Multicenter Evaluation of a New Shortened Peptide Nucleic Acid Fluorescence In Situ Hybridization Procedure for Species Identification of Select Gram-Negative Bacilli from Blood Cultures▿

PubMed Central

Morgan, Margie; Marlowe, Elizabeth; Della-Latta, Phyllis; Salimnia, Hossein; Novak-Weekley, Susan; Wu, Fann; Crystal, Benjamin S.

2010-01-01

A shortened protocol for two peptide nucleic acid fluorescence in situ hybridization (PNA FISH) assays for the detection of Gram-negative bacilli from positive blood cultures was evaluated in a multicenter trial. There was 100% concordance between the two protocols for each assay (368 of 368 and 370 of 370 results) and 99.7% (367 of 368 and 369 of 370 results) agreement with routine laboratory techniques. PMID:20357212

Comparative effects of single-dose ceftriaxone versus three oral antibiotic regimens on stool colonization by resistant bacilli in children.

PubMed

Toltzis, Philip; Dul, Michael; O'Riordan, Mary Ann; Toltzis, Hasida; Blumer, Jeffrey L

2007-01-01

The use of short-term intramuscular ceftriaxone for pediatric ambulatory conditions raises concerns regarding the promotion of resistance among colonizing enteric bacteria. This study was designed to assess the prevalence of stool colonization with resistant Gram-negative bacilli after single-dose ceftriaxone treatment compared with other regimens for acute otitis media. Children age 3 months to 7 years and diagnosed with acute otitis media were randomized to receive treatment with single-dose ceftriaxone or with oral cefprozil, amoxicillin or azithromycin. Stool samples were obtained at enrollment and then 3-5 days, 10-14 days, and 28-30 days after therapy was initiated and screened for the presence of facultative Gram-negative bacilli resistant to ceftriaxone, cefprozil, amoxicillin, piperacillin, piperacillin-tazobactam and tobramycin. Mean prevalence of colonization by resistant organisms for each treatment group was compared at each time point. One thousand nine subjects were enrolled. The prevalence of colonization by a Gram-negative bacillus resistant to at least 1 of the screening antibiotics decreased after receipt of ceftriaxone but returned close to values measured at study entry by 30 days. A qualitatively similar pattern was noted for the 3 other regimens, but a quantitatively greater decrease in the prevalence of colonization by a resistant bacterium was noted at the 3- to 5-day and 10- to 14-day visits among azithromycin recipients (P < 0.001). Colonization by a Gram-negative bacillus resistant specifically to ceftriaxone was unusual at each study visit, regardless of treatment assignment. A single intramuscular dose of ceftriaxone had a similar effect on the prevalence of antibiotic-resistant Gram-negative facultative bacilli in the stool of healthy children when compared with commonly used oral agents.

Comparison of Bruker Biotyper Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometer to BD Phoenix Automated Microbiology System for Identification of Gram-Negative Bacilli▿

PubMed Central

Saffert, Ryan T.; Cunningham, Scott A.; Ihde, Sherry M.; Monson Jobe, Kristine E.; Mandrekar, Jayawant; Patel, Robin

2011-01-01

We compared the BD Phoenix automated microbiology system to the Bruker Biotyper (version 2.0) matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry (MS) system for identification of Gram-negative bacilli, using biochemical testing and/or genetic sequencing to resolve discordant results. The BD Phoenix correctly identified 363 (83%) and 330 (75%) isolates to the genus and species level, respectively. The Bruker Biotyper correctly identified 408 (93%) and 360 (82%) isolates to the genus and species level, respectively. The 440 isolates were grouped into common (308) and infrequent (132) isolates in the clinical laboratory. For the 308 common isolates, the BD Phoenix and Bruker Biotyper correctly identified 294 (95%) and 296 (96%) of the isolates to the genus level, respectively. For species identification, the BD Phoenix and Bruker Biotyper correctly identified 93% of the common isolates (285 and 286, respectively). In contrast, for the 132 infrequent isolates, the Bruker Biotyper correctly identified 112 (85%) and 74 (56%) isolates to the genus and species level, respectively, compared to the BD Phoenix, which identified only 69 (52%) and 45 (34%) isolates to the genus and species level, respectively. Statistically, the Bruker Biotyper overall outperformed the BD Phoenix for identification of Gram-negative bacilli to the genus (P < 0.0001) and species (P = 0.0005) level in this sample set. When isolates were categorized as common or infrequent isolates, there was statistically no difference between the instruments for identification of common Gram-negative bacilli (P > 0.05). However, the Bruker Biotyper outperformed the BD Phoenix for identification of infrequently isolated Gram-negative bacilli (P < 0.0001). PMID:21209160

Evaluation of the in vitro activity of levornidazole, its metabolites and comparators against clinical anaerobic bacteria.

PubMed

Hu, Jiali; Zhang, Jing; Wu, Shi; Zhu, Demei; Huang, Haihui; Chen, Yuancheng; Yang, Yang; Zhang, Yingyuan

2014-12-01

This study evaluated the in vitro anti-anaerobic activity and spectrum of levornidazole, its metabolites and comparators against 375 clinical isolates of anaerobic bacteria, including Gram-negative bacilli (181 strains), Gram-negative cocci (11 strains), Gram-positive bacilli (139 strains) and Gram-positive cocci (44 strains), covering 34 species. Minimum inhibitory concentrations (MICs) of levornidazole, its five metabolites and three comparators against these anaerobic isolates were determined by the agar dilution method. Minimum bactericidal concentrations (MBCs) of levornidazole and metronidazole were measured against 22 strains of Bacteroides fragilis. Levornidazole showed good activity against B. fragilis, other Bacteroides spp., Clostridium difficile, Clostridium perfringens and Peptostreptococcus magnus, evidenced by MIC90 values of 0.5, 1, 0.25, 2 and 1mg/L, respectively. The activity of levornidazole and the comparators was poor for Veillonella spp. Generally, levornidazole displayed activity similar to or slightly higher than that of metronidazole, ornidazole and dextrornidazole against anaerobic Gram-negative bacilli, Gram-positive bacilli and Gram-positive cocci, especially B. fragilis. Favourable anti-anaerobic activity was also seen with levornidazole metabolites M1 and M4 but not M2, M3 or M5. For the 22 clinical B. fragilis strains, MBC50 and MBC90 values of levornidazole were 2mg/L and 4mg/L, respectively. Both MBC50/MIC50 and MBC90/MIC90 ratios of levornidazole were 4, similar to those of metronidazole. Levornidazole is an important anti-anaerobic option in clinical settings in terms of its potent and broad-spectrum in vitro activity, bactericidal property, and the anti-anaerobic activity of its metabolites M1 and M4. Copyright © 2014 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

Long-term outcome of acute prosthetic joint infections due to gram-negative bacilli treated with retention of prosthesis.

PubMed

Jaén, N; Martínez-Pastor, J C; Muñoz-Mahamud, E; García-Ramiro, S; Bosch, J; Mensa, J; Soriano, A

2012-09-01

To update the clinical information of the 47 patients with a prosthetic joint infection due to Gram-negative bacilli included in a previous study and to reassess the predictors of failure after a longer follow-up. Using the electronic files of our hospital, all the information regarding readmissions to the hospital, new surgical procedures and the reason for the new surgery (infection, aseptic loosening), and the last visit in the hospital were registered. The medical chart of the 35 patients that were considered in remission in the previous publication was reviewed. In 30 patients no clinical evidence of failure was detected and no additional surgery on the previously infected prosthesis was necessary and they were considered in long-term remission. In 5 cases a late complication was identified. One case had a reinfection due to coagulase-negative staphylococci after 22 months from the open debridement and required a 2-stage revision surgery. The other 4 cases developed an aseptic loosening and it was necessary to perform a 1-stage exchange. Receiving a fluoroquinolone when all the Gram-negatives involved in the infection were susceptible to fluoroquinolones was the only factor associated with remission in the univariate analysis (p=0.002). After a long-term follow-up, our results support the importance of using fluoroquinolones in acute PJI due to Gram-negative bacilli.

Microbiology of Drycleaning

PubMed Central

Banville, Robert R.; McNeil, Ethel

1966-01-01

An appreciable number of bacteria on contaminated fabric survived modern drycleaning procedures. Various stages in the process, especially steam pressing, reduced the total number of bacteria, but viable organisms were found on certain areas of garments even after pressing. A significant number of bacteria were redeposited on clean fabric during the washing of ordinary soiled garments in drycleaning units. These bacteria included gram-positive cocci, diphtheroid bacilli, and gram-positive sporeformers. Gram-negative bacilli were seldom found, although some gram-negative bacilli survived drycleaning. The redeposited organisms apparently came mainly from other garments in the same loads, as few bacteria were isolated from the filtered solvent used for washing. The number of bacteria in the drycleaning washwheel was highest shortly after the beginning of the wash, and decreased, with the exchange of solvent in the wheel, to a low level at the end. Although it appears that in most cases several factors combine to reduce to a low level the numbers of bacteria on articles cleaned in a well-operated drycleaning plant, it would seem that under certain conditions pathogenic microorganisms could be disseminated by drycleaning. Images Fig. 2 PMID:4958148

In Vitro Activities of Y-688, a New 7-Substituted Fluoroquinolone, against Anaerobic Bacteria

PubMed Central

MacGowan, A. P.; Bowker, K. E.; Wootton, M.; Holt, H. A.; Reeves, D. S.

1998-01-01

The in vitro activities of Y-688, a new 7-substituted fluoroquinolone derivative, against 317 nonduplicate anaerobic isolates were determined. Eighty-five percent of the Bacteroides fragilis group (n = 89) were inhibited by ≤2 mg of Y-688 per liter, while 78, 100, 89, and 98% of gram-negative bacilli (n = 135), gram-positive cocci (n = 59), and non-spore-forming (n = 58) and spore-forming (n = 51) gram-positive bacilli, respectively, were inhibited by ≤1 mg of Y-688 per liter. PMID:9527797

Use of elastin fibre detection in the diagnosis of ventilator associated pneumonia.

PubMed

el-Ebiary, M; Torres, A; González, J; Martos, A; Puig de la Bellacasa, J; Ferrer, M; Rodriguez-Roisin, R

1995-01-01

Elastin fibre detection could be a simple and reliable marker of ventilator associated pneumonia. To confirm this, a prospective study was undertaken to evaluate the diagnostic yield of elastin fibre detection in the diagnosis of ventilator associated pneumonia. Seventy eight mechanically ventilated patients were evaluated by examining endotracheal aspirates for the presence of elastin fibres. All patients were previously treated with antibiotics. Quantitative bacterial cultures of endotracheal aspirates and protected specimen brush samples were also performed. Patients were classified into three diagnostic categories: group 1, definite pneumonia (n = 25); group 2, probable pneumonia (n = 35); and group 3, controls (n = 18). Patients with definite and probable pneumonia were grouped together. The presence of elastin fibres in endotracheal aspirate samples was more frequent in groups 1 and 2, being found in 19 of the 60 patients compared with five of the control group. Although the presence of elastin fibres had a low sensitivity (32%), it was a reasonably specific marker (72%) of pneumonia. This specificity increased to 86% and 81% respectively when only Gram negative bacilli and Pseudomonas aeruginosa pneumonia were considered. Again, calculated sensitivity was 43% and 44% when analysing cases infected by Gram negative bacilli and Ps aeruginosa, respectively. The negative predictive value of the detection of elastin fibres in pneumonia caused by Ps aeruginosa was 81%. Detection was more frequent with infection by Gram negative bacilli (14/19), particularly with Ps aeruginosa (8/14). By contrast, pneumonia due to Gram positive cocci or non-bacterial agents uncommonly resulted in positive elastin fibre preparations (4/19, 21%). When analysing patients with and without chronic obstructive pulmonary disease, the diagnostic value of elastin fibre detection did not change. Potassium hydroxide preparation of elastin fibres is a rapid and simple specific marker of ventilator associated pneumonia and may be a useful technique to help diagnose pulmonary infections in mechanically ventilated patients, although this assessment is at present limited to patients without adult respiratory distress syndrome.

Use of elastin fibre detection in the diagnosis of ventilator associated pneumonia.

PubMed Central

el-Ebiary, M.; Torres, A.; González, J.; Martos, A.; Puig de la Bellacasa, J.; Ferrer, M.; Rodriguez-Roisin, R.

1995-01-01

BACKGROUND--Elastin fibre detection could be a simple and reliable marker of ventilator associated pneumonia. To confirm this, a prospective study was undertaken to evaluate the diagnostic yield of elastin fibre detection in the diagnosis of ventilator associated pneumonia. METHODS--Seventy eight mechanically ventilated patients were evaluated by examining endotracheal aspirates for the presence of elastin fibres. All patients were previously treated with antibiotics. Quantitative bacterial cultures of endotracheal aspirates and protected specimen brush samples were also performed. Patients were classified into three diagnostic categories: group 1, definite pneumonia (n = 25); group 2, probable pneumonia (n = 35); and group 3, controls (n = 18). RESULTS--Patients with definite and probable pneumonia were grouped together. The presence of elastin fibres in endotracheal aspirate samples was more frequent in groups 1 and 2, being found in 19 of the 60 patients compared with five of the control group. Although the presence of elastin fibres had a low sensitivity (32%), it was a reasonably specific marker (72%) of pneumonia. This specificity increased to 86% and 81% respectively when only Gram negative bacilli and Pseudomonas aeruginosa pneumonia were considered. Again, calculated sensitivity was 43% and 44% when analysing cases infected by Gram negative bacilli and Ps aeruginosa, respectively. The negative predictive value of the detection of elastin fibres in pneumonia caused by Ps aeruginosa was 81%. Detection was more frequent with infection by Gram negative bacilli (14/19), particularly with Ps aeruginosa (8/14). By contrast, pneumonia due to Gram positive cocci or non-bacterial agents uncommonly resulted in positive elastin fibre preparations (4/19, 21%). When analysing patients with and without chronic obstructive pulmonary disease, the diagnostic value of elastin fibre detection did not change. CONCLUSIONS--Potassium hydroxide preparation of elastin fibres is a rapid and simple specific marker of ventilator associated pneumonia and may be a useful technique to help diagnose pulmonary infections in mechanically ventilated patients, although this assessment is at present limited to patients without adult respiratory distress syndrome. PMID:7886642

[Ants as carriers of microorganisms in hospital environments].

PubMed

Pereira, Rogério Dos Santos; Ueno, Mariko

2008-01-01

Concern exists regarding the real possibility of public health threats caused by pathogenic agents that are carried by urban ants. The present study had the objective of isolating and identifying the microorganisms that are associated with ants in hospital environments. One hundred and twenty-five ants of the same species were collected from different units of a university hospital. Each ant was collected using a swab soaked with physiological solution and was transferred to a tube containing brain heart infusion broth and incubated at 35 degrees C for 24 hours. From each tube, with growth, inoculations were made into specific culturing media, to isolate any microorganisms. The ants presented a high capacity for carrying microorganism groups: spore-producing Gram-positive bacilli 63.5%, Gram-negative bacilli 6.3%, Gram-positive cocci 23.1%, filamentous fungi 6.7% and yeast 0.5%. Thus, it can be inferred that ants may be one of the agents responsible for disseminating microorganisms in hospital environments.

Emergence of Plasmid-Mediated Fosfomycin-Resistance Genes among Escherichia coli Isolates, France.

PubMed

Benzerara, Yahia; Gallah, Salah; Hommeril, Baptiste; Genel, Nathalie; Decré, Dominique; Rottman, Martin; Arlet, Guillaume

2017-09-01

FosA, a glutathione S-transferase that inactivates fosfomycin, has been reported as the cause of enzymatic resistance to fosfomycin. We show that multiple lineages of FosA-producing extended spectrum β-lactamase Escherichia coli have circulated in France since 2012, potentially reducing the efficacy of fosfomycin in treating infections with antimicrobial drug-resistant gram-negative bacilli.

Ticarcillin in Combination with Cephalothin or Gentamicin as Empiric Antibiotic Therapy in Granulocytopenic Cancer Patients

PubMed Central

Schimpff, Stephen C.; Landesman, Sheldon; Hahn, Davis M.; Standiford, Harold C.; Fortner, Clarence L.; Young, Viola Mae; Wiernik, Peter H.

1976-01-01

Ticarcillin was used in combination with either cephalothin or gentamicin as initial empiric antibiotic therapy for 127 patient trials of suspected infection in granulocytopenic cancer patients. Bacteremia was present in 20%, nonbacteremic microbiologically documented infections in 21%, clinically documented infections in 23%, and possible infections in 5%; infection was doubtful in 31%. Although Staphylococcus aureus was the most common single organism isolated (23%), gram-negative bacilli accounted for 54% of all pathogens. Both antibiotic regimens were highly efficacious, with complete resolution in 46% of bacteremias, 88% of nonbacteremic microbiologically documented infections, and 95% of clinically documented infections. Among bacteremias, 8 of 9 caused by S. aureus but only 4 of 15 (27%) caused by gram-negative bacilli were completely resolved with these antibiotic combinations. Reasons for nonresponse in bacteremias were persistent granulocytopenia, mixed infection and, in two patients, antibiotic-resistant organisms. Toxicities other than hypokalemia were minimal. Although the rate of further infections was high overall (18/127), only one occurred among the 39 patients with <4 days of antibiotic therapy. Ticarcillin in combination with either cephalothin or gentamicin was effective as initial empiric therapy of suspected infection in granulocytopenic cancer patients. PMID:795372

Appropriateness of gram-negative agent use at a tertiary care hospital in the setting of significant antimicrobial resistance.

PubMed

Vora, Neil M; Kubin, Christine J; Furuya, E Yoko

2015-01-01

Background.  Practicing antimicrobial stewardship in the setting of widespread antimicrobial resistance among gram-negative bacilli, particularly in urban areas, is challenging. Methods.  We conducted a retrospective cross-sectional study at a tertiary care hospital with an established antimicrobial stewardship program in New York, New York to determine appropriateness of use of gram-negative antimicrobials and to identify factors associated with suboptimal antimicrobial use. Adult inpatients who received gram-negative agents on 2 dates, 1 June 2010 or 1 December 2010, were identified through pharmacy records. Clinical data were collected for each patient. Use of gram-negative agents was deemed optimal or suboptimal through chart review and according to hospital guidelines. Data were compared using χ(2) or Fischer's exact test for categorical variables and Student t test or Mann-Whitney U test for continuous variables. Results.  A total of 356 patients were included who received 422 gram-negative agents. Administration was deemed suboptimal in 26% of instances, with the most common reason being spectrum of activity too broad. In multivariable analysis, being in an intensive care unit (adjusted odds ratio [aOR], .49; 95% confidence interval [CI], .29-.84), having an infectious diseases consultation within the previous 7 days (aOR, .52; 95% CI, .28-.98), and having a history of multidrug-resistant gram-negative bacilli within the past year (aOR, .24; 95% CI, .09-.65) were associated with optimal gram-negative agent use. Beta-lactam/beta-lactamase inhibitor combination drug use (aOR, 2.6; 95% CI, 1.35-5.16) was associated with suboptimal use. Conclusions.  Gram-negative agents were used too broadly despite numerous antimicrobial stewardship program activities.

Cerebral vasculitis and Cardiobacterium valvarum endocarditis.

PubMed

Abraham, R; Irwin, R B; Kannappan, D; Isalska, B; Koroma, M; Younis, N

2012-11-01

We present a case of aortic and tricuspid native valve endocarditis in which Cardiobacterium valvarum was isolated from the blood culture of a 65-year-old man. Cardiobacterium valvarum is a fastidious, Gram-negative bacillus. The genus Cardiobacterium encompasses two species - Cardiobacterium valvarum and Cardiobacterium hominis. Although both species rarely feature as the aetiological agent of endocarditis, Cardiobacterium hominis has a higher incidence than Cardiobacterium valvarum. For this causative organism, we believe this is the first report of fatality prior to surgical intervention and the first clinical course to be complicated by cerebral vasculitis. Native valve endocarditis caused by Gram-negative bacilli is extremely rare and identification of isolates may require the use of reference laboratories with molecular identification techniques.

A molecular gram stain using broad range PCR and pyrosequencing technology: a potentially useful tool for diagnosing orthopaedic infections.

PubMed

Kobayashi, Naomi; Bauer, Thomas W; Togawa, Daisuke; Lieberman, Isador H; Sakai, Hiroshige; Fujishiro, Takaaki; Tuohy, Marion J; Procop, Gary W

2005-06-01

The bacteria associated with orthopaedic infections are usually common gram-positive and gram-negative bacteria. This fundamental grouping of bacteria is a necessary first step in the selection of appropriate antibiotics. Since polymerase chain reaction (PCR) is more rapid and may be more sensitive than culture, we developed a postamplification pyrosequencing method to subcategorize bacteria based on a few nucleotide polymorphisms in the 16S rRNA gene. We validated this method using well-characterized strains of bacteria and applied it to specimens from spinal surgery cases with suspected infections. Lysates of 114 bacteria including 75 species were created following standard cultivation to obtain DNA. The DNA was amplified by a broad-range real-time PCR. The amplicons were evaluated by pyrosequencing and were classified as gram-positive, gram-negative, or acid-fast bacilli based on the first three to five nucleotides sequenced. In addition, clinical cases of suspected infection were obtained from spinal surgery. The results of the "molecular Gram stain" were compared with the results of traditional Gram stain and culture. The lysates of 107 (93.9%) of the bacteria extracts tested were appropriately categorized as gram-positive and gram-negative or as acid-fast bacilli on the basis of this assay. The sensitivity and specificity of this assay were 100% and 97.4% for gram-positive and 88.3% and 100% for gram-negative isolates. All of the five clinical samples were appropriately categorized as containing gram-positive or gram-negative bacteria with this assay. This study demonstrates that high sensitivity and specificity of a molecular gram stain may be achieved using broad-range real-time PCR and pyrosequencing.

Invasive bacterial infections in a pediatric oncology unit in a tertiary care center.

PubMed

Trehan, A; Totadri, S; Gautam, V; Bansal, D; Ray, P

2014-01-01

Multidrug resistant (MDR) pathogens are becoming a major problem worldwide, more so in the immunocompromised hosts resulting in the urgent need of antibiotic stewardship. To analyze the organisms isolated and the drug resistance pattern in a pediatric oncology unit. Data pertaining to infections with 128 positive cultures in patients with febrile neutropenia over a period of 1-year are presented. The unit antibiotic policy is decided depending on the sensitivity of the prevailing common organisms. We isolated Gram-negative organisms in 56% cases. Escherichia coli and Klebseilla were the most frequent lactose fermenting Gram-negative Bacilli and Pseudomonas and Acinetobacter the nonfermenting Gram-negative Bacilli. Only 20-30% of the Gram-negative organisms cultured were sensitive to a 3rd/4th generation cephalosporin. The combination of a beta-lactam/inhibitor covered 2/3rd of Gram-negative organisms. About 80% of the organisms were sensitive to carbapenems. There was no colistin resistance. About 44% of our cultures grew a Gram-positive bacterial organism and included coagulase negative Staphylococcus. We had an incidence of methicillin resistant Staphylococcus aureus to be 30%. About 30% of the enterococci isolated in our unit were vancomycin-resistant enterococci. About 23% of patients with a positive bacterial culture died. Infections in pediatric cancer patient's account for about 15-20% of the deaths in developing countries as these patients are at a high risk for developing MDR infections. Resistance rates among Gram-positive and Gram-negative organisms have increased worldwide. Every unit needs a rational antibiotic policy. Antibiotic de-escalation and judicious decrease in the duration of antibiotics needs to be practiced.

Clinical evaluation of the Vitek ANI card for identification of anaerobic bacteria.

