-01-01

    Analysis of strains with deletions of all or part of nif have ordered the Klebsiella pneumoniae genetic loci as thi rbt dal udk gnd rfb has nif shiA. The his-nif plasmids pRD1 and pTM4010 contain the genes gnd rfb his nif shiA. PMID:378930

  17. Implantable cardioverter defibrillator pocket infection caused by Klebsiella pneumonia.

    PubMed

    Ertas, F; Acet, H; Kaya, H; Kayan, F; Soydinc, S

    2012-09-01

    Like any other foreign bodies, implanted cardiac devices can become infected. Staphylococcus aureus and coagulase-negative Staphilococci are the most common causes of infections of pacemaker and defibrillator systems. In this case an implantable cardioverter defibrillator pocket infection caused by an extremely rare microorganism, Klebsiella pneumonia, is presented.

  18. Klebsiella pneumoniae in Gastrointestinal Tract and Pyogenic Liver Abscess

    PubMed Central

    Lin, Yi-Tsung; Lin, Jung-Chung; Chen, Te-Li; Yeh, Kuo-Ming; Chang, Feng-Yee; Chuang, Han-Chuan; Wu, Hau-Shin; Tseng, Chih-Peng; Siu, L. Kristopher

    2012-01-01

    To determine the role of gastrointestinal carriage in Klebsiella pneumoniae liver abscess, we studied 43 patients. Bacterial isolates from liver and fecal samples from 10 patients with this condition and 7 healthy carriers showed identical serotypes and genotypes with the same virulence. This finding indicated that gastrointestinal carriage is a predisposing factor for liver abscess. PMID:22840473

  19. Draft Genome Sequence of Klebsiella pneumoniae subsp. pneumoniae ATCC 9621

    PubMed Central

    Najdenski, Hristo

    2017-01-01

    ABSTRACT We present here the 5.561-Mbp assembled draft genome sequence of Klebsiella pneumoniae subsp. pneumoniae ATCC 9621, a phosphite- and organophosphonate-assimilating Gammaproteobacterium. The genome harbors 5,179 predicted protein-coding genes. PMID:28336608

  20. Clinical and pulmonary thin-section CT findings in acute Klebsiella pneumoniae pneumonia.

    PubMed

    Okada, Fumito; Ando, Yumiko; Honda, Koichi; Nakayama, Tomoko; Kiyonaga, Maki; Ono, Asami; Tanoue, Shuichi; Maeda, Toru; Mori, Hiromu

    2009-04-01

    The aim of this study was to assess the clinical and pulmonary thin-section CT findings in patients with acute Klebsiella pneumoniae pneumonia. We retrospectively evaluated thin-section CT examinations performed between January 1991 and December 2007 from 962 patients with acute Klebsiella pneumoniae pneumonia. Seven hundred and sixty-four cases with concurrent infectious diseases were excluded. Thus, our study group comprised 198 patients (118 male, 80 female; age range 18-97 years, mean age 61.5). Underlying diseases and clinical findings were assessed. Parenchymal abnormalities were evaluated along with the presence of enlarged lymph nodes and pleural effusion. CT findings in patients with acute Klebsiella pneumoniae pneumonia consisted mainly of ground-glass attenuation (100%), consolidation (91.4%), and intralobular reticular opacity (85.9%), which were found in the periphery (96%) of both sides of the lungs (72.2%) and were often associated with pleural effusion (53%). The underlying conditions in patients with Klebsiella pneumoniae pneumonia were alcoholism or smoking habit.

  1. Detection of metallo-β-lactamase genes in clinically isolated Klebsiella pneumoniae and Klebsiella oxytoca.

    PubMed

    Tateno, Hidetsugu; Yasuhara, Tsutomu; Sugano, Emi; Tahara, Sachiko; Ugajin, Kazuhisa; Fukuchi, Kunihiko

    2014-12-01

    We detected and characterized metallo-β-lactamase genes (blaIMP-1 and blaIMP-11) in third generation cephalosporin-resistant Klebsiella pneumoniae and Klebsiella oxytoca isolated at Showa University Hospital between January 1, 2011 and December 31, 2012. The cephalosporin-resistant K pneumoniae strains were frequently isolated from the urine, while one strain of K. pneumoniae, which was resistant to carbapenem, was isolated from the stool. We analyzed the phenotypes and genotypes of the metallo-β-lactamase genes from the 16 strains of cephalosporin-resistant-K pneumoniae and 6 strains of -K. oxytoca isolated from the same ward. The minimum inhibitory concentrations of imipenem were below 4 µg/ml in 21 out of the 22 isolated strains. The double disc synergy test using ceftazidime and sodium mercaptoacetic acid revealed enlargements in the inhibitory zones of 14 of the 16 strains of K. pneumoniae and all 6 strains of K. oxytoca. Metallo-β-lactamase genes were detected in all of the tested strains, with blaIMP-1 in 3 K. pneumoniae and 1 K. oxytoca, blaIMP-11 in 13 K pneumoniae and 4 K. oxytoca, and both blaIMP-1 and blaIMP-11 in one K. oxytoca. Our results indicate that third generation cephalosporin-resistant and imipenem-susceptible K. pneumoniae and K. oxytoca possess the metallo-β-lactamase gene. The active surveillance of metallo-β-lactamase genes should be performed in clinical laboratories. (Original).

  2. Klebsiella pneumoniae necrotizing fasciitis in a Latin American male.

    PubMed

    Persichino, Jon; Tran, Richard; Sutjita, Made; Kim, Daniel

    2012-11-01

    Necrotizing fasciitis, caused by Klebsiella pneumoniae, is a rare and life-threatening bacterial infection. Most documented cases have been reported from Asia, particularly associated with diabetes mellitus. The prevalence of this infection in the USA is rare, especially among persons of non-Asian descent and those without travel to Asia. We report a case of disseminated necrotizing fasciitis, caused by K. pneumoniae, in a Latin American male with diabetes mellitus. Given our review of the literature, this is the only case report, to our knowledge, of a Latin American patient with Klebsiella necrotizing fasciitis in the USA. This case may reflect the geographical spread and emergence of K. pneumoniae infection in the USA. Clinicians need to be aware of the possible relationship between this organism and necrotizing fasciitis in persons of Latin American descent with diabetes mellitus.

  3. The K1 beta-lactamase of Klebsiella pneumoniae.

    PubMed Central

    Joris, B; De Meester, F; Galleni, M; Frère, J M; Van Beeumen, J

    1987-01-01

    beta-Lactamase K1 was purified from Klebsiella pneumoniae SC10436. It is very similar to the enzyme produced by Klebsiella aerogenes 1082E and described by Emanuel, Gagnon & Waley [Biochem. J. (1986) 234, 343-347]. An active-site peptide was isolated after labelling of the enzyme with tritiated beta-iodopenicillanate. A cysteine residue was found just before the active-site serine residue. This result could explain the properties of the enzyme after modification by thiol-blocking reagents. The sequence of the active-site peptide clearly established the enzyme as a class A beta-lactamase. PMID:3307765

  4. Epidemiological typing of Klebsiella pneumoniae by pyrolysis mass spectrometry.

    PubMed

    Jackson, R M; Heginbothom, M L; Magee, J T

    1997-01-01

    Thirteen isolates of ceftazidime-resistant Klebsiella pneumoniae from a suspected cross-infection outbreak involving patients on an intensive care unit and a haematology ward were examined in pyrolysis-mass spectrometry (Py-MS), along with eight concurrent non-outbreak-associated clinical isolates of klebsiellae as controls. Py-MS showed tight clustering of the suspected outbreak isolates, suggesting cross-infection with a single strain. Non-outbreak isolates were clearly distinct from one another and from the outbreak strain. The results confirm that Py-MS is a powerful tool for rapid strain comparison in investigations of cross-infection incidents.

  5. Klebsiella pneumoniae triggers a cytotoxic effect on airway epithelial cells

    PubMed Central

    2009-01-01

    Background Klebsiella pneumoniae is a capsulated Gram negative bacterial pathogen and a frequent cause of nosocomial infections. Despite its clinical relevance, little is known about the features of the interaction between K. pneumoniae and lung epithelial cells on a cellular level, neither about the role of capsule polysaccharide, one of its best characterised virulence factors, in this interaction. Results The interaction between Klebsiella pneumoniae and cultured airway epithelial cells was analysed. K. pneumoniae infection triggered cytotoxicity, evident by cell rounding and detachment from the substrate. This effect required the presence of live bacteria and of capsule polysaccharide, since it was observed with isolates expressing different amounts of capsule and/or different serotypes but not with non-capsulated bacteria. Cytotoxicity was analysed by lactate dehydrogenase and formazan measurements, ethidium bromide uptake and analysis of DNA integrity, obtaining consistent and complementary results. Moreover, cytotoxicity of non-capsulated strains was restored by addition of purified capsule during infection. While a non-capsulated strain was avirulent in a mouse infection model, capsulated K. pneumoniae isolates displayed different degrees of virulence. Conclusion Our observations allocate a novel role to K. pneumoniae capsule in promotion of cytotoxicity. Although this effect is likely to be associated with virulence, strains expressing different capsule levels were not equally virulent. This fact suggests the existence of other bacterial requirements for virulence, together with capsule polysaccharide. PMID:19650888

  6. Draft Genome Sequence of Klebsiella pneumoniae Strain AS Isolated from Zhejiang Provincial Hospital of TCM, China

    PubMed Central

    Yang, Xue-Jing; Wang, Sai; Hou, Jia-Hui

    2016-01-01

    Klebsiella pneumoniae is a Gram-negative, nonmotile, encapsulated, lactose-fermenting, facultative anaerobic, rod-shaped bacterium. Here we present draft genome assemblies of Klebsiella pneumoniae AS, which was isolated in China. The genomic information will provide a better understanding of the physiology, adaptation, and evolution of K. pneumoniae. PMID:27660770

  7. Correlation between antimicrobial resistance and virulence in Klebsiella pneumoniae.

    PubMed

    Hennequin, C; Robin, F

    2016-03-01

    Klebsiella pneumoniae is responsible for a wide range of infections, including urinary tract infections, pneumonia, bacteremia, and liver abscesses. In addition to susceptible clinical isolates involved in nosocomial infections, multidrug-resistant (MDR) and hypervirulent (hvKP) strains have evolved separately in distinct clonal groups. The rapid geographic spread of these isolates is of particular concern. However, we still know little about the virulence of K. pneumoniae except for hvKP, whose secrets are beginning to be revealed. The treatment of K. pneumoniae infections is threatened by the emergence of antimicrobial resistance. The dissemination of resistance is associated with genetic mobile elements, such as plasmids that may also carry virulence determinants. A proficient pathogen should be virulent, resistant to antibiotics, and epidemic. However, the interplay between resistance and virulence is poorly understood. Here, we review current knowledge on the topic.

  8. Antibiotic Resistance Related to Biofilm Formation in Klebsiella pneumoniae

    PubMed Central

    Vuotto, Claudia; Longo, Francesca; Balice, Maria Pia; Donelli, Gianfranco; Varaldo, Pietro E.

    2014-01-01

    The Gram-negative opportunistic pathogen, Klebsiella pneumoniae, is responsible for causing a spectrum of community-acquired and nosocomial infections and typically infects patients with indwelling medical devices, especially urinary catheters, on which this microorganism is able to grow as a biofilm. The increasingly frequent acquisition of antibiotic resistance by K. pneumoniae strains has given rise to a global spread of this multidrug-resistant pathogen, mostly at the hospital level. This scenario is exacerbated when it is noted that intrinsic resistance to antimicrobial agents dramatically increases when K. pneumoniae strains grow as a biofilm. This review will summarize the findings about the antibiotic resistance related to biofilm formation in K. pneumoniae. PMID:25438022

  9. Klebsiella pneumoniae pharyngitis mimicking malignancy: a diagnostic dilemma.

    PubMed

    Yeh, C-F; Li, W-Y; Hsu, Y-B

    2014-12-01

    Acute pharyngitis is a common disease. However, acute pharyngitis caused by Klebsiella pneumoniae with a gross appearance mimicking hypopharyngeal malignancy has never previously been reported. We report the case of a 57-year-old man with a right hypopharyngeal tumor which was disclosed by fiberoptic laryngoscopy and computed tomography scan. However, both the frozen and final pathologies showed no evidence of malignant cells, and a bacterial culture revealed the growth of K. pneumoniae. The hypopharyngeal lesion completely regressed after 2 weeks of antibiotic treatment. Clinicians should perform biopsy along with tissue culture for tumor-like lesions because infectious agents can lead to lesions with malignancy-like appearance.

  10. A Second Outer-Core Region in Klebsiella pneumoniae Lipopolysaccharide†

    PubMed Central

    Regué, Miguel; Izquierdo, Luis; Fresno, Sandra; Piqué, Núria; Corsaro, Maria Michela; Naldi, Teresa; De Castro, Cristina; Waidelich, Dietmar; Merino, Susana; Tomás, Juan M.

    2005-01-01

    Up to now only one major type of core oligosaccharide has been found in the lipopolysaccharide of all Klebsiella pneumoniae strains analyzed. Applying a different screening approach, we identified a novel Klebsiella pneumoniae core (type 2). Both Klebsiella core types share the same inner core and the outer-core-proximal disaccharide, GlcN-(1,4)-GalA, but they differ in the GlcN substituents. In core type 2, the GlcpN residue is substituted at the O-4 position by the disaccharide β-Glcp(1-6)-α-Glcp(1, while in core type 1 the GlcpN residue is substituted at the O-6 position by either the disaccharide α-Hep(1-4)-α-Kdo(2 or a Kdo residue (Kdo is 3-deoxy-d-manno-octulosonic acid). This difference correlates with the presence of a three-gene region in the corresponding core biosynthetic clusters. Engineering of both core types by interchanging this specific region allowed studying the effect on virulence. The replacement of Klebsiella core type 1 in a highly type 2 virulent strain (52145) induces lower virulence than core type 2 in a murine infection model. PMID:15937181

  11. Complete Genome Sequence of Klebsiella pneumoniae YH43

    PubMed Central

    Ogura, Yoshitoshi; Hayashi, Tetsuya; Mizunoe, Yoshimitsu

    2016-01-01

    We report here the complete genome sequence of Klebsiella pneumoniae strain YH43, isolated from sweet potato. The genome consists of a single circular chromosome of 5,520,319 bp in length. It carries 8 copies of rRNA operons, 86 tRNA genes, 5,154 protein-coding genes, and the nif gene cluster for nitrogen fixation. PMID:27081127

  12. Draft Genome Sequence of Klebsiella pneumoniae AWD5

    PubMed Central

    Rajkumari, Jina; Singha, L. Paikhomba

    2017-01-01

    ABSTRACT Here, we report the draft genome sequence of Klebsiella pneumoniae strain AWD5, isolated from an automobile workshop in India. The de novo assembly resulted in a 4,807,409 bp genome containing 25 rRNA genes, 81 tRNAs, and 4,636 coding sequences (CDS). It carries important genes for polyaromatic hydrocarbon degradation and benzoate degradation. PMID:28153891

  13. Isolation and characterization of Klebsiella pneumoniae unencapsulated mutants

    SciTech Connect

    Benedi, V.J.; Ciurana, B.; Tomas, J.M.

    1989-01-01

    Klebsiella pneumoniae mutants were obtained after UV irradiation and negative selection with anticapsular serum. Unencapsulation, rather than expression of a structurally altered capsule, was found in the mutants. The mutant strains showed no alterations in their outer membrane proteins and lipopolysaccharide, and a great similarity with the wild type in the properties tested (serum resistance, antimicrobial sensitivity, and lipopolysaccharide-specific bacteriophage sensitivity), with the exception of a higher cell surface hydrophobicity and resistance to bacteriophage FC3-9.

  14. Klebsiella pneumonia with pneumatocele formation in a newborn infant

    PubMed Central

    Papageorgiou, Apostolos; Bauer, Charles R.; Fletcher, Barry D.; Stern, Leo

    1973-01-01

    The case of a premature infant with bacteriologically proved klebsiella pneumonia in the first week of life is reported. The clinical course was complicated by the early formation of pneumatoceles and subsequent recurrent pneumothoraces. Treatment consisted of antibiotic therapy and chest tube drainage. Satisfactory clinical recovery, growth and psychomotor development were noted at 1 year of age, despite the persistence of a pneumatocele in the right lung. ImagesFIG. 2FIG. 3FIG. 4 PMID:4586073

  15. Computed tomographic differences of pyogenic liver abscesses caused by Klebsiella pneumoniae and non-Klebsiella pneumoniae.

    PubMed

    Kim, Sung-Bum; Je, Bo-Kyung; Lee, Kee Yeol; Lee, Seung Hwa; Chung, Hwan-Hoon; Cha, Sang Hoon

    2007-01-01

    To investigate the distinctive computed tomographic(CT) features of pyogenic liver abscess (PLA) caused by Klebsiella pneumoniae (KLA). 114 PLA patients (62 men, 52 women, age range 25-89 years, mean age 60.9 years) with positive yield of blood or pus cultures were divided into 2 groups; KLA group (n = 58) and non-KLA group (n = 56). We retrospectively reviewed the CT images. The independent-sample t-test, X2 test, or Fisher's exact test were used with SPSS for windows version 11.0. Gas formation showed no significant difference between two groups (P = 0.284), but air-fluid pattern was found only in KLA group (P = 0.027). The rim on dynamic CT was showed less frequently in KLA group than non-KLA group (P < 0.001), and smooth and regular rim was more common in non-KLA group (P < 0.001). The septation and septal breakage were more common in KLA group (P < 0.001). The turquoise sign was found significantly more in KLA group (P < 0.001). The hairball sign was detected only in KLA group (P < 0.001). The cluster sign was less frequent in KLA group (P < 0.001). KLA had a series of characteristic intra-abscess features; septal breakage, turquoise-like structure, hairball-like content, air-fluid level, and no enhanced rim. When turquoise/hairball sign or air-fluid level is present in abscess, KLA would be a probable diagnosis in the clinical field.

  16. Antimicrobial resistance temporal trend of Klebsiella pneumoniae isolated from blood

    PubMed Central

    Gavriliu, LC; Benea, OE; Benea, S

    2016-01-01

    Background. According to the European Antimicrobial Resistance Surveillance Network, Romania reports an increasing number of resistant Klebsiella pneumoniae strains from invasive infections every year. Material and Method. We analyzed the antimicrobial susceptibility of Klebsiella pneumoniae strains isolated from blood in 2010 and 2015 in “Matei Bals” National Institute of Infectious Diseases, in order to identify any significant changes in the last five years. Results. We identified 18 strains in 2010 and 37 strains in 2015. Although the resistance to aminopenicillin-betalactamase inhibitors association, piperacillin-tazobactam, third generation cephalosporins, fluoroquinolones, gentamicin, amikacin and the combined resistance decreased between these two time frames, this evolution was statistically non-significant. The same was noticed for the increased resistance rates to carbapenems. Conclusions. Antimicrobial resistance of Klebsiella pneumoniae may become a major problem for the public health and the hospital-acquired infections control. Therefore, it needs further monitoring and efforts must be made in order to limit the increase of the resistance. PMID:27928448

  17. Modulation of respiratory dendritic cells during Klebsiella pneumonia infection

    PubMed Central

    2013-01-01

    Background Klebsiella pneumoniae is a leading cause of severe hospital-acquired respiratory tract infections and death but little is known regarding the modulation of respiratory dendritic cell (DC) subsets. Plasmacytoid DC (pDC) are specialized type 1 interferon producing cells and considered to be classical mediators of antiviral immunity. Method By using multiparameter flow cytometry analysis we have analysed the modulation of respiratory DC subsets after intratracheal Klebsiella pneumonia infection. Results Data indicate that pDCs and MoDC were markedly elevated in the post acute pneumonia phase when compared to mock-infected controls. Analysis of draining mediastinal lymph nodes revealed a rapid increase of activated CD103+ DC, CD11b+ DC and MoDC within 48 h post infection. Lung pDC identification during bacterial pneumonia was confirmed by extended phenotyping for 120G8, mPDCA-1 and Siglec-H expression and by demonstration of high Interferon-alpha producing capacity after cell sorting. Cytokine expression analysis of ex vivo-sorted respiratory DC subpopulations from infected animals revealed elevated Interferon-alpha in pDC, elevated IFN-gamma, IL-4 and IL-13 in CD103+ DC and IL-19 and IL-12p35 in CD11b+ DC subsets in comparison to CD11c+ MHC-class IIlow cells indicating distinct functional roles. Antigen-specific naive CD4+ T cell stimulatory capacity of purified respiratory DC subsets was analysed in a model system with purified ovalbumin T cell receptor transgenic naive CD4+ responder T cells and respiratory DC subsets, pulsed with ovalbumin and matured with Klebsiella pneumoniae lysate. CD103+ DC and CD11b+ DC subsets represented the most potent naive CD4+ T helper cell activators. Conclusion These results provide novel insight into the activation of respiratory DC subsets during Klebsiella pneumonia infection. The detection of increased respiratory pDC numbers in bacterial pneumonia may indicate possible novel pDC functions with respect to lung repair

  18. Modulation of respiratory dendritic cells during Klebsiella pneumonia infection.

    PubMed

    Hackstein, Holger; Kranz, Sabine; Lippitsch, Anne; Wachtendorf, Andreas; Kershaw, Olivia; Gruber, Achim D; Michel, Gabriela; Lohmeyer, Jürgen; Bein, Gregor; Baal, Nelli; Herold, Susanne

    2013-09-17

    Klebsiella pneumoniae is a leading cause of severe hospital-acquired respiratory tract infections and death but little is known regarding the modulation of respiratory dendritic cell (DC) subsets. Plasmacytoid DC (pDC) are specialized type 1 interferon producing cells and considered to be classical mediators of antiviral immunity. By using multiparameter flow cytometry analysis we have analysed the modulation of respiratory DC subsets after intratracheal Klebsiella pneumonia infection. Data indicate that pDCs and MoDC were markedly elevated in the post acute pneumonia phase when compared to mock-infected controls. Analysis of draining mediastinal lymph nodes revealed a rapid increase of activated CD103+ DC, CD11b+ DC and MoDC within 48 h post infection. Lung pDC identification during bacterial pneumonia was confirmed by extended phenotyping for 120G8, mPDCA-1 and Siglec-H expression and by demonstration of high Interferon-alpha producing capacity after cell sorting. Cytokine expression analysis of ex vivo-sorted respiratory DC subpopulations from infected animals revealed elevated Interferon-alpha in pDC, elevated IFN-gamma, IL-4 and IL-13 in CD103+ DC and IL-19 and IL-12p35 in CD11b+ DC subsets in comparison to CD11c+ MHC-class IIlow cells indicating distinct functional roles. Antigen-specific naive CD4+ T cell stimulatory capacity of purified respiratory DC subsets was analysed in a model system with purified ovalbumin T cell receptor transgenic naive CD4+ responder T cells and respiratory DC subsets, pulsed with ovalbumin and matured with Klebsiella pneumoniae lysate. CD103+ DC and CD11b+ DC subsets represented the most potent naive CD4+ T helper cell activators. These results provide novel insight into the activation of respiratory DC subsets during Klebsiella pneumonia infection. The detection of increased respiratory pDC numbers in bacterial pneumonia may indicate possible novel pDC functions with respect to lung repair and regeneration.

  19. Metabolism of benzonitrile and butyronitrile by Klebsiella pneumoniae

    SciTech Connect

    Nawaz, M.S.; Heinze, T.M.; Cerniglia, C.E. )

    1992-01-01

    A strain of Klebsiella pneumoniae that used aliphatic nitriles as the sole source of nitrogen was adapted to benzonitrile as the sole source of carbon and nitrogen. Gas chromatographic and mass spectral analyses of culture filtrates indicated that K. pneumoniae metabolized 8.4 mM benzonitrile to 4.0 mM benzoic acid and 2.7 mM ammonia. In addition, butyronitrile was metabolized to butyramide and ammonia. The isolate also degraded mixtures of benzonitrile and aliphatic nitriles. Cell extracts contained nitrile hydratase and amidase activities. The enzyme activities were higher with butyronitrile and butyramide than with benzonitrile and benzamide, and amidase activities were twofold higher than nitrile hydratase activities. K. pneumoniae appears promising for the bioremediation of sites contaminated with aliphatic and aromatic nitriles.

  20. Metabolism of benzonitrile and butyronitrile by Klebsiella pneumoniae.

    PubMed Central

    Nawaz, M S; Heinze, T M; Cerniglia, C E

    1992-01-01

    A strain of Klebsiella pneumoniae that used aliphatic nitriles as the sole source of nitrogen was adapted to benzonitrile as the sole source of carbon and nitrogen. Gas chromatographic and mass spectral analyses of culture filtrates indicated that K. pneumoniae metabolized 8.4 mM benzonitrile to 4.0 mM benzoic acid and 2.7 mM ammonia. In addition, butyronitrile was metabolized to butyramide and ammonia. The isolate also degraded mixtures of benzonitrile and aliphatic nitriles. Cell extracts contained nitrile hydratase and amidase activities. The enzyme activities were higher with butyronitrile and butyramide than with benzonitrile and benzamide, and amidase activities were twofold higher than nitrile hydratase activities. K. pneumoniae appears promising for the bioremediation of sites contaminated with aliphatic and aromatic nitriles. PMID:1539979

  1. Klebsiella pneumoniae: Going on the Offense with a Strong Defense

    PubMed Central

    Paczosa, Michelle K.

    2016-01-01

    SUMMARY Klebsiella pneumoniae causes a wide range of infections, including pneumonias, urinary tract infections, bacteremias, and liver abscesses. Historically, K. pneumoniae has caused serious infection primarily in immunocompromised individuals, but the recent emergence and spread of hypervirulent strains have broadened the number of people susceptible to infections to include those who are healthy and immunosufficient. Furthermore, K. pneumoniae strains have become increasingly resistant to antibiotics, rendering infection by these strains very challenging to treat. The emergence of hypervirulent and antibiotic-resistant strains has driven a number of recent studies. Work has described the worldwide spread of one drug-resistant strain and a host defense axis, interleukin-17 (IL-17), that is important for controlling infection. Four factors, capsule, lipopolysaccharide, fimbriae, and siderophores, have been well studied and are important for virulence in at least one infection model. Several other factors have been less well characterized but are also important in at least one infection model. However, there is a significant amount of heterogeneity in K. pneumoniae strains, and not every factor plays the same critical role in all virulent Klebsiella strains. Recent studies have identified additional K. pneumoniae virulence factors and led to more insights about factors important for the growth of this pathogen at a variety of tissue sites. Many of these genes encode proteins that function in metabolism and the regulation of transcription. However, much work is left to be done in characterizing these newly discovered factors, understanding how infections differ between healthy and immunocompromised patients, and identifying attractive bacterial or host targets for treating these infections. PMID:27307579

  2. Killing of Klebsiella pneumoniae by human alveolar macrophages.

    PubMed

    Hickman-Davis, Judy M; O'Reilly, Philip; Davis, Ian C; Peti-Peterdi, Janos; Davis, Glenda; Young, K Randall; Devlin, Robert B; Matalon, Sadis

    2002-05-01

    We investigated putative mechanisms by which human surfactant protein A (SP-A) effects killing of Klebsiella pneumoniae by human alveolar macrophages (AMs) isolated from bronchoalveolar lavagates of patients with transplanted lungs. Coincubation of AMs with human SP-A (25 microg/ml) and Klebsiella resulted in a 68% decrease in total colony forming units by 120 min compared with AMs infected with Klebsiella in the absence of SP-A, and this SP-A-mediated effect was abolished by preincubation with N(G)-monomethyl-L-arginine. Incubation of transplant AMs with SP-A increased intracellular Ca(2+) concentration ([Ca(2+)](i)) by 70% and nitrite and nitrate (NO(x)) production by 45% (from 0.24 +/- 0.02 to 1.3 +/- 0.21 nmol small middle dot 10(6) AMs(-1).h(-1)). Preincubation with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester inhibited the increase in [Ca(2+)](i) and abrogated the SP-A-mediated Klebsiella phagocytosis and killing. In contrast, incubation of AMs from normal volunteers with SP-A decreased both [Ca(2+)](i) and NO(x) production and did not result in killing of Klebsiella. Significant killing of Klebsiella was also seen in a cell-free system by sustained production of peroxynitrite (>1 microM/min) at pH 5 but not at pH 7.4. These findings indicate that SP-A mediates pathogen killing by AMs from transplant lungs by stimulating phagocytosis and production of reactive oxygen-nitrogen intermediates.

  3. Deciphering tissue-induced Klebsiella pneumoniae lipid A structure.

    PubMed

    Llobet, Enrique; Martínez-Moliner, Verónica; Moranta, David; Dahlström, Käthe M; Regueiro, Verónica; Tomás, Anna; Cano, Victoria; Pérez-Gutiérrez, Camino; Frank, Christian G; Fernández-Carrasco, Helena; Insua, José Luis; Salminen, Tiina A; Garmendia, Junkal; Bengoechea, José A

    2015-11-17

    The outcome of an infection depends on host recognition of the pathogen, hence leading to the activation of signaling pathways controlling defense responses. A long-held belief is that the modification of the lipid A moiety of the lipopolysaccharide could help Gram-negative pathogens to evade innate immunity. However, direct evidence that this happens in vivo is lacking. Here we report the lipid A expressed in the tissues of infected mice by the human pathogen Klebsiella pneumoniae. Our findings demonstrate that Klebsiella remodels its lipid A in a tissue-dependent manner. Lipid A species found in the lungs are consistent with a 2-hydroxyacyl-modified lipid A dependent on the PhoPQ-regulated oxygenase LpxO. The in vivo lipid A pattern is lost in minimally passaged bacteria isolated from the tissues. LpxO-dependent modification reduces the activation of inflammatory responses and mediates resistance to antimicrobial peptides. An lpxO mutant is attenuated in vivo thereby highlighting the importance of this lipid A modification in Klebsiella infection biology. Colistin, one of the last options to treat multidrug-resistant Klebsiella infections, triggers the in vivo lipid A pattern. Moreover, colistin-resistant isolates already express the in vivo lipid A pattern. In these isolates, LpxO-dependent lipid A modification mediates resistance to colistin. Deciphering the lipid A expressed in vivo opens the possibility of designing novel therapeutics targeting the enzymes responsible for the in vivo lipid A pattern.

  4. Deciphering tissue-induced Klebsiella pneumoniae lipid A structure

    PubMed Central

    Llobet, Enrique; Martínez-Moliner, Verónica; Moranta, David; Dahlström, Käthe M.; Regueiro, Verónica; Tomás, Anna; Cano, Victoria; Pérez-Gutiérrez, Camino; Frank, Christian G.; Fernández-Carrasco, Helena; Insua, José Luis; Salminen, Tiina A.; Garmendia, Junkal; Bengoechea, José A.

    2015-01-01

    The outcome of an infection depends on host recognition of the pathogen, hence leading to the activation of signaling pathways controlling defense responses. A long-held belief is that the modification of the lipid A moiety of the lipopolysaccharide could help Gram-negative pathogens to evade innate immunity. However, direct evidence that this happens in vivo is lacking. Here we report the lipid A expressed in the tissues of infected mice by the human pathogen Klebsiella pneumoniae. Our findings demonstrate that Klebsiella remodels its lipid A in a tissue-dependent manner. Lipid A species found in the lungs are consistent with a 2-hydroxyacyl-modified lipid A dependent on the PhoPQ-regulated oxygenase LpxO. The in vivo lipid A pattern is lost in minimally passaged bacteria isolated from the tissues. LpxO-dependent modification reduces the activation of inflammatory responses and mediates resistance to antimicrobial peptides. An lpxO mutant is attenuated in vivo thereby highlighting the importance of this lipid A modification in Klebsiella infection biology. Colistin, one of the last options to treat multidrug-resistant Klebsiella infections, triggers the in vivo lipid A pattern. Moreover, colistin-resistant isolates already express the in vivo lipid A pattern. In these isolates, LpxO-dependent lipid A modification mediates resistance to colistin. Deciphering the lipid A expressed in vivo opens the possibility of designing novel therapeutics targeting the enzymes responsible for the in vivo lipid A pattern. PMID:26578797

  5. Lipopolysaccharide-specific bacteriophage for Klebsiella pneumoniae C3.

    PubMed Central

    Tomás, J M; Jofre, J T

    1985-01-01

    Bacteriophage FC3-1 is one of several specific bacteriophages of Klebsiella pneumoniae C3 isolated in our laboratory. Unlike receptors for other Klebsiella phages, the bacteriophage FC3-1 receptor was shown to be lipopolysaccharide, specifically the polysaccharide fraction (O-antigen and core region). We concluded that capsular polysaccharide, outer membrane proteins, and lipid A were not involved in phage binding. Mutants resistant to this phage were isolated and were found to be devoid of lipopolysaccharide O-antigen by several criteria but to contain capsular material serologically identical to that of the wild type. The polysaccharide fraction was concluded to be the primary phage receptor, indicating that it is available to the phage. Images PMID:3888963

  6. Isolation and characterisation of lytic bacteriophages of Klebsiella pneumoniae and Klebsiella oxytoca.

    PubMed

    Karumidze, Natia; Kusradze, Ia; Rigvava, Sophio; Goderdzishvili, Marine; Rajakumar, Kumar; Alavidze, Zemphira

    2013-03-01

    Klebsiella bacteria have emerged as an increasingly important cause of community-acquired nosocomial infections. Extensive use of broad-spectrum antibiotics in hospitalised patients has led to both increased carriage of Klebsiella and the development of multidrug-resistant strains that frequently produce extended-spectrum β-lactamases and/or other defences against antibiotics. Many of these strains are highly virulent and exhibit a strong propensity to spread. In this study, six lytic Klebsiella bacteriophages were isolated from sewage-contaminated river water in Georgia and characterised as phage therapy candidates. Two of the phages were investigated in greater detail. Biological properties, including phage morphology, nucleic acid composition, host range, growth phenotype, and thermal and pH stability were studied for all six phages. Limited sample sequencing was performed to define the phylogeny of the K. pneumoniae- and K. oxytoca-specific bacteriophages vB_Klp_5 and vB_Klox_2, respectively. Both of the latter phages had large burst sizes, efficient rates of adsorption and were stable under different adverse conditions. Phages reported in this study are double-stranded DNA bacterial viruses belonging to the families Podoviridae and Siphoviridae. One or more of the six phages was capable of efficiently lysing ~63 % of Klebsiella strains comprising a collection of 123 clinical isolates from Georgia and the United Kingdom. These phages exhibit a number of properties indicative of potential utility in phage therapy cocktails.

  7. Klebsiella pneumonia liver abscess syndrome: Case presentation to a college student health clinic.

    PubMed

    Woll, Christopher; Spotts, P Hunter

    2016-01-01

    The authors describe a case of Klebsiella pneumoniae liver abscess (KPLA) in a student presenting to a university student health center. The authors also provide a review of KPLA and invasive Klebsiella pneumoniae liver abscess syndrome (IKPLAS), including epidemiology, common clinical manifestations, standard diagnostic work-up, management options, and potential complications.

  8. Klebsiella pneumoniae carrying bla NDM-1 gene in orthopedic practice.

    PubMed

    Gupta, Varsha; Bansal, Neha; Gupta, Ravi; Chander, Jagdish

    2014-09-01

    Emergence and spread of carbapenemases in Enterobacteriaceae is a cause of concern worldwide, the latest threat being New Delhi metallo-β-lactamase (NDM-1). This report is of an orthopedic case with fracture femur managed with internal fixation and bone grafting, who subsequently developed secondary infection with Klebsiella pneumoniae harboring bla NDM-1 gene. Minimum inhibitory concentration (MIC) of imipenem was ≥8 μg/ml by E-test, suggestive of carbapenemase production. Phenotypic and further genotypic detection confirmed the presence of bla NDM-1 gene. The isolate remained susceptible only to tigecycline, colistin, and polymyxin B.

  9. Acquisitive evolution of ribitol dehydrogenase in Klebsiella pneumoniae.

    PubMed Central

    Thompson, L W; Krawiec, S

    1983-01-01

    Selection in continuous culture of Klebsiella pneumoniae mutants that have gained the ability to utilize xylitol while also retaining regulatory control over ribitol utilization was achieved with a dual-substrate regime. Initial steady-state cultures of wild-type organisms were maintained with 0.005% (0.329 mM) ribitol. Mutants of various types proliferated when the composition of the limiting medium was changed to 0.005% ribitol plus 0.250% (16.43 mM) xylitol. PMID:6341353

  10. Clinical epidemiology of the global expansion of Klebsiella pneumoniae carbapenemases

    PubMed Central

    Munoz-Price, L Silvia; Poirel, Laurent; Bonomo, Robert A; Schwaber, Mitchell J; Daikos, George L; Cormican, Martin; Cornaglia, Giuseppe; Garau, Javier; Gniadkowski, Marek; Hayden, Mary K; Kumarasamy, Karthikeyan; Livermore, David M; Maya, Juan J; Nordmann, Patrice; Patel, Jean B; Paterson, David L; Pitout, Johann; Villegas, Maria Virginia; Wang, Hui; Woodford, Neil; Quinn, John P

    2015-01-01

    Klebsiella pneumoniae carbapenemases (KPCs) were originally identified in the USA in 1996. Since then, these versatile β-lactamases have spread internationally among Gram-negative bacteria, especially K pneumoniae, although their precise epidemiology is diverse across countries and regions. The mortality described among patients infected with organisms positive for KPC is high, perhaps as a result of the limited antibiotic options remaining (often colistin, tigecycline, or aminoglycosides). Triple drug combinations using colistin, tigecycline, and imipenem have recently been associated with improved survival among patients with bacteraemia. In this Review, we summarise the epidemiology of KPCs across continents, and discuss issues around detection, present antibiotic options and those in development, treatment outcome and mortality, and infection control. In view of the limitations of present treatments and the paucity of new drugs in the pipeline, infection control must be our primary defence for now. PMID:23969216

  11. Granzymes A and B Regulate the Local Inflammatory Response during Klebsiella pneumoniae Pneumonia.

    PubMed

    García-Laorden, M Isabel; Stroo, Ingrid; Blok, Dana C; Florquin, Sandrine; Medema, Jan Paul; de Vos, Alex F; van der Poll, Tom

    2016-01-01

    Klebsiella pneumoniae is a common cause of hospital-acquired pneumonia. Granzymes (gzms), mainly found in cytotoxic lymphocytes, have been implicated as mediators of infection and inflammation. We here sought to investigate the role of gzmA and gzmB in the host response to K. pneumoniae-induced airway infection and sepsis. For this purpose, pneumonia was induced in wild-type (WT) and gzmA-deficient (gzmA-/-), gzmB-/- and gzmAxB-/- mice by intranasal infection with K. pneumoniae. In WT mice, gzmA and gzmB were mainly expressed by natural killer cells. Pneumonia was associated with reduced intracellular gzmA and increased intracellular gzmB levels. Gzm deficiency had little impact on antibacterial defence: gzmA-/- and gzmAxB-/- mice transiently showed modestly higher bacterial loads in the lungs but not in distant organs. GzmB-/- and, to a larger extent, gzmAxB-/- mice displayed transiently increased lung inflammation, reflected in the semi-quantitative histology scores and levels of pro-inflammatory cytokines and chemokines. Most differences between gzm-deficient and WT mice had disappeared during late-stage pneumonia. Gzm deficiency did not impact on distant organ injury or survival. These results suggest that gzmA and gzmB partly regulate local inflammation during early pneumonia but eventually play an insignificant role during pneumosepsis by the common human pathogen K. pneumoniae.

  12. CRYSTAL STRUCTURE ANALYSIS OF A PUTATIVE OXIDOREDUCTASE FROM KLEBSIELLA PNEUMONIAE

    SciTech Connect

    Baig, M.; Brown, A.; Eswaramoorthy, S.; Swaminathan, S.

    2009-01-01

    Klebsiella pneumoniae, a gram-negative enteric bacterium, is found in nosocomial infections which are acquired during hospital stays for about 10% of hospital patients in the United States. The crystal structure of a putative oxidoreductase from K. pneumoniae has been determined. The structural information of this K. pneumoniae protein was used to understand its function. Crystals of the putative oxidoreductase enzyme were obtained by the sitting drop vapor diffusion method using Polyethylene glycol (PEG) 3350, Bis-Tris buffer, pH 5.5 as precipitant. These crystals were used to collect X-ray data at beam line X12C of the National Synchrotron Light Source (NSLS) at Brookhaven National Laboratory (BNL). The crystal structure was determined using the SHELX program and refi ned with CNS 1.1. This protein, which is involved in the catalysis of an oxidation-reduction (redox) reaction, has an alpha/beta structure. It utilizes nicotinamide adenine dinucleotide phosphate (NADP) or nicotine adenine dinucleotide (NAD) to perform its function. This structure could be used to determine the active and co-factor binding sites of the protein, information that could help pharmaceutical companies in drug design and in determining the protein’s relationship to disease treatment such as that for pneumonia and other related pathologies.

  13. Molecular Epidemiology of Colonizing and Infecting Isolates of Klebsiella pneumoniae

    PubMed Central

    Martin, Rebekah M.; Cao, Jie; Brisse, Sylvain; Passet, Virginie; Wu, Weisheng; Zhao, Lili; Malani, Preeti N.; Rao, Krishna

    2016-01-01

    ABSTRACT Klebsiella pneumoniae is among the most common causes of hospital-acquired infections and has emerged as an urgent threat to public health due to carbapenem antimicrobial resistance. K. pneumoniae commonly colonizes hospitalized patients and causes extraintestinal infections such as urinary tract infection, bloodstream infection (septicemia), and pneumonia. If colonization is an intermediate step before infection, then detection and characterization of colonizing isolates could enable strategies to prevent or empirically treat K. pneumoniae infections in hospitalized patients. However, the strength of the association between colonization and infection is unclear. To test the hypothesis that hospitalized patients become infected with their colonizing strain, 1,765 patients were screened for rectal colonization with K. pneumoniae, and extraintestinal isolates from these same patients were collected over a 3-month period in a cohort study design. The overall colonization prevalence was 23.0%. After adjustment for other patient factors, colonization was significantly associated with subsequent infection: 21 of 406 (5.2%) colonized patients later had extraintestinal infection, compared to 18 of 1,359 (1.3%) noncolonized patients (adjusted odds ratio [OR], 4.01; 95% confidence interval, 2.08 to 7.73; P < 0.001). Despite a high diversity of colonizing isolates, 7/7 respiratory, 4/4 urinary, and 2/5 bloodstream isolates from colonized patients matched the patient corresponding rectal swab isolates, based on wzi capsular typing, multilocus sequence typing (MLST), and whole-genome sequence analysis. These results suggest that K. pneumoniae colonization is directly associated with progression to extraintestinal infection. IMPORTANCE K. pneumoniae commonly infects hospitalized patients, and these infections are increasingly resistant to carbapenems, the antibiotics of last resort for life-threatening bacterial infections. To prevent and treat these infections, we

  14. Molecular Epidemiology of Colonizing and Infecting Isolates of Klebsiella pneumoniae.

    PubMed

    Martin, Rebekah M; Cao, Jie; Brisse, Sylvain; Passet, Virginie; Wu, Weisheng; Zhao, Lili; Malani, Preeti N; Rao, Krishna; Bachman, Michael A

    2016-01-01

    Klebsiella pneumoniae is among the most common causes of hospital-acquired infections and has emerged as an urgent threat to public health due to carbapenem antimicrobial resistance. K. pneumoniae commonly colonizes hospitalized patients and causes extraintestinal infections such as urinary tract infection, bloodstream infection (septicemia), and pneumonia. If colonization is an intermediate step before infection, then detection and characterization of colonizing isolates could enable strategies to prevent or empirically treat K. pneumoniae infections in hospitalized patients. However, the strength of the association between colonization and infection is unclear. To test the hypothesis that hospitalized patients become infected with their colonizing strain, 1,765 patients were screened for rectal colonization with K. pneumoniae, and extraintestinal isolates from these same patients were collected over a 3-month period in a cohort study design. The overall colonization prevalence was 23.0%. After adjustment for other patient factors, colonization was significantly associated with subsequent infection: 21 of 406 (5.2%) colonized patients later had extraintestinal infection, compared to 18 of 1,359 (1.3%) noncolonized patients (adjusted odds ratio [OR], 4.01; 95% confidence interval, 2.08 to 7.73; P < 0.001). Despite a high diversity of colonizing isolates, 7/7 respiratory, 4/4 urinary, and 2/5 bloodstream isolates from colonized patients matched the patient corresponding rectal swab isolates, based on wzi capsular typing, multilocus sequence typing (MLST), and whole-genome sequence analysis. These results suggest that K. pneumoniae colonization is directly associated with progression to extraintestinal infection. IMPORTANCE K. pneumoniae commonly infects hospitalized patients, and these infections are increasingly resistant to carbapenems, the antibiotics of last resort for life-threatening bacterial infections. To prevent and treat these infections, we must

  15. Preliminary investigation of a mice model of Klebsiella pneumoniae subsp. ozaenae induced pneumonia.

    PubMed

    Renois, Fanny; Jacques, Jérôme; Guillard, Thomas; Moret, Hélène; Pluot, Michel; Andreoletti, Laurent; de Champs, Christophe

    2011-11-01

    In the present study, we comparatively assessed the pathophysiological mechanisms developed during lung infection of BALB/C female mice infected by an original wild type Klebsiella pneumoniae subsp. ozaenae strain (CH137) or by a referent subspecies K. pneumoniae. subsp. pneumoniae strain (ATCC10031). The mice infected with 2.10⁶ CFU K. p. subsp. pneumoniae (n = 10) showed transient signs of infection and all of them recovered. All of those infected with 1.10⁶ CFU K. p. subsp. ozaenae (n = 10) developed pneumonia within 24 h and died between 48 and 72 h. Few macrophages, numerous polymorphonuclear cells and lymphocytes were observed in their lungs in opposite to K. p. subsp. pneumoniae. In bronchoalveolar lavage, a significant increase in MIP-2, IL-6, KC and MCP-1 levels was only observed in K. p. subsp. ozaenae infected mice whereas high levels of TNF-α were evidenced with the two subspecies. Our findings indicated a lethal effect of a wild type K. p. subsp. ozaenae strain by acute pneumonia reflecting an insufficient alveolar macrophage response. This model might be of a major interest to comparatively explore the pathogenicity of K. p. subsp ozaenae strains and to further explore the physiopathological mechanisms of gram-negative bacteria induced human pneumonia. Copyright © 2011 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  16. Risk Factors and Clinical Impact of Klebsiella pneumoniae Carbapenemase–Producing K. pneumoniae

    PubMed Central

    Gasink, Leanne B.; Edelstein, Paul H.; Lautenbach, Ebbing; Synnestvedt, Marie; Fishman, Neil O.

    2010-01-01

    BACKGROUND Klebsiella pneumoniae carbapenemase (KPC)–producing K. pneumoniae is an emerging pathogen with serious clinical and infection control implications. To our knowledge, no study has specifically examined risk factors for KPC-producing K. pneumoniae or its impact on mortality. METHODS To identify risk factors for infection or colonization with KPC-producing K. pneumoniae, a case-control study was performed. Case patients with KPC-producing K. pneumoniae were compared with control subjects with carbapenem-susceptible K. pneumoniae. A cohort study evaluated the association between KPC-producing K. pneumoniae and in-hospital mortality. RESULTS Fifty-six case patients and 863 control subjects were identified. In multivariable analysis, independent risk factors for KPC-producing K. pneumoniae were (1) severe illness (adjusted odds ratio [AOR], 4.31; 95% confidence interval [CI], 2.25–8.25), (2) prior fluoroquinolone use (AOR, 3.39; 95% CI, 1.50, 7.66), and (3) prior extended-spectrum cephalosporin use (AOR, 2.55; 95% CI, 1.18, 5.52). Compared with samples from other anatomic locations, K. pneumoniae isolates from blood samples were less likely to harbor KPC (AOR, 0.33; 95% CI, 0.12, 0.86). KPC-producing K. pneumoniae was independently associated with in-hospital mortality (AOR, 3.60; 95% CI, 1.87–6.91). CONCLUSIONS KPC-producing K. pneumoniae is an emerging pathogen associated with significant mortality. Our findings highlight the urgent need to develop strategies for prevention and infection control. Limiting use of certain antimicrobials, specifically fluoroquinolones and cephalosporins, use may be effective strategies. PMID:19860564

  17. Biofilm formation and antibiotic resistance in Klebsiella pneumoniae urinary strains.

    PubMed

    Vuotto, C; Longo, F; Pascolini, C; Donelli, G; Balice, M P; Libori, M F; Tiracchia, V; Salvia, A; Varaldo, P E

    2017-10-01

    Multidrug-resistant Klebsiella pneumoniae has become a relevant healthcare-associated pathogen. Capsule, type 1 and 3 fimbriae (mrkA gene), type 2 quorum-sensing system (luxS), synthesis of D-galactan I (wbbM), LPS transport (wzm) and poly-beta-1,6-N-acetyl-D-glucosamine (pgaA) seem involved in K. pneumoniae biofilm. Nonenzymatic antibiotic resistance is related to nonexpression or mutation of porins (OmpK35 and OmpK36), and efflux pump (acrB) overexpression. The aim of this study was to analyse some virulence factors of K. pneumoniae isolates, and to evaluate possible correlations between their antibiotic resistance profile and ability to form biofilm. Quantitative biofilm production assay, congo red agar test and string test were performed on 120 isolates clustered in 56 extensively drug-resistant (XDR), 40 MDR and 24 susceptible (S) strains. Nine representative strains were analysed by real-time RT-PCR for the expression of antibiotic resistance (OmpK35, OmpK36, acrB) and biofilm production genes (mrkA, luxS, pga, wbbM, wzm) during planktonic and sessile growth. XDR isolates showed a higher ability to form biofilm (91·07%) and to produce polysaccharides (78·57%) when compared to MDR and S strains. In biofilm-growing XDR strains, seven of eight genes were upregulated, with the only exception of OmpK36. XDR strains exhibited phenotypic and genotypic features supporting a significant growth as biofilm. This study produces new findings that highlight a positive correlation between antibiotic resistance profile and biofilm-forming ability in XDR K. pneumoniae strains. These new evidences might contribute to the progress in selection of therapeutic treatments of infections caused by K. pneumoniae resistant also to the 'last line of defence' antibiotics, that is, carbapenems. © 2017 The Society for Applied Microbiology.

  18. Biological Treatment of Cyanide by Using
Klebsiella pneumoniae Species

    PubMed Central

    Bilkay, Isil Seyis

    2016-01-01

    Summary In this study, optimization conditions for cyanide biodegradation by Klebsiella pneumoniae strain were determined to be 25 °C, pH=7 and 150 rpm at the concentration of 0.5 mM potassium cyanide in the medium. Additionally, it was found that K. pneumoniae strain is not only able to degrade potassium cyanide, but also to degrade potassium hexacyanoferrate(II) trihydrate and sodium ferrocyanide decahydrate with the efficiencies of 85 and 87.5%, respectively. Furthermore, this strain degraded potassium cyanide in the presence of different ions such as magnesium, nickel, cobalt, iron, chromium, arsenic and zinc, in variable concentrations (0.1, 0.25 and 0.5 mM) and as a result the amount of the bacteria in the biodegradation media decreased with the increase of ion concentration. Lastly, it was also observed that sterile crude extract of K. pneumoniae strain degraded potassium cyanide on the fifth day of incubation. Based on these results, it is concluded that both culture and sterile crude extract of K. pnemoniae will be used in cyanide removal from different wastes. PMID:28115902

  19. Purification and properties of dihydroxyacetone kinase from Klebsiella pneumoniae.

    PubMed Central

    Johnson, E A; Burke, S K; Forage, R G; Lin, E C

    1984-01-01

    Dihydroxyacetone (DHA) kinase of Klebsiella pneumoniae, a gene product of the dha regulon responsible for fermentative dissimilation of glycerol and DHA, was purified 120-fold to a final specific activity of 10 mumol X min-1 X mg of protein-1 at 30 degrees C. The enzyme, a dimer of a 53,000 +/- 5,000-dalton polypeptide, is highly specific for DHA (Km, ca.4 microM). Glycerol is not a substrate at 1 mM and is not an inhibitor even at 100 mM. The enzyme is not inhibited by 5 mM fructose-1,6-diphosphate. Ca2+ gives a higher enzyme activity than Mg2+ as a cationic cofactor. Escherichia coli glycerol kinase acts on both glycerol and DHA and is allosterically inhibited by fructose-1,6-diphosphate. Antibodies raised against E. coli glycerol kinase cross-reacted with K. pneumoniae glycerol kinase but not with K. pneumoniae DHA kinase. Images PMID:6090436

  20. Hypervirulent (hypermucoviscous) Klebsiella pneumoniae: a new and dangerous breed.

    PubMed

    Shon, Alyssa S; Bajwa, Rajinder P S; Russo, Thomas A

    2013-02-15

    A new hypervirulent (hypermucoviscous) variant of Klebsiella pneumoniae has emerged. First described in the Asian Pacific Rim, it now increasingly recognized in Western countries. Defining clinical features are the ability to cause serious, life-threatening community-acquired infection in younger healthy hosts, including liver abscess, pneumonia, meningitis and endophthalmitis and the ability to metastatically spread, an unusual feature for enteric Gram-negative bacilli in the non-immunocompromised. Despite infecting a healthier population, significant morbidity and mortality occurs. Although epidemiologic features are still being defined, colonization, particularly intestinal colonization, appears to be a critical step leading to infection. However the route of entry remains unclear. The majority of cases described to date are in Asians, raising the issue of a genetic predisposition vs. geospecific strain acquisition. The traits that enhance its virulence when compared with "classical" K. pneumoniae are the ability to more efficiently acquire iron and perhaps an increase in capsule production, which confers the hypermucoviscous phenotype. An objective diagnostic test suitable for routine use in the clinical microbiology laboratory is needed. If/when these strains become increasingly resistant to antimicrobials, we will be faced with a frightening clinical scenario.

  1. Epidemiology of Carbapenem Resistant Klebsiella pneumoniae Infections in Mediterranean Countries

    PubMed Central

    Girmenia, Corrado; Serrao, Alessandra; Canichella, Martina

    2016-01-01

    Infections by Carbapenem-Resistant Enterobacteriaceae (CRE), in particular, carbapenem-resistant Klebsiella pneumoniae (CRKp), are a significant public health challenge worldwide. Resistance to carbapenems in enterobacteriaceae is linked to different mechanisms, including the production of the various types of enzymes like KPC, VIM, IMP, NDM, and OXA-48. Despite several attempts to control the spread of these infections at the local and national level, the epidemiological situation for CRKp had worsened in the last years in the Mediterranean area. The rate and types of CRKp isolates greatly differ in the various Mediterranean countries. KPC-producing K. pneumoniae is diffused particularly in the European countries bordering the Mediterranean Sea and is endemic in Greece and Italy. On the contrary, OXA-48-producing K. pneumoniae is endemic in Turkey and Malta and diffused at inter-regional level particularly in some North African and Middle East countries. The spread of these multiresistant pathogens in the world and the Mediterranean countries has been related to various epidemiological factors including the international transfer of patients coming from endemic areas. PMID:27441063

  2. Screening haematology patients for carbapenem-resistant Klebsiella pneumoniae

    PubMed Central

    Kilgour, Elizabeth; Dunn, Caroline; Thomas, Linda; Fox, Richard; Mitchell, Lindsay; Paterson, Pamela

    2013-01-01

    Following a cluster of haematology patients with carbapenem-resistant Klebsiella pneumoniae (CRKP) septicaemia, we initiated screening for rectal carriage of CRKP and multidrug-resistant K. pneumoniae (MDRKP) in this patient group. Haematology inpatients submit a rectal swab once weekly. When plated onto chromogenic Brilliance™ UTI Agar (Oxoid), and incubated overnight with a 10 µg ertapenem disc (Oxoid), K. pneumoniae is identified and semi-automated antibiotic susceptibility testing is performed using the Vitek 2 analyser (Biomerieux). When no zone of inhibition occurs, immediate intervention through patient isolation and enhanced environmental cleaning can be instigated to control further spread while empirical antibiotic prescribing is adapted to take account of identified resistances. Over 2 years, six patients with CRKP and 20 patients with MDRKP were identified. These isolates were resistant to first-line empirical treatment choices for neutropenic sepsis and presented a clinical risk of treatment failure for sepsis post cytotoxic chemotherapy. We describe how this rectal screening methodology was developed and how the results influenced appropriate antibiotic prescribing, patient placement in single rooms and the cleaning of the ward environment to prevent person-to-person transmission of MDRKP and CRKP.

  3. Necrotizing fasciitis caused by hypermucoviscous Klebsiella pneumoniae in a Filipino female in North America.

    PubMed

    Ng, Daniel; Frazee, Brad

    2015-01-01

    Necrotizing fasciitis caused by Klebsiella pneumoniae has been described in Southeast Asia, but has only recently begun to emerge in North America. The hypermucoviscous strain of K. pneumoniae is a particularly virulent strain known to cause devastatingly invasive infections, including necrotizing fasciitis. Here we present the first known case of necrotizing fasciitis caused by hypermucoviscous K. pneumoniae in North America.

  4. Complete genome sequence of a Klebsiella pneumoniae strain isolated from a known cotton insect boll vector

    USDA-ARS?s Scientific Manuscript database

    Klebsiella pneumoniae (associated with bacterial pneumonia) was previously isolated from Nezara viridula, a significant vector of cotton boll-rot pathogens. We provide the first annotated genome sequence of the cotton opportunistic strain K. pneumoniae 5-1. This data provides guidance to study the...

  5. Therapeutic potential of bacteriophage in treating Klebsiella pneumoniae B5055-mediated lobar pneumonia in mice.

    PubMed

    Chhibber, Sanjay; Kaur, Sandeep; Kumari, Seema

    2008-12-01

    Klebsiella pneumoniae causes infections in humans especially in immunocompromised patients. About 80 % of nosocomial infections caused by K. pneumoniae are due to multidrug-resistant strains. The emergence of antibiotic-resistant bacterial strains necessitates the exploration of alternative antibacterial therapies, which led our group to study the ability of bacterial viruses (known as bacteriophages or simply phages) to treat mice challenged with K. pneumoniae. Phage SS specific for K. pneumoniae B5055 was isolated and characterized, and its potential as a therapeutic agent was evaluated in an experimental model of K. pneumoniae-mediated lobar pneumonia in mice. Mice were challenged by intranasal (i.n.) inoculation with bacteria (10(8) c.f.u. ml(-1)). A single intraperitoneal injection of 10(10) p.f.u. ml(-1) phage administered immediately after i.n. challenge was sufficient to rescue 100 % of animals from K. pneumoniae-mediated respiratory infections. Administration of the phage preparation 3 h prior to i.n. bacterial challenge provided significant protection in infected mice, while even 6 h delay of phage administration after the induction of infection rendered the phage treatment ineffective. The results of this study therefore suggest that the timing of starting the phage therapy after initiation of infection significantly contributes towards the success of the treatment.

  6. Clinical and molecular characteristics of Klebsiella pneumoniae ventilator-associated pneumonia in mainland China.

    PubMed

    Guo, Si; Xu, JingJing; Wei, YanShuan; Xu, JunHong; Li, Yi; Xue, Rui

    2016-10-26

    Klebsiella pneumoniae is a prominent nosocomial pathogen that accounts for up to 10 % of all hospital-acquired infections. It is a frequent cause of ventilator-associated pneumonia (VAP). The purpose of this study was to investigate the clinical characteristics of K. pneumoniae-associated VAP and the molecular characteristics of K. pneumoniae strains. We retrospectively reviewed 70 mechanically ventilated patients with K. pneumoniae isolated. All K. pneumoniae strains were examined to determine hypermucoviscosity (HV) phenotype, capsular serotypes, virulence genes, multilocus sequence typing and antimicrobial susceptibility. Hypermucoviscosity was found in 14 of 70 (20 %) isolates of K. pneumoniae. Among the 70 patients, 43 cases (61.4 %) developed VAP. Furthermore, VAP was more frequently induced by HV-positive K. pneumoniae (14/14, 100 %) than by HV-negative strains (29/56, 51.7 %). HV-positive K. pneumoniae-associated VAP patients were more inclined to develop bacteremia and had a higher mortality rate than HV-negative strains VAP patients. Antibiotic resistance was more frequent in HV- negative strains- than in HV- positive strains-infected patients. The prevalence of rmpA and aerobactin genes were 85.7 % and 85.7 % respectively, and serotypes K1 and K2 accounted for 14.3 % and 28.6 % of the hypermucoviscosity strains, respectively. Strains carrying rmpA and aerobactin genes were significantly associated with HV-phenotype, and rmpA and aerobactin coexisted in HV-positive strains. Multilocus sequence typing analysis identified 24 different sequence types from K. pneumoniae VAP samples. HV-phenotype is the major virulence determinant for mechanically ventilated patients. There was a specific sequence typing (ST) distribution between HV-positive and HV-negative strains.

  7. Klebsiella pneumoniae Siderophores Induce Inflammation, Bacterial Dissemination, and HIF-1α Stabilization during Pneumonia

    PubMed Central

    Holden, Victoria I.; Breen, Paul; Houle, Sébastien; Dozois, Charles M.

    2016-01-01

    ABSTRACT Klebsiella pneumoniae is a Gram-negative pathogen responsible for a wide range of infections, including pneumonia and bacteremia, and is rapidly acquiring antibiotic resistance. K. pneumoniae requires secretion of siderophores, low-molecular-weight, high-affinity iron chelators, for bacterial replication and full virulence. The specific combination of siderophores secreted by K. pneumoniae during infection can impact tissue localization, systemic dissemination, and host survival. However, the effect of these potent iron chelators on the host during infection is unknown. In vitro, siderophores deplete epithelial cell iron, induce cytokine secretion, and activate the master transcription factor hypoxia inducible factor-1α (HIF-1α) protein that controls vascular permeability and inflammatory gene expression. Therefore, we hypothesized that siderophore secretion by K. pneumoniae directly contributes to inflammation and bacterial dissemination during pneumonia. To examine the effects of siderophore secretion independently of bacterial growth, we performed infections with tonB mutants that persist in vivo but are deficient in siderophore import. Using a murine model of pneumonia, we found that siderophore secretion by K. pneumoniae induces the secretion of interleukin-6 (IL-6), CXCL1, and CXCL2, as well as bacterial dissemination to the spleen, compared to siderophore-negative mutants at an equivalent bacterial number. Furthermore, we determined that siderophore-secreting K. pneumoniae stabilized HIF-1α in vivo and that bacterial dissemination to the spleen required alveolar epithelial HIF-1α. Our results indicate that siderophores act directly on the host to induce inflammatory cytokines and bacterial dissemination and that HIF-1α is a susceptibility factor for bacterial invasion during pneumonia. PMID:27624128

  8. Interaction of lipocalin 2, transferrin, and siderophores determines the replicative niche of Klebsiella pneumoniae during pneumonia.

    PubMed

    Bachman, Michael A; Lenio, Steven; Schmidt, Lindsay; Oyler, Jennifer E; Weiser, Jeffrey N

    2012-11-20

    Pathogenic bacteria require iron for replication within their host. Klebsiella pneumoniae and other Gram-negative pathogens produce the prototypical siderophore enterobactin (Ent) to scavenge iron in vivo. In response, mucosal surfaces secrete lipocalin 2 (Lcn2), an innate immune protein that binds Ent to disrupt bacterial iron acquisition and promote acute inflammation during colonization. A subset of K. pneumoniae isolates attempt to evade Lcn2 by producing glycosylated Ent (Gly-Ent, salmochelin) or the alternative siderophore yersiniabactin (Ybt). However, these siderophores are not functionally equivalent and differ in their abilities to promote growth in the upper respiratory tract, lungs, and serum. To understand how Lcn2 exploits functional differences between siderophores, isogenic mutants of an Ent(+) Gly-Ent(+) Ybt(+) K. pneumoniae strain were inoculated into Lcn2(+/+) and Lcn2(-/-) mice, and the pattern of pneumonia was examined. Lcn2 effectively protected against the iroA ybtS mutant (Ent(+) Gly-Ent(-) Ybt(-)). Lcn2(+/+) mice had small foci of pneumonia, whereas Lcn2(-/-) mice had many bacteria in the perivascular space. The entB mutant (Ent(-) Ybt(+) Gly-Ent(-)) caused moderate bronchopneumonia but did not invade the transferrin-containing perivascular space. Accordingly, transferrin blocked Ybt-dependent growth in vitro. The wild type and the iroA mutant, which both produce Ent and Ybt, had a mixed phenotype, causing a moderate bronchopneumonia in Lcn2(+/+) mice and perivascular overgrowth in Lcn2(-/-) mice. Together, these data indicate that Lcn2, in combination with transferrin, confines K. pneumoniae to the airways and prevents invasion into tissue containing the pulmonary vasculature. Gram-negative bacteria are a common cause of severe hospital-acquired infections. To cause disease, they must obtain iron and secrete the small molecule enterobactin to do so. Animal models of pneumonia using Klebsiella pneumoniae indicate that enterobactin promotes

  9. Molecular epidemiology of carbapenem-resistant Klebsiella pneumoniae in Greece.

    PubMed

    Karampatakis, Theodoros; Antachopoulos, Charalampos; Iosifidis, Elias; Tsakris, Athanassios; Roilides, Emmanuel

    2016-06-01

    Hospital infections caused by carbapenem-resistant Klebsiella pneumoniae (CRKP) constitute a worldwide problem associated with high rates of treatment failure and mortality. In Greece, CRKP have emerged in 2002 due to VIM carbapenemase production and later due to KPC, NDM and OXA-48-like carbapenemases that have become endemic. The molecular epidemiology of CRKP strains is dynamic, as antibiotic consumption and worldwide traveling are strongly associated with global spread of CRKP isolates. Lately, porin defects, such as disruption of OmpK35 and production of OmpK36 variant, have also contributed to carbapenem resistance. In the coming years, the high prevalence of CRKP will require intense infection control measures, while novel molecular patterns may appear. To our knowledge, this is the first review analyzing the molecular epidemiology of CRKP strains in Greece.

  10. Prevalence of Extended Spectrum Beta-Lactamase Producing Klebsiella Pneumoniae Isolated From Urinary Tract Infected Patients.

    PubMed

    Chaudhary, P; Bhandari, D; Thapa, K; Thapa, P; Shrestha, D; Chaudhary, H K; Shrestha, A; Parajuli, H; Gupta, B P

    2016-05-01

    Klebsiella pneumoniae, one of the bacterial agents associated with urinary tract infection has been often implicated as a major extended spectrum beta-lactamase (ESBL) producer in last few decades. This study was designed to assess the prevalence of ESBL producing Klebsiella pneumoniae in urinary isolates at a tertiary care hospital in Kathmandu, Nepal, from July to December 2014. One thousand nine hundred eighty six mid-stream urine specimens were collected aseptically from the clinically suspected patients of urinary tract infections attending Capital Hospital and Research Center, Kathmandu. The samples were processed following standard guidelines as recommended by American Society for Microbiology (ASM) and the isolates including Klebsiella spp. were identified using the specific biochemical and sugar fermentation tests recommended by ASM. Antibiotic sensitivity testing was done by modified Kirby-Bauer disk diffusion method and interpreted following Clinical and Laboratory Standards Institute (CLSI) guidelines. Klebsiella pneumoniae isolates showing resistance upon initial screening with ceftriaxone (30 μg) disc were then confirmed for ESBL production by phenotypic confirmatory disc diffusion test (PCDDT) using ceftazidime (30 µg) and ceftazidime + clavulanic acid (30 µg + 10µg) and cefotaxime (30 µg) and cefotaxime + clavulanic acid (30 µg +10µg) disc as per CLSI guidelines. Out of a total 1986 specimens investigated, Escherichia coli was isolated in 309 (83.9%) and Klebsiella pneumoniae in 38 (10.3%) cases. Initial screening with ceftriaxone disc revealed 18 isolates of Klebsiella pneumoniae to be resistant. Further testing by PCDDT method confirmed 7 (18.4%) Klebsiella pneumoniae isolates to be ESBL producers. Compared to some earlier studies done in Nepal, higher prevalence of ESBL-producing Klebsiella pneumoniae was observed warranting a national surveillance for routine monitoring of ESBL producing Klebsiella pneumoniae isolates.

  11. Emergence of OXA-48-Producing Klebsiella pneumoniae in Taiwan.

    PubMed

    Ma, Ling; Wang, Jann-Tay; Wu, Tsu-Lan; Siu, L Kristopher; Chuang, Yin-Ching; Lin, Jung-Chung; Lu, Min-Chi; Lu, Po-Liang

    2015-01-01

    The isolation of OXA-48-producing Enterobacteriaceae has increased dramatically in Mediterranean countries in the past 10 years, and has recently emerged in Asia. Between January 2012 and May 2014, a total of 760 carbapenem non-susceptible Klebsiella pneumoniae (CnSKP) isolates were collected during a Taiwan national surveillance. Carbapenemases were detected in 210 CnSKP isolates (27.6%), including 162 KPC-2 (n = 1), KPC-3, KPC-17, and NDM-1 (n = 1 each), OXA-48 (n = 4), IMP-8 (n = 18), and VIM-1 (n = 24). The four blaOXA-48 CnSKP isolates were detected in late 2013. Herein we report the emergence OXA-48-producing K. pneumoniae isolates in Taiwan. PFGE analysis revealed that the four isolates belonged to three different pulsotypes. Three isolates harboured blaCTX-M genes and belonged to MLST type ST11. In addition, the plasmids belonged to the incompatibility group, IncA/C. One isolate belonged to ST116 and the plasmid incompatibility group was non-typeable. The sequence upstream of the blaOXA-48 gene in all four isolates was identical to pKPOXA-48N1, a blaOXA-48-carrying plasmid. This is the first report of OXA-48-producing Enterobacteriaceae in Taiwan and the second report to identify blaOXA-48 on an IncA/C plasmid in K. pneumoniae. Given that three isolates belong to the same pandemic clone (ST11) and possess the IncA/C plasmid and similar plasmid digestion profile that indicated the role of clonal spread or plasmid for dissemination of blaOXA-48 gene, the emergence of OXA-48-producing K. pneumoniae in Taiwan is of great concern.

  12. Nitrogen fixation by immobilized NIF derepressed Klebsiella pneumoniae cells

    SciTech Connect

    Venkatasubramanian, K.; Toda, Y.

    1980-01-01

    In vitro production of ammonia through biological means poses a number of challenges. The organisms should be able to accumulate considerable concentrations of ammonia in the medium. Secondly, nonphotosynthetic organisms must be supplied with high-energy substrates to carry out the fixation reaction. Thirdly, the organisms must be kept in a viable state to produce ammonia over long periods of time. In this article, preliminary results on the production of ammonia by a mutant strain of Klebsiella pneumoniae in continuous reactor systems are discussed. Continuous production of ammonia becomes feasible through the immobilization of the whole microbial cells and then through the use of the resulting catalyst system in a flow-through reactor. The rationale for immobilizing microbial cells and the advantages of such an approach over traditional fermentation processes are briefly described as they relate to the microbial production of ammonia. The microbial cells can be immobilized in such a way that their viability is still maintained in the immobilized state. This, in turn, obviates addition of cofactors, which is often an expensive step associated with immobilized multi-enzyme systems. Reconstituted bovine-hide collagen as the carrier matrix for fixing the cells was the carrier of choice for our work on immobilized Klebsiella cells. Polyacrylamide gels were examined as an alternate carrier matrix but results from this were found to be inferior to those collagen immobilized cell system.

  13. Klebsiella pneumoniae Siderophores Induce Inflammation, Bacterial Dissemination, and HIF-1α Stabilization during Pneumonia.

    PubMed

    Holden, Victoria I; Breen, Paul; Houle, Sébastien; Dozois, Charles M; Bachman, Michael A

    2016-09-13

    Klebsiella pneumoniae is a Gram-negative pathogen responsible for a wide range of infections, including pneumonia and bacteremia, and is rapidly acquiring antibiotic resistance. K. pneumoniae requires secretion of siderophores, low-molecular-weight, high-affinity iron chelators, for bacterial replication and full virulence. The specific combination of siderophores secreted by K. pneumoniae during infection can impact tissue localization, systemic dissemination, and host survival. However, the effect of these potent iron chelators on the host during infection is unknown. In vitro, siderophores deplete epithelial cell iron, induce cytokine secretion, and activate the master transcription factor hypoxia inducible factor-1α (HIF-1α) protein that controls vascular permeability and inflammatory gene expression. Therefore, we hypothesized that siderophore secretion by K. pneumoniae directly contributes to inflammation and bacterial dissemination during pneumonia. To examine the effects of siderophore secretion independently of bacterial growth, we performed infections with tonB mutants that persist in vivo but are deficient in siderophore import. Using a murine model of pneumonia, we found that siderophore secretion by K. pneumoniae induces the secretion of interleukin-6 (IL-6), CXCL1, and CXCL2, as well as bacterial dissemination to the spleen, compared to siderophore-negative mutants at an equivalent bacterial number. Furthermore, we determined that siderophore-secreting K. pneumoniae stabilized HIF-1α in vivo and that bacterial dissemination to the spleen required alveolar epithelial HIF-1α. Our results indicate that siderophores act directly on the host to induce inflammatory cytokines and bacterial dissemination and that HIF-1α is a susceptibility factor for bacterial invasion during pneumonia. Klebsiella pneumoniae causes a wide range of bacterial diseases, including pneumonia, urinary tract infections, and sepsis. To cause infection, K. pneumoniae steals

  14. Molecular characterization of multidrug-resistant Klebsiella pneumoniae isolates.

    PubMed

    Hou, Xiang-hua; Song, Xiu-yu; Ma, Xiao-bo; Zhang, Shi-yang; Zhang, Jia-qin

    2015-01-01

    Klebsiella pneumoniae is an important cause of healthcare-associated infections worldwide. Selective pressure, the extensive use of antibiotics, and the conjugational transmission of antibiotic resistance genes across bacterial species and genera facilitate the emergence of multidrug-resistant (MDR) K. pneumoniae. Here, we examined the occurrence, phenotypes and genetic features of MDR K. pneumoniae isolated from patients in intensive care units (ICUs) at the First Affiliated Hospital of Xiamen University in Xiamen, China, from January to December 2011. Thirty-eight MDR K. pneumoniae strains were collected. These MDR K. pneumoniae isolates possessed at least seven antibiotic resistance determinants, which contribute to the high-level resistance of these bacteria to aminoglycosides, macrolides, quinolones and β-lactams. Among these isolates, 24 strains were extended-spectrum β-lactamase (ESBL) producers, 2 strains were AmpC producers, and 12 strains were both ESBL and AmpC producers. The 38 MDR isolates also contained class I (28/38) and class II integrons (10/38). All 28 class I-positive isolates contained aacC1, aacC4, orfX, orfX' and aadA1 genes. β-lactam resistance was conferred through bla SHV (22/38), bla TEM (10/38), and bla CTX-M (7/38). The highly conserved bla KPC-2 (37/38) and bla OXA-23(1/38) alleles were responsible for carbapenem resistance, and a gyrAsite mutation (27/38) and the plasmid-mediated qnrB gene (13/38) were responsible for quinolone resistance. Repetitive-sequence-based PCR (REP-PCR) fingerprinting of these MDR strains revealed the presence of five groups and sixteen patterns. The MDR strains from unrelated groups showed different drug resistance patterns; however, some homologous strains also showed different drug resistance profiles. Therefore, REP-PCR-based analyses can provide information to evaluate the epidemic status of nosocomial infection caused by MDR K. pneumoniae; however, this test lacks the power to discriminate some

  15. Detection of drug-resistant Klebsiella pneumoniae in Chinese hares (Lepus sinensis).

    PubMed

    Du, Yingchun; Luo, Jing; Wang, Chengmin; Wen, Qingna; Duan, Mingxing; Zhang, Hong; He, Hongxuan

    2014-01-01

    We investigated an outbreak of acute pneumonia among adult Chinese hares (Lepus sinensis) and diarrhea among juvenile hares in Hebei Province, China, in 2012. Diagnosis was based on necropsy examination, microbial characteristics, biochemical identification, and nucleotide sequence analysis. The isolated bacteria from tissue samples of dead hares were identified as Klebsiella pneumoniae ssp. pneumoniae (K. pneumoniae). This K. pneumoniae was resistant to the antimicrobials imipenem, meropenem, penicillin, and vancomycin, but was highly sensitive to cefepime, cotrimoxazole, and enrofloxacin. Klebsiella pneumoniae is an important opportunistic pathogen, which often causes nosocomial infections in immunocompromised patients. However, the emergence of drug-resistant K. pneumoniae in hares indicates the existence of increasing risk of pathogen transmission between humans and wildlife. Given the close association between wildlife, livestock, and humans, it is important to identify epidemiologic factors associated with infection in these hares to minimize the risk of K. pneumoniae transmission.