PubMed Central

Schreckenberger, P C; Celig, D M; Janda, W M

1988-01-01

An evaluation of the Vitek Anaerobe Identification (ANI) card was performed with 341 bacterial isolates, including 313 clinical isolates and 28 stock strains of anaerobic microorganisms. Identifications obtained with the ANI card were compared with those determined by conventional methods. The card identified 73.2% of 149 anaerobic gram-negative bacilli, 63.6% of 44 Clostridium spp., 65.8% of 38 anaerobic nonsporeforming gram-positive bacilli, and 69.1% of 110 anaerobic cocci, with no further testing required. When genus-level identifications were included, 83.9% of the anaerobic gram-negative bacilli, 70.5% of Clostridium spp., 73.7% of the anaerobic nonsporeforming gram-positive bacilli, and 73.6% of the anaerobic cocci were identified. Nineteen isolates (5.6%) produced identifications of good confidence but marginal separation or questionable biotype, in which the correct identification was listed with one or two other possible choices and extra tests were required and suggested. A total of 28 (8.2%) were not identified and 29 isolates (8.5%) were misidentified by the ANI card. Among the commonly isolated clinically significant anaerobes, the ANI card identified 100% of 55 Bacteroides fragilis and 100% of 8 Clostridium perfringens. Use of supplemental tests and expansion of the data base to include additional strains of organisms that are difficult to separate even with conventional methods may improve the accuracy of the ANI card as a method for identification of anaerobic bacteria in the clinical laboratory. PMID:3343321

Antibiotic-Resistant Gram Negative Bacilli in Meals Delivered at a General Hospital, Italy

PubMed Central

Plano, Maria Rosa Anna; Di Noto, Anna Maria; Firenze, Alberto; Sciortino, Sonia; Mammina, Caterina

2009-01-01

This study aimed at detecting the presence of antibiotic-resistant Gram-negatives in samples of meals delivered at the University General Hospital of Palermo, Italy. Antibiotic resistant Gram negatives were isolated in July—September 2007 ffrom cold dishes and food contact surfaces and utensils. Bacterial strains were submitted to susceptibility test and subtyped by random amplification of polymorphic DNA (RAPD). Forty-six of 55 (83.6%) food samples and 14 of 17 (82.3%) environmental swabs were culture positive for Gram negative bacilli resistant to at least one group of antibacterial drugs. A total of 134 antibiotic resistant strains, 51 fermenters and 83 non-fermenters, were recovered. Fermenters and non-fermenters showed frequencies as high as 97.8% of resistance to two or more groups of antibiotics and non fermenters were 28.9% resistant to more than three groups. Molecular typing detected 34 different profiles among the fermenters and 68 among the non-fermenters. Antibiotic resistance was very common among both fermenters and non-fermenters. However, the wide heterogeneity of RAPD patterns seems to support a prominent role of cross-contamination rather than a clonal expansion of a few resistant isolates. A contribution of commensal Gram negatives colonizing foods to a common bacterial resistance pool should not been overlooked. PMID:19750189

Antibiotic-resistant gram negative bacilli in meals delivered at a general hospital, Italy.

PubMed

Plano, Maria Rosa Anna; Di Noto, Anna Maria; Firenze, Alberto; Sciortino, Sonia; Mammina, Caterina

2009-01-01

This study aimed at detecting the presence of antibiotic-resistant Gram-negatives in samples of meals delivered at the University General Hospital of Palermo, Italy. Antibiotic resistant Gram negatives were isolated in July-September 2007 ffrom cold dishes and food contact surfaces and utensils. Bacterial strains were submitted to susceptibility test and subtyped by random amplification of polymorphic DNA (RAPD). Forty-six of 55 (83.6%) food samples and 14 of 17 (82.3%) environmental swabs were culture positive for Gram negative bacilli resistant to at least one group of antibacterial drugs. A total of 134 antibiotic resistant strains, 51 fermenters and 83 non-fermenters, were recovered. Fermenters and non-fermenters showed frequencies as high as 97.8% of resistance to two or more groups of antibiotics and non fermenters were 28.9% resistant to more than three groups. Molecular typing detected 34 different profiles among the fermenters and 68 among the non-fermenters. Antibiotic resistance was very common among both fermenters and non-fermenters. However, the wide heterogeneity of RAPD patterns seems to support a prominent role of cross-contamination rather than a clonal expansion of a few resistant isolates. A contribution of commensal Gram negatives colonizing foods to a common bacterial resistance pool should not been overlooked.

Bacteriological Assessment of Pneumonia Caused by Gram-Negative Bacteria in Patients Hospitalized in Intensive Care Unit.

PubMed

Guzek, A; Korzeniewski, K; Tomaszewski, D; Rybicki, Z; Zwolińska, E

2017-01-01

The article presents the results of 11-year study (2005-2015) of Gram-negative bacteria responsible for pneumonia in 2033 mechanically ventilated patients hospitalized in Intensive Care Unit. Of 8796 biological samples, consisting mainly of bronchial aspirate (97.9 %), 2056 bacterial strains were isolated and subjected to identification. VITEK 2 was used to determine drug susceptibility (classified according to the EUCAST criteria). ESBL, MBL and KPC-producing strains were identified by means of phenotypic methods using appropriate discs. The findings were that the predominant bacteria responsible for infections consisted of Enterobacteriaceae (42.0 %), Acinetobacter baumannii (37.2 %), Pseudomonas aeruginosa (16.1 %), and Stenotrophomonas maltophila (4.7 %). We observed a rise in the number of bacteria causing pneumonia throughout the study period, especially in S. maltophila and Enterobacteriaceae ESBL (+). Gram-negative bacilli were 100 % susceptible to colistin, apart from naturally resistant strains such as Proteus mirabilis, Serratia marcescens, whereas Enterobacteriaceae ESBL (+) were susceptible to imipenem and meropenem. Acinetobacter baumannii strains exhibited the lowest drug susceptibility. In conclusion, we report an increase in the prevalence of pneumonia associated with Gram-negative bacteria in mechanically ventilated intensive care patients. Colistin remains the most effective drug against the majority of Gram-negative bacteria. Therapeutic problems are common in the course of treatment of Acinetobacter baumannii infections.

Impact of fluoroquinolone administration on the emergence of fluoroquinolone-resistant gram-negative bacilli from gastrointestinal flora.

PubMed

Richard, P; Delangle, M H; Raffi, F; Espaze, E; Richet, H

2001-01-01

We assessed the risk factors for acquisition of fluoroquinolone-resistant, gram-negative organisms in the gastrointestinal tract of hospitalized patients. We analyzed stool samples from 204 patients and recovered fluoroquinolone-resistant, gram-negative organisms from 63. Receipt of fluoroquinolone during the month preceding admission was the only risk factor identified, whereas female sex, duration of hospitalization, exposure to indwelling devices, admission from another hospital, and history of infection were risk factors for fecal colonization after day 4.

Antibiotic susceptibility patterns of different bacteria isolated from patients with ventilator associated pneumonia (vap).

PubMed

Alqurashi, Abdulrahman M

2005-09-01

Ventilator associated pneumonia (VAP) is a frequent complication of mechanical ventilation (MV) and it is a leading cause of death in MV patients. The development of VAP has been demonstrated as being due to aspiration of oropharyngeal secretion, ventilator tubing condensate, or gastric contents that are colonized with pathogenic microorganisms. The aim of the present study is to isolate and identify bacteria that cause VAP and to study antibiotic susceptibility. This study was carried out on 95 patients who fulfilled the diagnostic criteria for VAP. Quantitative cultures of endotracheal aspirates (EA) using a cut-off point of 10(6) cfu/ml was done. The microbiological results revealed that gram negative bacilli were the most common bacterial agents responsible for VAP and accounted for 78.8% of all the causative agents. The most common isolated organisms were Klebsiella pnemouniae (30.9 %) followed by Pseudomonas aeruginosa (22.5%), Staphylococcus aureus (21.2%), Eschericia coli (12.8 %), Proteus spp. (9.8%), and Citrobacter spp. (2.8%). Blood cultures were positive in 25.9% of patients with Klebsiella pnemouniae in about 33.3%. From this study, it can be concluded that VAP is an important nosocomial infection. EA is a simple procedure to obtain respiratory samples and perform sensitivity testing in patients with VAP. Also, the commonest cause of VAP is gram negative bacilli.

Effectiveness of hand washing and disinfection methods in removing transient bacteria after patient nursing.

PubMed Central

Ojajärvi, J.

1980-01-01

The effectiveness of various hand washing and disinfection methods in removing transient skin bacteria was studied in hospital after dry or moist contamination of the hands when nursing burn patients. The results were compared with those of laboratory tests with volunteers. A fairly good correlation of the bacterial reductions existed between hospital and laboratory tests. All other methods removed Staph. aureus from the hands more effectively than liquid soap. Gram-negative bacilli were more easily removed than staphylococci, even with soap wash alone. In hospital, none of the washing and disinfection methods always removed all patient-borne bacteria from the hands. After dry or moist contamination and subsequent washing with soap only, colonies of Staph. aureus were often detected in finger-print samples. Staphylococci were more often completely removed by a 4% chlorhexidine detergent scrub and alcoholic solutions (either with or without previous soap wash) than by liquid soap, hexachlorophene or iodophor preparations. Gram-negative bacilli were more easily removed by all the washing and disinfection methods. After moist contamination, Gram-negative bacilli were more often completely removed from the hands by ethanol than by other treatments. The results of the present study emphasize the importance of always using gloves when nursing a profuse spreader of bacteria or one who must be protected from infection. PMID:6778914

[Periodontal microbiota and microorganisms isolated from heart valves in patients undergoing valve replacement surgery in a clinic in Cali, Colombia].

PubMed

Moreno, Sandra; Parra, Beatriz; Botero, Javier E; Moreno, Freddy; Vásquez, Daniel; Fernández, Hugo; Alba, Sandra; Gallego, Sara; Castillo, Gilberto; Contreras, Adolfo

2017-12-01

Periodontitis is an infectious disease that affects the support tissue of the teeth and it is associated with different systemic diseases, including cardiovascular disease. Microbiological studies facilitate the detection of microorganisms from subgingival and cardiovascular samples. To describe the cultivable periodontal microbiota and the presence of microorganisms in heart valves from patients undergoing valve replacement surgery in a clinic in Cali. We analyzed 30 subgingival and valvular tissue samples by means of two-phase culture medium, supplemented blood agar and trypticase soy agar with antibiotics. Conventional PCR was performed on samples of valve tissue. The periodontal pathogens isolated from periodontal pockets were: Fusobacterium nucleatum (50%), Prevotella intermedia/ nigrescens (40%), Campylobacter rectus (40%), Eikenella corrodens (36.7%), Gram negative enteric bacilli (36.7%), Porphyromonas gingivalis (33.3%), and Eubacterium spp. (33.3%). The pathogens isolated from the aortic valve were Propionibacterium acnes (12%), Gram negative enteric bacilli (8%), Bacteroides merdae (4%), and Clostridium bifermentans (4%), and from the mitral valve we isolated P. acnes and Clostridium beijerinckii. Conventional PCR did not return positive results for oral pathogens and bacterial DNA was detected only in two samples. Periodontal microbiota of patients undergoing surgery for heart valve replacement consisted of species of Gram-negative bacteria that have been associated with infections in extraoral tissues. However, there is no evidence of the presence of periodontal pathogens in valve tissue, because even though there were valve and subgingival samples positive for Gram-negative enteric bacilli, it is not possible to maintain they corresponded to the same phylogenetic origin.

Isolation and determination of four potential antimicrobial components from Pseudomonas aeruginosa extracts

PubMed Central

Xu, Ling-Qing; Zeng, Jian-Wen; Jiang, Chong-He; Wang, Huan; Li, Yu-Zhen; Wen, Wei-Hong; Li, Jie-Hua; Wang, Feng; Ting, Wei-Jen; Sun, Zi-Yong; Huang, Chih-Yang

2017-01-01

Background: Pseudomonas aeruginosa can cause disease and also can be isolated from the skin of healthy people. Additionally, it exhibits certain antimicrobial effects against other microorganisms. Methods: We collected 60 strains of P. aeruginosa and screened their antimicrobial effects against Staphylococcus aureus (ATCC 25923) using the filter paper-disk method, the cross-streaking method and the co-culture method and then evaluated the antimicrobial activity of the chloroform-isolated S. aureus extracts against methicillin-resistant S. aureus (MRSA, Gram-positive cocci), vancomycin intermediate-resistant S. aureus (VISA, Gram-positive cocci), Corynebacterium spp. (CS, Gram-positive bacilli), Acinetobacter baumannii (AB, Gram-negative bacilli), Moraxella catarrhalis (MC, Gram-negative diplococcus), Candida albicans (CA, fungi), Candida tropicalis (CT, fungi), Candida glabrata (CG, fungi) and Candida parapsilosis (CP, fungi). Results: The PA06 and PA46 strains have strong antimicrobial effects. High-performance liquid chromatography (HPLC) analysis revealed that the major components of PA06 and PA46 that exhibit antimicrobial activity are functionally similar to phenazine-1-carboxylic acid (PCA) and pyocyanin. Preparative HPLC was performed to separate and isolate the 4 major potential antimicrobial components: PA06ER10, PA06ER16, PA06ER23 and PA06ER31. Further, the molecular masses of PA06ER10 (260.1), PA06ER16 (274.1), PA06ER23 (286.1) and PA06ER31 (318.2) were determined by electrospray ionization (ESI) mass spectrometry. Conclusion: P. aeruginosa can produce small molecules with potential antimicrobial activities against MRSA, VISA, CS, MC, CA, CT, CG and CP but not against AB. PMID:29200950

Quantifying Glosair™ 400 efficacy for surface disinfection of American Type Culture Collection strains and micro-organisms recently isolated from intensive care unit patients.

PubMed

Herruzo, R; Vizcaíno, M J; Herruzo, I

2014-07-01

Microbial contamination of hospital surfaces may be a source of infection for hospitalized patients. We evaluated the efficacy of Glosair™ 400 against two American Type Culture Collection strains and 18 clinical isolates, placed on glass germ-carriers. Carriers were left to air-dry for 60 min and then exposed to a cycle before detection of any surviving micro-organisms. Antibiotic-susceptible Gram-negative bacilli were less susceptible (although not significantly) to this technique than resistant Gram-negative bacilli or Gram-positive cocci and yeasts (3, 3.4 and 4.6 log10 reduction, respectively). In conclusion, in areas that had not been cleaned, aerosolized hydrogen peroxide obtained >3 log10 mean destruction of patients' micro-organisms. Copyright © 2014 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

[Anaerobic Gram-negative bacilli involved in the etiopathogeny of the abscesses of superficial fascial spaces of the face and neck].

PubMed

Băncescu, Gabriela; Băncescu, A; Dumitriu, Silvia; Skaug, N

2008-01-01

The aim of this study was to isolate and identify at species level the strains of anaerobic Gram-negative bacilli (GNB) from pus samples collected in patients with abscesses of fascial spaces of the face and neck. Microscopy of Gram-stained smears and cultures were performed in each specimen. The strictly anaerobic GNB strains were identified using the conventional methods of diagnosis and the Rapid ID 32 A system. In addition, the other strains isolated in association with these bacteria were identified at least to genus level. The 28 anaerobic GNB isolates belonged to: Fusobacterium nucleatum and different species of Prevotella (4 species) and Bacteroides (3 species). The anaerobic GBN strains were recovered--either alone or in association with other migroorganisms--in more than half of all investigated samples and represented about 40% of all isolates. The most frequently isolated species were P> melaninogenica and B. ureolyticus.

Antibacterial properties of propolis (bee glue).

PubMed Central

Grange, J M; Davey, R W

1990-01-01

Propolis (bee glue) was found to have antibacterial activity against a range of commonly encountered cocci and Gram-positive rods, including the human tubercle bacillus, but only limited activity against Gram-negative bacilli. These findings confirm previous reports of antimicrobial properties of this material, possibly attributable to its high flavonoid content. PMID:2182860

Hospitalized tornado victims.

PubMed

Harris, L F

1992-01-01

We reviewed the clinical findings of 59 patients admitted to Huntsville's three hospitals during the first 24 hours after the November 15, 1989, tornado and as a result of injury caused by the tornado. Fracture of a bone was the most common injury followed by soft tissue trauma and infection. A variety of non-traumatic conditions also were encountered. Fractures were more frequent above the waist than below and five fractures became infected resulting in osteomyelitis. Infections most often involved the urinary tract and bone and were caused primarily by aerobic gram-negative bacilli. The hospital mortality rate was 7%.

10 x '20 Progress--development of new drugs active against gram-negative bacilli: an update from the Infectious Diseases Society of America.

PubMed

Boucher, Helen W; Talbot, George H; Benjamin, Daniel K; Bradley, John; Guidos, Robert J; Jones, Ronald N; Murray, Barbara E; Bonomo, Robert A; Gilbert, David

2013-06-01

Infections caused by antibiotic-resistant bacteria, especially the "ESKAPE" pathogens, continue to increase in frequency and cause significant morbidity and mortality. New antimicrobial agents are greatly needed to treat infections caused by gram-negative bacilli (GNB) resistant to currently available agents. The Infectious Diseases Society of America (IDSA) continues to propose legislative, regulatory, and funding solutions to this continuing crisis. The current report updates the status of development and approval of systemic antibiotics in the United States as of early 2013. Only 2 new antibiotics have been approved since IDSA's 2009 pipeline status report, and the number of new antibiotics annually approved for marketing in the United States continues to decline. We identified 7 drugs in clinical development for treatment of infections caused by resistant GNB. None of these agents was included in our 2009 list of antibacterial compounds in phase 2 or later development, but unfortunately none addresses the entire spectrum of clinically relevant GNB resistance. Our survey demonstrates some progress in development of new antibacterial drugs that target infections caused by resistant GNB, but progress remains alarmingly elusive. IDSA stresses our conviction that the antibiotic pipeline problem can be solved by the collaboration of global leaders to develop creative incentives that will stimulate new antibacterial research and development. Our aim is the creation of a sustainable global antibacterial drug research and development enterprise with the power in the short term to develop 10 new, safe, and efficacious systemically administered antibiotics by 2020 as called for in IDSA's "10 × '20 Initiative."

The usefulness of changing focus during examination using Gram staining as initial diagnostic clue for infective tuberculosis.

PubMed

Atsukawa, Yoshiko; Kawakami, Sayoko; Asahara, Miwa; Ishigaki, Shinobu; Tanaka, Takashi; Ono, Yasuo; Nishiya, Hajime; Fujisaki, Ryuichi; Koga, Ichiro; Ota, Yasuo; Miyazawa, Yukihisa

2011-08-01

Gram staining is a useful technique for detecting bacteria but is highly questionable in detecting Mycobacterium tuberculosis. Its detection generally requires special staining, such as Ziehl-Neelsen staining. We experienced three cases in which tuberculosis was first suggested by Gram staining of sputum or pus, confirmed by Ziehl-Neelsen staining, and diagnosed by polymerase chain reaction or culture. To find colorless tubercle bacilli in clinical samples with various organisms, varying the focus to slightly longer and shorter during study of the slides is indispensable. We present criteria for detecting infective pulmonary tuberculosis in Gram staining. First, in the ordinary focus, weakly stained, thin, gram-positive bacilli are found; second, with a slightly longer focus distance, the thin, cord-like, conspicuous gram-positive bacilli can be observed; and third, with a shorter focus distance, the gram-positive bacilli have changed into the brightened, colorless, or ghost ones. Four laboratory technologists each evaluated 20 Gram-stained samples after being lectured on the criteria, with no prior information about the sample. They accurately evaluated the presence of the bacilli in Gram-stained preparations in more than 90% of samples containing 3+ bacilli on Ziehl-Neelsen staining. Gram staining is available as an easy and rapid initial clue to recognize highly infective tuberculosis.

Enteric gram-negative bacilli bloodstream infections: 17 years' experience in a neonatal intensive care unit.

PubMed

Cordero, Leandro; Rau, Rachel; Taylor, David; Ayers, Leona W

2004-06-01

To assess the occurrence of enteric gram-negative bacilli (EGNB) bloodstream infections (BSI) in a neonatal intensive care setting during a 17-year period in which a consistent antibiotic treatment program was in place. To document infections, outbreaks, or epidemics, emergence of antibiotic resistance, clinical correlates, and outcomes of the most prevalent EGNB (Escherichia coli, Klebsiella pneumoniae, and Enterobacter cloacae). This study analyzed demographic, clinical, and bacteriologic information from 360 infants born 1986-2002 who developed 633 blood culture-proven BSI. A total of 121 EGNB were isolated (E coli, K pneumoniae, and E cloacae). Early-onset BSI were discovered within 48 hours from birth, and late-onset BSI were those that occurred thereafter. Suspected early-onset BSI were treated with ampicillin and gentamicin, suspected late-onset BSI with vancomycin and gentamicin. Antibiotics were changed on the basis of organism antimicrobial susceptibility. Early-onset BSI were noted in 52 of 21,336 (244/100,000) live births (1986-1991), 40 of 20,402 (196/100,000) live births (1992-1997), and 25 of 17,926 (139/100,000) live births (1998-2002). Of these cases, 39 were caused by E coli and 4 by K pneumoniae. Antibiograms for E coli isolated during the last 5 years of the study showed an increase in antibiotic resistance that coincided with obstetric group B streptococcus antepartum antibiotic prophylaxis. Group B streptococcus declined from 41 to 4 cases from the first to the last period. Late-onset BSI increased from 111 to 230 cases from the first to the second 6-year study period and declined modestly (171 cases) during the last. Fifteen percent (78 cases) of late-onset BSI were caused by EGNB, 5% by other gram-negative bacilli, 67% primarily by coagulase-negative staphylococcus, and 13% by fungus. Nonspecific clinical and hematologic signs of late-onset BSI were similar across EGNB species, but necrotizing enterocolitis was often associated with E coli, whereas pneumonia and prolonged thrombocytopenia characterized K pneumoniae infections. No outbreaks or epidemics were observed, and strains of EGNB with evidence of extended spectrum beta-lactamase production were never isolated. Antepartum antibiotic prophylaxis may have increased antibiotic resistance in E coli isolates from early-onset BSI but has dramatically decreased group B streptococcus infections. Late-onset BSI caused by EGNB increased, but without changes in antibiotic susceptibility. In spite of medical advances, E coli, K pneumoniae, and E cloacae remain responsible for significant morbidity and mortality, especially in very low birth weight infants.

Gentamicin-loaded borate bioactive glass eradicates osteomyelitis due to Escherichia coli in a rabbit model.

PubMed

Xie, Zongping; Cui, Xu; Zhao, Cunju; Huang, Wenhai; Wang, Jianqiang; Zhang, Changqing

2013-07-01

The treatment of osteomyelitis induced by Gram-negative bacilli is rarely reported in the literature. This study established a rabbit tibia model of osteomyelitis induced by the Gram-negative bacillus Escherichia coli. Using this model, pellets composed of a chitosan-bonded mixture of borate bioactive glass and gentamicin were evaluated in vitro and in vivo for the treatment of osteomyelitis induced by Escherichia coli. Our results showed that the pellets in phosphate-buffered saline released gentamicin continuously over 26 days. Without the simultaneous use of a systemic antibiotic, the implantation of the gentamicin-loaded pellets into the osteomyelitis region of the tibia resulted in the eradication of 81.82% of infections, as determined by microbiological, histological and radiographic evaluation, and supported the ingrowth of new bone into the tibia defects after 6 weeks of implantation. The results indicate that the gentamicin-loaded borate bioactive glass implant, combining sustained drug release with the ability to support new bone formation, could provide a method for treating osteomyelitis induced by Gram-negative bacilli.

Gentamicin-Loaded Borate Bioactive Glass Eradicates Osteomyelitis Due to Escherichia coli in a Rabbit Model

PubMed Central

Xie, Zongping; Cui, Xu; Zhao, Cunju; Huang, Wenhai; Wang, Jianqiang

2013-01-01

The treatment of osteomyelitis induced by Gram-negative bacilli is rarely reported in the literature. This study established a rabbit tibia model of osteomyelitis induced by the Gram-negative bacillus Escherichia coli. Using this model, pellets composed of a chitosan-bonded mixture of borate bioactive glass and gentamicin were evaluated in vitro and in vivo for the treatment of osteomyelitis induced by Escherichia coli. Our results showed that the pellets in phosphate-buffered saline released gentamicin continuously over 26 days. Without the simultaneous use of a systemic antibiotic, the implantation of the gentamicin-loaded pellets into the osteomyelitis region of the tibia resulted in the eradication of 81.82% of infections, as determined by microbiological, histological and radiographic evaluation, and supported the ingrowth of new bone into the tibia defects after 6 weeks of implantation. The results indicate that the gentamicin-loaded borate bioactive glass implant, combining sustained drug release with the ability to support new bone formation, could provide a method for treating osteomyelitis induced by Gram-negative bacilli. PMID:23629702

Necrotizing suppurative nephritis in a Japanese black feedlot steer due to Proteus mirabilis infection.