  16. Presence of Nitrogen Fixing Klebsiella pneumoniae in the gut of the Formosan Subterranean Termite (Coptotermes formosanus)

    USDA-ARS?s Scientific Manuscript database

    A gram-negative facultative anaerobic enteric bacterium, Klebsiella pneumoniae was isolated from the hindgut of the Formosan subterranean termite (FST). It was characterized using, Fatty acid methyl ester (FAME) analysis, BIOLOG assay, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-...

  17. Klebsiella pneumoniae brain abscess in neonates: a report of 2 cases.

    PubMed

    Sundaram, Venkataseshan; Agrawal, Sunil; Chacham, Swathi; Mukhopadhyay, Kanya; Dutta, Sourabh; Kumar, Praveen

    2010-03-01

    Brain abscesses are uncommon in neonates. Klebsiella pneumoniae is a very uncommon microbial agent to cause brain abscess. We report 2 infants with Klebsiella pneumoniae sepsis who developed brain abscesses. One infant was a premature neonate who required mechanical ventilation for respiratory distress syndrome and subsequently developed nosocomial sepsis and brain abscess without evidence of preceding meningitis. Another infant was a full-term neonate without risk factors for sepsis who developed seizures on the sixth postnatal day and was found to have meningitis and brain abscess. Both infants had Klebsiella pneumoniae septicemia with multiple relatively large brain abscesses that responded poorly to antimicrobial agents. These infants were managed with transfontanel drainage and prolonged courses of antimicrobial agents. Key message of this report is that Klebsiella pneumoniae brain abscess may occur in the absence of meningitis and even in the absence of any identifiable risk factors.

  18. Complete Genome Sequence of Carbapenem-Resistant Klebsiella pneumoniae Strain 1756, Isolated from a Pus Specimen

    PubMed Central

    Kao, Cheng-Yen; Yan, Jing-Jou; Lin, Yu-Chun; Zheng, Po-Xing

    2017-01-01

    ABSTRACT Carbapenem-resistant Klebsiella pneumoniae strain 1756 was isolated from a pus specimen from a Taiwanese patient. Here, the complete genome sequence of strain 1756 is presented. PMID:28360152

  19. Rapidly fatal community-acquired pneumonia due to Klebsiella pneumoniae complicated with acute myocarditis and accelerated idioventricular rhythm.

    PubMed

    Chuang, Tzu-Yi; Lin, Chou-Jui; Lee, Shih-Wei; Chuang, Chun-Pin; Jong, Yuh-Shiun; Chen, Wen-Jone; Hsueh, Po-Ren

    2012-08-01

    We describe a previously healthy 52-year-old man with rapidly fatal community-acquired pneumonia caused by Klebsiella pneumoniae. The patient developed acute renal dysfunction, accelerated idioventricular rhythm (acute myocarditis), lactic acidosis and septic shock. He died within 15 hours after admission despite intravenous levofloxacin (750 mg daily) and aggressive medical treatment. Copyright © 2012. Published by Elsevier B.V.

  20. Nasopharyngeal carriage of Klebsiella pneumoniae and other Gram-negative bacilli in pneumonia-prone age groups in Semarang, Indonesia.

    PubMed

    Farida, Helmia; Severin, Juliëtte A; Gasem, M Hussein; Keuter, Monique; van den Broek, Peterhans; Hermans, Peter W M; Wahyono, Hendro; Verbrugh, Henri A

    2013-05-01

    Gram-negative bacilli (GNB) cause many cases of pneumonia in Indonesia. We investigated nasopharyngeal carriage of GNB in Semarang, Indonesia. Klebsiella pneumoniae carriage in adults (15%) was higher than in children (7%) (P = 0.004), while that of other GNB was comparable. Poor food and water hygiene are determinants of carriage of these bacteria.

  1. R-acetoin accumulation and dissimilation in Klebsiella pneumoniae.

    PubMed

    Wang, Dexin; Zhou, Jidong; Chen, Chuan; Wei, Dong; Shi, Jiping; Jiang, Biao; Liu, Pengfu; Hao, Jian

    2015-08-01

    Klebsiella pneumoniae is a 2,3-butanediol producer, and R-acetoin is an intermediate of 2,3-butanediol production. R-acetoin accumulation and dissimilation in K. pneumoniae was studied here. A budC mutant, which has lost 2,3-butanediol dehydrogenase activity, accumulated high levels of R-acetoin in culture broth. However, after glucose was exhausted, the accumulated R-acetoin could be reused by the cells as a carbon source. Acetoin dehydrogenase enzyme system, encoded by acoABCD, was responsible for R-acetoin dissimilation. acoABCD mutants lost the ability to grow on acetoin as the sole carbon source, and the acetoin accumulated could not be dissimilated. However, in the presence of another carbon source, the acetoin accumulated in broth of acoABCD mutants was converted to 2,3-butanediol. Parameters of R-acetoin production by budC mutants were optimized in batch culture. Aerobic culture and mildly acidic conditions (pH 6-6.5) favored R-acetoin accumulation. At the optimized conditions, in fed-batch fermentation, 62.3 g/L R-acetoin was produced by budC and acoABCD double mutant in 57 h culture, with an optical purity of 98.0 %, and a substrate conversion ratio of 28.7 %.

  2. The secondary resistome of multidrug-resistant Klebsiella pneumoniae

    PubMed Central

    Jana, Bimal; Cain, Amy K.; Doerrler, William T.; Boinett, Christine J.; Fookes, Maria C.; Parkhill, Julian; Guardabassi, Luca

    2017-01-01

    Klebsiella pneumoniae causes severe lung and bloodstream infections that are difficult to treat due to multidrug resistance. We hypothesized that antimicrobial resistance can be reversed by targeting chromosomal non-essential genes that are not responsible for acquired resistance but essential for resistant bacteria under therapeutic concentrations of antimicrobials. Conditional essentiality of individual genes to antimicrobial resistance was evaluated in an epidemic multidrug-resistant clone of K. pneumoniae (ST258). We constructed a high-density transposon mutant library of >430,000 unique Tn5 insertions and measured mutant depletion upon exposure to three clinically relevant antimicrobials (colistin, imipenem or ciprofloxacin) by Transposon Directed Insertion-site Sequencing (TraDIS). Using this high-throughput approach, we defined three sets of chromosomal non-essential genes essential for growth during exposure to colistin (n = 35), imipenem (n = 1) or ciprofloxacin (n = 1) in addition to known resistance determinants, collectively termed the “secondary resistome”. As proof of principle, we demonstrated that inactivation of a non-essential gene not previously found linked to colistin resistance (dedA) restored colistin susceptibility by reducing the minimum inhibitory concentration from 8 to 0.5 μg/ml, 4-fold below the susceptibility breakpoint (S ≤ 2 μg/ml). This finding suggests that the secondary resistome is a potential target for developing antimicrobial “helper” drugs that restore the efficacy of existing antimicrobials. PMID:28198411

  3. The secondary resistome of multidrug-resistant Klebsiella pneumoniae.

    PubMed

    Jana, Bimal; Cain, Amy K; Doerrler, William T; Boinett, Christine J; Fookes, Maria C; Parkhill, Julian; Guardabassi, Luca

    2017-02-15

    Klebsiella pneumoniae causes severe lung and bloodstream infections that are difficult to treat due to multidrug resistance. We hypothesized that antimicrobial resistance can be reversed by targeting chromosomal non-essential genes that are not responsible for acquired resistance but essential for resistant bacteria under therapeutic concentrations of antimicrobials. Conditional essentiality of individual genes to antimicrobial resistance was evaluated in an epidemic multidrug-resistant clone of K. pneumoniae (ST258). We constructed a high-density transposon mutant library of >430,000 unique Tn5 insertions and measured mutant depletion upon exposure to three clinically relevant antimicrobials (colistin, imipenem or ciprofloxacin) by Transposon Directed Insertion-site Sequencing (TraDIS). Using this high-throughput approach, we defined three sets of chromosomal non-essential genes essential for growth during exposure to colistin (n = 35), imipenem (n = 1) or ciprofloxacin (n = 1) in addition to known resistance determinants, collectively termed the "secondary resistome". As proof of principle, we demonstrated that inactivation of a non-essential gene not previously found linked to colistin resistance (dedA) restored colistin susceptibility by reducing the minimum inhibitory concentration from 8 to 0.5 μg/ml, 4-fold below the susceptibility breakpoint (S ≤ 2 μg/ml). This finding suggests that the secondary resistome is a potential target for developing antimicrobial "helper" drugs that restore the efficacy of existing antimicrobials.

  4. Rapid susceptibility profiling of carbapenem-resistant Klebsiella pneumoniae.

    PubMed

    Mulroney, K T; Hall, J M; Huang, X; Turnbull, E; Bzdyl, N M; Chakera, A; Naseer, U; Corea, E M; Ellington, M J; Hopkins, K L; Wester, A L; Ekelund, O; Woodford, N; Inglis, T J J

    2017-05-15

    The expanding global distribution of multi-resistant Klebsiella pneumoniae demands faster antimicrobial susceptibility testing (AST) to guide antibiotic treatment. Current ASTs rely on time-consuming differentiation of resistance and susceptibility after initial isolation of bacteria from a clinical specimen. Here we describe a flow cytometry workflow to determine carbapenem susceptibility from bacterial cell characteristics in an international K. pneumoniae isolate collection (n = 48), with a range of carbapenemases. Our flow cytometry-assisted susceptibility test (FAST) method combines rapid qualitative susceptible/non-susceptible classification and quantitative MIC measurement in a single process completed shortly after receipt of a primary isolate (54 and 158 minutes respectively). The qualitative FAST results and FAST-derived MIC (MICFAST) correspond closely with broth microdilution MIC (MICBMD, Matthew's correlation coefficient 0.887), align with the international AST standard (ISO 200776-1; 2006) and could be used for rapid determination of antimicrobial susceptibility in a wider range of Gram negative and Gram positive bacteria.

  5. CTX-M-12 β-Lactamase in a Klebsiella pneumoniae Clinical Isolate in Colombia

    PubMed Central

    Villegas, Maria Virginia; Correa, Adriana; Perez, Federico; Zuluaga, Tania; Radice, Marcela; Gutkind, Gabriel; Casellas, José María; Ayala, Juan; Lolans, Karen; Quinn, John P.

    2004-01-01

    We describe the detection of the CTX-M-12 β-lactamase from a clinical isolate of Klebsiella pneumoniae in Colombia. Screening of nosocomial Klebsiella spp. and Escherichia coli isolates from a network of teaching hospitals revealed the presence of CTX-M enzymes in multiple cities. This is the first description of CTX-M in Colombia. PMID:14742223

  6. CTX-M-12 beta-lactamase in a Klebsiella pneumoniae clinical isolate in Colombia.

    PubMed

    Villegas, Maria Virginia; Correa, Adriana; Perez, Federico; Zuluaga, Tania; Radice, Marcela; Gutkind, Gabriel; Casellas, José María; Ayala, Juan; Lolans, Karen; Quinn, John P

    2004-02-01

    We describe the detection of the CTX-M-12 beta-lactamase from a clinical isolate of Klebsiella pneumoniae in Colombia. Screening of nosocomial Klebsiella spp. and Escherichia coli isolates from a network of teaching hospitals revealed the presence of CTX-M enzymes in multiple cities. This is the first description of CTX-M in Colombia.

  7. Genome Sequence of Klebsiella pneumoniae Bacteriophage PMBT1 Isolated from Raw Sewage

    PubMed Central

    Brinks, Erik; Fiedler, Gregor; Hüsing, Christina; Cho, Gyu-Sung; Hoeppner, Marc P.; Heller, Knut J.; Neve, Horst; Franz, Charles M. A. P.

    2017-01-01

    ABSTRACT A bacteriophage virulent for extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae strain 182 was isolated from sewage. The double-stranded DNA (dsDNA) genome showed high similarity to the genomes of other Klebsiella pneumoniae phages. It comprises 175,206 bp with a mol% G+C content of 41.9 and contains 276 putative open reading frames (ORFs) and one tRNA. PMID:28232430

  8. Molecular Epidemiological Characteristics of Klebsiella pneumoniae Associated with Bacteremia among Patients with Pneumonia

    PubMed Central

    Ito, Ryota; Shindo, Yuichiro; Kobayashi, Daisuke; Ando, Masahiko; Jin, Wanchun; Wachino, Jun-ichi; Yamada, Keiko; Kimura, Kouji; Yagi, Tetsuya; Hasegawa, Yoshinori

    2015-01-01

    Some important virulence factors have been elucidated in Klebsiella pneumoniae infections. We investigated the relationship between virulence factors and multilocus sequence types (STs) and assessed the risk factors for bacteremia in patients with pneumonia due to K. pneumoniae. From April 2004 through April 2012, a total of 120 K. pneumoniae isolates from patients with pneumonia (23 with bacteremia and 97 without bacteremia) were collected from 10 medical institutions in Japan. Additionally, 10 strains of K. pneumoniae serotype K2 that were isolated >30 years ago were included in this study. These isolates were characterized using multilocus sequence typing (MLST), and the characteristics of their virulence factors, such as hypermucoviscosity phenotype and RmpA and aerobactin production between patients with and without bacteremia, were examined. MLST analysis was performed on the 120 isolates from patients with pneumonia, and some sequence type groups were defined as genetic lineages (GLs). GL65 was more prevalent among patients with bacteremia (21.7%) than in those without bacteremia (7.2%). The majority of the strains with serotype K2 were classified into GL14 or GL65, and rmpA and the gene for aerobactin were present in all GL65-K2 strains but absent in all GL14-K2 strains. In a multivariate analysis, the independent risk factors for bacteremia included GL65 (adjusted odds ratio [AOR], 9.46; 95% confidence interval [CI], 1.81 to 49.31), as well as neoplastic disease (AOR, 9.94; 95% CI, 2.61 to 37.92), immunosuppression (AOR, 17.85; 95% CI, 1.49 to 214.17), and hypoalbuminemia (AOR, 4.76; 95% CI, 1.29 to 17.61). GL65 was more prevalent among patients with bacteremia and was associated with the virulence factors of K. pneumoniae. PMID:25568434

  9. Molecular epidemiological characteristics of Klebsiella pneumoniae associated with bacteremia among patients with pneumonia.

    PubMed

    Ito, Ryota; Shindo, Yuichiro; Kobayashi, Daisuke; Ando, Masahiko; Jin, Wanchun; Wachino, Jun-ichi; Yamada, Keiko; Kimura, Kouji; Yagi, Tetsuya; Hasegawa, Yoshinori; Arakawa, Yoshichika

    2015-03-01

    Some important virulence factors have been elucidated in Klebsiella pneumoniae infections. We investigated the relationship between virulence factors and multilocus sequence types (STs) and assessed the risk factors for bacteremia in patients with pneumonia due to K. pneumoniae. From April 2004 through April 2012, a total of 120 K. pneumoniae isolates from patients with pneumonia (23 with bacteremia and 97 without bacteremia) were collected from 10 medical institutions in Japan. Additionally, 10 strains of K. pneumoniae serotype K2 that were isolated >30 years ago were included in this study. These isolates were characterized using multilocus sequence typing (MLST), and the characteristics of their virulence factors, such as hypermucoviscosity phenotype and RmpA and aerobactin production between patients with and without bacteremia, were examined. MLST analysis was performed on the 120 isolates from patients with pneumonia, and some sequence type groups were defined as genetic lineages (GLs). GL65 was more prevalent among patients with bacteremia (21.7%) than in those without bacteremia (7.2%). The majority of the strains with serotype K2 were classified into GL14 or GL65, and rmpA and the gene for aerobactin were present in all GL65-K2 strains but absent in all GL14-K2 strains. In a multivariate analysis, the independent risk factors for bacteremia included GL65 (adjusted odds ratio [AOR], 9.46; 95% confidence interval [CI], 1.81 to 49.31), as well as neoplastic disease (AOR, 9.94; 95% CI, 2.61 to 37.92), immunosuppression (AOR, 17.85; 95% CI, 1.49 to 214.17), and hypoalbuminemia (AOR, 4.76; 95% CI, 1.29 to 17.61). GL65 was more prevalent among patients with bacteremia and was associated with the virulence factors of K. pneumoniae. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  10. Klebsiella pneumoniae as a cause of pneumonia and septicemia in a civet kitten (Civettictis civetta) in the Jos Zoo, Nigeria.

    PubMed

    Enurah, L U; Adeniyi, K O; Ocholi, R A; Spencer, T H; Badung, J D

    1988-07-01

    A fatal case of acute pneumonia and septicemia that occurred in a captive civet kitten (Civettictis civetta) in the Jos Zoo, Nigeria is reported. Diagnosis was based on clinical signs, necropsy findings, and the isolation of Klebsiella pneumoniae from the lung, intestine, liver and heart blood of the animal. This is the first report of clinical K. pneumoniae infection in a wild or captive animal in Nigeria.

  11. Klebsiella pneumoniae Produces No Histamine: Raoultella planticola and Raoultella ornithinolytica Strains Are Histamine Producers

    PubMed Central

    Kanki, Masashi; Yoda, Tomoko; Tsukamoto, Teizo; Shibata, Tadayoshi

    2002-01-01

    Histamine fish poisoning is caused by histamine-producing bacteria (HPB). Klebsiella pneumoniae and Klebsiella oxytoca are the best-known HPB in fish. However, 22 strains of HPB from fish first identified as K. pneumoniae or K. oxytoca by commercialized systems were later correctly identified as Raoultella planticola (formerly Klebsiella planticola) by additional tests. Similarly, five strains of Raoultella ornithinolytica (formerly Klebsiella ornithinolytica) were isolated from fish as new HPB. R. planticola and R. ornithinolytica strains were equal in their histamine-producing capabilities and were determined to possess the hdc genes, encoding histidine decarboxylase. On the other hand, a collection of 61 strains of K. pneumoniae and 18 strains of K. oxytoca produced no histamine. PMID:12089029

  12. Draft Genome Sequence of a Klebsiella pneumoniae Strain (New Sequence Type 2357) Carrying Tn3926

    PubMed Central

    Mi, Zu-huang; Wang, Chun-xin; Zhu, Jian-ming

    2016-01-01

    We present the draft genome sequence of a Klebsiella pneumoniae carbapenemase–producing sequence type 2357 (ST2357) strain, NB60, which contains drug-resistant genes encoding resistance to beta-lactams, fluoroquinolones, aminoglycosides, trimethoprim-sulfamethoxazole, colistin, macrolides, and tetracycline. Strain NB60 was isolated from human blood, making it an important tool for studying K. pneumoniae pathogenesis. PMID:27660779

  13. Complete Genome Sequence of KPC-Producing Klebsiella pneumoniae Strain CAV1193.

    PubMed

    Sheppard, Anna E; Stoesser, Nicole; Sebra, Robert; Kasarskis, Andrew; Deikus, Gintaras; Anson, Luke; Walker, A Sarah; Peto, Tim E; Crook, Derrick W; Mathers, Amy J

    2016-01-28

    Carbapenem resistance in Klebsiella pneumoniae, frequently conferred by the blaKPC gene, is a major public health threat. We sequenced a blaKPC-containing strain of K. pneumoniae belonging to the emergent lineage ST941, in order to better understand the evolution of blaKPC within this species. Copyright © 2016 Sheppard et al.

  14. Draft Genome Sequences of Three Hypervirulent Carbapenem-Resistant Klebsiella pneumoniae Isolates from Bacteremia

    PubMed Central

    Shankar, Chaitra; Nabarro, Laura E. B.; Devanga Ragupathi, Naveen Kumar; Muthuirulandi Sethuvel, Dhiviya Prabaa; Daniel, Jones Lionel Kumar; Doss C, George Priya

    2016-01-01

    Hypervirulent Klebsiella pneumoniae strains have been increasingly reported worldwide, and there is emergence of carbapenem resistance among them. Here, we report the genome sequences of three carbapenem-resistant hypervirulent K. pneumoniae isolates isolated from bacteremic patients at a tertiary-care center in South India. PMID:27932638

  15. Complete Genome Sequence of KPC-Producing Klebsiella pneumoniae Strain CAV1193

    PubMed Central

    Sebra, Robert; Kasarskis, Andrew; Deikus, Gintaras; Anson, Luke; Walker, A. Sarah; Peto, Tim E.; Crook, Derrick W.

    2016-01-01

    Carbapenem resistance in Klebsiella pneumoniae, frequently conferred by the blaKPC gene, is a major public health threat. We sequenced a blaKPC-containing strain of K. pneumoniae belonging to the emergent lineage ST941, in order to better understand the evolution of blaKPC within this species. PMID:26823590

  16. Phenotypic and genotypic characterization of Klebsiella pneumonia recovered from nonhuman primates

    USDA-ARS?s Scientific Manuscript database

    Klebsiella pneumoniae is a zoonotic, Gram-negative member of the family Enterobacteriaceae and is the causative agent of nosocomial septicemic, pneumonic, and urinary tract infections. Recently, pathogenic strains of K. pneumoniae sharing a hypermucoviscosity (HMV) phenotype have been attributed to ...

  17. Predatory Bacteria Attenuate Klebsiella pneumoniae Burden in Rat Lungs

    PubMed Central

    Singleton, Eric; Tang, Chi; Zuena, Michael; Shukla, Sean; Gupta, Shilpi; Dharani, Sonal; Onyile, Onoyom; Rinaggio, Joseph; Connell, Nancy D.

    2016-01-01

    ABSTRACT Bdellovibrio bacteriovorus and Micavibrio aeruginosavorus are predatory bacteria that naturally—and obligately—prey on other Gram-negative bacteria, and their use has been proposed as a potential new approach to control microbial infection. The ability of predatory bacteria to prey on Gram-negative human pathogens in vitro is well documented; however, the in vivo safety and efficacy of predatory bacteria have yet to be fully assessed. In this study, we examined whether predatory bacteria can reduce bacterial burden in the lungs in an in vivo mammalian system. Initial safety studies were performed by intranasal inoculation of rats with predatory bacteria. No adverse effects or lung pathology were observed in rats exposed to high concentrations of predatory bacteria at up to 10 days postinoculation. Enzyme-linked immunosorbent assay (ELISA) of the immune response revealed a slight increase in inflammatory cytokine levels at 1 h postinoculation that was not sustained by 48 h. Additionally, dissemination experiments showed that predators were efficiently cleared from the host by 10 days postinoculation. To measure the ability of predatory bacteria to reduce microbial burden in vivo, we introduced sublethal concentrations of Klebsiella pneumoniae into the lungs of rats via intranasal inoculation and followed with multiple doses of predatory bacteria over 24 h. Predatory bacteria were able to reduce K. pneumoniae bacterial burden, on average, by more than 3.0 log10 in the lungs of most rats as measured by CFU plating. The work presented here provides further support for the idea of developing predatory bacteria as a novel biocontrol agent. PMID:27834203

  18. Klebsiella pneumoniae secretes outer membrane vesicles that induce the innate immune response.

    PubMed

    Lee, Je Chul; Lee, Eun Jeoung; Lee, Jung Hwa; Jun, So Hyun; Choi, Chi Won; Kim, Seung Il; Kang, Sang Sun; Hyun, Sunghee

    2012-06-01

    Outer membrane vesicles (OMVs) derived from pathogenic Gram-negative bacteria are an important vehicle for delivery of effector molecules to host cells, but the production of OMVs from Klebsiella pneumoniae, an opportunistic pathogen of both nosocomial and community-acquired infections, and their role in bacterial pathogenesis have not yet been determined. In the present study, we examined the production of OMVs from K. pneumoniae and determined the induction of the innate immune response against K. pneumoniae OMVs. Klebsiella pneumoniae ATCC 13883 produced and secreted OMVs during in vitro culture. Proteomic analysis revealed that 159 different proteins were associated with K. pneumoniae OMVs. Klebsiella pneumoniae OMVs did not inhibit cell growth or induce cell death. However, these vesicles induced expression of proinflammatory cytokine genes such as interleukin (IL)-1β and IL-8 in epithelial cells. An intratracheal challenge of K. pneumoniae OMVs in neutropenic mice resulted in severe lung pathology similar to K. pneumoniae infection. In conclusion, K. pneumoniae produces OMVs like other pathogenic Gram-negative bacteria and K. pneumoniae OMVs are a molecular complex that induces the innate immune response.

  19. Conventional NK cells can produce IL-22 and promote host defense in Klebsiella pneumoniae pneumonia.

    PubMed

    Xu, Xin; Weiss, Ido D; Zhang, Hongwei H; Singh, Satya P; Wynn, Thomas A; Wilson, Mark S; Farber, Joshua M

    2014-02-15

    It was reported that host defense against pulmonary Klebsiella pneumoniae infection requires IL-22, which was proposed to be of T cell origin. Supporting a role for IL-22, we found that Il22(-/-) mice had decreased survival compared with wild-type mice after intratracheal infection with K. pneumoniae. Surprisingly, however, Rag2(-/-) mice did not differ from wild-type mice in survival or levels of IL-22 in the lungs postinfection with K. pneumoniae. In contrast, K. pneumoniae-infected Rag2(-/-)Il2rg(-/-) mice failed to produce IL-22. These data suggested a possible role for NK cells or other innate lymphoid cells in host defense and production of IL-22. Unlike NK cell-like innate lymphoid cells that produce IL-22 and display a surface phenotype of NK1.1(-)NKp46(+)CCR6(+), lung NK cells showed the conventional phenotype, NK1.1(+)NKp46(+)CCR6(-). Mice depleted of NK cells using anti-asialo GM1 showed decreased survival and higher lung bacterial counts, as well as increased dissemination of K. pneumoniae to blood and liver, compared with control-treated mice. NK cell depletion also led to decreased production of IL-22 in the lung. Within 1 d postinfection, although there was no increase in the number of lung NK cells, a subset of lung NK cells became competent to produce IL-22, and such cells were found in both wild-type and Rag2(-/-) mice. Our data suggest that, during pulmonary infection of mice with K. pneumoniae, conventional NK cells are required for optimal host defense, which includes the production of IL-22.

  20. Interaction of Lipocalin 2, Transferrin, and Siderophores Determines the Replicative Niche of Klebsiella pneumoniae during Pneumonia

    PubMed Central

    Bachman, Michael A.; Lenio, Steven; Schmidt, Lindsay; Oyler, Jennifer E.; Weiser, Jeffrey N.

    2012-01-01

    ABSTRACT Pathogenic bacteria require iron for replication within their host. Klebsiella pneumoniae and other Gram-negative pathogens produce the prototypical siderophore enterobactin (Ent) to scavenge iron in vivo. In response, mucosal surfaces secrete lipocalin 2 (Lcn2), an innate immune protein that binds Ent to disrupt bacterial iron acquisition and promote acute inflammation during colonization. A subset of K. pneumoniae isolates attempt to evade Lcn2 by producing glycosylated Ent (Gly-Ent, salmochelin) or the alternative siderophore yersiniabactin (Ybt). However, these siderophores are not functionally equivalent and differ in their abilities to promote growth in the upper respiratory tract, lungs, and serum. To understand how Lcn2 exploits functional differences between siderophores, isogenic mutants of an Ent+ Gly-Ent+ Ybt+ K. pneumoniae strain were inoculated into Lcn2+/+ and Lcn2−/− mice, and the pattern of pneumonia was examined. Lcn2 effectively protected against the iroA ybtS mutant (Ent+ Gly-Ent− Ybt−). Lcn2+/+ mice had small foci of pneumonia, whereas Lcn2−/− mice had many bacteria in the perivascular space. The entB mutant (Ent− Ybt+ Gly-Ent−) caused moderate bronchopneumonia but did not invade the transferrin-containing perivascular space. Accordingly, transferrin blocked Ybt-dependent growth in vitro. The wild type and the iroA mutant, which both produce Ent and Ybt, had a mixed phenotype, causing a moderate bronchopneumonia in Lcn2+/+ mice and perivascular overgrowth in Lcn2−/− mice. Together, these data indicate that Lcn2, in combination with transferrin, confines K. pneumoniae to the airways and prevents invasion into tissue containing the pulmonary vasculature. PMID:23169997

  1. Klebsiella pneumoniae 1,3-propanediol:NAD+ oxidoreductase.

    PubMed Central

    Johnson, E A; Lin, E C

    1987-01-01

    Fermentative utilization of glycerol, a more reduced carbohydrate than aldoses and ketoses, requires the disposal of the two extra hydrogen atoms. This is accomplished by sacrificing an equal quantity of glycerol via an auxiliary pathway initiated by glycerol dehydratase. The product, 3-hydroxypropionaldehyde, is then reduced by 1,3-propanediol NAD+:oxidoreductase (1,3-propanediol dehydrogenase; EC 1.1.1.202), resulting in the regeneration of NAD+ from NADH. The pathway for the assimilation of glycerol is initiated by an NAD-linked dehydrogenase. In Klebsiella pneumoniae the two pathways are encoded by the dha regulon which is inducible only anaerobically. In this study 1,3-propanediol:NAD+ oxidoreductase was purified from cells grown anaerobically on glycerol. The enzyme was immunochemically distinct from the NAD-linked glycerol dehydrogenase and was an octamer or hexamer of a polypeptide of 45,000 +/- 3,000 daltons. When tested as a dehydrogenase, only 1,3-propanediol served as a substrate; no activity was detected with ethanol, 1-propanol, 1,2-propanediol, glycerol, or 1,4-butanediol. The enzyme was inhibited by chelators of divalent cations. An enzyme preparation inhibited by alpha,alpha'-dipyridyl was reactivated by the addition of Fe2+ or Mn2+ after removal of the chelator by gel filtration. As for glycerol dehydrogenase, 1,3-propanediol oxidoreductase is apparently inactivated by oxidation during aerobic metabolism, under which condition the enzyme becomes superfluous. Images PMID:3553154

  2. [KPC-producing Klebsiella pneumoniae colonization at a University Hospital].

    PubMed

    Echavarría, Gonzalo L; Guevara Nuñez, Daiana; Bertona, Eugenia; De Paulis, Adriana N; Predari, Silvia C; Benchetrit, Guillermo

    2017-01-01

    A surveillance study was conducted at a University Hospital in Buenos Aires City aimed to assess the rates of colonization with carbapenemase-producing strains of Klebsiella pneumoniae, which are bacteria of utmost epidemiological importance. To this end, rectal swabs collected from all inpatients were cultured for the presence of these bacteria. Two point prevalence surveys showed high prevalence rates (up to 25%). The following variables were evaluated in all inpatients: place of origin (home or other chronic care center), age, prolonged hospitalization, antibiotics for at least 72 hours prior to swabbing, intensive care unit requirements for at least 24 hours, mechanical ventilation assistance for more than 4 days, hemodialysis requirements, need for surgery, enteral feeding through a nasogastric tube, and functional evaluation according to the Karnofsky performance scale. The variable associated with the highest statistical significance was the use of nasogastric enteral feeding. Also, the length of stay was significantly higher and the functional status was significantly worse in colonized patients. As for the prior use of antibiotics, results were close to statistical significance but without reaching it. Measures were implemented in order to control the spread of the microorganism in the acute setting and beyond. Upon implementation of such measures, a third prevalence survey was performed that showed a decrease in the horizontal transmission of the microorganism.

  3. Structure of 2-oxo-3-deoxygalactonate kinase from Klebsiella pneumoniae

    PubMed Central

    Michalska, Karolina; Cuff, Marianne E.; Tesar, Christine; Feldmann, Brian; Joachimiak, Andrzej

    2011-01-01

    In most organisms, efficient d-galactose utilization requires the highly conserved Leloir pathway that converts d-galactose to d-glucose 1-phosphate. However, in some bacterial and fungal species alternative routes of d-galactose assimilation have been identified. In the so-called De Ley–Doudoroff pathway, d-galactose is metabolized into pyruvate and d-­glyceraldehyde 3-phosphate in five consecutive reactions carried out by specific enzymes. The penultimate step in this pathway involves the phosphorylation of 2-oxo-3-deoxygalactonate to 2-oxo-3-deoxygalactonate 6-phosphate catalyzed by 2-­oxo-3-deoxygalactonate kinase, with ATP serving as a phosphoryl-group donor. Here, a crystal structure of 2-oxo-3-deoxygalactonate kinase from Klebsiella pneumoniae determined at 2.1 Å resolution is reported, the first structure of an enzyme from the De Ley–Doudoroff pathway. Structural comparison indicates that the enzyme belongs to the ASKHA (acetate and sugar kinases/hsc70/actin) family of phosphotransferases. The protein is composed of two α/β domains, each of which contains a core common to all family members. Additional elements introduced between conserved structural motifs define the unique features of 2-oxo-3-deoxygalactonate kinase and possibly determine the biological function of the protein. PMID:21795809

  4. Structure of 2-oxo-3-deoxygalactonate kinase from Klebsiella pneumoniae

    SciTech Connect

    Michalska, Karolina; Cuff, Marianne E.; Tesar, Christine; Feldmann, Brian; Joachimiak, Andrzej

    2011-08-01

    The crystal structure of 2-oxo-3-deoxygalactonate kinase from the De Ley–Doudoroff pathway of galactose metabolism has been determined at 2.1 Å resolution. In most organisms, efficient d-galactose utilization requires the highly conserved Leloir pathway that converts d-galactose to d-glucose 1-phosphate. However, in some bacterial and fungal species alternative routes of d-galactose assimilation have been identified. In the so-called De Ley–Doudoroff pathway, d-galactose is metabolized into pyruvate and d-glyceraldehyde 3-phosphate in five consecutive reactions carried out by specific enzymes. The penultimate step in this pathway involves the phosphorylation of 2-oxo-3-deoxygalactonate to 2-oxo-3-deoxygalactonate 6-phosphate catalyzed by 2-oxo-3-deoxygalactonate kinase, with ATP serving as a phosphoryl-group donor. Here, a crystal structure of 2-oxo-3-deoxygalactonate kinase from Klebsiella pneumoniae determined at 2.1 Å resolution is reported, the first structure of an enzyme from the De Ley–Doudoroff pathway. Structural comparison indicates that the enzyme belongs to the ASKHA (acetate and sugar kinases/hsc70/actin) family of phosphotransferases. The protein is composed of two α/β domains, each of which contains a core common to all family members. Additional elements introduced between conserved structural motifs define the unique features of 2-oxo-3-deoxygalactonate kinase and possibly determine the biological function of the protein.

  5. Community-Acquired Klebsiella pneumoniae Bacteremia: Global Differences in Clinical Patterns

    PubMed Central

    Ko, Wen-Chien; Paterson, David L.; Sagnimeni, Anthanasia J.; Hansen, Dennis S.; Von Gottberg, Anne; Mohapatra, Sunita; Casellas, Jose Maria; Goossens, Herman; Mulazimoglu, Lutfiye; Trenholme, Gordon; Klugman, Keith P.; McCormack, Joseph G.

    2002-01-01

    We initiated a worldwide collaborative study, including 455 episodes of bacteremia, to elucidate the clinical patterns of Klebsiella pneumoniae. Historically, community-acquired pneumonia has been consistently associated with K. pneumoniae. Only four cases of community-acquired bacteremic K. pneumoniae pneumonia were seen in the 2-year study period in the United States, Argentina, Europe, or Australia; none were in alcoholics. In contrast, 53 cases of bacteremic K. pneumoniae pneumonia were observed in South Africa and Taiwan, where an association with alcoholism persisted (p=0.007). Twenty-five cases of a distinctive syndrome consisting of K. pneumoniae bacteremia in conjunction with community-acquired liver abscess, meningitis, or endophthalmitis were observed. A distinctive form of K. pneumoniae infection, often causing liver abscess, was identified, almost exclusively in Taiwan. PMID:11897067

  6. [Relationship between isolation of extended spectrum beta-lactamase-producing Klebsiella pneumoniae and course of hospital-acquired pneumonia].

    PubMed

    Vitkauskiene, Astra; Giedraitiene, Agne; Dudzevicius, Vytis; Sakalauskas, Raimundas

    2007-01-01

    To evaluate relationship between isolation of extended spectrum beta-lactamase-producing Klebsiella pneumoniae strains and course of hospital-acquired pneumonia. K. pneumoniae strains isolated from bronchial secretions or bronchoalveolar lavage fluid samples of patients hospitalized at an intensive care unit of Kaunas University of Medicine Hospital were analyzed. By means of synergistic two-antibiotics disc method, K. pneumoniae strains producing extended spectrum beta-lactamases were selected for further analysis using E-test (AB Biodisk, Solna, Sweden). Hospital-acquired pneumonia was diagnosed based on standard criteria for the diagnosis of pneumonia if signs of pneumonia occurred after 48 hours following admission. Late-onset hospital-acquired pneumonia was considered if these signs of pneumonia occurred on fifth day of hospitalization or later. Total of 45 strains of K. pneumoniae were isolated during the study period; 18 isolated strains produced ESBL. Thirty-two patients investigated have developed hospital-acquired pneumonia, 20 of which were cases of late-onset hospital-acquired pneumonia. Thirteen cases of K. pneumoniae isolation were classified as airway colonization. Extended spectrum beta-lactamase-producing K. pneumoniae strains were more frequently isolated from patients with hospital-acquired pneumonia (88.9%, n=16 and 11.1%, n=2, P<0.05) in comparison with non-producing strains. Extended spectrum beta-lactamase-producing strains were more prevalent in late-onset pneumonia group (93.8%, n=15) than in early-onset group (6.2%, n=1, P<0.001). Extended spectrum beta-lactamase-producing K. pneumoniae strains were more frequently isolated from patients with hospital-acquired pneumonia as compared to colonized patients. Extended spectrum beta-lactamase-producing K. pneumoniae strains were more frequently isolated from patients with late-onset hospital-acquired pneumonia.

  7. Factors influencing inactivation of Klebsiella pneumoniae by chlorine and chloramine.

    PubMed

    Goel, Sudha; Bouwer, Edward J

    2004-01-01

    Inactivation of Klebsiella pneumoniae cultures by chlorine and chloramine was evaluated under different growth conditions by varying nutrient media dilution, concentrations of essential inorganic nutrients (FeCl3, MgSO4, phosphate, and ammonium salts), and temperature. All inactivation assays were performed at room temperature (22-23 degrees C) and near neutral pH (7.2-7.5). C*T(99.9) values for chlorine increased >20-fold and for chloramine increased 2.6-fold when cells were grown in 100-fold diluted nutrient broth (2NB) solutions (final TOC of 35-40 mg/L). Background levels of Mg: 6.75 x 10(-2) mM and Fe: 3.58 x 10(-5) mM or high levels of FeCl3 (0.01 mM) and MgSO4 (1 mM) during growth resulted in the highest resistances to chlorine with C*T(99.9) values of 13.06 (+/-0.91) and 13.78 (+/-1.97) mg-min/L, respectively. Addition of low levels of FeCl3 (0.001 mM) and MgSO4 (0.1 mM) to K. pneumoniae cultures during growth resulted in the lowest bacterial resistances to inactivation; C*T(99.9) values ranged from 0.28 (+/-0.06) to 1.88 (+/-0.53)mg-min/L in these cultures. Increase in growth temperature from 22.5 degrees C to 35 degrees C for unamended 2NB cultures resulted in a 42-fold decrease in C*T(99.9) values for chlorine. A similar change in temperature resulted in no significant change in C*T(99.9) values for chloramine. These results indicate that inactivation of K. pneumoniae cultures by chlorine was highly sensitive to changes in growth conditions unlike inactivation by chloramine.

  8. Activity of Imipenem against Klebsiella pneumoniae Biofilms In Vitro and In Vivo

    PubMed Central

    Chen, Ping; Seth, Akhil K.; Abercrombie, Johnathan J.; Mustoe, Thomas A.

    2014-01-01

    Encapsulated Klebsiella pneumoniae has emerged as one of the most clinically relevant and more frequently encountered opportunistic pathogens in combat wounds as the result of nosocomial infection. In this report, we show that imipenem displayed potent activity against established K. pneumoniae biofilms under both static and flow conditions in vitro. Using a rabbit ear model, we also demonstrated that imipenem was highly effective against preformed K. pneumoniae biofilms in wounds. PMID:24247132

  9. Concurrent Endophthalmitis and Orbital Cellulitis From Metastatic Klebsiella pneumonia Liver Abscess.

    PubMed

    Davies, Brett W; Fante, Robert G

    2016-01-01

    A 70-year-old Korean female with a history of Klebsiella pneumonia liver abscess was presented to the authors' service with signs of endophthalmitis and orbital cellulitis. Vitreous biopsy confirmed K. pneumonia as the causative organism. With prolonged intravenous antibiotics and steroids, orbital symptoms resolved, but visual acuity remained at light perception. This is the first case in the literature to report on endophthalmitis and orbital cellulitis from K. pneumonia liver abscess.

  10. Recent Research Examining Links Among Klebsiella pneumoniae from Food, Food Animals, and Human Extraintestinal Infections.

    PubMed

    Davis, Gregg S; Price, Lance B

    2016-06-01

    Klebsiella pneumoniae is a colonizer of livestock, a contaminant of retail meats and vegetables, and a cause of extraintestinal infections in humans. Antibiotic-resistant strains of K. pneumoniae are becoming increasingly prevalent among hospital and community-acquired infections. Antibiotics are used extensively in conventional food-animal production, where they select for antibiotic-resistant bacteria. Antibiotic-resistant K. pneumoniae has been isolated from livestock as well as from a variety of retail meats, seafood, and vegetables. Furthermore, recent phylogenetic analyses suggest close relationships between K. pneumoniae from humans and livestock. Therefore, it is essential that we quantify the contribution of foodborne K. pneumoniae to antibiotic-resistant human infections.

  11. Community-onset Klebsiella pneumoniae pneumonia in Taiwan: clinical features of the disease and associated microbiological characteristics of isolates from pneumonia and nasopharynx.

    PubMed

    Lin, Yi-Tsung; Wang, Yu-Ping; Wang, Fu-Der; Fung, Chang-Phone

    2015-01-01

    Klebsiella pneumoniae is an important cause of community-onset pneumonia in Asian countries and South Africa. We investigated the clinical characteristics of K. pneumoniae causing community-onset pneumonia, and the associated microbiological features between K. pneumoniae isolates from pneumonia and those from the nasopharynx in Taiwan. This study was conducted at the Taipei Veterans General Hospital during July, 2012 to February, 2014. The clinical characteristics in patients with community-onset K. pneumoniae pneumonia were analyzed. K. pneumoniae isolates from the nasopharynx of adults attending otorhinolaryngology outpatient clinics were collected to compare their microbiological features with those from pneumonia. Capsular genotypes, antimicrobial susceptibility, and multilocus sequence type (MLST) were determined among these strains. Ninety-one patients with community-onset K. pneumoniae pneumonia were enrolled. We found a high mortality (29.7%) among these patients. Capsular types K1, K2, K5, K20, K54, and K57 accounted for ∼70% of the K. pneumoniae isolates causing pneumonia, and ∼70% of all the K. pneumoniae strains isolated from the nasopharynx of patients in outpatient clinics. The MLST profiles further demonstrated the genetic relatedness between most pneumonia isolates and those from the nasopharynx. In conclusion, our results show that community-onset pneumonia caused by K. pneumoniae was associated with high mortality and could have a reservoir in the nasopharynx. To tackle this high-mortality disease, the distribution of capsular types in the nasopharynx might have implications for future vaccine development.

  12. Community-onset Klebsiella pneumoniae pneumonia in Taiwan: clinical features of the disease and associated microbiological characteristics of isolates from pneumonia and nasopharynx

    PubMed Central

    Lin, Yi-Tsung; Wang, Yu-Ping; Wang, Fu-Der; Fung, Chang-Phone

    2015-01-01

    Klebsiella pneumoniae is an important cause of community-onset pneumonia in Asian countries and South Africa. We investigated the clinical characteristics of K. pneumoniae causing community-onset pneumonia, and the associated microbiological features between K. pneumoniae isolates from pneumonia and those from the nasopharynx in Taiwan. This study was conducted at the Taipei Veterans General Hospital during July, 2012 to February, 2014. The clinical characteristics in patients with community-onset K. pneumoniae pneumonia were analyzed. K. pneumoniae isolates from the nasopharynx of adults attending otorhinolaryngology outpatient clinics were collected to compare their microbiological features with those from pneumonia. Capsular genotypes, antimicrobial susceptibility, and multilocus sequence type (MLST) were determined among these strains. Ninety-one patients with community-onset K. pneumoniae pneumonia were enrolled. We found a high mortality (29.7%) among these patients. Capsular types K1, K2, K5, K20, K54, and K57 accounted for ∼70% of the K. pneumoniae isolates causing pneumonia, and ∼70% of all the K. pneumoniae strains isolated from the nasopharynx of patients in outpatient clinics. The MLST profiles further demonstrated the genetic relatedness between most pneumonia isolates and those from the nasopharynx. In conclusion, our results show that community-onset pneumonia caused by K. pneumoniae was associated with high mortality and could have a reservoir in the nasopharynx. To tackle this high-mortality disease, the distribution of capsular types in the nasopharynx might have implications for future vaccine development. PMID:25741336

  13. Outbreak of Klebsiella pneumoniae carbapenemase-producing K pneumoniae: A systematic review.

    PubMed

    Campos, Anaelís C; Albiero, James; Ecker, Alessandra B; Kuroda, Cristina M; Meirelles, Lívia E F; Polato, Angelita; Tognim, Maria C B; Wingeter, Márcia A; Teixeira, Jorge J V

    2016-11-01

    First detected in the United States in 1996, Klebsiella pneumoniae carbapenemase (KPC) has spread internationally among gram-negative bacteria, especially K pneumoniae. These microorganisms can cause serious infections in hospitalized patients, and there are few therapeutic options, culminating in increased mortality. The objective of this study was to describe the occurrence of outbreaks that were caused by KPC-producing K pneumoniae, emphasizing the interventions that were implemented to contain the outbreaks. PubMed, Web of Knowledge, and Literatura Latino Americana em Ciências da Saúde databases were searched for articles that were published between 2001 and 2012 according to the recommendations of Preferred Reporting Items for Systematic Reviews and Meta-Analyses. Of the 586 studies identified, 13 were selected for the final sample. Most studies showed that the containment of KPC outbreaks is possible in hospital settings through several actions, particularly use of surveillance cultures and the establishment of contact precautions. The results show that limiting the cross-transmission of these and other KPC-producing bacteria is possible in a hospital setting. However, such isolates need to be detected early with the aid of culture surveillance and contained early using appropriate actions immediately to prevent an outbreak. Copyright © 2016 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

  14. Therapeutic activities of cefazolin, cefotaxime, and ceftazidime against experimentally induced Klebsiella pneumoniae pneumonia in rats.

    PubMed Central

    Bakker-Woudenberg, I A; van den Berg, J C; Michel, M F

    1982-01-01

    The efficacies of several dosage schedules of cefazolin, cefotaxime, and ceftazidime, started 12 or 36 h after infection, were examined in experimental pneumonia caused by Klebsiella pneumoniae in rats. The therapeutic activities of the cephalosporins were compared with the antibacterial activities in vitro and the serum concentration curves. The course of experimental pneumonia was rapid and characterized by tissue necrosis. Response to antimicrobial treatment was evaluated with respect to mortality and numbers of bacteria in lung (left lobe), blood, and pleural fluid. When antibiotic treatment was started early, i.e., 12 h after bacterial inoculation, cefotaxime and ceftazidime were equally effective and superior to cefazolin. Eleven doses of 10 mg of cefotaxime or ceftazidime per kg or 11 doses of 60 mg of cefazolin per kg were required to improve the survival rate. With a delay in administration to 36 h after inoculation, the efficacy of the cephalosporins decreased markedly. In the three dosages tested, cefazolin was ineffective. Survival improved with the administration of nine doses of 60 mg of cefotaxime per kg or nine doses of 10 mg of ceftazidime per kg. These results are not in accordance with the ratio of in vitro activities of cefotaxime and ceftazidime or the serum concentration curves. Images PMID:6297384

  15. Rapidly fatal bacteremic pneumonia caused by Klebsiella pneumoniae with K1 hypermucoviscosity phenotype in a previously healthy young man receiving levofloxacin treatment.

    PubMed

    Chuang, Tzu-Yi; Lin, Chou-Jui; Chi, Chun-Lin; Liu, An-Yu; Lee, Shih-Wei; Lin, T L; Wang, Jin-Town; Hsueh, Po-Ren

    2009-10-01

    Fatal bacteremic Klebsiella pneumoniae pneumonia is commonly encountered in alcoholic and diabetic patients. This report describes a previously healthy young man with rapidly fatal bacteremic pneumonia caused by K. pneumoniae serotype K1, complicated by septic shock and multiple organ dysfunction.

  16. Identification and Characterization of Imipenem-Resistant Klebsiella pneumoniae and Susceptible Klebsiella variicola Isolates Obtained from the Same Patient.

    PubMed

    Garza-Ramos, Ulises; Moreno-Dominguez, Stephania; Hernández-Castro, Rigoberto; Silva-Sanchez, Jesús; Barrios, Humberto; Reyna-Flores, Fernando; Sanchez-Perez, Alejandro; Carrillo-Casas, Erika M; Sanchez-León, María Carmen; Moncada-Barron, David

    2016-04-01

    Klebsiella variicola, a bacterium closely genetically related to Klebsiella pneumoniae, is commonly misidentified as K. pneumoniae by biochemical tests. To distinguish between the two bacteria, phylogenetic analysis of the rpoB gene and the identification of unique genes in both bacterial species by multiplex-polymerase chain reaction (PCR) provide the means to reliably identify and genotype K. variicola. In recent years, K. variicola has been described both as the cause of an intrahospital outbreak in a pediatric hospital, which resulted in sepsis in inpatients, and as a frequent cause of bloodstream infections. In the present study, K. pneumoniae and K. variicola were isolated from a unique patient displaying different antimicrobial susceptibility phenotypes and different genotypes of virulence determinants. Eight clinical isolates were obtained at different time intervals; all during a 5-month period. The isolates were identified as K. pneumoniae by an automated identification system. The clinical (biochemical test) and molecular (multiplex-PCR and rpoB gene) characterization identified imipenem resistance in the first six K. pneumoniae ST258 isolates, which encode the SHV-12 cephalosporinase and KPC-3 carbapenemase genes. The two last remaining isolates corresponded to susceptible K. variicola. The bacterial species showed a specific profile of virulence-associated determinants, specifically the fimA, fimH, and ecpRAB fimbrial-encoding genes identified only in K. pneumoniae isolates. However, the entb (enterobactin), mrkD (fimbrial adhesin), uge (epimerase), ureA (urease), and wabG (transferase) genes were shared between both bacterial species. Recent studies attribute a higher mortality rate to K. variicola than to K. pneumonia. This work highlights the identification of K. pneumoniae and the closely related K. variicola isolated from the same patient. The value of distinguishing between these two bacterial species is in their clinical significance, their

  17. Predatory Bacteria Attenuate Klebsiella pneumoniae Burden in Rat Lungs.

    PubMed

    Shatzkes, Kenneth; Singleton, Eric; Tang, Chi; Zuena, Michael; Shukla, Sean; Gupta, Shilpi; Dharani, Sonal; Onyile, Onoyom; Rinaggio, Joseph; Connell, Nancy D; Kadouri, Daniel E

    2016-11-08

    Bdellovibrio bacteriovorus and Micavibrio aeruginosavorus are predatory bacteria that naturally-and obligately-prey on other Gram-negative bacteria, and their use has been proposed as a potential new approach to control microbial infection. The ability of predatory bacteria to prey on Gram-negative human pathogens in vitro is well documented; however, the in vivo safety and efficacy of predatory bacteria have yet to be fully assessed. In this study, we examined whether predatory bacteria can reduce bacterial burden in the lungs in an in vivo mammalian system. Initial safety studies were performed by intranasal inoculation of rats with predatory bacteria. No adverse effects or lung pathology were observed in rats exposed to high concentrations of predatory bacteria at up to 10 days postinoculation. Enzyme-linked immunosorbent assay (ELISA) of the immune response revealed a slight increase in inflammatory cytokine levels at 1 h postinoculation that was not sustained by 48 h. Additionally, dissemination experiments showed that predators were efficiently cleared from the host by 10 days postinoculation. To measure the ability of predatory bacteria to reduce microbial burden in vivo, we introduced sublethal concentrations of Klebsiella pneumoniae into the lungs of rats via intranasal inoculation and followed with multiple doses of predatory bacteria over 24 h. Predatory bacteria were able to reduce K. pneumoniae bacterial burden, on average, by more than 3.0 log10 in the lungs of most rats as measured by CFU plating. The work presented here provides further support for the idea of developing predatory bacteria as a novel biocontrol agent. A widely held notion is that antibiotics are the greatest medical advance of the last 50 years. However, the rise of multidrug-resistant (MDR) bacterial infections has become a global health crisis over the last decade. As we enter the postantibiotic era, it is crucial that we begin to develop new strategies to combat bacterial

  18. Regulation of anaerobic citrate metabolism in Klebsiella pneumoniae.

    PubMed

    Bott, M; Meyer, M; Dimroth, P

    1995-11-01

    Three enzymes are specifically required for uptake and catabolism of citrate by Klebsiella pneumoniae under anaerobic conditions: a Na+ -dependent citrate carrier (CitS), citrate lyase (CitDEF), and the Na+ pump oxaloacetate decarboxylase (OadGAB). The corresponding genes are clustered on the chromosome, with the citCDEFG genes located upstream and divergent to the citS-oadGAB genes. We found that expression of citS from its native promoter in Escherichia coli requires the DNA region downstream of oadB. Nucleotide sequence analysis of this region revealed the presence of two adjacent genes, citA and citB. By sequence similarity, the predicted CitA and CitB proteins were identified as members of the two-component regulatory systems. The sensor kinase CitA contained, in the N-terminal half, two putative transmembrane helices which enclosed a presumably periplasmic domain of about 130 amino acids. The C-terminal half of the response regulator CitB harboured a helix-turn-helix motif typical of DNA-binding proteins. K. pneumoniae citB null mutants were unable to grow anaerobically with citrate as the sole carbon and energy source (Cit- phenotype). When cultivated anaerobically with citrate plus glycerol, all of the citrate-specific fermentation enzymes were synthesized in the wild type, but not in the citB mutants. This showed that citS, oadGAB and citDEF required the CitB protein for expression and therefore are part of a regulon. In the wild type, synthesis of CitS, oxaloacetate decarboxylase and citrate lyase was dependent on the presence of citrate, sodium ions and a low oxygen tension. In a citA null mutant which expressed citB constitutively at high levels, none of these signals was required for the formation of the citrate fermentation enzymes. This result suggested that citrate, Na+, and oxygen exerted their regulatory effects via the CitA/CitB system. In the presence of these signals, the citAB gene products induced their own synthesis. The positive

  19. Klebsiella pneumoniae carbapenemases in Enterobacteriaceae: history, evolution, and microbiology concerns.

    PubMed

    Rapp, Robert P; Urban, Carl

    2012-05-01

    Since the discovery of penicillin 80 years ago, gram-negative bacteria have become proficient at evading the lethal activity of β-lactam antibiotics, principally through the production of β-lactamases. The rapid emergence of penicillinases in both gram-positive and gram-negative bacteria led to the development of cephalosporin β-lactam antibiotics, but production of plasmid-mediated extended-spectrum cephalosporinases (or extended-spectrum β-lactamases) and AmpC enzymes resulted in resistance to this drug class. Because carbapenems were the only β-lactam agents active against such extended-spectrum β-lactamase-producing strains, appropriate and inappropriate use soon resulted in Enterobacteriaceae resistance. As a result, two distinct types of carbapenemases-the metallo-β-lactamases and Klebsiella pneumoniae carbapenemases (KPCs)-were soon identified. The KPCs comprise 10 variants that differ from one another by one to three amino acid substitutions (KPC-2 to KPC-11). The KPC-producing Enterobacteriaceae are not only multidrug resistant but are also difficult to detect routinely in the clinical microbiology laboratory. Tigecycline, polymyxins (colistin and polymyxin B), and aminoglycosides are possible candidate therapies for infections caused by KPC-producing organisms, although well-conducted clinical trials are required to fully define their roles in patient management. The shortage of new antimicrobial agents on the immediate horizon suggests that enhanced adherence with infection prevention procedures and antimicrobial stewardship programs are needed to curb patient-to-patient transmission and to reduce the selection of multidrug-resistant bacteria.

  20. CXCR1/CXCR2 antagonism is effective in pulmonary defense against Klebsiella pneumoniae infection.

    PubMed

    Wei, Jing; Peng, Jing; Wang, Bing; Qu, Hong; Wang, Shiyi; Faisal, Aziz; Cheng, Jia-Wei; Gordon, John R; Li, Fang

    2013-01-01

    Klebsiella pneumoniae-associated pathology is largely mediated by neutrophilic inflammation. In this study, we administered Klebsiella pneumoniae to experimental guinea pig groups and the ELR-CXC chemokine antagonist CXCL8(3-72), ceftazidime, and dexamethasone to different groups, respectively. After 24 h, we assessed the animal's pulmonary inflammatory levels, including gross histopathology, airway neutrophilia, lung myeloperoxidase levels, expressions of CXCL8 and TNF, and airway bacterial loads. Compared with ceftazidime and dexamethasone treatments, the administration of the ELR-CXC chemokine antagonist CXCL8(3-72) alone was more effective than other methods, although it did not markedly attenuate the bacterial load. These results suggest new methods for the treatment of Klebsiella pneumoniae pathology.

  1. Destruction of single-species biofilms of Escherichia coli or Klebsiella pneumoniae subsp. pneumoniae by dextranase, lactoferrin, and lysozyme

    USDA-ARS?s Scientific Manuscript database

    The activity of dextranase, lactoferrin, lysozyme, and nisin against biofilms composed of either Klebsiella pneumonia or Escherichia coli was examined using the MBEC Assay™. Mature biofilms were treated and then sonicated to remove the adherent biofilm. This material was quantified using a lumines...

  2. Longitudinal Characterization of Acinetobacter baumannii-calcoaceticus Complex, Klebsiella pneumoniae, and Methicillin-Resistant Staphylococcus aureus Colonizing and Infecting Combat Casualties

    DTIC Science & Technology

    2012-01-01

    Brief report Longitudinal characterization of Acinetobacter baumannii-calcoaceticus complex, Klebsiella pneumoniae , and methicillin-resistant...resistant Acinetobacter baumannii-calcoaceticus complex Klebsiella pneumoniae Methicillin-resistant Staphylococcus aureus MRSA Drug-resistant...Acinetobacter baumannii-calcoaceticus complex, Klebsiella pneumoniae , and methicillin- resistant Staphylococcus aureus colonize and infect combat casualties

  3. Occurrence of Diverse Antimicrobial Resistance Determinants in Genetically Unrelated Biocide Tolerant Klebsiella pneumoniae

    PubMed Central

    Mondal, Amitabha; Venkataramaiah, Manjunath; Rajamohan, Govindan; Srinivasan, Vijaya Bharathi

    2016-01-01

    Nosocomial infections due to Klebsiella pneumoniae is a significant problem in health care settings worldwide. In this study, we examined the antimicrobial susceptibility, genetic profiles and mechanisms of antibiotic resistance in K. pneumoniae isolates of Indian origin. To our knowledge this is the first report demonstrating the high prevalence of β-lactamases, aminoglycoside modifying enzymes, quinolone resistance genes besides demonstrating the involvement of active efflux in K. pneumoniae Indian isolates. This study has enabled us to correlate the phenotypic and genotypic characteristics in K. pneumoniae, providing an important base for continued monitoring and epidemiological studies of this emerging nosocomial pathogen in Indian hospitals. PMID:27870879

  4. Dissemination of the KPC-2 carbapenemase in non-Klebsiella pneumoniae enterobacterial isolates from Colombia.

    PubMed

    Cuzon, Gaelle; Naas, Thierry; Correa, Adriana; Quinn, John P; Villegas, Maria-Virginia; Nordmann, Patrice

    2013-07-01

    Klebsiella pneumoniae carbapenemase (KPC)-type enzymes have largely disseminated worldwide among K. pneumoniae isolates. In this study, 11 non-K. pneumoniae KPC-producing enterobacterial isolates from four hospitals located in different Colombian cities were genetically investigated. All isolates were multidrug-resistant and harboured the bla(KPC-2) gene along with several other acquired β-lactamase genes. The bla(KPC-2) gene was associated with transposon Tn4401b inserted in different loci of plasmids varying in size and replicon type. The presence of KPC-2 in different enterobacterial species from different cities within Colombia underlines the spread of KPC beyond K. pneumoniae.

  5. Modeling Klebsiella pneumoniae pathogenesis by infection of the wax moth Galleria mellonella.

    PubMed

    Insua, José Luis; Llobet, Enrique; Moranta, David; Pérez-Gutiérrez, Camino; Tomás, Anna; Garmendia, Junkal; Bengoechea, José A

    2013-10-01

    The implementation of infection models that approximate human disease is essential for understanding pathogenesis at the molecular level and for testing new therapies before they are entered into clinical stages. Insects are increasingly being used as surrogate hosts because they share, with mammals, essential aspects of the innate immune response to infections. We examined whether the larva of the wax moth Galleria mellonella could be used as a host model to conceptually approximate Klebsiella pneumoniae-triggered pneumonia. We report that the G. mellonella model is capable of distinguishing between pathogenic and nonpathogenic Klebsiella strains. Moreover, K. pneumoniae infection of G. mellonella models some of the known features of Klebsiella-induced pneumonia, i.e., cell death associated with bacterial replication, avoidance of phagocytosis by phagocytes, and the attenuation of host defense responses, chiefly the production of antimicrobial factors. Similar to the case for the mouse pneumonia model, activation of innate responses improved G. mellonella survival against subsequent Klebsiella challenge. Virulence factors necessary in the mouse pneumonia model were also implicated in the Galleria model. We found that mutants lacking capsule polysaccharide, lipid A decorations, or the outer membrane proteins OmpA and OmpK36 were attenuated in Galleria. All mutants activated G. mellonella defensive responses. The Galleria model also allowed us to monitor Klebsiella gene expression. The expression levels of cps and the loci implicated in lipid A remodeling peaked during the first hours postinfection, in a PhoPQ- and PmrAB-governed process. Taken together, these results support the utility of G. mellonella as a surrogate host for assessing infections with K. pneumoniae.

  6. Modeling Klebsiella pneumoniae Pathogenesis by Infection of the Wax Moth Galleria mellonella

    PubMed Central

    Insua, José Luis; Llobet, Enrique; Moranta, David; Pérez-Gutiérrez, Camino; Tomás, Anna; Garmendia, Junkal

    2013-01-01

    The implementation of infection models that approximate human disease is essential for understanding pathogenesis at the molecular level and for testing new therapies before they are entered into clinical stages. Insects are increasingly being used as surrogate hosts because they share, with mammals, essential aspects of the innate immune response to infections. We examined whether the larva of the wax moth Galleria mellonella could be used as a host model to conceptually approximate Klebsiella pneumoniae-triggered pneumonia. We report that the G. mellonella model is capable of distinguishing between pathogenic and nonpathogenic Klebsiella strains. Moreover, K. pneumoniae infection of G. mellonella models some of the known features of Klebsiella-induced pneumonia, i.e., cell death associated with bacterial replication, avoidance of phagocytosis by phagocytes, and the attenuation of host defense responses, chiefly the production of antimicrobial factors. Similar to the case for the mouse pneumonia model, activation of innate responses improved G. mellonella survival against subsequent Klebsiella challenge. Virulence factors necessary in the mouse pneumonia model were also implicated in the Galleria model. We found that mutants lacking capsule polysaccharide, lipid A decorations, or the outer membrane proteins OmpA and OmpK36 were attenuated in Galleria. All mutants activated G. mellonella defensive responses. The Galleria model also allowed us to monitor Klebsiella gene expression. The expression levels of cps and the loci implicated in lipid A remodeling peaked during the first hours postinfection, in a PhoPQ- and PmrAB-governed process. Taken together, these results support the utility of G. mellonella as a surrogate host for assessing infections with K. pneumoniae. PMID:23836821

  7. Distinct Contributions of Neutrophils and CCR2+ Monocytes to Pulmonary Clearance of Different Klebsiella pneumoniae Strains.

    PubMed

    Xiong, Huizhong; Carter, Rebecca A; Leiner, Ingrid M; Tang, Yi-Wei; Chen, Liang; Kreiswirth, Barry N; Pamer, Eric G

    2015-09-01

    Klebsiella pneumoniae is a common respiratory pathogen, with some strains having developed broad resistance to clinically available antibiotics. Humans can become infected with many different K. pneumoniae strains that vary in genetic background, antibiotic susceptibility, capsule composition, and mucoid phenotype. Genome comparisons have revealed differences between K. pneumoniae strains, but the impact of genomic variability on immune-mediated clearance of pneumonia remains unclear. Experimental studies of pneumonia in mice have used the rodent-adapted 43816 strain of K. pneumoniae and demonstrated that neutrophils are essential for optimal host defense. It remains unclear, however, whether CCR2(+) monocytes contribute to K. pneumoniae clearance from the lung. We selectively depleted neutrophils, CCR2(+) monocytes, or both from immunocompetent mice and determined susceptibility to infection by the 43816 strain and 4 newly isolated clinical K. pneumoniae strains. The clinical K. pneumoniae strains, including one carbapenem-resistant ST258 strain, are less virulent than 43816. Optimal clearance of each of the 5 strains required either neutrophils or CCR2(+) monocytes. Selective neutrophil depletion markedly worsened infection with K. pneumoniae strain 43816 and three clinical isolates but did not increase susceptibility of mice to infection with the carbapenem-resistant K. pneumoniae ST258 strain. Depletion of CCR2(+) monocytes delayed recovery from infection with each of the 5 K. pneumoniae strains, revealing a contribution of these cells to bacterial clearance from the lung. Our findings demonstrate strain-dependent variation in the contributions of neutrophils and CCR2(+) monocytes to clearance of K. pneumoniae pulmonary infection.

  8. Antimicrobial resistance of Klebsiella pneumoniae stool isolates circulating in Kenya.

    PubMed

    Taitt, Chris Rowe; Leski, Tomasz A; Erwin, Daniel P; Odundo, Elizabeth A; Kipkemoi, Nancy C; Ndonye, Janet N; Kirera, Ronald K; Ombogo, Abigael N; Walson, Judd L; Pavlinac, Patricia B; Hulseberg, Christine; Vora, Gary J

    2017-01-01

    We sought to determine the genetic and phenotypic antimicrobial resistance (AMR) profiles of commensal Klebsiella spp. circulating in Kenya by testing human stool isolates of 87 K. pneumoniae and three K. oxytoca collected at eight locations. Over one-third of the isolates were resistant to ≥3 categories of antimicrobials and were considered multidrug-resistant (MDR). We then compared the resistance phenotype to the presence/absence of 238 AMR genes determined by a broad-spectrum microarray and PCR. Forty-six genes/gene families were identified conferring resistance to β-lactams (ampC/blaDHA, blaCMY/LAT, blaLEN-1, blaOKP-A/OKP-B1, blaOXA-1-like family, blaOXY-1, blaSHV, blaTEM, blaCTX-M-1 and blaCTX-M-2 families), aminoglycosides (aac(3)-III, aac(6)-Ib, aad(A1/A2), aad(A4), aph(AI), aph3/str(A), aph6/str(B), and rmtB), macrolides (mac(A), mac(B), mph(A)/mph(K)), tetracyclines (tet(A), tet(B), tet(D), tet(G)), ansamycins (arr), phenicols (catA1/cat4, floR, cmlA, cmr), fluoroquinolones (qnrS), quaternary amines (qacEΔ1), streptothricin (sat2), sulfonamides (sul1, sul2, sul3), and diaminopyrimidines (dfrA1, dfrA5, dfrA7, dfrA8, dfrA12, dfrA13/21/22/23 family, dfrA14, dfrA15, dfrA16, dfrA17). This is the first profile of genes conferring resistance to multiple categories of antimicrobial agents in western and central Kenya. The large number and wide variety of resistance genes detected suggest the presence of significant selective pressure. The presence of five or more resistance determinants in almost two-thirds of the isolates points to the need for more effective, targeted public health policies and infection control/prevention measures.

  9. Immunogenic properties of Klebsiella pneumoniae type 2 capsular polysaccharide.

    PubMed Central

    Robert, A; Jouin, H; Fournier, J M

    1986-01-01

    The immunoprotective activity of Klebsiella pneumoniae K2 cell surface preparations and purified capsular polysaccharide was tested in mice. The 50% protective dose (PD50), expressed as capsular polysaccharide content, was 2 ng for cell surface preparations and 50 ng for purified capsular polysaccharide. Both preparations lost their immunoprotective activity after alkali treatment. Immune sera were raised in rabbits immunized with cell surface preparations. The precipitating and hemagglutinating capacity of these antisera was tested against either purified capsular polysaccharide or alkali-treated capsular polysaccharide. No difference was observed between the reactivity of the antisera against each antigen. The protective activity of these sera was tested on mice in passive transfer experiments, before and after absorption with either purified capsular polysaccharide or alkali-treated capsular polysaccharide. The sera lost their protective activity after absorption with purified capsular polysaccharide and after absorption with alkali-treated capsular polysaccharide. These experiments show that the difference in immunoprotective activity of cell surface preparations, purified capsular polysaccharide, and alkali-treated capsular polysaccharide is not due to a difference in their antigenic determinants. Cell surface preparations and purified capsular polysaccharide were fractionated by gel filtration on Sepharose 4B and by ultracentrifugation on cesium chloride density gradients. Three forms of capsular polysaccharide have been characterized. (i) A form of capsular polysaccharide with a very high protective activity (PD50 = 2 ng) that copurified with protein and lipopolysaccharide and was characterized by a low coefficient of distribution (Kd = 0.20) and a low density (1.5 to 1.6 g/cm3). (ii) A form of capsular polysaccharide with an intermediate protective activity (PD50 = 50 ng), contamined by less than 3% protein and 1% lipopolysaccharide, with a Kd of 0.35, and

  10. Antimicrobial resistance of Klebsiella pneumoniae stool isolates circulating in Kenya

    PubMed Central

    Leski, Tomasz A.; Erwin, Daniel P.; Odundo, Elizabeth A.; Kipkemoi, Nancy C.; Ndonye, Janet N.; Kirera, Ronald K.; Ombogo, Abigael N.; Walson, Judd L.; Pavlinac, Patricia B.; Hulseberg, Christine; Vora, Gary J.

    2017-01-01

    We sought to determine the genetic and phenotypic antimicrobial resistance (AMR) profiles of commensal Klebsiella spp. circulating in Kenya by testing human stool isolates of 87 K. pneumoniae and three K. oxytoca collected at eight locations. Over one-third of the isolates were resistant to ≥3 categories of antimicrobials and were considered multidrug-resistant (MDR). We then compared the resistance phenotype to the presence/absence of 238 AMR genes determined by a broad-spectrum microarray and PCR. Forty-six genes/gene families were identified conferring resistance to β-lactams (ampC/blaDHA, blaCMY/LAT, blaLEN-1, blaOKP-A/OKP-B1, blaOXA-1-like family, blaOXY-1, blaSHV, blaTEM, blaCTX-M-1 and blaCTX-M-2 families), aminoglycosides (aac(3)-III, aac(6)-Ib, aad(A1/A2), aad(A4), aph(AI), aph3/str(A), aph6/str(B), and rmtB), macrolides (mac(A), mac(B), mph(A)/mph(K)), tetracyclines (tet(A), tet(B), tet(D), tet(G)), ansamycins (arr), phenicols (catA1/cat4, floR, cmlA, cmr), fluoroquinolones (qnrS), quaternary amines (qacEΔ1), streptothricin (sat2), sulfonamides (sul1, sul2, sul3), and diaminopyrimidines (dfrA1, dfrA5, dfrA7, dfrA8, dfrA12, dfrA13/21/22/23 family, dfrA14, dfrA15, dfrA16, dfrA17). This is the first profile of genes conferring resistance to multiple categories of antimicrobial agents in western and central Kenya. The large number and wide variety of resistance genes detected suggest the presence of significant selective pressure. The presence of five or more resistance determinants in almost two-thirds of the isolates points to the need for more effective, targeted public health policies and infection control/prevention measures. PMID:28575064

  11. In Vivo Emergence of Tigecycline Resistance in Multidrug-Resistant Klebsiella pneumoniae and Escherichia coli

    PubMed Central

    Spanu, Teresa; De Angelis, Giulia; Cipriani, Michela; Pedruzzi, Barbara; D'Inzeo, Tiziana; Cataldo, Maria Adriana; Sganga, Gabriele

    2012-01-01

    Although resistance to tigecycline has been reported in surveillance studies, very few reports have described the emergence of resistance in vivo. We report two cases of patients with infections due to SHV-12-producing Klebsiella pneumoniae and K. pneumoniae carbapenemase-3 (KPC-3)-producing Escherichia coli, which developed tigecycline resistance in vivo after treatment. The reported limited experience underlines the risk of occurrence of a tigecycline MIC increase under treatment pressure. PMID:22644031

  12. First Report of Ceftazidime-Avibactam Resistance in a KPC-3-Expressing Klebsiella pneumoniae Isolate

    PubMed Central

    Yang, Shangxin; Hemarajata, Peera; Ward, Kevin W.; Miller, Shelley A.; Gregson, Aric

    2015-01-01

    Ceftazidime-avibactam is the first antimicrobial approved by the U.S. FDA for the treatment of carbapenem-resistant Enterobacteriaceae. Avibactam, a non-β-lactam β-lactamase inhibitor, inactivates class A serine carbapenemases, including Klebsiella pneumoniae carbapenemase (KPC). We report a KPC-producing K. pneumoniae isolate resistant to ceftazidime-avibactam (MIC, 32/4 μg/ml) from a patient with no prior treatment with ceftazidime-avibactam. PMID:26195508

  13. A case of lobar pneumonia and sepsis with death caused by invasive Klebsiella rhinoscleromatis infection.

    PubMed

    Kumade, Eri; Furusyo, Norihiro; Takeshima, Norito; Kishihara, Yasuhiro; Mitsumoto-Kaseida, Fujiko; Etoh, Yoshitaka; Murata, Masayuki; Hayashi, Jun

    2016-10-01

    Klebsiella pneumoniae often causes pneumonia and other infections in heavy drinkers and patients with diabetes. Pneumonia caused by Klebsiella rhinoscleromatis, a subspecies of K. pneumoniae, has not been previously reported. We report a case of pneumonia caused by K. rhinoscleromatis. A 68-year-old man with type 2 diabetes visited our department complaining fever and fatigue for 10 days and cough and bloody sputum for two days. His Japan Coma Scale score was I-1, body temperature 38.3 °C, blood pressure 85/51 mmHg, pulse 135 bpm, and peripheral capillary oxygen saturation level 92% (room air). He had no abnormal breathing sounds. His white blood cell count had decreased to 2600/μL, and his C-reactive protein level was high, at 35.9 mg/dL. Chest computed tomography revealed lobar pneumonia in the right upper lobe and pneumonia in the left upper division. Klebsiella was suspected based on the result of a sputum smear examination. He was diagnosed with septic shock due to pneumonia and was immediately admitted. Intravenous antibacterial (levofloxacin) treatment was initiated, however, he died 13 h after presenting at the hospital. Subsequently, K. rhinoscleromatis was detected in sputum and blood culture. Additional testing determined the bacteria to be a highly pathogenic hypermucoviscosity phenotype and the cause of the fatal lobar pneumonia. Although cases of rhinoscleroma and bacteremia caused by K. rhinoscleromatis infection have been reported, this is the first report of a case with sepsis caused by fulminant pneumonia. Copyright © 2016 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  14. Tumor necrosis factor mediates lung antibacterial host defense in murine Klebsiella pneumonia.