PubMed

Abe, Tadatsugu; Iizuka, Ayako; Kojima, Hirokazu; Kimura, Kumiko; Shibahara, Tomoyuki; Haritani, Makoto

2017-04-05

A Japanese black feedlot steer suddenly died after exhibiting astasia and cramping of the extremities. Necropsy of the animal revealed that the right kidney was enlarged and pale with severe nephrolithiasis. The urinary bladder displayed mucosal hemorrhage. Upon bacteriological investigation, Proteus mirabilis was isolated from the liver, spleen, right kidney, lungs and urine. Histopathological examination revealed necrotizing suppurative nephritis with the presence of numerous gram-negative bacilli and fibrinous suppurative cystitis with no bacilli. Immunohistochemical analysis revealed that the bacteria and cytoplasm of the macrophages stained positively with P. mirabilis antiserum. Electron microscopy revealed the presence of numerous bacteria in the renal tubules. To our knowledge, this is the first report describing the histopathological aspects of nephritis caused by P. mirabilis in cattle.

10 × '20 Progress—Development of New Drugs Active Against Gram-Negative Bacilli: An Update From the Infectious Diseases Society of America

PubMed Central

Boucher, Helen W.; Talbot, George H.; Benjamin, Daniel K.; Bradley, John; Guidos, Robert J.; Jones, Ronald N.; Murray, Barbara E.; Bonomo, Robert A.; Gilbert, David

2013-01-01

Infections caused by antibiotic-resistant bacteria, especially the “ESKAPE” pathogens, continue to increase in frequency and cause significant morbidity and mortality. New antimicrobial agents are greatly needed to treat infections caused by gram-negative bacilli (GNB) resistant to currently available agents. The Infectious Diseases Society of America (IDSA) continues to propose legislative, regulatory, and funding solutions to this continuing crisis. The current report updates the status of development and approval of systemic antibiotics in the United States as of early 2013. Only 2 new antibiotics have been approved since IDSA's 2009 pipeline status report, and the number of new antibiotics annually approved for marketing in the United States continues to decline. We identified 7 drugs in clinical development for treatment of infections caused by resistant GNB. None of these agents was included in our 2009 list of antibacterial compounds in phase 2 or later development, but unfortunately none addresses the entire spectrum of clinically relevant GNB resistance. Our survey demonstrates some progress in development of new antibacterial drugs that target infections caused by resistant GNB, but progress remains alarmingly elusive. IDSA stresses our conviction that the antibiotic pipeline problem can be solved by the collaboration of global leaders to develop creative incentives that will stimulate new antibacterial research and development. Our aim is the creation of a sustainable global antibacterial drug research and development enterprise with the power in the short term to develop 10 new, safe, and efficacious systemically administered antibiotics by 2020 as called for in IDSA's “10 × '20 Initiative.” PMID:23599308

Fasting hypochlorhydria with gram positive gastric flora is highly prevalent in healthy old people.

PubMed Central

Husebye, E; Skar, V; Høverstad, T; Melby, K

1992-01-01

Fifteen healthy old people mean age 84 years (range 80-91 years), were examined to assess the effect of advanced age on the microecology of the upper gastrointestinal tract. Twelve of 15 (80%) were hypochlorhydric with pH 6.6 (0.3) (mean (SEM) and a mean bacterial count of 10(8) colony forming units (CFU) per ml (range 10(5)-10(10)) in fasting gastric aspirate. Normochlorhydric subjects had low counts (< or = 10(1) CFU/ml). The microbial flora was dominated by viridans streptococci, coagulase negative staphylococci, and Haemophilus sp. Only one subject harboured significant concentrations of Gram negative bacilli with Escherichia coli (10(4-5) CFU/ml) and Klebsiella (10(4-5)). Strict anaerobes were not found. The total concentration of short chain fatty acids in gastric aspirate was 10.6 (2.9) mmol/l (mean (SEM). Absence of significant, intraluminal fermentation of xylose to CO2 was shown by the 14C-d Xylose breath test, and ambulatory manometry showed preserved fasting motility pattern of the small intestine. Serum immunoglobulins were normal. Advanced age is accompanied by fasting hypochlorhydria and colonisation with mainly Gram positive flora in the upper gut. Other factors than old age and fasting hypochlorhydria are required for colonisation with Gram negative bacilli. PMID:1446855

Utility of Gram Staining for Evaluation of the Quality of Cystic Fibrosis Sputum Samples

PubMed Central

Nair, Bindu; Stapp, Jenny; Stapp, Lynn; Bugni, Linda; Van Dalfsen, Jill; Burns, Jane L.

2002-01-01

The microscopic examination of Gram-stained sputum specimens is very helpful in the evaluation of patients with community-acquired pneumonia and has also been recommended for use in cystic fibrosis (CF) patients. This study was undertaken to evaluate that recommendation. One hundred one sputum samples from CF patients were cultured for gram-negative bacilli and examined by Gram staining for both sputum adequacy (using the quality [Q] score) and bacterial morphology. Subjective evaluation of adequacy was also performed and categorized. Based on Q score evaluation, 41% of the samples would have been rejected despite a subjective appearance of purulence. Only three of these rejected samples were culture negative for gram-negative CF pathogens. Correlation between culture results and quantitative Gram stain examination was also poor. These data suggest that subjective evaluation combined with comprehensive bacteriology is superior to Gram staining in identifying pathogens in CF sputum. PMID:12149331

Utility of gram staining for evaluation of the quality of cystic fibrosis sputum samples.

PubMed

Nair, Bindu; Stapp, Jenny; Stapp, Lynn; Bugni, Linda; Van Dalfsen, Jill; Burns, Jane L

2002-08-01

The microscopic examination of Gram-stained sputum specimens is very helpful in the evaluation of patients with community-acquired pneumonia and has also been recommended for use in cystic fibrosis (CF) patients. This study was undertaken to evaluate that recommendation. One hundred one sputum samples from CF patients were cultured for gram-negative bacilli and examined by Gram staining for both sputum adequacy (using the quality [Q] score) and bacterial morphology. Subjective evaluation of adequacy was also performed and categorized. Based on Q score evaluation, 41% of the samples would have been rejected despite a subjective appearance of purulence. Only three of these rejected samples were culture negative for gram-negative CF pathogens. Correlation between culture results and quantitative Gram stain examination was also poor. These data suggest that subjective evaluation combined with comprehensive bacteriology is superior to Gram staining in identifying pathogens in CF sputum.

Case of a lung mass due to melioidosis in Mexico.

PubMed

Truong, Kimberly K; Moghaddam, Samer; Al Saghbini, Samer; Saatian, Bahman

2015-05-06

Melioidosis, an infection caused by the gram-negative bacterium Burkholderia pseudomallei, is an important cause of pneumonia, skin infection, sepsis, and death in Southeast Asia and Australia, but is exceedingly rare in North America. Pulmonary melioidosis typically presents as acute bacterial pneumonia or cavitary lung lesions resembling tuberculosis. We report melioidosis in a 70-year-old active smoker from Mexico with no history of travel to disease-endemic areas. The patient presented with a left supraclavicular abscess and a non-cavitary, left lung mass encasing a pulmonary vein. Incision and drainage of the patient's subcutaneous abscess isolated B. pseudomallei, and fine-needle aspiration of enlarged mediastinal lymph nodes revealed the presence of intracellular gram-negative bacilli with no evidence of malignancy. Biochemical tests determined that the strain the patient acquired from Mexico is identical to only 1 other isolate from Thailand. This report highlights the blurring epidemiological borders of this organism, its rare presentation mimicking lung malignancy, and an aggressive antimicrobial treatment that resulted in resolution of the patient's symptoms.

A case of bacterial peritonitis caused by Roseomonas mucosa in a patient undergoing continuous ambulatory peritoneal dialysis.

PubMed

Matsukuma, Yuta; Sugawara, Koji; Shimano, Shota; Yamada, Shunsuke; Tsuruya, Kazuhiko; Kitazono, Takanari; Higashi, Harumichi

2014-11-01

Bacterial peritonitis remains a life-threatening complication of peritoneal dialysis (PD). Roseomonas is a bacterial genus of pink-pigmented, oxidized, gram-negative coccobacilli that was first named in 1993. Importantly, Roseomonas mucosa exhibits antibiotic resistance, with significant resistance to cephalosporin, which is often selected as an empirical antibiotic regimen for peritonitis in PD patients. We herein report the case of a PD patient with bacterial peritonitis caused by Roseomonas mucosa that was fortunately identified using 16S rRNA gene sequencing and successfully treated with ciprofloxacin. Given that Roseomonas demonstrates resistance to a variety of antibiotics. The administration of empiric antibiotic therapy based on the recommendation of the International Society of Peritoneal Dialysis guidelines occasionally fails, leading to the aggravation of bacterial peritonitis. Hence, nephrologists should consider Roseomonas as one of the potential causative organisms of peritonitis, especially when gram-negative bacilli are resistant to cephalosporin and cannot be identified using standard laboratory methods.

Microbiological Etiologies of Pneumonia Complicating Stroke: A Systematic Review.

PubMed

Kishore, Amit K; Vail, Andy; Jeans, Adam R; Chamorro, Angel; Di Napoli, Mario; Kalra, Lalit; Langhorne, Peter; Roffe, Christine; Westendorp, Willeke; Nederkoorn, Paul J; Garau, Javier; van de Beek, Diederik; Montaner, Joan; Woodhead, Mark; Meisel, Andreas; Smith, Craig J

2018-06-18

Identifying the causal pathogens of pneumonia complicating stroke is challenging, and antibiotics used are often broad spectrum, without recourse to the microbiological cause. We aimed to review existing literature to identify organisms responsible for pneumonia complicating stroke, before developing a consensus-based approach to antibiotic treatment. A systematic literature review of multiple electronic databases using predefined search criteria was undertaken, in accordance with Cochrane and PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidance. Published studies of hospitalized adults with ischemic stroke, intracerebral hemorrhage, or both, which identified microbiological etiologies for pneumonia complicating stroke up to January 1, 2017, were considered. Analysis included summary statistics and random-effects meta-analysis where appropriate. Fifteen studies (40% ischemic stroke, 60% ischemic stroke and intracerebral hemorrhage) involving 7968 patients were included. Reported occurrence of pneumonia varied considerably between studies (2%-63%) with a pooled frequency of 23% (95% confidence interval, 14%-34%; I 2 =99%). Where reported (60%), the majority of pneumonia occurred within 1 week of stroke (78%). Reported frequency of positive culture data (15%-88%) varied widely. When isolated, aerobic Gram-negative bacilli (38%) and Gram-positive cocci (16%) were most frequently cultured; commonly isolated organisms included Enterobacteriaceae (21.8%: Klebsiella pneumoniae , 12.8% and Escherichia coli, 9%), Staphylococcus aureus (10.1%), Pseudomonas aeruginosa (6%), Acinetobacter baumanii (4.6%), and Streptococcus pneumoniae (3.5%). Sputum was most commonly used to identify pathogens, in isolation (40%) or in conjunction with tracheal aspirate (15%) or blood culture (20%). Although the analysis was limited by small and heterogeneous study populations, limiting determination of microbiological causality, this review suggests aerobic Gram-negative bacilli and Gram-positive cocci are frequently associated with pneumonia complicating stroke. This supports the need for appropriately designed studies to determine microbial cause and a consensus-based approach in antibiotic usage and further targeted antibiotic treatment trials for enhanced antibiotic stewardship. © 2018 American Heart Association, Inc.

[Infection control team (ICT) in cooperation with microbiology laboratories].

PubMed

Okazaki, Mitsuhiro

2012-10-01

Infection control as a medical safety measure is an important issue in all medical facilities. In order to tackle this measure, cooperation between the infection control team (ICT) and microbiological laboratory is indispensable. Multiple drug-resistant bacteria have shifted from Gram-positive bacteria to Gram-negative bacilli within the last ten years. There are also a variety of bacilli, complicating the examination method and test results further. Therefore, cooperation between the ICT and microbiological laboratory has become important to understand examination results and to use them. In order to maintain functional cooperation, explanatory and communicative ability between the microbiological laboratory and ICT is required every day. Such positive information exchange will develop into efficient and functional ICT activity.

[Utility of MALDI-TOF MS for the identification of anaerobic bacteria].

PubMed

Zárate, Mariela S; Romano, Vanesa; Nievas, Jimena; Smayevsky, Jorgelina

2014-01-01

The analysis by MALDI-TOF MS (Matrix-assited laser desorption/ionization time-of-flight mass spectrometry) has become a reference method for the identification of microorganisms in Clinical Microbiology. However, data on some groups of microorganisms are still controversial. The aim of this study is to determine the utility of MALDI-TOF MS for the identification of clinical isolates of anaerobic bacteria. One-hundred and six anaerobic bacteria isolates were analyzed by MALDI-TOF MS and by conventional biochemical tests. In those cases where identification by conventional methodology was not applicable or in the face of discordance between sequencing methodologies, 16 S rRNA gene sequence analysis was performed. The conventional method and MALDI-TOF MS agreed at genus and species level by 95.3 %. Concordance in gram-negative bacilli was 91.4% and 100% among gram-positive bacilli; there was also concordance both in the 8 isolates studied in gram-positive cocci and in the single gram-negative cocci included. The data obtained in this study demonstrate that MALDI-TOF MS offers the possibility of adequate identification of anaerobic bacteria. Copyright © 2014 Asociación Colombiana de Psiquiatría. Publicado por Elsevier España. All rights reserved.

Ants as vectors of pathogenic microorganisms in a hospital in São Paulo county, Brazil.

PubMed

Máximo, Heros J; Felizatti, Henrique L; Ceccato, Marcela; Cintra-Socolowski, Priscila; Beretta, Ana Laura R Zeni

2014-08-20

The present study aimed to identify and characterize the presence of bacteria carried by ants, and check the distribution of these ants in the physical confines of a medium-sized hospital in São Paulo county, Brazil. The ants were collected from March 2012 to February 2013. Attractive non-toxic baits were used to catch the ants, and the sectors considered for the study were medical wards, outdoor areas, obstetric unit, reception area, kitchen, surgical centres, paediatric clinic and intensive care unit. Captured ants were classified using taxonomic keys and subsequently immersed in Brain Heart Infusion broth. Paratrechina spp. and Monomorium floricola ants were found most frequently in the hospital. Ants had a high capacity for carrying bacteria, and the isolates comprised 68.8% Gram-positive, spore-producing bacilli (Bacillus spp. and Listeria spp.); 14.7% Gram-negative bacilli (Pseudomonas aeruginosa and Klebsiella spp.); and 16.4% Gram-positive cocci (Streptococcus spp. and Staphylococcus aureus). Among the areas being evaluated, the medical wards had the largest number of ants captured, and therefore the most bacteria. Ants in hospitals may carry both Gram-positive and Gram-negative bacteria, and methods of controlling urban ants should be adopted and strictly adhered to, to minimize the risk of infection in hospital patients.

Increasing resistant coagulase negative staphylococci in bovine clinical mastitis.

PubMed

Moniri, R; Dastehgoli, K; Akramian, A

2007-08-01

The aim of this study was to determine Coagulase Negative Staphylococci (CNS) and other bacteria for their resistance to antimicrobial agents approved for the control of pathogens involved in clinical bovine mastitis. This descriptive study was done on 106 milk samples obtained from clinical mastitis in dairy cattle husbandry from April 2006 through August 2006 in Kashan, Iran. From the total of 106 milk samples collected from clinical mastitis, 96 (90.6%) lead to positive culture. Coagulase negative Staphylococci isolated in 51 out of 96 samples (53.1%), Staphylococcus aureus isolated in 21 out of 96 (21.9%), gram negative bacilli isolated in 14 out of 96 (14.6%) and Enterococci isolated in 4 (4.2%). The highest rate of resistant CNS observed to penicillin (56.6%) and the highest rate of sensitivity to enrofloxacin 100%, followed by kanamycin, streptomycin and neomycin, 92.2, 82.3 and 82.3%, respectively. The highest rate of resistance S. aureus exhibited to penicillin (66.6%); while the highest rate of sensitivity showed to trimethoprim-sulphamethoxasole (81%), followed by kanamycin and enrofloxacin both at 76.2%. The highest rate of resistance gram negative bacilli exhibited to ampicillin and erythromycin at 71.4%. Their highest rate of sensitivity observed to enrofloxacin (78.6%), followed by kanamycin, (71.4%). In recent years, CNS is emerging as important minor mastitis pathogens and can be the cause of substantial economic losses. The high resistance rate to penicillin and other antibiotics found in this study emphasize the importance of identification of CNS when a bovine clinical mastitis is present.

Emergence of colistin resistance in the largest university hospital complex of São Paulo, Brazil, over five years.

PubMed

Rossi, Flávia; Girardello, Raquel; Cury, Ana Paula; Di Gioia, Thais Sabato Romano; Almeida, João Nóbrega de; Duarte, Alberto José da Silva

Colistin resistance involving Gram-negative bacilli infections is a challenge for health institutions around of the world. Carbapenem-resistance among these isolates makes colistin the last therapeutic option for this treatment. Colistin resistance among Enterobacteriaceae, Acinetobacter spp., and Pseudomonas spp. was evaluated between 2010 and 2014 years, at Hospital das Clínicas, São Paulo, Brazil. Over five years 1346 (4.0%) colistin resistant Gram-negative bacilli were evaluated. Enterobacteriaceae was the most frequent (86.1%) pathogen isolated, followed by Acinetobacter spp. (7.6%), and Pseudomonas spp. (6.3%). By temporal analysis there was a trend for an increase of colistin resistance among Enterobacteriaceae, but not among non-fermentative isolates. Among 1346 colistin resistant isolates, carbapenem susceptibility was observed in 21.5%. Colistin resistance in our hospital has been alarmingly increased among Klebsiella pneumoniae isolates in both KPC positive and negative, thus becoming a therapeutic problem. Copyright © 2016 Sociedade Brasileira de Infectologia. Published by Elsevier Editora Ltda. All rights reserved.

A single-tube screen for Salmonella and Shigella.

PubMed

Procop, Gary W; Wallace, Jacqueline D; Tuohy, Marion J; Lasalvia, Margret M; Addison, Rachel M; Reller, L Barth

2008-08-01

Salmonella and Shigella species are routinely sought in stool specimens submitted for culture. It is a common practice to screen lactose-negative colonies by using triple sugar iron agar, lysine iron agar, and Christensen urea agar to determine if further identification is necessary. We designed and evaluated a novel combination of media, which are layered in a single tube, for screening isolates suspected to possibly represent Salmonella or Shigella. We tested this media combination with 106 Salmonella, 56 Shigella, and 56 other gram-negative bacilli. All Salmonella and Shigella isolates tested were appropriately characterized as possible Salmonella or Shigella by using an algorithm developed for use with this media combination. Similarly, 53 (95%) of 56 other gram-negative bacilli were appropriately screened as non -Salmonella and non -Shigella isolates. This unique media combination provides the most important biochemical reactions needed to screen for Salmonella and Shigella in a single-tube format, which decreases labor by two thirds (ie, 1 tube is inoculated vs 3).

Diagnosis and treatment of bacterial prostatitis.

PubMed

Videčnik Zorman, Jerneja; Matičič, Mojca; Jeverica, Samo; Smrkolj, Tomaž

2015-01-01

Prostate inflammation is a common syndrome, especially in men under 50. It usually presents with voiding symptoms and pain in the genitourinary area, and sometimes as sexual dysfunction. Based on clinical and laboratory characteristics, prostatitis is classified as acute bacterial prostatitis, chronic bacterial prostatitis, chronic inflammatory and non-inflammatory prostatitis or chronic pelvic pain syndrome, and asymptomatic inflammatory prostatitis. Bacterial prostatitis is most often caused by infection with uropathogens, mainly Gram-negative bacilli, but Gram-positive and atypical microorganisms have also been identified as causative organisms of chronic prostatitis. According to reports by several authors, Chlamydia trachomatis and Trichomonas vaginalis are some of the most common pathogens, making chronic prostatitis a sexually transmitted disease. Diagnosis and treatment of acute and chronic bacterial prostatitis in particular can be challenging.

Nosocomial bloodstream infections in a Turkish university hospital: study of Gram-negative bacilli and their sensitivity patterns.

PubMed

Köseoğlu , O; Kocagöz, S; Gür, D; Akova, M

2001-06-01

Treatment of nosocomial bacteraemia is usually governed by the surveillance results of the particular unit. Such results are especially important when antimicrobial resistance rates are high. Multiresistant isolates including Gram-negatives producing extended-spectrum beta-lactamases have been frequently reported in tertiary care units in Turkey. In this study, antimicrobial susceptibilities of Gram-negative blood isolates (n=348) were determined by microbroth dilution tests. The results showed carbapenems (meropenem and imipenem) to be uniformly more potent in vitro than any other drug against the Enterobacteriaceae. Quinolone antibiotics were more active in vitro than aminoglycosides against a range of bacteria. Gram-negative bloodstream isolates were highly resistant to many antimicrobial agents in the hospital. In order to prevent hospital infection and antimicrobial resistance, surveillance of aetiological agents must be performed regularly.

Port type is a possible risk factor for implantable venous access port-related bloodstream infections and no sign of local infection predicts the growth of gram-negative bacilli.

PubMed

Hsu, Jui-Feng; Chang, Hsu-Liang; Tsai, Ming-Ju; Tsai, Ying-Ming; Lee, Yen-Lung; Chen, Pei-Huan; Fan, Wen-Chieh; Su, Yu-Chung; Yang, Chih-Jen

2015-09-30

Implantable venous access port (IVAP)-related blood stream infections (BSIs) are one of the most common complications of implantable venous ports. The risk factors and pathogens for IVAP-related BSIs are still controversial. We retrospectively reviewed all patients who received IVAPs at a Hospital in Taiwan from January 1, 2011 to June 31, 2014. Two types of venous port, BardPort® 6.6 fr (Bard port) and Autosuture Chemosite® 7.5 fr (TYCO port) were used. All patients with clinically proven venous port-related BSIs were enrolled. A total of 552 patients were enrolled. There were 34 episodes of IVAP-related BSIs during the study period for a total incidence of 0.177 events/1000 catheter days. Port type (TYCO vs. Bard, HR = 7.105 (95% confidence interval (CI), 1.688-29.904), p = 0.0075), age > 65 years (HR = 2.320 (95 % CI, 1.179-4.564), p = 0.0148), and lung cancer (HR = 5.807 (95% CI, 2.946-11.447), p < 0.001) were risk factors for port infections. We also found that no local sign of infection was significantly associated with the growth of gram-negative bacilli (p = 0.031). TYCO venous ports, age > 65 years, and lung cancer were all significant risk factors for IVAP-related BSIs, and no sign of infection was significantly associated with the growth of gram-negative bacilli.

Patient cytokine response in transfusion-associated sepsis.

PubMed Central

McAllister, S K; Bland, L A; Arduino, M J; Aguero, S M; Wenger, P N; Jarvis, W R

1994-01-01

Cytokine concentrations in plasma from patients transfused with packed erythrocytes contaminated with gram-negative bacilli were measured. Cytokine concentrations in posttransfusion plasma were significantly elevated. A difference in cytokine patterns between survivors and a nonsurvivor was observed. PMID:8168982

Multifocal suppurative granuloma caused by Actinobacillus lignieresii in the peritoneum of a beef steer

PubMed Central

KASUYA, Kazufumi; MANCHANAYAKE, Tilusha; UENOYAMA, Kei; KAWA, Sayaka; TAKAYAMA, Kou; IMAI, Naoto; SHIBAHARA, Tomoyuki

2016-01-01

An imported crossbred Angus beef steer aged eight to twelve months died suddenly on the eighth day of a quarantine period in Japan. Gross examination showed the peritoneum and mesentery consisted of numerous nodules of various sizes. Histological examination revealed chronic suppurative granulomatous peritonitis with eosinophilic rosettes surrounding colonies of Gram-negative bacilli. The bacteria isolated from the nodules were confirmed to be Actinobacillus lignieresii based on the results of 16S rRNA gene sequencing and immunohistochemistry. Antibiotic sensitivity testing showed that the isolate was resistant to penicillin. Thus, a diagnosis of atypical actinobacillosis caused by A. lignieresii was made. PMID:27773882

Multifocal suppurative granuloma caused by Actinobacillus lignieresii in the peritoneum of a beef steer.