    PubMed Central

    Laichalk, L L; Kunkel, S L; Strieter, R M; Danforth, J M; Bailie, M B; Standiford, T J

    1996-01-01

    Tumor necrosis factor (TNF) is a proinflammatory cytokine which has recently been shown to have beneficial effects in the setting of acquired host immunity. However, the role of TNF in innate immune responses, as in the setting of bacterial pneumonia, has been incompletely characterized. To determine the role of TNF in gram-negative bacterial pneumonia, CBA/J mice were challenged with 10(2) CFU of Klebsiella pneumoniae intratracheally, resulting in the time-dependent expression of TNF MRNA and protein within the lung. Passive immunization of animals with a soluble TNF receptor-immunoglobulin (Ig) construct (sTNFR:Fc) intraperitoneally 2 h prior to K. pneumoniae inoculation resulted in a significant reduction in bronchoalveolar lavage neutrophils, but not macrophages, at 48 h, as compared with animals receiving control IgG1. Furthermore, treatment with sTNFR:Fc resulted in 19.6- and 13.5-fold increases in K. pneumoniae CFU in lung homogenates and plasma, respectively, as compared with animals receiving control IgG1. Finally, treatment of Klebsiella-infected mice with sTNFR:Fc markedly decreased both short- and long-term survival of these animals. In conclusion, our studies indicate that endogenous TNF is a critical component of antibacterial host defense in murine Klebsiella pneumonia. PMID:8945568

  15. Bacteremic community-acquired pneumonia due to Klebsiella pneumoniae: clinical and microbiological characteristics in Taiwan, 2001-2008.

    PubMed

    Lin, Yi-Tsung; Jeng, Yuan-Yu; Chen, Te-Li; Fung, Chang-Phone

    2010-10-25

    Klebsiella pneumoniae is the major cause of community-acquired pyogenic infections in Taiwan. This retrospective study evaluated the clinical and microbiological characteristics of bacteremic community-acquired pneumonia due to K. pneumoniae in Taiwanese adults. The clinical characteristics of bacteremic community-acquired pneumonia (CAP) in adults due to K. pneumoniae were compared to those of adults with bacteremic CAP due to Streptococcus pneumoniae at a tertiary medical center in Taiwan from 2001-2008. Risk factors for mortality of bacteremic CAP due to K. pneumoniae were analyzed. All clinical isolates of K. pneumoniae were examined for capsular serotypes, hypermucoviscosity phenotype, aerobactin and rmpA gene. K. pneumoniae was the dominant cause of bacteremic CAP and was associated with a more fulminant course and a worse prognosis than bacteremic CAP due to Streptococcus pneumoniae. Initial presentation with septic shock and respiratory failure were independent risk factors for both early and total mortality. Serotype K1 and K2 comprised around half of all isolates. There were no significant differences in the clinical characteristics of patients with bacteremic CAP due to K1/K2 and non-K1/K2 isolates. Hypermucoviscosity phenotype as well as the aerobactin and rmpA genes were highly prevalent in the K. pneumoniae isolates. K. pneumoniae continued to be the dominant cause of bacteremic CAP in Taiwanese adults during 2001-2008. Initial presentation with septic shock and respiratory failure were independent risk factors for both early and total mortality from K. pneumoniae bacteremic CAP. Serotypes K1/K2 comprised around half of all isolates, but did not predispose patients to a poor clinical outcome. Physicians should be aware of the poor prognosis of any patient with bacteremic K. pneumoniae CAP and monitor these patients more closely.

  16. Klebsiella pneumoniae infection on a rehabilitation unit: comparison of epidemiologic typing methods.

    PubMed

    Thompson, W; Romance, L; Bialkowska-Hobrazanska, H; Rennie, R P; Ashton, F; Nicolle, L E

    1993-04-01

    To identify factors associated with an increased occurrence of Klebsiella pneumoniae isolation in urine cultures and infected wounds on a rehabilitation unit and to compare typing methods for K pneumoniae isolates. Retrospective review of laboratory reports and patient records with case-control study. Analysis of K pneumoniae isolates using capsular serotyping, enzyme electrophoretic typing, ribotyping, and DNA typing. 48-bed rehabilitation unit in an 1,100-bed tertiary care teaching hospital in Winnipeg, Manitoba. In 1988, 20 (19%) of 106 patients admitted to the rehabilitation unit had K pneumoniae isolated from urine or wound, and in 1989 31 (28%) of 111 patients had Klebsiella isolated. Review of ward practices revealed appropriate written policies but evidence of failure in execution leading to multiple opportunities for transmission among patients. Substantial environmental contamination was not identified, although a common urine graduate may have contributed to some transmission. Individuals with K pneumoniae isolated had a significantly longer duration of stay. Many of these were spinal cord-injured patients and were maintained on intermittent catheterization. One outbreak strain was identified in epidemiologic typing. Other strains were generally identified in individuals with non-nosocomial acquisition of infection. Comparison of epidemiologic typing methods suggests ribotyping may be the optimal method for typing K pneumoniae strains. K pneumoniae was acquired frequently by spinal cord-injured patients with extended admissions, re-emphasizing the importance of both patients and staff following appropriate infection control practices on rehabilitation wards. Ribotyping was the optimal method for typing K pneumoniae isolates.

  17. Transgenic tissue-type plasminogen activator expression improves host defense during Klebsiella pneumonia.

    PubMed

    Renckens, R; Roelofs, J J T H; Stegenga, M E; Florquin, S; Levi, M; Carmeliet, P; Van't Veer, C; van der Poll, T

    2008-04-01

    Severe pneumonia is associated with a local inhibition of fibrinolysis in the lung as reflected by strongly reduced pulmonary plasminogen activator activity. To study the effect of elevation of local plasminogen activator activity during pneumonia caused by the common respiratory pathogen Klebsiella pneumoniae. Female C57Bl/6 mice were inoculated intranasally with a replication-defective adenoviral vector expressing human tissue-type plasminogen activator or a control vector 24 h before intranasal infection with K. pneumoniae. Mice infected with Klebsiella via the airways developed overt pneumonia, which was accompanied by a downregulation of pulmonary tissue-type plasminogen activator levels at protein and mRNA levels. Pulmonary overexpression of human tissue-type plasminogen activator resulted in increased fibrinolytic activity in the lungs during pneumonia, as indicated by higher D-dimer levels and reduced fibrin deposition. Interestingly, overexpression of tissue-type plasminogen activator markedly improved host defense against pneumonia: mice treated with the tissue-type plasminogen activator vector displayed less bacterial growth and dissemination, attenuated distant organ injury and a reduced mortality. These data demonstrate that local elevation of plasminogen activator activity in the lungs improves host defense against severe gram-negative pneumonia and sepsis.

  18. Production of 2,3-butylene glycol from whey by Klebsiella pneumoniae and Enterobacter aerogenes.

    PubMed

    Barrett, E L; Collins, E B; Hall, B J; Matoi, S H

    1983-12-01

    Production of 2,3-butylene glycol from whey with Klebsiella pneumoniae and Enterobacter aerogenes was studied. Sterilization of the whey was unnecessary. Acid whey required neutralization, but sweet whey did not. Butylene glycol production was most efficient at 33 degrees C for Klebsiella pneumoniae and at 37 degrees C for Enterobacter aerogenes. Aeration significantly improved yields. Klebsiella pneumoniae produced more butylene glycol than did Enterobacter aerogenes in unsupplemented whey. The addition of 50 mM sodium acetate to whey increased the production of butylene glycol and acetoin by Enterobacter aerogenes; it also increased the production of glycol by Klebsiella pneumoniae, but the increase in this case was offset by a decrease of production of acetoin. Maximal yields of the glycol plus acetoin in whey were obtained in 48 to 64 h, but Enterobacter aerogenes required about 160 h for complete utilization of the lactose. Highest yields were about .3 M butylene glycol plus acetoin, which corresponds to the production of about 10 kg of glycol from 380 liters of whey.

  19. Phenotypic and Molecular Characterization of Multidrug Resistant Klebsiella pneumoniae Isolated from a University Teaching Hospital, China

    PubMed Central

    Liu, Helu; Lü, Dongyue; Liang, Hong; Dou, Yuhong

    2014-01-01

    The multidrug-resistant rate of Klebsiella pneumoniae has risen rapidly worldwide. To better understand the multidrug resistance situation and molecular characterization of Klebsiella pneumoniae, a total of 153 Klebsiella pneumoniae isolates were collected, and drug susceptibility test was performed to detect its susceptibility patterns to 13 kinds of antibiotics. Phenotypic tests for carbapenemases ESBLs and AmpC enzyme-producing strains were performed to detect the resistance phenotype of the isolates. Then PCR amplification and sequencing analysis were performed for the drug resistance determinants. The results showed that 63 strains harbored blaCTX-M gene, and 14 strains harbored blaDHA gene. Moreover, there were 5 strains carrying blaKPC gene, among which 4 strains carried blaCTX-M, blaDHA and blaKPC genes, and these 4 strains were also resistant to imipenem. Our data indicated that drug-resistant Klebsiella pneumoniae were highly prevalent in the hospital. Thus it is warranted that surveillance of epidemiology of those resistant isolates should be a cause for concern, and appropriate drugs should be chosen. PMID:24740167

  20. Novel VIM metallo-beta-lactamase variant, VIM-24, from a Klebsiella pneumoniae isolate from Colombia.

    PubMed

    Montealegre, Maria Camila; Correa, Adriana; Briceño, David F; Rosas, Natalia C; De La Cadena, Elsa; Ruiz, Sory J; Mojica, Maria F; Camargo, Ruben Dario; Zuluaga, Ivan; Marin, Adriana; Quinn, John P; Villegas, Maria Virginia

    2011-05-01

    We report the emergence of a novel VIM variant (VIM-24) in a Klebsiella pneumoniae isolate in Colombia. The isolate displays MICs for carbapenems below the resistance breakpoints, posing a real challenge for its detection. The blaVIM-24 gene was located within a class 1 integron carried on a large plasmid. Further studies are needed to clarify its epidemiological and clinical impact.

  1. High-level resistance to cefotaxime and ceftazidime in Klebsiella pneumoniae isolates from Cleveland, Ohio.

    PubMed Central

    Thomson, K S; Sanders, C C; Washington, J A

    1991-01-01

    Two isolates of Klebsiella pneumoniae possessing both TEM-1 and SHV-2 beta-lactamases were isolated from patients at the Cleveland Clinic in 1988. The beta-lactamases were discriminated and identified by using substrate hydrolysis data and an isoelectric focusing procedure in which the gel was overlaid with beta-lactamase inhibitors. Images PMID:1854155

  2. Immunoproteomic to Analysis the Pathogenicity Factors in Leukopenia Caused by Klebsiella Pneumonia Bacteremia

    PubMed Central

    Liu, Haiyan; Cheng, Zhongle; Song, Wen; Wu, Wenyong; Zhou, Zheng

    2014-01-01

    Incidences of leukopenia caused by bacteremia have increased significantly and it is associated with prolonged hospital stay and increased cost. Immunoproteomic is a promising method to identify pathogenicity factors of different diseases. In the present study, we used immunoproteomic to analysis the pathogenicity factors in leukopenia caused by Klebsiella Pneumonia bacteremia. Approximately 40 protein spots localized in the 4 to 7 pI range were detected on two-dimensional electrophoresis gels, and 6 differentially expressed protein spots between 10 and 170 kDa were identified. Pathogenicity factors including S-adenosylmethionine synthetase, pyruvate dehydrogenase, glutathione synthetase, UDP-galactose-4-epimerase, acetate kinase A and elongation factor tu (EF-Tu). In validation of the pathogenicity factor, we used western blotting to show that Klebsiella pneumonia had higher (EF-Tu) expression when they accompanied by leukopenia rather than leukocytosis. Thus, we report 6 pathogenicity factors of leukopenia caused by Klebsiella pneumonia bacteremia, including 5 housekeeping enzymes and EF-Tu. We suggest EF-Tu could be a potential pathogenicity factor for leukopenia caused by Klebsiella pneumonia. PMID:25330314

  3. Phenotypic and molecular characterization of multidrug resistant Klebsiella pneumoniae isolated from a university teaching hospital, China.

    PubMed

    Du, Jikun; Li, Peipei; Liu, Helu; Lü, Dongyue; Liang, Hong; Dou, Yuhong

    2014-01-01

    The multidrug-resistant rate of Klebsiella pneumoniae has risen rapidly worldwide. To better understand the multidrug resistance situation and molecular characterization of Klebsiella pneumoniae, a total of 153 Klebsiella pneumoniae isolates were collected, and drug susceptibility test was performed to detect its susceptibility patterns to 13 kinds of antibiotics. Phenotypic tests for carbapenemases ESBLs and AmpC enzyme-producing strains were performed to detect the resistance phenotype of the isolates. Then PCR amplification and sequencing analysis were performed for the drug resistance determinants. The results showed that 63 strains harbored bla CTX-M gene, and 14 strains harbored bla DHA gene. Moreover, there were 5 strains carrying bla KPC gene, among which 4 strains carried bla CTX-M, bla DHA and bla KPC genes, and these 4 strains were also resistant to imipenem. Our data indicated that drug-resistant Klebsiella pneumoniae were highly prevalent in the hospital. Thus it is warranted that surveillance of epidemiology of those resistant isolates should be a cause for concern, and appropriate drugs should be chosen.

  4. Impact of Extended Spectrum Beta-Lactamase Producing Klebsiella pneumoniae Infections in Severely Burned Patients

    DTIC Science & Technology

    2010-09-01

    there is no copyright to be transferred. Received February 6, 2010; Revised March 31, 2010; Accepted March 31, 2010. From Walter Reed Army Institute... Fung CP, et al. Variety of TEM-, SHV-, and CTX-M-type beta-lactamases present in recent clinical isolates of Escherichia coli, Klebsiella pneumoniae

  5. Label-Free Proteome Profiling of Carbapenem-Resistant Klebsiella pneumoniae LC-MS/MS

    DTIC Science & Technology

    2016-12-12

    Title Carbapenem- resistant Klebsiella pneumoniae LC-MS/MS Description A comprehensive analysis of proteome changes in CRKP cells in response to...the sub-MIC doses of antibiotics was performed by using label-free quantitative mass spectrometry. Sample Processing Protocol The pellets were

  6. Klebsiella pneumoniae Strains Producing Extended-Spectrum β-Lactamases in Spain: Microbiological and Clinical Features▿

    PubMed Central

    de Alegría, C. Ruiz; Rodríguez-Baño, J.; Cano, M. E.; Hernández-Bello, J. R.; Calvo, J.; Román, E.; Díaz, M. A.; Pascual, A.; Martínez-Martínez, L.

    2011-01-01

    Extended-spectrum β-lactamases (ESBL) of the CTX-M, SHV, and TEM families were recognized in 76 (67%), 31 (27%), and 6 (5%) isolates, respectively, among 162 ESBL-producing Klebsiella pneumoniae (ESBL-Kp) strains obtained in a multicenter study in Spain. Predisposing factors for ESBL-Kp acquisition included invasive procedures, mechanical ventilation, and previous antimicrobial use. PMID:21191059

  7. Klebsiella pneumoniae strains producing extended-spectrum beta-lactamases in Spain: microbiological and clinical features.

    PubMed

    Ruiz de Alegría, C; Rodríguez-Baño, J; Cano, M E; Hernández-Bello, J R; Calvo, J; Román, E; Díaz, M A; Pascual, A; Martínez-Martínez, L

    2011-03-01

    Extended-spectrum β-lactamases (ESBL) of the CTX-M, SHV, and TEM families were recognized in 76 (67%), 31 (27%), and 6 (5%) isolates, respectively, among 162 ESBL-producing Klebsiella pneumoniae (ESBL-Kp) strains obtained in a multicenter study in Spain. Predisposing factors for ESBL-Kp acquisition included invasive procedures, mechanical ventilation, and previous antimicrobial use.

  8. mcr-1 Colistin Resistance in ESBL-Producing Klebsiella pneumoniae, France.

    PubMed

    Caspar, Yvan; Maillet, Mylène; Pavese, Patricia; Francony, Gilles; Brion, Jean-Paul; Mallaret, Marie-Reine; Bonnet, Richard; Robin, Frédéric; Beyrouthy, Racha; Maurin, Max

    2017-05-01

    We report intestinal carriage of an extended-spectrum β-lactamase-producing Klebsiella pneumoniae strain with high-level resistance to colistin (MIC 24 mg/L) in a patient in France who had been hospitalized for fungal meningitis. The strain had the mcr-1 plasmid gene and an inactivated mgrB gene, which are associated with colistin resistance.

  9. Immunoproteomic to analysis the pathogenicity factors in leukopenia caused by Klebsiella pneumonia bacteremia.

    PubMed

    Liu, Haiyan; Cheng, Zhongle; Song, Wen; Wu, Wenyong; Zhou, Zheng

    2014-01-01

    Incidences of leukopenia caused by bacteremia have increased significantly and it is associated with prolonged hospital stay and increased cost. Immunoproteomic is a promising method to identify pathogenicity factors of different diseases. In the present study, we used immunoproteomic to analysis the pathogenicity factors in leukopenia caused by Klebsiella Pneumonia bacteremia. Approximately 40 protein spots localized in the 4 to 7 pI range were detected on two-dimensional electrophoresis gels, and 6 differentially expressed protein spots between 10 and 170 kDa were identified. Pathogenicity factors including S-adenosylmethionine synthetase, pyruvate dehydrogenase, glutathione synthetase, UDP-galactose-4-epimerase, acetate kinase A and elongation factor tu (EF-Tu). In validation of the pathogenicity factor, we used western blotting to show that Klebsiella pneumonia had higher (EF-Tu) expression when they accompanied by leukopenia rather than leukocytosis. Thus, we report 6 pathogenicity factors of leukopenia caused by Klebsiella pneumonia bacteremia, including 5 housekeeping enzymes and EF-Tu. We suggest EF-Tu could be a potential pathogenicity factor for leukopenia caused by Klebsiella pneumonia.

  10. mcr-1 Colistin Resistance in ESBL-Producing Klebsiella pneumoniae, France

    PubMed Central

    Maillet, Mylène; Pavese, Patricia; Francony, Gilles; Brion, Jean-Paul; Mallaret, Marie-Reine; Bonnet, Richard; Robin, Frédéric; Beyrouthy, Racha; Maurin, Max

    2017-01-01

    We report intestinal carriage of an extended-spectrum β-lactamase−producing Klebsiella pneumoniae strain with high-level resistance to colistin (MIC 24 mg/L) in a patient in France who had been hospitalized for fungal meningitis. The strain had the mcr-1 plasmid gene and an inactivated mgrB gene, which are associated with colistin resistance. PMID:28418313

  11. Intestinal Decontamination of Multidrug-resistant Klebsiella pneumoniae After Recurrent Infections in an Immunocompromised Host

    PubMed Central

    Kronman, Matthew P.; Zerr, Danielle M.; Qin, Xuan; Englund, Janet; Cornell, Cathy; Sanders, Jean E.; Myers, Jeffrey; Rayar, Jaipreet; Berry, Jessica E.; Adler, Amanda L.; Weissman, Scott J.

    2014-01-01

    Multidrug-resistant (MDR) Enterobacteriaceae infections are associated with increased morbidity. We describe a 20-year-old hematopoietic cell transplantation recipient with recurrent MDR Klebsiella pneumoniae infection, prolonged intestinal colonization, and subsequent intestinal decontamination. Further study should evaluate stool surveillance, molecular typing, and fecal microbiota transplantation for patients with intestinal MDR Enterobacteriaceae carriage. PMID:25041704

  12. Comparison of in vitro activities of meropenem productions on Klebsiella pneumoniae isolated from hospitalized patients.

    PubMed

    Akhi, Mohammad Taghi; Asghari, Babak; Nahaei, Mohammed Reza; Memar, Mohammad Yousef; Lari, Abdolaziz Rastegar; Naghili, Behrooz; Pirzadeh, Tahereh

    2014-01-01

    Antimicrobial activities of meropenem products on Klebsiella pneumoniae isolates were determined. 212 non-duplicated Klebsiella pneumoniae isolates were examined for in vitro meropenem susceptibility test by using the following disks, which were made from Meronem (AstraZeneca, UK), Exipenem (Exir, Iran) and Meroxan (DAANA, Iran) powders. MIC50 and MIC90 for meropenem antibiotics were determined. Meronem had good activities against most isolates of Klebsiella pneumoniae, and only a few strains had a rather high MIC. Exipenem and Meroxan showed a similar activity with Meronem. Regarding the comparison of two internal generic meropenem products with the external Meronem product have shown that they are equivalents in terms of microbiological activity, as measured using the disk diffusion and MIC. In developing countries, we suggested preparing disks with antibiotic powders that can be an equivalent function in microbiological activity with standard disks. In addition, since it demonstrated significant antimicrobial activity against the Klebsiella pneumoniae. For use of Exipenem and Meroxan in vivo, it would be better to perform additional testing (activity against different species, stability etc.).

  13. Non-clostridial gas gangrene caused by Klebsiella pneumoniae: a case report.

    PubMed

    Li, C M; Chen, P L; Ho, Y R

    2001-01-01

    A 45-y-old man was hospitalized due to pain and swelling of the right leg for 3 d. Bullae developed with gas formation involving multiple compartments of the entire limb 46 h later. Klebsiella pneumoniae was recovered from blood and surgical specimens. The patient died on Day 8 despite amputation and antibiotic therapy.

  14. Cerebellar abscess and meningitis, caused by Shewanella putrefaciens and Klebsiella pneumoniae, associated with chronic otitis media.

    PubMed

    Yilmaz, Gurdal; Aydin, Kemalettin; Bektas, Devrim; Caylan, Rahmet; Caylan, Refik; Koksal, Iftihar

    2007-11-01

    Shewanella putrefaciens is a facultatively anaerobic, non-motile, Gram-negative, non-fermentative bacterium. It is found in various environments and has been isolated worldwide. S. putrefaciens is a rare cause of brain abscesses and meningitis. This is a case report of a cerebellar abscess and meningitis caused by Shewanella putrefaciens and Klebsiella pneumoniae in a river trap fisherman.

  15. [Predominance and evolution of BLA(SHV) genes in Tunisian hospital isolates of Klebsiella pneumoniae].

    PubMed

    Ben-Hamouda, T; Ben-Mahrez, K

    2005-01-01

    Extended-spectrum beta-lactamases (ESBLs)produced by clinical strains of Klebsiella pneumoniae were investigated, using isoelectric-focusing and DNA amplification followed by sequencing. A predominance of SHV variants was found. Sequencing identified the genes for the SHV-2a and -12 enzymes, suggesting direct evolution of SHV-12 from SHV-2a.

  16. Genome Sequence of OXA-48 Carbapenemase-Producing Klebsiella pneumoniae KpO3210

    PubMed Central

    Wesselink, Jan-Jaap; López-Camacho, Elena; de la Peña, Santiago; Ramos-Ruiz, Ricardo; Ruiz-Carrascoso, Guillermo; Lusa-Bernal, Silvia; Fernández-Soria, Victor M.; Gómez-Gil, Rosa

    2012-01-01

    Klebsiella pneumoniae KpO3210 is a OXA-48 carbapenemase-producing isolate obtained from a blood culture in the context of an outbreak in Hospital Universitario La Paz (Madrid, Spain) in 2010. It belongs to the major clone detected during the outbreak and is resistant to all beta-lactams and to several other antibiotics. PMID:23209233

  17. Epidemiological typing of multidrug-resistant Klebsiella pneumoniae, which causes paediatric ventilator-associated pneumonia in Egypt.

    PubMed

    Mohamed, Eman R; Aly, Sherine A; Halby, Hamada M; Ahmed, Shabaan H; Zakaria, Amira M; El-Asheer, Osama M

    2017-05-01

    Multidrug-resistant Klebsiella pneumoniae is a common nosocomial pathogen that plays an important role in ventilator-associated pneumonia (VAP). This study aimed to define the clonal relatedness of K. pneumoniae strains isolated from paediatric VAP in addition to those isolated from environmental samples. This study included 19 clinical and 4 environmental K. pneumoniae isolates recovered from the paediatric intensive care unit (PICU) in Assiut University Children's Hospital. The K. pneumoniae isolates were confirmed by biotyping using API strips and subjected to antimicrobial susceptibility testing. The genes coding K1 and K2 capsular types were detected by PCR. The clonal relationships between the K. pneumoniae isolates were determined by pulsed-field gel electrophoresis (PFGE). Ten resistotypes were detected among all the K. pneumoniae isolates, while PFGE identified seventeen K. pneumoniae pulsotypes. Similar PFGE patterns were found between environmental and clinical isolates and between isolates recovered from different patients, suggesting the circulation of K. pneumoniae pathogens in the PICU and the role of the environment in the spread of infection. No correlation was found between the resistotypes and pulsotypes of the K. pneumoniae isolates. PFGE showed higher discriminatory power for the typing of nosocomial K. pneumoniae [Simpson's diversity index (DI)=0.96] than resistotyping (DI=0.72). As far as we know, this is the first report of the isolation of the same multidrug-resistant (MDR) K. pneumoniae pulsotype from patients and environmental samples in the same hospital ward in Egypt. This study provides a step on the way to understanding the genotyping and epidemiology of MDR K. pneumoniae for enhanced prevention of bacterial transmission.

  18. Use of bacteriophage Mu to isolate deletions in the his-nif region of Klebsiella pneumoniae.

    PubMed Central

    Bachhuber, M; Brill, W J; Howe, M M

    1976-01-01

    Klebsiella pneumoniae M5a1 is naturally resistant to infection by bacteriophage Mu. Mutants of K. pneumoniae sensitive to Mu infection were isolated and found to support both lytic and lysogenic development of Mu. K. pneumoniae lysogens containing a heat-inducible Mu prophage integrated in his were isolated. Strains carrying deletions extending from his into nif were obtained after heat treatment of these lysogens. Such deletions should be useful for determining the map order and cistronic organization of the nif genes. PMID:791929

  19. Pneumocephalus as a complication of multidrug-resistant Klebsiella pneumoniae meningitis.

    PubMed

    Sreejith, P; Vishad, V; Pappachan, Joseph M; Laly, D C; Jayaprakash, R; Ranjith, V T

    2008-03-01

    Pneumocephalus implies air inside the cranial vault, which usually results from cranio-facial trauma. Occasionally, meningitis caused by gas-forming organisms can result in pneumocephalus. Klebsiella pneumoniae meningitis can, on rare occasions, cause pneumocephalus as a complication. The drug of choice for K. pneumoniae meningitis is a third-generation cephalosporin, and resistance to these drugs is unusual. We report a case of multidrug-resistant K. pneumoniae meningitis resulting from chronic suppurative otitis media, which was later complicated by pneumocephalus. The patient was successfully managed with meropenam and amikacin, the only antibiotics to which these bacilli showed no resistance.

  20. Identification of putative plant pathogenic determinants from a draft genome sequence of an opportunistic klebsiella pneumoniae strain

    USDA-ARS?s Scientific Manuscript database

    Klebsiella pneumoniae has been known historically as a causal agent of bacterial pneumonia. More recently, K. pneumoniaerepresentatives have been shown to have a broad ecological distribution and are recognized nitrogen-fixers. Previously, we demonstrated the capacity of K. pneumoniae strain Kp 5-1R...

  1. Carbapenemase-Producing Klebsiella pneumoniae in Romania: A Six-Month Survey.

    PubMed

    Lixandru, Brandusa Elena; Cotar, Ani Ioana; Straut, Monica; Usein, Codruta Romanita; Cristea, Dana; Ciontea, Simona; Tatu-Chitoiu, Dorina; Codita, Irina; Rafila, Alexandru; Nica, Maria; Buzea, Mariana; Baicus, Anda; Ghita, Mihaela Camelia; Nistor, Irina; Tuchiluş, Cristina; Indreas, Marina; Antohe, Felicia; Glasner, Corinna; Grundmann, Hajo; Jasir, Aftab; Damian, Maria

    2015-01-01

    This study presents the first characterization of carbapenem-non-susceptible Klebsiella pneumoniae isolates by means of a structured six-month survey performed in Romania as part of an Europe-wide investigation. Klebsiella pneumoniae clinical isolates from different anatomical sites were tested for antibiotic susceptibility by phenotypic methods and confirmed by PCR for the presence of four carbapenemase genes. Genome macrorestriction fingerprinting with XbaI was used to analyze the relatedness of carbapenemase-producing Klebsiella pneumoniae isolates collected from eight hospitals. Among 75 non-susceptible isolates, 65 were carbapenemase producers. The most frequently identified genotype was OXA-48 (n = 51 isolates), eight isolates were positive for blaNDM-1 gene, four had the blaKPC-2 gene, whereas two were positive for blaVIM-1. The analysis of PFGE profiles of OXA-48 and NDM-1 producing K. pneumoniae suggests inter-hospitals and regional transmission of epidemic clones. This study presents the first description of K. pneumoniae strains harbouring blaKPC-2 and blaVIM-1 genes in Romania. The results of this study highlight the urgent need for the strengthening of hospital infection control measures in Romania in order to curb the further spread of the antibiotic resistance.

  2. Fatal Klebsiella pneumoniae meningitis in a patient with diabetes mellitus and Hansen's disease.

    PubMed

    Gopal, Vani; Mangaiyarkarasi, T; Gopal, R

    2014-01-01

    Klebsiella is a Gram-negative bacterium that causes different types of health care-associated infections including pneumonia, bloodstream infections, surgical site infections and meningitis. We report here a case of Klebsiella pneumoniae meningitis in a patient with diabetes mellitus and Hansen's disease. A middle-aged man with a known case of diabetes mellitus and Hansen's disease presented with the complaints of blurred vision in the left eye and the patient was found to have cataract. Patient was operated for cataract and Intraocular lens implanted. Patient developed headache and vomiting on the 4th post-operative day. Lumbar puncture was carried out and gram stain of cerebrospinal fluid showed Gram-negative bacilli in the direct smear and culture yielded a heavy growth of K. pneumoniae. The patient was treated with antimicrobials according to the susceptibility pattern. He initially showed improvement but later on developed altered sensorium and hypotension. Patient succumbed to infection in spite of all medical attention.

  3. Pathogenicity of Aeromonas hydrophila, Klebsiella pneumoniae, and Proteus mirabilis to brown tree frogs (Litoria ewingii).

    PubMed

    Schadich, Ermin; Cole, Anthony L J

    2010-04-01

    Bacterial dermatosepticemia, a systemic infectious bacterial disease of frogs, can be caused by several opportunistic gram-negative bacterial species including Aeromonas hydrophila, Chryseobacterium indologenes, Chryseobacterium meningosepticum, Citrobacter freundii, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, and Serratia liquifaciens. Here we determined the pathogenicity of 3 bacterial species (Aeromonas hydrophila, Klebsiella pneumoniae, and Proteus mirabilis) associated with an outbreak of fatal dermatosepticemia in New Zealand Litoria ewingii frogs. A bath challenge method was used to expose test frogs to individual bacterial species (2 x 10(7) cfu/mL in pond water); control frogs were exposed to uninfected pond water. None of the control frogs or those exposed to A. hydrophila or P. mirabilis showed any morbidity or mortality. Morbidity and mortality was 40% among frogs exposed to K. pneumonia, and the organism was reisolated from the hearts, spleens, and livers of affected animals.

  4. Pathogenicity of Aeromonas hydrophila, Klebsiella pneumoniae, and Proteus mirabilis to Brown Tree Frogs (Litoria ewingii)

    PubMed Central

    Schadich, Ermin; Cole, Anthony LJ

    2010-01-01

    Bacterial dermatosepticemia, a systemic infectious bacterial disease of frogs, can be caused by several opportunistic gram-negative bacterial species including Aeromonas hydrophila, Chryseobacterium indologenes, Chryseobacterium meningosepticum, Citrobacter freundii, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, and Serratia liquifaciens. Here we determined the pathogenicity of 3 bacterial species (Aeromonas hydrophila, Klebsiella pneumoniae, and Proteus mirabilis) associated with an outbreak of fatal dermatosepticemia in New Zealand Litoria ewingii frogs. A bath challenge method was used to expose test frogs to individual bacterial species (2 × 107 cfu/mL in pond water); control frogs were exposed to uninfected pond water. None of the control frogs or those exposed to A. hydrophila or P. mirabilis showed any morbidity or mortality. Morbidity and mortality was 40% among frogs exposed to K. pneumonia, and the organism was reisolated from the hearts, spleens, and livers of affected animals. PMID:20412685

  5. Ventilator-associated pneumonia caused by colistin-resistant KPC-producing Klebsiella pneumoniae: a case report and literature review.

    PubMed

    Viaggi, Bruno; Sbrana, Francesco; Malacarne, Paolo; Tascini, Carlo

    2015-05-01

    Klebsiella pneumoniae producing KPC-type carbapenemase causes severe nosocomial infection at a high mortality rate. Nosocomial pneumonia in particular is associated with high mortality, likely due to the unfavorable pulmonary pharmacokinetics of the antibiotics used against this agent. Therefore, early and accurate microbiological identification and susceptibility evaluation are crucial in order to optimize antibiotic therapy. We report a case of ventilator-associated pneumonia caused by colistin-resistant K. pneumoniae producing KPC-type carbapenemase treated using a carbapenem-sparing therapy and tailored according to the serum procalcitonin concentration in order to limit the duration of antibiotic therapy. Copyright © 2015 The Japanese Respiratory Society. Published by Elsevier B.V. All rights reserved.

  6. Acute lung inflammation in Klebsiella pneumoniae B5055-induced pneumonia and sepsis in BALB/c mice: a comparative study.

    PubMed

    Kumar, Vijay; Chhibber, Sanjay

    2011-10-01

    Lungs play an important role in the body's defense against a variety of pathogens, but this network of immune system-mediated defense can be deregulated during acute pulmonary infections. The present study compares acute lung inflammation occurring during Klebsiella pneumoniae B5055-induced pneumonia and sepsis in BALB/c mice. Pneumonia was induced by intranasal instillation of bacteria (10(4) cfu), while sepsis was developed by placing the fibrin-thrombin clot containing known amount of bacteria (10(2) cfu) into the peritoneal cavity of animals. Mice with sepsis showed 100% mortality within five post-infection days, whereas all the animals with pneumonia survived. In animals suffering from K. pneumoniae B5055-induced pneumonia, all the inflammatory parameters (TNF-α, IL-1α, MPO, MDA, and NO) were found to be maximum till third post-infection day, after that, a decline was observed, whereas in septic animals, all the above-mentioned markers of inflammation kept on increasing. Histopathological study showed presence of alternatively activated alveolar macrophages (or foam cells) in lungs of mice with pneumonia after third post-infection day, which might have contributed to the induction of resolution of inflammation, but no such observation was made in lungs of septic mice. Hence, during pneumonia, controlled activation of macrophages may lead to resolution of inflammation.

  7. A Serendipitous Mutation Reveals the Severe Virulence Defect of a Klebsiella pneumoniae fepB Mutant

    PubMed Central

    Palacios, Michelle; Broberg, Christopher A.; Walker, Kimberly A.

    2017-01-01

    ABSTRACT Klebsiella pneumoniae is considered a significant public health threat because of the emergence of multidrug-resistant strains and the challenge associated with treating life-threatening infections. Capsule, siderophores, and adhesins have been implicated as virulence determinants of K. pneumoniae, yet we lack a clear understanding of how this pathogen causes disease. In a previous screen for virulence genes, we identified a potential new virulence locus and constructed a mutant (smr) with this locus deleted. In this study, we characterize the smr mutant and show that this mutation renders K. pneumoniae avirulent in a pneumonia model of infection. The smr mutant was expected to have a deletion of three genes, but subsequent genome sequencing indicated that a much larger deletion had occurred. Further analysis of the deleted region indicated that the virulence defect of the smr mutant could be attributed to the loss of FepB, a periplasmic protein required for import of the siderophore enterobactin. Interestingly, a ΔfepB mutant was more attenuated than a mutant unable to synthesize enterobactin, suggesting that additional processes are affected. As FepB is highly conserved among the members of the family Enterobacteriaceae, therapeutic targeting of FepB may be useful for the treatment of Klebsiella and other bacterial infections. IMPORTANCE In addition to having a reputation as the causative agent of several types of hospital-acquired infections, Klebsiella pneumoniae has gained widespread attention as a pathogen with a propensity for acquiring antibiotic resistance. It is capable of causing a range of infections, including urinary tract infections, pneumonia, and sepsis. Because of the rapid emergence of carbapenem resistance among Klebsiella strains, there is a dire need for a better understanding of virulence mechanisms and identification of new drug targets. Here, we identify the periplasmic transporter FepB as one such potential target. PMID

  8. Susceptibility of Klebsiella pneumoniae on coriander leaves to liquid- and vapor-phase ethanol.

    PubMed

    Krusong, Warawut; Pornpukdeewatana, Soisuda; Teerarak, Montinee

    2016-05-01

    The bio-control of ethanol on Klebsiella pneumoniae on fresh coriander leaves for significantly reducing consumer health risk was investigated. Washed and sterilized leaves of coriander were inoculated with K. pneumoniae cultured in Trypticase Soy broth. Susceptibility of the K. pneumoniae to liquid- and evaporated vapor-phase ethanol (EVE) was then examined in vitro Complete inhibition of K. pneumoniae was found with 18% (v/v) liquid ethanol. Exposure for 15 min to EVE (9.00 ± 0.8 mmol L(-1)) completely destroyed K. pneumoniae (4.04 ± 0.02 log CFU/ml) spread on Mueller Hilton agar at 30 ± 2°C. The effect of EVE with and without evaporated water vapor (EWV) on the susceptibility of K. pneumoniae on fresh coriander leaves was examined. While exposure to EVE affected the survival of K. pneumoniae, the degree of reduction depended on both the inoculation level and the EWV. Complete reduction of K. pneumoniae was achieved for the low inoculation level by EVE alone (37 ± 2% relative humidity; RH) but susceptibility was reduced with EWV (high RH; 80 ± 2%). Scanning electron microscope (SEM) images of inoculated coriander leaves confirm the effects of EVE in reducing levels of K. pneumoniae Exposure to EVE alone proved an effective bio-control for K. pneumoniae on fresh coriander leaves.

  9. Anti-inflammatory effects of inhaled nitric oxide are optimized at lower oxygen concentration in experimental Klebsiella pneumoniae pneumonia.

    PubMed

    Sun, Z; Sun, B; Wang, X; Wang, W; Zhu, L

    2006-10-01

    To evaluate whether antiinflammatory effects of inhaled nitric oxide (NO) in experimental Klebsiella pneumoniae pneumonia may be affected by oxygen levels in association with cytokine response and NO synthase (NOS) activity. Healthy adult rats were intratracheally instilled with live Klebsiella pneumoniae to induce pneumonia (P) or with sterile saline as a control (C) group. Group C was allocated to room air (CA) and inhaled NO (CNO, NO = 20 parts per million) groups. Group P was divided into 6 groups (n = 8 - 10) exposed to room air (PA), inhaled NO (PNO), low oxygen (40%, PLO), inhaled NO and low oxygen (PLONO), high oxygen (100%, PHO), and inhaled NO and high oxygen (98%, PHONO). After 24 h exposure, intraalveolar cells, expression of proinflammatory cytokines, nuclear transcription factor (NF-kappaB), myeloperoxidase (MPO) and NOS activities, characteristic of lung inflammatory mechanisms, were measured. All the pneumonia groups had pneumonia whereas CA and CNO had no or mild lung inflammation. Compared to the PA, significantly decreased recruitment of alveolar neutrophils was found in the PNO, along with lower NF-kappaB and MPO activity, tumor necrosis factor-alpha (TNF-alpha) and intercellular adhesion molecule-1 (ICAM-1). Similar trends were found between the PLONO and PLO in alleviation of lung inflammation. However, PHONO significantly enhanced recruitment of alveolar neutrophils and pulmonary MPO activity associated with increased TNF-alpha. Inhaled NO improved bacterial clearance, reduced total proteins in alveoli, and ICAM-1 expression irrespective of oxygen levels. Inhaled NO and/or low and high oxygen partially restored constitutive NOS activity whereas high oxygen or together with inhaled NO depressed inducible NOS activity. The overall beneficial effects of inhaled NO in anti-inflammation in the mature lungs with K. pneumoniae pneumonia are in favor of a combination with lower level of oxygen, which warrants clinical verification in the treatment of

  10. X-linked agammaglobulinemia combined with juvenile idiopathic arthritis and invasive Klebsiella pneumoniae polyarticular septic arthritis.

    PubMed

    Zhu, Zaihua; Kang, Yuli; Lin, Zhenlang; Huang, Yanjing; Lv, Huoyang; Li, Yasong

    2015-02-01

    X-linked agammaglobulinemia (XLA) is a primary immunodeficiency disease caused by mutations in the Bruton's tyrosine kinase (BTK) gene. XLA can also present in combination with juvenile idiopathic arthritis (JIA), the major chronic rheumatologic disease in children. We report herein the first known case of a juvenile patient diagnosed with XLA combined with JIA that later developed into invasive Klebsiella pneumoniae polyarticular septic polyarthritis. An additional comprehensive review of XLA combined with JIA and invasive K. pneumoniae septic arthritis is also presented. XLA was identified by the detection of BTK mutations while the diagnosis of JIA was established by clinical and laboratory assessments. Septic arthritis caused by invasive K. pneumoniae was confirmed by culturing of the synovia and gene detection of the isolates. Invasive K. pneumoniae infections can not only result in liver abscesses but also septic arthritis, although this is rare. XLA combined with JIA may contribute to invasive K. pneumoniae infection.

  11. Potential pathogens in the environment: Klebsiella pneumoniae, a taxonomic and ecological enigma.

    PubMed

    Brown, C; Seidler, R J

    1973-06-01

    A nitrogen-deficient medium and m-Endo agar were employed in the isolation of members of the tribe Klebsielleae from surfaces of vegetables and seeds. With m-Endo agar at an incubation temperature of 37 C, nearly 50% of the vegetables and seven out of seven seed samples yielded organisms which biochemically and serologically were identified as Klebsiella pneumoniae, Viable counts were generally in the range of 10(3) cells per g of vegetable peel or seed. Organisms classified as K. pneumoniae exhibited seven different IMViC patterns, with the --++, ++++, and -+++ patterns most common. Seven of the eleven K. pneumoniae serotypes encountered have previously been isolated from human urinary tract and other infections. Fifty percent of the 40 K. pneumoniae examined exhibited positive acetylene-reducing activity, i.e., they possessed the capability for fixing N(2). Vegetables containing K. pneumoniae may constitute a potential reservoir for human nosocomial genitourinary or other infections.

  12. Alcohol-associated intestinal dysbiosis impairs pulmonary host defense against Klebsiella pneumoniae.

    PubMed

    Samuelson, Derrick R; Shellito, Judd E; Maffei, Vincent J; Tague, Eric D; Campagna, Shawn R; Blanchard, Eugene E; Luo, Meng; Taylor, Christopher M; Ronis, Martin J J; Molina, Patricia E; Welsh, David A

    2017-06-01

    Chronic alcohol consumption perturbs the normal intestinal microbial communities (dysbiosis). To investigate the relationship between alcohol-mediated dysbiosis and pulmonary host defense we developed a fecal adoptive transfer model, which allows us to investigate the impact of alcohol-induced gut dysbiosis on host immune response to an infectious challenge at a distal organ, independent of prevailing alcohol use. Male C57BL/6 mice were treated with a cocktail of antibiotics (ampicillin, gentamicin, neomycin, vancomycin, and metronidazole) via daily gavage for two weeks. A separate group of animals was fed a chronic alcohol (or isocaloric dextrose pair-fed controls) liquid diet for 10 days. Microbiota-depleted mice were recolonized with intestinal microbiota from alcohol-fed or pair-fed (control) animals. Following recolonization groups of mice were sacrificed prior to and 48 hrs. post respiratory infection with Klebsiella pneumoniae. Klebsiella lung burden, lung immunology and inflammation, as well as intestinal immunology, inflammation, and barrier damage were examined. Results showed that alcohol-associated susceptibility to K. pneumoniae is, in part, mediated by gut dysbiosis, as alcohol-naïve animals recolonized with a microbiota isolated from alcohol-fed mice had an increased respiratory burden of K. pneumoniae compared to mice recolonized with a control microbiota. The increased susceptibility in alcohol-dysbiosis recolonized animals was associated with an increase in pulmonary inflammatory cytokines, and a decrease in the number of CD4+ and CD8+ T-cells in the lung following Klebsiella infection but an increase in T-cell counts in the intestinal tract following Klebsiella infection, suggesting intestinal T-cell sequestration as a factor in impaired lung host defense. Mice recolonized with an alcohol-dysbiotic microbiota also had increased intestinal damage as measured by increased levels of serum intestinal fatty acid binding protein. Collectively, these

  13. Alcohol-associated intestinal dysbiosis impairs pulmonary host defense against Klebsiella pneumoniae

    PubMed Central

    Campagna, Shawn R.; Blanchard, Eugene E.; Ronis, Martin J. J.

    2017-01-01

    Chronic alcohol consumption perturbs the normal intestinal microbial communities (dysbiosis). To investigate the relationship between alcohol-mediated dysbiosis and pulmonary host defense we developed a fecal adoptive transfer model, which allows us to investigate the impact of alcohol-induced gut dysbiosis on host immune response to an infectious challenge at a distal organ, independent of prevailing alcohol use. Male C57BL/6 mice were treated with a cocktail of antibiotics (ampicillin, gentamicin, neomycin, vancomycin, and metronidazole) via daily gavage for two weeks. A separate group of animals was fed a chronic alcohol (or isocaloric dextrose pair-fed controls) liquid diet for 10 days. Microbiota-depleted mice were recolonized with intestinal microbiota from alcohol-fed or pair-fed (control) animals. Following recolonization groups of mice were sacrificed prior to and 48 hrs. post respiratory infection with Klebsiella pneumoniae. Klebsiella lung burden, lung immunology and inflammation, as well as intestinal immunology, inflammation, and barrier damage were examined. Results showed that alcohol-associated susceptibility to K. pneumoniae is, in part, mediated by gut dysbiosis, as alcohol-naïve animals recolonized with a microbiota isolated from alcohol-fed mice had an increased respiratory burden of K. pneumoniae compared to mice recolonized with a control microbiota. The increased susceptibility in alcohol-dysbiosis recolonized animals was associated with an increase in pulmonary inflammatory cytokines, and a decrease in the number of CD4+ and CD8+ T-cells in the lung following Klebsiella infection but an increase in T-cell counts in the intestinal tract following Klebsiella infection, suggesting intestinal T-cell sequestration as a factor in impaired lung host defense. Mice recolonized with an alcohol-dysbiotic microbiota also had increased intestinal damage as measured by increased levels of serum intestinal fatty acid binding protein. Collectively, these

  14. Quantifying the clinical virulence of Klebsiella pneumoniae producing carbapenemase Klebsiella pneumoniae with a Galleria mellonella model and a pilot study to translate to patient outcomes.

    PubMed

    McLaughlin, Milena M; Advincula, M Renee; Malczynski, Michael; Barajas, Grace; Qi, Chao; Scheetz, Marc H

    2014-01-15

    Previous studies may have overestimated morbidity and mortality due to Klebsiella pneumoniae producing carbapenemase (KPC) Klebsiella pneumoniae infections because of difficulties in modeling patient comorbidities. This pilot study sought to evaluate KPC virulence by combining clinical and Galleria mellonella models in patients with K. pneumoniae blood stream infections (BSIs). G. mellonella were inoculated using KPC(+) and KPC(-) isolates from these patients. Extent and rapidity of insect mortality was analyzed. Patients were stratified by KPC BSI status. Clinical outcomes of mortality and length of stay post-infection for survivors (LOS) were analyzed. Median virulence scores calculated from the insect studies were imputed in the clinical model. The in-vivo model revealed greater mortality in KPC(-) isolates (p < 0.001). Fifteen patients with KPC(+) BSI were matched with 60 patients with KPC(-) BSI. Hospital mortality was greater in the KPC(+) group versus the KPC(-) group (OR 3.79, 95% CI 1.00 - 14.34). LOS was longer in the KPC(+) group (p < 0.01). Conversely the virulence score attenuated the association between KPC(+) status and mortality and LOS in the final translational models. KPC(+) status was associated with decreased virulence in GM. Opposite findings were observed in patients. This pilot study demonstrates that measured virulence from GM may differ from human estimates of virulence.

  15. An Outbreak of Infections Caused by a Klebsiella pneumoniae ST11 Clone Coproducing Klebsiella pneumoniae Carbapenemase-2 and RmtB in a Chinese Teaching Hospital

    PubMed Central

    Li, Jun; Zou, Ming-Xiang; Wang, Hai-Chen; Dou, Qing-Ya; Hu, Yong-Mei; Yan, Qun; Liu, Wen-En

    2016-01-01

    Background: Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae bacteria, which cause serious disease outbreaks worldwide, was rarely detected in Xiangya Hospital, prior to an outbreak that occurred from August 4, 2014, to March 17, 2015. The aim of this study was to analyze the epidemiology and molecular characteristics of the K. pneumoniae strains isolated during the outbreak. Methods: Nonduplicate carbapenem-resistant K. pneumoniae isolates were screened for blaKPC-2 and multiple other resistance determinants using polymerase chain reaction. Subsequent studies included pulsed-field gel electrophoresis (PFGE), multilocus sequence typing, analysis of plasmids, and genetic organization of blaKPC-2 locus. Results: Seventeen blaKPC-2-positive K. pneumoniae were identified. A wide range of resistant determinants was detected. Most isolates (88.2%) coharbored blaKPC-2 and rmtB in addition to other resistance genes, including blaSHV-1, blaTEM-1, and aac(3)-IIa. The blaKPC-2 and rmtB genes were located on the conjugative IncFIB-type plasmid. Genetic organization of blaKPC-2 locusin most strains was consistent with that of the plasmid pKP048. Four types (A1, A2, A3, and B) were detected by PFGE, and Type A1, an ST11, was the predominant PFGE type. A novel K. pneumoniae sequence type (ST1883) related to ST11 was discovered. Conclusions: These isolates in our study appeared to be clonal and ST11 K. pneumoniae was the predominant clone attributed to the outbreak. Coharbing of blaKPC-2 and rmtB, which were located on a transferable plasmid, in clinical K. pneumoniae isolates may lead to the emergence of a new pattern of drug resistance. PMID:27569227

  16. Pneumonia due to pandemic (H1N1) 2009 influenza virus and Klebsiella pneumoniae capsular serotype K16 in a patient with nasopharyngeal cancer.

    PubMed

    Lai, Chih-Cheng; Lee, Pei-Lin; Tan, Che-Kim; Huang, Yu-Tsung; Kao, Chiang-Lian; Wang, Jin-Town; Hsueh, Po-Ren

    2012-10-01

    Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus and group A Streptoccocus, but no Klebsiella pneumoniae were responsible for bacterial coinfections during the 2009 and previous influenza pandemics. We hereby report a case with concurrent bacteremic pneumonia due to an unusual capsular serotype K16 K. pneumoniae and pandemic (H1N1) 2009 influenza in a patient with nasopharyngeal cancer. Such a coinfection has not previously been described. Copyright © 2012. Published by Elsevier B.V.

  17. Dissemination of extensively drug-resistant and KPC-2 producing Klebsiella pneumoniae isolated from bloodstream infections.

    PubMed

    Zhao, Feng; Zhang, Jun; Fu, Ying; Ruan, Zhi; Xie, Xinyou

    2015-09-27

    Bloodstream infections (BSIs) are serious diseases associated with high mortality, especially when caused by extensively drug-resistant (XDR) Klebsiella pneumoniae. The prevalence and pandemic strains of extended-spectrum beta-lactamase (ESBL)-producing K. pneumoniae isolated from blood cultures of patients with BSIs were determined at Sir Run Run Shaw Hospital, China. A total of 24 XDR K. pneumoniae were isolated from blood cultures, and the clinical data of the patients were analyzed retrospectively. Bacterial species identification and antimicrobial susceptibility testing were performed using VITEK2 and E-test methods, respectively. Common ESBL-resistant genes were amplified and sequenced after the validation of a modified Hodge test. Strain homology was also analyzed by pulsed-field gel electrophoresis (PFGE). All of the isolates were resistant to 10 antimicrobial agents. Several strains showed partial sensitivity to aminoglycosides, but all showed sensitivity to polymyxin and tigecycline. All strains were Klebsiella pneumoniae carbapenemase (KPC-2) producing, and carried two or three ESBL-resistant genes, which belonged to 13 PFGE clones (A-M). The overall mortality rate in patients was as high as 29.2%. KPC-2-producing K. pneumoniae BSIs are associated with high mortality rates. Our observations suggest that KPC-2 and ESBL-producing K. pneumoniae might be responsible for the clonal dissemination of extensively drug-resistant isolates in our hospital.

  18. Novel screening assay for in vivo selection of Klebsiella pneumoniae genes promoting gastrointestinal colonisation

    PubMed Central

    2012-01-01

    Background Klebsiella pneumoniae is an important opportunistic pathogen causing pneumonia, sepsis and urinary tract infections. Colonisation of the gastrointestinal (GI) tract is a key step in the development of infections; yet the specific factors important for K. pneumoniae to colonize and reside in the GI tract of the host are largely unknown. To identify K. pneumoniae genes promoting GI colonisation, a novel genomic-library-based approach was employed. Results Screening of a K. pneumoniae C3091 genomic library, expressed in E. coli strain EPI100, in a mouse model of GI colonisation led to the positive selection of five clones containing genes promoting persistent colonisation of the mouse GI tract. These included genes encoding the global response regulator ArcA; GalET of the galactose operon; and a cluster of two putative membrane-associated proteins of unknown function. Both ArcA and GalET are known to be involved in metabolic pathways in Klebsiella but may have additional biological actions beneficial to the pathogen. In support of this, GalET was found to confer decreased bile salt sensitivity to EPI100. Conclusions The present work establishes the use of genomic-library-based in vivo screening assays as a valuable tool for identification and characterization of virulence factors in K. pneumoniae and other bacterial pathogens. PMID:22967317

  19. KlebSeq, a Diagnostic Tool for Surveillance, Detection, and Monitoring of Klebsiella pneumoniae

    PubMed Central

    Lemmer, Darrin; Sahl, Jason W.; Pearson, Talima; Driebe, Elizabeth M.; Wojack, Bette; Saubolle, Michael A.; Engelthaler, David M.; Keim, Paul

    2016-01-01

    Health care-acquired infections (HAIs) kill tens of thousands of people each year and add significantly to health care costs. Multidrug-resistant and epidemic strains are a large proportion of HAI agents, and multidrug-resistant strains of Klebsiella pneumoniae, a leading HAI agent, have caused an urgent public health crisis. In the health care environment, patient colonization by K. pneumoniae precedes infection, and transmission via colonization leads to outbreaks. Periodic patient screening for K. pneumoniae colonization has the potential to curb the number of HAIs. In this report, we describe the design and validation of KlebSeq, a highly informative screening tool that detects Klebsiella species and identifies clinically important strains and characteristics by using highly multiplexed amplicon sequencing without a live-culturing step. We demonstrate the utility of this tool on several complex specimen types, including urine, wound swabs and tissue, and several types of respiratory and fecal specimens, showing K. pneumoniae species and clonal group identification and antimicrobial resistance and virulence profiling, including capsule typing. Use of this amplicon sequencing tool to screen patients for Klebsiella carriage could inform health care staff of the risk of infection and outbreak potential. KlebSeq also serves as a model for next-generation molecular tools for public health and health care, as expansion of this tool can be used for several other HAI agents or applications. PMID:27510832

  20. Effect of radiation processing in elimination of Klebsiella pneumoniae from food

    NASA Astrophysics Data System (ADS)

    Gautam, Raj Kamal; Nagar, Vandan; Shashidhar, Ravindranath

    2015-10-01

    Klebsiella pneumoniae has been considered as an important foodborne pathogen which causes severe infections that include meningitis, bronchitis, bacteremia, pneumonia, and urinary tract infections in humans and animals. It is well known to most clinicians as a cause of community-acquired bacterial pneumonia. Klebsiella is an opportunistic pathogen, that primarily attacks neonates, infants, elderly and immuno-compromised patients and therefore impose a serious, emerging public health hazard globally. Contaminated sprouts, vegetables, seafood and other animal meat products are considered as main sources of Klebsiella infection. In the current study, radiation sensitivity of K. pneumoniae MTCC 109 was determined in different food samples. The decimal reduction dose (D10) values of K. pneumoniae MTCC 109 in saline and nutrient broth at 0-4 °C were 0.116±0.009, 0.136±0.005 kGy, respectively. The mixed sprouts, fish and poultry samples were inoculated with K. pneumoniae MTCC 109 and exposed to gamma radiation to evaluate the effectiveness of radiation treatment in the elimination of K. pneumoniae. D10 values of K. pneumoniae in mixed sprouts, poultry and fish samples were found to be 0.142±0.009, 0.125±0.0004 and 0.277±0.012 kGy, respectively. Radiation treatment with a 1.5 kGy dose resulted in the complete elimination of 3.1±1.8×105 CFU/g of K. pneumoniae from these food samples. No recovery of K. pneumoniae was observed in the 1.5 kGy treated samples stored at 4 °C up to 12 days, even after enrichment and selective plating. This study shows that a 1.5 kGy dose of irradiation treatment could lead to the complete elimination of 3.1±1.8×105 CFU/g of K. pneumoniae from mixed sprouts, poultry and fish samples.

  1. Construction and expression of recombinant plasmids encoding type 1 fimbriae of a urinary Klebsiella pneumoniae isolate.

    PubMed Central

    Purcell, B K; Clegg, S

    1983-01-01

    The type 1 fimbriae of Klebsiella pneumoniae have been implicated as important virulence factors in mediating Klebsiella urinary infections. The chromosomally encoded fimbrial genes were cloned by a cosmid cloning technique. Further subcloning was performed with the cloning vehicles pBR322 and pACYC184, and a recombinant plasmid containing the fimbrial genes was constructed. After transformation by this plasmid, both Escherichia coli and Salmonella typhimurium were shown to express fimbriae which reacted with Klebsiella fimbrial antiserum. The approximate location of the relevant genes on the chimeric plasmid was determined by insertion of the transposable element Tn5. Hemagglutination-negative phenotypes were used to estimate the minimum size of the DNA fragment necessary to encode fimbrial biosynthesis and expression. The size of the coding region of this fragment was found to be 5.5 kilobase pairs. PMID:6132874

  2. Fulminating gas-forming psoas muscle abscess due to Klebsiella pneumoniae following a deep neck infection.

    PubMed

    Jang, T N; Juang, G D; Fung, C P

    1997-02-01

    Psoas muscle abscess due to Klebsiella pneumoniae infection is rare. We report a 55-year-old diabetic man who presented with progressive back pain of 1 month's duration. The patient had undergone surgical drainage for a deep neck infection with K. pneumoniae 43 days previously. On the present admission, physical examination revealed tenderness over the anterior upper aspect of both thighs, and computed tomography showed pneumoretroperitoneum dissecting the bilateral iliopsoas muscles. Parenteral administration of antibiotics was started immediately. Due to the patient's poor health status, we opted for repeated computed tomographic and sonographic-guided percutaneous drainage rather than surgical drainage. Blood and pus cultures revealed only K. pneumoniae. The patient recovered without significant sequelae. This report stresses the risk of metastatic infections caused by K. pneumoniae, especially in diabetic patients. Our experience suggests that repeated percutaneous drainage is feasible in cases of severe iliopsoas abscess, especially when risks associated with surgery are high.

  3. Septic arthritis subsequent to urosepsis caused by hypermucoviscous Klebsiella pneumoniae.

    PubMed

    Suzuki, Kei; Nakamura, Akiko; Enokiya, Tomoyuki; Iwashita, Yoshiaki; Tomatsu, Eri; Muraki, Yuichi; Kaneko, Toshihiro; Okuda, Masahiro; Katayama, Naoyuki; Imai, Hiroshi

    2013-01-01

    We herein report the first case of septic arthritis caused by rmpA-positive hypermucoviscous community-acquired K. pneumoniae that followed urosepsis in a 65-year-old Japanese woman. The patient responded well to drainage of the abscesses and treatment with cefazolin. Although this virulent phenotype of K. pneumoniae has been primarily reported in Hong Kong, we confirmed that 18/50 isolates obtained in our hospital over the past five years displayed the hypermucoviscous phenotype. Therefore, clinicians should consider the possibility of an increasing prevalence of rmpA-positive hypermucoviscous K. pneumoniae infection in Japan and be particularly vigilant for invasive clinical manifestations, even in patients with urinary tract infections.

  4. Multi-Drug-Resistant Klebsiella pneumoniae Pancreatitis: A New Challenge in a Serious Surgical Infection

    PubMed Central

    Tugal, Derin; Lynch, Melanie; Hujer, Andrea M.; Rudin, Susan; Perez, Federico

    2015-01-01

    Abstract Background: Klebsiella pneumoniae is an important cause of nosocomial infections, but its role in severe acute pancreatitis (SAP) is not well defined. Few cases of K. pneumoniae associated SAP have been reported. Due to the emergence of extended-spectrum beta-lactamases (ESBLs) and carbapenemases, treatment of multidrug-resistant (MDR) K. pneumoniae presents a challenge. Tigecycline and colistin have gained recent attention for their broad-spectrum antimicrobial activity. Methods: We describe a case of SAP due to K. pneumoniae bearing K. pneumoniae carbapenemase (KPC) treated successfully with colistin plus tigecycline and offer a review of similar experiences published in the literature. Results: The case reported herein required surgical drainage of multiple pancreatic abscesses and treatment with tigecycline and colistin. Our comparative analysis revealed a number of unique features associated with SAP due to K. pneumoniae: 1) underlying pancreatic injury, 2) multiple drug resistance determinants and virulence factors that complicate treatment, and 3) surgical debridement as a requirement for cure. Conclusion: As the prevalence of K. pneumoniae bearing KPC continues to increase in the healthcare setting, SAP caused by this MDR pathogen will become more common. Tigecycline plus colistin was a successful antibiotic regimen for the treatment of SAP due to K. pneumoniae bearing KPC. PMID:24850293

  5. Isolation and Characterization of Aquatic-Borne Klebsiella pneumoniae from Tropical Estuaries in Malaysia.

    PubMed

    Barati, Anis; Ghaderpour, Aziz; Chew, Li Lee; Bong, Chui Wei; Thong, Kwai Lin; Chong, Ving Ching; Chai, Lay Ching

    2016-04-15

    Klebsiella pneumoniae is an opportunistic pathogen that is responsible for causing nosocomial and community-acquired infections. Despite its common presence in soil and aquatic environments, the virulence potential of K. pneumoniae isolates of environmental origin is largely unknown. Hence, in this study, K. pneumoniae isolated from the estuarine waters and sediments of the Matang mangrove estuary were screened for potential virulence characteristics: antibiotic susceptibility, morphotype on Congo red agar, biofilm formation, presence of exopolysaccharide and capsule, possession of virulence genes (fimH, magA, ugE, wabG and rmpA) and their genomic fingerprints. A total of 55 strains of K. pneumoniae were isolated from both human-distributed sites (located along Sangga Besar River) and control sites (located along Selinsing River) where less human activity was observed, indicated that K. pneumoniae is ubiquitous in the environment. However, the detection of potentially virulent strains at the downstream of Kuala Sepetang village has suggested an anthropogenic contamination source. In conclusion, the findings from this study indicate that the Matang mangrove estuary could harbor potentially pathogenic K. pneumoniae with risk to public health. More studies are required to compare the environmental K. pneumoniae strains with the community-acquired K. pneumoniae strains.

  6. Evolution and Epidemiology of Multidrug-Resistant Klebsiella pneumoniae in the United Kingdom and Ireland

    PubMed Central

    Moradigaravand, Danesh; Martin, Veronique; Peacock, Sharon J.

    2017-01-01

    ABSTRACT Klebsiella pneumoniae is a human commensal and opportunistic pathogen that has become a leading causative agent of hospital-based infections over the past few decades. The emergence and global expansion of hypervirulent and multidrug-resistant (MDR) clones of K. pneumoniae have been increasingly reported in community-acquired and nosocomial infections. Despite this, the population genomics and epidemiology of MDR K. pneumoniae at the national level are still poorly understood. To obtain insights into these, we analyzed a systematic large-scale collection of invasive MDR K. pneumoniae isolates from hospitals across the United Kingdom and Ireland. Using whole-genome phylogenetic analysis, we placed these in the context of previously sequenced K. pneumoniae populations from geographically diverse countries and identified their virulence and drug resistance determinants. Our results demonstrate that United Kingdom and Ireland MDR isolates are a highly diverse population drawn from across the global phylogenetic tree of K. pneumoniae and represent multiple recent international introductions that are mainly from Europe but in some cases from more distant countries. In addition, we identified novel genetic determinants underlying resistance to beta-lactams, gentamicin, ciprofloxacin, and tetracyclines, indicating that both increased virulence and resistance have emerged independently multiple times throughout the population. Our data show that MDR K. pneumoniae isolates in the United Kingdom and Ireland have multiple distinct origins and appear to be part of a globally circulating K. pneumoniae population. PMID:28223459

  7. Isolation and Characterization of Aquatic-Borne Klebsiella pneumoniae from Tropical Estuaries in Malaysia

    PubMed Central

    Barati, Anis; Ghaderpour, Aziz; Chew, Li Lee; Bong, Chui Wei; Thong, Kwai Lin; Chong, Ving Ching; Chai, Lay Ching

    2016-01-01

    Klebsiella pneumoniae is an opportunistic pathogen that is responsible for causing nosocomial and community-acquired infections. Despite its common presence in soil and aquatic environments, the virulence potential of K. pneumoniae isolates of environmental origin is largely unknown. Hence, in this study, K. pneumoniae isolated from the estuarine waters and sediments of the Matang mangrove estuary were screened for potential virulence characteristics: antibiotic susceptibility, morphotype on Congo red agar, biofilm formation, presence of exopolysaccharide and capsule, possession of virulence genes (fimH, magA, ugE, wabG and rmpA) and their genomic fingerprints. A total of 55 strains of K. pneumoniae were isolated from both human-distributed sites (located along Sangga Besar River) and control sites (located along Selinsing River) where less human activity was observed, indicated that K. pneumoniae is ubiquitous in the environment. However, the detection of potentially virulent strains at the downstream of Kuala Sepetang village has suggested an anthropogenic contamination source. In conclusion, the findings from this study indicate that the Matang mangrove estuary could harbor potentially pathogenic K. pneumoniae with risk to public health. More studies are required to compare the environmental K. pneumoniae strains with the community-acquired K. pneumoniae strains. PMID:27092516

  8. Resistance of hypervirulent Klebsiella pneumoniae to both intracellular and extracellular killing of neutrophils

    PubMed Central

    Wu, Hua; Ma, Yanning

    2017-01-01

    Hypervirulent Klebsiella pneumoniae (HvKP) is hypermucoviscous organism, carrying genes of rmpA and aerobactin, causing serious community-acquired infection and metastatically spread in young healthy hosts. Neutrophils play an important role during innate immune response against bacterial infection by phagocytosis and neutrophil extracellular traps (NETs). Whether neutrophils can effectively defend against HvKP remains unclear. In this study, we observed that the HvKP was significantly more resistant to neutrophil-mediated phagocytosis and intracellular killing than classic Klebsiella pneumoniae (cKP) isolates. Although both HvKP and cKP induced NETs under scanning electron microscopy and confocal microscopy, more cKP than HvKP were trapped in NETs, and the killing by intracellular and extracellular mechanisms of neutrophils was detected only on cKP. Together, our results demonstrated that HvKP resisted to both intracellular and extracellular killing of neutrophils. PMID:28282434

  9. Klebsiella pneumonia: An unusual cause of ophthalmia neonatorum in a healthy newborn.

    PubMed

    Kumar, Jaya B; Silverstein, Evan; Wallace, David K

    2015-12-01

    Ophthalmia neonatorum is one of the most common infections during the neonatal period. Chlamydia trachomatis and Neisseria gonorrhoea must be ruled out, given their high virulence and systemic complications. We describe a case of ophthalmia neonatroum from Klebsiella pneumonia. Gram-negative organisms have been reported in hospital-acquired conjunctivitis (HAC), but we are unaware of any published reports of K. pneumonia conjunctivitis in an otherwise healthy full-term infant born in the United States who has received prophylaxis. It is important to promptly identify and treat Klebsiella conjunctivitis because it can lead to severe complications. Copyright © 2015 American Association for Pediatric Ophthalmology and Strabismus. Published by Elsevier Inc. All rights reserved.

  10. Partial excision of blaKPC from Tn4401 in carbapenem-resistant Klebsiella pneumoniae.

    PubMed

    Chen, Liang; Chavda, Kalyan D; Mediavilla, José R; Jacobs, Michael R; Levi, Michael H; Bonomo, Robert A; Kreiswirth, Barry N

    2012-03-01

    We describe a novel Tn4401 variant (Tn4401d) in epidemic Klebsiella pneumoniae clone ST258, from which a partial bla(KPC) fragment has been excised along with ISKpn7 and a partial tnpA fragment. Nested-PCR experiments confirmed that this region can be removed from distinct Tn4401 isoforms in both K. pneumoniae and Escherichia coli. This study highlights that the region surrounding bla(KPC) is undergoing recombination and that Tn4401 itself is heterogeneous and highly plastic.

  11. Emergence of KPC-Possessing Klebsiella pneumoniae in Brooklyn, New York: Epidemiology and Recommendations for Detection

    PubMed Central

    Bratu, Simona; Mooty, Mohamad; Nichani, Satyen; Landman, David; Gullans, Carl; Pettinato, Barbara; Karumudi, Usha; Tolaney, Pooja; Quale, John

    2005-01-01

    Among 257 isolates of Klebsiella pneumoniae collected in Brooklyn, NY, 24% were found to possess blaKPC. Clinical microbiology laboratories that used automated broth microdilution systems reported 15% of the KPC-possessing isolates as susceptible to imipenem. The imipenem MIC was found to be markedly affected by the inoculum. For accurate detection of KPC-possessing K. pneumoniae, particular attention should be paid to proper inoculum preparation for broth-based susceptibility methods. In addition, using ertapenem or meropenem for class reporting of carbapenem susceptibility will improve detection. PMID:15980389

  12. Outbreak of OXA-48-Positive Carbapenem-Resistant Klebsiella pneumoniae Isolates in France▿

    PubMed Central

    Cuzon, Gaelle; Ouanich, Jocelyne; Gondret, Remy; Naas, Thierry; Nordmann, Patrice

    2011-01-01

    Seventeen Klebsiella pneumoniae isolates producing the OXA-48 carbapenemase, obtained from 10 patients hospitalized from April to June 2010, mostly in the medical intensive care unit of the Villeneuve-Saint-Georges Hospital in a suburb of Paris, France, were analyzed. Seven patients were infected, of whom five were treated at least with a carbapenem, and five patients died. Molecular analysis showed that the isolates belonged to a single clone that harbored a 70-kb plasmid carrying the blaOXA-48 gene and coproduced CTX-M-15 and TEM-1 β-lactamases. This is the first reported outbreak of OXA-48-producing K. pneumoniae isolates in France. PMID:21343451

  13. Production of 1,3-propanediol by Klebsiella pneumoniae from glycerol broth.

    PubMed

    Cheng, Ke-Ke; Zhang, Jian-An; Liu, De-Hua; Sun, Yan; Yang, Ming-De; Xu, Jing-Ming

    2006-11-01

    Broth containing 152 g glycerol l(-1) from Candida krusei culture was converted to 1,3-propanediol by Klebsiella pneumoniae. Residual glucose in the broth promoted growth of K. pneumoniae while acetate was inhibitory. After desalination treatment of glycerol broth by electrodialysis, the acetate in the broth was removed. A fed-batch culture with electrodialytically pretreated broth as substrate was developed giving 53 g 1,3-propanediol l(-1) with a yield of 0.41 g g(-1) glycerol and a productivity of 0.94 g l(-1) h(-1).

  14. Genetic profiling of Klebsiella pneumoniae: comparison of pulsed field gel electrophoresis and random amplified polymorphic DNA

    PubMed Central

    Ashayeri-Panah, Mitra; Eftekhar, Fereshteh; Ghamsari, Maryam Mobarak; Parvin, Mahmood; Feizabadi, Mohammad Mehdi

    2013-01-01

    In this study, the discriminatory power of pulsed field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD) methods for subtyping of 54 clinical isolates of Klebsiella pneumoniae were compared. All isolates were typeable by RAPD, while 3.6% of them were not typeable by PFGE. The repeatability of both typing methods were 100% with satisfying reproducibility (≥ 95%). Although the discriminatory power of PFGE was greater than RAPD, both methods showed sufficient discriminatory power (DI > 0.95) which reflects the heterogeneity among the K. pneumoniae isolates. An optimized RAPD protocol is less technically demanding and time consuming that makes it a reliable typing method and competitive with PFGE. PMID:24516423

  15. Population structure of KPC-producing Klebsiella pneumoniae isolates from midwestern U.S. hospitals.

    PubMed

    Wright, Meredith S; Perez, Federico; Brinkac, Lauren; Jacobs, Michael R; Kaye, Keith; Cober, Eric; van Duin, David; Marshall, Steven H; Hujer, Andrea M; Rudin, Susan D; Hujer, Kristine M; Bonomo, Robert A; Adams, Mark D

    2014-08-01

    Genome sequencing of carbapenem-resistant Klebsiella pneumoniae isolates from regional U.S. hospitals was used to characterize strain diversity and the bla(KPC) genetic context. A phylogeny based on core single-nucleotide variants (SNVs) supports a division of sequence type 258 (ST258) into two distinct groups. The primary differences between the groups are in the capsular polysaccharide locus (cps) and their plasmid contents. A strict association between clade and KPC variant was found. The bla(KPC) gene was found on variants of two plasmid backbones. This study indicates that highly similar K. pneumoniae subpopulations coexist within the same hospitals over time.

  16. Emergence of two distinct subpopulations from Klebsiella pneumoniae grown in the stimulated microgravity environment.

    PubMed

    Wang, Haili; Li, Wenliang; Gu, Lixiao; Gao, Xiaofang; Ni, Bin; Deng, Haiteng; Yang, Ruifu; Han, Yanping

    2017-09-01

    To isolate and characterize the two phenotypically distinct subpopulations from Klebsiella pneumoniae clonal cultures grown in the simulate microgravity environment. Here clonal culture of K. pneumoniae strain ATCC BAA-1705 was grown within a vertically rotating wall vessel bioreactor. Microscopic, colony staining, biofilm assays and quantitative proteomics were used to define the features of subpopulations. Two subpopulations were isolated based on colony appearance and bacterial morphology and indicated the different capability of biofilm formation and antibiotics resistance. These findings would raise a possibility of understanding the adaptive roles of bacterial subpopulations formed under certain conditions from the viewpoint of population variation.

  17. Isolation of KPC 3-producing Enterobacter aerogenes in a patient colonized by MDR Klebsiella pneumoniae.

    PubMed

    Venditti, Carolina; Villa, Laura; Capone, Alessandro; Fortini, Daniela; D'Arezzo, Silvia; Nisii, Carla; Bordi, Eugenio; Puro, Vincenzo; Antonini, Mario; Carattoli, Alessandra; Cataldo, Maria Adriana; Petrosillo, Nicola; Di Caro, Antonino

    2016-10-01

    We describe the interspecies transmission of the plasmid-mediated blaKPC-3 gene, which confers carbapenem resistance, between clinically relevant gram-negative bacteria in a single patient. A KPC-3 producing Enterobacter aerogenes was isolated from a hospitalized patient previously colonized and then infected by a Klebsiella pneumoniae ST101 carrying the blaKPC-3 gene. The strains showed identical plasmids. Since intense horizontal exchanges among bacteria can occur in the gut, clinicians should be aware that patients colonized by carbapenem-resistant K. pneumoniae could become carriers of other carbapenem-resistant Enterobacteriaceae.

  18. Population Structure of KPC-Producing Klebsiella pneumoniae Isolates from Midwestern U.S. Hospitals

    PubMed Central

    Wright, Meredith S.; Perez, Federico; Brinkac, Lauren; Jacobs, Michael R.; Kaye, Keith; Cober, Eric; van Duin, David; Marshall, Steven H.; Hujer, Andrea M.; Rudin, Susan D.; Hujer, Kristine M.

    2014-01-01

    Genome sequencing of carbapenem-resistant Klebsiella pneumoniae isolates from regional U.S. hospitals was used to characterize strain diversity and the blaKPC genetic context. A phylogeny based on core single-nucleotide variants (SNVs) supports a division of sequence type 258 (ST258) into two distinct groups. The primary differences between the groups are in the capsular polysaccharide locus (cps) and their plasmid contents. A strict association between clade and KPC variant was found. The blaKPC gene was found on variants of two plasmid backbones. This study indicates that highly similar K. pneumoniae subpopulations coexist within the same hospitals over time. PMID:24913165

  19. KPC-2-producing Klebsiella pneumoniae in a hospital in the Midwest region of Brazil.

    PubMed

    Biberg, Camila Arguelo; Rodrigues, Ana Claudia Souza; do Carmo, Sidiane Ferreira; Chaves, Claudia Elizabeth Volpe; Gales, Ana Cristina; Chang, Marilene Rodrigues

    2015-06-01

    The emergence of β-lactamase-producing Enterobacteriaceae in the last few decades has become major challenge faced by hospitals. In this study, isolates of Klebsiella pneumoniae carbapenemase-2 (KPC-2)-producing K. pneumoniae from a tertiary hospital in Mato Grosso do Sul, Brazil, were characterized. Bacterial identification was performed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF; Bruker Daltonics, Germany) mass spectrometry. The minimum inhibitory concentrations of carbapenems were determined using the agar dilution method as recommended by the Clinical Laboratory Standards Institute guidelines. Carbapenemase production was detected using the modified Hodge test (MHT) and polymerase chain reaction (PCR), followed by DNA sequencing. Of 360 (12.2%) K. pneumoniae isolates obtained between May 2009 and May 2010, 44 (12.2%) were carbapenem nonsusceptible. Of these 44 isolates, thirty-six K. pneumoniae isolates that were positive by MHT and PCR carried the bla KPC-2 gene. Thus, KPC-2producing Klebsiella pneumoniae has been present in a Brazilian hospital located in the Midwest region since at least 2009.

  20. KPC-2-producing Klebsiella pneumoniae in a hospital in the Midwest region of Brazil

    PubMed Central

    Biberg, Camila Arguelo; Rodrigues, Ana Claudia Souza; do Carmo, Sidiane Ferreira; Chaves, Claudia Elizabeth Volpe; Gales, Ana Cristina; Chang, Marilene Rodrigues

    2015-01-01

    The emergence of β-lactamase-producing Enterobacteriaceae in the last few decades has become major challenge faced by hospitals. In this study, isolates of Klebsiella pneumoniae carbapenemase-2 (KPC-2)-producing K. pneumoniae from a tertiary hospital in Mato Grosso do Sul, Brazil, were characterized. Bacterial identification was performed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF; Bruker Daltonics, Germany) mass spectrometry. The minimum inhibitory concentrations of carbapenems were determined using the agar dilution method as recommended by the Clinical Laboratory Standards Institute guidelines. Carbapenemase production was detected using the modified Hodge test (MHT) and polymerase chain reaction (PCR), followed by DNA sequencing. Of 360 (12.2%) K. pneumoniae isolates obtained between May 2009 and May 2010, 44 (12.2%) were carbapenem nonsusceptible. Of these 44 isolates, thirty-six K. pneumoniae isolates that were positive by MHT and PCR carried the bla KPC-2 gene. Thus, KPC-2producing Klebsiella pneumoniae has been present in a Brazilian hospital located in the Midwest region since at least 2009. PMID:26273265

  1. Evaluation of the efficacy of a bacteriophage in the treatment of pneumonia induced by multidrug resistance Klebsiella pneumoniae in mice.

    PubMed

    Cao, Fang; Wang, Xitao; Wang, Linhui; Li, Zhen; Che, Jian; Wang, Lili; Li, Xiaoyu; Cao, Zhenhui; Zhang, Jiancheng; Jin, Liji; Xu, Yongping

    2015-01-01

    Multidrug-resistant Klebsiella pneumoniae (MRKP) has steadily grown beyond antibiotic control. However, a bacteriophage is considered to be a potential antibiotic alternative for treating bacterial infections. In this study, a lytic bacteriophage, phage 1513, was isolated using a clinical MRKP isolate KP 1513 as the host and was characterized. It produced a clear plaque with a halo and was classified as Siphoviridae. It had a short latent period of 30 min, a burst size of 264 and could inhibit KP 1513 growth in vitro with a dose-dependent pattern. Intranasal administration of a single dose of 2×10(9) PFU/mouse 2 h after KP 1513 inoculation was able to protect mice against lethal pneumonia. In a sublethal pneumonia model, phage-treated mice exhibited a lower level of K. pneumoniae burden in the lungs as compared to the untreated control. These mice lost less body weight and exhibited lower levels of inflammatory cytokines in their lungs. Lung lesion conditions were obviously improved by phage therapy. Therefore, phage 1513 has a great effect in vitro and in vivo, which has potential to be used as an alternative to an antibiotic treatment of pneumonia that is caused by the multidrug-resistant K. pneumoniae.

  2. Phenotypic detection of Klebsiella pneumoniae carbapenemase among burns patients: first report from Iran.

    PubMed

    Rastegar Lari, Abdolaziz; Azimi, Leila; Rahbar, Mohammad; Fallah, Fatemeh; Alaghehbandan, Reza

    2013-02-01

    Resistance to antimicrobial agents such as carbapenems among enterobacteriacea has been increasing, especially in Klebsiella pneumonia that produces variety of enzymes including Klebsiella pneumoniae carbapenemase (KPC). This study is the first report of its kind investigating the resistance to carbapenems among burns patients in Iran. During a 6-month period, 28 hospitalized burn patients who required to be placed on broad spectrum antibiotics were studied. Isolated species identified by routine biochemical test. Susceptibility testing for these species was performed by recommended the CLSI guidelines method. The tested antibiotics included cefotaxime, cefepime, aztreonam, imipenem, amoxicillin+clavulonic acid, gentamicin, amikacin, tobramycin, tetracycline, and trimethoprim-sulfamethoxazole, and chloramphenicol. For determination of KPC in phenotypical forms, Modified Hodge Test was utilized as per CLSI recommendation. Thirty-five Klebsiella spp. were isolated from 28 hospitalized patients. Nineteen out of 35 Klebsiella isolates were resistant to imipenem and that all of them had positive KPC. Nine of imipenem resistant isolates were also resistant to all tested antibiotics. Mortality rate among patients with positive KPC was 33%. High rate of multi-drug resistant (MDR) strains in isolates with positive KPC is a major challenge in Iran and that it could cause an increase in both mortality and morbidity among burn patients. Thus, appropriate infection control measures and guidelines are needed to prevent such infections among burn patients. Copyright © 2012 Elsevier Ltd and ISBI. All rights reserved.

  3. Genome-Wide Identification of Klebsiella pneumoniae Fitness Genes during Lung Infection.

    PubMed

    Bachman, Michael A; Breen, Paul; Deornellas, Valerie; Mu, Qiao; Zhao, Lili; Wu, Weisheng; Cavalcoli, James D; Mobley, Harry L T

    2015-06-09

    Klebsiella pneumoniae is an urgent public health threat because of resistance to carbapenems, antibiotics of last resort against Gram-negative bacterial infections. Despite the fact that K. pneumoniae is a leading cause of pneumonia in hospitalized patients, the bacterial factors required to cause disease are poorly understood. Insertion site sequencing combines transposon mutagenesis with high-throughput sequencing to simultaneously screen thousands of insertion mutants for fitness defects during infection. Using the recently sequenced K. pneumoniae strain KPPR1 in a well-established mouse model of pneumonia, insertion site sequencing was performed on a pool of >25,000 transposon mutants. The relative fitness requirement of each gene was ranked based on the ratio of lung to inoculum read counts and concordance between insertions in the same gene. This analysis revealed over 300 mutants with at least a 2-fold fitness defect and 69 with defects ranging from 10- to >2,000-fold. Construction of 6 isogenic mutants for use in competitive infections with the wild type confirmed their requirement for lung fitness. Critical fitness genes included those for the synthesis of branched-chain and aromatic amino acids that are essential in mice and humans, the transcriptional elongation factor RfaH, and the copper efflux pump CopA. The majority of fitness genes were conserved among reference strains representative of diverse pathotypes. These results indicate that regulation of outer membrane components and synthesis of amino acids that are essential to its host are critical for K. pneumoniae fitness in the lung. Klebsiella pneumoniae is a bacterium that commonly causes pneumonia in patients after they are admitted to the hospital. K. pneumoniae is becoming resistant to all available antibiotics, and when these infections spread to the bloodstream, over half of patients die. Since currently available antibiotics are failing, we must discover new ways to treat these infections

  4. Spread of OXA-48-Positive Carbapenem-Resistant Klebsiella pneumoniae Isolates in Istanbul, Turkey▿

    PubMed Central

    Carrër, Amélie; Poirel, Laurent; Eraksoy, Haluk; Cagatay, A. Atahan; Badur, Selim; Nordmann, Patrice

    2008-01-01

    The first outbreak of carbapenem-resistant Klebsiella pneumoniae isolates producing the plasmid-encoded carbapenem-hydrolyzing oxacillinase OXA-48 is reported. The 39 isolates belonged to two different clones and were collected at the University Hospital of Istanbul, Turkey, from May 2006 to February 2007, and they coproduced various β-lactamases (SHV-12, OXA-9, and TEM-1 for clone A and CTX-M-15, TEM-1, and OXA-1 for clone B). PMID:18519712

  5. Liver abscess due to Klebsiella pneumoniae in a healthy 12-year-old boy

    PubMed Central

    Yoon, Da Hye; Jeon, Yeon Jin; Bae, E Young; Jeong, Dae Chul

    2013-01-01

    Pyogenic liver abscess (PLA) is rare in healthy children. We report a case of PLA in an immunocompetent 12-year-old boy. Percutaneous catheter drainage was performed for the abscess. In addition, parenteral antibiotics were administered for 3 weeks. Klebsiella pneumoniae was detected in the culture of blood and drained fluid. Here, we present this case and a brief review of the literature on this subject. PMID:24348663

  6. Environmental persistence of OXA-48-producing Klebsiella pneumoniae in a French intensive care unit.

    PubMed

    Pantel, Alix; Richaud-Morel, Brigitte; Cazaban, Michel; Bouziges, Nicole; Sotto, Albert; Lavigne, Jean-Philippe

    2016-03-01

    The spread of carbapenemase-producing Gram-negative rods is an emerging global problem. This study describes the epidemiologic features of an outbreak caused by an environmental reservoir of OXA-48-producing Klebsiella pneumoniae caused by persistence of the bacteria during 20 months in an intensive care unit in France. This report emphasizes the importance of early environmental screening to interrupt the transmission of carbapenemase-producingEnterobacteriaceae.

  7. A simple method for extracting C-phycocyanin from Spirulina platensis using Klebsiella pneumoniae.

    PubMed

    Zhu, Y; Chen, X B; Wang, K B; Li, Y X; Bai, K Z; Kuang, T Y; Ji, H B

    2007-02-01

    C-phycocyanin (C-PC) was extracted from fresh Spirulina platensis by deploying a species of non-pathogenic nitrogen-fixing bacteria, namely, Klebsiella pneumoniae. The algal slurry was neither washed nor centrifuged; the bacterial culture was poured into the slurry, the vessel sealed, and crude C-PC extracted after about 24 h. The extraction was clean and efficient, and the purity and concentration of C-PC proved to be of adequate quality.

  8. Catalytic Buffering: Development of the Fluoride-Resistant Ureases of Klebsiella pneumoniae

    DTIC Science & Technology

    2005-10-01

    for nickel incorporation into E . coli to produce active urease in the clones. E . coli in vitro mutagenesis was followed by screening of the lysed...urease of the gram-negative enteric bacteria Klebsiella pneumoniae (formerly aerogenes ) was chosen for these studies. The KAU urease is cloned...mutagenesis, creates random DNA point mutations using an E . coli mutator strain. The FR mutant plasmids isolated from this treatment can then be

  9. Oxygen sensitivity of the nifLA promoter of Klebsiella pneumoniae

    SciTech Connect

    Kong, Q.T.; Wu, Q.L.; Ma, Z.F.; Shen, S.C.

    1986-05-01

    Oxygen sensitivity of the nifLA promoter of Klebsiella pneumoniae has been demonstrated. Studies on the oxygen regulation of nifB-lacZ and nifH-lacZ fusions in the presence of the nifLA operon, which contains either an intact or a deleted nifL gene, indicate that possible both the nifL promoter and the nifL product are responsible for nif repression by oxygen.

  10. Oxygen sensitivity of the nifLA promoter of Klebsiella pneumoniae.

    PubMed Central

    Kong, Q T; Wu, Q L; Ma, Z F; Shen, S C

    1986-01-01

    Oxygen sensitivity of the nifLA promoter of Klebsiella pneumoniae has been demonstrated. Studies on the oxygen regulation of nifB-lacZ and nifH-lacZ fusions in the presence of the nifLA operon, which contains either an intact or a deleted nifL gene, indicate that possibly both the nifL promoter and the nifL product are responsible for nif repression by oxygen. PMID:3082858

  11. Mother-To-Child Transmission of KPC Carbapenemase-Producing Klebsiella Pneumoniae at Birth.

    PubMed

    Bonfanti, Paolo; Bellù, Roberto; Principe, Luigi; Caramma, Ilaria; Condò, Manuela; Giani, Tommaso; Rossolini, Gian Maria; Luzzaro, Francesco

    2017-02-01

    We report on a mother-to-child transmission of KPC carbapenemase-producing Klebsiella pneumoniae at birth followed by subsequent cases in the neonatal intensive care unit. Molecular analysis of isolates showed production of KPC-3 enzyme. The only potential risk factor identified for the mother was previous activity as a caregiver. Present findings suggest consideration of proactive surveillance in pregnant women with risk factors for colonization.

  12. Community-Acquired Pyelonephritis in Pregnancy Caused by KPC-Producing Klebsiella pneumoniae

    PubMed Central

    Khatri, Asma; Naeger Murphy, Nina; Wiest, Peter; Osborn, Melissa; Garber, Kathleen; Hecker, Michelle; Hurless, Kelly; Rudin, Susan D.; Jacobs, Michael R.; Kalayjian, Robert C.; Salata, Robert A.; van Duin, David; Harris, Patrick N. A.

    2015-01-01

    Carbapenem-resistant Enterobacteriaceae (CRE) usually infect patients with significant comorbidities and health care exposures. We present a case of a pregnant woman who developed community-acquired pyelonephritis caused by KPC-producing Klebsiella pneumoniae. Despite antibiotic treatment, she experienced spontaneous prolonged rupture of membranes, with eventual delivery of a healthy infant. This report demonstrates the challenge that CRE may pose to the effective treatment of common infections in obstetric patients, with potentially harmful consequences to maternal and neonatal health. PMID:26185273

  13. Novel VIM Metallo-β-Lactamase Variant, VIM-24, from a Klebsiella pneumoniae Isolate from Colombia▿

    PubMed Central

    Montealegre, Maria Camila; Correa, Adriana; Briceño, David F.; Rosas, Natalia C.; De La Cadena, Elsa; Ruiz, Sory J.; Mojica, Maria F.; Camargo, Ruben Dario; Zuluaga, Ivan; Marin, Adriana; Quinn, John P.; Villegas, Maria Virginia

    2011-01-01

    We report the emergence of a novel VIM variant (VIM-24) in a Klebsiella pneumoniae isolate in Colombia. The isolate displays MICs for carbapenems below the resistance breakpoints, posing a real challenge for its detection. The blaVIM-24 gene was located within a class 1 integron carried on a large plasmid. Further studies are needed to clarify its epidemiological and clinical impact. PMID:21282438

  14. Thermotolerant non-fecal source Klebsiella pneumoniae: validity of the fecal coliform test in recreational waters.

    PubMed Central

    Caplenas, N R; Kanarek, M S

    1984-01-01

    Wisconsin pulp and paper mill processing plants were evaluated for fecal coliform and total Klebsiella (i.e., thermotolerant and thermointolerant) bacterial concentrations. Using the standard fecal coliform test, up to 90 per cent of non-fecal source thermotolerant K. pneumoniae was falsely identified as fecal source bacteria. Since there is a lack of specificity in the currently used standard for fecal coliform evaluation, a more reliable health risk assessment for fecal coliform bacteria is recommended. PMID:6388365

  15. Effect of Molybdenum Starvation and Tungsten on the Synthesis of Nitrogenase Components in Klebsiella pneumoniae

    PubMed Central

    Brill, Winston J.; Steiner, Ann L.; Shah, Vinod K.

    1974-01-01

    Klebsiella pneumoniae M5a1 grows well in the presence or absence of molybdenum in media containing excess NH4+. However, growth on N2 is completely dependent on the presence of molybdenum in the medium. Tungstate competes with the molybdate requirement during growth on N2. In molybdenum-depleted medium, neither protein component of nitrogenase is active and neither component can be detected antigenically. These data provide evidence that molybdenum is an inducer of nitrogenase synthesis. PMID:4598014

  16. An outbreak of colistin-resistant Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae in the Netherlands (July to December 2013), with inter-institutional spread.

    PubMed

    Weterings, V; Zhou, K; Rossen, J W; van Stenis, D; Thewessen, E; Kluytmans, J; Veenemans, J

    2015-08-01

    We describe an outbreak of Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae (KPC-KP) ST258 that occurred in two institutions (a hospital and a nursing home) in the Netherlands between July and December 2013. In total, six patients were found to be positive for KPC-KP. All isolates were resistant to colistin and exhibited reduced susceptibility to gentamicin and tigecycline. In all settings, extensive environmental contamination was found. Whole genome sequencing revealed the presence of bla KPC-2 and bla SHV-12 genes, as well as the close relatedness of patient and environmental isolates. In the hospital setting, one transmission was detected, despite contact precautions. After upgrading to strict isolation, no further spread was found. After the transfer of the index patient to a nursing home in the same region, four further transmissions occurred. The outbreak in the nursing home was controlled by transferring all KPC-KP-positive residents to a separate location outside the nursing home, where a dedicated nursing team cared for patients. This outbreak illustrates that the spread of pan-resistant Enterobacteriaceae can be controlled, but may be difficult, particularly in long-term care facilities. It, therefore, poses a major threat to patient safety. Clear guidelines to control reservoirs in and outside the hospitals are urgently needed.

  17. First evidence of polar flagella in Klebsiella pneumoniae isolated from a patient with neonatal sepsis.

    PubMed

    Carabarin-Lima, Alejandro; León-Izurieta, Libia; Rocha-Gracia, Rosa Del Carmen; Castañeda-Lucio, Miguel; Torres, Carmen; Gutiérrez-Cazarez, Zita; González-Posos, Sirenia; Martínez de la Peña, Claudia F; Martinez-Laguna, Ygnacio; Lozano-Zarain, Patricia

    2016-08-01

    The genus Klebsiella belongs to the family Enterobacteriaceae, and is currently considered to be non-motile and non-flagellated. In the present work, 25 Klebsiella strains isolated from nosocomial infections were assessed for motility under different growth conditions. One Klebsiella isolate, KpBUAP021, demonstrated a swim-like motility phenotype. The K. pneumoniae genotype was confirmed by 16S rRNA and rpoB gene sequence analysis. Multilocus sequence typing analysis also revealed that the KpBUAP021 strain places it in the ST345 sequence type, and belongs to the phylogenetic Kpl group. Transmission electron microscopy and the Ryu staining technique revealed that KpBUAP021 expresses polar flagella. Finally, the presence of fliC, fliA and flgH genes in this K. pneumoniae strain was confirmed. This report presents the first evidence for flagella-mediated motility in a K. pneumoniae clinical isolate, and represents an important finding related to its evolution and pathogenic potential.

  18. Molecular typing and virulence analysis of multidrug resistant Klebsiella pneumoniae clinical isolates recovered from Egyptian hospitals

    PubMed Central

    Wasfi, Reham; Elkhatib, Walid F.; Ashour, Hossam M.

    2016-01-01

    Klebsiella pneumonia infection rates have increased dramatically. Molecular typing and virulence analysis are powerful tools that can shed light on Klebsiella pneumonia infections. Whereas 77.7% (28/36) of clinical isolates indicated multidrug resistant (MDR) patterns, 50% (18/36) indicated carpabenem resistance. Gene prevalence for the AcrAB efflux pump (82.14%) was more than that of the mdtK efflux pump (32.14%) in the MDR isolates. FimH-1 and mrkD genes were prevalent in wound and blood isolates. FimH-1 gene was prevalent in sputum while mrkD gene was prevalent in urine. Serum resistance associated with outer membrane protein coding gene (traT) was found in all blood isolates. IucC, entB, and Irp-1 were detected in 32.14%, 78.5% and 10.7% of MDR isolates, respectively. We used two Polymerase Chain Reaction (PCR) analyses: Enterobacterial Repetitive Intergenic Consensus (ERIC) and Random Amplified Polymorphic DNA (RAPD). ERIC-PCR revealed 21 and RAPD-PCR revealed 18 distinct patterns of isolates with similarity ≥80%. ERIC genotyping significantly correlated with resistance patterns and virulence determinants. RAPD genotyping significantly correlated with resistance patterns but not with virulence determinants. Both RAPD and ERIC genotyping methods had no correlation with the capsule types. These findings can help up better predict MDR Klebsiella pneumoniae outbreaks associated with specific genotyping patterns. PMID:28004732

  19. Complete Genome Sequence of Klebsiella pneumoniae subsp. pneumoniae KP617, Coproducing OXA-232 and NDM-1 Carbapenemases, Isolated in South Korea

    PubMed Central

    Kwon, Taesoo; Yang, Ji Woo; Lee, Sanghyun; Yun, Mi-ran; Yoo, Won Gi; Kim, Hwa Su

    2016-01-01

    The prevalence of Klebsiella pneumoniae coproducing carbapenemase metallo-β-lactamase 1 (NDM-1) and OXA-48 has been increasing globally since 2013. The complete genome of KP617 was sequenced and assembled into a circular chromosome and two plasmids. This sequence provides the genetic background for understanding the evolution of carbapenemase genes in K. pneumoniae KP617. PMID:26769936

  20. Demonstration that a Klebsiella pneumoniae subsp. pneumoniae isolated from an insect (Nezara viridula) harbors a functional plasmid-borne type IV secretion system

    USDA-ARS?s Scientific Manuscript database

    Previously, we reported the isolation of Klebsiella pneumoniae subspecies pneumoniae strain Kp 5-1 from a southern green stink bug (Nezara viridula) that is a significant pest of numerous economically important crops. We subsequently sequenced the strains whole genome. Here, we report the presence...

  1. Plasmid Dynamics in KPC-Positive Klebsiella pneumoniae during Long-Term Patient Colonization.

    PubMed

    Conlan, Sean; Park, Morgan; Deming, Clayton; Thomas, Pamela J; Young, Alice C; Coleman, Holly; Sison, Christina; Weingarten, Rebecca A; Lau, Anna F; Dekker, John P; Palmore, Tara N; Frank, Karen M; Segre, Julia A

    2016-06-28

    Carbapenem-resistant Klebsiella pneumoniae strains are formidable hospital pathogens that pose a serious threat to patients around the globe due to a rising incidence in health care facilities, high mortality rates associated with infection, and potential to spread antibiotic resistance to other bacterial species, such as Escherichia coli Over 6 months in 2011, 17 patients at the National Institutes of Health (NIH) Clinical Center became colonized with a highly virulent, transmissible carbapenem-resistant strain of K. pneumoniae Our real-time genomic sequencing tracked patient-to-patient routes of transmission and informed epidemiologists' actions to monitor and control this outbreak. Two of these patients remained colonized with carbapenemase-producing organisms for at least 2 to 4 years, providing the opportunity to undertake a focused genomic study of long-term colonization with antibiotic-resistant bacteria. Whole-genome sequencing studies shed light on the underlying complex microbial colonization, including mixed or evolving bacterial populations and gain or loss of plasmids. Isolates from NIH patient 15 showed complex plasmid rearrangements, leaving the chromosome and the blaKPC-carrying plasmid intact but rearranging the two other plasmids of this outbreak strain. NIH patient 16 has shown continuous colonization with blaKPC-positive organisms across multiple time points spanning 2011 to 2015. Genomic studies defined a complex pattern of succession and plasmid transmission across two different K. pneumoniae sequence types and an E. coli isolate. These findings demonstrate the utility of genomic methods for understanding strain succession, genome plasticity, and long-term carriage of antibiotic-resistant organisms. In 2011, the NIH Clinical Center had a nosocomial outbreak involving 19 patients who became colonized or infected with blaKPC-positive Klebsiella pneumoniae Patients who have intestinal colonization with blaKPC-positive K. pneumoniae are at

  2. Plasmid transferability of KPC into a virulent K2 serotype Klebsiella pneumoniae

    PubMed Central

    2014-01-01

    Background KPC-producing carbapenem-resistant Klebsiella pneumoniae (CRKP) infections are associated with high mortality; however, their virulence determinants are not well defined. Methods We investigated the virulence and plasmid transferability among KPC-containing K. pneumoniae isolates. Results KPC-2 and -3 were successfully conjugated and retained by a virulent K2 K. pneumoniae recipient isolate. Antimicrobial susceptibility testing showed KPC-2 and -3 donor strains were resistant to more than four classes of antibiotics while the K2 isolate was only initially resistant to ampicillin. After conjugation of KPC-2 and -3, the K2 K. pneumoniae transconjugants became resistant to all beta-lactams. Additionally, the KPC K2 K. pneumoniae transconjugants continued to retain its high serum resistance and murine lethality. Conclusions Conjugation and retainment of KPC by virulent K2 K. pneumoniae and the ability of the tranconjugants to maintain its high serum resistance and murine lethality after conjugation was demonstrated in this study. These findings are concerning for the potential of KPC-like genes to disseminate among virulent K. pneumoniae isolates. PMID:24678611

  3. Plasmid transferability of KPC into a virulent K2 serotype Klebsiella pneumoniae.

    PubMed

    Siu, Leung-Kei Kristopher; Huang, David B; Chiang, Tom

    2014-03-31

    KPC-producing carbapenem-resistant Klebsiella pneumoniae (CRKP) infections are associated with high mortality; however, their virulence determinants are not well defined. We investigated the virulence and plasmid transferability among KPC-containing K. pneumoniae isolates. KPC-2 and -3 were successfully conjugated and retained by a virulent K2 K. pneumoniae recipient isolate. Antimicrobial susceptibility testing showed KPC-2 and -3 donor strains were resistant to more than four classes of antibiotics while the K2 isolate was only initially resistant to ampicillin. After conjugation of KPC-2 and -3, the K2 K. pneumoniae transconjugants became resistant to all beta-lactams. Additionally, the KPC K2 K. pneumoniae transconjugants continued to retain its high serum resistance and murine lethality. Conjugation and retainment of KPC by virulent K2 K. pneumoniae and the ability of the tranconjugants to maintain its high serum resistance and murine lethality after conjugation was demonstrated in this study. These findings are concerning for the potential of KPC-like genes to disseminate among virulent K. pneumoniae isolates.

  4. Virulence of Klebsiella pneumoniae isolates harboring bla KPC-2 carbapenemase gene in a Caenorhabditis elegans model.

    PubMed

    Lavigne, Jean-Philippe; Cuzon, Gaelle; Combescure, Christophe; Bourg, Gisèle; Sotto, Albert; Nordmann, Patrice

    2013-01-01

    Klebsiella pneumoniae carbapenemase (KPC) is a carbapenemase increasingly reported worldwide in Enterobacteriaceae. The aim of this study was to analyze the virulence of several KPC-2-producing K. pneumoniae isolates. The studied strains were (i) five KPC-2 clinical strains from different geographical origins, belonging to different ST-types and possessing plasmids of different incompatibility groups; (ii) seven transformants obtained after electroporation of either these natural KPC plasmids or a recombinant plasmid harboring only the bla KPC-2 gene into reference strains K. pneumoniae ATCC10031/CIP53153; and (iii) five clinical strains cured of plasmids. The virulence of K. pneumoniae isolates was evaluated in the Caenorhabditis elegans model. The clinical KPC producers and transformants were significantly less virulent (LT50: 5.5 days) than K. pneumoniae reference strain (LT50: 4.3 days) (p<0.01). However, the worldwide spread KPC-2 positive K. pneumoniae ST258 strains and reference strains containing plasmids extracted from K. pneumoniae ST258 strains had a higher virulence than KPC-2 strains belonging to other ST types (LT50: 5 days vs. 6 days, p<0.01). The increased virulence observed in cured strains confirmed this trend. The bla KPC-2 gene itself was not associated to increased virulence.

  5. Genome Sequences of Multidrug-Resistant, Colistin-Susceptible and -Resistant Klebsiella pneumoniae Clinical Isolates from Pakistan

    PubMed Central

    Crawford, Matthew A.; Timme, Ruth; Lomonaco, Sara; Lascols, Christine; Fisher, Debra J.; Sharma, Shashi K.; Strain, Errol; Allard, Marc W.; Brown, Eric W.; McFarland, Melinda A.; Croley, Tim; Hammack, Thomas S.; Weigel, Linda M.; Anderson, Kevin; Hodge, David R.; Pillai, Segaran P.; Morse, Stephen A.; Khan, Erum

    2016-01-01

    The emergence and spread of colistin resistance among multidrug-resistant (MDR) Klebsiella pneumoniae represent a critical threat to global health. Here, we report the complete genome sequences of 10 MDR, colistin-susceptible and -resistant K. pneumoniae clinical isolates obtained in Pakistan between 2010 and 2013. PMID:27979956

  6. Genetic mechanisms of multidrug resistance among Klebsiella pneumoniae isolates from food-producing animals and humans in Lagos, Nigeria

    USDA-ARS?s Scientific Manuscript database

    Klebsiella pneumoniae is an opportunistic pathogen that commonly causes hospital and community acquired bacterial infections in humans. The emergence and rapid spread of multi- drug resistant (MDR) K. pneumoniae is causing drug therapy failure amid patients leading to poor antibiotic management glob...

  7. Molecular characterization of DHA-1-producing Klebsiella pneumoniae isolates collected during a 4-year period in an intensive care unit.

    PubMed

    Compain, Fabrice; Decré, Dominique; Fulgencio, Jean-Pierre; Berraho, Sfia; Arlet, Guillaume; Verdet, Charlotte

    2014-10-01

    Seventeen Klebsiella pneumoniae isolates producing DHA-1 β-lactamase were collected in an intensive care unit between 2006 and 2010. Molecular analysis revealed the predominance of ST48 and ST1263 clones of K. pneumoniae and the spread of DHA-1-encoding plasmids belonging to incompatibility group IncL/M or IncHI2.

  8. Fulminant mediastinitis due to extended-spectrum beta-lactamase-producing Klebsiella pneumoniae: atypical presentation and spreading following cardiac surgery.

    PubMed

    Valenzuela, Horacio; Carrascal, Yolanda; Maroto, Laura; Arce, Nuria

    2013-05-01

    Mediastinitis due to Klebsiella pneumoniae, related to thoracic wall contamination after cardiac surgery, has rarely been described. We aim to report a case of fulminant mediastinitis due to extended-spectrum beta-lactamase-producing K. pneumoniae, secondary to a disseminated concomitant pulmonary infection. The patient remained pauci-symptomatic until clinical manifestations of sepsis acutely appeared.

  9. Draft Genome Sequences of Klebsiella pneumoniae Clinical Type Strain ATCC 13883 and Three Multidrug-Resistant Clinical Isolates.

    PubMed

    Arivett, Brock A; Ream, David C; Fiester, Steven E; Mende, Katrin; Murray, Clinton K; Thompson, Mitchell G; Kanduru, Shrinidhi; Summers, Amy M; Roth, Amanda L; Zurawski, Daniel V; Actis, Luis A

    2015-01-15

    Klebsiella pneumoniae is a Gram-negative human pathogen capable of causing hospital-acquired infections with an increasing risk to human health. The total DNA from four clinically relevant strains was sequenced to >100× coverage, providing high-quality genome assemblies for K. pneumoniae strains ATCC 13883, KP4640, 101488, and 101712. Copyright © 2015 Arivett et al.

  10. Chromosomal Integration of the Klebsiella pneumoniae Carbapenemase Gene, blaKPC, in Klebsiella Species Is Elusive but Not Rare.

    PubMed

    Mathers, Amy J; Stoesser, Nicole; Chai, Weidong; Carroll, Joanne; Barry, Katie; Cherunvanky, Anita; Sebra, Robert; Kasarskis, Andrew; Peto, Tim E; Walker, A Sarah; Sifri, Costi D; Crook, Derrick W; Sheppard, Anna E

    2017-03-01

    Carbapenemase genes in Enterobacteriaceae are mostly described as being plasmid associated. However, the genetic context of carbapenemase genes is not always confirmed in epidemiological surveys, and the frequency of their chromosomal integration therefore is unknown. A previously sequenced collection of blaKPC-positive Enterobacteriaceae from a single U.S. institution (2007 to 2012; n = 281 isolates from 182 patients) was analyzed to identify chromosomal insertions of Tn4401, the transposon most frequently harboring blaKPC Using a combination of short- and long-read sequencing, we confirmed five independent chromosomal integration events from 6/182 (3%) patients, corresponding to 15/281 (5%) isolates. Three patients had isolates identified by perirectal screening, and three had infections which were all successfully treated. When a single copy of blaKPC was in the chromosome, one or both of the phenotypic carbapenemase tests were negative. All chromosomally integrated blaKPC genes were from Klebsiella spp., predominantly K. pneumoniae clonal group 258 (CG258), even though these represented only a small proportion of the isolates. Integration occurred via IS15-ΔI-mediated transposition of a larger, composite region encompassing Tn4401 at one locus of chromosomal integration, seen in the same strain (K. pneumoniae ST340) in two patients. In summary, we identified five independent chromosomal integrations of blaKPC in a large outbreak, demonstrating that this is not a rare event. blaKPC was more frequently integrated into the chromosome of epidemic CG258 K. pneumoniae lineages (ST11, ST258, and ST340) and was more difficult to detect by routine phenotypic methods in this context. The presence of chromosomally integrated blaKPC within successful, globally disseminated K. pneumoniae strains therefore is likely underestimated.

  11. Chromosomal Integration of the Klebsiella pneumoniae Carbapenemase Gene, blaKPC, in Klebsiella Species Is Elusive but Not Rare

    PubMed Central

    Chai, Weidong; Carroll, Joanne; Barry, Katie; Cherunvanky, Anita; Sebra, Robert; Kasarskis, Andrew; Peto, Tim E.; Walker, A. Sarah; Sifri, Costi D.; Crook, Derrick W.; Sheppard, Anna E.

    2016-01-01

    ABSTRACT Carbapenemase genes in Enterobacteriaceae are mostly described as being plasmid associated. However, the genetic context of carbapenemase genes is not always confirmed in epidemiological surveys, and the frequency of their chromosomal integration therefore is unknown. A previously sequenced collection of blaKPC-positive Enterobacteriaceae from a single U.S. institution (2007 to 2012; n = 281 isolates from 182 patients) was analyzed to identify chromosomal insertions of Tn4401, the transposon most frequently harboring blaKPC. Using a combination of short- and long-read sequencing, we confirmed five independent chromosomal integration events from 6/182 (3%) patients, corresponding to 15/281 (5%) isolates. Three patients had isolates identified by perirectal screening, and three had infections which were all successfully treated. When a single copy of blaKPC was in the chromosome, one or both of the phenotypic carbapenemase tests were negative. All chromosomally integrated blaKPC genes were from Klebsiella spp., predominantly K. pneumoniae clonal group 258 (CG258), even though these represented only a small proportion of the isolates. Integration occurred via IS15-ΔI-mediated transposition of a larger, composite region encompassing Tn4401 at one locus of chromosomal integration, seen in the same strain (K. pneumoniae ST340) in two patients. In summary, we identified five independent chromosomal integrations of blaKPC in a large outbreak, demonstrating that this is not a rare event. blaKPC was more frequently integrated into the chromosome of epidemic CG258 K. pneumoniae lineages (ST11, ST258, and ST340) and was more difficult to detect by routine phenotypic methods in this context. The presence of chromosomally integrated blaKPC within successful, globally disseminated K. pneumoniae strains therefore is likely underestimated. PMID:28031204

  12. Clonal dissemination of multilocus sequence type 11 Klebsiella pneumoniae carbapenemase - producing K. pneumoniae in a Chinese teaching hospital.

    PubMed

    Sun, Kangde; Chen, Xu; Li, Chunsheng; Yu, Zhongmin; Zhou, Qi; Yan, Yuzhong

    2015-02-01

    Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae has disseminated rapidly in China. We aimed to analyze the molecular epidemiology of four KPC-producing K. pneumoniae strains isolated from a suspected clonal outbreak during a 3-month period and to track the dissemination of KPC-producing K. pneumonia retrospectively. We created antimicrobial susceptibility profiles using an automated broth microdilution system and broth microdilution methods. We screened carbapenemase and KPC phenotypes using the modified Hodge test and meropenem-boronic acid (BA) disk test, respectively. We identified β-lactamase genes with PCR and sequencing. We investigated clonal relatedness for epidemiological comparison using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). All isolates expressed multidrug resistance and yielded positive results for the modified Hodge and meropenem-BA disk tests. The isolates all carried blaKPC -2 , and coproduced CTX-M-type extended-spectrum β-lactamase. PFGE and MLST showed that the isolates were clonally related. The PFGE patterns of these isolates had ≥90% similarity. We found a single clone, sequence type (ST) 11, and its typical dissemination mode resembled clonal spread. The dissemination of KPC-producing K. pneumoniae is clonally related and there is probable local transmission of a successful ST11 clone.

  13. Pleuritis and suppurative pneumonia associated with a hypermucoviscosity phenotype of Klebsiella pneumoniae in California sea lions (Zalophus californianus).

    PubMed

    Jang, Spencer; Wheeler, Liz; Carey, Roberta B; Jensen, Bette; Crandall, Claudia M; Schrader, Kimmi N; Jessup, David; Colegrove, Kathleen; Gulland, Frances M D

    2010-02-24

    The aim of this study is to document the isolation of a hypermucoviscosity (HMV) phenotype of Klebsiella pneumoniae from 25 cases of suppurative pneumonia and pleuritis and two cases of abscesses in California sea lions (Zalophus californianus) from the central California coast, representing the first report of this zoonotic pathogen from the marine environment and only the second report in non-humans. Animals died 2h to 4 days after first being observed sick on beaches. Clinical signs varied from dyspnoea to coma. Gross post-mortem examination of 25 cases revealed fibrinous pleuritis, copious pus in the pleural cavity and suppurative bronchopneumonia. K. pneumoniae isolates obtained from lung and pleural swabs and the hepatic and subcuticular abscesses were highly mucoid on blood agar culture media and were positive to the "string test". Twenty-one of the 27 isolates were examined by PCR and all were positive for rmpA and K2wyz and negative for K1magA genes. Although pneumonia and pleuritis have previously commonly been observed in marine mammals, their association with pure cultures of a zoonotic bacteria, K. pneumoniae HMV phenotype, has not. This report provides further evidence of the role marine mammals play as sentinels of health risks to humans from coastal waters. Copyright 2009 Elsevier B.V. All rights reserved.

  14. Acute Klebsiella pneumoniae pneumonia alone and with concurrent infection: comparison of clinical and thin-section CT findings.

    PubMed

    Okada, F; Ando, Y; Honda, K; Nakayama, T; Ono, A; Tanoue, S; Maeda, T; Mori, H

    2010-10-01

    The purpose of this study was to identify the clinical and thin-section CT findings in patients with acute Klebsiella pneumoniae pneumonia (KPP) alone and with concurrent infection. We retrospectively identified 160 patients with acute KPP who underwent chest thin-section CT examinations between August 1998 and August 2008 at our institution. The study group comprised 80 patients (54 male, 26 female; age range 18-97 years, mean age 61.5) with acute KPP alone, 55 (43 male, 12 female; age range 46-92 years, mean age 76.0) with KPP combined with methicillin-resistant Staphylococcus aureus (MRSA) and 25 (23 male, 2 female; age range 56-91 years, mean age 72.7) with KPP combined with Pseudomonas aeruginosa (PA). Underlying diseases in patients with each type of pneumonia were assessed. Parenchymal abnormalities were evaluated along with enlarged lymph nodes and pleural effusion. In patients with concurrent pneumonia, underlying conditions such as cardiac diseases, diabetes mellitus and malignancy were significantly more frequent than in patients with KPP alone. The mortality rate in patients with KPP combined with MRSA or PA was significantly higher than in those with KPP alone. In concurrent KPP, CT findings of centrilobular nodules, bronchial wall thickening, cavity, bronchiectasis, nodules and pleural effusion were significantly more frequent with concurrent pneumonia than in those with KPP alone.

  15. Dusuqing granules (DSQ) suppress inflammation in Klebsiella pneumonia rat via NF-κB/MAPK signaling.

    PubMed

    Mei, Xue; Wang, Hao-Xun; Li, Jian-Sheng; Liu, Xiao-Hui; Lu, Xiao-Fan; Li, Ya; Zhang, Wei-Yu; Tian, Yan-Ge

    2017-04-17

    Dusuqing granules (DSQ) have been used in the treatment of bacterial pneumonia clinically, with remarkable benefits. This study was initiated to explore the effects of DSQ on pulmonary inflammation by regulating nuclear factor (NF)-κB/mitogen-activated protein kinase (MAPK) signaling in bacterial pneumonia rats. Rat model was duplicated with Klebsiella pneumonia by a one-time intratracheal injection. Rats were randomized into control, model, DSQ and levofloxacin (LVX) groups. After administrated with appropriate medicines for 7 days, lung tissues were harvested and prepared for pathological analysis, and interleukin (IL)-1, IL-6, monocyte chemotactic protein (MCP)-1and macrophage inflammatory protein (MIP)-2 detections. NF-κB mRNA was measured by real-time qPCR, and the phosphorylation and total proteins of P38MAPK, JNK46/54, ERK42/44 were determined by Western blotting. Marked pathological impairments were observed in model rats, whereas were improved in DSQ group. The cytokines levels, NF-κB mRNA expression and the phosphorylation of P38MAPK, JNK46/54 and ERK42/44 proteins were significantly higher in model group, and were significantly depressed in DSQ group. The protective effects of DSQ on Klebsiella pneumonia might be attributed to its inactivative effects of NF-κB/ MAPK pathway.

  16. Phagocytosis and Killing of Carbapenem-Resistant ST258 Klebsiella pneumoniae by Human Neutrophils.

    PubMed

    Kobayashi, Scott D; Porter, Adeline R; Dorward, David W; Brinkworth, Amanda J; Chen, Liang; Kreiswirth, Barry N; DeLeo, Frank R

    2016-05-15

    Carbapenem-resistant Klebsiella pneumoniae strains classified as multilocus sequence type 258 (ST258) are among the most widespread multidrug-resistant hospital-acquired pathogens. Treatment of infections caused by these organisms is difficult, and mortality is high. The basis for the success of ST258, outside of antibiotic resistance, remains incompletely determined. Here we tested the hypothesis that ST258K. pneumoniae has enhanced capacity to circumvent killing by human neutrophils, the primary cellular defense against bacterial infections. There was limited binding and uptake of ST258 by human neutrophils, and correspondingly, there was limited killing of bacteria. On the other hand, transmission electron microscopy revealed that any ingested organisms were degraded readily within neutrophil phagosomes, thus indicating that survival in the neutrophil assays is due to limited phagocytosis, rather than to microbicide resistance after uptake. Our findings suggest that enhancing neutrophil phagocytosis is a potential therapeutic approach for treatment of infection caused by carbapenem-resistant ST258K. pneumoniae.

  17. First Report of KPC-2 Carbapenemase-Producing Klebsiella pneumoniae in Japan

    PubMed Central

    Takahashi, Rieko; Sawabe, Etsuko; Koyano, Saho; Takahashi, Yutaka; Shima, Mari; Ushizawa, Hiroto; Fujie, Toshihide; Tosaka, Naoki; Kato, Yuko; Moriya, Kyoji; Tohda, Shuji; Tojo, Naoko; Koike, Ryuji; Kubota, Tetsuo

    2014-01-01

    We investigated a novel Japanese isolate of sequence type 11 (ST11), the Klebsiella pneumoniae carbapenemase-2 (KPC-2)-producing K. pneumoniae strain Kp3018, which was previously obtained from a patient treated at a Brazilian hospital. This strain was resistant to various antibiotic classes, including carbapenems, and harbored the gene blaKPC-2, which was present on the transferable plasmid of ca. 190 kb, in addition to the blaCTX-M-15 gene. Furthermore, the ca. 2.3-kb sequences (ISKpn8-blaKPC-2–ISKpn6-like), encompassing blaKPC-2, were found to be similar to those of K. pneumoniae strains from China. PMID:24566171

  18. Tracking a hospital outbreak of carbapenem-resistant Klebsiella pneumoniae with whole-genome sequencing.

    PubMed

    Snitkin, Evan S; Zelazny, Adrian M; Thomas, Pamela J; Stock, Frida; Henderson, David K; Palmore, Tara N; Segre, Julia A

    2012-08-22

    The Gram-negative bacteria Klebsiella pneumoniae is a major cause of nosocomial infections, primarily among immunocompromised patients. The emergence of strains resistant to carbapenems has left few treatment options, making infection containment critical. In 2011, the U.S. National Institutes of Health Clinical Center experienced an outbreak of carbapenem-resistant K. pneumoniae that affected 18 patients, 11 of whom died. Whole-genome sequencing was performed on K. pneumoniae isolates to gain insight into why the outbreak progressed despite early implementation of infection control procedures. Integrated genomic and epidemiological analysis traced the outbreak to three independent transmissions from a single patient who was discharged 3 weeks before the next case became clinically apparent. Additional genomic comparisons provided evidence for unexpected transmission routes, with subsequent mining of epidemiological data pointing to possible explanations for these transmissions. Our analysis demonstrates that integration of genomic and epidemiological data can yield actionable insights and facilitate the control of nosocomial transmission.

  19. Massive pulmonary gangrene: a severe complication of Klebsiella pneumonia.

    PubMed Central

    Knight, L.; Fraser, R. G.; Robson, H. G.

    1975-01-01

    Summary: Massive pulmonary gangrene developed in two patients. Review of the literature reveals 10 other case reports of pulmonary gangrene complicating lobar pneumonia. Among the total of 12 patients whose cases have now been reported, all 4 patients who were treated nonsurgically died and the 8 who underwent surgical resection of the gangrenous lung survived. The present report emphasizes the necessity of early recognition and appropriate surgical treatment for a successful outcome. Images FIG. 1A FIG. 1B FIG. 2 FIG. 3A FIG. 3B FIG. 4 PMID:1089466

  20. Intermingled Klebsiella pneumoniae Populations Between Retail Meats and Human Urinary Tract Infections

    PubMed Central

    Davis, Gregg S.; Waits, Kara; Nordstrom, Lora; Weaver, Brett; Aziz, Maliha; Gauld, Lori; Grande, Heidi; Bigler, Rick; Horwinski, Joseph; Porter, Stephen; Stegger, Marc; Johnson, James R.; Liu, Cindy M.; Price, Lance B.

    2015-01-01

    Background. Klebsiella pneumoniae is a common colonizer of the gastrointestinal tract of humans, companion animals, and livestock. To better understand potential contributions of foodborne K. pneumoniae to human clinical infections, we compared K. pneumoniae isolates from retail meat products and human clinical specimens to assess their similarity based on antibiotic resistance, genetic relatedness, and virulence. Methods. Klebsiella pneumoniae was isolated from retail meats from Flagstaff grocery stores in 2012 and from urine and blood specimens from Flagstaff Medical Center in 2011–2012. Isolates underwent antibiotic susceptibility testing and whole-genome sequencing. Genetic relatedness of the isolates was assessed using multilocus sequence typing and phylogenetic analyses. Extraintestinal virulence of several closely related meat-source and urine isolates was assessed using a murine sepsis model. Results. Meat-source isolates were significantly more likely to be multidrug resistant and resistant to tetracycline and gentamicin than clinical isolates. Four sequence types occurred among both meat-source and clinical isolates. Phylogenetic analyses confirmed close relationships among meat-source and clinical isolates. Isolates from both sources showed similar virulence in the mouse sepsis model. Conclusions. Meat-source K. pneumoniae isolates were more likely than clinical isolates to be antibiotic resistant, which could reflect selective pressures from antibiotic use in food-animal production. The close genetic relatedness of meat-source and clinical isolates, coupled with similarities in virulence, suggest that the barriers to transmission between these 2 sources are low. Taken together, our results suggest that retail meat is a potential vehicle for transmitting virulent, antibiotic-resistant K. pneumoniae from food animals to humans. PMID:26206847

  1. Intermingled Klebsiella pneumoniae Populations Between Retail Meats and Human Urinary Tract Infections.

    PubMed

    Davis, Gregg S; Waits, Kara; Nordstrom, Lora; Weaver, Brett; Aziz, Maliha; Gauld, Lori; Grande, Heidi; Bigler, Rick; Horwinski, Joseph; Porter, Stephen; Stegger, Marc; Johnson, James R; Liu, Cindy M; Price, Lance B

    2015-09-15

    Klebsiella pneumoniae is a common colonizer of the gastrointestinal tract of humans, companion animals, and livestock. To better understand potential contributions of foodborne K. pneumoniae to human clinical infections, we compared K. pneumoniae isolates from retail meat products and human clinical specimens to assess their similarity based on antibiotic resistance, genetic relatedness, and virulence. Klebsiella pneumoniae was isolated from retail meats from Flagstaff grocery stores in 2012 and from urine and blood specimens from Flagstaff Medical Center in 2011-2012. Isolates underwent antibiotic susceptibility testing and whole-genome sequencing. Genetic relatedness of the isolates was assessed using multilocus sequence typing and phylogenetic analyses. Extraintestinal virulence of several closely related meat-source and urine isolates was assessed using a murine sepsis model. Meat-source isolates were significantly more likely to be multidrug resistant and resistant to tetracycline and gentamicin than clinical isolates. Four sequence types occurred among both meat-source and clinical isolates. Phylogenetic analyses confirmed close relationships among meat-source and clinical isolates. Isolates from both sources showed similar virulence in the mouse sepsis model. Meat-source K. pneumoniae isolates were more likely than clinical isolates to be antibiotic resistant, which could reflect selective pressures from antibiotic use in food-animal production. The close genetic relatedness of meat-source and clinical isolates, coupled with similarities in virulence, suggest that the barriers to transmission between these 2 sources are low. Taken together, our results suggest that retail meat is a potential vehicle for transmitting virulent, antibiotic-resistant K. pneumoniae from food animals to humans. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America.

  2. Surfactant Protein D Enhances Phagocytosis and Killing of Unencapsulated Phase Variants of Klebsiella pneumoniae

    PubMed Central

    Ofek, Itzhak; Mesika, Adi; Kalina, Moshe; Keisari, Yona; Podschun, Ranier; Sahly, Hany; Chang, Donald; McGregor, David; Crouch, Erika

    2001-01-01

    Pulmonary surfactant protein D (SP-D) is a collagenous C-type lectin (collectin) that is secreted into the alveoli and distal airways of the lung. We have studied the interactions of SP-D and alveolar macrophages with Klebsiella pneumoniae, a common cause of nosocomial pneumonia. SP-D does not agglutinate encapsulated K. pneumoniae but selectively agglutinates spontaneous, unencapsulated phase variants, such as Klebsiella strain K50-3OF, through interactions with their lipopolysaccharides (LPS). These effects are calcium dependent and inhibited with maltose but not lactose, consistent with involvement of the SP-D carbohydrate recognition domain. Precoating of K50-3OF with SP-D enhances the phagocytosis and killing of these organisms by rat alveolar macrophages in cell culture and stimulates the production of nitric oxide by the NR-8383 rat alveolar macrophage cell line. SP-D similarly enhances the NO response to K50-3OF LPS adsorbed to Latex beads under conditions where soluble LPS or SP-D, or soluble complexes of SP-D and LPS, do not stimulate NO production. Our studies demonstrate that interactions of SP-D with exposed arrays of Klebsiella LPS on a particulate surface can enhance the host defense activities of alveolar macrophages and suggest that activation of macrophages by SP-D requires binding to microorganisms or other particulate ligands. Because unencapsulated phase variants are likely to be responsible for the initial stages of tissue invasion and infection, we speculate that SP-D-mediated agglutination and/or opsonization of K. pneumoniae is an important defense mechanism for this respiratory pathogen in otherwise healthy individuals. PMID:11119485

  3. Adherence properties of an mrkD-negative mutant of Klebsiella pneumoniae.

    PubMed Central

    Hornick, D B; Thommandru, J; Smits, W; Clegg, S

    1995-01-01

    The role of the mrkD gene in attachment by a type 3 fimbriate Klebsiella pneumoniae strain was further characterized. A clinical isolate, K. pneumoniae IA565, was found to contain two copies of the gene encoding the fimbrial subunit, mrkA, and one copy of the gene encoding the adhesin subunit, mrkD. One copy of mrkA was located on the bacterial chromosome, and the other copy was associated with mrkD and located on a plasmid. The plasmid-borne mrk gene cluster was lost when K. pneumoniae IA565 was subcultured serially in broth at 44 degrees C. The resulting mrkD-negative strain, designated K. pneumoniae IApc35, did not exhibit the following adherence characteristics associated with K. pneumoniae possessing MrkD-positive fimbriae: agglutination of tannic acid-treated human erythrocytes and attachment to trypsinized human buccal cells. However, K. pneumoniae IApc35 produced type 3 fimbriae that were composed of the characteristic 21.5-kDa major fimbrial subunit, were reactive with specific serum, and were visualized specifically by immunoelectron microscopy. K. pneumoniae IApc35 retained a copy of the mrkA gene on its chromosome. This mrkA-containing gene cluster could be complemented by a recombinant plasmid carrying only the mrkD gene, resulting in restoration of the K. pneumoniae IA565-like adhesive phenotype and demonstration of type 3 filament-associated MrkD subunits by using colloidal gold labeling and immunoelectron microscopy. These data indicate that K. pneumoniae may contain multiple copies of the mrk genes which may be present simultaneously on both plasmid and chromosomal DNAs and which may encode fimbriae with different binding specificities. PMID:7729917

  4. Retrospective investigation of the clinical effects of tazobactam/piperacillin and sulbactam/ampicillin on aspiration pneumonia caused by Klebsiella pneumoniae.

    PubMed

    Tsukada, Hiroki; Sakai, Kunihiko; Cho, Hiromi; Kimura, Yuka; Tetsuka, Takafumi; Nakajima, Haruhiko; Ito, Kazuhiko

    2012-10-01

    Klebsiella pneumoniae is an important causative bacterium of aspiration pneumonia in many elderly patients. We retrospectively investigated the clinical effects of the early treatment of aspiration pneumonia and background factors in 24 patients from whom Klebsiella pneumoniae was isolated. Sulbactam/ampicillin (SBT/ABPC) was selected for early treatment in 12 of the 24 patients diagnosed with aspiration pneumonia, and tazobactam/piperacillin (TAZ/PIPC) was selected for the other patients. The effective rates and success rates of early treatment were significantly higher in the TAZ/PIPC group than in the SBT/ABPC group (p = 0.003 and 0.027, respectively). Although no significant difference was noted because of the limited number of cases, the survival rates after 30 days were 91.7 and 58.3 % in the TAZ/PIPC and SBT/ABPC groups, respectively. Several bacteria isolated with Klebsiella pneumoniae were resistant bacteria, such as methicillin-resistant Staphylococcus aureus or Pseudomonas aeruginosa, and no anaerobe or extended-spectrum β-lactamase-producing Klebsiella pneumoniae was isolated. Thirteen and 11 of the 24 cases were classified as healthcare-associated pneumonia (HCAP) and hospital-acquired pneumonia (HAP), respectively, with no case classified as community-acquired pneumonia (CAP). As population aging progresses, the frequency of aspiration pneumonia classified as HCAP will increase. To cover anaerobes, it is necessary to select antibacterial drugs, such as TAZ/PIPC, for early treatment in consideration of resistant gram-negative bacteria to improve the outcome, and not drugs with weak activity against these bacteria.

  5. A high-resolution genomic analysis of multidrug-resistant hospital outbreaks of Klebsiella pneumoniae

    PubMed Central

    Chung The, Hao; Karkey, Abhilasha; Pham Thanh, Duy; Boinett, Christine J; Cain, Amy K; Ellington, Matthew; Baker, Kate S; Dongol, Sabina; Thompson, Corinne; Harris, Simon R; Jombart, Thibaut; Le Thi Phuong, Tu; Tran Do Hoang, Nhu; Ha Thanh, Tuyen; Shretha, Shrijana; Joshi, Suchita; Basnyat, Buddha; Thwaites, Guy; Thomson, Nicholas R; Rabaa, Maia A; Baker, Stephen

    2015-01-01

    Multidrug-resistant (MDR) Klebsiella pneumoniae has become a leading cause of nosocomial infections worldwide. Despite its prominence, little is known about the genetic diversity of K. pneumoniae in resource-poor hospital settings. Through whole-genome sequencing (WGS), we reconstructed an outbreak of MDR K. pneumoniae occurring on high-dependency wards in a hospital in Kathmandu during 2012 with a case-fatality rate of 75%. The WGS analysis permitted the identification of two MDR K. pneumoniae lineages causing distinct outbreaks within the complex endemic K. pneumoniae. Using phylogenetic reconstruction and lineage-specific PCR, our data predicted a scenario in which K. pneumoniae, circulating for 6 months before the outbreak, underwent a series of ward-specific clonal expansions after the acquisition of genes facilitating virulence and MDR. We suggest that the early detection of a specific NDM-1 containing lineage in 2011 would have alerted the high-dependency ward staff to intervene. We argue that some form of real-time genetic characterisation, alongside clade-specific PCR during an outbreak, should be factored into future healthcare infection control practices in both high- and low-income settings. PMID:25712531

  6. Identification and characterization of antigens as vaccine candidates against Klebsiella pneumoniae

    PubMed Central

    Lundberg, Urban; Senn, Beatrice M.; Schüler, Wolfgang; Meinke, Andreas; Hanner, Markus

    2013-01-01

    Nosocomial infections, also called “hospital acquired infections,” occur worldwide and affect both developed and resource-poor countries, thus having a major impact on their health care systems. Klebsiella pneumoniae, which is an opportunistic Gram-negative pathogen, is responsible for causing pneumonia, urinary tract infections and septicemia in immune compromised hosts such as neonates. Unfortunately, there is no vaccine or mAb available for prophylactic or therapeutic use against K. pneumoniae infections. For this reason, we sought for a protein-based subunit vaccine capable of combating K. pneumoniae infections, by applying our ANTIGENome technology for the identification of potential vaccine candidates, focusing on conserved protein antigens present in strains with different serotypes. We identified numerous novel immunogenic proteins using genomic surface display libraries and human serum antibodies from donors exposed to or infected by K. pneumoniae. Vaccine candidate antigens were finally selected based on animal protection in a murine lethal-sepsis model. The protective and highly conserved antigens identified in this study are promising candidates for the development of a protein-based vaccine to prevent infection by K. pneumoniae. PMID:23250007

  7. A high-resolution genomic analysis of multidrug-resistant hospital outbreaks of Klebsiella pneumoniae.

    PubMed

    Chung The, Hao; Karkey, Abhilasha; Pham Thanh, Duy; Boinett, Christine J; Cain, Amy K; Ellington, Matthew; Baker, Kate S; Dongol, Sabina; Thompson, Corinne; Harris, Simon R; Jombart, Thibaut; Le Thi Phuong, Tu; Tran Do Hoang, Nhu; Ha Thanh, Tuyen; Shretha, Shrijana; Joshi, Suchita; Basnyat, Buddha; Thwaites, Guy; Thomson, Nicholas R; Rabaa, Maia A; Baker, Stephen

    2015-03-01

    Multidrug-resistant (MDR) Klebsiella pneumoniae has become a leading cause of nosocomial infections worldwide. Despite its prominence, little is known about the genetic diversity of K. pneumoniae in resource-poor hospital settings. Through whole-genome sequencing (WGS), we reconstructed an outbreak of MDR K. pneumoniae occurring on high-dependency wards in a hospital in Kathmandu during 2012 with a case-fatality rate of 75%. The WGS analysis permitted the identification of two MDR K. pneumoniae lineages causing distinct outbreaks within the complex endemic K. pneumoniae. Using phylogenetic reconstruction and lineage-specific PCR, our data predicted a scenario in which K. pneumoniae, circulating for 6 months before the outbreak, underwent a series of ward-specific clonal expansions after the acquisition of genes facilitating virulence and MDR. We suggest that the early detection of a specific NDM-1 containing lineage in 2011 would have alerted the high-dependency ward staff to intervene. We argue that some form of real-time genetic characterisation, alongside clade-specific PCR during an outbreak, should be factored into future healthcare infection control practices in both high- and low-income settings.

  8. An unusual culprit: Klebsiella pneumoniae causing septicaemia outbreaks in neonatal pigs?

    PubMed

    Bowring, Bethany G; Fahy, V Anthony; Morris, Andrew; Collins, Alison M

    2017-05-01

    This study investigated the cause of recent outbreaks of septicaemia in neonatal pigs in Australia (Victoria and Queensland). Septicaemia in neonatal pigs is commonly caused by enterotoxigenic E. coli, extraintestinal pathogenic E. coli and beta-haemolytic streptococci. Infrequent causes of septicaemia are Actinobacillus suis and Citrobacter freundii. Therefore, it was quite unexpected when Klebsiella pneumoniae was isolated in predominant growth from multiple organs of pigs dying of septicaemia from six days of age. Two cases in Victoria were associated with multilocus sequence type 25 (ST25), whereas the cases on a single farm in Queensland were associated with two different sequence types (ST278 and ST1978). Similar outbreaks of septicaemia have also occurred in England, but all were associated with K. pneumoniae ST25. The K. pneumoniae sequence types did not cluster phylogenetically into groups of isolates from the same geographical location or into groups which caused either septicaemia or pneumonia. Antibiotic resistance also varied between isolates and showed neomycin resistance in Queensland. These results suggest that several sequence types of K. pneumoniae are involved in causing outbreaks of septicaemia in neonatal pigs, in addition to its previously recognised role in pneumonia and mastitis. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Prevalence of Acquired Carbapenemase Genes in Klebsiella Pneumoniae by Multiplex PCR in Isfahan

    PubMed Central

    Khorvash, Farzin; Yazdani, Mohammed Reza; Soudi, Ali Asghar; Shabani, Shiva; Tavahen, Nirvana

    2017-01-01

    Background: Multi-drug resistant Klebsiella pneumoniae has been considered as a serious global threat. This study was done to investigate carbapenemase producing genomes among K. pneumoniae isolates in Isfahan, Central Iran. Materials and Methods: In a cross-sectional study from 2011 to 2012, 29 carbapenem resistant (according to disc diffusion method) carbapenemase producing (according to modified Hodge test) K. pneumoniae strains were collected from Intensive Care Unit (ICUs) of Al-Zahra referral Hospital. In the strains with the lack of sensitivity to one or several carbapenems, beta-lactams, or beta-lactamases, there has been performed modified Hodge test to investigate carbapenmase and then only strains producing carbapenmases were selected for molecular methods. Results: In this study, there have been 29 cases of K. pneumoniae isolated from hospitalized patients in the (ICU). Three cases (10.3%) contained blaVIM, 1 case (3.4%) contained blaIMP, and 1 case (3.4%) contained blaOXA. The genes blaNDM and blaKPC were not detected. Then, 16 cases (55.2%) from positive cases of K. pneumoniae were related to the chip, 4 cases (13.8%) to catheter, 6 cases (20.7%) to urine, and 3 cases (10.3%) to wound. Conclusion: It is necessary to monitor the epidemiologic changes of these carbapenemase genes in K. pneumoniae in our Hospital. More attention should be paid to nosocomial infection control measures. Other carbapenemase producing genes should be investigated. PMID:28503496

  10. Prevalence of Acquired Carbapenemase Genes in Klebsiella Pneumoniae by Multiplex PCR in Isfahan.

    PubMed

    Khorvash, Farzin; Yazdani, Mohammed Reza; Soudi, Ali Asghar; Shabani, Shiva; Tavahen, Nirvana

    2017-01-01

    Multi-drug resistant Klebsiella pneumoniae has been considered as a serious global threat. This study was done to investigate carbapenemase producing genomes among K. pneumoniae isolates in Isfahan, Central Iran. In a cross-sectional study from 2011 to 2012, 29 carbapenem resistant (according to disc diffusion method) carbapenemase producing (according to modified Hodge test) K. pneumoniae strains were collected from Intensive Care Unit (ICUs) of Al-Zahra referral Hospital. In the strains with the lack of sensitivity to one or several carbapenems, beta-lactams, or beta-lactamases, there has been performed modified Hodge test to investigate carbapenmase and then only strains producing carbapenmases were selected for molecular methods. In this study, there have been 29 cases of K. pneumoniae isolated from hospitalized patients in the (ICU). Three cases (10.3%) contained blaVIM, 1 case (3.4%) contained blaIMP, and 1 case (3.4%) contained blaOXA. The genes blaNDM and blaKPC were not detected. Then, 16 cases (55.2%) from positive cases of K. pneumoniae were related to the chip, 4 cases (13.8%) to catheter, 6 cases (20.7%) to urine, and 3 cases (10.3%) to wound. It is necessary to monitor the epidemiologic changes of these carbapenemase genes in K. pneumoniae in our Hospital. More attention should be paid to nosocomial infection control measures. Other carbapenemase producing genes should be investigated.

  11. Genotyping of Klebsiella Pneumonia Strains Isolated from Eldly Inpatients by Multiple-locus Variable-number Tandem-repeat Analysis.

    PubMed

    Zhang, Yuan-Yuan; Xu, Ya-Ping; DU, Peng-Cheng; Qiang, Yu-Jun; Zhang, Wen; Chen, Chen; Yu, Ji-Hong; Guo, Jun

    2016-08-01

    Objective To investigate the genotype of klebsiella pneumonia strains isolated from eldly inpatients by multiple-locus variable-number tandem-repeat analysis. Methods Totally 184 klebsiella pneumonia strains,isolated from eldly inpatients,were collected,and their genome DNA were extracted. The polymorphism of 7 variable-number tandem-repeat locus in the DNA samples was analyzed by multiple primers polymerase chain reaction and capillary electrophoresis. The clustering analysis of genotyping was carried out with the BioNumerics 5.1 software. Results A total of 139 genotypes were identified in 184 klebsiella pneumonia clinical strains,showing obvious genetic polymorphisms. With clustering analysis of genotypes,all the strains were categorized into three gene clusters (genogroups 1,2,and 3). The genogroup 1 was the biggest cluster,containing 93.06% of the isolated strains. Conclusion There was a predominant cluster in the klebsiella pneumonia strains isolated from eldly inpatients in our center,and the major source of klebsiella pneumonia infection remained the nosocomial infection.

  12. Bacteriophage-loaded nanostructured lipid carrier: improved pharmacokinetics mediates effective resolution of Klebsiella pneumoniae-induced lobar pneumonia.

    PubMed

    Singla, Saloni; Harjai, Kusum; Katare, Om Prakash; Chhibber, Sanjay

    2015-07-15

    This study examined the therapeutic and prophylactic potential of bacteriophages in a mouse model of Klebsiella pneumoniae lobar pneumonia. Phages were administered intraperitoneally. Liposome-entrapped phages (LP) were effective in treating infection, even when therapy was delayed by 3 days after the induction of pneumonia. In contrast, nonliposomal phages provided protection when administered 24 hours after infection. Administration of nonliposomal phages 6 hours prior to intranasal bacterial challenge resulted in complete protection, compared with LP, which was effective even when administered 48 hours prior to infection. Increased reduction and a greater increment in the levels of proinflammatory and antiinflammatory cytokines, respectively, in homogenates of lung from LP-treated mice were suggestive of increased efficacy of LP in the treatment of pneumonia. This is the first study to assess liposomes as a delivery vehicle for phage, and the results confirm the superiority of LP for both therapeutic and prophylactic applications. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  13. A comparative study of induction of pneumonia in mice with planktonic and biofilm cells of Klebsiella pneumoniae.

    PubMed

    Sharma, Sonica; Mohan, Harsh; Sharma, Saroj; Chhibber, Sanjay

    2011-05-01

    In the present study, the course of acute pneumonia in normal BALB/c mice infected by intranasal inoculation of planktonic and preformed biofilm cells (3 days old) of Klebsiella pneumoniae B5055 was studied and compared. With both cell forms the peak of infection was observed on the third post infection day, as assessed on the basis of lung bacterial load and corresponding pathology. There was an intense neutrophil infiltration in bronchoalveolar lavage fluid. Tissue damage was assessed on the basis of increased amounts of nitrite, malondialdehyde and lactate dehydrogenase in lung homogenates. The phagocytic potential of alveolar macrophages was lower in biofilm cell-induced infection than in that induced by planktonic cells. Biofilm cell induced infection generated significantly greater production of tumor necrosis factor-α and interleukin-1β on the third and fifth days of infection, respectively. Production of interleukin-10 was, however, variable. There was no significant difference in the ability of planktonic and biofilm cell forms of K. pneumoniae to induce acute pneumonia in mice in terms of bacterial counts and histopathological changes. However, biofilm cell-induced infection showed delayed clearance as compared to infection induced with the planktonic form. © 2011 The Societies and Blackwell Publishing Asia Pty Ltd.

  14. Rapid discrimination of Klebsiella pneumoniae carbapenemase 2-producing and non-producing Klebsiella pneumoniae strains using near-infrared spectroscopy (NIRS) and multivariate analysis.

    PubMed

    Marques, Aline S; Moraes, Edgar P; Júnior, Miguel A A; Moura, Andrew D; Neto, Valter F A; Neto, Renato M; Lima, Kássio M G

    2015-03-01

    Klebsiella pneumoniae Carbapenemase (KPC-2)-producing and non-producing Klebsiella pneumoniae (KP) have rapidly disseminated worldwide, challenging the diagnostics of Gram-negative infections. We evaluate the potential of a novel non-destructive and rapid method based on Near-Infrared Spectroscopic (NIRS) and multivariate analysis for distinguishing KPC-2-producing and non-producing KP. Thirty-nine NIRS spectra (24 KPC-2-producing KP, 15 KPC-2 non-producing KP) were acquired; different pre-processing methods such as baseline correction, derivative and Savitzky-Golay smoothing were performed. A spectral region fingerprint was achieved after using genetic algorithm-linear discriminant analysis (GA-LDA) and successive projection algorithm (SPA-LDA) algorithms for variable selection. The variables selected were then used for discriminating the microorganisms.Accuracy test results including sensitivity and specificity were determined. Sensitivity in KPC-2 producing and non-producing KP categories was 66.7% and 75%, respectively, using a SPA-LDA model with 66 wavenumbers. The resulting GA-LDA model successfully classified both microorganisms with respect to their "fingerprints" using only 39 wavelengths. Sensitivity in KPC-2 producing category was moderate(≈66.7%) using a GA-LDA model. However, sensitivity in KPC-2 non-producing category using GA-LDA accurately predicted the correct class (with 100% accuracy). As100% accuracy was achieved, this novel approach identifies potential biochemical markers that may have a relation with microbial functional roles and means of rapid identification of KPC-2 producing and non-producing KP strains.

  15. Outcomes of transplantation using organs from a donor infected with Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae.

    PubMed

    Ariza-Heredia, E J; Patel, R; Blumberg, E A; Walker, R C; Lewis, R; Evans, J; Sankar, A; Willliams, M D; Rogers, J; Milano, C; Razonable, R R

    2012-06-01

    Transmission of pathogens from donor to recipient is a potential complication of organ transplantation. Herein, we describe the clinical course and outcomes of 4 transplant recipients who received tissues from a donor with multi-organ infection with Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae. Recipient 1 underwent simultaneous liver and kidney transplantation for alpha-1 antitrypsin deficiency and alcohol-related cirrhosis, and acute tubular necrosis, respectively. Soon after transplantation, he developed an infected hematoma and peritonitis due to KPC-producing K. pneumoniae despite receiving tigecycline prophylaxis. He was treated with a prolonged course of tigecycline, amikacin, and meropenem, in conjunction with surgical evacuation and percutaneous drainage of the infected fluid collections. Recipient 2 underwent living-donor liver transplantation for cholangiocarcinoma and primary sclerosing cholangitis using vein graft from the donor infected with KPC-producing K. pneumoniae. Culture of the preservation fluid containing the vein graft was positive for KPC-producing K. pneumoniae. The patient received preemptive amikacin and tigecycline, and he did not develop any infection (as evidenced by negative surveillance blood cultures). The isolates from the donor and Recipients 1 and 2 were indistinguishable by pulsed-field gel electrophoresis. Recipients 3 and 4 underwent kidney and heart transplantation, respectively; both patients received perioperative tigecycline prophylaxis and did not develop infections due to KPC-producing K. pneumoniae. All transplant recipients had good short-term outcomes. These cases highlight the importance of inter-institutional communication and collaboration to ensure the successful management of recipients of organs from donors infected with multidrug-resistant organisms.