PubMed

Kasuya, Kazufumi; Manchanayake, Tilusha; Uenoyama, Kei; Kawa, Sayaka; Takayama, Kou; Imai, Naoto; Shibahara, Tomoyuki

2017-01-20

An imported crossbred Angus beef steer aged eight to twelve months died suddenly on the eighth day of a quarantine period in Japan. Gross examination showed the peritoneum and mesentery consisted of numerous nodules of various sizes. Histological examination revealed chronic suppurative granulomatous peritonitis with eosinophilic rosettes surrounding colonies of Gram-negative bacilli. The bacteria isolated from the nodules were confirmed to be Actinobacillus lignieresii based on the results of 16S rRNA gene sequencing and immunohistochemistry. Antibiotic sensitivity testing showed that the isolate was resistant to penicillin. Thus, a diagnosis of atypical actinobacillosis caused by A. lignieresii was made.

Microbiology and Epidemiology of Legionnaire's Disease.

PubMed

Burillo, Almudena; Pedro-Botet, María Luisa; Bouza, Emilio

2017-03-01

Legionnaire's disease (LD) is the pneumonic form of legionellosis caused by aerobic gram-negative bacilli of the genus Legionella. Individuals become infected when they inhale aerosolized water droplets contaminated with Legionella species. Forty years after the identification of Legionella pneumophila as the cause of the 1976 pneumonia outbreak in a hotel in Philadelphia, we have non-culture-based diagnostic tests, effective antibiotics, and preventive measures to handle LD. With a mortality rate still around 10%, underreporting, and sporadic outbreaks, there is still much work to be done. In this article, the authors review the microbiology, laboratory diagnosis, and epidemiology of LD. Copyright © 2016 Elsevier Inc. All rights reserved.

Comparison of methods for the identification of microorganisms isolated from blood cultures.

PubMed

Monteiro, Aydir Cecília Marinho; Fortaleza, Carlos Magno Castelo Branco; Ferreira, Adriano Martison; Cavalcante, Ricardo de Souza; Mondelli, Alessandro Lia; Bagagli, Eduardo; da Cunha, Maria de Lourdes Ribeiro de Souza

2016-08-05

Bloodstream infections are responsible for thousands of deaths each year. The rapid identification of the microorganisms causing these infections permits correct therapeutic management that will improve the prognosis of the patient. In an attempt to reduce the time spent on this step, microorganism identification devices have been developed, including the VITEK(®) 2 system, which is currently used in routine clinical microbiology laboratories. This study evaluated the accuracy of the VITEK(®) 2 system in the identification of 400 microorganisms isolated from blood cultures and compared the results to those obtained with conventional phenotypic and genotypic methods. In parallel to the phenotypic identification methods, the DNA of these microorganisms was extracted directly from the blood culture bottles for genotypic identification by the polymerase chain reaction (PCR) and DNA sequencing. The automated VITEK(®) 2 system correctly identified 94.7 % (379/400) of the isolates. The YST and GN cards resulted in 100 % correct identifications of yeasts (15/15) and Gram-negative bacilli (165/165), respectively. The GP card correctly identified 92.6 % (199/215) of Gram-positive cocci, while the ANC card was unable to correctly identify any Gram-positive bacilli (0/5). The performance of the VITEK(®) 2 system was considered acceptable and statistical analysis showed that the system is a suitable option for routine clinical microbiology laboratories to identify different microorganisms.

Microbiology of liver abscesses and the predictive value of abscess gram stain and associated blood cultures.

PubMed

Chemaly, Roy F; Hall, Gerri S; Keys, Thomas F; Procop, Gary W

2003-08-01

Although rare, pyogenic liver abscesses are potentially fatal. We evaluated the predictive value of Gram stain of liver abscess aspirates and temporally associated blood cultures. Gram stains detected bacteria in 79% of the liver abscesses tested. The sensitivity and specificity of Gram stain of the liver abscesses were 90% and 100% for Gram-positive cocci (GPC) and 52% and 94% for Gram-negative bacilli (GNB). The sensitivities of the blood cultures for any GPC and GNB present in the liver abscess were 30% and 39%, respectively. Although, Gram stains and blood cultures offer incomplete detection of the microbial contents of pyogenic liver abscesses, both tests should always accompany liver abscess cultures.

9 CFR 145.1 - Definitions.

Code of Federal Regulations, 2011 CFR

2011-01-01

.... Domesticated. Propagated and maintained under the control of a person. Equivalent or equivalent requirements... provisions of the Plan; (2) As applied to non-Plan but equivalent State poultry improvement programs. The... of this chapter, those organisms which are gram negative, non spore-forming bacilli, which ferment...

9 CFR 145.1 - Definitions.

Code of Federal Regulations, 2010 CFR

2010-01-01

.... Domesticated. Propagated and maintained under the control of a person. Equivalent or equivalent requirements... provisions of the Plan; (2) As applied to non-Plan but equivalent State poultry improvement programs. The... of this chapter, those organisms which are gram negative, non spore-forming bacilli, which ferment...

Antimicrobial susceptibility of Gram-negative bacteria causing intra-abdominal infections in China: SMART China 2011.

PubMed

Zhang, Hui; Yang, Qiwen; Xiao, Meng; Chen, Minjun; Badal, Robert E; Xu, Yingchun

2014-01-01

The Study for Monitoring Antimicrobial Resistance Trends program monitors the activity of antibiotics against aerobic and facultative Gram-negative bacilli (GNBs) from intra-abdominal infections (IAIs) in patients worldwide. In 2011, 1 929 aerobic and facultative GNBs from 21 hospitals in 16 cities in China were collected. All isolates were tested using a panel of 12 antimicrobial agents, and susceptibility was determined following the Clinical Laboratory Standards Institute guidelines. Among the Gram-negative pathogens causing IAIs, Escherichia coli (47.3%) was the most commonly isolated, followed by Klebsiella pneumoniae (17.2%), Pseudomonas aeruginosa (10.1%), and Acinetobacter baumannii (8.3%). Enterobacteriaceae comprised 78.8% (1521/1929) of the total isolates. Among the antimicrobial agents tested, ertapenem and imipenem were the most active agents against Enterobacteriaceae, with susceptibility rates of 95.1% and 94.4%, followed by amikacin (93.9%) and piperacillin/tazobactam (87.7%). Susceptibility rates of ceftriaxone, cefotaxime, ceftazidime, and cefepime against Enterobacteriaceae were 38.3%, 38.3%, 61.1%, and 50.8%, respectively. The leastactive agent against Enterobacteriaceae was ampicillin/sulbactam (25.9%). The extended-spectrum β-lactamase (ESBL) rates among E. coli, K. pneumoniae, Klebsiella oxytoca, and Proteus mirabilis were 68.8%, 38.1%, 41.2%, and 57.7%, respectively. Enterobacteriaceae were the major pathogens causing IAIs, and the most active agents against the study isolates (including those producing ESBLs) were ertapenem, imipenem, and amikacin. Including the carbapenems, most agents exhibited reduced susceptibility against ESBL-positive and multidrug-resistant isolates.

Athlete's foot caused by pseudomonas aeruginosa.

PubMed

Abramson, C

1983-01-01

An enzymatically active pigment-producing clinical isolate of Pseudomonas aeruginosa was found to produce a diffusible antifungal product that was shown to be inhibitory to the growth of several dermatophytes, specifically, Trichophyton rubrum, Trichophyton mentagrophytes, Microsporum gypseum, and Microsporum audouini. In this study, Trichophyton rubrum was used as the test organism. The antifungal product was partially purified by Sephadex column chromatography and was found to be stable at 5 degrees, 25 degrees, and 37 degrees C. Several investigators have alluded to the fact that as asymptomatic cases of dermatophytosis simplex progress to symptomatic dermatophytosis complex, the bacterial profile changes from a gram-positive bacterial ecosystem to a gram-negative bacterial over-growth. The primary event in the pathogenesis of interdigital athlete's foot is the invasion of the horny layer by dermatophytes. This presents as a mild to moderate scaly lesion and is asymptomatic. As a result of predisposing factors, such as hyperhidrosis, occlusion by tight shoes, minute abrasions due to friction, and fungal-infected skin surfaces, dynamic overgrowth of opportunistic gram-negative bacilli prevails. As the gram-negative population increases, the recovery of dermatophytes dramatically diminishes, until a point is reached when no dermatophytes can be recovered from clinically symptomatic tinea pedis. Pseudomonas aeruginosa is inhibiting its fungal competitor Trichophyton rubrum by producing a diffusible antifungal agent into the infectious environment of the intertriginous foot lesion. Clinically, the patient is diagnosed as having tinea pedis; laboratory culture for fungus and KOH are negative, and what was a paradox just a few years ago can currently be identified and treated appropriately as gram-negative athlete's foot.

[Antimicrobial susceptibility testing in clinically relevant non-fermenting gram-negative bacilli: recommendations from the Antimicrobial Agents Subcommittee of the Sociedad Argentina de Bacteriología, Micología y Parasitología Clínicas, Asociación Argentina de Microbiología].

PubMed

Radice, Marcela; Marín, Marcelo; Giovanakis, Marta; Vay, Carlos; Almuzara, Marisa; Limansky, Adriana; Casellas, José M; Famiglietti, Angela; Quinteros, Mirta; Bantar, Carlos; Galas, Marcelo; Kovensky Pupko, Jaime; Nicola, Federico; Pasterán, Fernando; Soloaga, Rolando; Gutkind, Gabriel

2011-01-01

This document contains the recommendations for antimicrobial susceptibility testing of the clinically relevant non-fermenting gram-negative bacilli (NFGNB), adopted after conforming those from international committees to the experience of the Antimicrobial Agents Subcommittee members and invited experts. This document includes an update on NFGNB classification and description, as well as some specific descriptions regarding natural or frequent antimicrobial resistance and a brief account of associated resistance mechanisms. These recommendations not only suggest the antimicrobial drugs to be evaluated in each case, but also provide an optimization of the disk diffusion layout and a selection of results to be reported. Finally, this document also includes a summary of the different methodological approaches that may be used for detection and confirmation of emerging b-lactamases, such as class A and B carbapenemases.

Evaluation of Five Chromogenic Agar Media and the Rosco Rapid Carb Screen Kit for Detection and Confirmation of Carbapenemase Production in Gram-Negative Bacilli

PubMed Central

Gilmour, Matthew W.; DeGagne, Pat; Nichol, Kim; Karlowsky, James A.

2014-01-01

An efficient workflow to screen for and confirm the presence of carbapenemase-producing Gram-negative bacilli was developed by evaluating five chromogenic screening agar media and two confirmatory assays, the Rapid Carb screen test (Rosco Diagnostica A/S, Taastrup, Denmark) and the modified Hodge test. A panel of 150 isolates was used, including 49 carbapenemase-producing isolates representing a variety of β-lactamase enzyme classes. An evaluation of analytical performance, assay cost, and turnaround time indicated that the preferred workflow (screening test followed by confirmatory testing) was the chromID Carba agar medium (bioMérieux, Marcy l'Étoile, France), followed by the Rapid Carb screen test, yielding a combined sensitivity of 89.8% and a specificity of 100%. As an optional component of the workflow, a determination of carbapenemase gene class via molecular means could be performed subsequent to confirmatory testing. PMID:25355764

Infections in cancer patients on a protected environment-prophylactic antibiotic program.

PubMed

Bodey, G P; Rodriguez, V

1975-10-01

A total of 102 studies were conducted on 89 patients receiving cancer chemotherapy while on a protected environment-prophylactic antibiotic program. Major infections occurred during 22 studies. The majority of both minor and major infections originated during the first five weeks after the patients entered the protected environment units. The frequency of infectious complications was inversely related to the circulating neutrophil count. The majority of infections were cases of cellulitis, pharyngitis, pneumonia and septicemia. Most of the infections were caused by gram-negative bacilli. The majority of organisms causing infection had persisted in the patients after their entry into the protected environment units despite the use of prophylactic antibiotics.

The abundance and diversity of heterotrophic bacteria as a function of harvesting frequency of duckweed (Lemna minor L.) in recirculating aquaculture systems.

PubMed

Ardiansyah, A; Fotedar, R

2016-07-01

Duckweed (Lemna minor L.) is a potential biofilter for nutrient removal and acts as a substrate for heterotrophic bacteria in recirculating aquaculture systems (RAS). Here, we determined the effects of harvesting frequency of duckweed on heterotrophic bacteria in RAS. Twelve independent RAS consisting of fish-rearing tank, biofilter tank and waste-collection tank were used to study the interactions between duckweed harvest frequencies up to 6 days and the composition, abundance and diversity of heterotrophic bacteria. After 36 days, heterotrophic bacteria in the biofilter tank were primarily Gram-negative cocci or ovoid, coccobacilli, Gram-negative bacilli and Gram-positive bacilli. Most bacterial genera were Bacillus and Pseudomonas while the least common was Acinetobacter. Duckweed harvested after every 2 days produced the highest specific growth rates (SGR) and total harvested biomass of duckweed, but harboured less abundant bacteria, whereas 6-day harvests had a higher growth index (GI) of duckweed than 2-day harvests, but caused a poor relationship between SGR and biomass harvest with the abundance and diversity of heterotrophic bacteria. Stronger correlations (R(2)  > 0·65) between duckweed SGR and biomass harvest with the heterotrophic bacteria diversity were observed at 4-day harvest frequency and the control. This study provides significant information on the interaction between the harvest frequency of duckweed and the composition, abundance and diversity of heterotrophic bacteria in recirculating aquaculture systems (RAS). Different harvest frequencies significantly influence the abundance and diversity of heterotrophic bacteria, which in turn may influence the nitrogen uptake efficiency of the system. The research is useful in improving the efficiency of removing nitrogenous metabolites in RAS directly by the duckweed and associated heterotrophic bacteria. © 2016 The Society for Applied Microbiology.

[News of antibiotic resistance among Gram-negative bacilli in Algeria].

PubMed

Baba Ahmed-Kazi Tani, Z; Arlet, G

2014-06-01

Antibiotic resistance has become a major public health problem in Algeria. Indeed the past decade, we have seen a significant increase in resistance to antibiotics especially in Gram-negative bacilli. Resistance to β-lactams in enterobacteria is dominated by the production of ESBL CTX-M-3 and CTX-M-15. The strains producing these enzymes are often the cause of potentially serious infections in both hospital and community settings. Identified plasmid cephalosporinases are CMY-2, CMY-12 and DHA-1. The isolation of strains of Enterobacteriaceae and Pseudomonas aeruginosa producing carbapenemases is rare in Algeria. Some Enterobacteriaceae producing OXA-48 or VIM-19 have been reported; so far, only VIM-2 has been identified in P. aeruginosa. However, the situation regarding the strains of Acinetobacter baumannii resistant to carbapenemases seems to be more disturbing. The carbapenemase OXA-23 is the most common and seems to be endemic in the north. The carbapenemase NDM-1 has also been identified. Resistance to aminoglycosides is marked by the identification armA gene associated with blaCTX-M genes in strains of Salmonella sp. Several other resistance genes have been identified sporadically in strains of Enterobacteriaceae, P. aeruginosa and A. baumannii. Resistance genes to fluoroquinolones are more recent identification in Algeria. The most common are the Qnr determinants followed by the bifunctional enzyme AAC[6']-Ib-cr. Resistance to sulfonamides and trimethoprim was also reported in Enterobacteriaceae strains in the west of the country. Copyright © 2014. Published by Elsevier SAS.

Epidemiology and trends in the antibiotic susceptibilities of Gram-negative bacilli isolated from patients with intra-abdominal infections in the Asia-Pacific region, 2010-2013.

PubMed

Chang, Ya-Ting; Coombs, Geoffrey; Ling, Thomas; Balaji, V; Rodrigues, Camilla; Mikamo, Hiroshige; Kim, Min-Ja; Rajasekaram, Datin Ganeswrie; Mendoza, Myrna; Tan, Thean Yen; Kiratisin, Pattarachai; Ni, Yuxing; Barry, Weinman; Xu, Yingchun; Chen, Yen-Hsu; Hsueh, Po-Ren

2017-06-01

This study was conducted to investigate the epidemiology and antimicrobial susceptibility patterns of Gram-negative bacilli (GNB) isolated from intra-abdominal infections (IAIs) in the Asia-Pacific region (APR) from 2010-2013. A total of 17 350 isolates were collected from 54 centres in 13 countries in the APR. The three most commonly isolated GNB were Escherichia coli (46.1%), Klebsiella pneumoniae (19.3%) and Pseudomonas aeruginosa (9.8%). Overall, the rates of extended-spectrum β-lactamase (ESBL)-producing E. coli and K. pneumoniae were 38.2% and 24.3%, respectively, and they were highest in China (66.6% and 38.7%, respectively), Thailand (49.8% and 36.5%, respectively) and Vietnam (47.9% and 30.4%, respectively). During 2010-2013, the rates of ESBL-producing E. coli and K. pneumoniae isolates causing community-associated (CA) IAIs (collected <48 h after admission) were 26.0% and 13.5%, respectively, and those causing hospital-associated (HA) IAIs were 48.0% and 30.6%, respectively. Amikacin, ertapenem and imipenem were the most effective agents against ESBL-producing isolates. Piperacillin/tazobactam displayed good in vitro activity (91.4%) against CA ESBL-producing E. coli. For other commonly isolated Enterobacteriaceae, fluoroquinolones, cefepime and carbapenems exhibited better in vitro activities than third-generation cephalosporins. Amikacin possessed high in vitro activity against all GNB isolates (>80%) causing IAIs, except for Acinetobacter calcoaceticus-baumannii (ACB) complex (30.9% for HA-IAI isolates). All of the antimicrobial agents tested exhibited <45% in vitro activity against ACB complex. Antimicrobial resistance is a persistent threat in the APR and continuous monitoring of evolutionary trends in the susceptibility patterns of GNB causing IAIs in this region is mandatory. Copyright © 2017 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.

Activities of fosfomycin, tigecycline, colistin, and gentamicin against extended-spectrum-β-lactamase-producing Escherichia coli in a foreign-body infection model.

PubMed

Corvec, Stéphane; Furustrand Tafin, Ulrika; Betrisey, Bertrand; Borens, Olivier; Trampuz, Andrej

2013-03-01

Limited antimicrobial agents are available for the treatment of implant-associated infections caused by fluoroquinolone-resistant Gram-negative bacilli. We compared the activities of fosfomycin, tigecycline, colistin, and gentamicin (alone and in combination) against a CTX-M15-producing strain of Escherichia coli (Bj HDE-1) in vitro and in a foreign-body infection model. The MIC and the minimal bactericidal concentration in logarithmic phase (MBC(log)) and stationary phase (MBC(stat)) were 0.12, 0.12, and 8 μg/ml for fosfomycin, 0.25, 32, and 32 μg/ml for tigecycline, 0.25, 0.5, and 2 μg/ml for colistin, and 2, 8, and 16 μg/ml for gentamicin, respectively. In time-kill studies, colistin showed concentration-dependent activity, but regrowth occurred after 24 h. Fosfomycin demonstrated rapid bactericidal activity at the MIC, and no regrowth occurred. Synergistic activity between fosfomycin and colistin in vitro was observed, with no detectable bacterial counts after 6 h. In animal studies, fosfomycin reduced planktonic counts by 4 log(10) CFU/ml, whereas in combination with colistin, tigecycline, or gentamicin, it reduced counts by >6 log(10) CFU/ml. Fosfomycin was the only single agent which was able to eradicate E. coli biofilms (cure rate, 17% of implanted, infected cages). In combination, colistin plus tigecycline (50%) and fosfomycin plus gentamicin (42%) cured significantly more infected cages than colistin plus gentamicin (33%) or fosfomycin plus tigecycline (25%) (P < 0.05). The combination of fosfomycin plus colistin showed the highest cure rate (67%), which was significantly better than that of fosfomycin alone (P < 0.05). In conclusion, the combination of fosfomycin plus colistin is a promising treatment option for implant-associated infections caused by fluoroquinolone-resistant Gram-negative bacilli.

9 CFR 145.1 - Definitions.

Code of Federal Regulations, 2014 CFR

2014-01-01

... Agriculture. Domesticated. Propagated and maintained under the control of a person. Equivalent or equivalent... bacilli. For the purpose of this chapter, those organisms which are gram negative, non spore-forming... classification on one farm; (2) As applied to disease control. All of the poultry on one farm except that, at the...

9 CFR 145.1 - Definitions.

Code of Federal Regulations, 2012 CFR

2012-01-01

... Agriculture. Domesticated. Propagated and maintained under the control of a person. Equivalent or equivalent... bacilli. For the purpose of this chapter, those organisms which are gram negative, non spore-forming... classification on one farm; (2) As applied to disease control. All of the poultry on one farm except that, at the...

9 CFR 145.1 - Definitions.

Code of Federal Regulations, 2013 CFR

2013-01-01

... Agriculture. Domesticated. Propagated and maintained under the control of a person. Equivalent or equivalent... bacilli. For the purpose of this chapter, those organisms which are gram negative, non spore-forming... classification on one farm; (2) As applied to disease control. All of the poultry on one farm except that, at the...

Prosthetic joint infection caused by Trueperella bernardiae.

PubMed

Gilarranz, Raul; Chamizo, Francisco; Horcajada, Iballa; Bordes-Benítez, Ana

2016-09-01

Trueperella bernardiae is a Gram-positive coryneform bacilli which role as human pathogen is unknown because it has been usually considered a contaminant. Furthermore its identification by biochemical test was difficult. We describe a prosthetic joint infection in a women who years ago underwent a total knee replacement with superinfection and necrosis of the patellar tendon as major complications. In the sample of synovial fluid collected grew a gram-positive bacilli which was identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) as T. bernardiae. The patient was treated with ciprofloxacin and currently preserves the prosthesis without signs of infection. Copyright © 2016 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Novel fastidious, partially acid-fast, anaerobic Gram-positive bacillus associated with abscess formation and recovered from multiple medical centers.

PubMed

Harrington, S M; Bell, M; Bernard, K; Lagacé-Wiens, P; Schuetz, A N; Hartman, B; McQuiston, J R; Wilson, D; Lasalvia, M; Ng, B; Richter, S; Taege, A

2013-11-01

We report a novel anaerobe causing abscess in four patients at three hospitals. In the clinical specimen, bacilli were branching, Gram positive, and acid fast. The organism grew slowly and was not identified by 16S rRNA sequencing. Our findings support the description of a new genus and species of the suborder Corynebacterineae.

Post-traumatic infection of the lower limb caused by rare Enterobacteriaceae and Mucorales in a young healthy male.

PubMed

Corti, Giampaolo; Mondanelli, Nicola; Losco, Michele; Bartolini, Laura; Fontanelli, Alessandra; Paradisi, Franco

2009-03-01

Enterobacter amnigenus and Leclercia adecarboxylata are gram-negative aerobic bacilli of the family Enterobacteriaceae that have been isolated from water and, rarely, from various clinical specimens. Absidia is a filamentous fungus of the class Zygomycetes that is ubiquitous in nature and can cause infection, primarily in immunocompromised hosts. Here, we describe an infection of the left lower limb caused by E. amnigenus and L. adecarboxylata with subsequent isolation of Absidia spp. in a patient with multiple traumatic injuries after a major motor vehicle accident. The severity of the clinical picture made amputation necessary, despite aggressive anti-infective therapy with both antibacterial and antifungal agents. Prompt diagnosis and management are mandatory in order to minimize morbidity and even mortality, and reduce the social and economic cost.

Nosocomial Bloodstream Infections in Brazilian Pediatric Patients: Microbiology, Epidemiology, and Clinical Features

PubMed Central

Pereira, Carlos Alberto Pires; Marra, Alexandre R.; Camargo, Luis Fernando Aranha; Pignatari, Antônio Carlos Campos; Sukiennik, Teresa; Behar, Paulo Renato Petersen; Medeiros, Eduardo Alexandrino Servolo; Ribeiro, Julival; Girão, Evelyne; Correa, Luci; Guerra, Carla; Carneiro, Irna; Brites, Carlos; Reis, Marise; de Souza, Marta Antunes; Tranchesi, Regina; Barata, Cristina U.; Edmond, Michael B.