  16. Topical treatment of Klebsiella pneumoniae B5055 induced burn wound infection in mice using natural products.

    PubMed

    Kumari, Seema; Harjai, Kusum; Chhibber, Sanjay

    2010-06-30

    Burn wound infection remains the principal cause of death in burn patients. Efficacy of honey and aloe vera gel was evaluated in the treatment of burn wound infection induced with Klebsiella pneumoniae B5055 and their efficacy was compared with an isolated and well-characterized Klebsiella specific phage Kpn5. A full thickness burn wound was induced in mice and infected with K. pneumoniae B5055 via topical route. The efficacy of natural antimicrobial agents (honey and aloe vera gel) topically applied daily was compared with the efficacy of phage Kpn5 suspended in hydrogel applied topically a single time on the burn wound. Efficacy of these antimicrobial agents was assessed on the basis of the percentage of infected mice that survived following treatment. In comparison to untreated control mice, those treated with a single dose of phage Kpn5 at MOI of 200 showed significant reduction in mortality (P < 0.001). Daily application of honey and aloe vera gel provided significant protection (P < 0.001), but in combination with phage, no additional advantage was observed (P > 0.05) compared to the use of honey and aloe vera gel alone. The results of this study strongly suggest that phage Kpn5 has therapeutic value in treating burn wound infection in mice as a single topical application of this phage was able to rescue mice from infection caused by K. pneumoniae B5055 in comparison to multiple applications of honey and aloe vera gel.

  17. 2,3-butanediol production from starch by engineered Klebsiella pneumoniae G31-A.

    PubMed

    Tsvetanova, Flora; Petrova, Penka; Petrov, Kaloyan

    2014-03-01

    2,3-Butanediol (2,3-BD) is an organic compound, which is widely used as a fuel and fuel additive and applied in chemical, food, and pharmaceutical industries. Contemporary strategies for its economic synthesis include the development of microbial technologies that use starch as cheap and renewable feedstock. The present work encompasses the metabolic engineering of the excellent 2,3-BD producer Klebsiella pneumoniae G31. In order to perform direct starch conversion into 2,3-BD, the amyL gene encoding quite active, liquefying α-amylase in Bacillus licheniformis was cloned under lac promoter control in the recombinant K. pneumoniae G31-A. The enhanced extracellular over-expression of amyL led to the highest extracellular amylase activity (68 U/ml) ever detected in Klebsiella. The recombinant strain was capable of simultaneous saccharification and fermentation (SSF) of potato starch to 2,3-BD. In SSF batch process by the use of 200 g/l starch, the amount of total diols produced was 60.9 g/l (53.8 g/l 2,3-BD and 7.1 g/l acetoin), corresponding to 0.31 g/g conversion rate. The presented results are the first to show successful starch conversion to 2,3-BD by K. pneumoniae in a one-step process.

  18. [Characterization of SHV-5 beta-lactamase-producing Klebsiella pneumoniae in an intensive care unit].

    PubMed

    Andrade, Verónica; Silva, Jesús

    2004-01-01

    To perform the molecular characterization of Klebsiella pneumoniae isolates from pediatric patients and health care workers at the intensive care unit of a tertiary care hospital in Mexico City. Fifteen Klebsiella pneumoniae isolates collected during an outbreak in June 1996 were analyzed; eight were from patients and seven from health care workers of Mexico's Children's Hospital. Characterization of isolates was carried out by pulsed field gel electrophoresis (PFGE), random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) and serotyping, beta-Lactamase isoelectric focusing (IEF), and nucleotide sequencing of PCR products. Serotype 61 was predominant and correlated with genomic fingerprints of RAPD and PFGE in 11 of 15 isolates. One SHV-5-producer predominant clone with a high case-fatality rate was identified. Molecular biology techniques are useful tools to characterize the K. pneumoniae clone isolated from patients and health care workers, suggesting potential cross-transmission. These data call for strengthening control programs to prevent dissemination of nosocomial infections in the studied hospital.

  19. Comparative effects of carbapenems on bacterial load and host immune response in a Klebsiella pneumoniae murine pneumonia model.

    PubMed

    Hilliard, Jamese J; Melton, John L; Hall, LeRoy; Abbanat, Darren; Fernandez, Jeffrey; Ward, Christine K; Bunting, Rachel A; Barron, A; Lynch, A Simon; Flamm, Robert K

    2011-02-01

    Doripenem is a carbapenem with potent broad-spectrum activity against Gram-negative pathogens, including antibiotic-resistant Enterobacteriaceae. As the incidence of extended-spectrum β-lactamase (ESBL)-producing Gram-negative bacilli is increasing, it was of interest to examine the in vivo comparative efficacy of doripenem, imipenem, and meropenem against a Klebsiella pneumoniae isolate expressing the TEM-26 ESBL enzyme. In a murine lethal lower respiratory infection model, doripenem reduced the Klebsiella lung burden by 2 log(10) CFU/g lung tissue over the first 48 h of the infection. Treatment of mice with meropenem or imipenem yielded reductions of approximately 1.5 log(10) CFU/g during this time period. Seven days postinfection, Klebsiella titers in the lungs of treated mice decreased an additional 2 log(10) CFU/g relative to those in the lungs of untreated control animals. Lipopolysaccharide (LPS) endotoxin release assays indicated that 6 h postinfection, meropenem- and imipenem-treated animals had 10-fold more endotoxin in lung homogenates and sera than doripenem-treated mice. Following doripenem treatment, the maximum endotoxin release postinfection (6 h) was 53,000 endotoxin units (EU)/ml, which was 2.7- and 6-fold lower than imipenem or meropenem-treated animals, respectively. While the levels of several proinflammatory cytokines increased in both the lungs and sera following intranasal K. pneumoniae inoculation, doripenem treatment, but not meropenem or imipenem treatment, resulted in significantly increased interleukin 6 levels in lung homogenates relative to those in lung homogenates of untreated controls, which may contribute to enhanced neutrophil killing of bacteria in the lung. Histological examination of tissue sections indicated less overall inflammation and tissue damage in doripenem-treated mice, consistent with improved antibacterial efficacy, reduced LPS endotoxin release, and the observed cytokine induction profile.

  20. Epithelial Myeloid-differentiation Factor 88 is Dispensable During Klebsiella Pneumonia.

    PubMed

    Anas, Adam A; Claushuis, Theodora A M; Mohan, Rajiv A; Christoffels, Vincent M; Aidinis, Vassilis; Florquin, Sandrine; Van't Veer, Cornelis; Hou, Baidong; de Vos, Alex F; van der Poll, Tom

    2017-02-10

    Klebsiella (K.) pneumoniae is a common cause of pneumonia. Previous studies have documented an important role for Toll-like receptors (TLRs) expressed by myeloid cells in the recognition of K. pneumoniae and the initiation of a protective immune response. Lung epithelial cells also express TLRs and can participate in innate immune defense. The aim of this study was to examine the role of the common TLR adaptor protein myeloid differentiation factor (MyD)88 in lung epithelium during host defense against K. pneumoniae induced pneumonia. For this we first crossed mice expressing cre recombinase under the control of the surfactant protein C (SftpCcre) or the club cell 10Kd (CC10cre) promoter with reporter mice to show that SftpCcre mice mainly express cre in type II alveolar cells, while CC10cre mice express cre almost exclusively in bronchiolar epithelial cells. We then generated mice with cell targeted deletion of MyD88 in type II alveolar (SftpCcre-MyD88-lox) and bronchiolar epithelial (CC10cre-MyD88-lox) cells, and infected them with K. pneumoniae via the airways. Bacterial growth and dissemination were not affected by the loss of MyD88 in SftpCcre-MyD88-lox or CC10cre-MyD88-lox mice compared to control littermates. Furthermore, inflammatory responses induced by K. pneumoniae in the lung were not dependent on MyD88 expression in type II alveolar or bronchiolar epithelial cells. These results indicate that MyD88 expression in two distinct lung epithelial cell types does not contribute to host defense during pneumonia caused by a common human gram-negative pathogen.

  1. Global Dissemination of Carbapenemase-Producing Klebsiella pneumoniae: Epidemiology, Genetic Context, Treatment Options, and Detection Methods

    PubMed Central

    Lee, Chang-Ro; Lee, Jung Hun; Park, Kwang Seung; Kim, Young Bae; Jeong, Byeong Chul; Lee, Sang Hee

    2016-01-01

    The emergence of carbapenem-resistant Gram-negative pathogens poses a serious threat to public health worldwide. In particular, the increasing prevalence of carbapenem-resistant Klebsiella pneumoniae is a major source of concern. K. pneumoniae carbapenemases (KPCs) and carbapenemases of the oxacillinase-48 (OXA-48) type have been reported worldwide. New Delhi metallo-β-lactamase (NDM) carbapenemases were originally identified in Sweden in 2008 and have spread worldwide rapidly. In this review, we summarize the epidemiology of K. pneumoniae producing three carbapenemases (KPCs, NDMs, and OXA-48-like). Although the prevalence of each resistant strain varies geographically, K. pneumoniae producing KPCs, NDMs, and OXA-48-like carbapenemases have become rapidly disseminated. In addition, we used recently published molecular and genetic studies to analyze the mechanisms by which these three carbapenemases, and major K. pneumoniae clones, such as ST258 and ST11, have become globally prevalent. Because carbapenemase-producing K. pneumoniae are often resistant to most β-lactam antibiotics and many other non-β-lactam molecules, the therapeutic options available to treat infection with these strains are limited to colistin, polymyxin B, fosfomycin, tigecycline, and selected aminoglycosides. Although, combination therapy has been recommended for the treatment of severe carbapenemase-producing K. pneumoniae infections, the clinical evidence for this strategy is currently limited, and more accurate randomized controlled trials will be required to establish the most effective treatment regimen. Moreover, because rapid and accurate identification of the carbapenemase type found in K. pneumoniae may be difficult to achieve through phenotypic antibiotic susceptibility tests, novel molecular detection techniques are currently being developed. PMID:27379038

  2. Klebsiella pneumoniae outer membrane protein A is required to prevent the activation of airway epithelial cells.

    PubMed

    March, Catalina; Moranta, David; Regueiro, Verónica; Llobet, Enrique; Tomás, Anna; Garmendia, Junkal; Bengoechea, José A

    2011-03-25

    Outer membrane protein A (OmpA) is a class of proteins highly conserved among the Enterobacteriaceae family and throughout evolution. Klebsiella pneumoniae is a capsulated gram-negative pathogen. It is an important cause of community-acquired and nosocomial pneumonia. Evidence indicates that K. pneumoniae infections are characterized by a lack of an early inflammatory response. Data from our laboratory indicate that K. pneumoniae CPS helps to suppress the host inflammatory response. However, it is unknown whether K. pneumoniae employs additional factors to modulate host inflammatory responses. Here, we report that K. pneumoniae OmpA is important for immune evasion in vitro and in vivo. Infection of A549 and normal human bronchial cells with 52OmpA2, an ompA mutant, increased the levels of IL-8. 52145-Δwca(K2)ompA, which does not express CPS and ompA, induced the highest levels of IL-8. Both mutants could be complemented. In vivo, 52OmpA2 induced higher levels of tnfα, kc, and il6 than the wild type. ompA mutants activated NF-κB, and the phosphorylation of p38, p44/42, and JNK MAPKs and IL-8 induction was via NF-κB-dependent and p38- and p44/42-dependent pathways. 52OmpA2 engaged TLR2 and -4 to activate NF-κB, whereas 52145-Δwca(K2)ompA activated not only TLR2 and TLR4 but also NOD1. Finally, we demonstrate that the ompA mutant is attenuated in the pneumonia mouse model. The results of this study indicate that K. pneumoniae OmpA contributes to attenuate airway cell responses. This may facilitate pathogen survival in the hostile environment of the lung.

  3. Risk factors for KPC-producing Klebsiella pneumoniae: watch out for surgery.

    PubMed

    da Silva, Kesia Esther; Maciel, Wirlaine Glauce; Sacchi, Flávia Patussi Correia; Carvalhaes, Cecilia Godoy; Rodrigues-Costa, Fernanda; da Silva, Ana Carolina Ramos; Croda, Mariana Garcia; Negrão, Fábio Juliano; Croda, Julio; Gales, Ana Cristina; Simionatto, Simone

    2016-06-01

    This study describes the molecular characteristics and risk factors associated with carbapenem-resistant Klebsiella pneumoniae strains. Risk factors associated with KPC-producing K. pneumoniae strains were investigated in this case-control study from May 2011 to May 2013. Bacterial identification was performed by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS). Antimicrobial susceptibility was determined by broth microdilution. Carbapenemase production was assessed by both modified Hodge test (MHT) and ertapenem hydrolysis using MALDI-TOF MS. The presence of β-lactamase-encoding genes was evaluated by PCR and DNA sequencing. Alterations in genes encoding K. pneumoniae outer membrane proteins were analysed by PCR and DNA sequencing as well as SDS-PAGE. Genetic relatedness among strains was determined by pulsed-field gel electrophoresis. This study included 94 patients. Longer hospitalisation, mechanical ventilation, catheters, and previous surgery were associated with KPC-producing K. pneumoniae. Sixty-eight strains showed resistance to carbapenems. Carbapenemase production was detected by MHT in 67 K. pneumoniae strains and by MALDI-TOF MS in 57. The presence of the blaKPC-2 gene was identified in 57 strains. The blaKPC-2 gene was not found in 11 carbapenem-resistant K. pneumoniae; instead, the blaCTX-M-1-like, blaCTX-M-2-like, blaCTX-M-8 like, blaCTX-M-14-like and blaSHV- like genes associated with OmpK35 and OmpK36 alterations were observed. Thirty-three KPC-producing K. pneumoniae strains were clonally related, and patients infected with these strains had a higher mortality rate (78.78 %). Our results show that KPC-producing K. pneumoniae was associated with several healthcare-related risk factors, including recent surgery.

  4. Klebsiella pneumoniae Outer Membrane Protein A Is Required to Prevent the Activation of Airway Epithelial Cells*

    PubMed Central

    March, Catalina; Moranta, David; Regueiro, Verónica; Llobet, Enrique; Tomás, Anna; Garmendia, Junkal; Bengoechea, José A.

    2011-01-01

    Outer membrane protein A (OmpA) is a class of proteins highly conserved among the Enterobacteriaceae family and throughout evolution. Klebsiella pneumoniae is a capsulated Gram-negative pathogen. It is an important cause of community-acquired and nosocomial pneumonia. Evidence indicates that K. pneumoniae infections are characterized by a lack of an early inflammatory response. Data from our laboratory indicate that K. pneumoniae CPS helps to suppress the host inflammatory response. However, it is unknown whether K. pneumoniae employs additional factors to modulate host inflammatory responses. Here, we report that K. pneumoniae OmpA is important for immune evasion in vitro and in vivo. Infection of A549 and normal human bronchial cells with 52OmpA2, an ompA mutant, increased the levels of IL-8. 52145-ΔwcaK2ompA, which does not express CPS and ompA, induced the highest levels of IL-8. Both mutants could be complemented. In vivo, 52OmpA2 induced higher levels of tnfα, kc, and il6 than the wild type. ompA mutants activated NF-κB, and the phosphorylation of p38, p44/42, and JNK MAPKs and IL-8 induction was via NF-κB-dependent and p38- and p44/42-dependent pathways. 52OmpA2 engaged TLR2 and -4 to activate NF-κB, whereas 52145-ΔwcaK2ompA activated not only TLR2 and TLR4 but also NOD1. Finally, we demonstrate that the ompA mutant is attenuated in the pneumonia mouse model. The results of this study indicate that K. pneumoniae OmpA contributes to attenuate airway cell responses. This may facilitate pathogen survival in the hostile environment of the lung. PMID:21278256

  5. First Report of Chronic Pulmonary Infection by KPC-3-Producing and Colistin-Resistant Klebsiella pneumoniae Sequence Type 258 (ST258) in an Adult Patient with Cystic Fibrosis

    PubMed Central

    Delfino, Emanuele; Del Bono, Valerio; Coppo, Erika; Marchese, Anna; Manno, Graziana; Morelli, Patrizia; Minicucci, Laura; Viscoli, Claudio

    2015-01-01

    The spread of Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae continues to increase, and the possible development of KPC-producing K. pneumoniae infections in cystic fibrosis (CF) patients is a matter of concern. Here, we describe the establishment of a chronic lung infection due to a colistin-resistant KPC-producing K. pneumoniae isolate in an Italian CF patient. PMID:25653395

  6. Phylogenetic groups among Klebsiella pneumoniae isolates from Brazil: relationship with antimicrobial resistance and origin.

    PubMed

    de Melo, Maíra Espíndola Silva; Cabral, Adriane Borges; Maciel, Maria Amélia Vieira; da Silveira, Vera Magalhães; de Souza Lopes, Ana Catarina

    2011-05-01

    The objectives of this study were to determine the distribution of phylogenetic groups among Klebsiella pneumoniae isolates from Recife, Brazil and to assess the relationship between the groups and the isolation sites and resistance profile. Ninety four isolates of K. pneumoniae from hospital or community infections and from normal microbiota were analyzed by gyrA PCR-RFLP, antibiotic susceptibility, and adonitol fermentation. The results revealed the distinction of three phylogenetic groups, as it has also been reported in Europe, showing that these clusters are highly conserved within K. pneumoniae. Group KpI was dominantly represented by hospital and community isolates while groups KpII and KpIII displayed mainly normal microbiota isolates. The resistance to third generation cephalosporins, aztreonam, imipenem, amoxicillin/clavulanic acid, and streptomycin was only observed in KpI. The percentage of resistance was higher in KpI, followed by KpII and KpIII. The differences in the distribution of K. pneumoniae phylogenetic groups observed in this study suggest distinctive clinical and epidemiological characteristics among the three groups, which is important to understand the epidemiology of infections caused by this organism. This is the first study in Brazil on K. pneumoniae isolates from normal microbiota and community infections regarding the distribution of phylogenetic groups based on the gyrA gene.

  7. The metabolic flux regulation of Klebsiella pneumoniae based on quorum sensing system

    PubMed Central

    Sun, Shujing; Zhang, Haiyang; Lu, Shuyi; Lai, Chunfen; Liu, Huijun; Zhu, Hu

    2016-01-01

    Quorum-sensing (QS) systems exist universally in bacteria to regulate multiple biological functions. Klebsiella pneumoniae, an industrially important bacterium that produces bio-based chemicals such as 2,3-butanediol and acetoin, can secrete a furanosyl borate diester (AI-2) as the signalling molecule mediating a QS system, which plays a key regulatory role in the biosynthesis of secondary metabolites. In this study, the molecular regulation and metabolic functions of a QS system in K. pneumoniae were investigated. The results showed that after the disruption of AI-2-mediated QS by the knockout of luxS, the production of acetoin, ethanol and acetic acid were relatively lower in the K. pneumoniae mutant than in the wild type bacteria. However, 2,3-butanediol production was increased by 23.8% and reached 54.93 g/L. The observed enhancement may be attributed to the improvement of the catalytic activity of 2,3-butanediol dehydrogenase (BDH) in transforming acetoin to 2,3-butanediol. This possibility is consistent with the RT-PCR-verified increase in the transcriptional level of budC, which encodes BDH. These results also demonstrated that the physiological metabolism of K. pneumoniae was adversely affected by a QS system. This effect was reversed through the addition of synthetic AI-2. This study provides the basis for a QS-modulated metabolic engineering study of K. pneumoniae. PMID:27924940

  8. Role of hydrogen generation by Klebsiella pneumoniae in the oral cavity.

    PubMed

    Kanazuru, Tomoko; Sato, Eisuke F; Nagata, Kumiko; Matsui, Hiroshi; Watanabe, Kunihiko; Kasahara, Emiko; Jikumaru, Mika; Inoue, June; Inoue, Masayasu

    2010-12-01

    Some gastrointestinal bacteria synthesize hydrogen (H(2)) by fermentation. Despite the presence of bactericidal factors in human saliva, a large number of bacteria also live in the oral cavity. It has never been shown that oral bacteria also produce H(2) or what role H(2) might play in the oral cavity. It was found that a significant amount of H(2) is synthesized in the oral cavity of healthy human subjects, and that its generation is enhanced by the presence of glucose but inhibited by either teeth brushing or sterilization with povidone iodine. These observations suggest the presence of H(2)-generating bacteria in the oral cavity. The screening of commensal bacteria in the oral cavity revealed that a variety of anaerobic bacteria generate H(2). Among them, Klebsiella pneumoniae (K. pneumoniae) generated significantly large amounts of H(2) in the presence of glucose. Biochemical analysis revealed that various proteins in K. pneumoniae are carbonylated under standard culture conditions, and that oxidative stress induced by the presence of Fe(++) and H(2)O(2) increases the number of carbonylated proteins, particularly when their hydrogenase activity is inhibited by KCN. Inhibition of H(2) generation markedly suppresses the growth of K. pneumoniae. These observations suggest that H(2) generation and/or the reduction of oxidative stress is important for the survival and growth of K. pneumoniae in the oral cavity.

  9. Molecular epidemiology of aminoglycosides resistance on Klebsiella pneumonia in a hospital in China.

    PubMed

    Liang, Caiqian; Xing, Bangrong; Yang, Xiaoyan; Fu, Yongmei; Feng, Yaqun; Zhang, Yongbiao

    2015-01-01

    To investigate the molecular epidemiology of aminoglycosides resistance among Klebsiella pneumonia in hospitals in China, the antibiotics resistance and the possession of extended-spectrum β-lactamases (ESBLs) from 162 isolates were examined using Kirby-Bauer disk diffusion and PCR sequencing. Overall, 47.5% (77/162) of strains showed an ESBL phenotype. According to antibiotics resistance, ESBLs-positive K. pneumoniae showed significantly higher resistance to most antibiotics than ESBLs-negative strains (P<0.05). Moreover, 162 strains harboured aminoglycoside-modifying enzymes genes (AMEs) including aac (3)-II (n = 49), aac (6')-Ib (n = 32), ant (3")-I (n = 22) and ant (2")-I (n = 7). Overall, 11.1% (18/162) and 6.2% (10/162) of isolates carried 16S rRNA methylase genes (armA and rmtB), in which the aminoglycoside MIC was more than 256 μg/ml. In conclusion, our study characterised aminoglycosides resistance among K. pneumoniae strains in China hospitals and revealed antibiotic resistance and the increased presence of AMEs and 16S rRNA methylase genes in K. pneumonia, enabling the prevalence of aminoglycosides resistance of K. pneumoniae to be tracked from patients.

  10. Genomic Definition of Hypervirulent and Multidrug-Resistant Klebsiella pneumoniae Clonal Groups

    PubMed Central

    Bialek-Davenet, Suzanne; Criscuolo, Alexis; Ailloud, Florent; Passet, Virginie; Jones, Louis; Delannoy-Vieillard, Anne-Sophie; Garin, Benoit; Le Hello, Simon; Arlet, Guillaume; Nicolas-Chanoine, Marie-Hélène; Decré, Dominique

    2014-01-01

    Multidrug-resistant and highly virulent Klebsiella pneumoniae isolates are emerging, but the clonal groups (CGs) corresponding to these high-risk strains have remained imprecisely defined. We aimed to identify K. pneumoniae CGs on the basis of genome-wide sequence variation and to provide a simple bioinformatics tool to extract virulence and resistance gene data from genomic data. We sequenced 48 K. pneumoniae isolates, mostly of serotypes K1 and K2, and compared the genomes with 119 publicly available genomes. A total of 694 highly conserved genes were included in a core-genome multilocus sequence typing scheme, and cluster analysis of the data enabled precise definition of globally distributed hypervirulent and multidrug-resistant CGs. In addition, we created a freely accessible database, BIGSdb-Kp, to enable rapid extraction of medically and epidemiologically relevant information from genomic sequences of K. pneumoniae. Although drug-resistant and virulent K. pneumoniae populations were largely nonoverlapping, isolates with combined virulence and resistance features were detected. PMID:25341126

  11. Phenotypic and genotypic characterization of Klebsiella pneumoniae isolates recovered from nonhuman primates.

    PubMed

    Soto, Esteban; LaMon, Virginia; Griffin, Matt; Keirstead, Natalie; Beierschmitt, Amy; Palmour, Roberta

    2012-07-01

    Klebsiella pneumoniae is a zoonotic, Gram-negative member of the family Enterobacteriaceae and is the causative agent of nosocomial septicemic, pneumonic, and urinary tract infections. Recently, pathogenic strains of K. pneumoniae sharing a hypermucoviscosity (HMV) phenotype have been attributed to multisystemic abscessation in both human and nonhuman primates. Although K. pneumoniae is a well-recognized zoonotic agent, there is a lack of general information including adequate diagnostic methods or treatments for nonhuman primates. In an effort to increase the body of knowledge of this enigmatic pathogen, K. pneumoniae isolates from African green monkeys (Chlorocebus aethiops sabaeus) on the island of St. Kitts, West Indies were genotypically and phenotypically characterized. Genetic fingerprints generated by PCR-mediated genomic fingerprinting, phenotypic characterization, and antimicrobial susceptibility all identified a high degree of similarity between the HMV and non-HMV K. pneumoniae isolates. The results obtained from this work will help establish a baseline for the development of efficacious diagnostic methods and treatment strategies for both human and nonhuman primates.

  12. Molecular epidemiology of extensively drug-resistant Klebsiella pneumoniae outbreak in Wenzhou, Southern China.

    PubMed

    Du, Jia; Cao, Jianming; Shen, Lizhen; Bi, Wenzi; Zhang, Xiaoxiao; Liu, Haiyang; Lu, Hong; Zhou, Tieli

    2016-10-01

    The emergence of extensively drug-resistant (XDR) Klebsiella pneumoniae has become a major challenge worldwide. In this study, we characterized the phenotypes and genetic features of nine XDR K. pneumoniae isolates from an integrated hospital in Zhejiang province, China, from September to October 2014. These XDR K. pneumoniae possessed at least five resistance determinants which contribute to highly resistant to β-lactam, β-lactam/inhibitor combinations, aminoglycosides, quinolones, carbapenems, chloroamphenicol and fosfomycin. All isolates carried blaKPC-2, blaCTX-M-9, blaSHV-11 and rmtB, and several isolates also harboured blaTEM-1 and qnrS. Southern blot experiments confirmed that blaKPC-2, rmtB and blaCTX-M-9 were located on the same ~54.2 kb plasmid. Conjugative plasmids were obtained from all K. pneumoniae isolates, further proving the transferable characteristic of the resistance determinants. The OmpK36 sequences showed various deletions and insertions that indicated additional amino acid residues and a deleted phenotype of OmpK36. PFGE demonstrated that all the isolates belonged to the same genotype. Multilocus sequence typing was concordant with PFGE results and revealed that ST11 was the most predominant clone. Our study revealed a high incidence and endemic spread of XDR K. pneumoniae in the hospital. Thus, effective infection control measures should be adopted to monitor and control the spread of multidrug-resistant isolates.

  13. Multicentre investigation of carbapenemase-producing Escherichia coli and Klebsiella pneumoniae in German hospitals.

    PubMed

    Kaase, Martin; Schimanski, Sven; Schiller, Reinhold; Beyreiß, Bettina; Thürmer, Alexander; Steinmann, Jörg; Kempf, Volkhard A; Hess, Christina; Sobottka, Ingo; Fenner, Ines; Ziesing, Stefan; Burckhardt, Irene; von Müller, Lutz; Hamprecht, Axel; Tammer, Ina; Wantia, Nina; Becker, Karsten; Holzmann, Thomas; Furitsch, Martina; Volmer, Gabriele; Gatermann, Sören G

    2016-09-01

    Aim of this study was to determine the incidence and molecular epidemiology of carbapenemase-producing Escherichia coli and Klebsiella pneumoniae in Germany. E. coli and K. pneumoniae isolates from clinical samples which were non-susceptible to carbapenems were collected in laboratories serving 20 hospitals throughout Germany from November 2013 to April 2014. The isolates were tested for the presence of carbapenemases by PCR and phenotypic methods and typed by multilocus sequence typing. Risk factors including a previous hospitalization abroad were analysed. Carbapenemases were detected in 24 isolates from 22 patients out of 464,514 admissions. Carbapenemases included OXA-48 (n=14), KPC-2 (n=8) and NDM-1 (n=2). Except for two K. pneumoniae isolates with ST101, all OXA-48 producing strains belonged to different clones. In contrast, half of KPC-2 producing K. pneumoniae were of ST258 and both NDM-1 producing strains were of ST11. Compared to carbapenem-susceptible controls, patients with carbapenemase-producing strains differed by a significantly higher proportion of males, a higher proportion of isolates from wound samples and a more frequent previous stay abroad in univariate analysis. This multicentre study demonstrated an incidence of carbapenemase-producing E. coli and K. pneumoniae from clinical samples in Germany of 0.047 cases per 1000 admissions. OXA-48 was more frequent than KPC-2 and NDM-1 and showed a multiclonal background.

  14. Genomic definition of hypervirulent and multidrug-resistant Klebsiella pneumoniae clonal groups.

    PubMed

    Bialek-Davenet, Suzanne; Criscuolo, Alexis; Ailloud, Florent; Passet, Virginie; Jones, Louis; Delannoy-Vieillard, Anne-Sophie; Garin, Benoit; Le Hello, Simon; Arlet, Guillaume; Nicolas-Chanoine, Marie-Hélène; Decré, Dominique; Brisse, Sylvain

    2014-11-01

    Multidrug-resistant and highly virulent Klebsiella pneumoniae isolates are emerging, but the clonal groups (CGs) corresponding to these high-risk strains have remained imprecisely defined. We aimed to identify K. pneumoniae CGs on the basis of genome-wide sequence variation and to provide a simple bioinformatics tool to extract virulence and resistance gene data from genomic data. We sequenced 48 K. pneumoniae isolates, mostly of serotypes K1 and K2, and compared the genomes with 119 publicly available genomes. A total of 694 highly conserved genes were included in a core-genome multilocus sequence typing scheme, and cluster analysis of the data enabled precise definition of globally distributed hypervirulent and multidrug-resistant CGs. In addition, we created a freely accessible database, BIGSdb-Kp, to enable rapid extraction of medically and epidemiologically relevant information from genomic sequences of K. pneumoniae. Although drug-resistant and virulent K. pneumoniae populations were largely nonoverlapping, isolates with combined virulence and resistance features were detected.

  15. Elucidation of the RamA Regulon in Klebsiella pneumoniae Reveals a Role in LPS Regulation

    PubMed Central

    De Majumdar, Shyamasree; Yu, Jing; Fookes, Maria; McAteer, Sean P.; Llobet, Enrique; Finn, Sarah; Spence, Shaun; Monaghan, Avril; Kissenpfennig, Adrien; Ingram, Rebecca J.; Bengoechea, José; Gally, David L.; Fanning, Séamus; Elborn, Joseph S.; Schneiders, Thamarai

    2015-01-01

    Klebsiella pneumoniae is a significant human pathogen, in part due to high rates of multidrug resistance. RamA is an intrinsic regulator in K. pneumoniae established to be important for the bacterial response to antimicrobial challenge; however, little is known about its possible wider regulatory role in this organism during infection. In this work, we demonstrate that RamA is a global transcriptional regulator that significantly perturbs the transcriptional landscape of K. pneumoniae, resulting in altered microbe-drug or microbe-host response. This is largely due to the direct regulation of 68 genes associated with a myriad of cellular functions. Importantly, RamA directly binds and activates the lpxC, lpxL-2 and lpxO genes associated with lipid A biosynthesis, thus resulting in modifications within the lipid A moiety of the lipopolysaccharide. RamA-mediated alterations decrease susceptibility to colistin E, polymyxin B and human cationic antimicrobial peptide LL-37. Increased RamA levels reduce K. pneumoniae adhesion and uptake into macrophages, which is supported by in vivo infection studies, that demonstrate increased systemic dissemination of ramA overexpressing K. pneumoniae. These data establish that RamA-mediated regulation directly perturbs microbial surface properties, including lipid A biosynthesis, which facilitate evasion from the innate host response. This highlights RamA as a global regulator that confers pathoadaptive phenotypes with implications for our understanding of the pathogenesis of Enterobacter, Salmonella and Citrobacter spp. that express orthologous RamA proteins. PMID:25633080

  16. Two distinct sensing pathways allow recognition of Klebsiella pneumoniae by Dictyostelium amoebae.

    PubMed

    Lima, Wanessa C; Balestrino, Damien; Forestier, Christiane; Cosson, Pierre

    2014-03-01

    Recognition of bacteria by metazoans is mediated by receptors that recognize different types of microorganisms and elicit specific cellular responses. The soil amoebae Dictyostelium discoideum feeds upon a variable mixture of environmental bacteria, and it is expected to recognize and adapt to various food sources. To date, however, no bacteria-sensing mechanisms have been described. In this study, we isolated a Dictyostelium mutant (fspA KO) unable to grow in the presence of non-capsulated Klebsiella pneumoniae bacteria, but growing as efficiently as wild-type cells in the presence of other bacteria, such as Bacillus subtilis. fspA KO cells were also unable to respond to K. pneumoniae and more specifically to bacterially secreted folate in a chemokinetic assay, while they responded readily to B. subtilis. Remarkably, both WT and fspA KO cells were able to grow in the presence of capsulated LM21 K. pneumoniae, and responded to purified capsule, indicating that capsule recognition may represent an alternative, FspA-independent mechanism for K. pneumoniae sensing. When LM21 capsule synthesis genes were deleted, growth and chemokinetic response were lost for fspA KO cells, but not for WT cells. Altogether, these results indicate that Dictyostelium amoebae use specific recognition mechanisms to respond to different K. pneumoniae elements.

  17. Inactivation of the virulence factors from 2,3-butanediol-producing Klebsiella pneumoniae.

    PubMed

    Huynh, Duyen Thi Ngoc; Kim, Ah-Young; Seol, In-Hye; Jung, Samuel; Lim, Min-Cheol; Lee, Jeong-A; Jo, Mi-Rae; Choi, Soo-Jin; Kim, Borim; Lee, Jinwon; Kim, Wooki; Kim, Young-Rok

    2015-11-01

    The microbiological production of 2,3-butanediol (2,3-BDO) has attracted considerable attention as an alternative way to produce high-value chemicals from renewable sources. Among the number of 2,3-BDO-producing microorganisms, Klebsiella pneumoniae has been studied most extensively and is known to produce large quantity of 2,3-BDO from a range of substrates. On the other hand, the pathogenic characteristics of the bacteria have limited its industrial applications. In this study, two major virulence traits, outer core LPS and fimbriae, were removed through homologous recombination from 2,3-BDO-producing K. pneumoniae 2242 to expand its uses to the industrial scale. The K. pneumoniae 2242 ∆wabG mutant strain was found to have an impaired capsule, which significantly reduced its ability to bind to the mucous layer and evade the phagocytic activity of macrophage. The association with the human ileocecal epithelial cell, HCT-8, and the bladder epithelial cell, T-24, was also reduced dramatically in the K. pneumoniae 2242 ∆fimA mutant strain that was devoid of fimbriae. However, the growth rate and production yield for 2,3-BDO were unaffected. The K. pneumoniae strains developed in this study, which are devoid of the major virulence factors, have a high potential for the efficient and sustainable production of 2,3-BDO.

  18. Rapid glia expression and release of proinflammatory cytokines in experimental Klebsiella pneumoniae meningoencephalitis.

    PubMed

    Wen, Li-Li; Chiu, Chien-Tsai; Huang, Ya-Ni; Chang, Che-Feng; Wang, Jia-Yi

    2007-05-01

    The host immune/inflammatory response following CNS infection by Klebsiella pneumoniae remains poorly understood. Using a rat model of K. pneumoniae meningoencephalitis, we investigated the temporal profiles of brain proinflammatory cytokines and their cellular sources. Leukocyte counts significantly increased in cerebrospinal fluid (CSF) at 2 h after K. pneumoniae inoculation into the rat brain but were still much lower than blood leukocyte counts. However, concentrations of tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), and IL-6 in CSF were much higher than the simultaneously collected serum levels. The rapid increase in brain expression of these cytokines at the messenger RNA (mRNA) and protein levels occurred earlier than the onset of leukocytosis. Double immunofluorescence staining revealed the presence of TNF-alpha, IL-1beta, and IL-6 in astrocytes and microglia. Exposure of primary culture of glial cells to K. pneumoniae also resulted in time-dependent increases in the concentration of these cytokines in the culture media. Taken together, our results suggest that glial cells are an important early source of proinflammatory cytokines during K. pneumonia infection of CNS.

  19. Klebsiella pneumoniae alleviates influenza-induced acute lung injury via limiting NK cell expansion.

    PubMed

    Wang, Jian; Li, Fengqi; Sun, Rui; Gao, Xiang; Wei, Haiming; Tian, Zhigang

    2014-08-01

    A protective effect induced by bacterial preinfection upon a subsequent lethal influenza virus infection has been observed, but the underlying immune mechanisms have not yet been fully elucidated. In this study, we used a mouse model of Klebsiella pneumoniae preinfection to gain insight into how bacterial preinfection influences the subsequent lethal influenza virus infection. We found that K. pneumoniae preinfection significantly attenuated lung immune injury and decreased mortality during influenza virus infection, but K. pneumoniae-specific immunity was not involved in this cross-protection against influenza virus. K. pneumoniae preinfection limited NK cell expansion, which was involved in influenza-induced immune injury and death. Furthermore, K. pneumoniae preinfection could not control NK cell expansion and death during influenza virus infection in Rag1(-/-) mice, but adoptive transfer of T cells from wild-type mice was able to restore this protective effect. Our data suggest that the adaptive immune response activated by bacterial infection limits the excessive innate immune response induced by a subsequent influenza infection, ultimately protecting mice from death. Copyright © 2014 by The American Association of Immunologists, Inc.

  20. An observational study of phagocytes and Klebsiella pneumoniae relationships: different behaviors.

    PubMed

    Maisonneuve, Elodie; Cateau, Estelle; Delouche, Marion; Quellard, Nathalie; Rodier, Marie-Helene

    2017-01-10

    Klebsiella pneumoniae is a bacterium that can be in relation with free living amoebae like Acanthamoeba castellanii in natural environments such as soil and water. This pathogen, which is responsible for community-acquired pneumonia and for nosocomial infections, also has interactions with host defense mechanisms like macrophages. As it has been shown that A. castellanii shares some traits with macrophages, in particular the ability to phagocyte bacteria, we have studied the uptake and the fate of the bacteria after contact with the two phagocytic cells. In our conditions, K. pneumoniae growth was increased in coculture in presence of A. castellanii or Thp-1 macrophagic cells and bacterial development was also increased by A. castellanii supernatant. In addition, we showed that the presence of the bacteria had a negative effect on the macrophages whereas it does not affect amoeba viability. Using gentamicin, which kills bacteria outside cells, we showed that only macrophages were able to internalize K. pneumoniae. This result was confirmed by electron microscopy. We have consequently reported some differences in bacterial uptake and internalization between a free living amoeba and macrophagic cells, highlighting the fact that results obtained with this amoebal model should not be extrapolated to the relationships between K. pneumoniae and macrophages.

  1. Two distinct sensing pathways allow recognition of Klebsiella pneumoniae by Dictyostelium amoebae

    PubMed Central

    Lima, Wanessa C; Balestrino, Damien; Forestier, Christiane; Cosson, Pierre

    2013-01-01

    Summary Recognition of bacteria by metazoans is mediated by receptors that recognize different types of microorganisms and elicit specific cellular responses. The soil amoebae Dictyostelium discoideum feeds upon a variable mixture of environmental bacteria, and it is expected to recognize and adapt to various food sources. To date, however, no bacteria-sensing mechanisms have been described. In this study, we isolated a Dictyostelium mutant (fspA KO) unable to grow in the presence of non-capsulated Klebsiella pneumoniae bacteria, but growing as efficiently as wild-type cells in the presence of other bacteria, such as Bacillus subtilis. fspA KO cells were also unable to respond to K. pneumoniae and more specifically to bacterially secreted folate in a chemokinetic assay, while they responded readily to B. subtilis. Remarkably, both WT and fspA KO cells were able to grow in the presence of capsulated LM21 K. pneumoniae, and responded to purified capsule, indicating that capsule recognition may represent an alternative, FspA-independent mechanism for K. pneumoniae sensing. When LM21 capsule synthesis genes were deleted, growth and chemokinetic response were lost for fspA KO cells, but not for WT cells. Altogether, these results indicate that Dictyostelium amoebae use specific recognition mechanisms to respond to different K. pneumoniae elements. PMID:24128258

  2. Klebsiella Pneumoniae Liver Abscess: a Case Report and Review of Literature

    PubMed Central

    Williams, George; Akbar, Hina; Khan, Muhammad Ali; Kadaria, Dipen

    2017-01-01

    Klebsiella pneumoniae (K.pneumoniae) is a known cause of pyogenic liver abscess (PLA) in the absence of hepatobiliary disease. In settings of hepatic infection, it has also been known to cause disseminated infections including meningitis and endopthalmitis. Several groups of patients are particularly susceptible to infection, including patients with diabetes mellitus, those from Southeast Asia and those with the preexisting hepatobiliary disease. We present a case of K.pneumoniae PLA with bacteremia. A 39-year-old Vietnamese male with no previous medical history who presented with complaints of abdominal pain, nausea, vomiting, diarrhea and fever. A computed tomography (CT) of the abdomen showed a large complex mass in the right lobe of the liver with multiple septations. Over course of hospitalization, the patient developed acute respiratory failure and was monitored in medical intensive care unit (MICU). Blood cultures grew K. pneumonia. The patient was treated with intravenous ceftriaxone and the abscess was drained by interventional radiology. After appropriate management, he progressed well during his hospital course and was eventually discharged from the hospital. K. pneumonia PLA had previously been an endemic disease in Southeast Asia, however, with a highly mobile patient population, it is now seen throughout the world and should be in the differential of patients who present with solitary liver mass in the setting of sepsis. PMID:28191374

  3. Emergence of Klebsiella pneumoniae clinical isolates producing KPC-2 carbapenemase in Cuba.

    PubMed

    Quiñones, D; Hart, M; Espinosa, F; Garcia, S; Carmona, Y; Ghosh, S; Urushibara, N; Kawaguchiya, M; Kobayashi, N

    2014-07-01

    The emergence of Klebsiella pneumoniae producing carbapenemase (KPC) has now become a global concern. As a part of a nationwide multicentre surveillance study in Cuba, three K. pneumoniae clinical isolates resistant to carbapenems were detected for a 1-month period (September to October 2011). PCR and sequence analysis revealed that the three strains harboured bla KPC-2. They showed resistance or intermediate susceptibility to expanded-spectrum cephalosporins, other β-lactams, a β-lactam/β-lactamase inhibitor combination, and gentamicin. Two strains were susceptible only to colistin, whereas the other strain showing colistin resistance was susceptible to fluoroquinolones. These bla KPC -2-positive K. pneumoniae strains were classified into ST1271 (CC29), a novel clone harbouring bla KPC -2, and were revealed to be genetically identical by PCR-based DNA fingerprinting. The three patients infected with the KPC-producing K. pneumoniae had common risk factors, and had no overseas travel experience outside Cuba, suggesting local acquisition of the resistant pathogen. This is the first report of a KPC-producing K. pneumoniae in Cuba. Although detection of KPC in Enterobacteriaceae is still rare in Cuba, our finding indicated that KPC-producing bacteria are a global concern and highlighted the need to identify these microorganisms in clinical laboratories.

  4. Emergence of Klebsiella pneumoniae clinical isolates producing KPC-2 carbapenemase in Cuba

    PubMed Central

    Quiñones, D; Hart, M; Espinosa, F; Garcia, S; Carmona, Y; Ghosh, S; Urushibara, N; Kawaguchiya, M; Kobayashi, N

    2014-01-01

    The emergence of Klebsiella pneumoniae producing carbapenemase (KPC) has now become a global concern. As a part of a nationwide multicentre surveillance study in Cuba, three K. pneumoniae clinical isolates resistant to carbapenems were detected for a 1-month period (September to October 2011). PCR and sequence analysis revealed that the three strains harboured blaKPC-2. They showed resistance or intermediate susceptibility to expanded-spectrum cephalosporins, other β-lactams, a β-lactam/β-lactamase inhibitor combination, and gentamicin. Two strains were susceptible only to colistin, whereas the other strain showing colistin resistance was susceptible to fluoroquinolones. These blaKPC-2-positive K. pneumoniae strains were classified into ST1271 (CC29), a novel clone harbouring blaKPC-2, and were revealed to be genetically identical by PCR-based DNA fingerprinting. The three patients infected with the KPC-producing K. pneumoniae had common risk factors, and had no overseas travel experience outside Cuba, suggesting local acquisition of the resistant pathogen. This is the first report of a KPC-producing K. pneumoniae in Cuba. Although detection of KPC in Enterobacteriaceae is still rare in Cuba, our finding indicated that KPC-producing bacteria are a global concern and highlighted the need to identify these microorganisms in clinical laboratories. PMID:25356357

  5. In silico identification of potential virulence genes in 1,3-propanediol producer Klebsiella pneumonia.

    PubMed

    Gao, L R; Jiang, X; Fu, S L; Gong, H

    2014-11-10

    The pathogenic characteristics of Klebsiella pneumoniae could pose security risks for industrial applications. In this study, the existence and distribution of 2457 known virulence genes (VFs) in 9 strains of K. pneumoniae were systematically analyzed by high-throughput in silico methods. We found different numbers and types of VFs in 9 K. pneumoniae strains using database sequences. Some VFs in the database were highly homologous with the corresponding genes in K. pneumoniae genomes. Four large fragments with contiguous potential virulence genes named VF1, VF2, VF3 and VF4 were identified. VF1 and VF2 were found in all 9 sequenced strains and the 1,3-propanediol-producing strain KG1. When the VF2 fragment was knocked out in KG1, cell growth and 1,3-propanediol production in the mutant were nearly the same as in KG1. Consequently the resulting information by in silico methods is useful for identifying potential virulence genes of K. pneumoniae used for 1,3-propanediol production.

  6. Clonal replacement of epidemic KPC-producing Klebsiella pneumoniae in a hospital in China.

    PubMed

    Liang, Yuying; Yin, Xiuyun; Zeng, Lijun; Chen, Shuiping

    2017-05-23

    Klebsiella pneumoniae is a frequent nosocomial pathogen causing difficult-to-treat infections worldwide. The prevalence of Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae (KPC-KP) is increasing in China. The aim of this study was to investigate the molecular epidemiology of KPC-KP in a nosocomial outbreak. Fifty-four KPC-KP isolates were consecutively collected between November 2013 and August 2014 during a KPC-KP outbreak in a tertiary care hospital in Beijing, China. Antimicrobial susceptibility was determined by agar dilution. Carbapenemase, extended-spectrum β-lactamase, 16S rRNA methylase, AmpC β-lactamase, and plasmid-mediated quinolone resistance determinants were detected by PCR amplification. The genetic relatedness of isolates was analyzed by pulsed-field gel electrophoresis and multi-locus sequence typing. All isolates belonged to ST11 except one isolate which was identified as a new sequence type (ST2040). PFGE profile of genomic DNA revealed seven clusters, of which cluster A and C dominated the KPC-KP outbreak and cluster A was replaced by cluster C during the outbreak. PFGE of genomic DNA, S1-PFGE of plasmids, replicon typing, and drug resistant characteristics showed that clonal spread occurred during the outbreak. When compared with isolates within cluster A, all isolates in cluster C harbored rmtB and showed higher level of resistance to cefepime, amikacin, tobramycin, and tigecycline. We reported a nosocomial outbreak of KPC-KP with clonal replacement and a new sequence type (ST2040) of KP. High degree of awareness and surveillance of KPC-KP should be given to avoid potential outbreaks, especially in ICU wards.

  7. [Protection and mechanism of Fagopyrum cymosum on lung injury in rats with Klebsiella pneumonia].

    PubMed

    Dong, Liu-Yi; Wang, Chun-Yan; Wu, Chang-Qing; Jiang, Qin; Zhang, Zhi-Fen

    2012-04-01

    To study the mechanism of protective effect of Fagopyrum cymosum on lung injury induced by Klebsiella pneumonia in rats. The model of rats with Klebsiella pneumonia was established. The male SD rats were randomly divided into control group, model group, Fagopyrum cymosum (6, 3, 1.5 g/kg) three groups, levofloxacin (25 mg/kg) group. The pathological change of lung was observed. The content of IL-1beta, IL-6, IL-8, TNF-alpha, ICAM-1, INF-gamma in serum were measured by radioimmunoassay and Elisa. TNF-alpha, ICAM-1, NF-kappaB p65 protein expressions were measured by immunohistochemistry. MIP-2mRNA expression was detected by in situ hybridization. The rats of model group had obvious lung injury, but those of Fagopyrum cymosum and levofloxacin groups had less injury. The contents of IL-1beta, IL-6, IL-,8, TNF-alpha, ICAM-1 and INF-gamma in serum and the expressions of TNF-a, ICAM-1, NF-kappaB p65 and MIP--2mRNA of model group were significantly higher than those of the control group (P < 0.05 or P < 0.01), while the indexes of Fagopyrum cymosum and levofloxacin groups were significantly lower than those of model group (P < 0.05 or P < 0.01). The lung injury induced by Klebsiella pneumonia is related to TNF-alpha, ICAM-1, NF-kappaB p65 and MIP-2mRNA. To decrease the excessive expression of TNF-alpha, ICAM-1, NF-kappaB p65 and MIP-2mRNA might be the main mechanism of protective effect of Fagopyrum cymosum on lung injury.

  8. Molecular characterization of newly emerged blaKPC-2-producing Klebsiella pneumoniae in Singapore.

    PubMed

    Balm, Michelle N D; Ngan, Grace; Jureen, Roland; Lin, Raymond T P; Teo, Jeanette

    2012-02-01

    In Asia, bla(KPC) detection has been limited to East Asia and not yet seen in Southeast Asia. We report four bla(KPC-2)-containing Klebsiella pneumoniae isolates from two different hospitals in Singapore. All isolates belonged to strain type 11 (ST11) and were indistinguishable by pulsed-field gel electrophoresis (PFGE). bla(KPC-2) was located on nonconjugative plasmids and flanked by mobile genetic structures composed of a partial Tn4401 transposon and a Tn3-based transposon which previously have been described only in Chinese isolates.

  9. Molecular Characterization of Newly Emerged blaKPC-2-Producing Klebsiella pneumoniae in Singapore

    PubMed Central

    Ngan, Grace; Jureen, Roland; Lin, Raymond T. P.; Teo, Jeanette

    2012-01-01

    In Asia, blaKPC detection has been limited to East Asia and not yet seen in Southeast Asia. We report four blaKPC-2-containing Klebsiella pneumoniae isolates from two different hospitals in Singapore. All isolates belonged to strain type 11 (ST11) and were indistinguishable by pulsed-field gel electrophoresis (PFGE). blaKPC-2 was located on nonconjugative plasmids and flanked by mobile genetic structures composed of a partial Tn4401 transposon and a Tn3-based transposon which previously have been described only in Chinese isolates. PMID:22116160

  10. Outbreak of NDM-1-Producing Klebsiella pneumoniae in a Neonatal Unit in Colombia

    PubMed Central

    Olarte Escobar, Narda María; Castro-Cardozo, Betsy; Valderrama Márquez, Ismael Alberto; Garzón Aguilar, Martha Isabel; Martinez de la Barrera, Leslie; Barrero Barreto, Esther Rocio; Marquez-Ortiz, Ricaurte Alejandro; Moncada Guayazán, Maria Victoria; Vanegas Gómez, Natasha

    2013-01-01

    Six multiresistant, NDM-1-producing Klebsiella pneumoniae strains were recovered from an outbreak that affected six neonatal patients in a Colombian hospital. Molecular analysis showed that all of the isolates harbored the blaNDM-1, qnrA, and intI1 genes and were clonally related. Multilocus sequence typing showed that the isolates belonged to a new sequence type (ST1043) that was different from the sequence types that had previously been reported. This is the first report of NDM-1-producing isolates in South America. PMID:23357776

  11. Emergence of Carbapenem-Resistant Serotype K1 Hypervirulent Klebsiella pneumoniae Strains in China

    PubMed Central

    Zhang, Rong; Lin, Dachuan; Chan, Edward Wai-chi; Gu, Danxia

    2015-01-01

    We report the emergence of five carbapenem-resistant K1 hypervirulent Klebsiella pneumoniae (hvKP) strains which caused fatal infections in hospital patients in Zhejiang Province, China, upon entry through surgical wounds. Genotyping results revealed the existence of three genetically related strains which exhibited a new sequence type, ST1797, and revealed that all strains harbored the magA and wcaG virulence genes and a plasmid-borne blaKPC-2 gene. These findings indicate that K1 hvKP is simultaneously hypervirulent, multidrug resistant, and transmissible. PMID:26574010

  12. Novel plasmid-encoded class C beta-lactamase (MOX-2) in Klebsiella pneumoniae from Greece.

    PubMed

    Raskine, Laurent; Borrel, Isabelle; Barnaud, Guilène; Boyer, Sophie; Hanau-Berçot, Béatrice; Gravisse, Jérome; Labia, Roger; Arlet, Guillaume; Sanson-Le-Pors, Marie-José

    2002-07-01

    Klebsiella pneumoniae KOL, a clinical strain resistant to various beta-lactams, was isolated from the stools of a patient from Greece. This strain harbored a new pI 9.1 plasmid-mediated AmpC beta-lactamase with unusually high levels of hydrolytic activity for cefoxitin and cefotetan that we named MOX-2. Sequencing of bla(MOX-2) revealed 93.2, 92.9, 92.7, and 73.1% identities with the deduced amino acid sequences of CMY-8, MOX-1, CMY-1, and the AmpC beta-lactamase of Aeromonas sobria, respectively.

  13. Synthesis and characterization of bactericidal silver nanoparticles using cultural filtrate of simulated microgravity grown Klebsiella pneumoniae.

    PubMed

    Kalpana, Duraisamy; Lee, Yang Soo

    2013-03-05

    Silver nanoparticles were synthesized by biological method using cultural filtrate of Klebsiella pneumoniae cultured under simulated microgravity and silver nitrate solution as precursor. The nanoparticles exhibited typical plasmon absorption maximum of silver nanoparticles between 405 and 407 nm. Spherical silver nanoparticles were found to have size between 15 and 37 nm by TEM analysis. XRD pattern corresponding to planes (111), (200), (220) (311) revealed the crystalline nature of the biosynthesized silver nanoparticles. FTIR spectrum proposed stabilization of silver nanoparticles by the protein molecules present in the cultural filtrate. The silver nanoparticles exhibited high bactericidal activity against Salmonella enterica, Escherichia coli and moderate bactericidal activity against Streptococcus pyogenes.

  14. Emergence of Carbapenem-Resistant Serotype K1 Hypervirulent Klebsiella pneumoniae Strains in China.

    PubMed

    Zhang, Rong; Lin, Dachuan; Chan, Edward Wai-Chi; Gu, Danxia; Chen, Gong-Xiang; Chen, Sheng

    2015-11-16

    We report the emergence of five carbapenem-resistant K1 hypervirulent Klebsiella pneumoniae (hvKP) strains which caused fatal infections in hospital patients in Zhejiang Province, China, upon entry through surgical wounds. Genotyping results revealed the existence of three genetically related strains which exhibited a new sequence type, ST1797, and revealed that all strains harbored the magA and wcaG virulence genes and a plasmid-borne bla(KPC-2) gene. These findings indicate that K1 hvKP is simultaneously hypervirulent, multidrug resistant, and transmissible.

  15. A Neonatal Septic Arthritis Case Caused by Klebsiella pneumoniae: A Case Report

    PubMed Central

    Ozsari, Tamer; Ozdemir, Özmert M.A; Kiliç, Ilknur

    2016-01-01

    Septic arthritis is encountered very rarely during the neonatal period and its diagnosis can delay because of atypical symptoms, thus it may lead to serious sequelae. The sequale can be prevented by early diagnosis and concomitant treatment. In neonates, pain can be experienced as a result of pseudoparalysis and of movement of the effected joints. A 17-day-old neonatal patient was brought to our hospital with complaint of unrest and then diagnosed with septic arthritis due to propagation of Klebsiella pneumoniae in joint fluid culture was represented because of the rarity of such a case. PMID:27042550

  16. Biochemical and Structural Characterization of a Ureidoglycine Aminotransferase in the Klebsiella pneumoniae Uric Acid Catabolic Pathway

    SciTech Connect

    French, Jarrod B.; Ealick, Steven E.

    2010-09-03

    Many plants, fungi, and bacteria catabolize allantoin as a mechanism for nitrogen assimilation. Recent reports have shown that in plants and some bacteria the product of hydrolysis of allantoin by allantoinase is the unstable intermediate ureidoglycine. While this molecule can spontaneously decay, genetic analysis of some bacterial genomes indicates that an aminotransferase may be present in the pathway. Here we present evidence that Klebsiella pneumoniae HpxJ is an aminotransferase that preferentially converts ureidoglycine and an {alpha}-keto acid into oxalurate and the corresponding amino acid. We determined the crystal structure of HpxJ, allowing us to present an explanation for substrate specificity.

  17. Citrate substitutes for homocitrate in nitrogenase of a nifV mutant of Klebsiella pneumoniae

    SciTech Connect

    Liang, Jihong; Madden, M.; Shah, V.K.; Burris, R.H. )

    1990-09-18

    An organic acid extracted from purified dinitrogenase isolated from a nifV mutant of Klebsiella pneumoniae has been identified as citric acid. H{sub 2} evolution by the citrate-containing dinitrogenase is partially inhibited by CO, and by some substrates for nitrogenase. The response of maximum velocities to changes in pH for both the wild-type and the NifV{sup {minus}} dinitrogenase was compared. No substantial differences between the enzymes were observed, but there are minor differences. Both enzymes are stable in the pH range 4.8-10, but the enzyme activities dropped dramatically below pH 6.2.

  18. Multiclonal expansion of Klebsiella pneumoniae isolates producing-NDM-1 in Rio de Janeiro, Brazil.

    PubMed

    Aires, Caio Augusto Martins; Pereira, Polyana Silva; de Araujo, Carlos Felipe Machado; Chagas, Thiago Pavoni Gomes; Oliveira, Jane Cleide; Buonora, Sibelle Nogueira; Albano, Rodolpho Mattos; Carvalho-Assef, Ana Paula D'Alincourt; Asensi, Marise Dutra

    2017-02-06

    We characterized NDM-1-producing Klebsiella spp. isolates from Rio de Janeiro, Brazil. PCR was applied for resistance and virulence determinants. The genetic context of blaNDM was determined by S1 PFGE and hybridization. The genotyping were performed by PFGE and MLST. Most isolates carried multiple resistance genes and remained susceptible to amikacin, fosfomycin/trometamol, polymyxin B and tigecycline. The spread of NDM-1-producing K. pneumoniae was not associated with a clonal expansion and appears to be associated with Tn3000.

  19. Production and characterization of guluronate lyase from Klebsiella pneumoniae for applications in seaweed biotechnology.

    PubMed

    Ostgaard, K; Knutsen, S H; Dyrset, N; Aasen, I M

    1993-09-01

    Cultures of Klebsiella pneumoniae fermenting sodium alginate produce an extracellular guluronate-specific alginate lyase. This enzyme production was studied in stirred-tank fermentors. Different alginate substrates gave moderate differences in growth and enzyme yield. Alginates with low guluronic content gave reduced biomass but favored enzyme production. Low molecular weight (down to DPn approximately 270) also favored enzyme production. Excessive depolymerization of substrates occurred during heat sterilization of culture media. The enzyme was characterized by its specificity and sensitivity to pH, salt, and calcium. Improved yields of viable protoplasts were documented for Laminaria digitata (Huds.) Lamour.

  20. Simultaneous nosocomial outbreaks caused by multiply resistant Klebsiella pneumoniae types 2 and 30.

    PubMed Central

    Thiemke, W A; Nathan, D M

    1978-01-01

    Two simultaneous Klebsiella pneumoniae outbreaks resistant to multiple antibiotics are reported. The closely related strains were distinguished by comparing the zone diameters obtained by disk diffusion susceptibility testing but not by interpretations of susceptible, resistant, or intermediate. Similar conclusions were made when minimal inhibitory concentrations were considered. The separation was subsequently confirmed by capsular serotyping into types 2 and 30. The data presented suggest that routine antimicrobial susceptibility patterns obtained from disk diffusion and/or minimal inhibitory concentrations may serve as an inexpensive and reliable typing system for epidemiological investigations. PMID:370152

  1. Comparative Genomics of Klebsiella pneumoniae Strains with Different Antibiotic Resistance Profiles▿†

    PubMed Central

    Kumar, Vinod; Sun, Peng; Vamathevan, Jessica; Li, Yong; Ingraham, Karen; Palmer, Leslie; Huang, Jianzhong; Brown, James R.

    2011-01-01

    There is a global emergence of multidrug-resistant (MDR) strains of Klebsiella pneumoniae, a Gram-negative enteric bacterium that causes nosocomial and urinary tract infections. While the epidemiology of K. pneumoniae strains and occurrences of specific antibiotic resistance genes, such as plasmid-borne extended-spectrum β-lactamases (ESBLs), have been extensively studied, only four complete genomes of K. pneumoniae are available. To better understand the multidrug resistance factors in K. pneumoniae, we determined by pyrosequencing the nearly complete genome DNA sequences of two strains with disparate antibiotic resistance profiles, broadly drug-susceptible strain JH1 and strain 1162281, which is resistant to multiple clinically used antibiotics, including extended-spectrum β-lactams, fluoroquinolones, aminoglycosides, trimethoprim, and sulfamethoxazoles. Comparative genomic analysis of JH1, 1162281, and other published K. pneumoniae genomes revealed a core set of 3,631 conserved orthologous proteins, which were used for reconstruction of whole-genome phylogenetic trees. The close evolutionary relationship between JH1 and 1162281 relative to other K. pneumoniae strains suggests that a large component of the genetic and phenotypic diversity of clinical isolates is due to horizontal gene transfer. Using curated lists of over 400 antibiotic resistance genes, we identified all of the elements that differentiated the antibiotic profile of MDR strain 1162281 from that of susceptible strain JH1, such as the presence of additional efflux pumps, ESBLs, and multiple mechanisms of fluoroquinolone resistance. Our study adds new and significant DNA sequence data on K. pneumoniae strains and demonstrates the value of whole-genome sequencing in characterizing multidrug resistance in clinical isolates. PMID:21746949

  2. Biofilm inhibitory effect of chlorhexidine conjugated gold nanoparticles against Klebsiella pneumoniae.

    PubMed

    Ahmed, Ayaz; Khan, Anum Khalid; Anwar, Ayaz; Ali, Syed Abid; Shah, Muhammad Raza

    2016-09-01

    Klebsiella pneumoniae (K. pneumoniae) is one of the major pathogen associated with nosocomial infections, especially catheter associated urinary tract infections which involved biofilm formation. This study was designed to evaluate the antibiofilm efficacy of gold nanoparticle conjugated with chlorhexidine (Au-CHX) against K. pneumoniae isolates. Au-CHX was synthesized and analyzed for stability by using UV-Visible spectrophotometry, atomic force microscopy (AFM), fourier transform infrared spectroscopy (FT-IR) and electrospray ionization mass spectroscopy (ESI-MS). Biofilm inhibition and eradication was performed by crystal violet, 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays and further confirmed by florescence and AFM microscopy. Au-CHX showed the maxima surface plasmon resonance (SPR) band at 535 nm, spherical morphology and polydispersity with size in the range of 20-100 nm. The micro molar concentrations (i.e. 25 and 100 μM) of Au-CHX completely inhibited the biofilm formation and metabolic activity within biofilms of K. pneumoniae reference and three tested clinical isolates, respectively. Time dependant biofilm inhibition assay showed that Au-CHX inhibited the early stage of biofilm formation. While at 75 and 100 μM concentrations, it also eradicated the established biofilms of K. pneumoniae isolates as compared to 2 mM chlorhexidine. Reduced florescence signals and surface roughness during microscopic analysis further confirms the antibiofilm activity of Au-CHX against K. pneumoniae ATCC13882 and clinical isolates. Thus it is concluded that chlorhexidine coated gold nanoparticle not only inhibits the biofilm formation of K. pneumoniae ATCC and clinical isolates but also eradicated the preformed biofilm.

  3. Two unusual cases of successful treatment of hypermucoviscous Klebsiella pneumoniae invasive syndrome.

    PubMed

    Namikawa, Hiroki; Yamada, Koichi; Fujimoto, Hiroki; Oinuma, Ken-Ichi; Tochino, Yoshihiro; Takemoto, Yasuhiko; Kaneko, Yukihiro; Shuto, Taichi; Kakeya, Hiroshi

    2016-11-16

    A few Japanese cases of hypermucoviscous Klebsiella pneumoniae (K. pneumoniae) invasive syndrome have recently been reported. Although extrahepatic complications from bacteremic dissemination have been observed, infected aneurysms are rare. Furthermore, the primary source of infection is generally a liver abscess, and is rarely the prostate. Therefore, we report two atypical cases of hypermucoviscous K. pneumoniae invasive syndrome. The first case was an 81-year-old Japanese man with no significant medical history, who was referred to our hospital for vision loss in his right eye. Contrast-enhanced whole-body computed tomography revealed abscesses in the liver and the prostate, and an infected left internal iliac artery aneurysm. Contrast-enhanced head magnetic resonance imaging revealed brain abscesses. Cultures of the liver abscess specimen and aqueous humor revealed K. pneumoniae with the hypermucoviscosity phenotype, which carried the magA gene (mucoviscosity-associated gene A) and the rmpA gene (regulator of mucoid phenotype A). We performed enucleation of the right eyeball, percutaneous transhepatic drainage, coil embolization of the aneurysm, and administered a 6-week course of antibiotic treatment. The second case was a 69-year-old Japanese man with diabetes mellitus, who was referred to our hospital with fever, pollakiuria, and pain on urination. Contrast-enhanced whole-body computed tomography revealed lung and prostate abscesses, but no liver abscesses. Contrast-enhanced head magnetic resonance imaging revealed brain abscesses. The sputum, urine, prostate abscess specimen, and aqueous humor cultures revealed K. pneumoniae with the hypermucoviscosity phenotype, which carried magA and rmpA. We performed enucleation of the left eyeball, percutaneous drainage of the prostate abscess, and administered a 5-week course of antibiotic treatment. Hypermucoviscous K. pneumoniae can cause infected aneurysms, and the prostate can be the primary site of infection. We

  4. Spectrum of excess mortality due to carbapenem-resistant Klebsiella pneumoniae infections.

    PubMed

    Hauck, C; Cober, E; Richter, S S; Perez, F; Salata, R A; Kalayjian, R C; Watkins, R R; Scalera, N M; Doi, Y; Kaye, K S; Evans, S; Fowler, V G; Bonomo, R A; van Duin, D

    2016-06-01

    Patients infected or colonized with carbapenem-resistant Klebsiella pneumoniae (CRKp) are often chronically and acutely ill, which results in substantial mortality unrelated to infection. Therefore, estimating excess mortality due to CRKp infections is challenging. The Consortium on Resistance against Carbapenems in K. pneumoniae (CRACKLE) is a prospective multicenter study. Here, patients in CRACKLE were evaluated at the time of their first CRKp bloodstream infection (BSI), pneumonia or urinary tract infection (UTI). A control cohort of patients with CRKp urinary colonization without CRKp infection was constructed. Excess hospital mortality was defined as mortality in cases after subtracting mortality in controls. In addition, the adjusted hazard ratios (aHR) for time-to-hospital-mortality at 30 days associated with infection compared with colonization were calculated in Cox proportional hazard models. In the study period, 260 patients with CRKp infections were included in the BSI (90 patients), pneumonia (49 patients) and UTI (121 patients) groups, who were compared with 223 controls. All-cause hospital mortality in controls was 12%. Excess hospital mortality was 27% in both patients with BSI and those with pneumonia. Excess hospital mortality was not observed in patients with UTI. In multivariable analyses, BSI and pneumonia compared with controls were associated with aHR of 2.59 (95% CI 1.52-4.50, p <0.001) and 3.44 (95% CI 1.80-6.48, p <0.001), respectively. In conclusion, in patients with CRKp infection, pneumonia is associated with the highest excess hospital mortality. Patients with BSI have slightly lower excess hospital mortality rates, whereas excess hospital mortality was not observed in hospitalized patients with UTI. Copyright © 2016 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  5. Sources other than unused sawdust can introduce Klebsiella pneumoniae into dairy herds.

    PubMed

    Verbist, B; Piessens, V; Van Nuffel, A; De Vuyst, L; Heyndrickx, M; Herman, L; Van Coillie, E; De Vliegher, S

    2011-06-01

    A longitudinal study was carried out to detect intramammary infections caused by Klebsiella pneumoniae and to identify potential sources of this bacterial species in the environment of the cows. The study was performed in 6 well-managed Belgian dairy herds from May 2008 to May 2009. Monthly (n=13), unused and used sawdust bedding samples as well as individual quarter milk and feces samples were collected from 10 randomly selected cohort cows in each herd. Cases of clinical mastitis of all lactating cows in the 6 herds were also sampled (n=64). From the 3,518 collected samples, 153 K. pneumoniae isolates were obtained, of which 2 originated from milk (clinical mastitis cases). In feces (n=728), used bedding (n=73), and unused bedding (n=73), respectively, 125 (17.2%), 20 (27.4%), and 6 (8.2%) isolates were found. The isolates were fingerprinted by means of pulsed field gel electrophoresis. In total, 109 different pulsotypes were differentiated, indicating a high degree of genetic diversity within the isolates. All isolates from unused bedding belonged to pulsotypes other than those from the other sources, suggesting that sources other than unused sawdust may introduce K. pneumoniae into the herd. Only 2 pulsotypes contained isolates originating from different sources. Pulsotype 10 was found in milk and used bedding and pulsotype 21 was found in feces and used bedding. The 2 milk isolates originated from 2 cows in the same herd but they belonged to a different pulsotype. The results indicate that K. pneumoniae can be prevalent in the environment without causing significant mastitis problems. Most cows were shedding K. pneumoniae in feces, substantiating findings under very different conditions (i.e., American dairy herds). Contamination of used bedding in the cubicles with K. pneumoniae from feces was confirmed, whereas unused bedding was not an important source of K. pneumoniae for the environment of the cows.

  6. TEM and SHV Genes in Klebsiella pneumoniae Isolated from Cockroaches and Their Antimicrobial Resistance Pattern.

    PubMed

    Doosti, Abbas; Pourabbas, Mohammad; Arshi, Asghar; Chehelgerdi, Mohammad; Kabiri, Hamidreza

    2015-02-01

    Klebsiella pneumoniae is a gram-negative rod bacterium, a known cause of community-acquired bacterial pneumonia and is an important hospital-acquired pathogen that causes severe morbidity and mortality. The aim of this study was to identify the TEM and SHV genes in K. pneumoniae isolated from cockroaches obtained from hospitals. In this study, 250 cockroaches were collected from different hospitals in the province of Chaharmahal Va Bakhtiari, which is located in southwest Iran. The samples were examined for the presence of K. pneumoniae by plating onto a combination of culture media, and the antimicrobial susceptibility patterns of isolated K. pneumoniae from samples were evaluated using the disk diffusion test. In addition, from the culture, genomic bacterial DNA was extracted, and sequence-specific targets (TEM and SHV genes) were amplified using the polymerase chain reaction (PCR) method. Out of 250 cockroach samples collected from various hospitals, 179 samples (71.60%) were positive for K. pneumoniae. PCR reaction was performed using specific oligonucleotide primers (TEM-F, TEM-R and SHV-F, SHV-R) for the amplification of each gene, and amplified products were visualized on 1% agarose gel electrophoresis. Of all the specimens amplified by PCR in this research, 32 samples (17.87%) were positive for TEM and 15 samples (8.37%) were positive for SHV. Detection of TEM and SHV genes using molecular methods and their pattern of antimicrobial resistance can provide useful information about the epidemiology of and risk factors associated with K. pneumoniae infection.

  7. Virulent Clones of Klebsiella pneumoniae: Identification and Evolutionary Scenario Based on Genomic and Phenotypic Characterization

    PubMed Central

    Brisse, Sylvain; Fevre, Cindy; Passet, Virginie; Issenhuth-Jeanjean, Sylvie; Tournebize, Régis; Diancourt, Laure; Grimont, Patrick

    2009-01-01

    Klebsiella pneumoniae is found in the environment and as a harmless commensal, but is also a frequent nosocomial pathogen (causing urinary, respiratory and blood infections) and the agent of specific human infections including Friedländer's pneumonia, rhinoscleroma and the emerging disease pyogenic liver abscess (PLA). The identification and precise definition of virulent clones, i.e. groups of strains with a single ancestor that are associated with particular infections, is critical to understand the evolution of pathogenicity from commensalism and for a better control of infections. We analyzed 235 K. pneumoniae isolates of diverse environmental and clinical origins by multilocus sequence typing, virulence gene content, biochemical and capsular profiling and virulence to mice. Phylogenetic analysis of housekeeping genes clearly defined clones that differ sharply by their clinical source and biological features. First, two clones comprising isolates of capsular type K1, clone CC23K1 and clone CC82K1, were strongly associated with PLA and respiratory infection, respectively. Second, only one of the two major disclosed K2 clones was highly virulent to mice. Third, strains associated with the human infections ozena and rhinoscleroma each corresponded to one monomorphic clone. Therefore, K. pneumoniae subsp. ozaenae and K. pneumoniae subsp. rhinoscleromatis should be regarded as virulent clones derived from K. pneumoniae. The lack of strict association of virulent capsular types with clones was explained by horizontal transfer of the cps operon, responsible for the synthesis of the capsular polysaccharide. Finally, the reduction of metabolic versatility observed in clones Rhinoscleromatis, Ozaenae and CC82K1 indicates an evolutionary process of specialization to a pathogenic lifestyle. In contrast, clone CC23K1 remains metabolically versatile, suggesting recent acquisition of invasive potential. In conclusion, our results reveal the existence of important virulent

  8. Host defence during Klebsiella pneumonia relies on haematopoietic-expressed Toll-like receptors 4 and 2.

    PubMed

    Wieland, C W; van Lieshout, M H P; Hoogendijk, A J; van der Poll, T

    2011-04-01

    In this study, the relative roles of Toll-like receptor (TLR)2 and TLR4 were investigated independently and together. Moreover, we studied the role of haematopoietic compartment in anti-Klebsiella host defence. We infected TLR2 and TLR4 single-, and TLR2×4 double knockout (KO) animals with different doses of Klebsiella pneumoniae. In addition, bone marrow chimeric mice were created and infected. TLR4 played a more prominent role in antibacterial defence than TLR2, considering that only TLR4 KO mice demonstrated enhanced bacterial growth in lungs and spleen 24 h after infection with 3×10³ colony-forming units of Klebsiella compared with wild-type (WT) mice. In late-stage infection or after exposure to a higher infectious dose, bacterial counts in lungs of TLR2 KO animals were elevated compared with WT mice and TLR2×4 KO animals were more susceptible to infection than TLR4 KO mice. TLR signalling in cells of haematopoietic origin is of primary importance in host defence against K. pneumoniae. These data suggest that: 1) TLR4 drives the antibacterial host response after induction of pneumonia with relatively low Klebsiella doses; 2) TLR2 becomes involved at a later phase of the infection and/or upon exposure to higher bacterial burdens; and 3) haematopoietic TLR2 and TLR4 are important for an adequate host response during Klebsiella pneumonia.