2013-01-01

Background Nosocomial bloodstream infections (nBSIs) are an important cause of morbidity and mortality and are the most frequent type of nosocomial infection in pediatric patients. Methods We identified the predominant pathogens and antimicrobial susceptibilities of nosocomial bloodstream isolates in pediatric patients (≤16 years of age) in the Brazilian Prospective Surveillance for nBSIs at 16 hospitals from 12 June 2007 to 31 March 2010 (Br SCOPE project). Results In our study a total of 2,563 cases of nBSI were reported by hospitals participating in the Br SCOPE project. Among these, 342 clinically significant episodes of BSI were identified in pediatric patients (≤16 years of age). Ninety-six percent of BSIs were monomicrobial. Gram-negative organisms caused 49.0% of these BSIs, Gram-positive organisms caused 42.6%, and fungi caused 8.4%. The most common pathogens were Coagulase-negative staphylococci (CoNS) (21.3%), Klebsiella spp. (15.7%), Staphylococcus aureus (10.6%), and Acinetobacter spp. (9.2%). The crude mortality was 21.6% (74 of 342). Forty-five percent of nBSIs occurred in a pediatric or neonatal intensive-care unit (ICU). The most frequent underlying conditions were malignancy, in 95 patients (27.8%). Among the potential factors predisposing patients to BSI, central venous catheters were the most frequent (66.4%). Methicillin resistance was detected in 37 S. aureus isolates (27.1%). Of the Klebsiella spp. isolates, 43.2% were resistant to ceftriaxone. Of the Acinetobacter spp. and Pseudomonas aeruginosa isolates, 42.9% and 21.4%, respectively, were resistant to imipenem. Conclusions In our multicenter study, we found a high mortality and a large proportion of gram-negative bacilli with elevated levels of resistance in pediatric patients. PMID:23861860

Moraxella osloensis: an unusual cause of central venous catheter infection in a cancer patient

PubMed Central

Hadano, Yoshiro; Ito, Kenta; Suzuki, Jun; Kawamura, Ichiro; Kurai, Hanako; Ohkusu, Kiyofumi

2012-01-01

Moraxella osloensis is a rare causative organism of infections in humans, with most cases reported in cancer patients. We report the case of a 67-year-old Japanese man with advanced cancer of the pancreatic head and multiple liver metastases who developed fever with chills. Blood culture was found to be positive for Gram-negative bacilli that were aerobic, oxidase-positive, and catalase-positive. M. osloensis was identified by 16 rRNA gene sequencing. Prompt control of the infection was achieved by treatment with cefepime for 14 days, without the need for removal of the central venous catheter. PMID:23109812

Moraxella osloensis: an unusual cause of central venous catheter infection in a cancer patient.

PubMed

Hadano, Yoshiro; Ito, Kenta; Suzuki, Jun; Kawamura, Ichiro; Kurai, Hanako; Ohkusu, Kiyofumi

2012-01-01

Moraxella osloensis is a rare causative organism of infections in humans, with most cases reported in cancer patients. We report the case of a 67-year-old Japanese man with advanced cancer of the pancreatic head and multiple liver metastases who developed fever with chills. Blood culture was found to be positive for Gram-negative bacilli that were aerobic, oxidase-positive, and catalase-positive. M. osloensis was identified by 16 rRNA gene sequencing. Prompt control of the infection was achieved by treatment with cefepime for 14 days, without the need for removal of the central venous catheter.

Cerebral hemorrhage in infective endocarditis caused by Actinobacillus actinomycetemcomitans.

PubMed

Lin, Gen-Min; Chu, Kai-Min; Juan, Chun-Jung; Chang, Feng-Yee

2007-11-01

Cerebral hemorrhage occurs rarely in endocarditis caused by Actinobacillus actinomycetemcomitans. A 51-year-old man with a prosthetic mitral valve, who had been prophylactically treated (7 years) with warfarin, presented with intermittent fever. On admission, a Levine grade II/VI systolic cardiac murmur was detected. A transthoracic echocardiogram was negative for valve vegetation. Cefepime (1 g every 8 hours) was administered intravenously. On day 4, culturing of Gram-negative bacilli from blood and a transesophageal echocardiogram revealed a small oscillating filament attached to lateral mitral prosthetic ring on the atrial side. Ceftriaxone (2 g once daily) was started. Gait instability and left-side weakness developed abruptly 2 weeks later; brain magnetic resonance imaging revealed a hematoma over the right parietal-occipital lobe. Ceftriaxone was adjusted to 2 g every 12 hours. Actinobacillus actinomycetemcomitans was identified 3 weeks later. Recovery was achieved, with significant interval improvement and resolution of the cerebral lesions evident on CT.

Complete Genome Sequences for 59 Burkholderia Isolates, Both Pathogenic and Near Neighbor

DOE PAGES

Johnson, Shannon L.; Bishop-Lilly, Kimberly A.; Ladner, Jason T.; ...

2015-04-30

The genus Burkholderia encompasses both pathogenic (including Burkholderia mallei and Burkholderia pseudomallei, U.S. Centers for Disease Control and Prevention Category B listed), and nonpathogenic Gram-negative bacilli. Presented in this document are full genome sequences for a panel of 59 Burkholderia strains, selected to aid in detection assay development.

Development of rapid phenotypic system for the identification of Gram-negative oxidase-positive bacilli in resource-limited settings.

PubMed

Kazmi, Mahmooda; Khan, Adnan; Kazmi, Shahana Urooj

2013-06-01

Rapid and accurate identification of bacterial pathogens is a fundamental goal of clinical microbiology. The diagnosis and surveillance of diseases is dependent, to a great extent, on laboratory services, which cannot function without effective reliable reagents and diagnostics. Despite the advancement in microbiology diagnosis globally, resourcelimited countries still struggle to provide an acceptable diagnosis quality which helps in clinical disease management and improve their mortality and morbidity data. During this study an indigenous product, Quick Test Strip (QTS) NE, was developed for the rapid identification of biochemically slower group of Gram-negative oxidase-positive bacilli that covers 19 different bacterial genera. Some of the members belonging to these groups are well-established human pathogens, e.g. various species of Vibrio, Pseudomonas, Burkholderia, Aeromonas, Achromobacter and Stenotrophomonas. This study also evaluates the performance of QTS-NE by comparing with genotypic characterization methods. A total of 232 clinical and reference bacterial isolates were tested by three different methods. QTSNE provides 100 percent concordant results with other rapid identification and molecular characterization methods and confirms the potential to be used in clinical diagnosis.

Identification of gram-negative bacilli using the Autobac IDX.

PubMed

Burdash, N M; Welborn, A L; Teti, G; Bannister, E R; Manos, J P

1985-01-01

The Autobac IDX is a new system for the rapid identification of clinically significant members of the Enterobacteriaceae and Aeromonas, Acinetobacter, Alcaligenes, Flavobacterium, Moraxella, and Pseudomonas species. The use of 18 differentially inhibitory compounds such as dyes and antibiotics along with a computerized algorithm based on a multivariate analysis provides the basis for the identification of 30 different groups of gram-negative bacilli. Required preliminary tests include observations on the presence or absence of swarming on a sheep blood agar plate and noting the following: growth, lactose fermentation, and bile precipitation from a MacConkey plate. Spot indole and spot oxidase tests must be performed as well. Identification by the Autobac IDX System takes 3-6 hr after completion of the preliminary tests. From a total of 403 isolates tested, the Autobac system agreed with the MicroID AND N/F systems on 382 identifications (94.8%). Four isolates, two Acinetobacter anitratus, one Serratia marcescens and one Moraxella osloensis could not be identified by IDX. Additional testing was required on 35 (8.7%) of the isolates.

[Test evaluation and strategy proposal to detect and to characterize carbapenemase-producing gram negative bacilli].

PubMed

Muñoz, Constanza; Zumarán, Cecilia; González, Tamara; Wozniak, Aniela; Castillo, Claudia; García, Patricia

2017-08-01

The detection of carbapenemase-producing gram negative bacilli is complicated, because there are available multiple options of test. The confirmation of the enzyme by molecular characterization is not available in all laboratories in our country. To propose a fast, efficient and simple strategy to detect and confirm CPB. 39 CPB isolates and 8 non-producing were used to evaluate the phenotypic test Carba NP, CarbAcineto NP and Blue-Carba, validating the test Xpert® Carba-R, to be used directly with bacterial colonies with conventional PCR. The sensitivity of Carba NP, CarbAcineto NP and Blue-Carba was 79,5; 87,2 y 84,6%, respectively; and specificity was 79.5; 87.2 and 84.6%, respectively. The limit of detection of Xpert® Carba-R was different for each carbapenemasa: 40.8 ufc/reaction to KPC and NDM and 30.6 ufc/reaction to VIM. On isolates with decreased susceptibility to carbapenems we propose to use as screening the test CarbAcineto NP, follow by Xpert®Carba-R to characterize the carbapenemase and adopt specific infection control measures.

Case of Mycobacterium tuberculosis meningitis: Gram staining as a useful initial diagnostic clue for tuberculous meningitis.

PubMed

Kawakami, Sayoko; Kawamura, Yasuyosi; Nishiyama, Kyouhei; Hatanaka, Hiroki; Fujisaki, Ryuichi; Ono, Yasuo; Miyazawa, Yukihisa; Nishiya, Hajime

2012-12-01

A 32-year-old man was admitted to our hospital because of fever, headache, and loss of consciousness. Four days before admission, he had had difficulty speaking. On the day of admission, his colleague had found him to be unconscious and lying on his back. He was admitted to our hospital. The temperature at the eardrum was 35.2°C. Neurologic evaluation was negative. Computed tomography (CT) scan of the brain showed slight ventricular enlargement bilaterally. An X-ray film of the chest showed no abnormality. On the second hospital day, neck stiffness was noted. The cerebrospinal fluid (CSF) contained 870 white cells/μl, most of which were neutrophils; the glucose level in the CSF was 10 mg/dl, and the protein level was 140 mg/dl. Stained smears of the CSF, including Gram staining and India-ink preparations, disclosed no microorganisms. Capsular antigen tests for several bacteria were negative. Antimicrobial agents were started. However, by changing the microscope focus slightly while viewing Gram stains of the CSF, we could see brightened and Gram-positive bacilli that had been phagocytosed by neutrophils. This finding suggested the presence of Mycobacterium tuberculosis. Ziehl-Neelsen staining of the CSF and gastric juice revealed anti-acid bacilli. Polymerase chain reaction for M. tuberculosis in the gastric juice was positive. This case showed that Gram staining could be useful as an initial adjunct for the diagnosis of tuberculous meningitis, particularly when the CSF shows predominantly neutrocytic pleocytosis, but no other evidence of bacterial meningitis.

[Anaerobic bacteria isolated from patients with suspected anaerobic infections].

PubMed

Ercis, Serpil; Tunçkanat, Ferda; Hasçelik, Gülşen

2005-10-01

The study involved 394 clinical samples sent to the Clinical Microbiology Laboratory of Hacettepe University Adult Hospital between January 1997 and May 2004 for anaerobic cultivation. Since multiple cultures from the same clinical samples of the same patient were excluded, the study was carried on 367 samples. The anaerobic cultures were performed in anaerobic jar using AnaeroGen kits (Oxoid, Basingstoke, U.K.) or GENbox (bioMérieux, Lyon, France). The isolates were identified by both classical methods and "BBL Crystal System" (Becton Dickinson, U.S.A.). While no growth was detected in 120 (32.7%) of the clinical samples studied, in 144 samples (39.2%) only aerobes, in 28 (7.6%) only anaerobes and in 75 (20.5%) of the samples both aerobes and anaerobes were isolated. The number of the anaerobic isolates was 217 from 103 samples with anaerobic growth. Of these 103 samples 15 showed single bacterial growth whereas in 88 samples multiple bacterial isolates were detected. Anaerobic isolates consisted of 92 Gram negative bacilli (Bacteroides spp. 50, Prevotella spp. 14, Porphyromonas spp. 10, Fusobacterium spp. 7, Tisierella spp. 2, unidentified 9), 57 Gram positive bacilli (Clostridium spp.17, Propionibacterium spp. 16, Lactobacillus spp. 8, Actinomyces spp. 5, Eubacterium spp. 2, Bifidobacterium adolescentis 1, Mobiluncus mulieris 1, unidentified nonspore forming rods 7), 61 Gram positive cocci (anaerobic cocci 44, microaerophilic cocci 17), and 7 Gram negative cocci (Veillonella spp.). In conclusion, in the samples studied with prediagnosis of anaerobic infection, Bacteroides spp. (23%) were the most common bacteria followed by anaerobic Gram positive cocci (20.3%) and Clostridium spp (7.8%).

[Bacteriologic study of abscesses caused by bites of snakes of the genus Bothrops].

PubMed

de Andrade, J G; Pinto, R N; de Andrade, A L; Martelli, C M; Zicker, F

1989-01-01

The bacterial flora of 99 cases of abscesses following Bothrops snakebite were analysed. They corresponded to 61.1% of all snakebite abscesses observed in 1030 patients attending the Hospital de Doenças Tropicais de Goiánia in Goiás, Brazil, from January 1984 to April 1988. An exsudate sample of each abscess was examined by Gram stain, culture and susceptibility tests. The Gram negative bacillis, Morganella morganii, Escherichia coli and Providencia sp were the most frequent bacterias isolated. They were identified in 44.4%, 20.2% and 13.1% of the samples respectively. This flora was similar to those described in snake mouth and venom by other researchers. Based on the results of the susceptibility tests the authors suggested the use of chloramphenicol for the treatment of those abscesses which do not respond to simple drainage.

Antimicrobial Susceptibility of Enteric Gram Negative Facultative Anaerobe Bacilli in Aerobic versus Anaerobic Conditions

PubMed Central

Amachawadi, Raghavendra G.; Renter, David G.; Volkova, Victoriya V.

2016-01-01

Antimicrobial treatments result in the host’s enteric bacteria being exposed to the antimicrobials. Pharmacodynamic models can describe how this exposure affects the enteric bacteria and their antimicrobial resistance. The models utilize measurements of bacterial antimicrobial susceptibility traditionally obtained in vitro in aerobic conditions. However, in vivo enteric bacteria are exposed to antimicrobials in anaerobic conditions of the lower intestine. Some of enteric bacteria of food animals are potential foodborne pathogens, e.g., Gram-negative bacilli Escherichia coli and Salmonella enterica. These are facultative anaerobes; their physiology and growth rates change in anaerobic conditions. We hypothesized that their antimicrobial susceptibility also changes, and evaluated differences in the susceptibility in aerobic vs. anaerobic conditions of generic E. coli and Salmonella enterica of diverse serovars isolated from cattle feces. Susceptibility of an isolate was evaluated as its minimum inhibitory concentration (MIC) measured by E-Test® following 24 hours of adaptation to the conditions on Mueller-Hinton agar, and on a more complex tryptic soy agar with 5% sheep blood (BAP) media. We considered all major antimicrobial drug classes used in the U.S. to treat cattle: β-lactams (specifically, ampicillin and ceftriaxone E-Test®), aminoglycosides (gentamicin and kanamycin), fluoroquinolones (enrofloxacin), classical macrolides (erythromycin), azalides (azithromycin), sulfanomides (sulfamethoxazole/trimethoprim), and tetracyclines (tetracycline). Statistical analyses were conducted for the isolates (n≥30) interpreted as susceptible to the antimicrobials based on the clinical breakpoint interpretation for human infection. Bacterial susceptibility to every antimicrobial tested was statistically significantly different in anaerobic vs. aerobic conditions on both media, except for no difference in susceptibility to ceftriaxone on BAP agar. A satellite experiment suggested that during first days in anaerobic conditions the susceptibility changes with time. The results demonstrate that assessing effects of antimicrobial treatments on resistance in the host’s enteric bacteria that are Gram negative facultative Anaerobe Bacilli requires data on the bacterial antimicrobial susceptibility in the conditions resembling those in the intestine. PMID:27191612

Bacteriological profile of neonatal sepsis in a secondary care hospital in rural Tamil Nadu, Southern India.

PubMed

Pavan Kumar, Doniparthi Venkata; Mohan, Jesinth; Rakesh, P S; Prasad, Jasmine; Joseph, Lenikumar

2017-01-01

Neonatal sepsis is a leading cause of neonatal mortality and morbidity in the world. The objective of the current study was to detect the common causative microorganisms of neonatal sepsis and their antimicrobial resistance patterns in a rural secondary hospital in Tamil Nadu, India. Neonates (0-28 days) admitted to this newborn care unit from October 2013 to September 2015, with a diagnosis of probable sepsis were studied. All the enrolled babies had blood cultures taken and were followed up till final outcome, which was discharge or death, irrespective of culture result. Univariate analysis was performed for factors associated with culture positivity, generating odds ratios, and confidence intervals. Among the 107 babies with a diagnosis of probable sepsis, 28 (26.2%) had shown bacteria in culture. The majority (94.4%) were of early-onset sepsis. The predominant organisms were Staphylococcus aureus (10/28) and Klebsiella (6/28). 100% of Gram-negative bacilli and 90% of Staphylococcus were resistant to Ampicillin. Gentamicin resistance among Gram-negative bacilli and Staphylococcus was 52.9% and 20%, respectively, while third-generation cephalosporin resistance was 31.2% and 20%, respectively. Among the neonates diagnosed as probable sepsis, idiopathic prematurity ( P = 0.007) was found to have a statistically significant association with culture-positive sepsis. The culture positivity rate among the neonates with probable sepsis in the current study was 26%. An alarmingly high degree of antibiotic resistance observed calls for robust infection control practices and an urgent evaluation and development of individual and national antibiotic policies for neonatal sepsis.

Beta-lactams in continuous infusion for Gram-negative bacilli osteoarticular infections: an easy method for clinical use.

PubMed

Ribera, Alba; Soldevila, Laura; Rigo-Bonnin, Raul; Tubau, Fe; Padullés, Ariadna; Gómez-Junyent, Joan; Ariza, Javier; Murillo, Oscar

2018-04-01

Continuous infusion (CI) of beta-lactams could optimize their pharmacokinetic/pharmacodynamic indices, especially in difficult-to-treat infections. To validate an easy-to-use method to guide beta-lactams dosage in CI (formula). A retrospective analysis was conducted of a prospectively collected cohort (n = 24 patients) with osteoarticular infections caused by Gram-negative bacilli (GNB) managed with beta-lactams in CI. Beta-lactams dose was calculated using a described formula (daily dose = 24 h × beta-lactam clearance × target "steady-state" concentration) to achieve concentrations above the MIC. We correlated the predicted concentration (C pred  = daily dose/24 h × beta-lactam clearance) with the patient's observed concentration (C obs ) measured by UPLC-MS/MS (Spearman's coefficient). The most frequent microorganism treated was P. aeruginosa (21 cases; 9 MDR). Beta-lactams in CI were ceftazidime (n = 14), aztreonam (7), and piperacillin/tazobactam (3), mainly used in combination (12 with colistin, 5 with ciprofloxacin) and administered without notable side effects. The plasma C obs was higher overall than C pred ; the Spearman correlation between both concentrations was rho = 0.6 (IC 95%: 0.2-0.8) for all beta-lactams, and rho = 0.8 (IC 95%: 0.4-1) for those treated with ceftazidime. The formula may be useful in clinical practice for planning the initial dosage of beta-lactams in CI, while we await a systematic therapeutic drug monitoring. The use of beta-lactams in CI was safe.

Antimicrobial Mechanism of Action of Surfactant Lipid Preparations in Enteric Gram-Negative Bacilli

DTIC Science & Technology

2001-12-25

The addition of EDTA/Tris to the reaction the bacterial cells in the assay time (15min). W60C (1%) mixture results in chelation of ions, facilitating... Maillard , J.Y. (1999) Do antisep- Hamouda, T., Hayes, M.M., Cao, Z. et al. (1999) A novel surfac- tics and disinfectants select for antibiotic resistance

Systemic Edwardsiella tarda infection in a Western African lungfish (Protopterus annectens) with cytologic observation of heterophil projections.

PubMed

Rousselet, Estelle; Stacy, Nicole I; Rotstein, David S; Waltzek, Tom B; Griffin, Matt J; Francis-Floyd, Ruth

2018-06-08

This report describes a case of systemic bacterial infection caused by Edwardsiella tarda in a Western African lungfish (Protopterus annectens) exposed to poor environmental and husbandry conditions. The fish presented with a large, external ulcerative lesion and died 2 weeks after developing anorexia. Histological evaluation revealed multifocal areas of necrosis and heterophilic and histiocytic inflammation throughout multiple tissues. Gram stain identified small numbers of intra- and extracellular monomorphic Gram-negative 1 to 2 μm rod-shaped bacilli. Cytology of lung granuloma, kidney and testes imprints identified heterophilic inflammation with phagocytosis of small monomorphic bacilli and some heterophils exhibiting cytoplasmic projections indicative of heterophil extracellular traps (HETs). Initial phenotypic analysis of isolates from coelomic fluid cultures identified E. tarda. Subsequent molecular analysis of spleen, liver and intestine DNA using an E. tarda-specific endpoint PCR assay targeting the bacterial fimbrial subunit yielded a 115 bp band. Sequencing and BLASTN search revealed the sequence was identical (76/76) to E. tarda strain FL95-01 (GenBank acc. CP011359) and displayed 93% sequence identity (66/71) to Edwardsiella hoshinae strain ATCC 35051 (GenBank acc. CP011359). This is the first report of systemic edwardsiellosis in a lungfish with concurrent cytologically identified structures suggestive of HETs. © 2018 John Wiley & Sons Ltd.

[A case of liver abscess caused by Fusobacterium nucleatum in a patient with recurrent periodontal diseases].

PubMed

Kim, Yong Hwan; Yoon, Hee Jung; Park, Chan Woong; Kim, Jung Ho; Lee, Min Kyung; Kim, Ki Bang; Na, Dong Jib; Kim, Ji Myung

2011-01-01

Fusobacteria are anaerobic gram-negative, non-spore forming bacilli found in normal flora of the oral cavity, urogenital tract, and gastrointestinal tract. Fusobacterium nucleatum has been seldom reported as a cause of liver abscess, particularly in immunocompetent hosts. A 55-year-old man with frequent periodontal disease visited our hospital with intermittent fever and headache for 2 months. Abdominal CT scan revealed an 8.2 × 6 cm mass in the right hepatic lobe with central low density. Abscess culture revealed F. nucleatum as the causative organism. Percutaneous abscess drainage and intravenous administration of antibiotics for 4 weeks improved symptoms and decreased the abscess size. We report a rare case of liver abscess due to F. nucleatum in an immunocompetent man with periodontal disease.

[Epidemiology of nosocomial infections].

PubMed

Astagneau, P

1998-09-15

The frequency of nosocomial infections lies between 5 and 10%, and varies according to the type of hospital and service. Age, underlying disease invasive devices (such as catheters) or procedures are the main risk factors. Common nosocomial infections are urinary tract infections, pneumonia, surgical site infections, bacteremia/septicemia and intravascular catheter-related infections. Gram positive cocci and gram negative bacilli account for one third and two thirds of microorganisms respectively, Staphylococcus aureus being frequently resistant to antibiotics. Prevention is based on a better control of infection risk related to the use of invasive devices.

Hand sanitizer dispensers and associated hospital-acquired infections: friend or fomite?

PubMed

Eiref, Simon D; Leitman, I Michael; Riley, William

2012-06-01

Waterless alcohol-based hand sanitizers are an increasingly popular method of hand hygiene and help prevent hospital-acquired infection (HAI). Whether hand sanitizer dispensers (HSDs) may themselves harbor pathogens or act as fomites has not been reported. All HSDs in the surgical intensive care unit of an urban teaching hospital were cultured at three sites: The dispenser lever, the rear underside, and the area surrounding the dispensing nozzle. All HSDs yielded one or more bacterial species, including commensal skin flora and enteric gram-negative bacilli. Colonization was greatest on the lever, where there is direct hand contact. Hand sanitizer dispensers can become contaminated with pathogens that cause HAI and thus are potential fomites.

Comparison of antral tap with endoscopically directed nasal culture.