  9. Dietary supplementation with omega-3 polyunsaturated fatty acids ameliorates acute pneumonia induced by Klebsiella pneumoniae in BALB/c mice.

    PubMed

    Sharma, Sonica; Chhibber, Sanjay; Mohan, Harsh; Sharma, Saroj

    2013-07-01

    The immune benefits associated with the optimal intake of dietary fatty acids are widely known. The objective of the present investigation was to elucidate the role of omega-3 polyunsaturated fatty acids (n-3 PUFA) food source on acute pneumonia induced by Klebsiella pneumoniae. Three different n-3 PUFA preparations (cod liver oil, Maxigard, and flaxseed oil) were orally supplemented and infection was induced in different groups of experimental mice. Mice fed olive oil and normal saline served as oil and saline controls, respectively. After 2 weeks of fatty acid feeding, no effect on the establishment of infection was observed when acute pneumonia was induced in animals. On the other hand, 6 weeks of n-3 PUFA administration was found to improve resistance in mice, as reduced lung bacterial load coupled with significant improvement in pathology was seen in infected mice. Alveolar macrophages collected from all 3 groups of mice fed n-3 PUFA exhibited a significant decrease in the level of apoptosis following infection with K. pneumoniae and an enhanced in vitro phagocytic potential for the pathogen. Lower lung levels of nitric oxide, malondialdehyde, and lactate dehydrogenase were associated with a decrease in the severity of tissue damage. There was a significant increase in the lung levels of pro-inflammatory cytokines (tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β)). No significant change was observed in the levels of interleukin-10 (IL-10). This study highlights that dietary n-3 PUFA supplementation exerts an overall beneficial effect against acute experimental pneumonia. This mechanism is operative through upregulation of nonspecific and specific immune defenses of the host.

  10. Prospectively defined murine mesenchymal stem cells inhibit Klebsiella pneumoniae-induced acute lung injury and improve pneumonia survival.

    PubMed

    Hackstein, Holger; Lippitsch, Anne; Krug, Philipp; Schevtschenko, Inna; Kranz, Sabine; Hecker, Matthias; Dietert, Kristina; Gruber, Achim D; Bein, Gregor; Brendel, Cornelia; Baal, Nelli

    2015-10-06

    Numerous studies have described the immunosuppressive capacity of mesenchymal stem cells (MSC) but these studies use mixtures of heterogeneous progenitor cells for in vitro expansion. Recently, multipotent MSC have been prospectively identified in murine bone marrow (BM) on the basis of PDFGRa(+) SCA1(+) CD45(-) TER119(-) (PαS) expression but the immunomodulatory capacity of these MSC is unknown. We isolated PαS MSC by high-purity FACS sorting of murine BM and after in vitro expansion we analyzed the in vivo immunomodulatory activity during acute pneumonia. PαS MSC (1 × 10(6)) were applied intratracheally 4 h after acute respiratory Klebsiella pneumoniae induced infection. PαS MSC treatment resulted in significantly reduced alveolitis and protein leakage in comparison to mock-treated controls. PαS MSC-treated mice exhibited significantly reduced alveolar TNF-α and IL-12p70 expression, while IL-10 expression was unaffected. Dissection of respiratory dendritic cell (DC) subsets by multiparameter flow cytometry revealed significantly reduced lung DC infiltration and significantly reduced CD86 costimulatory expression on lung CD103(+) DC in PαS MSC-treated mice. In the post-acute phase of pneumonia, PαS MSC-treated animals exhibited significantly reduced respiratory IL-17(+) CD4(+) T cells and IFN-γ(+) CD4(+) T cells. Moreover, PαS MSC treatment significantly improved overall pneumonia survival and did not increase bacterial load. In this study we demonstrated for the first time the feasibility and in vivo immunomodulatory capacity of prospectively defined MSC in pneumonia.

  11. Mechanism of resistance and antibacterial susceptibility in extended-spectrum β-lactamase phenotype Klebsiella pneumoniae and Klebsiella oxytoca isolated between 2000 and 2010 in Japan.

    PubMed

    Sato, Takafumi; Hara, Takafumi; Horiyama, Tsukasa; Kanazawa, Sachi; Yamaguchi, Takahiro; Maki, Hideki

    2015-05-01

    Clinical isolates of Klebsiella pneumoniae and Klebsiella oxytoca collected from 20 Japanese medical facilities between 2000 and 2010 were analysed to evaluate the mechanisms of resistance and antibacterial susceptibilities to 14 antimicrobials. Overall, eight of 484 (1.6%) K. pneumoniae and 19 of 359 (5.3%) K. oxytoca were determined to be extended-spectrum β-lactamase (ESBL) phenotype isolates, and the identified ESBLs amongst the K. pneumoniae isolates were CTX-M-2, -3, -14 and -15, and SHV-12. In contrast, overproduction of chromosomal β-lactamase OXY-2, which was due to a distinct mutation at the - 10 promoter region of this gene, conferred the ESBL phenotype to all the K. oxytoca isolates except one. Based on the Clinical and Laboratory Standards Institute breakpoints, all the ESBL phenotype K. pneumoniae were susceptible to doripenem, flomoxef, moxalactam (latamoxef), cefmetazole and tazobactam/piperacillin, whereas the ESBL phenotype K. oxytoca were susceptible to ceftazidime and ceftibuten in addition to the above, with the exception of tazobactam/piperacillin. Amongst the oral antimicrobials, ceftibuten was relatively effective against both ESBL phenotype Klebsiella species compared with levofloxacin and amoxicillin/clavulanic acid.

  12. Nitrogen fixation by Klebsiella pneumoniae is inhibited by certain multicopy hybrid nif plasmids.

    PubMed Central

    Riedel, G E; Brown, S E; Ausubel, F M

    1983-01-01

    In our studies of nif gene regulation, we have observed that certain hybrid nif plasmids drastically inhibit the expression of the chromosomal nif genes of Klebsiella pneumonia. Wild-type (Nif+) K. pneumoniae strains that acquire certain hybrid nif plasmids also acquire the Nif- phenotype; these strains lose 90 to 99% of all detectable nitrogen fixation activity and grow poorly (or not at all) on solid media with N2 as the sole nitrogen source. We describe experiments which defined this inhibition of the Nif+ phenotype by hybrid nif plasmids and identify and characterize four nif DNA regions associated with this inhibition. We show that plasmids carrying these nif regions could recombine with, but not complement, nif chromosomal mutations. Our results suggest that inhibition of the Nif+ phenotype will provide a useful bioassay for some of the factors that mediate nif gene expression. PMID:6336738

  13. Characterization of OXA-204, a carbapenem-hydrolyzing class D β-lactamase from Klebsiella pneumoniae.

    PubMed

    Potron, Anaïs; Nordmann, Patrice; Poirel, Laurent

    2013-01-01

    A Klebsiella pneumoniae clinical isolate recovered in Tunisia showed resistance to all β-lactams and decreased susceptibility to carbapenems. K. pneumoniae 204 expressed the carbapenem-hydrolyzing β-lactamase OXA-204, differing from OXA-48 by two amino acid substitutions (Gln98His and Thr99Arg) (class D β-lactamase [DBL] numbering). OXA-48 and OXA-204 shared similar resistance profiles, hydrolyzing carbapenems but sparing broad-spectrum cephalosporins. The bla(OXA-204) gene was located on a ca. 150-kb IncA/C-type plasmid, which also carried the bla(CMY-4) gene. The bla(OXA-204) gene was associated with an ISEcp1 element, whereas the bla(OXA-48) genes are usually associated with IS1999.

  14. Open Conversion after Aortic Endograft Infection Caused by Colistin-Resistant, Carbapenemase-Producing Klebsiella pneumoniae.

    PubMed

    Montelione, Nunzio; Menna, Danilo; Sirignano, Pasqualino; Capoccia, Laura; Mansour, Wassim; Speziale, Francesco

    2016-10-01

    A 62-year-old man presented with fever, abdominal pain, and malaise 13 months after emergency endovascular aortic repair. Computed tomographic angiograms showed a periprosthetic fluid and gas collection, so infection was diagnosed. Open conversion was performed, involving endograft explantation and in situ aortic reconstruction. Cultures and the explanted prosthesis were positive for carbapenemase-producing Klebsiella pneumoniae, resistant to colistin. Because of the sparse data on endograft infections caused by this pathogen, we placed the patient on an empiric double-carbapenem regimen for 4 weeks. Symptomatic recovery occurred after 21 days. On the 30th day, we deployed a stent to treat a new pseudoaneurysm. Three years later, the patient had no signs of persistent or recurrent infection. We think that this is the first report of aortic endograft infection caused by colistin-resistant, carbapenemase-producing K. pneumoniae.

  15. Clonal dissemination of Klebsiella pneumoniae ST258 harbouring KPC-2 in Argentina.

    PubMed

    Gomez, S A; Pasteran, F G; Faccone, D; Tijet, N; Rapoport, M; Lucero, C; Lastovetska, O; Albornoz, E; Galas, M; Melano, R G; Corso, A; Petroni, A

    2011-10-01

    The present work describes the abrupt emergence of Klebsiella pneumoniae carbapenemase (KPC) and characterizes the first 79 KPC-producing enterobacteria from Argentina (isolated from 2006 to 2010). The emergence of bla(KPC-2) was characterized by two patterns of dispersion: the first was the sporadic occurrence in diverse enterobacteria from distant geographical regions, harbouring plasmids of different incompatibility groups and bla(KPC-2) in an unusual genetic environment flanked by ISKpn8-Δbla(TEM-1) and ISKpn6-like. bla(KPC-2) was associated with IncL/M transferable plasmids; the second was the abrupt clonal spread of K. pneumoniae ST258 harbouring bla(KPC-2) in Tn4401a.

  16. Role of capsule and O antigen in resistance of Klebsiella pneumoniae to serum bactericidal activity.

    PubMed Central

    Tomás, J M; Benedí, V J; Ciurana, B; Jofre, J

    1986-01-01

    The ability of Klebsiella pneumoniae strains to resist the bactericidal activity of serum was quantitated. The K. pneumoniae strains tested included mutants lacking the capsular polysaccharide and mutants having a modified lipopolysaccharide structure. The last mutants were obtained as phage-resistant mutants, and their lipopolysaccharide was characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and chemical analysis. Serum-resistant mutants derived from phage-resistant mutants (lipopolysaccharide mutants) were also characterized. Resistance to the bactericidal activity of complement was mediated by the lipopolysaccharide, especially by the O-antigen polysaccharide chains. The capsular polysaccharide seemed not to play any important role in resistance to serum bactericidal activity in this bacterium. Images PMID:3531020

  17. Characterization of resistance genes in 68 ESBL-producing Klebsiella pneumonia in Lebanon.

    PubMed

    Charrouf, F Obeid; Hamze, M; Mallat, H; Achkar, M; Dabboussia, F

    2014-12-01

    We studied 68 ESBL-producing Klebsiella pneumoniae strains isolated in Lebanon, determined their profile of resistance to antibiotics, and identified 6 ESBL genes. The susceptibility to antibiotics was determined by the disk diffusion method. The MIC of carbapenems and cefepime was determined by the agar dilution method. ESBL genes were detected by PCR. A percentage of 88.2% and 86.7% of isolates carried the SHV and CTX-M gene respectively; combinations of more than 1 gene of resistance were detected in several isolates. Five strains were resistant to carbapenems; 4/5 carried the OXA-48 gene. Our study revealed the emergence of K. pneumoniae ESBL (+) strains carrying several types of genes involved in this phenotype; we also identified carbapenem-resistant strains due to the OXA-48 gene, which are a real threat for public health, especially in Lebanon.

  18. IMP-Producing Carbapenem-Resistant Klebsiella pneumoniae in the United States ▿

    PubMed Central

    Limbago, Brandi M.; Rasheed, J. Kamile; Anderson, Karen F.; Zhu, Wenming; Kitchel, Brandon; Watz, Nancy; Munro, Susan; Gans, Hayley; Banaei, Niaz; Kallen, Alex J.

    2011-01-01

    The emergence and spread of carbapenem-resistant Enterobacteriaceae (CRE) producing acquired carbapenemases have created a global public health crisis. In the United States, CRE producing the Klebsiella pneumoniae carbapenemase (KPC) are increasingly common and are endemic in some regions. Metallo-β-lactamase (MBL)-producing CRE have recently been reported in the United States among patients who received medical care in countries where such organisms are common. Here, we describe three carbapenem-resistant K. pneumoniae isolates recovered from pediatric patients at a single U.S. health care facility, none of whom had a history of international travel. The isolates were resistant to carbapenems but susceptible to aztreonam, trimethoprim-sulfamethoxazole, and fluoroquinolones. The three isolates were closely related to each other by pulsed-field gel electrophoresis and contained a common plasmid. PCR and sequence analysis confirmed that these isolates produce IMP-4, an MBL carbapenemase not previously published as present among Enterobacteriaceae in the United States. PMID:21998425

  19. Neutral red-mediated microbial electrosynthesis by Escherichia coli, Klebsiella pneumoniae, and Zymomonas mobilis.

    PubMed

    Harrington, Timothy D; Mohamed, Abdelrhman; Tran, Vi N; Biria, Saeid; Gargouri, Mahmoud; Park, Jeong-Jin; Gang, David R; Beyenal, Haluk

    2015-11-01

    The aim of this work was to compare the effects of electrosynthesis on different bacterial species. The effects of neutral red-mediated electrosynthesis on the metabolite profiles of three microorganisms: Escherichia coli, Klebsiella pneumoniae, and Zymomonas mobilis, were measured and compared and contrasted. A statistically comprehensive analysis of neutral red-mediated electrosynthesis is presented using the analysis of end-product profiles, current delivered, and changes in cellular protein expression. K. pneumoniae displayed the most dramatic response to electrosynthesis of the three bacteria, producing 93% more ethanol and 76% more lactate vs. control fermentation with no neutral red and no electron delivery. Z. mobilis showed no response to electrosynthesis except elevated acetate titers. Stoichiometric comparison showed that NAD(+) reduction by neutral red could not account for changes in metabolites during electrosynthesis. Neutral red-mediated electrosynthesis was shown to have multifarious effects on the three species.

  20. Metabolic Engineering of Klebsiella pneumoniae for the Production of 2-Butanone from Glucose

    PubMed Central

    Chen, Zhen; Sun, He; Huang, Jinhai; Wu, Yao; Liu, Dehua

    2015-01-01

    2-Butanone is an important commodity chemical of wide application in different areas. In this study, Klebsiella pneumoniae was engineered to directly produce 2-butanone from glucose by extending its native 2, 3-butanediol synthesis pathway. To identify the potential enzyme for the efficient conversion of 2, 3-butanediol to 2-butanone, we screened different glycerol dehydratases and diol dehydratases. By introducing the diol dehydratase from Lactobacillus brevis and deleting the ldhA gene encoding lactate dehydrogenase, the engineered K. pneumoniae was able to accumulate 246 mg/L of 2-butanone in shake flask. With further optimization of culture condition, the titer of 2-butanone was increased to 450 mg/L. This study lays the basis for developing an efficient biological process for 2-butanone production. PMID:26465746

  1. Neutral red-mediated microbial electrosynthesis by Escherichia coli, Klebsiella pneumoniae, and Zymomonas mobilis

    PubMed Central

    Harrington, Timothy D.; Mohamed, Abdelrhman; Tran, Vi N.; Biria, Saeid; Gargouri, Mahmoud; Park, Jeong-Jin; Gang, David R.; Beyenal, Haluk

    2015-01-01

    The aim of this work was to compare the effects of electrosynthesis on different bacterial species. The effects of neutral red-mediated electrosynthesis on the metabolite profiles of three microorganisms: Escherichia coli, Klebsiella pneumoniae, and Zymomonas mobilis, were measured and compared and contrasted. A statistically comprehensive analysis of neutral red-mediated electrosynthesis is presented using the analysis of end-product profiles, current delivered, and changes in cellular protein expression. K. pneumoniae displayed the most dramatic response to electrosynthesis of the three bacteria, producing 93% more ethanol and 76% more lactate vs. control fermentation with no neutral red and no electron delivery. Z. mobilis showed no response to electrosynthesis except elevated acetate titers. Stoichiometric comparison showed that NAD+ reduction by neutral red could not account for changes in metabolites during electrosynthesis. Neutral red-mediated electrosynthesis was shown to have multifarious effects on the three species. PMID:26096579

  2. Lemierre's Syndrome Caused by Klebsiella pneumoniae in a Diabetic Patient: A Case Report and Review of the Literature

    PubMed Central

    Chuncharunee, Alan

    2015-01-01

    Lemierre's syndrome is characterized by an oropharyngeal infection with internal jugular vein thrombosis followed by metastatic infections in other organs. This infection is usually caused by Fusobacterium spp. In this report, we present a rare case of Klebsiella pneumoniae-associated Lemierre's syndrome in a patient with poorly-controlled diabetes mellitus. The infection was complicated by septic emboli in many organs, which led to the patient's death, despite combined antibiotics, anticoagulant therapy, and surgical intervention. Therein, a literature review was performed for reported cases of Lemierre's syndrome caused by Klebsiella pneumoniae and the results are summarized here. PMID:26279962

  3. Predominance of Klebsiella pneumoniae ST14 carrying CTX-M-15 causing neonatal sepsis in Tanzania.

    PubMed

    Mshana, Stephen E; Hain, Torsten; Domann, Eugen; Lyamuya, Eligius F; Chakraborty, Trinad; Imirzalioglu, Can

    2013-10-07

    Klebsiella pneumoniae strains expressing ESBLs are a predominant cause of hospital acquired infections. Here we describe the molecular epidemiology of these isolates in a tertiary hospital in Tanzania, as potential pathogens for neonatal infections. Between April 2009 and March 2010 all Klebsiella pneumoniae isolates with phenotypic expression Extended Spectrum Beta Lactamase (ESBL) were collected and characterized. Identification was done using in house biochemical tests in case of ambiguous results confirmation was done using API 20E. Susceptibility testing was determined using the disc diffusion method followed by specific PCR and sequencing to determine ESBL genes. Phylogenetic analysis, Pulse field gel electrophoresis (PFGE) and Multi-Locus sequence typing (MLST) to PFGE clusters representative isolates were performed to determine clones of the isolates. Conjugation and hybridization were performed to determine the location of blaCTX-M-15 gene. A total of 92 non-repetitive ESBL producing K. pneumoniae representing 50.3% of Klebsiella pneumoniae isolates were characterized. These isolates were from blood 61 (66%), wound swab 13 (14%), urine 12 (13%) and pus 6 (7%) were analyzed. Most blood culture strains originated from neonatal unit 39/61(64%) and 22 (36%) of the blood culture isolates were from neonatal ICU. All isolates were resistant to gentamicin and 54% were resistant to ciprofloxacin. Using a similarity index of 80%, the isolates were assigned to thirteen clusters based on PFGE patterns and contained sub-clusters with identical strains indicating clonal outbreaks. Cluster X5, X7 and X8, and X9 were grouped into ST48, ST14 and ST348 respectively. Based on gyrA PCR- RFLP phylogenetic analysis all isolates were grouped as KpI. The predominant ESBL allele detected was blaCTX-M-15 which was found in 76% of isolates, followed by blaTEM-104 (19%), blaSHV-11 (3.2%) and blaTEM-176 (2%). The blaCTX-M-15 gene was located in multiple conjugative IncF plasmids

  4. Molecular Characterization of Klebsiella pneumoniae Carbapenemase (KPC)-Producing Enterobacteriaceae in Ontario, Canada, 2008-2011

    PubMed Central

    Tijet, Nathalie; Sheth, Prameet M.; Lastovetska, Olga; Chung, Catherine; Patel, Samir N.; Melano, Roberto G.

    2014-01-01

    Due to the lack of detailed reports of Klebsiella pneumoniae carbapenemase (KPC)-producing enterobacteria in Ontario, Canada, we perform a molecular characterization of KPC-producing Enterobacteriaceae submitted to the provincial reference laboratory from 2008 to 2011. Susceptibility profiles were accessed by E-test. Molecular types of isolates were determined by pulse-field gel electrophoresis (PFGE) and multilocus sequence typing. Screening of ß-lactamase genes was performed by multiplex PCR and alleles were identified by DNA sequencing. The genetic platform of blaKPC gene was analyzed by PCR. Plasmid replicons were typed using PCR-based typing approach. KPC-plasmids were also evaluated by S1 nuclease-PFGE and Southern blot. Thirty unique clinical isolates (26 Klebsiella pneumoniae, 2 Enterobacter cloacae, 1 Citrobacter freundii and 1 Raoultella ornithinolytica) were identified as blaKPC positive: 4 in 2008, 3 in 2009, 10 in 2010 and 13 in 2011. The majority exhibited resistance to carbapenems, cephalosporins and fluoroquinolones and two isolates were also resistant to colistin. The isolates harbored blaKPC-2 (n = 23) or blaKPC-3 (n = 7). blaTEM-1 (n = 27) was commonly detected and occasionally blaOXA-1 (n = 3) and blaCTX-M-15 (n = 1). As expected, all K. pneumoniae isolates carried blaSHV-11. blaKPC genes were identified on Tn4401a (n = 20) or b (n = 10) isoforms, on plasmids of different sizes belonging to the incompatibility groups IncFIIA (n = 19), IncN (n = 3), IncI2 (n = 3), IncFrep (n = 2) and IncA/C (n = 1). The occurrence of KPC ß-lactamase in Ontario was mainly associated with the spread of the K. pneumoniae clone ST258. PMID:25549365

  5. A liver abscess deprived a healthy adult of eyesight: endogenous endophthalmitis associated with a pyogenic liver abscess caused by serotype K1 Klebsiella pneumonia.

    PubMed

    Maruno, Takahisa; Ooiwa, Yoko; Takahashi, Ken; Kodama, Yuzo; Takakura, Shunji; Ichiyama, Satoshi; Chiba, Tsutomu

    2013-01-01

    Klebsiella pneumonia usually causes urinary tract infections, pneumonia, and other infectious diseases in hospitalized and immunocompromised patients. Among the types of Klebsiella pneumonia, serotype K1 is known to be a highly virulent pathogen. We herein report the case of a healthy 63-year-old man with a pyogenic liver abscess and bilateral endogenous endophthalmitis caused by serotype K1 Klebsiella pneumonia. Although the patient received percutaneous abscess drainage and antibiotic therapy, he lost his eyesight. To improve the poor prognoses of ocular complications, providing both an earlier diagnosis and treatment is critical.

  6. Frequency of Klebsiella pneumoniae carbapenemase (KPC) and non-KPC-producing Klebsiella contamination of Healthcare workers and the environment

    PubMed Central

    Rock, Clare; Thom, Kerri A.; Masnick, Max; Johnson, J. Kristie; Harris, Anthony D.; Morgan, Daniel J

    2014-01-01

    We examined contamination of healthcare worker (HCW) gown and gloves after caring for patients with Klebsiella Producing Carbapenemase-producing and non-KPC-producing Klebsiella as a proxy for horizontal transmission. Contamination rate with Klebsiella is similar to MRSA and VRE, with 14% (31/220) of HCW-patient interactions resulting in contamination of gloves and gowns. PMID:24602950

  7. [Protective effect of Lactobacillus acidophilus on development of infection, caused by Klebsiella pneumoniae].

    PubMed

    Kostiuk, O P; Chernyshova, L I; Slukvin, I I

    1993-01-01

    The mechanisms of protective action of Lactobacillus have been studied during development of the generalized infection induced by Klebsiella pneumoniae in CBA mice after weaning. The mice were infected intragastrically during the first day after weaning (1 x 10(9) bacterias per mice). Suspensions of Lactobacillus were introduced before and after infection during 10 days (1 x 10(6) bacterias per mice). It has been shown that introduction of Lactobacillus substantially decreased the level of the gut contamination by Klebsiella, prevented generalization of infection and death of animals. Significant higher levels of IgA in the blood serum, IgA and IgM in the gut content, percentage of splenocytes, expressing surface IgM and IgG were observed on the 7th day as compared with those in animals without Lactobacillus. Significantly lower percentage of splenocytes, expressing CD4 antigen was also observed. On the 11th day after infection the mice receiving lactobacillus have shown a tendency to an increase of IgA in the gut content, significantly lower concentrations of IgM in the gut content and a higher level of IgA to the blood serum as compared with the control. Other characteristics were comparable to those of the control group. A conclusion is made that introduction of Lactobacillus prevents development of the Klebsiella infection and protects the immune system from excessive antigenic action.

  8. Short Chain N-Acylhomoserine Lactone Production by Clinical Multidrug Resistant Klebsiella pneumoniae Strain CSG20

    PubMed Central

    Ngeow, Yun Fong; Cheng, Huey Jia; Chen, Jian Woon; Yin, Wai-Fong; Chan, Kok-Gan

    2013-01-01

    Klebsiella pneumoniae is one of the most common Gram-negative bacterial pathogens in clinical practice. It is associated with a wide range of disorders, ranging from superficial skin and soft tissue infections to potentially fatal sepsis in the lungs and blood stream. Quorum sensing, or bacterial cell-cell communication, refers to population density-dependent gene expression modulation. Quorum sensing in Proteobacteria relies on the production and sensing of signaling molecules which are mostly N-acylhomoserine lactones. Here, we report the identification of a multidrug resistant clinical isolate, K. pneumoniae strain CSG20, using matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry. We further confirmed quorum sensing activity in this strain with the use of high resolution tandem liquid chromatography quadrupole mass spectrometry and provided evidence K. pneumoniae strain CSG20 produced N-hexanoyl-homoserine lactone (C6-HSL). To the best of our knowledge, this is the first report on the production of N-hexanoylhomoserine lactone (C6-HSL) in clinical isolate K. pneumoniae. PMID:24284772

  9. Production of Chemicals by Klebsiella pneumoniae Using Bamboo Hydrolysate as Feedstock.

    PubMed

    Wei, Dong; Gu, Jinjie; Zhang, Zhongxi; Wang, Chenhong; Wang, Dexin; Kim, Chul Ho; Jiang, Biao; Shi, Jiping; Hao, Jian

    2017-06-29

    Bamboo is an important biomass, and bamboo hydrolysate is used by Klebsiella pneumoniae as a feedstock for chemical production. Here, bamboo powder was pretreated with NaOH and washed to a neutral pH. Cellulase was added to the pretreated bamboo powder to generate the hydrolysate, which contained 30 g/L glucose and 15 g/L xylose and was used as the carbon source to prepare a medium for chemical production. When cultured in microaerobic conditions, 12.7 g/L 2,3-butanediol was produced by wildtype K. pneumoniae. In aerobic conditions, 13.0 g/L R-acetoin was produced by the budC mutant of K. pneumoniae. A mixture of 25.5 g/L 2-ketogluconic acid and 13.6 g/L xylonic acid was produced by the budA mutant of K. pneumoniae in a two-stage, pH-controlled fermentation with high air supplementation. In the first stage of fermentation, the culture was maintained at a neutral pH; after cell growth, the fermentation proceeded to the second stage, during which the culture was allowed to become acidic.

  10. Short chain N-acylhomoserine lactone production by clinical multidrug resistant Klebsiella pneumoniae strain CSG20.

    PubMed

    Ngeow, Yun Fong; Cheng, Huey Jia; Chen, Jian Woon; Yin, Wai-Fong; Chan, Kok-Gan

    2013-11-07

    Klebsiella pneumoniae is one of the most common Gram-negative bacterial pathogens in clinical practice. It is associated with a wide range of disorders, ranging from superficial skin and soft tissue infections to potentially fatal sepsis in the lungs and blood stream. Quorum sensing, or bacterial cell-cell communication, refers to population density-dependent gene expression modulation. Quorum sensing in Proteobacteria relies on the production and sensing of signaling molecules which are mostly N-acylhomoserine lactones. Here, we report the identification of a multidrug resistant clinical isolate, K. pneumoniae strain CSG20, using matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry. We further confirmed quorum sensing activity in this strain with the use of high resolution tandem liquid chromatography quadrupole mass spectrometry and provided evidence K. pneumoniae strain CSG20 produced N-hexanoyl-homoserine lactone (C6-HSL). To the best of our knowledge, this is the first report on the production of N-hexanoylhomoserine lactone (C6-HSL) in clinical isolate K. pneumoniae.

  11. First clinical cases of NDM-1-producing Klebsiella pneumoniae from two hospitals in Bulgaria.

    PubMed

    Todorova, Bozhana; Sabtcheva, Stefana; Ivanov, Ivan N; Lesseva, Magdalena; Chalashkanov, Tzano; Ioneva, Magda; Bachvarova, Assya; Dobreva, Elina; Kantardjiev, Todor

    2016-12-01

    We report the first confirmed cases of NDM-1-producing Klebsiella pneumoniae infections in two hospitals in Bulgaria. The isolates were diverse in terms of plasmid and co-resistance gene content. K. pneumoniae PR2682, causing sepsis in patient with polytrauma due to traffic accident, harbored blaNDM-1,blaCMY-4, blaCTX-M-15, blaSHV-1, blaTEM-1b, qnrB, and aac(6')-Ib. blaNDM-1 was transferable by conjugation and located on an IncA/C plasmid of 176-kb, which also carried blaCMY-4, blaCTX-M-15, blaTEM-1b, and qnrB. K. pneumoniae PR2830, causing urinary tract infection in prostate cancer patient, harbored blaNDM-1,blaSHV-1, blaTEM-1, and aac(6')-Ib. blaNDM-1 was carried on an 86-kb IncA/C plasmid transferable by conjugation together with blaTEM-1, and aac(6')-Ib. Multilocus sequence typing indicated that the two isolates belonged to sequence type ST11. The emergence of NDM-1-producing K. pneumoniae indicates that blaNDM-1-mediated resistance is already disseminated among Enterobacteriaceae in Bulgaria. Our results further confirm the role of the Balkans as a secondary reservoir where NDM-encoding genes originate.

  12. Inhibition of Klebsiella pneumoniae DnaB helicase by the flavonol galangin.

    PubMed

    Chen, Cheng-Chieh; Huang, Cheng-Yang

    2011-01-01

    Klebsiella pneumoniae is a ubiquitous opportunistic pathogen that colonizes at the mucosal surfaces in humans and causes severe diseases. Many clinical strains of K. pneumoniae are highly resistant to antibiotics. Here, we used fluorescence quenching to show that the flavonols galangin, myricetin, quercetin, and kaempferol, bearing different numbers of hydroxyl substituent on the aromatic rings, may inhibit dNTP binding of the primary replicative DnaB helicase of K. pneumoniae (KpDnaB), an essential component of the cellular replication machinery critical for bacterial survival. The binding affinity of KpDnaB to dNTPs varies in the following order: dCTP ~ dGTP > dTTP > dATP. Addition of 10 μM galangin significantly decreased the binding ability of KpDnaB to dATP, whereas the binding affinity of KpDnaB to dGTP that was almost unaffected. Our analyses suggest that these flavonol compounds may be used in the development of new antibiotics that target K. pneumoniae and other bacteria.

  13. Extended-Spectrum Cephalosporin Compared to Cefazolin for Treatment of Klebsiella pneumoniae-Caused Liver Abscess

    PubMed Central

    Cheng, Hsiao-Pei; Siu, L. K.; Chang, Feng-Yee

    2003-01-01

    From January 1995 to May 2000, a total of 107 adults with liver abscess due to Klebsiella pneumoniae admitted at a large medical center in northern Taiwan were reviewed. Patients were considered to have received cefazolin or an extended-spectrum cephalosporin if they received at least 3 days of that antibiotic within the first 5 days of hospitalization. Fifty-nine (55.1%) patients received cefazolin, and 48 (44.9%) patients received an extended-spectrum cephalosporin. The demographic data, clinical features, severities of illness, and rates of early drainage for the two groups were comparable. However, the rates of developing complications for the two groups were significantly different (37.3 versus 6.3%, respectively; P < 0.001). Furthermore, six independent factors preventing severe complications following liver abscess due to K. pneumoniae were identified: normal platelet count, alkaline phosphatase less than 300 U/liter, no gas formation in the abscess, APACHE III score less than 40, use of an extended-spectrum cephalosporin, and early drainage. In conclusion, cefazolin therapy may be suboptimal for patients with liver abscess due to K. pneumoniae despite active in vitro susceptibility. Use of an extended-spectrum cephalosporin and early drainage for patients with liver abscess due to K. pneumoniae are suggested. PMID:12821451

  14. Persistence of Klebsiella pneumoniae on simulated biofilm in a model drinking water system.

    PubMed

    Szabo, Jeffrey G; Rice, Eugene W; Bishop, Paul L

    2006-08-15

    Persistence of Klebsiella pneumoniae on corroded iron surfaces in drinking water was studied using biofilm annular reactors operated under oligotrophic conditions. Reactors were inoculated with K. pneumoniae, and persistence was monitored in the bulk and biofilm phases. Initial cell concentration of 10(6) MPN/mL in the bulkwater phase resulted in significantly longer adhesion than initial concentrations 1 and 2 orders of magnitude lower. K. pneumoniae cultured in low nutrient growth medium persisted longer in dechlorinated tap water than those cultured in full strength medium. Cell surface charge was more negative under low nutrient conditions, and this influenced electrostatic attraction between the cells and the oxidized iron surface. Cells grown in full strength media persisted longer in water with both low (<0.2 mg/L) and high (>0.5 mg/L) free chlorine residuals. Growth media injected with the cells dechlorinated the water allowing adhesion without inactivation. Microelectrode measurements showed a 40-70% drop in free chlorine from the bulk to the coupon surface, which decreased disinfectant potency against adhered cells. Growth and injection conditions clearly influenced cell adhesion and persistence, but permanent colonization of the corroded iron surface by K. pneumoniae was not observed.

  15. Inhaled fluticasone propionate impairs pulmonary clearance of Klebsiella Pneumoniae in mice

    PubMed Central

    2012-01-01

    Background Recent trials demonstrate increased pneumonia risk in chronic obstructive pulmonary disease patients treated with the inhaled corticosteroid (ICS) fluticasone propionate (FP). There is limited work describing FP effects on host defenses against bacterial pneumonia. Methods C57BL/6 mice received daily, nose-only exposure to nebulized FP or vehicle for 8 days, followed by pulmonary challenge with Klebsiella pneumoniae. Bacterial burden, phagocytosis, leukocyte recruitment, cytokine expression, nitric oxide release, and survival were measured. Results Inhaled FP increased bacterial burden in lungs and blood 48 h after infection but affected neither in vivo phagocytosis of bacteria by alveolar macrophages (AM) nor alveolar neutrophil recruitment. AM from FP-treated mice showed impaired expression of infection induced TNF-alpha, IP-10 (CXCL-10), and interleukin 6 (IL-6), and AM also showed a trend towards impaired intracellular pathogen control following in vivo infection. In vitro FP treatment resulted in a dose-dependent impairment of cytokine expression by AM. Furthermore, infection-induced nitric oxide (but not hydrogen peroxide) production was impaired by FP in vivo and in vitro. FP decreased survival in this model. Conclusions Exposure to inhaled FP impairs pulmonary clearance of K. pneumoniae in mice, an effect associated with greater systemic bacteremia and death. Decreased AM cytokine and nitric oxide expression parallel the failure to control infection. These results support the study of ICS effects on human pulmonary host defenses. PMID:22651370

  16. Klebsiella pneumoniae necrotizing fasciitis of the leg in an elderly French woman.

    PubMed

    Monié, Marguerite; Drieux, Laurence; Nzili, Bernadette; Dicko, Michèle; Goursot, Catherine; Greffard, Sandrine; Decré, Dominique; Mézière, Anthony

    2014-01-01

    Klebsiella pneumoniae necrotizing fasciitis is a rare infection in regions outside of Asia. Here, we present a case of necrotizing fasciitis of the leg caused by K. pneumoniae in a 92-year-old French woman hospitalized in a geriatric rehabilitation unit. The patient initially presented with dermohypodermitis of the leg that developed from a dirty wound following a fall. A few hours later, this painful injury extended to the entire lower limb, with purplish discoloration of the skin, bullae, and necrosis. Septic shock rapidly appeared and the patient died 9 hours after the onset of symptoms. The patient was Caucasian, with no history of travel to Asia or any underlying disease. Computed tomography revealed no infectious metastatic loci. Blood cultures showed growth of capsular serotype K2 K. pneumoniae strains with virulence factors RmpA, yersiniabactin and aerobactin. This rare and fatal case of necrotizing fasciitis caused by a virulent strain of K. pneumoniae occurred in a hospitalized elderly woman without risk factors. Clinicians and geriatricians in particular should be aware of this important albeit unusual differential diagnosis.

  17. Carbapenemase-Producing Klebsiella pneumoniae, a Key Pathogen Set for Global Nosocomial Dominance

    PubMed Central

    Nordmann, Patrice; Poirel, Laurent

    2015-01-01

    The management of infections due to Klebsiella pneumoniae has been complicated by the emergence of antimicrobial resistance, especially to carbapenems. Resistance to carbapenems in K. pneumoniae involves multiple mechanisms, including the production of carbapenemases (e.g., KPC, NDM, VIM, OXA-48-like), as well as alterations in outer membrane permeability mediated by the loss of porins and the upregulation of efflux systems. The latter two mechanisms are often combined with high levels of other types of β-lactamases (e.g., AmpC). K. pneumoniae sequence type 258 (ST258) emerged during the early to mid-2000s as an important human pathogen and has spread extensively throughout the world. ST258 comprises two distinct lineages, namely, clades I and II, and it seems that ST258 is a hybrid clone that was created by a large recombination event between ST11 and ST442. Incompatibility group F plasmids with blaKPC have contributed significantly to the success of ST258. The optimal treatment of infections due to carbapenemase-producing K. pneumoniae remains unknown. Some newer agents show promise for treating infections due to KPC producers; however, effective options for the treatment of NDM producers remain elusive. PMID:26169401

  18. [Carbapenemases among Klebsiella pneumoniae: sensitivity, E-test and Hodge test].

    PubMed

    Essayagh, Touria; Karimou, Aichatou; Elhamzaoui, Sakina

    2012-01-01

    Carbapenems are the major weapons in the treatment of nosocomial infections. Since 1996, there have been an emergence and dissemination of carbapenem resistance in the world, especially among Enterobacteriaceae. Resistance is due to one or several mechanisms including production of carbapenemases. A prospective study was carried out from April to September 2011 at the Laboratory of Microbiology of Military Teaching Hospital in Rabat. The aim of this study was to search by phenotypic methods carbapenemases production in Klebsiella pneumoniae at Laboratory of Microbiology of the HMIMV of Rabat by disc diffusion for susceptibility to imipenem (IMP) and ertapenem (ETP), E-test and Hodge test; 211 strains of K. pneumoniae were analyzed. The samples were most frequently obtained from the urinary tract (63.8%); pus (16.5%) and blood cultures (11.8%); 27% of the strains produced extended spectrum of beta-lactamase (ESBL); 3 strains were resistant to ertapenem by disc diffusion. Investigations of these 3 strains in E-test shown that they were resistant to ertapenem but susceptible to imipenem. Tested by Hodge method, 3 strains gave repeatedly positive results. Thus, rate of K. pneumoniae producing KPC type in our study is 1.42%. Our study shows the evidence of K. pneumoniae type carbapenemase (KPC) in Military Teaching Hospital of Rabat. However, the gene encoding the type of resistance must be determined by molecular methods.

  19. Expression of the Klebsiella pneumoniae CG21 acetoin reductase gene in Clostridium acetobutylicum ATCC 824.

    PubMed

    Wardwell, S A; Yang, Y T; Chang, H Y; San, K Y; Rudolph, F B; Bennett, G N

    2001-10-01

    Acetoin reductase catalyzes the production of 2,3-butanediol from acetoin. The gene encoding the acetoin reductase of Klebsiella pneumoniae CG21 was cloned and expressed in Escherichia coli and Clostridium acetobutylicum ATCC 824. The nucleotide sequence of the gene encoding the enzyme was determined to be 768 bp long. Expression of the K. pneumoniae acetoin reductase gene in E. coli revealed that the enzyme has a molecular mass of about 31,000 Da based on sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. The K. pneumoniae acetoin reductase gene was cloned into a clostridial/E. coli shuttle vector, and expression of the gene resulted in detectable levels of acetoin reductase activity in both E. coli and C. acetobutylicum. While acetoin, the natural substrate of acetoin reductase, is a typical product of fermentation by C. acetobutylicum, 2,3-butanediol is not. Analysis of culture supernatants by gas chromatography revealed that introduction of the K. pneumoniae acetoin reductase gene into C. acetobutylicum was not sufficient for 2,3-butanediol production even though the cultures were producing acetoin. 2,3-Butanediol was produced by cultures of C. acetobutylicum containing the gene only when commercial acetoin was added.

  20. High Production of 3-Hydroxypropionic Acid in Klebsiella pneumoniae by Systematic Optimization of Glycerol Metabolism

    PubMed Central

    Li, Ying; Wang, Xi; Ge, Xizhen; Tian, Pingfang

    2016-01-01

    3-Hydroxypropionic acid (3-HP) is an important platform chemical proposed by the United States Department of Energy. 3-HP can be converted to a series of bulk chemicals. Biological production of 3-HP has made great progress in recent years. However, low yield of 3-HP restricts its commercialization. In this study, systematic optimization was conducted towards high-yield production of 3-HP in Klebsiella pneumoniae. We first investigated appropriate promoters for the key enzyme (aldehyde dehydrogenase, ALDH) in 3-HP biosynthesis, and found that IPTG-inducible tac promoter enabled overexpression of an endogenous ALDH (PuuC) in K. pneumoniae. We optimized the metabolic flux and found that blocking the synthesis of lactic acid and acetic acid significantly increased the production of 3-HP. Additionally, fermentation conditions were optimized and scaled-up cultivation were investigated. The highest 3-HP titer was observed at 83.8 g/L with a high conversion ratio of 54% on substrate glycerol. Furthermore, a flux distribution model of glycerol metabolism in K. pneumoniae was proposed based on in silico analysis. To our knowledge, this is the highest 3-HP production in K. pneumoniae. This work has significantly advanced biological production of 3-HP from renewable carbon sources. PMID:27230116

  1. A Thermostabilized, One-Step PCR Assay for Simultaneous Detection of Klebsiella pneumoniae and Haemophilus influenzae

    PubMed Central

    Khazani, Nur Amalina; Noor, Nik Zuraina Nik Mohd; Yean Yean, Chan

    2017-01-01

    Klebsiella pneumoniae and Haemophilus influenzae are two common pathogens associated with respiratory tract infections. The identification of these pathogens using conventional molecular diagnostic tests requires trained personnel, cold-chain transportation, and storage-dependance, which does not render them user-friendly. The aim of this study was to develop a thermostabilized, cold-chain-free, one-step multiplex PCR for simultaneous detection of K. pneumoniae and H. influenzae. The multiplex PCR assay was designed to amplify the php gene of K. pneumoniae (202 bp) and p6 gene of H. influenzae (582 bp). In addition, the specific primer to amplify glm gene of Helicobacter pylori (105 bp) was included as an internal amplification control. Subsequently, the designed primers and all PCR reagents were thermostabilized by lyophilization. The stability of the thermostabilized PCR was evaluated using the Q10 method. The sensitivity and specificity of performances for thermostabilized PCR were evaluated using 127 clinical isolates and were found to be 100% sensitive and specific. The thermostabilized PCR mix was found to be stable for 30 days and the Q10 accelerated stability was found to be 3.02 months. A cold-chain-free, PCR assay for easy, rapid, and simultaneous detection of K. pneumoniae and H. influenzae was successfully developed in this study. PMID:28386286

  2. Quinolone therapy of Klebsiella pneumoniae sepsis following irradiation: Comparison of pefloxacin, ciprofloxacin, and ofloxacin

    SciTech Connect

    Brook, I.; Elliott, T.B.; Ledney, G.D. )

    1990-05-01

    Exposure to whole-body irradiation is associated with fatal gram-negative sepsis. The effect of oral therapy with three quinolones, pefloxacin, ciprofloxacin, and ofloxacin, for orally acquired Klebsiella pneumoniae infection was tested in B6D2F1 mice exposed to 8.0 Gy whole-body irradiation from bilaterally positioned 60Co sources. A dose of 10(8) organisms was given orally 2 days after irradiation, and therapy was started 1 day later. Quinolones reduced colonization of the ileum with K. pneumoniae: 16 of 28 (57%) untreated mice harbored the organisms, compared to only 12 of 90 (13%) mice treated with quinolones (P less than 0.005). K. pneumoniae was isolated from the livers of 6 of 28 untreated mice, compared to only 1 of 90 treated mice (P less than 0.001). Only 5 of 20 (25%) untreated mice survived for at least 30 days compared with 17 of 20 (85%) mice treated with ofloxacin, 15 of 20 (75%) mice treated with pefloxacin, and 14 of 20 (70%) treated with ciprofloxacin (P less than 0.05). These data illustrate the efficacy of quinolones for oral therapy of orally acquired K. pneumoniae infection in irradiated hosts.

  3. Clonal dissemination of multilocus sequence type ST15 KPC-2-producing Klebsiella pneumoniae in Bulgaria.

    PubMed

    Markovska, Rumyana; Stoeva, Temenuga; Schneider, Ines; Boyanova, Lyudmila; Popova, Valentina; Dacheva, Daniela; Kaneva, Radka; Bauernfeind, Adolf; Mitev, Vanyo; Mitov, Ivan

    2015-10-01

    A total of 36 consecutive clinical and two fecal-screening carbapenem-resistant Klebsiella pneumoniae isolates from two Bulgarian university hospitals (Varna and Pleven) were investigated. Susceptibility testing, conjugation experiments, and plasmid replicon typing were carried out. Beta-lactamases were characterized by isoelectric focusing, PCR, and sequencing. Clonal relatedness was investigated by RAPD and multilocus sequence typing (MLST). Most of the isolates demonstrated multidrug resistance profile. Amikacin and tigecycline retained good activity with susceptibility rates of 95 and 87%, respectively. The resistance rate to colistin was 63%. Six RAPD- and MLST-types were identified: the dominating MLST-type was ST15 (27 isolates), followed by ST76 (six isolates), and ST1350 (two isolates). ST101, ST258, and ST151 were detected once. All except one of the K. pneumoniae produced KPC-2, mostly in combination with CTX-M-15, while for one isolate (ST101) the enzymes OXA-48 and CTX-M-14 were found. All KPC-2-producing transconjugants revealed the presence of IncFII plasmid. The OXA-48- and CTX-M-14-producing isolate showed the presence of L/M replicon type. The dissemination of KPC-2-producing K.pneumoniae in Bulgaria is mainly due to the sustained spread of successful ST15 clone and to a lesser extent of ST76 clone. This is the first report of OXA-48 producing ST101 K. pneumoniae in Bulgaria.

  4. Virulence factors and clinical patterns of hypermucoviscous Klebsiella pneumoniae isolated from urine.

    PubMed

    Kim, Youn Jeong; Kim, Sang Il; Kim, Yang Ree; Wie, Seong Heon; Lee, Hae Kyung; Kim, Soo-Young; Park, Yeon-Joon

    2017-03-01

    Klebsiella pneumoniae with hypermucoviscosity (HM) phenotype is generally more virulent than HM-negative strains. The aim of this study was to investigate the prevalence of HM phenotype among urinary isolates and to compare the virulence factors, antimicrobial susceptibility patterns and clinical characteristics of HM-positive and -negative K. pneumoniae isolated from urine of hospitalized patients. From June to October 2013, a total of 81 non-repetitive K. pneumoniae strains were isolated from urine. HM phenotype was determined by a string test. The K1 and K2 genotypes, the allS, kfu, rmpA, rmpA2 and wabG, aerobactin gene were detected by polymerase chain reaction. Of the 81 K. pneumoniae isolates, 12.3% produced a positive string test. The aerobactin (80.0%[8/10] vs. 15.5%[11/71], p = .0001), allS (40.0%[4/10] vs. 9.9%[7/71], p = .009), rmpA (70.0%[7/10] vs. 14.1%[10/71], p = .0001) and rmpA2 (60.0%[6/10] vs. 16.9%[12/71], p = .002) genes were more prevalent in HM positive than in HM negative strains. The K1 (20.0%[2/10) vs. 8.5%[6/71

  5. Increased susceptibility to Klebsiella pneumonia and mortality in GSNOR-deficient mice.

    PubMed

    Tang, Chi-Hui; Seeley, Eric J; Huang, Xiaozhu; Wolters, Paul J; Liu, Limin

    2013-12-06

    S-nitrosoglutathione reductase (GSNOR) is a key denitrosylase and critically important for protecting immune and other cells from nitrosative stress. Pharmacological inhibition of GSNOR is being actively pursued as a therapeutic approach to increase S-nitrosoglutathione levels for the treatment of asthma and cystic fibrosis. In the present study, we employed GSNOR-deficient (GSNOR(-/-)) mice to investigate whether inactivation of GSNOR may increase susceptibility to pulmonary infection by Klebsiella pneumoniae, a common cause of nosocomial pneumonia. We found that compared to wild-type mice, bacterial colony forming units 48 h after intranasal infection with K. pneumoniae were increased over 4-folds in lung and spleen and strikingly, over a 1000-folds in blood of GSNOR(-/-) mice. Lung injury was comparable between infected wild-type and GSNOR(-/-) mice, but inflammation and injury was significantly elevated in spleen of GSNOR(-/-) mice. Whereas all wild-type mice survived 48 h after infection, 10 of 23 GSNOR(-/-) mice died. Thus, GSNOR appears to play a crucial role in controlling pulmonary and systemic infection by K. pneumoniae. Our results suggest that patients treated in clinical trials with inhibitors of GSNOR should be carefully monitored for signs of infection. Copyright © 2013 Elsevier Inc. All rights reserved.

  6. NDM-1- and OXA-163-producing Klebsiella pneumoniae isolates in Cairo, Egypt, 2012.

    PubMed

    Abdelaziz, Mohammed O; Bonura, Celestino; Aleo, Aurora; Fasciana, Teresa; Mammina, Caterina

    2013-12-01

    Here we describe carbapenem resistance determinants in two Klebsiella pneumoniae isolates recovered from two hospitalised patients in the same intensive care unit of a cancer hospital in Cairo, Egypt. PCR and sequencing were used to detect and characterise β-lactamase genes. Clonal relationships between the isolates were analysed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The first K. pneumoniae isolate carried the blaNDM-1 gene and the second isolate carried the blaOXA-163 gene. Both isolates co-expressed the extended-spectrum β-lactamase CTX-M-15. The two isolates belonged to different sequence types (STs), ST11 and ST16, respectively. No history of travel was established for the two patients. The first identification of NDM-1-producing K. pneumoniae in Egypt adds further evidence to the spread of NDM-1-producing Gram-negative micro-organisms in North Africa. The additional detection of blaOXA-163 in a K. pneumoniae isolate confirms its endemic presence in a critical healthcare setting of this geographic area.

  7. Ampicillin and amoxicillin use and the risk of Klebsiella pneumoniae liver abscess in Taiwan.

    PubMed

    Lin, Yi-Tsung; Liu, Chia-Jen; Yeh, Yi-Chen; Chen, Tzeng-Ji; Fung, Chang-Phone

    2013-07-15

    Klebsiella pneumoniae liver abscess (KPLA) is prevalent in East Asia. Use of amoxicillin/ampicillin may lead to overgrowth of K. pneumoniae in the intestine and predispose to KPLA. We used an animal study and nationwide population-based database to investigate the association between ampicillin/amoxicillin use and KPLA in Taiwan. In an animal study, ampicillin or sterile water was administered orogastrically in serotype K1 K. pneumoniae-colonized mice and the outcome was compared. We identified 855 cases with liver abscess and selected 3420 age- and sex-matched control subjects from the National Health Insurance Research Database. Conditional logistic regression was used to estimate the adjusted odds ratios (ORs) for the association between recent use of ampicillin/amoxicillin and KPLA. Ampicillin administration predisposed K. pneumoniae-colonized mice to increased bacterial burden, liver abscess and necrosis, and lethality. The population-based study showed that the adjusted OR associating the use of ampicillin/amoxicillin within the past 30 days with KPLA was 3.5 (95% confidence interval, 2.5-5.1). No association was found with use in the past 31-90 days. Ampicillin/amoxicillin therapy started within the past 30 days was associated with increased risk for KPLA. We should avoid the overuse of these antibiotics to prevent undesired disease in the endemic area.

  8. Biosynthesis of poly(3-hydroxypropionate) from glycerol using engineered Klebsiella pneumoniae strain without vitamin B12

    PubMed Central

    Feng, Xinjun; Xian, Mo; Liu, Wei; Xu, Chao; Zhang, Haibo; Zhao, Guang

    2015-01-01

    Poly(3-hydroxypropionate) (P3HP) is a biodegradable and biocompatible thermoplastic. Previous studies demonstrated that engineered Escherichia coli strains can produce P3HP with supplementation of expensive vitamin B12. The present study examined the production of P3HP from glycerol in the recombinant Klebsiella pneumoniae strain, which naturally synthesizes vitamin B12. The genes glycerol dehydratase and its reactivation factor (dhaB123, gdrA, and gdrB from K. pneumoniae), aldehyde dehydrogenase (aldH from E. coli) were cloned and expressed in K. pneumoniae to produce 3-hydroxypropionate (3HP), with 2 genes (dhaT and yqhD) for biosynthesis of 1,3-propanediol were deleted. To obtain P3HP production, propionyl-CoA synthetase (prpE from E. coli) and polyhydroxyalkanoate synthase (phaC from Ralstonia eutropha) were introduced. Under the appropriate aeration condition, the cell yield and P3HP content were 0.24 g/L and 12.7% (wt/wt [cell dry weight]) respectively along with 2.03 g/L 3HP after 48 h cultivation. Although the yield is relatively low, this study shows the feasibility of producing P3HP in K. pneumoniae from glycerol without vitamin B12 for the first time. The results also suggest that the aeration conditions should be optimized carefully for the efficient production of P3HP. PMID:25621933

  9. A Klebsiella pneumoniae antibiotic resistance mechanism that subdues host defences and promotes virulence.

    PubMed

    Kidd, Timothy J; Mills, Grant; Sá-Pessoa, Joana; Dumigan, Amy; Frank, Christian G; Insua, José L; Ingram, Rebecca; Hobley, Laura; Bengoechea, José A

    2017-04-01

    Klebsiella pneumoniae is an important cause of multidrug-resistant infections worldwide. Recent studies highlight the emergence of multidrug-resistant K. pneumoniae strains which show resistance to colistin, a last-line antibiotic, arising from mutational inactivation of the mgrB regulatory gene. However, the precise molecular resistance mechanisms of mgrB-associated colistin resistance and its impact on virulence remain unclear. Here, we constructed an mgrB gene K. pneumoniae mutant and performed characterisation of its lipid A structure, polymyxin and antimicrobial peptide resistance, virulence and inflammatory responses upon infection. Our data reveal that mgrB mutation induces PhoPQ-governed lipid A remodelling which confers not only resistance to polymyxins, but also enhances K. pneumoniae virulence by decreasing antimicrobial peptide susceptibility and attenuating early host defence response activation. Overall, our findings have important implications for patient management and antimicrobial stewardship, while also stressing antibiotic resistance development is not inexorably linked with subdued bacterial fitness and virulence.

  10. N-Acyl Homoserine Lactone Production by Klebsiella pneumoniae Isolated from Human Tongue Surface

    PubMed Central

    Yin, Wai-Fong; Purmal, Kathiravan; Chin, Shenyang; Chan, Xin-Yue; Koh, Chong-Lek; Sam, Choon-Kook; Chan, Kok-Gan

    2012-01-01

    Bacteria communicate by producing quorum sensing molecules called autoinducers, which include autoinducer-1, an N-hexanoyl homoserine lactone (AHL), and autoinducer-2. Bacteria present in the human oral cavity have been shown to produce autoinducer-2, but not AHL. Here, we report the isolation of two AHL-producing Klebsiella pneumoniae strains from the posterior dorsal surface of the tongue of a healthy individual. Spent culture supernatant extracts from K. pneumoniae activated the biosensors Agrobacterium tumefaciens NTL4(pZLR4) and Escherichia coli [pSB401], suggesting the presence of both long and short chain AHLs. High resolution mass spectrometry analyses of these extracts confirmed that both K. pneumoniae isolates produced N-octanoylhomoserine lactone and N-3-dodecanoyl-l-homoserine lactone. To the best of our knowledge, this is the first report of the isolation of K. pneumoniae from the posterior dorsal surface of the human tongue and the production of these AHLs by this bacterium. PMID:22737019

  11. Structure of the core oligosaccharide in the serotype O8 lipopolysaccharide from Klebsiella pneumoniae.

    PubMed Central

    Severn, W B; Kelly, R F; Richards, J C; Whitfield, C

    1996-01-01

    Two classes of mutants with O-antigen-deficient lipopolysaccharides were isolated from the serotype O8 reference strain, belonging to Klebsiella pneumoniae subspecies ozaenae. These mutants were selected by resistance to bacteriophage KO1-2, which recognizes and lyses strains with lipopolysaccharide molecules containing the D-galactan II O antigen. Strain RFK-11 contains a defect in O-antigen synthesis and has a complete core, including the attachment site for O antigen. This mutation is complemented by a plasmid carrying the rfb (O-antigen biosynthesis) gene cluster from the related K. pneumoniae serotype O1. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the lipopolysaccharide from strain RFK-9 has a mobility typical of deep-rough lipopolysaccharide. RFK-9 lipopolysaccharide lacks the attachment site for O antigen. Lipopolysaccharides from strains RFK-9 and RFK-11 were isolated, and their structures were determined by methylation analyses, muclear magnetic resonance spectroscopy, and mass spectroscopy. The deduced O8 core oligosaccharide includes the partial core structure reported for the K. pneumoniae subspecies pneumoniae serotype O1 lipopolysaccharide (M. Süsskind, S. Müller-Leonnies, W. Nimmich, H. Brade, and O. Holst, Carbohydr. Res. 269:C1-7, 1995), consistent with the possibility of a conserved core structure within the species. The core oligosaccharide differs from those of the genera Salmonella and Escherichia by the absence of a hexose-containing outer core, the lack of phosphate residues in the inner core, and the presence of galacturonic acid residues. PMID:8626303

  12. Mast Cell IL-6 Improves Survival From Klebsiella Pneumonia and Sepsis by Enhancing Neutrophil Killing

    PubMed Central

    Sutherland, Rachel E.; Olsen, Joanna S.; McKinstry, Andrew; Villalta, S. Armando; Wolters, Paul J.

    2008-01-01

    The pleiotropic cytokine interleukin 6 (IL-6) has favorable and harmful effects on survival from bacterial infections. While many innate immune cells produce IL-6, little is known about relevant sources in vivo and the nature of its contributions to host responses to severe bacterial infections. To examine these roles, we subjected mast cell-specific IL-6-deficient mice to the cecal ligation and puncture model of septic peritonitis, finding that survival in these mice is markedly worse than in controls. Following intranasal or intraperitoneal inoculation with Klebsiella pneumoniae, IL-6-/- mice are less likely to survive than wild-type controls and at the time of death have higher numbers of bacteria but not inflammatory cells in lungs and peritoneum. Similarly, mast cell-specific IL-6-deficient mice have diminished survival and higher numbers of K. pneumoniae following intraperitoneal infection. Neutrophils lacking IL-6 have greater numbers of live intracellular K. pneumonia, suggesting impaired intracellular killing contributes to reduced clearance in IL-6-/- mice. These results establish that mast cell IL-6 is a critical mediator of survival following K. pneumoniae infection and sepsis and suggest that IL-6 protects from death by augmenting neutrophil killing of bacteria. PMID:18832718

  13. The Antimicrobial Susceptibility of Klebsiella pneumoniae from Community Settings in Taiwan, a Trend Analysis

    PubMed Central

    Lin, Wu-Pu; Wang, Jann-Tay; Chang, Shan-Chwen; Chang, Feng-Yee; Fung, Chang-Phone; Chuang, Yin-Ching; Chen, Yao-Shen; Shiau, Yih-Ru; Tan, Mei-Chen; Wang, Hui-Ying; Lai, Jui-Fen; Huang, I-Wen; Lauderdale, Tsai-Ling

    2016-01-01

    Drug-resistant Klebsiella pneumoniae, especially extended-spectrum β-lactamase (ESBL)- and/or AmpC β-lactamase-producing strains, is an emerging problem worldwide. However, few data focusing on drug susceptibility of K. pneumoniae from community is available. In this study, we analyzed 1016 K. pneumoniae isolates from outpatients or those visiting emergency rooms collected during 2002–2012 from Taiwan Surveillance of Antimicrobial Resistance program. Significantly decreased susceptibilities to 3rd generation cephalosporins and ciprofloxacin were found during the study period. By 2012, susceptibility to cefotaxime and ciprofloxacin was 83.6% and 81.6%, respectively. The prevalence of ESBL-producers increased from 4.8% in 2002 to 11.9% in 2012 (P = 0.012), while that of AmpC β-lactamase-producers increased from 0% to 9.5% in the same period (P < 0.001). Phylogenic analysis of the ESBL and AmpC-β-lactamase-producers by pulsed-field gel electrophoresis and multi-locus sequence typing revealed wide genetic diversity even among the most common sequence type 11 isolates (33.0%). By multivariate analysis, later study year, elderly, and urine isolates were associated with carriage of ESBL genes, while only urine isolates were associated with carriage of AmpC β-lactamase genes. Further studies are needed to determine which antibiotics are reasonable empirical therapy options for patients presenting with severe sepsis that might be caused by K. pneumoniae. PMID:27824151

  14. Porin Loss Impacts the Host Inflammatory Response to Outer Membrane Vesicles of Klebsiella pneumoniae

    PubMed Central

    Turner, Kelli L.; Cahill, Bethaney K.; Dilello, Sarah K.; Gutel, Dedra; Brunson, Debra N.; Albertí, Sebastián

    2015-01-01

    Antibiotic-resistant strains of Klebsiella pneumoniae often exhibit porin loss. In this study, we investigated how porin loss impacted the composition of secreted outer membrane vesicles as well as their ability to trigger proinflammatory cytokine secretion by macrophages. We hypothesize that porin loss associated with antibiotic resistance will directly impact both the composition of outer membrane vesicles and their interactions with phagocytic cells. Using clonally related clinical isolates of extended-spectrum beta-lactamase (ESBL)-positive Klebsiella pneumoniae with different patterns of porin expression, we demonstrated that altered expression of OmpK35 and OmpK36 results in broad alterations to the protein profile of secreted vesicles. Additionally, the level of OmpA incorporation was elevated in strains lacking a single porin. Porin loss significantly impacted macrophage inflammatory responses to purified vesicles. Outer membrane vesicles lacking both OmpK35 and OmpK36 elicited significantly lower levels of proinflammatory cytokine secretion than vesicles from strains expressing one or both porins. These data demonstrate that antibiotic resistance-associated porin loss has a broad and significant effect on both the composition of outer membrane vesicles and their interactions with phagocytic cells, which may impact bacterial survival and inflammatory reactions in the host. PMID:26666932

  15. Aeromonas punctata derived depolymerase that disrupts the integrity of Klebsiella pneumoniae capsule: optimization of depolymerase production.

    PubMed

    Bansal, Shruti; Soni, Sanjeev Kumar; Harjai, Kusum; Chhibber, Sanjay

    2014-07-01

    Formation of dense, highly hydrated biofilm structures pose a risk for public and environmental health. Extracellular polymeric substances encompassing biofilms offer 1000-fold greater resistance as compared to the planktonic cells. Using enzymes as anti-biofouling agents, will improve penetration of antimicrobials and increase susceptibility of biofilms to components of immune system. The challenge of using enzymes derived from unrelated bacteria for the degradation of capsular matrix of Klebsiella pneumoniae has not been dealt in the past. Thus, statistical optimization was done to enhance depolymerase production by Aeromonas punctata, directed against the exopolysaccharide matrix of Klebsiella pneumoniae B5055, capable of substituting the available phage borne depolymerase enzyme. Optimization via central composite design (CCD) resulted in 16-fold enhancement in depolymerase yield (166.65 µmoles ml(-1)  min(-1) ) over unoptimized medium. Out of the 19 variables, media composition giving maximum expression levels of the enzyme consisted of 1 mg ml(-1) galactose and ammonium chloride, 1.5 mg ml(-1) each of capsular polysaccharide (CPS) and magnesium sulfate. Tryptic peptide analysis of the purified 29 kDa band by Matrix assisted laser desorption ionization-time of flight (MALDI-TOF) showed a high homology with a protein of unknown function from Aeromonas cavaie Ae398. Further improvements in the enzyme can lead to its successful development as prophylactic and/or a therapeutic agent.

  16. Pulmonary surfactant as vehicle for intratracheally instilled tobramycin in mice infected with Klebsiella pneumoniae.

    PubMed Central

    van't Veen, A.; Mouton, J. W.; Gommers, D.; Lachmann, B.

    1996-01-01

    1. The use of pulmonary surfactant has been proposed as a vehicle for antibiotic delivery to the alveolar compartment of the lung. This study investigated survival rates of mice with a respiratory Klebsiella pneumoniae infection treated intratracheally with tobramycin using a natural exogenous surfactant preparation as vehicle. 2. At day 1 after infection, animals were injected intratracheally with 20 microliters of the following solutions: (1) a mixture of surfactant (500 micrograms) and tobramycin (250 micrograms); (2) tobramycin (250 micrograms) alone; (3) surfactant (500 micrograms) alone; and (4) NaHCO3 buffer (control, sham-treatment). A fifth group received no treatment (control). Deaths were registered every 12 h for 8 consecutive days. 3. The results show an increased survival in the group receiving the surfactant-tobramycin mixture compared to the group receiving tobramycin alone (P < 0.05), the group receiving surfactant alone (P < 0.01) and the control groups (P < 0.01). It is concluded that intratracheal instillation of surfactant-tobramycin is superior to tobramycin alone in protecting animals from death due to a respiratory Klebsiella pneumoniae infection. PMID:8937717

  17. 1, 4-alpha-Glucan phosphorylase from Klebsiella pneumoniae purification, subunit structure and amino acid composition.

    PubMed

    Linder, D; Kurz, G; Bender, H; Wallenfels, K

    1976-11-01

    1. A 1,4-alpha-glucan phosphorylase from Klebsiella pneumoniae has been purified about 80-fold with an over-all yield greater than 35%. The purified enzyme has been shown to be homogeneous by gel electrophoresis at different pH-values, by isoelectric focusing, by dodecylsulfate electrophoresis and by ultracentrifugation. 2. The molecular weight of the native enzyme has been determined to be 180 000 by ultra-centrifugation studies, in good agreement with the value of 189 000 estimated by gel permeation chromatography. 3. The enzyme dissociates in the presence of 0.1% dodecylsulfate or 5 M guanidine hydrochloride into polypeptide chains. The molecular weight of these polypeptide chains has been found to be 88 000 by dodecylsulfate polyacrylamide gel electrophoresis and 99 000 by sedimentation equilibrium studies, indicating that the native enzyme is composed of two polypeptide chains. 4. The enzyme contains pyridoxalphosphate with a stoichiometry of two moles per 180 000 g protein, confirming that the 1,4-alpha-glucan phosphorylase from Klebsiella pneumoniae is a dimeric enzyme. 5. The amino acid composition of the enzyme has been determined, and its correspondence to that of 1,4-alpha-glucan phosphorylases from other sources is discussed. 6. The pI of the enzyme has been shown to be 5.3 and its pH-optimum to be about pH 5.9. The enzyme is stable in the range from pH 5.9 to 10.5.

  18. Porin Loss Impacts the Host Inflammatory Response to Outer Membrane Vesicles of Klebsiella pneumoniae.

    PubMed

    Turner, Kelli L; Cahill, Bethaney K; Dilello, Sarah K; Gutel, Dedra; Brunson, Debra N; Albertí, Sebastián; Ellis, Terri N

    2015-12-14

    Antibiotic-resistant strains of Klebsiella pneumoniae often exhibit porin loss. In this study, we investigated how porin loss impacted the composition of secreted outer membrane vesicles as well as their ability to trigger proinflammatory cytokine secretion by macrophages. We hypothesize that porin loss associated with antibiotic resistance will directly impact both the composition of outer membrane vesicles and their interactions with phagocytic cells. Using clonally related clinical isolates of extended-spectrum beta-lactamase (ESBL)-positive Klebsiella pneumoniae with different patterns of porin expression, we demonstrated that altered expression of OmpK35 and OmpK36 results in broad alterations to the protein profile of secreted vesicles. Additionally, the level of OmpA incorporation was elevated in strains lacking a single porin. Porin loss significantly impacted macrophage inflammatory responses to purified vesicles. Outer membrane vesicles lacking both OmpK35 and OmpK36 elicited significantly lower levels of proinflammatory cytokine secretion than vesicles from strains expressing one or both porins. These data demonstrate that antibiotic resistance-associated porin loss has a broad and significant effect on both the composition of outer membrane vesicles and their interactions with phagocytic cells, which may impact bacterial survival and inflammatory reactions in the host. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  19. Identifying the shared metabolic objectives of glycerol bioconversion in Klebsiella pneumoniae under different culture conditions.

    PubMed

    Xu, Gongxian; Li, Caixia

    2017-03-18

    This paper addresses the problem of identifying the shared metabolic objectives of glycerol bioconversion in Klebsiella pneumoniae for production of 1,3-propanediol (1,3-PD) under different culture conditions. To achieve this goal, we propose a multi-level programming model. This model includes three optimization problems, where the constraint region of the first level problem is implicitly determined by the other two optimization problems. The optimized objectives of the first and second level problems are to minimize the set of fluxes that are of major importance to glycerol metabolism and the difference between the observed fluxes and those computed by the model, respectively. The third level problem in the proposed multi-level programming simultaneously solves a set of flux balance analysis (FBA) models. A method is proposed to solve efficiently the presented multi-level programming problem. In this method, we first transform the proposed multi-level problem into a bi-level problem by applying the dual theory of linear programming to the FBA models of the third level. Next, the optimal solution of the above bi-level problem is obtained by iteratively solving a sequence of mixed integer programming problems. Optimization results reveal that the proposed method can identify the shared metabolic objectives of glycerol bioconversion in Klebsiella pneumoniae under three groups of experimental data.

  20. Dipeptidyl peptidase I controls survival from Klebsiella pneumoniae lung infection by processing surfactant protein D.

    PubMed

    Sutherland, Rachel E; Barry, Sophia S; Olsen, Joanna S; Salantes, D Brenda; Caughey, George H; Wolters, Paul J

    2014-07-18

    Prior work established that a deficiency in the cysteine protease dipeptidyl peptidase I (DPPI) improves survival following polymicrobial septic peritonitis. To test whether DPPI regulates survival from severe lung infections, DPPI(-/-) mice were studied in a Klebsiella pneumoniae lung infection model, finding that survival in DPPI(-/-) mice is significantly better than in DPPI(+/+) mice 8d after infection. DPPI(-/-) mice have significantly fewer bacteria in the lung than infected DPPI(+/+) mice, but no difference in lung histopathology, lung injury, or cytokine levels. To explore mechanisms of enhanced bacterial clearance in DPPI(-/-) mice, we examined the status of pulmonary collectins, finding that levels of surfactant protein D, but not of surfactant protein A, are higher in DPPI(-/-) than in DPPI(+/+) BAL fluid, and that DPPI(-/-) BAL fluid aggregate bacteria more effectively than control BAL fluid. Sequencing of the amino terminus of surfactant protein D revealed two or eight additional amino acids in surfactant protein D isolated from DPPI(-/-) mice, suggesting processing by DPPI. These results establish that DPPI is a major determinant of survival following Klebsiella pneumoniae lung infection and suggest that the survival disadvantage in DPPI(+/+) mice is in part due to processing of surfactant protein D by DPPI. Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Dipeptidyl Peptidase I Controls Survival from Klebsiella pneumoniae Lung Infection by Processing Surfactant Protein D 1

    PubMed Central

    Olsen, Joanna S.; Salantes, D. Brenda; Caughey, George H.; Wolters, Paul J.

    2014-01-01

    Prior work established that a deficiency in the cysteine protease dipeptidyl peptidase I (DPPI) improves survival following polymicrobial septic peritonitis. To test whether DPPI regulates survival from severe lung infections, DPPI −/− mice were studied in a Klebsiella pneumonia lung infection model, finding that survival in DPPI −/− mice is significantly better than in DPPI +/+ mice 8 d after infection. DPPI −/− mice have significantly fewer bacteria in the lung than infected DPPI +/+ mice, but no difference in lung histopathology, lung injury, or cytokine levels. To explore mechanisms of enhanced bacterial clearance in DPPI −/− mice, we examined the status of pulmonary collectins, finding that levels of surfactant protein D, but not of surfactant protein A, are higher in DPPI −/− than in DPPI +/+ BAL fluid, and that DPPI −/− BAL fluid aggregate bacteria more effectively than control BAL fluid. Sequencing of the amino terminus of surfactant protein D revealed two or eight additional amino acids in surfactant protein D isolated from DPPI −/− mice, suggesting processing by DPPI. These results establish that DPPI is a major determinant of survival following Klebsiella pneumoniae lung infection and suggest that the survival disadvantage in DPPI +/+ mice is in part due to processing of surfactant protein D by DPPI. PMID:24955853

  2. Sublethal Concentrations of Carbapenems Alter Cell Morphology and Genomic Expression of Klebsiella pneumoniae Biofilms

    PubMed Central

    Van Laar, Tricia A.; Chen, Tsute; You, Tao

    2015-01-01

    Klebsiella pneumoniae, a Gram-negative bacterium, is normally associated with pneumonia in patients with weakened immune systems. However, it is also a prevalent nosocomial infectious agent that can be found in infected surgical sites and combat wounds. Many of these clinical strains display multidrug resistance. We have worked with a clinical strain of K. pneumoniae that was initially isolated from a wound of an injured soldier. This strain demonstrated resistance to many commonly used antibiotics but sensitivity to carbapenems. This isolate was capable of forming biofilms in vitro, contributing to its increased antibiotic resistance and impaired clearance. We were interested in determining how sublethal concentrations of carbapenem treatment specifically affect K. pneumoniae biofilms both in morphology and in genomic expression. Scanning electron microscopy showed striking morphological differences between untreated and treated biofilms, including rounding, blebbing, and dimpling of treated cells. Comparative transcriptome analysis using RNA sequencing (RNA-Seq) technology identified a large number of open reading frames (ORFs) differentially regulated in response to carbapenem treatment at 2 and 24 h. ORFs upregulated with carbapenem treatment included genes involved in resistance, as well as those coding for antiporters and autoinducers. ORFs downregulated included those coding for metal transporters, membrane biosynthesis proteins, and motility proteins. Quantitative real-time PCR validated the general trend of some of these differentially regulated ORFs. Treatment of K. pneumoniae biofilms with sublethal concentrations of carbapenems induced a wide range of phenotypic and gene expression changes. This study reveals some of the mechanisms underlying how sublethal amounts of carbapenems could affect the overall fitness and pathogenic potential of K. pneumoniae biofilm cells. PMID:25583711

  3. Genome-Wide Identification of Klebsiella pneumoniae Fitness Genes during Lung Infection

    PubMed Central

    Breen, Paul; Deornellas, Valerie; Mu, Qiao; Zhao, Lili; Wu, Weisheng; Cavalcoli, James D.; Mobley, Harry L. T.