PubMed

Casiano, R R; Cohn, S; Villasuso, E; Brown, M; Memari, F; Barquist, E; Namias, N

2001-08-01

The diagnosis of acute bacterial rhinosinusitis continues to generate controversy in critically ill patients. The efficacy of endoscopically directed cultures in these patients is unknown. We compared antral tap (AT) with endoscopic tissue culture (ETC) of the osteomeatal complex in an intensive care unit (ICU) setting. Twenty patients admitted to a surgical/trauma ICU were evaluated by AT and ENB for the presence of rhinosinusitis. All patients had 1) a fever of unknown origin without resolution on empiric antibiotic therapy for > or =48 hrs; 2) other sources of fever ruled out; 3) computed tomography scan evidence of mucoperiosteal thickening +/- sinus air/fluid levels; and 4) attempt at conservative treatment with topical decongestants and removal of all nasal intubation. Microbiologic data were collected and analyzed for any statistical difference between groups. A total of 29 sides underwent simultaneous tap and endoscopically directed tissue culture. The mean age was 40 years (range, 23-77 y) with 85% being males. Fifteen of 20 (75%) patients in the AT group were culture-positive. Of the 49 isolates from the AT, 55% yielded Gram-negative bacilli (Acinetobacter sp. 37%) and 45% yielded Gram-positive cocci. The ETC group was culture-positive in 18 of 20 (90%) patients. Of the 52 isolates from the ETC, Gram-negative bacilli were found in 58% (Acinetobacter sp. 33%) and 42% yielded Gram-positive cocci. The ETCs were culture-positive in all but 1 patient with positive taps. There appeared to be a concordance between AT and ETC in 60% of the patients. In five instances (25%), results of the AT or ETC changed ICU management. Two patients ultimately required sinus surgery. Sinus taps and/or endoscopically directed tissue cultures led to a change in ICU care in 25% of ICU patients studied. In patients with fever of unknown origin and computed tomography evidence of sinusitis, an antral tap continues to provide important information concerning maxillary sinusitis. However, ETC may give as good a representation of the microbiology and secondary inflammatory changes responsible for bacterial ICU rhinosinusitis causing fever of unknown origin. Further study on a larger group of patients is needed.

Antimicrobial susceptibility profiles of gram-negative bacteria causing infections collected across India during 2014-2016: Study for monitoring antimicrobial resistance trend report.

PubMed

Veeraraghavan, Balaji; Jesudason, Mark Ranjan; Prakasah, John Antony Jude; Anandan, Shalini; Sahni, Rani Diana; Pragasam, Agila Kumari; Bakthavatchalam, Yamuna Devi; Selvakumar, Rajesh Joseph; Dhole, T N; Rodrigues, Camilla; Roy, Indranil; Joshi, Sangeetha; Chaudhuri, Bhaskar Narayan; Chitnis, D S

2018-01-01

The emergence of antibiotic resistance among bacterial pathogens in the hospital and community has increased the concern to the health-care providers due to the limited treatment options. Surveillance of antimicrobial resistance (AMR) in frequently isolated bacterial pathogens causing severe infections is of great importance. The data generated will be useful for the clinicians to decide empiric therapy on the local epidemiological resistance profile of the antimicrobial agents. This study aims to monitor the distribution of bacterial pathogen and their susceptibility pattern to the commonly used antimicrobial agents. This study includes Gram-negative bacilli collected from intra-abdominal, urinary tract and respiratory tract infections during 2014-2016. Isolates were collected from seven hospitals across India. All the study isolates were characterised up to species level, and minimum inhibitory concentration was determined for a wide range of antimicrobials included in the study panel. The test results were interpreted as per standard Clinical Laboratory Standards Institute guidelines. A total of 2731 isolates of gram-negative bacteria were tested during study period. The most frequently isolated pathogens were 44% of Escherichia coli (n = 1205) followed by 25% of Klebsiella pneumoniae (n = 676) and 11% of Pseudomonas aeruginosa (n = 308). Among the antimicrobials tested, carbapenems were the most active, followed by amikacin and piperacillin/tazobactam. The rate of extended-spectrum beta-lactamase (ESBL)-positive isolates were ranged from 66%-77% in E. coli to 61%-72% in K. pneumoniae, respectively. Overall, colistin retains its activity in > 90% of the isolates tested and appear promising. Increasing rates of ESBL producers have been noted, which is alarming. Further, carbapenem resistance was also gradually increasing, which needs much attention. Overall, this study data show that carbapenems, amikacin and colistin continue to be the best agents available to treat drug-resistant infections. Thus continuous monitoring of susceptibility profile of the clinically important Gram-negative pathogens is of great importance to guide effective antimicrobial therapy.

Rapid and reliable identification of Gram-negative bacteria and Gram-positive cocci by deposition of bacteria harvested from blood cultures onto the MALDI-TOF plate.

PubMed

Barnini, Simona; Ghelardi, Emilia; Brucculeri, Veronica; Morici, Paola; Lupetti, Antonella

2015-06-18

Rapid identification of the causative agent(s) of bloodstream infections using the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) methodology can lead to increased empirical antimicrobial therapy appropriateness. Herein, we aimed at establishing an easier and simpler method, further referred to as the direct method, using bacteria harvested by serum separator tubes from positive blood cultures and placed onto the polished steel target plate for rapid identification by MALDI-TOF. The results by the direct method were compared with those obtained by MALDI-TOF on bacteria isolated on solid media. Identification of Gram-negative bacilli was 100 % concordant using the direct method or MALDI-TOF on isolated bacteria (96 % with score > 2.0). These two methods were 90 % concordant on Gram-positive cocci (32 % with score > 2.0). Identification by the SepsiTyper method of Gram-positive cocci gave concordant results with MALDI-TOF on isolated bacteria in 87 % of cases (37 % with score > 2.0). The direct method herein developed allows rapid identification (within 30 min) of Gram-negative bacteria and Gram-positive cocci from positive blood cultures and can be used to rapidly report reliable and accurate results, without requiring skilled personnel or the use of expensive kits.

Efficacy of Citrilow® and Cecure® spray wash on the prevalence of aerobic and enterobacteriaceae/gram negative enteric bacilli and cattle hide quality

USDA-ARS?s Scientific Manuscript database

Bacterial cross-contamination from the haired outer surface of cattle to the inner meat as well as hide quality deterioration from bacteria are hygienic problems for meat and hide processing facilities. This study investigated spray wash treatments independently with two antimicrobials used in the ...

Should the patients colonized with extended-spectrum beta-lactamase-producing Gram-negative bacilli (E-GNB) coming to hospital from the community with pneumonia get anti-E-GNB active empirical treatment?

PubMed

Peterlin, Lara; Žagar, Mateja; Lejko Zupanc, Tatjana; Paladin, Marija; Beović, Bojana

2017-10-01

Extended-spectrum beta-lactamases are responsible for resistance of Gram-negative bacilli to several beta-lactam antibiotics, including those prescribed for treatment pneumonia. To evaluate the importance of colonization with E-GNB for the choice of empirical treatment we performed a retrospective case-control study including 156 patients, hospitalized for treatment of pneumonia from 2009 through 2013. Empirical treatment success and in-hospital survival were significantly lower in patients colonized with E-GNB compared to non-colonized (p = 0.002, p = 0.035). When comparing subgroups of colonized patients, treatment success was significantly lower in patients who were colonized with E-GNB resistant to empirical antibiotic (p = 0.010), but not in those colonized by E-GNB susceptible to empirically given antibiotic (p = 0.104). Difference in in-hospital mortality was insignificant in both subgroups (p = 0.056, p = 0.331). The results of study suggest that an anti-E-GNB active antibiotic should be used for empirical treatment of pneumonia in E-GNB colonized patients.

Quinolone-based antibacterial chemoprophylaxis in neutropenic patients: effect of augmented gram-positive activity on infectious morbidity. National Cancer Institute of Canada Clinical Trials Group.

PubMed

Bow, E J; Mandell, L A; Louie, T J; Feld, R; Palmer, M; Zee, B; Pater, J

1996-08-01

To determine whether augmented quinolone-based antibacterial prophylaxis in neutropenic patients with cancer reduces infections caused by gram-positive cocci and preserves the protective effect against aerobic gram-negative bacilli. Open, randomized, controlled, multicenter clinical trial. Centers participating in the National Cancer Institute of Canada Clinical Trials Group. 111 eligible and evaluable patients hospitalized for severe neutropenia (neutrophil count < 0.5 x 10(9)/L lasting at least 14 days) who were receiving cytotoxic therapy for acute leukemia or bone marrow autografting. One of three oral antibacterial prophylactic regimens (norfloxacin, 400 mg every 12 hours; ofloxacin, 400 mg every 12 hours; or ofloxacin, 400 mg, plus rifampin, 300 mg every 12 hours) beginning with cytotoxic therapy. Incidence and cause of suspected or proven infection. Microbiologically documented overall infection rates for norfloxacin, ofloxacin, and ofloxacin plus rifampin were 47%, 24%, and 9%, respectively (P < 0.001). Corresponding rates were 24%, 13%, and 3%, respectively for staphylococcal bacteremia (P = 0.03) and, 21%, 3%, and 3%, respectively for streptococcal bacteremia (P < 0.01). The pattern of bacteremia suggested that rifampin played a role in suppressing staphylococcal infection. Both ofloxacin alone and ofloxacin plus rifampin had a clinically significant antistreptococcal effect. Aerobic gram-negative rods were cleared from rectal surveillance cultures in all patients after a median of 5.5 days and caused infection in only one patient (0.9%). The reductions in the number of microbiologically documented infections among ofloxacin recipients and ofloxacin plus rifampin recipients were offset by concomitant increases in the number of unexplained fevers (24% of norfloxacin recipients, 53% of ofloxacin recipients, and 49% of ofloxacin plus rifampin recipients; P = 0.02). No statistically significant difference was found among the treatment arms with respect to the overall incidence of febrile neutropenic episodes as defined for this trial (79% for the norfloxacin group, 82% for the ofloxacin group, and 77% for the ofloxacin plus rifampin group). Quinolone-based antibacterial chemoprophylaxis protected patients from aerobic gram-negative bacillary infections. Augmentation of the gram-positive activity reduced the incidence of gram-positive infections but did not influence the overall incidence of febrile neutropenic episodes.

Identification of aerobic Gram-positive bacilli by use of Vitek MS.

PubMed

Navas, Maria; Pincus, David H; Wilkey, Kathy; Sercia, Linda; LaSalvia, Margaret; Wilson, Deborah; Procop, Gary W; Richter, Sandra S

2014-04-01

The accuracy of Vitek MS mass spectrometric identifications was assessed for 206 clinically significant isolates of aerobic Gram-positive bacilli representing 20 genera and 38 species. The Vitek MS identifications were correct for 85% of the isolates (56.3% to the species level, 28.6% limited to the genus level), with misidentifications occurring for 7.3% of the isolates.

Evolution of sensitivity to fosfomycin in bacteria isolated in 1973, 1974 and 1975 in the Servicio de Microbiologia y Epidemiologia of the 'Clinica Puerta de Hierro', Madrid.

PubMed

Dámaso, D; Moreno-López, M; Martínez-Beltrán, J

1977-01-01

The bacteriostatic activity of fosfomycin was studied in vitro against 1,243 clinical isolations of gram-positive cocci and 4,086 isolations of gram-negative bacilli that were obtained in 1973, 1974 and in the period from January to May of 1975. MIC was determined by the agar diffusion method, quantifying it by means of the standard curve that was worked out with the strain of E. coli NCTC 10,418. A slight increase in resistance was observed in the gram-positive cocci: 64 mug/ml were inhibitory for 63% of the 249 isolations obtained in 1973, 59.1% of the 716 isolations obtained in 1974, and 57.5% of the 278 isolations from 1975. A slight loss of sensitivity was also observed in the gram-negative bacilli: the aforementioned concentration of fosfomycin inhibited 36% of the 742 isolations from 1973, 33.6% of the 2,387 isolations from 1974 and 32.6% of the 957 isolations from 1975. 933 g of this antibiotic were consumed in our hospital in 1973, 4,203 g in 1974 and 957 g in 1975. The consumption rate per patient per year was 0.15, 0.72 and 0.20 g, respectively. In conclusion, although no change was observed in the sensitivity of some bacterial strains to fosfomycin, the overall study indicates a slight decrease in the sensitivity, although it does not apparently have any relationship to the consumption of fosfomycin in our hospital.

ANTIBIOTIC RESISTANCE SPECTRUM OF NON FERMENTING GRAM NEGATIVE BACILLI ISOLATED IN THE ORTHOPEDIC TRAUMATOLOGY CLINIC OF "SF. SPIRIDON" CLINICAL EMERGENCY HOSPITAL IAŞI.

PubMed

Tucaliuc, D; Alexa, O; Tuchiluş, Cristina Gabriela; Ursu, Ramona Gabriela; Tucaliuc, Elena Simona; Jelihovsky, I; Iancu, Luminiţa Smaranda

2015-01-01

The retro-prospective analysis of antibiotic sensitivity of non-fermenting gram negative bacilli strains circulating in the Orthopedics-Traumatology Clinic from "Sf. Spiridon" Emergency Clinical Hospital in view of determining the trend of the resistance phenomenon and indicating the most useful treatment for the infections caused by these strains. The retrospective component was conducted from 01.01.2003 to 31.12.2012, and the result of the diffusimetric antibiograms was taken from the hospital's informatics system; the prospective component of the study involved the collection of pathological products from the patients admitted during January-December 2013, who showed clinical suspicion of infection, in compliance with the general collection norms for the products destined for the bacteriological exam. From the total 167 strains of Pseudomonas aeruginosa isolated and identified from the patients, 48 (28.74%) were sensitive to at least one antibiotic from each tested class, 29 (17.39%) were resistant to a single antibiotic and the rest of 90 (53.89%) showed multiple resistance. We noticed a statistically significant difference between the number of strains sensitive to at least one antibiotic from each tested class and those with multiple resistance (p < 0.05). For the strains of Acinetobacter baumanii combined resistance was identified for 121 (87.04%), out of which 55 (39.56%) were resistant to two classes of antibiotics and the other (47.48%) to all three classes. The most frequently met was the association of resistance to quinolones and aminoglycosides, namely for a number of 49 strains (35.25%); only 3.59% of them were simultaneously sensitive to the three classes of antibiotics. The already high percentages and the rising trends of antibiotic resistance of non-fermenting gram-negative bacteria described in this study confirm the continuous decrease of the efficiency of antimicrobial agents and underline the necessity of a global strategy which aims at all health sectors regarding the rational use of antibiotics, on the one hand, and the continuation of studies concerning the surveillance of the antimicrobial resistance phenomenon, on the other hand.

Clinical pulmonary infection score and a spot serum procalcitonin level to guide discontinuation of antibiotics in ventilator-associated pneumonia: a study in a single institution with high prevalence of nonfermentative gram-negative bacilli infection.

PubMed

Wongsurakiat, Phunsup; Tulatamakit, Sirapat

2018-01-01

Background We wanted to determine the impact of combined Clinical Pulmonary Infection Score (CPIS) and a spot serum procalcitonin (PCT)-guided protocol to shorten the duration of antibiotic treatment in patients with ventilator-associated pneumonia (VAP), mainly caused by nonfermentative gram-negative bacilli (NF-GNB). Methods Patients with VAP who received appropriate antibiotics for 7 days, temperature ⩽ 37.8°C, without shock, and CPIS ⩽ 6 were allocated to the PCT group or conventional group according to the treating physicians' decisions. In the PCT group, antibiotics were stopped if the PCT level on day 8 < 0.5 ng/ml. In the conventional group, antibiotics were stopped according to physicians' discretion. Results There were 24 patients in the PCT group and 26 patients in the conventional group. NF-GNB were responsible for VAP in 79.2% of the PCT group and 65.4% of the conventional group. PCT group had a greater number of antibiotic-free days alive during the 28 days after VAP onset than the conventional group (14.6 ± 5.4 days versus 5.9 ± 5.7 days, respectively; p <.001). In the multivariate, propensity score-adjusted analysis, the PCT group [coefficient = -9.1 (-12.2 to -6); p <.001] and extrapulmonary infections [coefficient = 6.4 (3.3-9.5); p <.001] were independent predictors of total antibiotic exposure days. There was no relapse in both groups. Meanwhile, 12.5% of the PCT group and 26.9% of the conventional group subsequently developed recurrent VAP compatible with superinfections. Conclusions CPIS and a spot serum PCT level appeared effective and safe to guide discontinuation of antibiotic treatment in patients with VAP caused by NF-GNB. TCTR20160726002.

Etiology of Bacteremia in Young Infants in Six Countries

PubMed Central

Darmstadt, Gary L.; Carlin, John B.; Zaidi, Anita K. M.; Yeboah-Antwi, Kojo; Saha, Samir K.; Ray, Pallab; Narang, Anil; Mazzi, Eduardo; Kumar, Praveen; Kapil, Arti; Jeena, Prakash M.; Deorari, Ashok; Chowdury, A.K. Azad; Bartos, Andrés; Bhutta, Zulfiqar A.; Adu-Sarkodie, Yaw; Adhikari, Miriam; Addo-Yobo, Emmanuel; Weber, Martin W.

2015-01-01

Background: Neonatal illness is a leading cause of death worldwide; sepsis is one of the main contributors. The etiologies of community-acquired neonatal bacteremia in developing countries have not been well characterized. Methods: Infants <2 months of age brought with illness to selected health facilities in Bangladesh, Bolivia, Ghana, India, Pakistan and South Africa were evaluated, and blood cultures taken if they were considered ill enough to be admitted to hospital. Organisms were isolated using standard culture techniques. Results: Eight thousand eight hundred and eighty-nine infants were recruited, including 3177 0–6 days of age and 5712 7–59 days of age; 10.7% (947/8889) had a blood culture performed. Of those requiring hospital management, 782 (54%) had blood cultures performed. Probable or definite pathogens were identified in 10.6% including 10.4% of newborns 0–6 days of age (44/424) and 10.9% of infants 7–59 days of age (39/358). Staphylococcus aureus was the most commonly isolated species (36/83, 43.4%) followed by various species of Gram-negative bacilli (39/83, 46.9%; Acinetobacter spp., Escherichia coli and Klebsiella spp. were the most common organisms). Resistance to second and third generation cephalosporins was present in more than half of isolates and 44% of the Gram-negative isolates were gentamicin-resistant. Mortality rates were similar in hospitalized infants with positive (5/71, 7.0%) and negative blood cultures (42/557, 7.5%). Conclusions: This large study of young infants aged 0–59 days demonstrated a broad array of Gram-positive and Gram-negative pathogens responsible for community-acquired bacteremia and substantial levels of antimicrobial resistance. The role of S. aureus as a pathogen is unclear and merits further investigation. PMID:25389919

Etiology of bacteremia in young infants in six countries.

PubMed

Hamer, Davidson H; Darmstadt, Gary L; Carlin, John B; Zaidi, Anita K M; Yeboah-Antwi, Kojo; Saha, Samir K; Ray, Pallab; Narang, Anil; Mazzi, Eduardo; Kumar, Praveen; Kapil, Arti; Jeena, Prakash M; Deorari, Ashok; Chowdury, A K Azad; Bartos, Andrés; Bhutta, Zulfiqar A; Adu-Sarkodie, Yaw; Adhikari, Miriam; Addo-Yobo, Emmanuel; Weber, Martin W

2015-01-01

Neonatal illness is a leading cause of death worldwide; sepsis is one of the main contributors. The etiologies of community-acquired neonatal bacteremia in developing countries have not been well characterized. Infants <2 months of age brought with illness to selected health facilities in Bangladesh, Bolivia, Ghana, India, Pakistan and South Africa were evaluated, and blood cultures taken if they were considered ill enough to be admitted to hospital. Organisms were isolated using standard culture techniques. Eight thousand eight hundred and eighty-nine infants were recruited, including 3177 0-6 days of age and 5712 7-59 days of age; 10.7% (947/8889) had a blood culture performed. Of those requiring hospital management, 782 (54%) had blood cultures performed. Probable or definite pathogens were identified in 10.6% including 10.4% of newborns 0-6 days of age (44/424) and 10.9% of infants 7-59 days of age (39/358). Staphylococcus aureus was the most commonly isolated species (36/83, 43.4%) followed by various species of Gram-negative bacilli (39/83, 46.9%; Acinetobacter spp., Escherichia coli and Klebsiella spp. were the most common organisms). Resistance to second and third generation cephalosporins was present in more than half of isolates and 44% of the Gram-negative isolates were gentamicin-resistant. Mortality rates were similar in hospitalized infants with positive (5/71, 7.0%) and negative blood cultures (42/557, 7.5%). This large study of young infants aged 0-59 days demonstrated a broad array of Gram-positive and Gram-negative pathogens responsible for community-acquired bacteremia and substantial levels of antimicrobial resistance. The role of S. aureus as a pathogen is unclear and merits further investigation.

[Clinical analysis for patients with continuous ambulatory peritoneal dialysis associated peritonitis].

PubMed

Liu, Jian; Huang, Xun; Liu, Yao; Xu, Hui; Gong, Rui'e; Li, Chunhui

2016-12-28

To analyze the clinical characteristics of continuous ambulatory peritoneal dialysis (CAPD) associated peritonitis in the tertiary hospitals and to discuss the preventive and therapeutic strategy.
 Methods: The clinical characteristics, pathogens, resistance and outcomes of 126 CAPD associated peritonitis in 104 patients from Jan, 2013 to June, 2016, were retrospectively analyzed.
 Results: Among the patients, the incidence rates of abdominal pain, fever, diarrhea and emesis were 104 (82.54%), 56 (44.44%), 49 (38.89%), and 31 (23.60%), respectively. Among them, 88 patients suffered peritonitis once, other 16 patients suffered multiple peritonitis or recurrent peritonitis for 38 times. Among the 38 times, the numbers for recurrent, repeated or catheter-associated peritonitis were 2, 2, or 3, respectively. Peritoneal fluids from 103 cases were cultured, and 64 cases were positive in bacteria, with a rate of 62.14%. A total of 70 strains of bacteria were separated, including 42 strains of gram-positive bacteria, 21 strains of gram-negative bacteria, and 7 strains of fungus. The most common gram-positive pathogens were Staphylococcus epidermidis, Enterococcus faecalis and Staphylococcus haemolyticus, while Escherichia coli, Klebsiella pneumoniae and Klebsiella pneumoniae were the most common gram-negative bacteria. Candida albicans was the major fungal pathogens. Gram-positive cocci showed resistance to gentamycin, levofloxacin, moxifloxacin, vancomycin and linezolid, with a rate at 20.00%, 36.11%, 5%, 0%, and 0%, respectively. The gram-negative bacilli were resistent to cefoperazone/sulbactam, gentamycin, cephazolin, and ceftazidime, with a rate at 6.25%, 10.53%, 64.29%, and 15.38%, respectively. There were no imipenem, amikacin, piperacillin/tazobactam-resistant strains were found.
 Conclusion: The most common pathogen causing CAPD associated peritonitis is gram-positive bacteria. It is crucial to take the anti-infection therapy for CAPD associated peritonitis early. The positive rates for bacterial culture need to be enhanced through improvement of methods. At the same time, doctors could improve the outcome of CAPD associated peritonitis by adjusting the medication according to the drug sensitivity results.

Staphylococcal scalded skin syndrome in a premature newborn caused by methicillin-resistant Staphylococcus aureus: case report.

PubMed

Hörner, Andreas; Hörner, Rosmari; Salla, Adenilde; Nunes, Melise Silveira; Garzon, Litiérri Razia; Rampelotto, Roberta Filipini; Martini, Rosiéli; Santos, Silvana Oliveira dos; Gindri, Lívia; Rodrigues, Mônica de Abreu; Giacomolli, Cláudia

2015-01-01

Staphylococcal scalded skin syndrome is an exfoliative skin disease. Reports of this syndrome in newborns caused by methicillin-resistant Staphylococcus aureus are rare but, when present, rapid diagnosis and treatment is required in order to decrease morbidity and mortality. A premature newly born girl weighing 1,520 g, born with a gestational age of 29 weeks and 4 days, developed staphylococcal scalded skin syndrome on the fifth day of life. Cultures on blood samples collected on the first and fourth days were negative, but Pseudomonas aeruginosa and Enterococcus sp. (vancomycin-sensitive) developed in blood cultures performed on the day of death (seventh day), and Pseudomonas aeruginosa and Serratia marcescens were identified in cultures on nasopharyngeal, buttock and abdominal secretions. In addition to these two Gram-negative bacilli, methicillin-resistant Staphylococcus aureus was isolated in a culture on the umbilical stump (seventh day). The diagnosis of staphylococcal scalded skin syndrome was based on clinical criteria.