    2015-01-01

    ABSTRACT Klebsiella pneumoniae is an urgent public health threat because of resistance to carbapenems, antibiotics of last resort against Gram-negative bacterial infections. Despite the fact that K. pneumoniae is a leading cause of pneumonia in hospitalized patients, the bacterial factors required to cause disease are poorly understood. Insertion site sequencing combines transposon mutagenesis with high-throughput sequencing to simultaneously screen thousands of insertion mutants for fitness defects during infection. Using the recently sequenced K. pneumoniae strain KPPR1 in a well-established mouse model of pneumonia, insertion site sequencing was performed on a pool of >25,000 transposon mutants. The relative fitness requirement of each gene was ranked based on the ratio of lung to inoculum read counts and concordance between insertions in the same gene. This analysis revealed over 300 mutants with at least a 2-fold fitness defect and 69 with defects ranging from 10- to >2,000-fold. Construction of 6 isogenic mutants for use in competitive infections with the wild type confirmed their requirement for lung fitness. Critical fitness genes included those for the synthesis of branched-chain and aromatic amino acids that are essential in mice and humans, the transcriptional elongation factor RfaH, and the copper efflux pump CopA. The majority of fitness genes were conserved among reference strains representative of diverse pathotypes. These results indicate that regulation of outer membrane components and synthesis of amino acids that are essential to its host are critical for K. pneumoniae fitness in the lung. PMID:26060277

  4. Carbapenemases-producing Klebsiella pneumoniae in hospitals of two regions of Southern Italy.

    PubMed

    Calia, Carla; Pazzani, Carlo; Oliva, Marta; Scrascia, Maria; Lovreglio, Piero; Capolongo, Carmen; Dionisi, Anna Maria; Chiarelli, Adriana; Monno, Rosa

    2017-05-01

    Carbapenem-resistant Klebsiella pneumoniae infections are reported with increasing frequency elsewhere in the world, representing a worrying phenomenon for global health. In Italy, there are hotspot data on the diffusion and type of carbapenemase-producing Enterobacteriaceae and K. pneumoniae in particular, with very few data coming from Apulia and Basilicata, two regions of Southern Italy. This study was aimed at characterizing by phenotypic and genotypic methods carbapenem-resistant K. pneumoniae isolated from several Hospitals of Apulia and Basilicata, Southern Italy. Antibiotic susceptibility was also evaluated. The relatedness of carbapenemase-producing K. pneumoniae strains was established by pulsed-field gel electrophoresis (PFGE). Among the 150 K. pneumoniae carbapenemase producers, KPC-3 genotype was the most predominant (95%), followed by VIM-1 (5%). No other genotypes were found and no co-presence of two carbapenemase genes was found. A full concordance between results obtained by both the phenotypic and the genotypic tests was observed. All strains were resistant to β-lactam antibiotics including carbapenems, and among antibiotics tested, only tetracycline and gentamycin showed low percentage of resistance (18% and 15%, respectively). Resistance to colistin was detected in 17.3% of strains studied. The analysis of PFGE profiles of the carbapenemases-positive strains shows that one group (B) of the five (A to E) main groups identified was the most prevalent and detected in almost all the hospitals considered, while the other groups were randomly distributed. Three different sequence types (ST 307, ST 258, and ST 512) were detected with the majority of isolates belonging to the ST 512. Our results demonstrated the wide diffusion of K. pneumoniae KPC-3 in the area considered, the good concordance between phenotypic and genotypic tests. Gentamicin and colistin had a good activity against these strains. © 2017 APMIS. Published by John Wiley & Sons

  5. Multi-functional analysis of Klebsiella pneumoniae fimbrial types in adherence and biofilm formation.

    PubMed

    Alcántar-Curiel, María D; Blackburn, Dana; Saldaña, Zeus; Gayosso-Vázquez, Catalina; Iovine, Nicole M; De la Cruz, Miguel A; Girón, Jorge A

    2013-02-15

    Klebsiella pneumoniae is an opportunistic pathogen frequently associated with nosocomially acquired infections. Host cell adherence and biofilm formation of K. pneumoniae isolates is mediated by type 1 (T1P) and type 3 (MR/K) pili whose major fimbrial subunits are encoded by the fimA and mrkA genes, respectively. The E. coli common pilus (ECP) is an adhesive structure produced by all E. coli pathogroups and a homolog of the ecpABCDE operon is present in the K. pneumoniae genome. In this study, we aimed to determine the prevalence of these three fimbrial genes among a collection of 69 clinical and environmental K. pneumoniae strains and to establish a correlation with fimbrial production during cell adherence and biofilm formation. The PCR-based survey demonstrated that 96% of the K. pneumoniae strains contained ecpA and 94% of these strains produced ECP during adhesion to cultured epithelial cells. Eighty percent of the strains forming biofilms on glass produced ECP, suggesting that ECP is required, at least in vitro, for expression of these phenotypes. The fim operon was found in 100% of the strains and T1P was detected in 96% of these strains. While all the strains examined contained mrkA, only 57% of them produced MR/K fimbriae, alone or together with ECP. In summary, this study highlights the ability of K. pneumoniae strains to produce ECP, which may represent a new important adhesive structure of this organism. Further, it defines the multi-fimbrial nature of the interaction of this nosocomial pathogen with host epithelial cells and inert surfaces.

  6. Evaluation of Biofilm Formation Among Klebsiella pneumoniae Isolates and Molecular Characterization by ERIC-PCR

    PubMed Central

    Seifi, Kimia; Kazemian, Hossein; Heidari, Hamid; Rezagholizadeh, Fereshteh; Saee, Yasaman; Shirvani, Fariba; Houri, Hamidreza

    2016-01-01

    Background: Klebsiella pneumoniae is among the most frequently recovered etiologic agents from nosocomial infections. This opportunistic pathogen can generate a thick layer of biofilm as one of its important virulence factors, enabling the bacteria to attach to living or abiotic surfaces, which contributes to drug resistance. Objectives: The resistance of biofilm-mediated infections to effective chemotherapy has adverse effects on patient outcomes and survival. Therefore, the aim of the present study was to evaluate the biofilm-formation capacity of clinical K. pneumoniae isolates and to perform a molecular characterization using enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) to determine the dominant biofilm-producing genotype. Patients and Methods: In the present study, 94 K. pneumoniae isolates were obtained from two hospitals in Tehran, Iran. Biofilm formation was assayed by a modified procedure, then ERIC-PCR was carried out. Results: The distributions of the clinical specimens used in this study were 61.7% from urine, 18.1% from wounds, 11.7% from sputum, and 8.5% from blood. Among these isolates, 33% formed fully established biofilms, 52.1% were categorized as moderately biofilm-producing, 8.5% formed weak biofilms, and 6.4% were non-biofilm-producers. Genotyping of K. pneumoniae revealed 31 different ERIC types. Biofilm-formation ability in a special ERIC type was not observed. Conclusions: Our results indicated that an enormous proportion of K. pneumoniae isolated from sputum and surgical-wound swabs produced fully established biofilms. It is reasonable to assume the existence of a relationship between the site of infection and the formation of biofilm. A high level of genetic diversity among the K. pneumoniae strains was observed. PMID:27099694

  7. Sublethal concentrations of carbapenems alter cell morphology and genomic expression of Klebsiella pneumoniae biofilms.

    PubMed

    Van Laar, Tricia A; Chen, Tsute; You, Tao; Leung, Kai P

    2015-03-01

    Klebsiella pneumoniae, a Gram-negative bacterium, is normally associated with pneumonia in patients with weakened immune systems. However, it is also a prevalent nosocomial infectious agent that can be found in infected surgical sites and combat wounds. Many of these clinical strains display multidrug resistance. We have worked with a clinical strain of K. pneumoniae that was initially isolated from a wound of an injured soldier. This strain demonstrated resistance to many commonly used antibiotics but sensitivity to carbapenems. This isolate was capable of forming biofilms in vitro, contributing to its increased antibiotic resistance and impaired clearance. We were interested in determining how sublethal concentrations of carbapenem treatment specifically affect K. pneumoniae biofilms both in morphology and in genomic expression. Scanning electron microscopy showed striking morphological differences between untreated and treated biofilms, including rounding, blebbing, and dimpling of treated cells. Comparative transcriptome analysis using RNA sequencing (RNA-Seq) technology identified a large number of open reading frames (ORFs) differentially regulated in response to carbapenem treatment at 2 and 24 h. ORFs upregulated with carbapenem treatment included genes involved in resistance, as well as those coding for antiporters and autoinducers. ORFs downregulated included those coding for metal transporters, membrane biosynthesis proteins, and motility proteins. Quantitative real-time PCR validated the general trend of some of these differentially regulated ORFs. Treatment of K. pneumoniae biofilms with sublethal concentrations of carbapenems induced a wide range of phenotypic and gene expression changes. This study reveals some of the mechanisms underlying how sublethal amounts of carbapenems could affect the overall fitness and pathogenic potential of K. pneumoniae biofilm cells.

  8. Identification of Klebsiella pneumoniae virulence determinants using an intranasal infection model.

    PubMed

    Lawlor, Matthew S; Hsu, James; Rick, Paul D; Miller, Virginia L

    2005-11-01

    Klebsiella pneumoniae is a Gram-negative enterobacterium that has historically been, and currently remains, a significant cause of human disease. It is a frequent cause of urinary tract infections and pneumonia, and subsequent systemic infections can have mortality rates as high as 60%. Despite its clinical significance, few virulence factors of K. pneumoniae have been identified or characterized. In this study we present a mouse model of acute K. pneumoniae respiratory infection using an intranasal inoculation method, and examine the progression of both pulmonary and systemic disease. Wild-type infection recapitulates many aspects of clinical disease, including significant bacterial growth in both the trachea and lungs, an inflammatory immune response characterized by dramatic neutrophil influx, and a steady progression to systemic disease with ensuing mortality. These observations are contrasted with an infection by an isogenic capsule-deficient strain that shows an inability to cause disease in either pulmonary or systemic tissues. The consistency and clinical accuracy of the intranasal mouse model proved to be a useful tool as we conducted a genetic screen to identify novel virulence factors of K. pneumoniae. A total of 4800 independent insertional mutants were evaluated using a signature-tagged mutagenesis protocol. A total of 106 independent mutants failed to be recovered from either the lungs or spleens of infected mice. Small scale independent infections proved to be helpful as a secondary screening method, as opposed to the more traditional competitive index assay. Those mutants showing verified attenuation contained insertions in loci with a variety of putative functions, including a large number of hypothetical open reading frames. Subsequent experiments support the premise that the central mechanism of K. pneumoniae pathogenesis is the production of a polysaccharide-rich cell surface that provides protection from the inflammatory response.

  9. Emergence of New Delhi Metallo-Beta-Lactamase (NDM-1) and Klebsiella pneumoniae Carbapenemase (KPC-2) in South Africa

    PubMed Central

    Coetzee, Jennifer; Clay, Cornelis G.; Sithole, Sindi; Richards, Guy A.; Poirel, Laurent; Nordmann, Patrice

    2012-01-01

    This report documents emergence of New Delhi metallo-beta-lactamase (NDM-1) and Klebsiella pneumoniae carbapenemase (KPC-2) in K. pneumoniae and Enterobacter cloacae in South Africa. NDM-1 producers have not been described in South Africa, and this is the first instance that KPC producers have been identified in Africa. The two patients infected with these carbapenemase-producing bacteria demised. PMID:22116157

  10. Emergence of KPC-producing Klebsiella pneumoniae ST512 isolated from cerebrospinal fluid of a child in Algeria

    PubMed Central

    Bakour, S.; Sahli, F.; Touati, A.; Rolain, J.-M.

    2014-01-01

    We report class A carbapenemase (KPC)-3-producing Klebsiella pneumoniae meningitis in a 6-month-old child in Algeria. Multilocus sequence typing showed that the sequence type obtained corresponded to ST512, an allelic single-locus variant of the pandemic ST258 widely distributed in KPC producers from Europe. To our knowledge, this is the first report of KPC-3-producing K. pneumoniae ST512 in a North African country. PMID:25755890

  11. Draft Genome Sequence of a Colistin-Resistant Klebsiella pneumoniae Clinical Strain Carrying the blaNDM-1 Carbapenemase Gene

    PubMed Central

    Yao, Zhihong; Feng, Yu; Lin, Ji

    2017-01-01

    ABSTRACT Klebsiella pneumoniae strain WCHKP1845, recovered from the sputum of a patient with pneumonia, was resistant to colistin and carried the carbapenemase gene blaNDM-1. Here, we report its 5.4-Mb draft genome sequence, comprising 140 contigs with an average 57.33% G+C content. The genome contains 5,118 coding sequences and 88 tRNA genes. PMID:28209835

  12. In-vivo study of the nuclear quadrupole interaction of99Mo (β- 99)Tc in nitrogenase of Klebsiella pneumoniaein nitrogenase of Klebsiella pneumoniae

    NASA Astrophysics Data System (ADS)

    Mottner, P.; Lerf, A.; Ni, X.; Butz, T.; Erfkamp, J.; Müller, A.

    1990-08-01

    We report on the first TDPAC-measurements of the nuclear quadrupole interaction (NQI) of (NQI) of99Mo(β-)99Tc in the nitrogenase of the bacteria Klebsiella pneumoniae. Because nitrogenase is the only Mo-containing enzyme in Klebsiella pneumoniae under the chosen conditions, no further isolation of this enzyme was necessary. The majority of the incorporated99Mo is subjected to a well defined NQI with ω=365(7) Mrad/s, η=1 and a reorientational correlation time of τcoττ≈10nsec and is attributed to the active site of the FeMo cofactor. During sample preparation we noted a pronounced affinity of the bacteria to99mTc.

  13. Klebsiella pneumoniae Carbapenemase (KPC)-Producing K. pneumoniae at a Single Institution: Insights into Endemicity from Whole-Genome Sequencing

    PubMed Central

    Stoesser, Nicole; Sheppard, Anna E.; Pankhurst, Louise; Giess, Adam; Yeh, Anthony J.; Didelot, Xavier; Turner, Stephen D.; Sebra, Robert; Kasarskis, Andrew; Peto, Tim; Crook, Derrick; Sifri, Costi D.

    2015-01-01

    The global emergence of Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC-Kp) multilocus sequence type ST258 is widely recognized. Less is known about the molecular and epidemiological details of non-ST258 K. pneumoniae in the setting of an outbreak mediated by an endemic plasmid. We describe the interplay of blaKPC plasmids and K. pneumoniae strains and their relationship to the location of acquisition in a U.S. health care institution. Whole-genome sequencing (WGS) analysis was applied to KPC-Kp clinical isolates collected from a single institution over 5 years following the introduction of blaKPC in August 2007, as well as two plasmid transformants. KPC-Kp from 37 patients yielded 16 distinct sequence types (STs). Two novel conjugative blaKPC plasmids (pKPC_UVA01 and pKPC_UVA02), carried by the hospital index case, accounted for the presence of blaKPC in 21/37 (57%) subsequent cases. Thirteen (35%) isolates represented an emergent lineage, ST941, which contained pKPC_UVA01 in 5/13 (38%) and pKPC_UVA02 in 6/13 (46%) cases. Seven (19%) isolates were the epidemic KPC-Kp strain, ST258, mostly imported from elsewhere and not carrying pKPC_UVA01 or pKPC_UVA02. Using WGS-based analysis of clinical isolates and plasmid transformants, we demonstrate the unexpected dispersal of blaKPC to many non-ST258 lineages in a hospital through spread of at least two novel blaKPC plasmids. In contrast, ST258 KPC-Kp was imported into the institution on numerous occasions, with other blaKPC plasmid vectors and without sustained transmission. Instead, a newly recognized KPC-Kp strain, ST941, became associated with both novel blaKPC plasmids and spread locally, making it a future candidate for clinical persistence and dissemination. PMID:25561339

  14. Genomic Epidemiology of an Endoscope-Associated Outbreak of Klebsiella pneumoniae Carbapenemase (KPC)-Producing K. pneumoniae

    PubMed Central

    Marsh, Jane W.; Krauland, Mary G.; Nelson, Jemma S.; Schlackman, Jessica L.; Brooks, Anthony M.; Pasculle, A. William; Shutt, Kathleen A.; Doi, Yohei; Querry, Ashley M.; Muto, Carlene A.; Harrison, Lee H.

    2015-01-01

    Increased incidence of infections due to Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae (KPC-Kp) was noted among patients undergoing endoscopic retrograde cholangiopancreatography (ERCP) at a single hospital. An epidemiologic investigation identified KPC-Kp and non-KPC-producing, extended-spectrum β-lactamase (ESBL)-producing Kp in cultures from 2 endoscopes. Genotyping was performed on patient and endoscope isolates to characterize the microbial genomics of the outbreak. Genetic similarity of 51 Kp isolates from 37 patients and 3 endoscopes was assessed by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Five patient and 2 endoscope isolates underwent whole genome sequencing (WGS). Two KPC-encoding plasmids were characterized by single molecule, real-time sequencing. Plasmid diversity was assessed by endonuclease digestion. Genomic and epidemiologic data were used in conjunction to investigate the outbreak source. Two clusters of Kp patient isolates were genetically related to endoscope isolates by PFGE. A subset of patient isolates were collected post-ERCP, suggesting ERCP endoscopes as a possible source. A phylogeny of 7 Kp genomes from patient and endoscope isolates supported ERCP as a potential source of transmission. Differences in gene content defined 5 ST258 subclades and identified 2 of the subclades as outbreak-associated. A novel KPC-encoding plasmid, pKp28 helped define and track one endoscope-associated ST258 subclade. WGS demonstrated high genetic relatedness of patient and ERCP endoscope isolates suggesting ERCP-associated transmission of ST258 KPC-Kp. Gene and plasmid content discriminated the outbreak from endemic ST258 populations and assisted with the molecular epidemiologic investigation of an extended KPC-Kp outbreak. PMID:26637170

  15. Genomic Epidemiology of an Endoscope-Associated Outbreak of Klebsiella pneumoniae Carbapenemase (KPC)-Producing K. pneumoniae.

    PubMed

    Marsh, Jane W; Krauland, Mary G; Nelson, Jemma S; Schlackman, Jessica L; Brooks, Anthony M; Pasculle, A William; Shutt, Kathleen A; Doi, Yohei; Querry, Ashley M; Muto, Carlene A; Harrison, Lee H

    2015-01-01

    Increased incidence of infections due to Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae (KPC-Kp) was noted among patients undergoing endoscopic retrograde cholangiopancreatography (ERCP) at a single hospital. An epidemiologic investigation identified KPC-Kp and non-KPC-producing, extended-spectrum β-lactamase (ESBL)-producing Kp in cultures from 2 endoscopes. Genotyping was performed on patient and endoscope isolates to characterize the microbial genomics of the outbreak. Genetic similarity of 51 Kp isolates from 37 patients and 3 endoscopes was assessed by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Five patient and 2 endoscope isolates underwent whole genome sequencing (WGS). Two KPC-encoding plasmids were characterized by single molecule, real-time sequencing. Plasmid diversity was assessed by endonuclease digestion. Genomic and epidemiologic data were used in conjunction to investigate the outbreak source. Two clusters of Kp patient isolates were genetically related to endoscope isolates by PFGE. A subset of patient isolates were collected post-ERCP, suggesting ERCP endoscopes as a possible source. A phylogeny of 7 Kp genomes from patient and endoscope isolates supported ERCP as a potential source of transmission. Differences in gene content defined 5 ST258 subclades and identified 2 of the subclades as outbreak-associated. A novel KPC-encoding plasmid, pKp28 helped define and track one endoscope-associated ST258 subclade. WGS demonstrated high genetic relatedness of patient and ERCP endoscope isolates suggesting ERCP-associated transmission of ST258 KPC-Kp. Gene and plasmid content discriminated the outbreak from endemic ST258 populations and assisted with the molecular epidemiologic investigation of an extended KPC-Kp outbreak.

  16. Successful treatment of pan-resistant Klebsiella pneumoniae pneumonia and bacteraemia with a combination of high-dose tigecycline and colistin.

    PubMed

    Humphries, Romney M; Kelesidis, Theodoros; Dien Bard, Jennifer; Ward, Kevin W; Bhattacharya, Debika; Lewinski, Michael A

    2010-11-01

    The spread of antimicrobial resistance among members of the Enterobacteriaceae is a significant clinical threat. We report the treatment of pan-resistant Klebsiella pneumoniae bacteraemia with combination tigecycline and colistin in a 49-year-old male and review available therapeutic options. Despite a poor prognosis, the patient recovered, but remains colonized with the pan-resistant isolate.

  17. Anti-Biofilm Activity: A Function of Klebsiella pneumoniae Capsular Polysaccharide

    PubMed Central

    Dos Santos Goncalves, Marina; Delattre, Cédric; Balestrino, Damien; Charbonnel, Nicolas; Elboutachfaiti, Redouan; Wadouachi, Anne; Badel, Stéphanie; Bernardi, Thierry; Michaud, Philippe; Forestier, Christiane

    2014-01-01

    Competition and cooperation phenomena occur within highly interactive biofilm communities and several non-biocides molecules produced by microorganisms have been described as impairing biofilm formation. In this study, we investigated the anti-biofilm capacities of an ubiquitous and biofilm producing bacterium, Klebsiella pneumoniae. Cell-free supernatant from K. pneumoniae planktonic cultures showed anti-biofilm effects on most Gram positive bacteria tested but also encompassed some Gram negative bacilli. The anti-biofilm non-bactericidal activity was further investigated on Staphylococcus epidermidis, by determining the biofilm biomass, microscopic observations and agglutination measurement through a magnetic bead-mediated agglutination test. Cell-free extracts from K. pneumoniae biofilm (supernatant and acellular matrix) also showed an influence, although to a lesser extend. Chemical analyses indicated that the active molecule was a high molecular weight polysaccharide composed of five monosaccharides: galactose, glucose, rhamnose, glucuronic acid and glucosamine and the main following sugar linkage residues [→2)-α-l-Rhap-(1→]; [→4)-α-l-Rhap-(1→]; [α-d-Galp-(1→]; [→2,3)-α-d-Galp-(1→]; [→3)-β-d-Galp-(1→] and, [→4)-β-d-GlcAp-(1→]. Characterization of this molecule indicated that this component was more likely capsular polysaccharide (CPS) and precoating of abiotic surfaces with CPS extracts from different serotypes impaired the bacteria-surface interactions. Thus the CPS of Klebsiella would exhibit a pleiotropic activity during biofilm formation, both stimulating the initial adhesion and maturation steps as previously described, but also repelling potential competitors. PMID:24932475

  18. Carbapenem-resistant Klebsiella pneumoniae harbouring blaKPC-3 and blaVIM-2 from central Italy.

    PubMed

    Perilli, Mariagrazia; Bottoni, Carlo; Grimaldi, Alessandro; Segatore, Bernardetta; Celenza, Giuseppe; Mariani, Maurizio; Bellio, Pierangelo; Frascaria, Patrizia; Amicosante, Gianfranco

    2013-02-01

    The frequency of Klebsiella pneumoniae carbapenemase-producing K. pneumoniae is increasing in Italian hospitals and poses an emerging threat to the management of infections in hospitalized patients. In this study, we report a detailed molecular characterization of a K. pneumoniae subsp. pneumoniae KP1/11 isolate from the decubitus ulcer of a hospitalized patient with a serious infection. K. pneumoniae KP1/11 produces KPC-3 and VIM-2 β-lactamases. The bla(KPC-3) gene is harbored in a large plasmid in a complex structure of Tn3-based transposon, Tn4401a. The chromosomal DNA of K. pneumoniae harbored also 2 class 1 integrons with different variable regions: 1) orfD-aacA8; 2) aacA29-bla(VIM-2).

  19. Genome Sequence of Klebsiella pneumoniae KpQ3, a DHA-1 β-Lactamase-Producing Nosocomial Isolate

    PubMed Central

    Tobes, Raquel; Codoñer, Francisco M.; López-Camacho, Elena; Salanueva, Iñigo J.; Manrique, Marina; Brozynska, Marta; Gómez-Gil, Rosa; Martínez-Blanch, Juan F.; Álvarez-Tejado, Miguel; Pareja, Eduardo

    2013-01-01

    Klebsiella pneumoniae KpQ3 is a multidrug-resistant isolate obtained from a blood culture of a patient in a burn unit in the Hospital Universitario La Paz (Madrid, Spain) in 2008. The genome contains multiple antibiotic resistance genes, including a plasmid-mediated DHA-1 cephalosporinase gene. PMID:23469341

  20. Genome Sequence of Klebsiella pneumoniae YZUSK-4, a Bacterium Proposed as a Starter Culture for Fermented Meat Products.

    PubMed

    Yu, Hai; Yin, Yongqi; Xu, Lin; Yan, Ming; Fang, Weiming; Ge, Qingfeng

    2015-07-23

    Klebsiella pneumoniae strain YZUSK-4, isolated from Chinese RuGao ham, is an efficient branched-chain aminotransferase-producing bacterium that can be used widely in fermented meat products to enhance flavor. The draft genome sequence of strain YZUSK-4 may provide useful genetic information on branched-chain amino acid aminotransferase production and branched-chain amino acid metabolism.

  1. Rapid containment of coincident outbreaks with 2 carbapenem-resistant Klebsiella pneumoniae strains in an intensive care unit.

    PubMed

    Meybeck, Agnès; Laurans, Caroline; Broucqsault-Dedrie, Céline; Vanbaelinghem, Clément; Verheyde, Isabelle; Ledez, Rita; Nyunga, Martine; Nottebaert, Sandrine; Waes, Sylvaine; Vachée, Anne

    2014-08-01

    In our intensive care unit, coincident outbreaks were caused by concomitant cross-transmission of 2 carbapenem-resistant Klebsiella pneumoniae strains harboring distinct mechanisms of resistance. One strain produced extended-spectrum ß-lactamase in combination with reduced permeability. The other produced oxacillinase-48 carbapenemase. Rapid phenotypic detection of carbapenemase production allowed timely implementation of appropriate infection control measures.

  2. Genome Sequence of Klebsiella pneumoniae YZUSK-4, a Bacterium Proposed as a Starter Culture for Fermented Meat Products

    PubMed Central

    Yu, Hai; Yin, Yongqi; Yan, Ming; Fang, Weiming; Ge, Qingfeng

    2015-01-01

    Klebsiella pneumoniae strain YZUSK-4, isolated from Chinese RuGao ham, is an efficient branched-chain aminotransferase-producing bacterium that can be used widely in fermented meat products to enhance flavor. The draft genome sequence of strain YZUSK-4 may provide useful genetic information on branched-chain amino acid aminotransferase production and branched-chain amino acid metabolism. PMID:26205853

  3. 1H and 13C NMR characterization and secondary structure of the K2 polysaccharide of Klebsiella pneumoniae strain 52145.

    PubMed

    Corsaro, Maria Michela; De Castro, Cristina; Naldi, Teresa; Parrilli, Michelangelo; Tomás, Juan M; Regué, Miguel

    2005-09-26

    The complete (1)H and (13)C NMR characterization of the tetrasaccharide repeating unit from the K2 polysaccharide of Klebsiella pneumoniae strain 52145 is reported. [chemical structure] In addition a model for its secondary structure was suggested on the basis of dynamic and molecular calculations.

  4. Genetic diversity of genes encoding OKP and LEN beta-lactamases produced by clinical Klebsiella pneumoniae strains in Portugal.

    PubMed

    Mendonça, Nuno; Ferreira, Eugénia; Caniça, Manuela

    2009-03-01

    Of the 308 clinical Klebsiella pneumoniae strains collected in 21 Portuguese health institutions, 11 encoded for LEN and 9 for OKP enzymes; of these, 15 were new enzymes. Ninety-one percent of LEN and all OKP producer strains were resistant to amoxicillin. We demonstrate that these beta-lactamase were highly diverse.

  5. [Molecular typing of Klebsiella pneumonia by pulse-field gel electrophoresis in combination with multilocus sequence typing].

    PubMed

    Gao, Yuan; Zhang, Li; Li, Ma-chao; Huang, Cheng; Ren, Hong-yu; Zhou, Hai-jian; Zhu, Bing-qing; Li, Qian; Wang, Xiao-lei; Shao, Zhu-jun

    2010-07-01

    To type Klebsiella pneumonia through methods including pulse-field gel electrophoresis (PFGE) in combination with multilocus sequence typing. Four selected different EPs, referring to the Standard Operating Procedure of PulseNet China, were used. The single colony of Klebsiella pneumonia was quantified after enriched culture. Embedding organisms in agarose and genome DNA were lysed with Proteinase K and then digested by restriction endonuclease XbaI, to produce agarose gel. Fingerprint was obtained through PFGE and bands were marked with their molecular weights and then analyzed by BioNumerics software. Using MLST to analyze the strains that were highly similar, by PFGE typing By comparing the four results from each EPs, fk3 (switch time from 6s to 36s, total run time is 18.5 hours) seemed to be better than the others. 59 strains of Klebsiella pneumonia were divided into 47 PFGE types and 19 PFGE clusters. The highly similar strains could be typed into ST-340, ST-342, ST-343, ST-344, ST-345 by MLST. Among them, ST-342, ST-343, ST-344, ST-345 types were all new MLST types that were reported in China. Highly similar Klebsiella pneumonias typed by PFGE could also be typed by MLST.

  6. Is multiresistant Klebsiella pneumoniae New Delhi metallo-beta-lactamase (NDM-1) a new threat for kidney transplant recipients?

    PubMed

    Karczewski, M; Tomczak, H; Piechocka-Idasiak, I; Cichanska, L; Adamska, Z; Stronka, M

    2014-09-01

    Urinary tract infections (UTIs) are the most frequent infections among kidney transplant (KT) patients. This case documents the emergence of New Delhi metallo-beta-lactamase (NDM-1) Klebsiella pneumonia--a factor of recurrent post-KT UTI, leading to graft loss. Spreading globally, and multidrug resistant, NDM-1 may become a great threat to transplant patients all over the world.

  7. Enhanced Peroxide Resistance of In Vitro Mutagenized Fluorideresistant Klebsiella pneumoniae Ureases for Catalytic Buffering of Agent Decontamination Reactions

    DTIC Science & Technology

    2004-11-17

    1 ENHANCED PEROXIDE RESISTANCE OF IN VITRO MUTAGENIZED FLUORIDE- RESISTANT Klebsiella pneumoniae UREASES FOR CATALYTIC BUFFERING OF...oxidative surety agent decontamination technologies. Ammonia production from urea by urease neutralizes the production of O- alkylphosphonic acids...resulting from the hydrolysis of Nerve agents such as Sarin and VX. Fluoride production from Sarin hydrolysis inhibits native urease at low mM

  8. Coproduction of KPC-2 and QnrB19 in Klebsiella pneumoniae ST340 isolate in Brazil.

    PubMed

    Martins, Willames M B S; Almeida, Anna C S; Nicoletti, Adriana G; Cayô, Rodrigo; Gales, Ana C; Alves, Luiz C; Brayner, Fábio B; Vilela, Marinalda A; Morais, Márcia M C

    2015-12-01

    Few reports described the presence of bla(KPC) and qnr genes in the same isolate. This study reports the combination of bla(KPC-2) and qnrB19 genes in Klebsiella pneumoniae ST340 isolate in Brazil. These findings draw attention to this combination in ST340 isolate, which is part of the CC258, disseminated in Latin America.

  9. Complete Genome Sequence of KPC-3- and CTX-M-15-Producing Klebsiella pneumoniae Sequence Type 307

    PubMed Central

    Villa, Laura; Feudi, Claudia; Fortini, Daniela; Iacono, Michele; Bonura, Celestino; Endimiani, Andrea; Mammina, Caterina

    2016-01-01

    Klebsiella pneumoniae sequence type (ST) 307, carrying blaKPC-3, blaCTX-M-15, blaOXA-1, aac(6′)-Ib-cr, and qnrB1 genes, is replacing the predominant hyperepidemic ST258 clone in Italy. Whole-genome and complete plasmid sequencing of one ST307 strain was performed and new features were identified. PMID:27056222

  10. Complete Genome Sequence of Klebsiella pneumoniae Strain HKUOPLC, a Cellulose-Degrading Bacterium Isolated from Giant Panda Feces

    PubMed Central

    Lu, Matthew Guan-Xi; Jiang, Jingwei; Liu, Lirui; Ma, Angel Po-Yee

    2015-01-01

    We report here the complete genome sequence of Klebsiella pneumoniae strain HKUOPLC, isolated from a giant panda fecal sample collected from Ocean Park, Hong Kong. The complete genome of this bacterium may contribute to the discovery of efficient cellulose-degrading pathways. PMID:26564041

  11. Complete Genome Sequence of Klebsiella pneumoniae Strain HKUOPLC, a Cellulose-Degrading Bacterium Isolated from Giant Panda Feces.

    PubMed

    Lu, Matthew Guan-Xi; Jiang, Jingwei; Liu, Lirui; Ma, Angel Po-Yee; Leung, Frederick Chi-Ching

    2015-11-12

    We report here the complete genome sequence of Klebsiella pneumoniae strain HKUOPLC, isolated from a giant panda fecal sample collected from Ocean Park, Hong Kong. The complete genome of this bacterium may contribute to the discovery of efficient cellulose-degrading pathways. Copyright © 2015 Lu et al.

  12. Multiplex PCR for Detection of Seven Virulence Factors and K1/K2 Capsular Serotypes of Klebsiella pneumoniae

    PubMed Central

    Babosan, Ana; Brisse, Sylvain; Genel, Nathalie; Audo, Jennifer; Ailloud, Florent; Kassis-Chikhani, Najiby; Arlet, Guillaume; Decré, Dominique

    2014-01-01

    A single multiplex PCR assay targeting seven virulence factors and the wzi gene specific for the K1 and K2 capsular serotypes of Klebsiella pneumoniae was developed and tested on 65 clinical isolates, which included 45 isolates responsible for community-acquired severe human infections. The assay is useful for the surveillance of emerging highly virulent strains. PMID:25275000

  13. Emerging K1 serotype Klebsiella pneumoniae primary liver abscess: three cases presenting to a single university hospital in Norway

    PubMed Central

    Holmås, Kristoffer; Fostervold, Aasmund; Stahlhut, Steen Gustav; Struve, Carsten; Holter, Jan Cato

    2014-01-01

    Key Clinical Message Community-acquired Klebsiella pneumoniae primary liver abscess (KLA) has been emerging worldwide over the past two decades and with high incidence in Asia. The presence of specific virulence characteristics is a risk factor for a syndrome with metastatic complications. This report signals an increasing emergence in Northern Europe. PMID:25356268

  14. Survey and rapid detection of Klebsiella pneumoniae in clinical samples targeting the rcsA gene in Beijing, China.

    PubMed

    Dong, Derong; Liu, Wei; Li, Huan; Wang, Yufei; Li, Xinran; Zou, Dayang; Yang, Zhan; Huang, Simo; Zhou, Dongsheng; Huang, Liuyu; Yuan, Jing

    2015-01-01

    Klebsiella pneumoniae is a wide-spread nosocomial pathogen. A rapid and sensitive molecular method for the detection of K. pneumoniae in clinical samples is needed to guide therapeutic treatment. In this study, we first described a loop-mediated isothermal amplification (LAMP) method for the rapid detection of capsular polysaccharide synthesis regulating gene rcsA from K. pneumoniaein clinical samples by using two methods including real-time turbidity monitoring and fluorescence detection to assess the reaction. Then dissemination of K. pneumoniae strains was investigated from ICU patients in three top hospitals in Beijing, China. The results showed that the detection limit of the LAMP method was 0.115 pg/μl DNA within 60 min under isothermal conditions (61°C), a 100-fold increase in sensitivity compared with conventional PCR. All 30 non- K. pneumoniae strains tested were negative for LAMP detection, indicating the high specificity of the LAMP reaction. To evaluate the application of the LAMP assay to clinical diagnosis, of 110 clinical sputum samples collected from ICU patients with clinically suspected multi-resistant infections in China, a total of 32 K. pneumoniae isolates were identified for LAMP-based surveillance of rcsA. All isolates belonged to nine different K. pneumoniae multilocus sequence typing (MLST) groups. Strikingly, of the 32 K. pneumoniae strains, 18 contained the Klebsiella pneumoniae Carbapenemase (KPC)-encoding gene bla KPC-2 and had high resistance to β-lactam antibiotics. Moreover, K. pneumoniae WJ-64 was discovered to contain bla KPC-2 and bla NDM-1genes simultaneously in the isolate. Our data showed the high prevalence of bla KPC-2 among K. pneumoniae and co-occurrence of many resistant genes in the clinical strains signal a rapid and continuing evolution of K. pneumoniae. In conclusion, we have developed a rapid and sensitive visual K. pneumoniae detection LAMP assay, which could be a useful tool for clinical screening, on

  15. Hypervirulent Klebsiella pneumoniae induced ventilator-associated pneumonia in mechanically ventilated patients in China.

    PubMed

    Yan, Q; Zhou, M; Zou, M; Liu, W-e

    2016-03-01

    The purpose of this study was to investigate the clinical characteristics of hypervirulent K. pneumoniae (hvKP) induced ventilator-associated pneumonia (VAP) and the microbiological characteristics and epidemiology of the hvKP strains. A retrospective study of 49 mechanically ventilated patients with K. pneumoniae induced VAP was conducted at a university hospital in China from January 2014 to December 2014. Clinical characteristics and K. pneumoniae antimicrobial susceptibility and biofilm formation were analyzed. Genes of capsular serotypes K1, K2, K5, K20, K54 and K57 and virulence factors plasmid rmpA(p-rmpA), iroB, iucA, mrkD, entB, iutA, ybtS, kfu and allS were also evaluated. Multilocus sequence typing (MLST) and random amplified polymorphic DNA (RAPD) analyses were used to study the clonal relationship of the K. pneumoniae strains. Strains possessed p-rmpA and iroB and iucA were defined as hvKP. Of 49 patients, 14 patients (28.6 %) were infected by hvKP. Antimicrobial resistant rate was significantly higher in cKP than that in hvKP. One ST29 K54 extended-spectrum-beta-lactamase (ESBL) producing hvKP strain was detected. The prevalence of K1 and K2 in hvKP was 42.9 % and 21.4 %, respectively. The incidences of K1, K2, K20, p-rmpA, iroB, iucA, iutA, Kfu and alls were significantly higher in hvKP than those in cKP. ST23 was dominant among hvKP strains, and all the ST23 strains had identical RAPD pattern. hvKP has become a common pathogen of VAP in mechanically ventilated patients in China. Clinicians should increase awareness of hvKP induced VAP and enhance epidemiologic surveillance.

  16. In Vitro Pharmacodynamics of Various Antibiotics in Combination against Extensively Drug-Resistant Klebsiella pneumoniae

    PubMed Central

    Lim, Tze-Peng; Cai, Yiying; Hong, Yanjun; Chan, Eric Chun Yong; Suranthran, Sasikala; Teo, Jocelyn Qi-Min; Lee, Winnie Huiling; Tan, Thean-Yen; Hsu, Li-Yang; Koh, Tse-Hsien; Tan, Thuan-Tong

    2015-01-01

    Extensively drug-resistant (XDR) Klebsiella pneumoniae is an emerging pathogen in Singapore. With limited therapeutic options available, combination antibiotics may be the only viable option. In this study, we aimed to elucidate effective antibiotic combinations against XDR K. pneumoniae isolates. Six NDM-1-producing and two OXA-181-producing K. pneumoniae strains were exposed to 12 antibiotics alone and in combination via time-kill studies. A hollow-fiber infection model (HFIM) with pharmacokinetic validation was used to simulate clinically relevant tigecycline-plus-meropenem dosing regimens against 2 XDR K. pneumoniae isolates over 240 h. The emergence of resistance against tigecycline was quantified using drug-free and selective (tigecycline at 3× the MIC) media. The in vitro growth rates were determined and serial passages on drug-free and selective media were carried out on resistant isolates obtained at 240 h. Both the polymyxin B and tigecycline MICs ranged from 1 to 4 mg/liter. In single time-kill studies, all antibiotics alone demonstrated regrowth at 24 h, except for polymyxin B against 2 isolates. Tigecycline plus meropenem was found to be bactericidal in 50% of the isolates. For the isolates that produced OXA-181-like carbapenemases, none of the 55 tested antibiotic combinations was bactericidal. Against 2 isolates in the HFIM, tigecycline plus meropenem achieved a >90% reduction in bacterial burden for 96 h before regrowth was observed until 109 CFU/ml at 240 h. Phenotypically stable and resistant isolates, which were recovered from tigecycline-supplemented plates post-HFIM studies, had lower growth rates than those of their respective parent isolates, possibly implying a substantial biofitness deficit in this population. We found that tigecycline plus meropenem may be a potential antibiotic combination for XDR K. pneumoniae infections, but its efficacy was strain specific. PMID:25691628

  17. Aeromonas punctata derived depolymerase improves susceptibility of Klebsiella pneumoniae biofilm to gentamicin.

    PubMed

    Bansal, Shruti; Harjai, Kusum; Chhibber, Sanjay

    2015-06-11

    To overcome antibiotic resistance in biofilms, enzymes aimed at biofilm dispersal are under investigation. In the present study, applicability of an Aeromonas punctata derived depolymerase capable of degrading the capsular polysaccharide (CPS) of Klebsiella pneumoniae, in disrupting its biofilm and increasing gentamicin efficacy against biofilm was investigated. Intact biofilm of K. pneumoniae was recalcitrant to gentamicin due to lack of antibiotic penetration. On the other hand, gentamicin could not act on disrupted biofilm cells due to their presence in clusters. However, when depolymerase (20 units/ml) was used in combination with gentamicin (10 μg/ml), dispersal of CPS matrix by enzyme facilitated gentamicin penetration across biofilm. This resulted in significant reduction (p < 0.05) in bacterial count in intact and disrupted biofilms. Reduction in CPS after treatment with depolymerase was confirmed by confocal microscopy and enzyme linked lectinosorbent assay. Furthermore, to substantiate our study, the efficacy of bacterial depolymerase was compared with a phage borne depolymerase possessing similar application against K. pneumoniae. Although both were used at same concentration i.e. 20 units/ml, but a higher efficacy of bacterial depolymerase particularly against older biofilms was visibly clear over its phage counterpart. This could be explained due to high substrate affinity (indicated by Km value) and high turnover number (indicated by Kcat value) of the bacterial depolymerase (Km = 89.88 μM, Kcat = 285 s(-1)) over the phage derived one (Km = 150 μM, Kcat = 107 s(-1)). Overall the study indicated that, the A. punctata derived depolymerase possesses antibiofilm potential and improves gentamicin efficacy against K. pneumoniae. Moreover, it can serve as a potential substitute to phage borne depolymerases for treating biofilms formed by K. pneumoniae.

  18. In vitro pharmacodynamics of various antibiotics in combination against extensively drug-resistant Klebsiella pneumoniae.

    PubMed

    Lim, Tze-Peng; Cai, Yiying; Hong, Yanjun; Chan, Eric Chun Yong; Suranthran, Sasikala; Teo, Jocelyn Qi-Min; Lee, Winnie Huiling; Tan, Thean-Yen; Hsu, Li-Yang; Koh, Tse-Hsien; Tan, Thuan-Tong; Kwa, Andrea Lay-Hoon

    2015-05-01

    Extensively drug-resistant (XDR) Klebsiella pneumoniae is an emerging pathogen in Singapore. With limited therapeutic options available, combination antibiotics may be the only viable option. In this study, we aimed to elucidate effective antibiotic combinations against XDR K. pneumoniae isolates. Six NDM-1-producing and two OXA-181-producing K. pneumoniae strains were exposed to 12 antibiotics alone and in combination via time-kill studies. A hollow-fiber infection model (HFIM) with pharmacokinetic validation was used to simulate clinically relevant tigecycline-plus-meropenem dosing regimens against 2 XDR K. pneumoniae isolates over 240 h. The emergence of resistance against tigecycline was quantified using drug-free and selective (tigecycline at 3× the MIC) media. The in vitro growth rates were determined and serial passages on drug-free and selective media were carried out on resistant isolates obtained at 240 h. Both the polymyxin B and tigecycline MICs ranged from 1 to 4 mg/liter. In single time-kill studies, all antibiotics alone demonstrated regrowth at 24 h, except for polymyxin B against 2 isolates. Tigecycline plus meropenem was found to be bactericidal in 50% of the isolates. For the isolates that produced OXA-181-like carbapenemases, none of the 55 tested antibiotic combinations was bactericidal. Against 2 isolates in the HFIM, tigecycline plus meropenem achieved a >90% reduction in bacterial burden for 96 h before regrowth was observed until 10(9) CFU/ml at 240 h. Phenotypically stable and resistant isolates, which were recovered from tigecycline-supplemented plates post-HFIM studies, had lower growth rates than those of their respective parent isolates, possibly implying a substantial biofitness deficit in this population. We found that tigecycline plus meropenem may be a potential antibiotic combination for XDR K. pneumoniae infections, but its efficacy was strain specific. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  19. Increasing incidence of carbapenemase-producing Escherichia coli and Klebsiella pneumoniae in Belgian hospitals.

    PubMed

    De Laveleye, M; Huang, T D; Bogaerts, P; Berhin, C; Bauraing, C; Sacré, P; Noel, A; Glupczynski, Y

    2017-01-01

    Carbapenemase-producing Enterobacteriaceae are increasingly reported worldwide. The aim of the study was to determine the incidence and molecular epidemiology of carbapenemase-producing (CP) Escherichia coli and Klebsiella pneumoniae (CP-E/K) in Belgium. Eleven hospital-based laboratories collected carbapenem non-susceptible (CNS) isolates of E. coli and K. pneumoniae detected in clinical specimens from January 2013 to December 2014. All CNS strains were tested for carbapenemase production and typed by multilocus sequence typing (MLST) for a 6-month period as part of the European Survey on Carbapenemase-Producing Enterobacteriaceae in Europe (EuSCAPE) structured survey. In addition, an equal number of carbapenem-susceptible isolates collected were preserved as a control group for risk factor analysis. The overall incidence rate of CP-E/K isolates in hospitals increased from 0.124 in 2013 to 0.223 per 1000 admissions in 2014. From November 2013 to April 2014, 30 CP K. pneumoniae [OXA-48 (n = 16), KPC (n = 13), OXA-427 (n = 1)] and five CP E. coli [OXA-48 (n = 3), NDM (n = 1), OXA-427 (n = 1)] isolates were detected in ten hospitals. The 16 OXA-48-producing K. pneumoniae strains were distributed into eight sequence types (STs), while the 13 KPC-producing K. pneumoniae clustered into three STs dominated by ST512 (n = 7) and ST101 (n = 5). Compared to controls, we observed among CP-E/K carriers significantly higher proportion of males, respiratory origins, previous hospitalization, nosocomial setting, and a significantly lower proportion of bloodstream infections. Our study confirms the rapid spread of CP-E/K in Belgian hospitals and the urgent need for a well-structured and coordinated national surveillance plan in order to limit their dissemination.

  20. Asian sand dust enhances murine lung inflammation caused by Klebsiella pneumoniae

    SciTech Connect

    He, Miao; Ichinose, Takamichi; Yoshida, Seiichi; Yamamoto, Shoji; Inoue, Ken-ichiro; Takano, Hirohisa; Yanagisawa, Rie; Nishikawa, Masataka; Mori, Ikuko; Sun, Guifan; Shibamoto, Takayuki

    2012-01-15

    Inhaling concomitants from Asian sand dust (ASD) may result in exacerbation of pneumonia by the pathogen. The exacerbating effect of ASD on pneumonia induced by Klebsiella pneumoniae (KP) was investigated in ICR mice. The organic substances adsorbed onto ASD collected from the atmosphere of Iki-island in Japan were excluded by heat treatment at 360 °C for 30 min. ICR mice were instilled intratracheally with ASD at doses of 0.05 mg or 0.2 mg/mouse four times at 2-week intervals (total dose of 0.2 mg or 0.8 mg/mouse) and were administrated with ASD in the presence or absence of KP at the last intratracheal instillation. Pathologically, ASD caused exacerbation of pneumonia by KP as shown by increased inflammatory cells within the bronchiolar and the alveolar compartments. ASD enhanced the neutrophil number dose dependently as well as the expression of cytokines (IL-1β, IL-6, IL-12, IFN-γ, TNF-α) and chemokines (KC, MCP-1, MIP-1α) related to KP in BALF. In an in vitro study using RAW264.7 cells, combined treatment of ASD and KP increased gene expression of IL-1β, IL-6, IFN-β, KC, MCP-1, and MIP-1α. The same treatment tended to increase the protein level of IL-1β, TNF-α and MCP-1 in a culture medium compared to each treatment alone. The combined treatment tended to increase the gene expression of Toll-like receptor 2 (TLR2), and NALP3, ASC and caspase-1 compared with KP alone. These results suggest that the exacerbation of pneumonia by ASD + KP was due to the enhanced production of pro-inflammatory mediators via activation of TLR2 and NALP3 inflammasome pathways in alveolar macrophages.

  1. Physiological and molecular characteristics of carbapenem resistance in Klebsiella pneumoniae and Enterobacter aerogenes.

    PubMed

    Pereira, Rito Santo; Dias, Vanessa Cordeiro; Ferreira-Machado, Alessandra Barbosa; Resende, Juliana Alves; Bastos, André Netto; Andrade Bastos, Lucas Quinet; Andrade Bastos, Victor Quinet; Bastos, Ricardo Villela; Da Silva, Vânia Lúcia; Diniz, Cláudio Galuppo

    2016-06-30

    Bacterial resistance is a growing concern in the nosocomial environment in which Klebsiella pneumoniae and Enterobacter aerogenes play an important role due to their opportunism and carbapenemase-production. This work aimed to evaluate physiological and molecular characteristics of carbapenem-resistant K. pneumoniae and E. aerogenes isolated in a Brazilian tertiary hospital. In total, 42 carbapenem-resistant bacteria isolated from clinical specimens were included (21 K. pneumoniae and 21 E. aerogenes). Drug-sensitive K. pneumoniae (n = 27) were also included. Antimicrobial susceptibility and biocide tolerance patterns, hemolytic activity, tolerance to oxidative stress, and aggregative ability were assessed. Genetic markers related to carbapenem resistance, or ESBL-production were screened by PCR. Compared to drug-sensitive strains, carbapenem-resistant K. pneumoniae were more tolerant to biocides and to oxidative stress, and they displayed an increase in biofilm formation. The genetic markers blaKPC (95.2%) and blaTEM (90.5%) were the most frequent. Among the carbapenem-resistant E. aerogenes strains, blaKPC, and blaTEM were detected in all bacteria. Drug-sensitive E. aerogenes were not isolated in the same period. blaSHV, blaVIM, and blaCTX markers were also observed among carbapenem-resistant bacteria. Results suggest that carbapenemase-producing enterobacteria might show peculiar characteristics regarding their physiology associated with their environmental persistency, virulence, and multidrug resistance. The observed phenomenon may have implications not only for antimicrobial chemotherapy, but also for the prognosis of infectious diseases and infection control.

  2. Efflux Pump, the Masked Side of ß-Lactam Resistance in Klebsiella pneumoniae Clinical Isolates

    PubMed Central

    Pages, Jean-Marie; Lavigne, Jean-Philippe; Leflon-Guibout, Véronique; Marcon, Estelle; Bert, Frédéric; Noussair, Latifa; Nicolas-Chanoine, Marie-Hélène

    2009-01-01

    Background β-lactamase production and porin decrease are the well-recognized mechanisms of acquired ß-lactam resistance in Klebsiella pneumoniae isolates. However, such mechanisms proved to be absent in K. pneumoniae isolates that are non susceptible to cefoxitin (FOX) and succeptible to amoxicillin+clavulanic acid in our hospital. Assessing the role of efflux pumps in this β-lactam phenotype was the aim of this study. Methodology/Findings MICs of 9 β-lactams, including cloxacillin (CLX), and other antibiotic families were tested alone and with an efflux pump inhibitor (EPI), then with both CLX (subinhibitory concentrations) and EPI against 11 unique bacteremia K. pneumoniae isolates displaying the unusual phenotype, and 2 ATCC strains. CLX and EPI-dose dependent effects were studied on 4 representatives strains. CLX MICs significantly decreased when tested with EPI. A similar phenomenon was observed with piperacillin+tazobactam whereas MICs of the other β-lactams significantly decreased only in the presence of both EPI and CLX. Thus, FOX MICs decreased 128 fold in the K. pneumoniae isolates but also16 fold in ATCC strain. Restoration of FOX activity was CLX dose-dependent suggesting a competitive relationship between CLX and the other β-lactams with regard to their efflux. For chloramphenicol, erythromycin and nalidixic acid whose resistance was also due to efflux, adding CLX to EPI did not increase their activity suggesting differences between the efflux process of these molecules and that of β-lactams. Conclusion This is the first study demonstrating that efflux mechanism plays a key role in the β-lactam susceptibility of clinical isolates of K. pneumoniae. Such data clearly evidence that the involvement of efflux pumps in ß-lactam resistance is specially underestimated in clinical isolates. PMID:19279676

  3. Rise and fall of KPC-producing Klebsiella pneumoniae in New York City.

    PubMed

    Abdallah, Marie; Olafisoye, Olawole; Cortes, Christopher; Urban, Carl; Landman, David; Ghitan, Monica; Collins, BeverlyAnn; Bratu, Simona; Quale, John

    2016-10-01

    The study objective was to examine the epidemiological trends of KPC-producing Klebsiella pneumoniae in New York City medical centres. Single patient isolates of K. pneumoniae were collected from nine medical centres in New York City during a 3 month period from 2013 to 2014. Isolates were tested for the presence of blaKPC. Results were compared with similar surveillance studies conducted in 2006 and 2009. Infection control data, including utilization of medical devices, were analysed at a subset of hospitals. There was a progressive decline in the percentage of K. pneumoniae harbouring blaKPC from 2006 to 2013-14. For the nine hospitals that participated in all three surveillance studies, the percentages of isolates with blaKPC fell from 36% in 2006 to 25% in 2009 to 13% in 2013-14. Seven of the nine hospitals had marked declines in isolates with blaKPC, while two hospitals continued to struggle with this pathogen. These two hospitals were smaller and had longer lengths of patient stay. Device utilization rates were obtained from two hospitals that successfully controlled the spread of KPC-producing K. pneumoniae; both had ∼20%-25% reduction in the usage of urinary catheters. Changes in antibiotic usage at one hospital could not explain the decline in these pathogens. Over the past decade there has been a steady decline in KPC-producing K. pneumoniae in most New York City hospitals. The reason for the decline is probably multifactorial, involving a reduction in device (catheter) utilization and possibly an improvement in infection control practices. © The Author 2016. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  4. Risk factors for outbreaks of multidrug-resistant Klebsiella pneumoniae in critical burn patients.

    PubMed

    Sánchez, Manuel; Herruzo, Rafael; Marbán, Alvaro; Araujo, Pilar; Asensio, Maria J; Leyva, Francisco; Casado, Cesar; García-de-Lorenzo, Abelardo

    2012-01-01

    The objective of this study is to identify the risk factors related to colonization or infection in an outbreak of multidrug-resistant Klebsiella pneumoniae in a burn patient unit. The authors studied the risk factors associated with colonization or infection using a case-control study design involving patients with multidrug resistant K. pneumoniae (n = 26) and controls (n = 50). They describe the outbreak and provide a retrospective analysis that encompasses patient demographics, microbiological isolation, culture sites, burn features, inhalation injury, biomarkers (lactate and N-terminal probrain natriuretic peptide), general illness severity scores (Acute Physiology and Chronic Health Evaluation II and Sepsis-related Organ Failure Assessment), burn-specific severity scores such as the Abbreviated Burn Severity Index (ABSI), length of stay, and mortality. Patients colonized with multidrug-resistant K. pneumoniae were older (55 vs 42 years), presented with larger burns (32 vs 18% of BSA), and more frequently had full-thickness burns (53 vs 22%). They also had higher ABSI, Acute Physiology and Chronic Health Evaluation II, and Sepsis-related Organ Failure Assessment scores, and they required more days of mechanical ventilation and longer stays in the critical burn unit. Multivariate analysis showed that the factors most significantly related to the development of infection or colonization with K. pneumoniae were burns located on head and neck (odds ratio, 4.81) and the ABSI score (odds ratio, 1.66). Control of the outbreak was achieved by enforcing contact precautions and extensive cleaning. An elevated ABSI score and burns located on the head and neck were the risk factors most significantly related to colonization or infection in an outbreak of multidrug-resistant K. pneumoniae in a critical burn patient unit.

  5. Asian sand dust enhances murine lung inflammation caused by Klebsiella pneumoniae.

    PubMed

    He, Miao; Ichinose, Takamichi; Yoshida, Seiichi; Yamamoto, Shoji; Inoue, Ken-ichiro; Takano, Hirohisa; Yanagisawa, Rie; Nishikawa, Masataka; Mori, Ikuko; Sun, Guifan; Shibamoto, Takayuki

    2012-01-15

    Inhaling concomitants from Asian sand dust (ASD) may result in exacerbation of pneumonia by the pathogen. The exacerbating effect of ASD on pneumonia induced by Klebsiella pneumoniae (KP) was investigated in ICR mice. The organic substances adsorbed onto ASD collected from the atmosphere of Iki-island in Japan were excluded by heat treatment at 360°C for 30min. ICR mice were instilled intratracheally with ASD at doses of 0.05mg or 0.2mg/mouse four times at 2-week intervals (total dose of 0.2mg or 0.8mg/mouse) and were administrated with ASD in the presence or absence of KP at the last intratracheal instillation. Pathologically, ASD caused exacerbation of pneumonia by KP as shown by increased inflammatory cells within the bronchiolar and the alveolar compartments. ASD enhanced the neutrophil number dose dependently as well as the expression of cytokines (IL-1β, IL-6, IL-12, IFN-γ, TNF-α) and chemokines (KC, MCP-1, MIP-1α) related to KP in BALF. In an in vitro study using RAW264.7 cells, combined treatment of ASD and KP increased gene expression of IL-1β, IL-6, IFN-β, KC, MCP-1, and MIP-1α. The same treatment tended to increase the protein level of IL-1β, TNF-α and MCP-1 in a culture medium compared to each treatment alone. The combined treatment tended to increase the gene expression of Toll-like receptor 2 (TLR2), and NALP3, ASC and caspase-1 compared with KP alone. These results suggest that the exacerbation of pneumonia by ASD+KP was due to the enhanced production of pro-inflammatory mediators via activation of TLR2 and NALP3 inflammasome pathways in alveolar macrophages.

  6. Functional Genomic Screen Identifies Klebsiella pneumoniae Factors Implicated in Blocking Nuclear Factor κB (NF-κB) Signaling.

    PubMed

    Tomás, Anna; Lery, Leticia; Regueiro, Verónica; Pérez-Gutiérrez, Camino; Martínez, Verónica; Moranta, David; Llobet, Enrique; González-Nicolau, Mar; Insua, Jose L; Tomas, Juan M; Sansonetti, Philippe J; Tournebize, Régis; Bengoechea, José A

    2015-07-03

    Klebsiella pneumoniae is an etiologic agent of community-acquired and nosocomial pneumonia. It has been shown that K. pneumoniae infections are characterized by reduced early inflammatory response. Recently our group has shown that K. pneumoniae dampens the activation of inflammatory responses by antagonizing the activation of the NF-κB canonical pathway. Our results revealed that K. pneumoniae capsule polysaccharide (CPS) was necessary but not sufficient to attenuate inflammation. To identify additional Klebsiella factors required to dampen inflammation, we standardized and applied a high-throughput gain-of-function screen to examine a Klebsiella transposon mutant library. We identified 114 mutants that triggered the activation of NF-κB. Two gene ontology categories accounted for half of the loci identified in the screening: metabolism and transport genes (32% of the mutants) and envelope-related genes (17%). Characterization of the mutants revealed that the lack of the enterobactin siderophore was linked to a reduced CPS expression, which in turn underlined the NF-κB activation induced by the mutant. The lipopolysaccharide (LPS) O-polysaccharide and the pullulanase (PulA) type 2 secretion system (T2SS) are required for full effectiveness of the immune evasion. Importantly, these factors do not play a redundant role. The fact that LPS O-polysaccharide and T2SS mutant-induced responses were dependent on TLR2-TLR4-MyD88 activation suggested that LPS O-polysaccharide and PulA perturbed Toll-like receptor (TLR)-dependent recognition of K. pneumoniae. Finally, we demonstrate that LPS O-polysaccharide and pulA mutants are attenuated in the pneumonia mouse model. We propose that LPS O-polysaccharide and PulA T2SS could be new targets for the design of new antimicrobials. Increasing TLR-governed defense responses might provide also selective alternatives for the management of K. pneumoniae pneumonia.

  7. Functional Genomic Screen Identifies Klebsiella pneumoniae Factors Implicated in Blocking Nuclear Factor κB (NF-κB) Signaling*

    PubMed Central

    Tomás, Anna; Lery, Leticia; Regueiro, Verónica; Pérez-Gutiérrez, Camino; Martínez, Verónica; Moranta, David; Llobet, Enrique; González-Nicolau, Mar; Insua, Jose L.; Tomas, Juan M.; Sansonetti, Philippe J.; Tournebize, Régis; Bengoechea, José A.

    2015-01-01

    Klebsiella pneumoniae is an etiologic agent of community-acquired and nosocomial pneumonia. It has been shown that K. pneumoniae infections are characterized by reduced early inflammatory response. Recently our group has shown that K. pneumoniae dampens the activation of inflammatory responses by antagonizing the activation of the NF-κB canonical pathway. Our results revealed that K. pneumoniae capsule polysaccharide (CPS) was necessary but not sufficient to attenuate inflammation. To identify additional Klebsiella factors required to dampen inflammation, we standardized and applied a high-throughput gain-of-function screen to examine a Klebsiella transposon mutant library. We identified 114 mutants that triggered the activation of NF-κB. Two gene ontology categories accounted for half of the loci identified in the screening: metabolism and transport genes (32% of the mutants) and envelope-related genes (17%). Characterization of the mutants revealed that the lack of the enterobactin siderophore was linked to a reduced CPS expression, which in turn underlined the NF-κB activation induced by the mutant. The lipopolysaccharide (LPS) O-polysaccharide and the pullulanase (PulA) type 2 secretion system (T2SS) are required for full effectiveness of the immune evasion. Importantly, these factors do not play a redundant role. The fact that LPS O-polysaccharide and T2SS mutant-induced responses were dependent on TLR2-TLR4-MyD88 activation suggested that LPS O-polysaccharide and PulA perturbed Toll-like receptor (TLR)-dependent recognition of K. pneumoniae. Finally, we demonstrate that LPS O-polysaccharide and pulA mutants are attenuated in the pneumonia mouse model. We propose that LPS O-polysaccharide and PulA T2SS could be new targets for the design of new antimicrobials. Increasing TLR-governed defense responses might provide also selective alternatives for the management of K. pneumoniae pneumonia. PMID:25971969

  8. Therapeutic effects of cefotiam and cefazolin on experimental pneumonia caused by Klebsiella pneumoniae DT-S in mice.

    PubMed Central

    Nishi, T; Tsuchiya, K

    1980-01-01

    The efficacies of several dosage schedules, productive of plasma levels of cefotiam and cefazolin of short and long duration and starting at three levels of cefotiam and cefazolin of short and long duration and starting at three different times (3, 18, and 30h) after infection, were examined in experimental pneumonia caused by Klebsiella pneumoniae DT-S in mice. With each of the multiday regimens there was a large segment of the day when plasma levels fell below assayable concentrations. In all cases, cefotiam proved about eight times as active as cefazolin, indicating that the potent in vitro antibacterial activity of cefotiam was well reflected in the therapeutic effect in this model infection. As judged by the total dose administered, the regimen of cefotiam producing a low but sustained plasma level gave better therapeutic effects than that exhibiting a high but transient plasma level. The cefotiam levels in the plasma of mice that received the regimen effective when initiated at 18 h after infection were less than the expected levels in humans after intravenous infusion of the usual clinical dose. PMID:7004342

  9. Effect of a Metalloantibiotic Produced by Pseudomonas aeruginosa on Klebsiella pneumoniae Carbapenemase (KPC)-producing K. pneumoniae.

    PubMed

    Kerbauy, Gilselena; Vivan, Ana C P; Simões, Glenda C; Simionato, Ane S; Pelisson, Marsileni; Vespero, Eliana C; Costa, Silvia F; Andrade, Celia G T de J; Barbieri, Daiane M; Mello, João C P; Morey, Alexandre T; Yamauchi, Lucy M; Yamada-Ogatta, Sueli F; de Oliveira, Admilton G; Andrade, Galdino

    2016-01-01

    Multidrug-resistant organisms (MDRO) are a great problem in hospitals, where thousands of people are infected daily, with the occurrence of high mortality rates, especially in infections caused by Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC-producing Kpn). The challenge is to find new compounds that can control KPC producing-Kpn infections. The aim of this study was to evaluate the antibiotic activity of the F3d fraction produced by the Pseudomonas aeruginosa LV strain against clinical isolates of KPC-producing Kpn. The results showed that the minimum inhibitory concentration of F3d (62.5 µg mL(-1)), containing an organic metallic compound, killed planktonic cells of KPC-producing Kpn strains after 30 min of incubation. At the same concentration, this fraction also showed an inhibitory effect against biofilm of these bacteria after 24 h of incubation. Treatment with the F3d fraction caused pronounced morphological alterations in both planktonic and biofilm cells of the bacteria. The inhibitory effect of the F3d fraction seems to be more selective for the bacteria than the host cells, indicating its potential in the development of new drugs for the treatment of infections caused by KPC-producing Kpn and other MDRO.

  10. Klebsiella pneumoniae Capsule Polysaccharide Impedes the Expression of β-Defensins by Airway Epithelial Cells▿

    PubMed Central

    Moranta, David; Regueiro, Verónica; March, Catalina; Llobet, Enrique; Margareto, Javier; Larrate, Eider; Garmendia, Junkal; Bengoechea, José A.

    2010-01-01

    Human β-defensins (hBDs) contribute to the protection of the respiratory tract against pathogens. It is reasonable to postulate that pathogens have developed countermeasures to resist them. Klebsiella pneumoniae capsule polysaccharide (CPS), but not the lipopolysaccharide O antigen, mediated resistance against hBD1 and hBD2. hBD3 was the most potent hBD against Klebsiella. We investigated the possibility that as a strategy for survival in the lung, K. pneumoniae may not activate the expression of hBDs. Infection of A549 and normal human bronchial cells with 52145-ΔwcaK2, a CPS mutant, increased the expression of hBD2 and hBD3. Neither the wild type nor the lipopolysaccharide O antigen mutant increased the expression of hBDs. In vivo, 52145-ΔwcaK2 induced higher levels of mBD4 and mBD14, possible mouse orthologues of hBD2 and hBD3, respectively, than the wild type. 52145-ΔwcaK2-dependent upregulation of hBD2 occurred via NF-κB and mitogen-activated protein kinases (MAPKs) p44/42, Jun N-terminal protein kinase (JNK)-dependent pathways. The increase in hBD3 expression was dependent on the MAPK JNK. 52145-ΔwcaK2 engaged Toll-like receptors 2 and 4 (TLR2 and TLR4) to activate hBD2, whereas hBD3 expression was dependent on NOD1. K. pneumoniae induced the expression of CYLD and MKP-1, which act as negative regulators for 52145-ΔwcaK2-induced expression of hBDs. Bacterial engagement of pattern recognition receptors induced CYLD and MKP-1, which may initiate the attenuation of proinflammatory pathways. The results of this study indicate that K. pneumoniae CPS not only protects the pathogen from the bactericidal action of defensins but also impedes their expression. These features of K. pneumoniae CPS may facilitate pathogen survival in the hostile environment of the lung. PMID:20008534

  11. Generation and Validation of the iKp1289 Metabolic Model for Klebsiella pneumoniae KPPR1.

    PubMed

    Henry, Christopher S; Rotman, Ella; Lathem, Wyndham W; Tyo, Keith E J; Hauser, Alan R; Mandel, Mark J

    2017-02-15

    Klebsiella pneumoniae has a reputation for causing a wide range of infectious conditions, with numerous highly virulent and antibiotic-resistant strains. Metabolic models have the potential to provide insights into the growth behavior, nutrient requirements, essential genes, and candidate drug targets in these strains. Here we develop a metabolic model for KPPR1, a highly virulent strain of K. pneumoniae. We apply a combination of Biolog phenotype data and fitness data to validate and refine our KPPR1 model. The final model displays a predictive accuracy of 75% in identifying potential carbon and nitrogen sources for K. pneumoniae and of 99% in predicting nonessential genes in rich media. We demonstrate how this model is useful in studying the differences in the metabolic capabilities of the low-virulence MGH 78578 strain and the highly virulent KPPR1 strain. For example, we demonstrate that these strains differ in carbohydrate metabolism, including the ability to metabolize dulcitol as a primary carbon source. Our model makes numerous other predictions for follow-up verification and analysis. Published by Oxford University Press for the Infectious Diseases Society of America 2017. This work is written by (a) US Government employee(s) and is in the public domain in the US.

  12. Infections with blaKPC-2-producing Klebsiella pneumoniae in renal transplant patients: a retrospective study.

    PubMed

    Cicora, F; Mos, F; Paz, M; Allende, N G; Roberti, J

    2013-11-01

    In renal transplant recipients, the urinary tract is the most common site of infections that might be caused by pathogens while on immunosuppressive therapy. The spread of enterobacteria resistant to carbapenem is worrying, as it is generally used as this agent is the first-line therapy for infections caused by Enterobacteriaceae producing extended spectrum β-lactamases. The most frequently encountered class A carbapenemases are the Klebsiella pneumoniae carbapenemase (KPC) enzymes. We describe the treatment and outcomes of 6 renal transplant patients who had urinary tract infections (UTIs) with blaKPC-2-producing K pneumoniae, confirmed by polymerase chain reaction amplification, namely 13.33% of renal transplant patients in the study period. Four patients survived, including 1 with reinfections and relapse, and 2 patients died. The antibiotics used for treatment, alone or combined, were colistin (n = 6, 42.8%), tigecycline (n = 5, 35.7%), doxycycline (n = 3, 21.4%), meropenem (n = 3, 21.4%), and fosfomycyn (n = 1, 7%). UTIs caused by carbapenemase-producing K pneumoniae are life-threatening. In the cases presented, favorable results were achieved with monotherapies using colistin, doxycycline, or meropenem.

  13. Genomic and transcriptomic analysis of NDM-1 Klebsiella pneumoniae in spaceflight reveal mechanisms underlying environmental adaptability

    PubMed Central

    Li, Jia; Liu, Fei; Wang, Qi; Ge, Pupu; Woo, Patrick C. Y.; Yan, Jinghua; Zhao, Yanlin; Gao, George F.; Liu, Cui Hua; Liu, Changting

    2014-01-01

    The emergence and rapid spread of New Delhi Metallo-beta-lactamase-1 (NDM-1)-producing Klebsiella pneumoniae strains has caused a great concern worldwide. To better understand the mechanisms underlying environmental adaptation of those highly drug-resistant K. pneumoniae strains, we took advantage of the China's Shenzhou 10 spacecraft mission to conduct comparative genomic and transcriptomic analysis of a NDM-1 K. pneumoniae strain (ATCC BAA-2146) being cultivated under different conditions. The samples were recovered from semisolid medium placed on the ground (D strain), in simulated space condition (M strain), or in Shenzhou 10 spacecraft (T strain) for analysis. Our data revealed multiple variations underlying pathogen adaptation into different environments in terms of changes in morphology, H2O2 tolerance and biofilm formation ability, genomic stability and regulation of metabolic pathways. Additionally, we found a few non-coding RNAs to be differentially regulated. The results are helpful for better understanding the adaptive mechanisms of drug-resistant bacterial pathogens. PMID:25163721

  14. Characteristics of community-onset NDM-1-producing Klebsiella pneumoniae isolates.

    PubMed

    Kim, So Yeon; Rhee, Ji-Young; Shin, Sang Yop; Ko, Kwan Soo

    2014-01-01

    Multilocus sequence typing and in vitro antimicrobial susceptibility testing were performed for three community-onset New Delhi metallo-β-lactamase-1 (NDM-1)-producing Klebsiella pneumoniae isolates from Korea. The genetic structure surrounding the blaNDM-1 gene was determined in blaNDM-1-harbouring plasmids. Three NDM-1-producing K. pneumoniae isolates were found to belong to the same clone (sequence type 340). Each of these isolates showed the same genetic structure surrounding the blaNDM-1 gene. The genes blaNDM-1, bleMBL, trpF and dsbC were flanked by two intact insertion sequences, ISAba125 and IS26, which may promote horizontal gene transfer. The blaNDM-1-harbouring plasmids conferred antimicrobial resistance to carbapenems, cephalosporins, aminoglycosides and aztreonam in transconjugants. It can be speculated that either the entire blaNDM-1-harbouring plasmids or just the part of the plasmid containing the blaNDM-1 gene may have transferred between K. pneumoniae and Escherichia coli. Following the transfer, the isolate disseminated throughout Korea. This study suggests the need for monitoring the dissemination of NDM-1-producing isolates across countries or continents due to their potential transferability via ISAba125- and IS26-associated transposons.

  15. Development of an optimized random amplified polymorphic DNA protocol for fingerprinting of Klebsiella pneumoniae.

    PubMed

    Ashayeri-Panah, M; Eftekhar, F; Feizabadi, M M

    2012-04-01

    To develop an optimized random amplified polymorphic DNA (RAPD) protocol for fingerprinting clinical isolates of Klebsiella pneumoniae. Employing factorial design of experiments, repeatable amplification patterns were obtained for 54 nosocomial isolates using 1 μmol 1(-1) primer, 4 mmol 1(-1) MgCl(2), 0·4 mmol 1(-1) dNTPs, 2·5 U Taq DNA polymerase and 90 ng DNA template in a total volume of 25 μl. The optimum thermocycling program was: initial denaturation at 94°C for 4 min followed by 50 cycles of 1 min at 94°C, 2 min at 34°C, 2 min at 72°C and a final extension at 72°C for 10 min. The optimized RAPD protocol was highly discriminatory (Simpson's diversity index, 0·982), and all isolates were typable with repeatable patterns (Pearson's similarity coefficient ≈ 100%). Seven main clusters were obtained on a similarity level of 70% and 32 distinct clusters on a similarity level of 85%, reflecting the heterogeneity of the isolates. Systematic optimization of RAPD generated reliable DNA fingerprints for nosocomial isolates of K. pneumoniae. This is the first report on RAPD optimization based on factorial design of experiments for discrimination of K. pneumoniae. © 2012 The Authors. Letters in Applied Microbiology © 2012 The Society for Applied Microbiology.