Identification of Aerobic Gram-Positive Bacilli by Use of Vitek MS

PubMed Central

Navas, Maria; Pincus, David H.; Wilkey, Kathy; Sercia, Linda; LaSalvia, Margaret; Wilson, Deborah; Procop, Gary W.

2014-01-01

The accuracy of Vitek MS mass spectrometric identifications was assessed for 206 clinically significant isolates of aerobic Gram-positive bacilli representing 20 genera and 38 species. The Vitek MS identifications were correct for 85% of the isolates (56.3% to the species level, 28.6% limited to the genus level), with misidentifications occurring for 7.3% of the isolates. PMID:24501030

Synthesis and Antimicrobial Activity of Some Novel Cross-Linked Chitosan Hydrogels

PubMed Central

Mohamed, Nadia Ahmed; Fahmy, Mona Mohamed

2012-01-01

Four novel hydrogels based on chitosan were synthesized via a cross-linking reaction of chitosan with different concentrations of oxalyl bis 4-(2,5-dioxo-2H-pyrrol- 1(5H)-yl)benzamide. Their structures were confirmed by fourier transform infrared X-ray (FTIR), scanning electron microscopy (SEM) and X-ray diffraction. The antimicrobial activities of the hydrogels against two crop-threatening pathogenic fungi namely: Aspergillus fumigatus (A. fumigatus, RCMBA 06002), and Aspergillus niger (A. niger, RCMBA 06106), and five bacterial species namely: Bacillis subtilis (B. subtilis, RCMBA 6005), Staphylococcus aureus (S. aureus, RCMBA 2004), Streptococcus pneumoniae (S. pneumonia, RCMB 000101) as Gram positive bacteria, and Salmonella typhimurium (S. typhimurium, RCMB 000104), and Escherichia coli (E. coli, RCMBA 5003) as Gram negative bacteria have been investigated. The prepared hydrogels showed much higher antimicrobial activities than that of the parent chitosan. The hydrogels were more potent in case of Gram-positive bacteria than Gram-negative bacteria. Increasing the degree of cross-linking in the hydrogels resulted in a weaker antimicrobial activity. PMID:23109847

Infectious peritonitis profile in peritoneal dialysis at Ibn Sina University Hospital: a 6-year data report.

PubMed

Bekaoui, Samira; Haddiya, Intissar; Houti, Maria Slimani; Berkchi, Fatima Zahra; Ezaitouni, Fatima; Ouzeddoun, Naima; Bayahia, Rabia; Benamar, Loubna

2014-01-01

Infectious peritonitis (IP) is the most common complication in peritoneal dialysis (PD). The purpose of this study is to assess the prevalence of IP and to determine its clinical, biological, and evolutive characteristics. We conducted a five year, five months retrospective study from July 2006 to December 2011. All patients on peritoneal dialysis that have been followed on PD for a minimum of 3 months and who presented IP during follow-up were included. Data were analyzed using SPSS 17.0. The 76 episodes of IP were identified in 36 patients. The peritonitis rate (months × patients/peritonitis), as calculated by the Registre de Dialyse Péritonéale de Langue Française (RDPLF Registry) [French peritoneal dialysis registry] in December 2011, was 18.59. Time to occurrence of peritonitis from the start of peritoneal exchange was 15.44±10 months. The mean age of our patients was 49.1±16.8 years [10-80]: the youngest patient's age was 10, while the oldest was 80 years old (male to female: sex ratio M/F=1,66). Also, 22% of our patients were diabetic. The mean follow-up in PD was 22.6±14 months. Abdominal pain was present in 79% of the cases. Fever and vomiting were noted in 42% and 38% of cases, respectively. The C-reactive protein rate was elevated in 77% of cases, and leukocytosis was found in 27% of cases. Bacteriological proof was present in 73.68% of cases. Gram-positive cocci were involved in 56.6% of microbiologically proven IP cases. Gram-negative bacilli were represented in 37.7%. The outcome was favorable in 89.4%. The PD catheter was removed in 2.63% of the cases. In addition, 7.89% of our patients were transferred to hemodialysis. The rate of IP remains high in our series. More than one-half of the peritonitis cases with positive cultures (56.6%) were caused by Gram-positive cocci. Gram-negative bacilli ranked second (27.7%). These results agree with data in the literature. Moreover, the rate of culture-negative IP in our series is high (26%). Evolution is good in most cases (89%). Despite the gradual decrease of its rate, peritonitis remains frequent in our center and calls for optimization of means of prevention. The high frequency of negative culture IP in our study urges us toward better collaboration with biologists to target antibiotic therapy and improve IP management.

Polymicrobial tenosynovitis with Pasteurella multocida and other Gram negative bacilli after a Siberian tiger bite

PubMed Central

Isotalo, P; Edgar, D; Toye, B

2000-01-01

Mammalian bites present a considerable clinical problem because they are often associated with bacterial infections. Pasteurella multocida is a microorganism that commonly infects both canine and small feline bites. Zoonotic infections developing after large feline bites have been recognised, although their reports are limited. We describe a 35 year old man who was bitten by a Siberian tiger and who developed infectious tenosynovitis secondary to P multocida, Bergeyella (Weeksella) zoohelcum, and Gram negative bacteria most like CDC group EF-4b and comamonas species. The latter three bacteria have not been isolated previously from large feline bite wounds. Key Words: animal bite • zoonotic infection • Pasteurella multocida • Bergeyella zoohelcum PMID:11127273

Automated reading of MIC microdilution trays containing fluorogenic enzyme substrates with the Sensititre Autoreader.

PubMed Central

Staneck, J L; Allen, S D; Harris, E E; Tilton, R C

1985-01-01

The Sensititre Autoreader is a microcomputer-driven instrument capable of automatically reading antimicrobial susceptibility microdilution trays. The instrument measures the fluorescence liberated by bacterial enzymatic activity on fluorogenic substrates as an indicator of growth in each well. A mathematical algorithm converts the fluorescent signals from an antimicrobial dilution series to an MIC endpoint. A three-center study evaluated the performance of the Autoreader in comparison with MIC determined visually in a duplicate set of control plates lacking fluorogenic substrate. Among 828 isolates of gram-negative bacilli tested against 17 antimicrobial agents, Autoreader 18-h MIC were within +/- 1 twofold dilution of control MIC values (agreement) in 95.3% of instances. In 3.5% of the instances, Autoreader values occurred +/- 2 half-step dilutions from control values (minor discrepancy), and in only 1.2% of instances did Autoreader values deviate from control values by greater than +/- 2 dilution steps (major discrepancy). Agreement, minor discrepancies, and major discrepancies were noted among 148 gram-positive cocci tested against 11 antimicrobial agents in 93.5, 4.8, and 1.7% of the instances, respectively. Over half of the major discrepancies noted with gram-negative bacilli occurred with Proteus mirabilis-beta-lactam combinations, a problem that was resolved when a lower initial inoculum was used. Inter-and intralaboratory reproducibility was excellent. Standard Sensititre susceptibility trays may be instrument read at 18 h reproducibly and accurately with only slight modification of conventional procedures to include fluorogenic enzyme substrates in the incubation broth. PMID:4031033

Gram stain microbiological pattern of upper extremities suppuration at Baptist Medical Centre, Ogbomoso Nigeria: a fifteen month review.

PubMed

Oke, A J; Olaolorun, D A; Meier, D E; Tarpley, J L

2011-06-01

Sixty-eight (68) patients with serious upper extremity suppurative infections, presenting within a period of fifteen (15) months, were prospectively studied clinically, Gram stain of aspirates/pus were performed, specimen cultured, planted, and where indicated glucose levels and haemoglobin genotype determined. Half of the patients had hand infections. Staphylococcus aureus was isolated from thirty-nine (39) patients. Gram Negative bacilli, including Salmonella were more isolated from patients with diabetes mellitus or Hgb SS or SC. The Gram stain results correlated with the culture result 90%. When Gram Positive cocci were demonstrated in the primary microscopic examination, cultures were not mandatory. When no organism was demonstrated on primary Gram stain or the patient was diabetic or a sickler, cultures of the specimens were done. The Gram stain, well performed, remains a useful, inexpensive, technologically appropriate laboratory test for abetting decision making in patients with upper extremity suppurative infections. Organisms encountered in this study included: Staphylococcus aureus, Streptococcus pyogenes, Salmonella typhi, Proteus mirabilis, Pseudomonas aeruginosa, and Coliforms.

β-Lactamase Production in Key Gram-Negative Pathogen Isolates from the Arabian Peninsula

PubMed Central

Balkhy, Hanan H.; Walsh, Timothy R.; Paterson, David L.

2013-01-01

SUMMARY Infections due to Gram-negative bacilli (GNB) are a leading cause of morbidity and mortality worldwide. The extent of antibiotic resistance in GNB in countries of the Gulf Cooperation Council (GCC), namely, Saudi Arabia, United Arab Emirates, Kuwait, Qatar, Oman, and Bahrain, has not been previously reviewed. These countries share a high prevalence of extended-spectrum-β-lactamase (ESBL)- and carbapenemase-producing GNB, most of which are associated with nosocomial infections. Well-known and widespread β-lactamases genes (such as those for CTX-M-15, OXA-48, and NDM-1) have found their way into isolates from the GCC states. However, less common and unique enzymes have also been identified. These include PER-7, GES-11, and PME-1. Several potential risk factors unique to the GCC states may have contributed to the emergence and spread of β-lactamases, including the unnecessary use of antibiotics and the large population of migrant workers, particularly from the Indian subcontinent. It is clear that active surveillance of antimicrobial resistance in the GCC states is urgently needed to address regional interventions that can contain the antimicrobial resistance issue. PMID:23824364

Pseudoclavibacter otitis media in a 3-year-old boy with pulmonary and spinal tuberculosis.

PubMed

Ayuthaya, Satja Issaranggoon na; Leelaporn, Amornrut; Kiratisin, Pattarachai; Oberdorfer, Peninnah

2015-05-01

Pseudoclavibacter has rarely been documented as an etiologic agent of infection in humans. We presented the first case report of Pseudoclavibacter otitis media in a boy with pulmonary and spinal tuberculosis.A 3-year-old boy was referred to our hospital due to prolonged fever and progressive paraplegia for 3 months. He had yellowish discharge from both ear canals. The pleural fluid culture was positive for Mycobacterium tuberculosis. The discharge from both ears culture yielded yellow colonies of gram-positive bacilli with branching. This organism was positive for modified acid-fast bacilli stain but negative for acid-fast bacilli stain. Biochemical characteristics of this isolate were positive for catalase test but negative for oxidase, nitrate, esculin, and sugar utilization tests. The organism was further subjected to be identified by 16S ribosomal deoxyribonucleic acid gene sequencing. The result yielded Pseudoclavibacter species (99.4% identical), which could be most likely a potential pathogen in immunocompromised host like this patient. He responded well with intravenous trimetroprim-sulfamethoxazole for 6 weeks.This is the first case report of Pseudoclavibacter otitis media in children, and this case could emphasize Pseudoclavibacter species as a potential pathogen in immunocompromised host.

Pseudoclavibacter Otitis Media in a 3-Year-Old Boy With Pulmonary and Spinal Tuberculosis

PubMed Central

Ayuthaya, Satja Issaranggoon na; Leelaporn, Amornrut; Kiratisin, Pattarachai; Oberdorfer, Peninnah

2015-01-01

Abstract Pseudoclavibacter has rarely been documented as an etiologic agent of infection in humans. We presented the first case report of Pseudoclavibacter otitis media in a boy with pulmonary and spinal tuberculosis. A 3-year-old boy was referred to our hospital due to prolonged fever and progressive paraplegia for 3 months. He had yellowish discharge from both ear canals. The pleural fluid culture was positive for Mycobacterium tuberculosis. The discharge from both ears culture yielded yellow colonies of gram-positive bacilli with branching. This organism was positive for modified acid-fast bacilli stain but negative for acid-fast bacilli stain. Biochemical characteristics of this isolate were positive for catalase test but negative for oxidase, nitrate, esculin, and sugar utilization tests. The organism was further subjected to be identified by 16S ribosomal deoxyribonucleic acid gene sequencing. The result yielded Pseudoclavibacter species (99.4% identical), which could be most likely a potential pathogen in immunocompromised host like this patient. He responded well with intravenous trimetroprim-sulfamethoxazole for 6 weeks. This is the first case report of Pseudoclavibacter otitis media in children, and this case could emphasize Pseudoclavibacter species as a potential pathogen in immunocompromised host. PMID:25929901

Rapid detection of multidrug resistant Gram-negative bacilli by Cica-Beta-Test strips.

PubMed

Lavigne, J-P; Pfeiffer, C; Vidal, L; Sotto, A

2011-02-01

This study aimed to evaluate the sensitivity and specificity of a new system (Cica-Beta-Test, Kanto Chemical, Japan) for rapid detection of AmpC-derepressed, extended-spectrum β-lactamases (ESβL) and metallo-β-lactamases (MβL). Two hundred Multi-Drug Resistant (MDR)-Gram-negative bacilli were studied: 170 Enterobacteriaceae and 30 Gram-negative non-fermentative bacteria. One hundred and eighteen strains produced an ESβL, seven MβL and 75 derepressed cephalosporinases. One drop of substrate was dispensed onto the filter pad of the Cica-Beta-Test strip. The bacterial colonies were spread on the filter pad of strip. The reading was performed after 2 to 15 min: turning chromogenic indicated the positive test. Three tests were used: Cica-Beta I for detection of MDR bacteria; Cica-Beta MβL for detection of MβL-producing bacteria and Cica-Beta CVA, which distinguish ESβL and AmpC-derepressed producers. Results were compared with molecular assays. Cica-Beta-Test I has detected 194 MDR (sensitivity 97%), Cica-Beta-Test MβL has shown the presence of six MβL tested (sensitivity 85.7%). Five strains were non-MβL false positive (specificity 97.3%). Cica-Beta-Test CVA allowed the differentiation of ESβL-producing strains (109/115) and AmpC-derepressed strains (56/67) (sensitivity 94.8%, specificity 83.8%). Because of their epidemic nature, the MDR strains are screened and require strict hygienic measures patients. The simultaneous use of three strips can quickly determine the presence of MDR including ESβL and MβL. Rapid screening of MDR avoids transmission and limits the use of broad-spectrum antibiotics. Copyright © 2010 Elsevier Masson SAS. All rights reserved.

Risk factors associated with fluoroquinolone-resistant enterococcal urinary tract infections in a tertiary care university hospital in north India.

PubMed

Banerjee, Tuhina; Anupurba, Shampa

2016-10-01

Fluoroquinolone resistance in both Gram-positive and Gram-negative bacteria has increased with the widespread use of fluoroquinolones. Fluoroquinolone resistance in Gram-negative bacilli has been widely studied, though staphylococci and enterococci are also notably resistant. Enterococci being the second most common cause of healthcare-associated urinary tract infections (UTIs) fluoroquinolones are often the drug of choice. This study was undertaken to assess the risk factors associated with fluoroquinolone-resistant enterococcal UTI in a tertiary level health facility in north India. A total of 365 patients with UTI caused by enterococci were studied over a period of two years. Patients with ciprofloxacin-resistant and susceptible UTI were considered as cases and controls, respectively. Resistance profile of the isolates against common antibiotics was studied by minimum inhibitory concentration (MIC) determination. Mechanisms for fluoroquinolone resistance was studied by efflux pump inhibitor activity and multiplex PCR targeting the qnr genes. A total of 204 (55.89%) cases and 161 (44.1%) controls were identified. The fluoroquinolone-resistant isolates were significantly resistant to ampicillin, high strength aminoglycosides and vancomycin. The majority (78%) of the resistant isolates showed efflux pump activity. Treatment in indoor locations, presence of urinary catheters and pregnancy along with recent exposure to antibiotics especially fluoroquinolones, third generation cephalosporins and piperacillin-tazobactam were identified as independent risk factors. Our results showed that fluoroquinolone resistance in enterococcal UTI was largely associated with indoor usage of antibiotics and use of indwelling devices. Knowledge of risk factors is important to curb this emergence of resistance.

Microbiology, infection control and infection related outcome in pediatric patients in an oncology center in Eastern India: Experience from Tata Medical Center, Kolkata.

PubMed

Bhattacharyya, Arpita; Krishnan, Shekhar; Saha, Vaskar; Goel, Gaurav; Bhattacharya, Sanjay; Hmar, Lalawmpuia

2014-01-01

Infection is a major determinant in the outcome of patients with cancer. The aim was to know the epidemiology and outcome of patients with cancer in a cancer care center in Eastern India. Retrospective study of pediatric patients in Tata Medical Center, Kolkata, India. Patients (n = 262) between the age group of 0 and 18 years were reviewed for infections and infection-related outcome (January to December 2013). Modified Wald method was used to determine confidence interval of proportions. Gram-negative bacteria were found to be the most common cause of bloodstream infections (BSIs) (56.4%), followed by Gram-positive cocci (34.5%), and Candida species (9.1%). Carbapenem-resistance was noted among 24% of Gram-negative bacilli (GNB), and extended-spectrum beta-lactamase among 64% of GNBs. A single case of Vibrio cholerae septicemia was also noted. No case of vancomycin-resistant Enterococcus was observed, whereas only two cases of methicillin-resistant Staphylococcus aureus bacteremia (1/3 of all Staphylococcus aureus bacteremia) were detected. Escherichia coli, followed by Klebsiella, Pseudomonas, and Acinetobacter were the predominant organisms detected in BSIs. Among Candida spp. BSIs no resistance to caspofungin, amphotericin B, Voriconazole was noted. Candida tropicalis was the most common isolate, and 1 isolate of Candida glabrata showed dose-dependent sensitivity to fluconazole. Three out of 25 patients died of multi-drug resistant Gram-negative bacteria (12%) in 2013. Seventeen patients had radiological evidence of invasive fungal infections (no mortality was noted). Periodic review of infection-related data, as well as infection control practices, is essential to optimize clinical outcome in patients with pediatric malignancies.

Severe Legionnaire's disease caused by Legionella longbeachae in a long-term renal transplant patient: the importance of safe living strategies after transplantation.

PubMed

Wright, A J; Humar, A; Gourishankar, S; Bernard, K; Kumar, D

2012-08-01

Legionella species are intracellular gram-negative bacilli that require specific culture media for growth. Transplant recipients with impaired cellular immunity are at particular risk for infection with this pathogen. Most human disease is caused by Legionella pneumophila; disease caused by non-L. pneumophila species is reported mainly in immunosuppressed patients with the exception of Legionella longbeachae. L. longbeachae is a common cause of Legionnaires' disease in Australia and New Zealand, and is associated with exposure to potting soil. We report the case of a patient, 26 years post kidney transplant, who presented with severe and rapidly progressive respiratory illness. L. longbeachae serogroup 1 was isolated from respiratory cultures. Further investigation revealed that she had significant soil exposure before the onset of illness. We highlight the importance of following safe living strategies to prevent exposure-related illness even in long-term transplant recipients. © 2012 John Wiley & Sons A/S.

Bovine esophageal and glossal ulceration associated with Pseudomonas aeruginosa and Fusobacterium spp.in a 10-month-old Holstein heifer.

PubMed

Tosaki, Kaori; Kojima, Hirokazu; Akama, Shunsuke; Ootake, Yoshihiro; Inoue, Kyoichi; Katsuda, Ken; Shibahara, Tomoyuki

2018-05-25

An underweight 10-month-old Holstein heifer presented with anorexia and ananastasia and was euthanized. Postmortem examination revealed extensive ulceration in the esophagus, tongue, and omasum. Histopathological examination revealed severe necrotic esophagitis, glossitis, and omasitis. Many Gram-negative bacilli were detected throughout the necrotic area in the digestive tract; these were identified as Pseudomonas aeruginosa on the basis of isolation tests, molecular examinations, and immunohistochemistry. Gram-negative long filamentous organisms in the superficial layers of the necrotic lesions reacted positively with antibodies against Fusobacteriumnecrophorum subsp. necrophorum. Thus, the necrotic lesions were confirmed to be associated with P. aeruginosa and Fusobacterium spp. This is the first detection of P. aeruginosa in bovine esophageal and glossal ulcers associated with Fusobacterium spp.

Childhood vulvovaginitis: report of two cases.

PubMed

Tsai, Hsiu-Yuan; Wan, Chuan; Tseng, Ching-Chang

2006-01-01

Vaginal discharge in young prepubertal girls is a common problem in clinical practice. No specific infective pathogen is identified in most of the children. The reported common microbes include group A beta-hemolytic streptococci, Haemophilus influenzae and some Gram-negative bacilli. Sexually transmitted pathogens such as Neisseria gonorrhoeae and Chlamydia trachomatis are important causes of vulvovaginitis in children suffering from sexual abuse. We report two cases of prepubertal vulvovaginitis presenting with profuse purulent vaginal discharge, and H. influenzae and N. gonorrhoeae identified respectively. Both girls denied any sexual exposure and there was no evidence of sexual abuse. They responded well to antibiotic treatment, and no symptoms recurred in the following months. We would like to emphasize the defined etiology of childhood vulvovaginitis and appropriate treatment, in addition to gynecological evaluation for evidence of sexual abuse.

Pneumonia in the long-term-care facility.

PubMed

Marrie, Thomas J

2002-03-01

Pneumonia is a common infection among residents of long-term-care facilities (LTCFs), with an incidence of 1.2 episodes per 1,000 patient-days. This rate is believed to be six- to tenfold higher than the rate of pneumonia among elderly individuals living in the community. The risk factors for pneumonia among residents of LTCFs are profound disability, bedridden state, urinary incontinence, difficulty swallowing, malnutrition, tube feedings, contractures, and use of benzodiazepines and anticholinergic medications. An elevated respiratory rate is often an early clue to pneumonia in this group of patients. Staphylococcus aureus (including methicillin-resistant S. aureus) and aerobic gram-negative bacilli (including multidrug-resistant isolates) are more frequent causes of pneumonia in this setting than in the community. Criteria have been developed that help identify patients for treatment in their LTCFs.

Anaerobic infections in surgical wards: a two year study

PubMed Central

Ananth-Shenoy, Padmaja; Vishwanath, Shashidhar; Targain, Ryumzook; Shetty, Seema; Sunil-Rodrigues, Gabriel; Mukhopadhyay, Chiranjay; Chawla, Kiran

2016-01-01

Background and Objectives: Anaerobic bacteria are recognized as important pathogens in surgical infections. However, they are the most overlooked microorganisms by the clinic and the laboratory because of the tedious culture techniques with longer turn-around times. The study was aimed to analyze the frequency of anaerobic bacterial surgical infections and their predisposing factors. Materials and Methods: A retrospective study was conducted over a period of two years including patients with surgical infections. The specimens were processed by Gram staining, aerobic and anaerobic culture. The anaerobic bacteria were isolated using standard procedures. The predisposing factors and clinical presentation were studied in these patients. Results: A total of 261 specimens were received from patients with diverse infections from surgical wards. Ninety-one anaerobes were isolated from 64 (24.5%) surgical patients with a predominance of Gram-negative bacilli (37.4%). Anaerobic bacteria as monomicrobial isolates were seen in 21.9% isolates. Anaerobic bacterial isolation along with aerobic bacteria was seen in 71.9% of patients and polymicrobial anaerobic growth was detected in 6.3% of patients. Diabetes mellitus (28, 43.8%) was found to be the most frequent predisposing factor. Bacteroides fragilis group (20.9%) were the most frequent anaerobic Gram-negative bacilli followed by Prevotella spp. (12.1%). Peptostreptococcus anaerobius was the predominant anaerobic cocci isolated (14.3%). Necrotizing fascitis (34.4%) was the most common clinical presentation with anaerobic etiology followed by deep seated abscesses (23.4%). Conclusion: Anaerobic bacteria were isolated from a significant proportion of surgical infections. To avoid therapeutic failures, anaerobic bacteria in surgical infections need to be recognized by surgeons and laboratorians. PMID:27928485

Time-to-positivity, type of culture media and oxidase test performed on positive blood culture vials to predict Pseudomonas aeruginosa in patients with Gram-negative bacilli bacteraemia.