  16. Colistin resistance associated with outer membrane protein change in Klebsiella pneumoniae and Enterobacter asburiae.

    PubMed

    Kádár, Béla; Kocsis, Béla; Tóth, Ákos; Kristóf, Katalin; Felső, Péter; Kocsis, Béla; Böddi, Katalin; Szabó, Dóra

    2017-06-01

    In this study, outer membrane proteins (OMPs) of colistin-resistant Klebsiella pneumoniae and Enterobacter asburiae were analyzed. One colistin-susceptible and three colistin-resistant K. pneumoniae sequence type 258 strains as well as one colistin-susceptible E. asburiae and its colistin-heteroresistant counterpart strain were involved in the study. OMP analysis of each strain was performed by microchip method. Matrix-assisted laser desorption ionization time of flight/mass spectrometry (MALDI-TOF/MS) investigation was carried out after separation of OMPs by two-dimensional gel electrophoresis and in-gel digestion. The MALDI-TOF/MS analysis of OMPs in the colistin-susceptible K. pneumoniae found 16 kDa proteins belonging to the LysM domain/BON superfamily, as well as DNA starvation proteins, whereas OmpX and OmpW were detected in the colistin-resistant counterpart strains. OmpC and OmpW were detected in the colistin-susceptible E. asburiae, whereas OmpA and OmpX were identified in the colistin-resistant counterpart. This study demonstrated that OMP differences were between colistin-susceptible and -resistant counterpart strains. The altered Gram-negative cell wall may contribute to acquired colistin resistance in Enterobacteriaceae.

  17. Consequences of cps mutation of Klebsiella pneumoniae on 1,3-propanediol fermentation.

    PubMed

    Guo, Ni-Ni; Zheng, Zong-Ming; Mai, Yu-Lin; Liu, Hong-Juan; Liu, De-Hua

    2010-03-01

    The filtration in 1,3-propanediol (1,3-PD) downstream process is influenced by the large amounts of capsular polysaccharides (CPS) produced by Klebsiella pneumoniae CGMCC 1.6366. The morphological and fermentation properties were investigated with the CPS-deficient mutant K. pneumoniae CGMCC 1.6366 CPS. Similar biomass was obtained with CGMCC 1.6366, and the mutant strain in batch cultures indicating the cell growth was slightly inhibited by CPS defection. The viscosity of fermentation broth by mutant strain decreased by 27.45%. The flux with ceramic membrane filter was enhanced from 168.12 to 303.6 l h(-1) m(-2), exhibiting the great importance for downstream processing of 1,3-PD fermentation. The products spectrum of mutant isolate changed remarkably regarding to the concentration of fermentation products. The synthesis of important 1,3-PD and 2,3-butanediol was enhanced from 9.73 and 4.06 g l(-1) to 10.37 and 4.77 g l(-1) in batch cultures. The noncapsuled K. pneumoniae provided higher 1,3-PD yield of 0.54 mol mol(-1) than that of encapsuled wild parent in batch cultures. The fed-batch fermentation of mutant strain resulted in 1,3-PD concentration, yield, and productivity of 78.13 g l(-1), 0.53 mol mol(-1), and 1.95 g l(-1) h(-1), respectively.

  18. Nosocomial infections by Klebsiella pneumoniae carbapenemase producing enterobacteria in a teaching hospital

    PubMed Central

    Seibert, Gabriela; Hörner, Rosmari; Meneghetti, Bettina Holzschuh; Righi, Roselene Alves; Forno, Nara Lucia Frasson Dal; Salla, Adenilde

    2014-01-01

    Objective To analyze the profile of patients with microorganisms resistant to carbapenems, and the prevalence of the enzyme Klebsiella pneumoniae carbapenemase in interobacteriaceae. Methods Retrospective descriptive study. From the isolation in bacteriological tests ordered by clinicians, we described the clinical and epidemiological characteristics of patients with enterobacteria resistants to carbapenems at a university hospital, between March and October 2013. Results We included 47 isolated patients in this study, all exhibiting resistance to carbapenems, including 9 patients who were confirmed as infected/colonized with K. pneumoniae carbapenemase. Isolation in tracheal aspirates (12; 25.5%) predominated. The resistance to ertapenem, meropenem, and imipenem was 91.5%, 83.0% and 80.0%, respectively. Aminoglycosides was the class of antimicrobials that showed the highest sensitivity, 91.5% being sensitive to amikacin and 57.4% to gentamicin. Conclusion The K. pneumoniae carbapenemase was an important agent in graun isotaling in hospital intection. The limited therapeutic options emphasize the need for rapid laboratory detection, as well as the implementation of measures to prevent and control the spread of these pathogens. PMID:25295446

  19. Integron mediated multidrug resistance in extended spectrum beta-lactamase producing clinical isolates of Klebsiella pneumoniae

    PubMed Central

    Mobarak-Qamsari, Maryam; Ashayeri-Panah, Mitra; Eftekhar, Freshteh; Feizabadi, Mohammad Mehdi

    2013-01-01

    The present study describes integron mediated multiple antibiotic resistance in extended-spectrum β-lactamase producing clinical isolates of Klebsiella pneumoniae. One hundred and four clinical isolates of K. pneumoniae from two Iranian hospitals were screened for extended-spectrum β-lactamase production and susceptibility of the extended-spectrum β-lactamase producing isolates was determined to 17 antibiotics by disc diffusion. Presence of integron classes 1, 2 and 3 was detected by PCR and integrase specific primers. Isolates harboring class 1 integron were then screened for variable regions using PCR. Fifty isolates (48%) produced extended-spectrum β-lactamases among which, 22 (44%) harbored class 1, 3 (6%) carried class 2 and none contained class 3 integons. Integron carriage was significantly associated with higher rates of multiple antibiotic resistance in extended-spectrum β-lactamase producing clinical isolates of K. pneumoniae. Integron harboring isolates were more resistant to aztreonam (51.3%), ceftazidime (42.6%), cefotaxime (43.3%), cefepime (24.6%), kanamycin (43.2%), tobramycin (30.7%), norfloxcacin (32%) and spectinomycin (25.6%) compared to the organisms without integrons. On the other hand, resistance to nitrofurantoin and streptomycin was significantly higher among the integron negative isolates. PCR amplification of class1 integron variable regions revealed 9 different sized DNA fragments and isolates with similar profiles for class 1 integron variable regions showed the same antibiotic resistance phenotypes. PMID:24516451

  20. Multicellularity and Antibiotic Resistance in Klebsiella pneumoniae Grown Under Bloodstream-Mimicking Fluid Dynamic Conditions

    PubMed Central

    Thornton, Margaret M.; Chung-Esaki, Hangyul M.; Irvin, Charlene B.; Bortz, David M.; Solomon, Michael J.; Younger, John G.

    2012-01-01

    Background. While the importance of fluid dynamical conditions is well recognized in the growth of biofilms, their role during bacteremia is unknown. We examined the impact of physiological fluid shear forces on the development of multicellular aggregates of Klebsiella pneumoniae. Methods. Wild-type and O-antigen or capsular mutants of K. pneumoniae were grown as broth culture in a Taylor-Couette flow cell configured to provide continuous shear forces comparable to those encountered in the human arterial circulation (ie, on the order of 1.0 Pa). The size distribution and antibiotic resistance of aggregates formed in this apparatus were determined, as was their ability to persist in the bloodstream of mice following intravenous injection. Results. Unlike growth in shaking flasks, bacteria grown in the test apparatus readily formed aggregates, a phenotype largely absent in capsular mutants and to a lesser degree in O-antigen mutants. Aggregates were found to persist in the bloodstream of mice. Importantly, organisms grown under physiological shear were found to have an antibiotic resistance phenotype intermediate between that of fully planktonic and biofilm states. Conclusions. When grown under intravascular-magnitude fluid dynamic conditions, K. pneumoniae spontaneously develops into multicellular aggregates that are capable of persisting in the circulation and exhibit increased antibiotic resistance. PMID:22711903

  1. Adaptive laboratory evolution of Klebsiella pneumoniae for improving 2,3-butanediol production

    PubMed Central

    Li, Hongbiao; Zhang, Genlin; Dang, Yanyan

    2016-01-01

    ABSTRACT Microbial production of 2,3-butanediol is limited by the toxic components in the lignocellulose hydrolysate. To improve the 2,3-butanediol production via Klebsiella pneumoniae from cotton stalk hydrolysate, a method coupling a high tolerance of strain and detoxification of the hydrolysate was thus investigated in this study. The strain tolerance of K. pneumoniae to the cotton stalk hydrolysate was improved via an adaptive laboratory evolution, which involved a stepwise increase in the hydrolysate concentration in the medium. Compared with the initial strain, the resulting strain increased the biomass 3.2-fold in a medium of 20 g/L hydrolysate and produced 10.45 g/L of 2,3-butanediol at an optimal concentration of 60 g/L hydrolysate. After detoxification of cotton stalk hydrolysate, the cell metabolism of K. pneumoniae was further promoted, and the 2,3-butanediol production increased by 1.2 folds. Using fed-batch fermentation, the concentration of 2,3-butanediol reached 35.5 g/L with a yield of 0.43 g/g. The results demonstrated that the bioconversion of low-cost cotton stalk hydrolysate into 2,3-butanediol improves the economics of microbial 2,3-butanediol production. PMID:27442598

  2. Outbreak of KPC-3-producing ST15 and ST348 Klebsiella pneumoniae in a Portuguese hospital.

    PubMed

    Vubil, D; Figueiredo, R; Reis, T; Canha, C; Boaventura, L; DA Silva, G J

    2017-02-01

    To date, only a few sporadic cases of infections due to Klebsiella pneumoniae carbapenemase (KPC) producers have been reported in Portugal. Here, we report for the first time an outbreak of K. pneumoniae KPC-3 producers in a tertiary-care hospital during 2013. Twenty-seven ertapenem-resistant K. pneumoniae were identified in patients at a tertiary-care hospital during 2013 isolated predominantly from urine (48·1%) and blood (25·9%) cultures. All isolates were highly resistant to β-lactam antibiotics and most showed intermediate resistance to imipenem. The more frequent β-lactamases were TEM- (77·7%), CTX-M- (70·3%) and KPC-type (66·6%). KPC-3 was identified by sequencing. The bla KPC-3 gene was associated with an IncF plasmid, and efficiently transferred to E. coli J53. Pulsed-field gel electrophoresis typing revealed three clusters of isolates which were further characterized by multi-locus sequence typing as ST11, ST15 and ST348. Ertapenem-resistant ST15 was already in circulation in the hospital, related to expression of OmpK36 modified porin, but the other two sequence types had not been previously found in the hospital. We conclude that the IncF plasmid mediated transfer of KPC-3 in the outbreak and that implementation of carbapenemase gene screening in isolates from patients on admission to hospital is advisable in order to control dissemination of these antimicrobial resistance elements.

  3. Genomic and transcriptomic analysis of NDM-1 Klebsiella pneumoniae in spaceflight reveal mechanisms underlying environmental adaptability.

    PubMed

    Li, Jia; Liu, Fei; Wang, Qi; Ge, Pupu; Woo, Patrick C Y; Yan, Jinghua; Zhao, Yanlin; Gao, George F; Liu, Cui Hua; Liu, Changting

    2014-08-28

    The emergence and rapid spread of New Delhi Metallo-beta-lactamase-1 (NDM-1)-producing Klebsiella pneumoniae strains has caused a great concern worldwide. To better understand the mechanisms underlying environmental adaptation of those highly drug-resistant K. pneumoniae strains, we took advantage of the China's Shenzhou 10 spacecraft mission to conduct comparative genomic and transcriptomic analysis of a NDM-1 K. pneumoniae strain (ATCC BAA-2146) being cultivated under different conditions. The samples were recovered from semisolid medium placed on the ground (D strain), in simulated space condition (M strain), or in Shenzhou 10 spacecraft (T strain) for analysis. Our data revealed multiple variations underlying pathogen adaptation into different environments in terms of changes in morphology, H2O2 tolerance and biofilm formation ability, genomic stability and regulation of metabolic pathways. Additionally, we found a few non-coding RNAs to be differentially regulated. The results are helpful for better understanding the adaptive mechanisms of drug-resistant bacterial pathogens.

  4. Prolonged delay for controlling KPC-2-producing Klebsiella pneumoniae outbreak: the role of clinical management.

    PubMed

    Delory, T; Seringe, E; Antoniotti, G; Novakova, I; Goulenok, C; Paysant, I; Boyer, S; Carbonne, A; Naas, T; Astagneau, P

    2015-10-01

    Carbapenemase-producing Enterobacteriaceae (CPE) are becoming of immediate concern for infection control policies. Prompt detection of CPE on health care setting admission is crucial to halt the spread of an outbreak. We report a cluster of 13 Klebsiella pneumoniae carbapenemase (KPC)-2-producing K pneumoniae cases in a tertiary care hospital.The objective of this study was to identify contributing factors originating the outbreak. An outbreak investigation was conducted using descriptive epidemiology, observation of health care practices, and interviews of management staff. A root cause analysis was performed to identify patent and latent failures of infection control measures using the association of litigation and risk management method. The main patent failure was the delay in identifying KPC-2-producing K pneumoniae carriers. Contributing factors were work and environmental factors: understaffing, lack of predefined protocols, staff members' characteristics, and underlying patients' characteristics. Latent failures were as follows: no promotion of the national guidelines for prevention of CPE transmission, no clear procedure for the management of patients hospitalized abroad, no clear initiative for promoting a culture of quality in the hospital, biologic activity recently outsourced to a private laboratory, and poor communication among hospital members. Clinical management should be better promoted to control hospital outbreaks and should include team work and safety culture. Copyright © 2015 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

  5. Acquired resistance to innate immune clearance promotes Klebsiella pneumoniae ST258 pulmonary infection

    PubMed Central

    Ahn, Danielle; Peñaloza, Hernán; Wang, Zheng; Wickersham, Matthew; Parker, Dane; Patel, Purvi; Koller, Antonius; Chen, Emily I.; Bueno, Susan M.; Uhlemann, Anne-Catrin; Prince, Alice

    2016-01-01

    Adaptive changes in the genome of a locally predominant clinical isolate of the multidrug-resistant Klebsiella pneumoniae ST258 (KP35) were identified and help to explain the selection of this strain as a successful pulmonary pathogen. The acquisition of 4 new ortholog groups, including an arginine transporter, enabled KP35 to outcompete related ST258 strains lacking these genes. KP35 infection elicited a monocytic response, dominated by Ly6Chi monocytic myeloid-derived suppressor cells that lacked phagocytic capabilities, expressed IL-10, arginase, and antiinflammatory surface markers. In comparison with other K. pneumoniae strains, KP35 induced global changes in the phagocytic response identified with proteomics, including evasion of Ca2+ and calpain activation necessary for phagocytic killing, confirmed in functional studies with neutrophils. This comprehensive analysis of an ST258 K. pneumoniae isolate reveals ongoing genetic adaptation to host microenvironments and innate immune clearance mechanisms that complements its repertoire of antimicrobial resistance genes and facilitates persistence in the lung. PMID:27777978

  6. Expression and purification of functionally active ferrous iron transporter FeoB from Klebsiella pneumoniae.

    PubMed

    Smith, Aaron T; Sestok, Alexandrea E

    2017-09-20

    The acquisition of ferrous iron (Fe(2+)) is an important virulence factor utilized by several hospital-acquired (nosocomial) pathogens such as Klebsiella pneumoniae to establish infection within human hosts. Virtually all bacteria use the ferrous iron transport system (Feo) to acquire ferrous iron from their environments, which are often biological niches that stabilize Fe(2+) relative to Fe(3+). However, the details of this process remain poorly understood, likely owing to the few expression and purification systems capable of supplying sufficient quantities of the chief component of the Feo system, the integral membrane GTPase FeoB. This bottleneck has undoubtedly hampered efforts to understand this system in order to target it for therapeutic intervention. In this study, we describe the expression, solubilization, and purification of the Fe(2+) transporter from K. pneumoniae, KpFeoB. We show that this protein may be heterologously overexpressed in Escherichia coli as the host organism. After testing several different commercially-available detergents, we have developed a solubilization and purification protocol that produces milligram quantities of KpFeoB with sufficient purity for enzymatic and biophysical analyses. Importantly, we demonstrate that KpFeoB displays robust GTP hydrolysis activity (kcat(GTP) of ∼10(-1) s(-1)) in the absence of any additional stimulatory factors. Our findings suggest that K. pneumoniae may be capable of using its Feo system to drive Fe(2+) import in an active manner. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. budC knockout in Klebsiella pneumoniae for bioconversion from glycerol to 1,3-propanediol.

    PubMed

    Guo, Xinkun; Fang, Huiying; Zhuge, Bin; Zong, Hong; Song, Jian; Zhuge, Jian; Du, Xingxing

    2013-01-01

    2,3-Butanediol (2,3-BD) is a major by-product of 1,3-propanediol (1,3-PDO) fermentation by Klebsiella pneumoniae ZG25. It not only consumes large amounts of its carbon source and nicotinamide adenine dinucleotide to diminish synthesis of 1,3-PDO, but also serves as an obstacle to high-purity 1,3-PDO in downstream processes. To decrease the formation of 2,3-BD and make an intrinsic improvement in 1,3-PDO production, the budC gene in K. pneumoniae, coding 2,3-BD dehydrogenase, which is a key gene of the 2,3-BD pathway, was successfully knocked out using the Red recombination system described in this paper. The results of the mutant fed-batch fermentation showed that the 1,3-PDO concentration, productivity per cell dry weight, and conversion rate increased to 880 mmol L(-1) , 22.0 mmol L(-1) h(-1) , and 0.700 mol mol(-1) , respectively, increasing by 10%, 15%, and 11% compared with the parent strain. Meanwhile, 2,3-BD was still found in fermentation broth with the 2,3-BD metabolic pathway blocked, which implies that K. pneumoniae possesses a pathway of the 2,3-BD cycle as a replenishment pathway.

  8. Multiplex PCR for Identification of Two Capsular Types in Epidemic KPC-Producing Klebsiella pneumoniae Sequence Type 258 Strains

    PubMed Central

    Chen, Liang; Chavda, Kalyan D.; Findlay, Jacqueline; Peirano, Gisele; Hopkins, Katie; Pitout, Johann D. D.; Bonomo, Robert A.; Woodford, Neil; DeLeo, Frank R.

    2014-01-01

    We developed a multiplex PCR assay capable of identifying two capsular polysaccharide synthesis sequence types (sequence type 258 [ST258] cps-1 and cps-2) in epidemic Klebsiella pneumoniae ST258 strains. The assay performed with excellent sensitivity (100%) and specificity (100%) for identifying cps types in 60 ST258 K. pneumoniae sequenced isolates. The screening of 419 ST258 clonal isolates revealed a significant association between cps type and K. pneumoniae carbapenemase (KPC) variant: cps-1 is largely associated with KPC-2, while cps-2 is primarily associated with KPC-3. PMID:24733470

  9. Bacteremia and other body site infection caused by hypervirulent and classic Klebsiella pneumoniae.

    PubMed

    Wu, Hua; Li, Dongdong; Zhou, Haijian; Sun, Yunfang; Guo, Ling; Shen, Dingxia

    2017-03-01

    To investigate bacteremia and other body site infection caused by hypervirulent Klebsiella pneumoniae (hvKP), a recently recognized pathogen of invasive infection, and classic Klebsiella pneumoniae (cKP), a very common organism associated with many kinds of nosocomial infection. Clinical information obtained from patients with both bacteremia and other body site infections caused by hvKP and/or cKP was retrospectively reviewed. Homo-hvKP (or homo-cKP) was defined as homologous hvKP (or cKP) strains from different body sites in each individual patient according to string test, virulence gene amplification and PFGE pattern. MLST was carried on to understand the correlation of sequence type with capsular polysaccharide type for Klebsiella pneumoniae from blood. Sixty-four hvKP and 101 cKP strains were isolated from blood and other body sites of 76 patients who had bacteremia accompanied by other site infection. Among these patients, 27 were infected with homo-hvKP, 32 were with homo-cKP, 12 were with heterogeneous cKP, and five were with both hvKP and cKP. Patients with bacteremia and liver abscesses caused by homo-hvKP accounted for 51.9%, and 92.6% of homo-hvKP infected patients did not receive any invasive procedures before bacteremia. However, patients with bacteremia and biliary tract infection caused by homo-cKP accounted for 34.4%, and 78.1% of homo-cKP infected patients had history of invasive procedures before bacteremia. More homo-hvKP strains (59.3%) than homo-cKP strains (34.4%) were isolated from blood earlier than other sites. HvKP strains were statistically more susceptible to the tested antimicrobials than cKP strains. An outbreak of carbapenem-resistant cKP infection and possible gene transfer of KPC-2 from cKP to hvKP were brought to notice. Both hvKP and cKP could cause bacteremia and other body site infection. But patients with hvKP bacteremia usually suffered from liver abscess without previous invasive procedures, most patients with c

  10. Comparative In Vitro Activities of Ciprofloxacin, Clinafloxacin, Gatifloxacin, Levofloxacin, Moxifloxacin, and Trovafloxacin against Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, and Enterobacter aerogenes Clinical Isolates with Alterations in GyrA and ParC Proteins

    PubMed Central

    Brisse, Sylvain; Milatovic, Dana; Fluit, Ad C.; Verhoef, Jan; Martin, Nele; Scheuring, Sybille; Köhrer, Karl; Schmitz, Franz-Josef

    1999-01-01

    The in vitro activities of ciprofloxacin, clinafloxacin, gatifloxacin, levofloxacin, moxifloxacin, and trovafloxacin were tested against 72 ciprofloxacin-resistant and 28 ciprofloxacin-susceptible isolates of Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, and Enterobacter aerogenes. Irrespective of the alterations in GyrA and ParC proteins, clinafloxacin exhibited greater activity than all other fluoroquinolones tested against K. pneumoniae and E. aerogenes. PMID:10428935

  11. Comparative analysis of diguanylate cyclase and phosphodiesterase genes in Klebsiella pneumoniae

    PubMed Central

    2012-01-01

    Background Klebsiella pneumoniae can be found in environmental habitats as well as in hospital settings where it is commonly associated with nosocomial infections. One of the factors that contribute to virulence is its capacity to form biofilms on diverse biotic and abiotic surfaces. The second messenger Bis-(3’-5’)-cyclic dimeric GMP (c-di-GMP) is a ubiquitous signal in bacteria that controls biofilm formation as well as several other cellular processes. The cellular levels of this messenger are controlled by c-di-GMP synthesis and degradation catalyzed by diguanylate cyclase (DGC) and phophodiesterase (PDE) enzymes, respectively. Many bacteria contain multiple copies of these proteins with diverse organizational structure that highlight the complex regulatory mechanisms of this signaling network. This work was undertaken to identify DGCs and PDEs and analyze the domain structure of these proteins in K. pneumoniae. Results A search for conserved GGDEF and EAL domains in three sequenced K. pneumoniae genomes showed that there were multiple copies of GGDEF and EAL containing proteins. Both single domain and hybrid GGDEF proteins were identified: 21 in K. pneumoniae Kp342, 18 in K. pneumoniae MGH 78578 and 17 in K. pneumoniae NTUH-K2044. The majority had only the GGDEF domain, most with the GGEEF motif, and hybrid proteins containing both GGDEF and EAL domains were also found. The I site for allosteric control was identified only in single GGDEF domain proteins and not in hybrid proteins. EAL-only proteins, containing either intact or degenerate domains, were also identified: 15 in Kp342, 15 in MGH 78578 and 10 in NTUH-K2044. Several input sensory domains and transmembrane segments were identified, which together indicate complex regulatory circuits that in many cases can be membrane associated. Conclusions The comparative analysis of proteins containing GGDEF/EAL domains in K. pneumoniae showed that most copies were shared among the three strains and that some

  12. Genomic Characterization of Colistin Heteroresistance in Klebsiella pneumoniae during a Nosocomial Outbreak.

    PubMed

    Halaby, Teysir; Kucukkose, Emre; Janssen, Axel B; Rogers, Malbert R C; Doorduijn, Dennis J; van der Zanden, Adri G M; Al Naiemi, Nashwan; Vandenbroucke-Grauls, Christina M J E; van Schaik, Willem

    2016-11-01

    Klebsiella pneumoniae is emerging as an important nosocomial pathogen due to its rapidly increasing multidrug resistance, which has led to a renewed interest in polymyxin antibiotics, such as colistin, as antibiotics of last resort. However, heteroresistance (i.e., the presence of a subpopulation of resistant bacteria in an otherwise susceptible culture) may hamper the effectiveness of colistin treatment in patients. In a previous study, we showed that colistin resistance among extended-spectrum-beta-lactamase (ESBL)-producing K. pneumoniae isolates emerged after the introduction of selective digestive tract decontamination (SDD) in an intensive care unit (ICU). In this study, we investigated heteroresistance to colistin among ESBL-producing K. pneumoniae isolates by using population analysis profiles (PAPs). We used whole-genome sequencing (WGS) to identify the mutations that were associated with the emergence of colistin resistance in these K. pneumoniae isolates. We found five heteroresistant subpopulations, with colistin MICs ranging from 8 to 64 mg/liter, which were derived from five clonally related, colistin-susceptible clinical isolates. WGS revealed the presence of mutations in the lpxM, mgrB, phoQ, and yciM genes in colistin-resistant K. pneumoniae isolates. In two strains, mgrB was inactivated by an IS3-like or ISKpn14 insertion sequence element. Complementation in trans with the wild-type mgrB gene resulted in these strains reverting to colistin susceptibility. The MICs for colistin-susceptible strains increased 2- to 4-fold in the presence of the mutated phoQ, lpxM, and yciM alleles. In conclusion, the present study indicates that heteroresistant K. pneumoniae subpopulations may be selected for upon exposure to colistin. Mutations in mgrB and phoQ have previously been associated with colistin resistance, but we provide experimental evidence for roles of mutations in the yciM and lpxM genes in the emergence of colistin resistance in K. pneumoniae

  13. Comparative analysis of diguanylate cyclase and phosphodiesterase genes in Klebsiella pneumoniae.

    PubMed

    Cruz, Diana P; Huertas, Mónica G; Lozano, Marcela; Zárate, Lina; Zambrano, María Mercedes

    2012-07-09

    Klebsiella pneumoniae can be found in environmental habitats as well as in hospital settings where it is commonly associated with nosocomial infections. One of the factors that contribute to virulence is its capacity to form biofilms on diverse biotic and abiotic surfaces. The second messenger Bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) is a ubiquitous signal in bacteria that controls biofilm formation as well as several other cellular processes. The cellular levels of this messenger are controlled by c-di-GMP synthesis and degradation catalyzed by diguanylate cyclase (DGC) and phophodiesterase (PDE) enzymes, respectively. Many bacteria contain multiple copies of these proteins with diverse organizational structure that highlight the complex regulatory mechanisms of this signaling network. This work was undertaken to identify DGCs and PDEs and analyze the domain structure of these proteins in K. pneumoniae. A search for conserved GGDEF and EAL domains in three sequenced K. pneumoniae genomes showed that there were multiple copies of GGDEF and EAL containing proteins. Both single domain and hybrid GGDEF proteins were identified: 21 in K. pneumoniae Kp342, 18 in K. pneumoniae MGH 78578 and 17 in K. pneumoniae NTUH-K2044. The majority had only the GGDEF domain, most with the GGEEF motif, and hybrid proteins containing both GGDEF and EAL domains were also found. The I site for allosteric control was identified only in single GGDEF domain proteins and not in hybrid proteins. EAL-only proteins, containing either intact or degenerate domains, were also identified: 15 in Kp342, 15 in MGH 78578 and 10 in NTUH-K2044. Several input sensory domains and transmembrane segments were identified, which together indicate complex regulatory circuits that in many cases can be membrane associated. The comparative analysis of proteins containing GGDEF/EAL domains in K. pneumoniae showed that most copies were shared among the three strains and that some were unique to a particular strain

  14. Structure and Function of CutC Choline Lyase from Human Microbiota Bacterium Klebsiella pneumoniae.

    PubMed

    Kalnins, Gints; Kuka, Janis; Grinberga, Solveiga; Makrecka-Kuka, Marina; Liepinsh, Edgars; Dambrova, Maija; Tars, Kaspars

    2015-08-28

    CutC choline trimethylamine-lyase is an anaerobic bacterial glycyl radical enzyme (GRE) that cleaves choline to produce trimethylamine (TMA) and acetaldehyde. In humans, TMA is produced exclusively by the intestinal microbiota, and its metabolite, trimethylamine oxide, has been associated with a higher risk of cardiovascular diseases. Therefore, information about the three-dimensional structures of TMA-producing enzymes is important for microbiota-targeted drug discovery. We have cloned, expressed, and purified the CutC GRE and the activating enzyme CutD from Klebsiella pneumoniae, a representative of the human microbiota. We have determined the first crystal structures of both the choline-bound and choline-free forms of CutC and have discovered that binding of choline at the ligand-binding site triggers conformational changes in the enzyme structure, a feature that has not been observed for any other characterized GRE.

  15. DHA system mediating aerobic and anaerobic dissimilation of glycerol in Klebsiella pneumoniae NCIB 418.

    PubMed Central

    Forage, R G; Lin, E C

    1982-01-01

    In Klebsiella pneumoniae NCIB 418, the pathways normally responsible for aerobic growth on glycerol and sn-glycerol 3-phosphate (the glp system) are superrepressed. However, aerobic growth on glycerol can take place by the intervention of the NAD-linked glycerol dehydrogenase and the ATP-dependent dihydroxyacetone kinase of the dha system normally inducible only anaerobically by glycerol or dihydroxyacetone. Conclusive evidence that the dha system is responsible for both aerobic and anaerobic dissimilation of glycerol was provided by a Tn5 insertion mutant lacking dihydroxyacetone kinase. An enzymatically coupled assay specific for this enzyme was devised. Spontaneous reactivation of the glp system was achieved by selection for aerobic growth on sn-glycerol 3-phosphate or on limiting glycerol as the sole carbon and energy source. However, the expression of this system became constitutive. Aerobic operation of the glp system highly represses synthesis of the dha system enzymes by catabolite repression. Images PMID:6284704

  16. Klebsiella pneumoniae Spinal Epidural Abscess treated conservatively: case report and review.

    PubMed

    Araújo, Filipe; Ribeiro, Célia; Silva, Inês; Nero, Patrícia; Branco, Jaime C

    2012-01-01

    Spinal infections are rare but potentially life-threatening disorders. A high level of clinical suspicion is necessary for rapid diagnosis and treatment initiation. The treatment combines both antibiotics and surgical intervention in the vast majority of cases. The authors report the case of a 84-year old female patient with a three week history of persistent lumbar back pain radiating to both thighs following a lower respiratory tract infection. She had lumbar spine tenderness but no neurological compromise. Her inflammatory markers were elevated and lumbar spine magnetic resonance imaging revealed L4-L5 spondylodiscitis with spinal epidural abscess. Blood cultures isolated Klebsiella pneumoniae and, since she was neurologically stable, conservative treatment with two-week intravenous gentamicin and eight-week intravenous ceftriaxone was initiated with positive inpatient and outpatient evolution.

  17. Enzymatic evidence for an involvement of pyruvate dehydrogenase in the anaerobic glycerol metabolism of Klebsiella pneumoniae.

    PubMed

    Menzel, K; Zeng, A P; Deckwer, W D

    1997-08-11

    Stoichiometric analysis of pathways involved in anaerobic bioconversion of glycerol by Klebsiella pneumoniae revealed that enzyme(s) in addition to pyruvate formate-lyase (PFL) must be involved in pyruvate decarboxylation. In this work, enzymatic evidence is presented that confirmed a simultaneous involvement of pyruvate dehydrogenase complex (PDH) and excluded the presence of pyruvate:ferredoxin oxidoreductase in this anaerobic bioprocess. The in vitro PDH activity of cell extract from continuous culture was found to be strongly affected by the substrate (glycerol) concentration in medium and cell growth rate (dilution rate). It increases with increasing glycerol concentration and correlates well with the specific substrate uptake rate at different dilution rates in a kind of saturation function. At a similar substrate uptake rate, it decreases with cell growth rate. The in vitro activity of PDH is much higher than its in vivo activity calculated from the pathway stoichiometry but comparable to the calculated in vivo activity of PFL.

  18. Tigecycline therapy for carbapenem-resistant Klebsiella pneumoniae (CRKP) bacteriuria leads to tigecycline resistance.

    PubMed

    van Duin, D; Cober, E D; Richter, S S; Perez, F; Cline, M; Kaye, K S; Kalayjian, R C; Salata, R A; Evans, S R; Fowler, V G; Bonomo, R A

    2014-12-01

    Carbapenem-resistant Klebsiella pneumoniae (CRKP) is an increasing global threat. Here, we describe the prevalence and impact of tigecycline use in a cohort of patients with CRKP bacteriuria nested within a multicentre, prospective study. In the 21-month study period, 260 unique patients were included. Tigecycline was given to 80 (31%) patients. The use of tigecycline during the index hospitalization was significantly associated with the subsequent development of tigecycline resistance in the same patient (OR, 6.13; 95% CI, 1.15-48.65; p 0.03). In conclusion, the use of tigecycline with CRKP bacteriuria is common, and is associated with the subsequent development of tigecycline resistance. © 2014 The Authors Clinical Microbiology and Infection © 2014 European Society of Clinical Microbiology and Infectious Diseases.

  19. Structural and kinetic insights into the mechanism of 5-hydroxyisourate hydrolase from Klebsiella pneumoniae

    SciTech Connect

    French, Jarrod B.; Ealick, Steven E.

    2011-07-19

    The stereospecific oxidative degradation of uric acid to (S)-allantoin has recently been demonstrated to proceed via two unstable intermediates and requires three separate enzymatic reactions. The second step of this reaction, the conversion of 5-hydroxyisourate (HIU) to 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline, is catalyzed by HIU hydrolase (HIUH). The high-resolution crystal structure of HIUH from the opportunistic pathogen Klebsiella pneumoniae (KpHIUH) has been determined. KpHIUH is a homotetrameric protein that, based on sequence and structural similarity, belongs to the transthyretin-related protein family. In addition, the steady-state kinetic parameters for this enzyme and four active-site mutants have been measured. These data provide valuable insight into the functional roles of the active-site residues. Based upon the structural and kinetic data, a mechanism is proposed for the KpHIUH-catalyzed reaction.

  20. First identification of a patient colonized with Klebsiella pneumoniae carrying blaNDM-1 in Taiwan.

    PubMed

    Wu, Hua-Shin; Chen, Te-Li; Chen, Isaac Chun-Jen; Huang, Mu-Shun; Wang, Fu-Der; Fung, Chang-Phone; Lee, Shou-Dong

    2010-11-01

    New Delhi metallo-β-lactamase 1 (NDM-1) is a novel type of metallo-β-lactamase (MBL). Enterobacteriaceae carrying this NDM-1 encoding gene, bla(NDM-1), have been identified worldwide. Bacteria carrying bla(NDM-1) are not only resistant to carbapenem, but also highly resistant to many classes of antibiotics, which indicate the importance of prompt identification of these bacteria and implementation of strict infection control measures to prevent their transmission. Here, we report the first identification and management of a patient colonized with Klebsiella pneumoniae carrying bla(NDM-1) in Taiwan, who returned from New Delhi where he had been hospitalized for a gun-shot injury.

  1. Plasmid-Mediated Antibiotic Resistance and Virulence in Gram-negatives: the Klebsiella pneumoniae Paradigm

    PubMed Central

    Ramirez, Maria S.; Traglia, German M.; Lin, David L.; Tran, Tung; Tolmasky, Marcelo E.

    2015-01-01

    Summary Plasmids harbor genes coding for specific functions including virulence factors and antibiotic resistance that permit bacteria to survive the hostile environment found in the host and resist treatment. Together with other genetic elements such as integrons and transposons, and using a variety of mechanisms, plasmids participate in the dissemination of these traits resulting in the virtual elimination of barriers among different kinds of bacteria. In this article we review the current information about physiology and role in virulence and antibiotic resistance of plasmids from the gram-negative opportunistic pathogen Klebsiella pneumoniae. This bacterium has acquired multidrug resistance and is the causative agent of serious communityand hospital-acquired infections. It is also included in the recently defined ESKAPE group of bacteria that cause most of US hospital infections. PMID:25705573

  2. Polymer production by Klebsiella pneumoniae 4-hydroxyphenylacetic acid hydroxylase genes cloned in Escherichia coli.

    PubMed Central

    Gibello, A; Ferrer, E; Sanz, J; Martin, M

    1995-01-01

    The expression of Klebsiella pneumoniae hpaA and hpaH genes, which code for 4-hydroxyphenylacetic acid hydroxylase in Escherichia coli K-12 derivative strains, is associated with the production of a dark brown pigment in the cultures. This pigment has been identified as a polymer which shows several of the characteristics reported for microbial melanins and results from the oxidative activity of 4-hydroxyphenylacetic acid hydroxylase on some dihydroxylated compounds to form o-quinones. A dibenzoquinone is formed from the oxidation of different mono- or dihydroxylated aromatic compounds by the enzyme prior to polymerization. We report a hydroxylase activity, other than tyrosinase, that is associated with the synthesis of a bacterial melanin. PMID:8534083

  3. Stable chromosomal integration of the entire nitrogen fixation gene cluster from Klebsiella pneumoniae in yeast.

    PubMed Central

    Zamir, A; Maina, C V; Fink, G R; Szalay, A A

    1981-01-01

    A bacterial plasmid containing the entire nitrogen fixation (nif) gene cluster (consisting of at least 15 genes) from Klebsiella pneumoniae was used in conjunction with an Escherichia coli-yeast shuttle plasmid containing the yeast his4 gene cluster to cotransform a his4- recipient strain of Saccharomyces cerevisiae. Of 87 histidine-independent clones screened, 2 contained nif DNA. Restriction and hybridization analyses showed that two copies of the nif plasmid (46 kilobases each) are integrated in tandem in the recipient chromosome by recombination between homologous regions in the transforming plasmids. Chromosomal integration was also verified by tetrad analysis, showing that the nif DNA behaved in meiosis like a Mendelian element. During mitotic growth, one of the two copies of the nif region is frequently lost. The remaining copy of nif is stable, even after 40 generations in nonselective medium. Images PMID:6267596

  4. Septic shock due to Klebsiella pneumoniae after medical abortion with misoprostol-only regimen.

    PubMed

    Kaponis, Apostolos; Papatheodorou, Stefania; Makrydimas, George

    2010-09-01

    To report a case of a healthy woman who was admitted to the hospital with septic shock caused by a common uropathogen after self-administration of misoprostol for pregnancy termination. Case report. Tertiary hospital. A 38-year-old woman, gravida 5, para 3, who developed septic shock after medical termination of pregnancy. Suction curettage, antibiotic treatment, plasma and platelet transfusions. Klebsiella pneumoniae was isolated from blood samples. Ten days after her admission she was discharged home in good condition on oral antibiotics. Severe infections leading to septic shock from common pathogen bacteria can occur after medical termination of pregnancy, independently of the regimen used. Copyright (c) 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  5. High Prevalence of ESBL-Producing Klebsiella pneumoniae Causing Community-Onset Infections in China

    PubMed Central

    Zhang, Jing; Zhou, Kai; Zheng, Beiwen; Zhao, Lina; Shen, Ping; Ji, Jinru; Wei, Zeqing; Li, Lanjuan; Zhou, Jianying; Xiao, Yonghong

    2016-01-01

    The aim of this work was to investigate the epidemiological and genetic characteristics of ESBL-producing Klebsiella pneumoniae (ESBL-Kp) causing community-onset infections. K. pneumoniae isolates were collected from 31 Chinese secondary hospitals between August 2010 and 2011. Genes encoding ESBL and AmpC beta-lactamases were detected by PCR. The isolates were assigned to sequence types (STs) using multi-locus sequence typing (MLST). Eleven ESBL-Kp strains were selected for whole-genome sequencing (WGS) for investigating the genetic environment and plasmids encoding ESBL genes. A total of 578 K. pneumoniae isolates were collected, and 184 (31.8%) carried ESBL genes. The prevalence of ESBL-Kp varied from different geographical areas of China (10.2–50.3%). The three most prevalent ESBL genes were blaCTX-M-14 (n = 74), blaCTX-M-15 (n = 60), and blaCTX-M-3 (n = 40). MLST assigned 127 CTX-M-14 and CTX-M-15 producers to 54 STs, and CC17 was the most prevalent population (12.6%). STs (23, 37, and 86) that were known frequently associated with hypervirulent K. pneumoniae (hvKP) account for 14.1% (18/127). Phylogenetic analysis by concatenating the seven loci of MLST revealed the existence of ESBL-producing K. quasipneumoniae (two strains) and K. varricola (one strain), which was further confirmed by WGS. This study highlights the challenge of community-onset infections caused by ESBL-Kp in China. The prevalence of STs frequently associating with hvKP should be of concern. Surveillance of ESBL-KP causing community-onset infections now appears imperative. PMID:27895637

  6. Clonal Dissemination of OXA-370-Producing Klebsiella pneumoniae in Rio de Janeiro, Brazil.

    PubMed

    Pereira, Polyana Silva; Borghi, Mirla; de Araújo, Carlos Felipe Machado; Aires, Caio Augusto Martins; Oliveira, Jane Cleide Ribeiro; Asensi, Marise Dutra; Carvalho-Assef, Ana Paula D'Alincourt

    2015-08-01

    Enzymes of the OXA-48 family have become some of the most important beta-lactamases in the world. A new OXA-48 variant (OXA-370) was first described for an Enterobacter hormaechei strain isolated in Rio Grande do Sul (southern region of Brazil) in 2013. Here we report detection of the blaOXA-370 gene in 24 isolates belonging to three Enterobacteriaceae species (22 Klebsiella pneumoniae isolates, 1 Enterobacter cloacae isolate, and 1 Enterobacter aerogenes isolate) collected from five hospitals in Rio de Janeiro, Brazil, in 2013 and 2014. The isolates showed a multidrug resistance profile, and 12.5% were resistant to polymyxin B. Besides blaOXA-370, no other carbapenemase genes were observed by PCR, whereas blaOXA-1 was found in all isolates and 22 isolates (91.6%) possessed blaCTX-M-15. Molecular typing of the K. pneumoniae isolates by pulsed-field gel electrophoresis (PFGE) showed the presence of two clonal groups, i.e., KpA (21 isolates) and KpB (1 isolate). KpA was characterized as sequence type 16 (ST16) and KpB as ST1041 by multilocus sequence typing (MLST). ST16 has been observed for KPC-producing K. pneumoniae in Rio de Janeiro. Plasmid analysis performed with six representative OXA-370-producing isolates showed plasmids harboring the blaOXA-370 gene in all strains, ranging from 25 kb to 150 kb. This study suggests that there is an urgent need to investigate the presence of OXA-370 and dissemination of the K. pneumoniae ST16 clone carrying this gene in Brazil.

  7. Delay in Human Neutrophil Constitutive Apoptosis after Infection with Klebsiella pneumoniae Serotype K1

    PubMed Central

    Lee, Chen-Hsiang; Chuah, Seng-Kee; Tai, Wei-Chen; Chang, Chia-Chi; Chen, Fang-Ju

    2017-01-01

    Klebsiella pneumoniae serotype K1 is a major cause of invasive syndrome defined by liver abscess with metastatic infections at other body sites. This culprit is known to be resistant to neutrophil phagocytosis and bactericidal activity. We hypothesized that K. pneumoniae serotype K1 might regulate neutrophil apoptosis and enhance the survival of the infected neutrophils that might serve as a vector for dissemination of the bacteria. Two serotypes of K. pneumoniae, KP-M1 isolated from a patient with liver abscess and DT-X (an acapsular mutant strain of KP-M1), were used to infect human neutrophils. The infected neutrophils were examined for their cytotoxicity, annexin V staining, proteins, DNA fragmentation, cytokine production, and viability that are involved in apoptosis. We found that KP-M1 was not destroyed and the ingested bacteria survived within neutrophils. While the uninfected neutrophils became apoptotic within 10 h, the neutrophils infected with KP-M1 could survive up to 24 h post infection. Constitutive apoptosis of KP-M1-infected neutrophils was significantly delayed compared to that of DT-X-infected or uninfected neutrophils (p < 0.01). KP-M1 modulated the anti-apoptotic effects by down-regulating the ratio of Bax to Bcl-2 and Mcl-1, and then delayed caspase-3 activation in the neutrophils, which was accompanied by inducing the anti-apoptotic cytokine, IL-8. These data suggest that K. pneumoniae serotype K1 can prolong the lifespan of infected neutrophils by delaying constitutive apoptosis within the first several hours of infection. PMID:28396849

  8. A nanomechanical study of the effects of colistin on the Klebsiella pneumoniae AJ218 capsule.

    PubMed

    Mularski, Anna; Wilksch, Jonathan; Hanssen, Eric; Li, Jian; Tomita, Takehiro; Pidot, Sacha James; Stinear, Tim; Separovic, Frances; Strugnell, Dick

    2017-05-01

    Atomic force microscopy measurements of capsule thickness revealed that that the wild-type Klebsiella pneumoniae AJ218 capsular polysaccharides were rearranged by exposure to colistin. The increase in capsule thickness measured near minimum inhibitory/bactericidal concentration (MIC/MBC) is consistent with the idea that colistin displaces the divalent cations that cross-bridge adjacent lipopolysaccharide (LPS) molecules through the capsule network. Cryo-electron microscopy demonstrated that the measured capsule thickness at near MIC/MBC of 1.2 μM was inflated by the disrupted outer membrane, through which the capsule is excreted and LPS is bound. Since wild-type and capsule-deficient strains of K. pneumoniae AJ218 have equivalent MICs and MBCs, the presence of the capsule appeared to confer no protection against colistin in AJ218. A spontaneously arising colistin mutant showed a tenfold increase in resistance to colistin; genetic analysis identified a single amino acid substitution (Q95P) in the PmrB sensor kinase in this colistin-resistant K. pneumoniae AJ218. Modification of the lipid A component of the LPS could result in a reduction of the net-negative charge of the outer membrane, which could hinder binding of colistin to the outer membrane and displacement of the divalent cations that bridge adjacent LPS molecules throughout the capsular polysaccharide network. Retention of the cross-linking divalent cations may explain why measurements of capsule thickness did not change significantly in the colistin-resistant strain after colistin exposure. These results contrast with those for other K. pneumoniae strains that suggest that the capsule confers colistin resistance.

  9. Clonal Dissemination of OXA-370-Producing Klebsiella pneumoniae in Rio de Janeiro, Brazil

    PubMed Central

    Pereira, Polyana Silva; Borghi, Mirla; de Araújo, Carlos Felipe Machado; Aires, Caio Augusto Martins; Oliveira, Jane Cleide Ribeiro; Asensi, Marise Dutra

    2015-01-01

    Enzymes of the OXA-48 family have become some of the most important beta-lactamases in the world. A new OXA-48 variant (OXA-370) was first described for an Enterobacter hormaechei strain isolated in Rio Grande do Sul (southern region of Brazil) in 2013. Here we report detection of the blaOXA-370 gene in 24 isolates belonging to three Enterobacteriaceae species (22 Klebsiella pneumoniae isolates, 1 Enterobacter cloacae isolate, and 1 Enterobacter aerogenes isolate) collected from five hospitals in Rio de Janeiro, Brazil, in 2013 and 2014. The isolates showed a multidrug resistance profile, and 12.5% were resistant to polymyxin B. Besides blaOXA-370, no other carbapenemase genes were observed by PCR, whereas blaOXA-1 was found in all isolates and 22 isolates (91.6%) possessed blaCTX-M-15. Molecular typing of the K. pneumoniae isolates by pulsed-field gel electrophoresis (PFGE) showed the presence of two clonal groups, i.e., KpA (21 isolates) and KpB (1 isolate). KpA was characterized as sequence type 16 (ST16) and KpB as ST1041 by multilocus sequence typing (MLST). ST16 has been observed for KPC-producing K. pneumoniae in Rio de Janeiro. Plasmid analysis performed with six representative OXA-370-producing isolates showed plasmids harboring the blaOXA-370 gene in all strains, ranging from 25 kb to 150 kb. This study suggests that there is an urgent need to investigate the presence of OXA-370 and dissemination of the K. pneumoniae ST16 clone carrying this gene in Brazil. PMID:25987619

  10. High Prevalence of ESBL-Producing Klebsiella pneumoniae Causing Community-Onset Infections in China.

    PubMed

    Zhang, Jing; Zhou, Kai; Zheng, Beiwen; Zhao, Lina; Shen, Ping; Ji, Jinru; Wei, Zeqing; Li, Lanjuan; Zhou, Jianying; Xiao, Yonghong

    2016-01-01

    The aim of this work was to investigate the epidemiological and genetic characteristics of ESBL-producing Klebsiella pneumoniae (ESBL-Kp) causing community-onset infections. K. pneumoniae isolates were collected from 31 Chinese secondary hospitals between August 2010 and 2011. Genes encoding ESBL and AmpC beta-lactamases were detected by PCR. The isolates were assigned to sequence types (STs) using multi-locus sequence typing (MLST). Eleven ESBL-Kp strains were selected for whole-genome sequencing (WGS) for investigating the genetic environment and plasmids encoding ESBL genes. A total of 578 K. pneumoniae isolates were collected, and 184 (31.8%) carried ESBL genes. The prevalence of ESBL-Kp varied from different geographical areas of China (10.2-50.3%). The three most prevalent ESBL genes were blaCTX-M-14 (n = 74), blaCTX-M-15 (n = 60), and blaCTX-M-3 (n = 40). MLST assigned 127 CTX-M-14 and CTX-M-15 producers to 54 STs, and CC17 was the most prevalent population (12.6%). STs (23, 37, and 86) that were known frequently associated with hypervirulent K. pneumoniae (hvKP) account for 14.1% (18/127). Phylogenetic analysis by concatenating the seven loci of MLST revealed the existence of ESBL-producing K. quasipneumoniae (two strains) and K. varricola (one strain), which was further confirmed by WGS. This study highlights the challenge of community-onset infections caused by ESBL-Kp in China. The prevalence of STs frequently associating with hvKP should be of concern. Surveillance of ESBL-KP causing community-onset infections now appears imperative.

  11. The IκB family member Bcl-3 coordinates the pulmonary defense against Klebsiella pneumoniae infection.

    PubMed

    Pène, Frédéric; Paun, Andrea; Sønder, Søren Ulrik; Rikhi, Nimisha; Wang, Hongshan; Claudio, Estefania; Siebenlist, Ulrich

    2011-02-15

    Bcl-3 is an atypical member of the IκB family that has the potential to positively or negatively modulate nuclear NF-κB activity in a context-dependent manner. Bcl-3's biologic impact is complex and includes roles in tumorigenesis and diverse immune responses, including innate immunity. Bcl-3 may mediate LPS tolerance, suppressing cytokine production, but it also seems to contribute to defense against select systemic bacterial challenges. However, the potential role of Bcl-3 in organ-specific host defense against bacteria has not been addressed. In this study, we investigated the relevance of Bcl-3 in a lung challenge with the Gram-negative pathogen Klebsiella pneumoniae. In contrast to wild-type mice, Bcl-3-deficient mice exhibited significantly increased susceptibility toward K. pneumoniae pneumonia. The mutant mice showed increased lung damage marked by neutrophilic alveolar consolidation, and they failed to clear bacteria in lungs, which correlated with increased bacteremic dissemination. Loss of Bcl-3 incurred a dramatic cytokine imbalance in the lungs, which was characterized by higher levels of IL-10 and a near total absence of IFN-γ. Moreover, Bcl-3-deficient mice displayed increased lung production of the neutrophil-attracting chemokines CXCL-1 and CXCL-2. Alveolar macrophages and neutrophils are important to antibacterial lung defense. In vitro stimulation of Bcl-3-deficient alveolar macrophages with LPS or heat-killed K. pneumoniae recapitulated the increase in IL-10 production, and Bcl-3-deficient neutrophils were impaired in intracellular bacterial killing. These findings suggest that Bcl-3 is critically involved in lung defense against Gram-negative bacteria, modulating functions of several cells to facilitate efficient clearance of bacteria.

  12. KPC-Producing Klebsiella pneumoniae Isolates in Croatia: A Nationwide Survey.

    PubMed

    Jelic, Marko; Butic, Iva; Plecko, Vanda; Cipris, Ivan; Jajic, Ines; Bejuk, Danijela; Koscak, Iva; Marinkovic, Sonja; Pal, Marina Payerl; Andrasevic, Arjana Tambic

    2016-12-01

    In the last few years, Klebsiella pneumoniae strains producing K. pneumoniae carbapenemase (KPC) enzymes have emerged as important multidrug-resistant pathogens in hospitalized patients. This report describes KPC-producing isolates collected through the Croatian antimicrobial resistance surveillance program in the early stage of their dissemination in Croatia. Forty-eight KPC-producing K. pneumoniae isolates, collected during a period from February 2011 to August 2013, were analyzed in this study. Antimicrobial susceptibility profiles were determined using disk diffusion and E-test. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were used for epidemiological analysis. Identification of β-lactamase genes and associated antibiotic resistance mechanisms was performed by polymerase chain reaction and positive products were sequenced. Localization of blaKPC was investigated by S1 PFGE and Southern hybridization. Of 40 participating centers in Croatia, KPC isolates were recorded in 9 of them. They all had multidrug-resistant phenotype, but showed varying levels of resistance to carbapenems. All isolates displayed ST258, and PFGE showed that all but one were closely related. All isolates harbored blaKPC-2. Isolate with a unique PFGE pattern produced TEM-1, while others produced TEM-116. All isolates harbored blaSHV-11, but were negative for blaCTX-M and blaAmpC genes. All isolates contain one KPC-harboring plasmid, ranging in size from ∼60 to ∼210 kb, characterized as FIIs and IncR. This report describes that the early stage of KPC-producing K. pneumoniae dissemination in Croatia is associated with a prolific PFGE type belonging to ST258. So far, the spread of an outbreak strain is limited to the northwest region of the country.

  13. Murine Immunoprotective Activity of Klebsiella pneumoniae Cell Surface Preparations: Comparative Study with Ribosomal Preparations

    PubMed Central

    Fournier, Jean-Michel; Jolivet-Reynaud, Colette; Riottot, Marie-Madeleine; Jouin, Hélène

    1981-01-01

    Cell surface preparations and ribosomal preparations were extracted from Klebsiella pneumoniae. Agar gel diffusion with antisera to cell surface preparations or ribosomal preparations indicated common antigenic components among the preparations. Lipopolysaccharide and capsular polysaccharide were identified in the cell surface preparations. These results and the previous identification of lipopolysaccharide and capsular polysaccharide in ribosomal preparations suggest that these antigens are responsible for the immunochemical cross-reactivity observed among these two bacterial extracts. Active protection could be induced in mice by these two preparations. On a dry-weight basis, cell surface preparations provided better immunoprotective activity than did ribosomal preparations. However, the 50% protective dose of both preparations is practically the same on the basis of their capsular polysaccharide content. These results are consistent with the hypothesis that the immunoprotective moiety of ribosomal preparations is the contaminating cell surface antigens. Furthermore, the low level of nucleotidic components detected in purified cell surface preparations led us to infer that the immunoprotective activity of capsular polysaccharide may not be dependent on the adjuvant activity of ribonucleic acid. The involvement of capsular polysaccharide in the immunoprotective capacity of cell surface preparations is demonstrated either by using a degradation of this antigen by K. pneumoniae bacteriophage K2-associated glycanase or by using a preparation extracted from a noncapsulated mutant of K. pneumoniae. Nevertheless, the low protective ability of purified capsular polysaccharides is in contrast to its greater activity when induced in bacterial cell surface preparations. The protective activity of K. pneumoniae capsular polysaccharide may be dependent on its association with other surface antigenic components present in cell surface preparations or may be dependent on its

  14. Unravelling of a mechanism of resistance to colistin in Klebsiella pneumoniae using atomic force microscopy.

    PubMed

    Formosa, C; Herold, M; Vidaillac, C; Duval, R E; Dague, E

    2015-08-01

    In this study we focused on the mechanism of colistin resistance in Klebsiella pneumoniae. We used two strains of K. pneumoniae: a colistin-susceptible strain (K. pneumoniae ATCC 700603, KpATCC) and its colistin-resistant derivative (KpATCCm, MIC of colistin 16 mg/L). We performed a genotypic analysis based on the expression of genes involved in LPS synthesis and L-Ara4N moiety addition. We also explored the status of the mgrB gene. Then, a phenotypic analysis was performed using atomic force microscopy (AFM). The Young modulus was extracted from force curves fitted using the Hertz model, and stiffness values were extracted from force curves fitted using the Hooke model. We failed to observe any variation in the expression of genes implicated in LPS synthesis or L-Ara4N moiety addition in KpATCCm, in the absence of colistin or under colistin pressure (versus KpATCC). This led us to identify an insertional inactivation/mutation in the mgrB gene of KpATCCm. In addition, morphology results obtained by AFM showed that colistin removed the capsule from the susceptible strain, but not from the resistant strain. Nanomechanical data on the resistant strain showed that colistin increased the Young modulus of the capsule. Extend force curves recorded on top of the cells allowed us to make the following hypothesis about the nanoarchitecture of the capsule of the two strains: KpATCC has a soft capsule consisting of one layer, whereas the KpATCCm capsule is harder and organized in several layers. We hypothesize that capsular polysaccharides might be implicated in the mechanism of colistin resistance in K. pneumoniae, depending on its genotype. © The Author 2015. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  15. Identification of Outer Membrane and Exoproteins of Carbapenem-Resistant Multilocus Sequence Type 258 Klebsiella pneumoniae

    PubMed Central

    Brinkworth, Amanda J.; Hammer, Carl H.; Olano, L. Renee; Kobayashi, Scott D.; Chen, Liang; Kreiswirth, Barry N.; DeLeo, Frank R.

    2015-01-01

    Carbapenem-resistant Klebsiella pneumoniae strains have emerged as a cause of life-threatening infections in susceptible individuals (e.g., transplant recipients and critically ill patients). Strains classified as multilocus sequence type (ST) 258 are among the most prominent causes of carbapenem-resistant K. pneumoniae infections worldwide, but the basis for the success of this lineage remains incompletely determined. To gain a more comprehensive view of the molecules potentially involved in the success of ST258, we used a proteomics approach to identify surface-associated and culture supernatant proteins produced by ST258. Protein samples were prepared from varied culture conditions in vitro, and were analyzed by a combination of two-dimensional electrophoresis and liquid chromatography followed by tandem mass spectrometry (LC-MS/MS). We identified a total of 193 proteins in outer membrane preparations from bacteria cultured in Luria-Bertani broth (LB) or RPMI 1640 tissue culture media (RPMI). Compared with LB, several iron-acquisition proteins, including IutA, HmuR, HmuS, CirA, FepA, FitA, FoxA, FhuD, and YfeX, were more highly expressed in RPMI. Of the 177 proteins identified in spent media, only the fimbrial subunit, MrkA, was predicted to be extracellular, a finding that suggests few proteins (or a limited quantity) are freely secreted by ST258. Notably, we discovered 203 proteins not reported in previous K. pneumoniae proteome studies. In silico modeling of proteins with unknown function revealed several proteins with beta-barrel transmembrane structures typical of porins, as well as possible host-interacting proteins. Taken together, these findings contribute several new targets for the mechanistic study of drug-resistance and pathogenesis by ST258 K. pneumoniae isolates. PMID:25893665

  16. First Report of bla CTX-M-15-Type ESBL-Producing Klebsiella pneumoniae in Wild Migratory Birds in Pakistan.

    PubMed

    Raza, Shahbaz; Mohsin, Mashkoor; Madni, Waqas Ahmed; Sarwar, Fatima; Saqib, Muhammad; Aslam, Bilal

    2017-01-11

    We investigated wild migratory birds faecal swabs for extended-spectrum β-lactamases-producing Klebsiella pneumoniae (ESBL-K. pneumoniae) from wetland habitats in Pakistan. ESBL-K. pneumoniae were analysed for MDR phenotype, ESBL genotype and genetic diversity. A total of 13 (8.6%) ESBL-K. pneumoniae were recovered. Of these, 8 (61%) isolates were MDR. DNA sequencing confirmed bla CTX-M-15 as the dominant ESBL genotype. BOX-PCR fingerprints showed most of the isolates are unrelated. This study is the first to report the wildlife contamination of CTX-M-15-producing K. pneumoniae in Pakistan. Due to long-range migration, these birds could be responsible for trans-boundary spread of multidrug-resistant bacteria.

  17. Approach to Carbapenemase Detection in Klebsiella pneumoniae in Routine Diagnostic Laboratories.

    PubMed

    Aseem, Rangnekar; Shenoy, Shalini; Mala, Suchitra Shenoy; Baliga, Shrikala; Ashish, Agarwal

    2016-12-01

    Resistance to Carbapenems in Klebsiella may be due to Carbapenem hydrolysing enzymes. Accurate detection of carbapenemase must be done for patient treatment and epidemiological purposes. To detect carbapenemase production by performing Modified Hodge Test (MHT), Combined Disk Test (CDT) for Metallo-β-Lactamases (MBL) and PCR for blaKPC gene, to evaluate the performance of MHT using MacConkey Agar (MCA) and to access the value of MHT for carbapenemase detection. Using a prospective laboratory study design, 153 Extended Spectrum Beta-Lactamases (ESBL) producing Klebsiella pneumoniae from clinical samples of patients admitted in the Kasturba Medical College were collected from January 2014 to December 2015. Isolates resistant to carbapenems by disk diffusion were subjected to MHT on MCA and Mueller Hinton agar (MHA). All isolates were tested for (MBL) production by Imipenem and Imipenem-EDTA CDT and subjected to PCR for the presence of blaKPC gene. Out of 153 isolates, 54 were resistant to one of the carbapenems. Among these, 13 were positive for MHT on MHA, while 23 were positive by MHT on MCA. Number of MBL producers was 23 (42.5%), while blaKPC was detected in 2 out of the 54 isolates. Though detection of drug resistance gene remains the method of choice, it can be performed only in centers with adequate resources. Hence, for most laboratories in resource poor countries, the MHT performed on MCA with concomitant CDT for MBL detection seem to be a better option for detection of Carbapenem resistance.

  18. Antibody-Based Immunotherapy To Treat and Prevent Infection with Hypervirulent Klebsiella pneumoniae.

    PubMed

    Diago-Navarro, Elizabeth; Calatayud-Baselga, Isabel; Sun, Donglei; Khairallah, Camille; Mann, Inderjit; Ulacia-Hernando, Amaia; Sheridan, Brian; Shi, Meiqing; Fries, Bettina C

    2017-01-01

    Hypervirulent Klebsiella pneumoniae (hvKp) strains are predicted to become a major threat in Asia if antibiotic resistance continues to spread. Anticapsular antibodies (Abs) were developed because disseminated infections caused by hvKp are associated with significant morbidity and mortality, even with antibiotic-sensitive strains. K1-serotype polysaccharide capsules (K1-CPS) are expressed by the majority of hvKp strains. In this study, K1-CPS-specific IgG Abs were generated by conjugation of K1-CPS to immunogenic anthrax protective antigen (PA) protein. Opsonophagocytic efficacy was measured in vitro and in vivo by intravital microscopy in murine livers. In vivo protection was tested in murine models, including a novel model for dissemination in hvKp-colonized mice. Protective efficacy of monoclonal antibodies (MAbs) 4C5 (IgG1) and 19A10 (IgG3) was demonstrated both in murine sepsis and pulmonary infection. In hvKp-colonized mice, MAb treatment significantly decreased dissemination of hvKp from the gut to mesenteric lymph nodes and organs. Intravital microscopy confirmed efficient opsonophagocytosis and clearance of bacteria from the liver. In vitro studies demonstrate that MAbs work predominantly by promoting FcR-mediated phagocytosis but also indicate that MAbs enhance the release of neutrophil extracellular traps (NETs). In anticipation of increasing antibiotic resistance, we propose further development of these and other Klebsiella-specific MAbs for therapeutic use. Copyright © 2017 American Society for Microbiology.

  19. Genomic identification of nitrogen-fixing Klebsiella variicola, K. pneumoniae and K. quasipneumoniae.

    PubMed

    Chen, Mingyue; Li, Yuanyuan; Li, Shuying; Tang, Lie; Zheng, Jingwu; An, Qianli

    2016-01-01

    It was difficult to differentiate Klebsiella pneumoniae, K. quasipneumoniae and K. variicola by biochemical and phenotypic tests. Genomics increase the resolution and credibility of taxonomy for closely-related species. Here, we obtained the complete genome sequence of the K. variicola type strain DSM 15968(T) (=F2R9(T)). The genome of the type strain is a circular chromosome of 5,521,203 bp with 57.56% GC content. From 540 Klebsiella strains whose genomes had been publicly available as at 3 March 2015, we identified 21 strains belonging to K. variicola and 8 strains belonging to K. quasipneumoniae based on the genome average nucleotide identities (ANI). All the K. variicola strains, one K. pneumoniae strain and five K. quasipneumoniae strains contained nitrogen-fixing genes. A phylogenomic analysis showed clear species demarcations for these nitrogen-fixing bacteria. In accordance with the key biochemical characteristics of K. variicola, the idnO gene encoding 5-keto-D-gluconate 5-reductase for utilization of 5-keto-D-gluconate and the sorCDFBAME operon for catabolism of L-sorbose were present whereas the rbtRDKT operon for catabolism of adonitol was absent in the genomes of K. variicola strains. Therefore, the genomic analyses supported the ANI-based species delineation; the genome sequence of the K. variicola type strain provides the reference genome for genomic identification of K. variicola, which is a nitrogen-fixing species. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Molecular epidemiology and virulence factors of pyogenic liver abscess causing Klebsiella pneumoniae in China.

    PubMed

    Luo, Y; Wang, Y; Ye, L; Yang, J

    2014-11-01

    The molecular epidemiology and prevalence of virulence factors of isolates from patients with Klebsiella pneumoniae liver abscess (KLA) in mainland China are unknown. Klebsiella pneumoniae isolates were obtained from drainage samples aseptically collected from patients with pyogenic liver abscess (PLA). The genetic similarity of KLA isolates was analyzed by pulsed-field gel electrophoresis. The hypermucoviscosity (HV) phenotype was identified by a positive string test. The K1 and K2 genotypes, the pLVPK-derived genetic loci, aerobactin gene, kfu and alls were detected by PCR amplification. The sequence types (STs) were identified by multilocus sequence typing. Among the 51 non-repetitive KLA isolates, 49 PFGE types have been identified. In total, 19 (37.2%) and 14 (27.4%) of the 51 KLA isolates belonged to clonal complex (CC) 23 and CC65, respectively, while the other 18 isolates (35.3%) were defined as other STs. CC23 consisted of only K1 strains, while CC65 included only K2 strains. All non-K1/K2 strains were classified as STs other than CC23 and CC65. Approximately 70.6% (36/51) of KLA isolates exhibited an HV phenotype. Both K1 and K2 isolates presented significantly higher prevalence of the pLVPK-derived loci than non-K1/K2 isolates. The K1 isolates had a significantly higher prevalence of the kfu and allS genes than K2 and non-K1/K2 isolates, while the K2 isolates exhibited higher repA prevalence than K1 and non-K1/K2 isolates. The majority of KLA isolates belonged to CC23K1 and CC65K2, while other STs with non-K1/K2 capsular types have also been identified. The virulent factors exhibited diverse distribution among the different clones of KLA isolates.

  1. Molecular mechanisms of β-lactam resistance in carbapenemase-producing Klebsiella pneumoniae from Sri Lanka.

    PubMed

    Hall, Jarrad M; Corea, Enoka; Sanjeewani, H D Anusha; Inglis, Timothy J J

    2014-08-01

    Carbapenemases are increasingly important antimicrobial resistance determinants. Little is known about the carbapenem resistance mechanisms in Sri Lanka. We examined 22 carbapenem-resistant Klebsiella pneumoniae from Sri Lanka to determine their β-lactam resistance mechanisms. The predominant resistance mechanisms we detected in this study were OXA-181, NDM-1 carbapenemases and extended-spectrum β-lactamase CTX-M-15. All isolates were then genotyped by pulsed-field gel electrophoresis, variable-number tandem repeat sequence analysis and multilocus sequence typing, and seven distinct genotypes were observed. Five OXA-181-positive Klebsiella pneumoniae isolates were genotypically related to an isolate of Indian origin. Multilocus sequence typing found that these related isolates belong to ST-14, which has been associated with dissemination of OXA-181 from the Indian subcontinent. Other genotypes we discovered were ST-147 and ST-340, also associated with intercontinental spread of carbapenemases of suspected subcontinental origin. The major porin genes ompK35 and ompK36 from these isolates had insertions, deletions and substitutions. Some of these were exclusive to strains within single pulsotypes. We detected one ompK36 variant, ins AA134-135GD, in six ST-14- and six ST-147, blaOXA-181-positive isolates. This porin mutation was an independent predictor of high-level meropenem resistance in our entire Sri Lankan isolate collection (P=0.0030). Analysis of the Sri Lankan ST-14 and ST-147 ins AA134-135GD-positive isolates found ST-14 was more resistant to meropenem than other isolates (mean MIC: 32±0 µg ml(-1) and 20±9.47 µg ml(-1), respectively, P=0.0277). The likely international transmission of these carbapenem resistance determinants highlights the need for regional collaboration and prospective surveillance of carbapenem-resistant Enterobacteriaceae.

  2. Rapid Induction of High-Level Carbapenem Resistance in Heteroresistant KPC-Producing Klebsiella pneumoniae

    PubMed Central

    Adams-Sapper, Sheila; Nolen, Shantell; Donzelli, Grace Fox; Lal, Mallika; Chen, Kunihiko; Justo da Silva, Livia Helena; Moreira, Beatriz M.

    2015-01-01

    Enterobacteriaceae strains producing the Klebsiella pneumoniae carbapenemase (KPC) have disseminated worldwide, causing an urgent threat to public health. KPC-producing strains often exhibit low-level carbapenem resistance, which may be missed by automated clinical detection systems. In this study, eight Klebsiella pneumoniae strains with heterogeneous resistance to imipenem were used to elucidate the factors leading from imipenem susceptibility to high-level resistance as defined by clinical laboratory testing standards. Time-kill analysis with an inoculum as low as 3 × 106 CFU/ml and concentrations of imipenem 8- and 16-fold higher than the MIC resulted in the initial killing of 99.9% of the population. However, full recovery of the population occurred by 20 h of incubation in the same drug concentrations. Population profiles showed that recovery was mediated by a heteroresistant subpopulation at a frequency of 2 × 10−7 to 3 × 10−6. Samples selected 2 h after exposure to imipenem were as susceptible as the unexposed parental strain and produced the major outer membrane porin OmpK36. However, between 4 to 8 h after exposure, OmpK36 became absent, and the imipenem MIC increased at least 32-fold. Individual colonies isolated from cultures after 20 h of exposure revealed both susceptible and resistant subpopulations. Once induced, however, the high-level imipenem resistance was maintained, and OmpK36 remained unexpressed even without continued carbapenem exposure. This study demonstrates the essential coordination between blaKPC and ompK36 expression mediating high-level imipenem resistance from a population of bacteria that initially exhibits a carbapenem-susceptibility phenotype. PMID:25801565

  3. Derepression of Mineral Phosphate Solubilization Phenotype by Insertional Inactivation of iclR in Klebsiella pneumoniae.

    PubMed

    Rajput, Mahendrapal Singh; Iyer, Bhagya; Pandya, Maharshi; Jog, Rahul; G, Naresh Kumar; Rajkumar, Shalini

    2015-01-01

    The mode of succinate mediated repression of mineral phosphate solubilization and the role of repressor in suppressing phosphate solubilization phenotype of two free-living nitrogen fixing Klebsiella pneumoniae strains was studied. Organic acid mediated mineral phosphate solubilization phenotype of oxalic acid producing Klebsiella pneumoniae SM6 and SM11 were transcriptionally repressed by IclR in presence of succinate as carbon source. Oxalic acid production and expression of genes of the glyoxylate shunt (aceBAK) was found only in glucose but not in succinate- and glucose+succinate-grown cells. IclR, repressor of aceBAK operon, was inactivated using an allelic exchange system resulting in derepressed mineral phosphate solubilization phenotype through constitutive expression of the glyoxylate shunt. Insertional inactivation of iclR resulted in increased activity of the glyoxylate shunt enzymes even in succinate-grown cells. An augmented phosphate solubilization up to 54 and 59% soluble phosphate release was attained in glucose+succinate-grown SM6Δ and SM11Δ strains respectively, compared to glucose-grown cells, whereas phosphate solubilization was absent or negligible in wildtype cells grown in glucose+succinate. Both wildtype and iclR deletion strains showed similar indole-3-acetic acid production. Wheat seeds inoculated with wildtype SM6 and SM11 improved both root and shoot length by 1.2 fold. However, iclR deletion SM6Δ and SM11Δ strains increased root and shoot length by 1.5 and 1.4 folds, respectively, compared to uninoculated controls. The repressor inactivated phosphate solubilizers better served the purpose of constitutive phosphate solubilization in pot experiments, where presence of other carbon sources (e.g., succinate) might repress mineral phosphate solubilization phenotype of wildtype strains.

  4. Derepression of Mineral Phosphate Solubilization Phenotype by Insertional Inactivation of iclR in Klebsiella pneumoniae

    PubMed Central

    Pandya, Maharshi; Jog, Rahul; G, Naresh Kumar; Rajkumar, Shalini

    2015-01-01

    The mode of succinate mediated repression of mineral phosphate solubilization and the role of repressor in suppressing phosphate solubilization phenotype of two free-living nitrogen fixing Klebsiella pneumoniae strains was studied. Organic acid mediated mineral phosphate solubilization phenotype of oxalic acid producing Klebsiella pneumoniae SM6 and SM11 were transcriptionally repressed by IclR in presence of succinate as carbon source. Oxalic acid production and expression of genes of the glyoxylate shunt (aceBAK) was found only in glucose but not in succinate- and glucose+succinate-grown cells. IclR, repressor of aceBAK operon, was inactivated using an allelic exchange system resulting in derepressed mineral phosphate solubilization phenotype through constitutive expression of the glyoxylate shunt. Insertional inactivation of iclR resulted in increased activity of the glyoxylate shunt enzymes even in succinate-grown cells. An augmented phosphate solubilization up to 54 and 59% soluble phosphate release was attained in glucose+succinate-grown SM6Δ and SM11Δ strains respectively, compared to glucose-grown cells, whereas phosphate solubilization was absent or negligible in wildtype cells grown in glucose+succinate. Both wildtype and iclR deletion strains showed similar indole-3-acetic acid production. Wheat seeds inoculated with wildtype SM6 and SM11 improved both root and shoot length by 1.2 fold. However, iclR deletion SM6Δ and SM11Δ strains increased root and shoot length by 1.5 and 1.4 folds, respectively, compared to uninoculated controls. The repressor inactivated phosphate solubilizers better served the purpose of constitutive phosphate solubilization in pot experiments, where presence of other carbon sources (e.g., succinate) might repress mineral phosphate solubilization phenotype of wildtype strains. PMID:26381651

  5. [Interaction between Bifidobacterium bifidum, Proteus vulgaris, and Klebsiella