PubMed

Cobos-Triguero, N; Zboromyrska, Y; Morata, L; Alejo, I; De La Calle, C; Vergara, A; Cardozo, C; Arcas, M P; Soriano, A; Marco, F; Mensa, J; Almela, M; Martínez, J A

2017-02-01

The aim of this study was to determine the usefulness of oxidase test and time-to-positivity (TTP) in aerobic and anaerobic blood culture vials to detect the presence of Pseudomonas aeruginosa in patients with Gram-negative bacilli (GNB) bacteraemia. TTP was recorded for each aerobic and anaerobic blood culture vial of monomicrobial bacteraemia due to GNB. Oxidase test was performed in a pellet of the centrifuged content of the positive blood culture. An algorithm was developed in order to perform the oxidase test efficiently taking into account TTP and type of vial. A total of 341 episodes of GNB bacteraemia were analysed. Sensitivity, specificity, positive predictive value and negative predictive value of the oxidase test performed on positive vials with GNB to predict P. aeruginosa were 95%, 99%, 91%, and 99%, respectively. When growth was first or exclusively detected in anaerobic vials, P. aeruginosa was never identified hence the performance of the oxidase test could be avoided. When growth was only or first detected in aerobic vials, a TTP≥8h predicted P. aeruginosa in 37% or cases (63 of 169), therefore oxidase test is highly recommended. Oxidase test performed onto positive blood culture vials previously selected by TTP and type of vials is an easy and inexpensive way to predict P. aeruginosa. In most cases, this can lead to optimization of treatment in less than 24 hours.

Utility of Acridine Orange staining for detection of bacteria from positive blood cultures.

PubMed

Neeraja, M; Lakshmi, V; Padmasri, C; Padmaja, K

2017-08-01

The diagnostic performance of AO stain was evaluated for the detection of bacteria and or fungi from positive blood cultures. The sensitivity of Gram stain (GS) was 98.26% while Acridine Orange (AO) stain proved to be more sensitive (100%) with a Positive and Negative Predictive Value of 100% each. The specificity of both the stains was 100%. Overall agreement between the two stains was 98.23% (688/700). The organisms that were missed by GS and positive by AO were Candida species (Sutton, 2006) and Gram negative bacilli (GNB) (Sutton, 2006). Sensitivity of GS was 82.35% and AO was 100% among mixed cultures. Immediate reporting of the results of AO stain would have a significant impact on clinical management of patients with serious blood stream infections. Copyright © 2017 Elsevier B.V. All rights reserved.

In Vitro Activity of the Siderophore Cephalosporin, Cefiderocol, against a Recent Collection of Clinically Relevant Gram-Negative Bacilli from North America and Europe, Including Carbapenem-Nonsusceptible Isolates (SIDERO-WT-2014 Study).

PubMed

Hackel, Meredith A; Tsuji, Masakatsu; Yamano, Yoshinori; Echols, Roger; Karlowsky, James A; Sahm, Daniel F

2017-09-01

Cefiderocol (formerly S-649266) is an investigational siderophore cephalosporin. Iron-depleted cation-adjusted Mueller-Hinton broth (ID-CAMHB) was prepared according to the Clinical and Laboratory Standards Institute (CLSI) protocol and used to perform broth microdilution testing of cefiderocol against a 2014-2015 collection of clinical isolates of Gram-negative bacilli from North America ( n = 4,239) and Europe ( n = 4,966). The concentrations of cefiderocol inhibiting 90% of isolates tested (MIC 90 s) were 0.5 μg/ml (North America; n = 3,007) and 1 μg/ml (Europe; n = 3,080) for all isolates of Enterobacteriaceae ; 1 μg/ml (North America; n = 30) and 4 μg/ml (Europe; n = 139) for meropenem-nonsusceptible (MIC ≥ 2 μg/ml) isolates of Enterobacteriaceae ; 0.5 μg/ml for both North American ( n = 765) and European ( n = 765) isolates of Pseudomonas aeruginosa ; 0.5 μg/ml (North America; n = 151) and 1 μg/ml (Europe; n = 202) for meropenem-nonsusceptible (MIC ≥ 4 μg/ml) isolates of P. aeruginosa ; 1 μg/ml for both North American ( n = 309) and European ( n = 839) isolates of all Acinetobacter baumannii strains as well as for both North American ( n = 173) and European ( n = 595) isolates of meropenem-nonsusceptible A. baumannii ; and 0.5μg/ml (North America; n = 152) and 0.25 μg/ml (Europe; n = 276) for isolates of Stenotrophomonas maltophilia MICs of cefiderocol were ≤4 μg/ml for 99.9% (6,078/6,087) of all Enterobacteriaceae , 97.0% (164/169) of meropenem-nonsusceptible Enterobacteriaceae , 99.9% (1,529/1,530) of all P. aeruginosa isolates, 100% (353/353) of meropenem-nonsusceptible P. aeruginosa isolates, 97.6% (1,120/1,148) of all A. baumannii isolates, 96.9% (744/768) of meropenem-nonsusceptible A. baumannii isolates, 100% of isolates of S. maltophilia (428/428) and 93.8% of isolates of Burkholderia cepecia (11/12). We conclude that cefiderocol demonstrated potent in vitro activity against a recent collection of clinical isolates of commonly encountered Gram-negative bacilli, including carbapenem-nonsusceptible isolates. Copyright © 2017 American Society for Microbiology.

Occurrence of bla genes encoding carbapenem-resistant Pseudomonas aeruginosa and Acinetobacter baumannii from Intensive Care Unit in a tertiary care hospital.

PubMed

Subramaniyan, Jayanthi Siva; Sundaram, Jeya Meenakshi

2018-01-01

ICU shows increasing incidence of infection associated with the use of invasive procedures for the diagnostic purpose as well as the indiscriminate use of antibiotics. Pseudomonas aeruginosa and Acinetobacter species are "very successful" pathogen and the emergence of the Metallo-β-Lactamases (MBL) is becoming a therapeutic challenge. To isolate the Nonfermenting Gram negative bacilli from the ICU samples. To identify the metallo betalactamase producers and to detect the bla gene presence among the Pseudomonas aeruginosa and Acinetobacter baumannii . The Nonfermenting Gram negative bacilli isolates from the ICU samples were taken over for 5 years (2009-2014) in a tertiary care hospital. The isolates of Pseudomonas species and Acinetobacter species were confirmed by API analyser and processed according to standard procedures. Detection of the MBL producers were done by E strip method and subjected for bla gene detection by PCR method. In our study a total of 195 isolates of NFGNB were obtained from various ICU. Of these MBL producers, 26 % were Pseudomonas aeruginosa and 25 % were Acinetobacter baumannii . The subtypes of bla VIM MBL producing P.aeruginosa were 26%. The predominant gene coding for MBL activity in A.baumannii were found to be bla OXA gene 11.9%. The gene accession numbers were KF975367, KF975372. We have to control the development and dissemination of these superbugs among the ICU's.

The Challenging Diagnosis of Non-Community-Acquired Pneumonia in Non-Mechanically Ventilated Subjects: Value of Microbiological Investigation.

PubMed

Messika, Jonathan; Stoclin, Annabelle; Bouvard, Eric; Fulgencio, Jean-Pierre; Ridel, Christophe; Muresan, Ioan-Paul; Boffa, Jean-Jacques; Bachmeyer, Claude; Denis, Michel; Gounant, Valérie; Esteso, Adoracion; Loi, Valeria; Verdet, Charlotte; Prigent, Hélène; Parrot, Antoine; Fartoukh, Muriel

2016-02-01

Early recognition and an attempt at obtaining microbiological documentation are recommended in patients with non-community-acquired pneumonia (NCAP), whether hospital-acquired (HAP) or health care-associated (HCAP). We aimed to characterize the clinical features and microbial etiologies of NCAP to assess the impact of microbiological investigation on their management. This was a prospective 1-y study in a university hospital with 141 non-mechanically ventilated subjects suspected of having HAP (n = 110) or HCAP (n = 31). Clinical criteria alone poorly identified pneumonia (misdiagnosis in 50% of cases). Microbiological confirmation was achievable in 80 subjects (57%). Among 79 microorganisms isolated, 28 were multidrug-resistant aerobic Gram-negative bacilli and group III Enterobacteriaceae and 6 were methicillin-resistant Staphylococcus aureus. Multidrug-resistant aerobic Gram-negative bacilli accounted for one third of the microorganisms in early-onset HAP and for 50% in late-onset HAP. Methicillin-resistant S. aureus was most often recovered from subjects with HCAP. Inappropriate empirical antibiotics were administered to 36% of subjects with confirmed pneumonia. Forty subjects were admitted to the ICU, 13 (33%) of whom died. Overall, 39 subjects (28%) died in the hospital. Integrating the microbiological investigation in the complex clinical diagnostic workup of patients suspected of having NCAP is mandatory. Respiratory tract specimens should be obtained whenever possible for appropriate management. Copyright © 2016 by Daedalus Enterprises.

Misinterpretation of Gram Stain from the Stationary Growth Phase of Positive Blood Cultures for Brucella and Acinetobacter Species.

PubMed

Bazzi, Ali M; Al-Tawfiq, Jaffar A; Rabaan, Ali A

2017-01-01

Acinetobacter baumannii and Brucella species are Gram-negative organisms that are vulnerable to misinterpretation as Gram-positive or Gram-variable in blood cultures. We assess the random errors in gram stain interpretation to reduce the likelihood of such errors and therefore patient harm. Aerobic and anaerobic blood cultures from two patients in an acute care facility in Saudi Arabia were subjected to preliminary Gram-staining. In case 1, VITEK-2 Anaerobe Identification, repeat Gram staining from a blood agar plate, Remel BactiDrop™ Oxidase test, Urea Agar urease test and real-time PCR were used to confirm presence of Brucella and absence of Coryneform species. In case 2, repeat Gram- staining from the plate and the vials, VITEK-2 Gram-Negative Identification, real-time PCR and subculture on to Columbia agar, blood agar, and MacConkey agar were carried out to identify A. baumannii . In case 1, initially pleomorphic Gram-positive bacteria were identified. Coryneform species were suspected. Tiny growth was observed after 24 h on blood agar plates, and good growth by 48 h. Presence of Brucella species was ultimately confirmed. In case 2, preliminary Gram-stain results suggested giant Gram-positive oval cocci. Further testing over 18-24 h identified A. baumannii . Oxidase test from the plate and urease test from the culture vial is recommended after apparent identification of pleomorphic Gram-positive bacilli from blood culture, once tiny growth is observed, to distinguish Brucella from Corynebacterium species. If giant Gram-positive oval cocci are indicated by preliminary Gram-staining, it is recommended that the Gram stain be repeated from the plate after 4-6 h, or culture should be tested in Triple Sugar Iron (TSI) medium and the Gram stain repeated after 2-4 h incubation.

First case report of sepsis caused by Rhizobium pusense in Japan.

PubMed

Kuchibiro, Tomokazu; Hirayama, Katsuhisa; Houdai, Katsuyuki; Nakamura, Tatsuya; Ohnuma, Kenichirou; Tomida, Junko; Kawamura, Yoshiaki

2018-01-01

Species of the genus Rhizobium are opportunistic, usually saprophytic, glucose-non-fermenting, Gram-negative bacilli found in agricultural soil. Rhizobium pusense infections are the least common Rhizobium infections and have low incidence. Herein, we report the first case of sepsis with R. pusense in Japan in a 67-year-old Japanese woman with a history of hyperlipidaemia, hypertension, diabetes, hypothyroidism and osteoporosis. She had undergone cerebrovascular treatment because she was diagnosed with a subarachnoid haemorrhage. The results of postoperative blood culture showed oxidase-positive, urease-positive, non-lactose-fermenting Gram-stain-negative rods. Using the Vitek2 system, the isolate was distinctly identified as Rhizobium radiobacter . However, 16S rRNA gene sequencing showed 99.93 % similarity with the type strain of R. pusense and 99.06 % similarity with the type strain of R. radiobacter . Additional gene sequencing analysis using recA (97.2 %) and atpD (96.2 %) also showed that the isolated strain is most closely related to R. pusense . The patient was cured by treatment using intravenous meropenem (3 g/d) for 4 weeks and was discharged safely. The definite source of sepsis was unknown. However, the possibility of having been infected through the catheter during the cerebrovascular operation was speculated.

The Assessment of Oral Microflora Exposed to 3% Ethanolic Extract of Brazilian Green Propolis Preparation Used for Hygiene Maintenance following Minor Oral Surgeries

PubMed Central

Morawiec, Tadeusz; Wojtyczka, Robert D.; Niedzielska, Iwona; Bubiłek-Bogacz, Anna; Wróbel, Jacek; Tanasiewicz, Marta; Wesołowski, Piotr; Król, Wojciech

2015-01-01

The aim of this study was to investigate the influence of a topically administered hygienic preparation containing a 3% ethanolic extract of Brazilian green propolis (EEP-B) on oral microflora spectrum changes in a group of patients who underwent common oral surgery procedures. Two gel samples were compared: the tested gel containing an active ingredient, that is, a 3% EEP-B (gel GA), and a placebo as the negative control (gel GC). The collection of microbiological material included 14 patients requiring surgical extraction of wisdom molars and short endosseous implant installation. Clinical examinations were carried out as follow-up, that is, baseline and after 5-6 weeks' time. During the first and subsequent assessment, swabs were taken from the mucosal surface. The number of microorganism species was found to have increased following the application of GC gel over the period of 5-6 weeks. This mainly affected Gram-positive rods and bacilli as well as Gram-negative rods. Application of the GA gel enriched with 3% EEP-B caused a profound reduction in the amount of Neisseria spp. and Bifidobacterium spp. strains. Elimination of seven species of microorganisms was observed: Streptococcus acidominimus, Streptococcus oralis, Staphylococcus epidermidis, Veillonella parvula, Bifidobacterium breve, Bifidobacterium longum, and Lactobacillus acidophilus. PMID:26380300

Antibacterial metabolites synthesized by psychrotrophic bacteria isolated from cold-freshwater environments.

PubMed

Barros, Javier; Becerra, José; González, Carlos; Martínez, Miguel

2013-03-01

The ability of three psychrotrophic Gram-negative bacilli isolated from Chilean Patagonian cold freshwater rivers to produce bioactive metabolites was evaluated. The strains were isolated from cold waters rivers and identified by their biochemical properties and 16S rRNA gene analysis. The metabolites fractions showing antibacterial activity were obtained by solvent extraction and partially characterized by gas-mass chromatography (GC-MS). Antibacterial activity of the fractions was evaluated by an agar-well diffusion test upon 14 bacterial strains, both Gram positive and Gram negative. Thermal and proteolytic resistances of the antibacterial metabolites fractions were also evaluated. Molecular analysis allows the identification of the three Patagonian strains as Pseudomonas sp. RG-6 (Pseudomonas brenneri 99.6 % identity), Pseudomonas sp. RG-8 (Pseudomonas trivialis 99.6 % identity) and Yersinia sp. RP-3 (Yersinia aldovae 99.5 % identity). These extracts were able to inhibit both Gram-positive and Gram-negative bacteria but not Listeria monocytogenes. The antibacterial activity of the filtrated supernatants was lost at temperatures ≥60 °C, and was not affected by proteinase K treatment. The chemical structure of the active molecule remains to be elucidated, although the GC-MS analysis of the filtrates suggests that compounds like sesquiterpenes derivatives from β-maaliene or δ-selinene could be responsible of this antibacterial activity. Pristine cold freshwater streams showed to be interesting sources of metabolites-producing microorganisms with antibacterial activity.

Surveillance of antimicrobial susceptibility of aerobic and facultative Gram-negative bacilli isolated from patients with intra-abdominal infections in China: the 2002-2009 Study for Monitoring Antimicrobial Resistance Trends (SMART).

PubMed

Yang, Qiwen; Wang, Hui; Chen, Minjun; Ni, Yuxing; Yu, Yunsong; Hu, Bijie; Sun, Ziyong; Huang, Wenxiang; Hu, Yunjian; Ye, Huifen; Badal, Robert E; Xu, Yingchun

2010-12-01

The objective of this study was to investigate the distribution and susceptibility of aerobic and facultative Gram-negative bacilli (GNB) isolated from patients with intra-abdominal infections (IAIs) in China. From 2002 to 2009, minimum inhibitory concentrations of 14 antibiotics for 3420 aerobic and facultative GNB from up to eight hospitals in six cities were determined by the broth microdilution method. Enterobacteriaceae comprised 82.9% (2834/3420) of the total isolates, with Escherichia coli (49.2%) being the most commonly isolated species followed by Klebsiella pneumoniae (17.0%), Enterobacter cloacae (5.8%) and Citrobacter freundii (2.3%). Amongst the antimicrobial agents tested, the three carbapenems (ertapenem, imipenem and meropenem) were the most active agents against Enterobacteriaceae, with susceptibility rates of 96.1-99.6% (2002-2009), 98.2-100% (2002-2009) and 99.6-100% (2002-2004), respectively, followed by amikacin (86.8-95.1%) and piperacillin/tazobactam (84.5-94.3%). Susceptibility rates of all tested third- and fourth-generation cephalosporins against Enterobacteriaceae declined by nearly 30%, with susceptibility rates of 40.2%, 39.1%, 56.3% and 51.8% in 2009 for ceftriaxone, cefotaxime, ceftazidime and cefepime, respectively. The occurrence of extended-spectrum β-lactamases increased rapidly, especially for E. coli (from 20.8% in 2002 to 64.9% in 2009). Susceptibility of E. coli to ciprofloxacin decreased from 57.6% in 2002 to 24.2% in 2009. The least active agent against Enterobacteriaceae was ampicillin/sulbactam (SAM) (25.3-44.3%). In conclusion, Enterobacteriaceae were the major pathogens causing IAIs, and carbapenems retained the highest susceptibility rates over the 8-year study period. Third- and fourth-generation cephalosporins, fluoroquinolones and SAM may not be ideal choices for empirical therapy of IAIs in China. Copyright © 2010 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

Assessment of airborne bacteria in selected occupational environments in Quezon City, Philippines.

PubMed

Rendon, Rhoshela Vi C; Garcia, Bea Clarise B; Vital, Pierangeli G

2017-05-04

Exposure to bioaerosols has been associated with health deterioration among workers in several occupational environments. This highlights the need to study the microbiological quality of air of workplaces as no such study has been conducted yet in the Philippines. To detect and characterize the culturable mesophilic airborne bacteria in selected occupational environments we used passive sedimentation technique. It was observed that the number of colony-forming units was highest in junk shop, followed by the light railway transit station and last the office. By contrast, the bacterial composition was similar in all sites: Gram-positive cocci > Gram-positive bacilli > Gram-negative bacteria. Staphylococcus aureus and Bacillus spp. were also detected in all sites. These findings suggest that the presence of airborne bacteria may be a potential health hazard in urban occupational environments in the Philippines.

Screening of nursing home residents for colonization with carbapenem-resistant Enterobacteriaceae admitted to acute care hospitals: Incidence and risk factors.

PubMed

Cunha, Cheston B; Kassakian, Steven Z; Chan, Ryan; Tenover, Fred C; Ziakas, Panos; Chapin, Kimberle C; Mermel, Leonard A

2016-02-01

There are increasing reports of multidrug-resistant gram-negative bacilli in nursing homes and acute care hospitals. We performed a point prevalence survey to detect fecal carriage of gram-negative bacteria carrying carbapenem resistance genes or which were otherwise resistant to carbapenem antibiotics among 500 consecutive admissions from local nursing homes to 2 hospitals in Providence, Rhode Island. We performed a case-control study to identify risk factors associated with carriage of carbapenem-resistant Enterobacteriaceae (CRE). There were 404 patients with 500 hospital admissions during which they had rectal swab samples cultured. Fecal carriage of any carbapenem-resistant or carbapenemase- producing gram-negative bacteria was found in 23 (4.6%) of the 500 hospital admissions, including 7 CRE (1.4%), 2 (0.4%) of which were Klebsiella pneumoniae carbapenemase (ie, blaKPC) producing (CPE) Citrobacter freundii, 1 of which was carbapenem susceptible by standard testing methods. Use of a gastrostomy tube was associated with CRE carriage (P = .04). We demonstrated fecal carriage of carbapenem-resistant or carbapenemase-producing gram-negative bacteria in 4.6% of nursing home patients admitted to 2 acute care hospitals, but only 0.4% of such admissions were patients with fecal carriage of CPE. Use of gastrostomy tubes was associated with fecal carriage of gram-negative bacteria with detectable carbapenem resistance. CRE fecal carriage is uncommon in our hospital admissions from nursing homes. Copyright © 2016 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

Antimicrobial susceptibility of clinically isolated anaerobic bacteria in a University Hospital Centre Split, Croatia in 2013.

PubMed

Novak, Anita; Rubic, Zana; Dogas, Varja; Goic-Barisic, Ivana; Radic, Marina; Tonkic, Marija

2015-02-01

Anaerobic bacteria play a significant role in many endogenous polymicrobial infections. Since antimicrobial resistance among anaerobes has increased worldwide, it is useful to provide local susceptibility data to guide empirical therapy. The present study reports recent data on the susceptibility of clinically relevant anaerobes in a University Hospital Centre (UHC) Split, Croatia. A total of 63 Gram-negative and 59 Gram-positive anaerobic clinical isolates from various body sites were consecutively collected from January to December 2013. Antimicrobial susceptibility testing was performed using standardized methods and interpreted using EUCAST criteria. Patient's clinical and demographic data were recorded by clinical microbiologist. Among 35 isolates of Bacteroides spp., 97.1% were resistant to penicillin (PCN), 5.7% to amoxicillin/clavulanic acid (AMC), 8.6% to piperacillin/tazobactam (TZP), 29.0% to clindamycin (CLI) and 2.9% to metronidazole (MZ). Percentages of susceptible strains to imipenem (IPM), meropenem (MEM) and ertapenem (ETP) were 94.3. Resistance of other Gram-negative bacilli was 76.0% to PCN, 8.0% to AMC, 12.0% to TZP, 28.0% to CLI and 8% to MZ. All other Gram-negative strains were fully susceptible to MEM and ETP, while 96.0% were susceptible to IPM. Clostridium spp. isolates were 100% susceptible to all tested antibiotics except to CLI (two of four tested isolates were resistant). Propionibacterium spp. showed resistance to CLI in 4.3%, while 100% were resistant to MZ. Among other Gram-positive bacilli, 18.2% were resistant to PCN, 9.1% to CLI and 54.5% to MZ, while 81.8% of isolates were susceptible to carbapenems. Gram-positive cocci were 100% susceptible to all tested antimicrobials except to MZ, where 28.6% of resistant strains were recorded. Abdomen was the most common source of isolates (82.5%). The most prevalent types of infection were abscess (22.1%), sepsis (14.8%), appendicitis (13.9%) and peritonitis (6.6%). Twenty four patients (19.7%) received empiric antimicrobial therapy. One hundred and one patients (82.8%) had polymicrobial aerobic/anaerobic isolates cultivated from the same specimens. Almost all aerobic bacteria were of endogenous origin and showed fully susceptible antimicrobial profile; only 8.7% (9/104) were multiresistant and considered as hospital acquired. Based on our findings, β-lactam/β-lactamase inhibitor combinations and metronidazole remain useful antimicrobials for empiric treatment of anaerobic infections, while carbapenems should be reserved for situations were multidrug resistant, aerobic or facultative Gram-negative bacteria are expected. However, a certain percentage of resistant isolates were observed for each of these agents. Therefore, periodic resistance surveillance in anaerobes is highly recommended in order to guide empirical therapy. Copyright © 2014 Elsevier Ltd. All rights reserved.

Identification and Susceptibility Testing of Enterobacteriaceae and Pseudomonas aeruginosa by Direct Inoculation from Positive BACTEC Blood Culture Bottles into Vitek 2

PubMed Central

Bruins, Marjan J.; Bloembergen, Peter; Ruijs, Gijs J. H. M.; Wolfhagen, Maurice J. H. M.

2004-01-01

Inoculation of an automated system for rapid identification (ID) and antimicrobial susceptibility testing (AST) directly from positive blood culture bottles will reduce the turnaround time of laboratory diagnosis of septicemic patients, which benefits clinical outcome and decreases patient costs. Direct test results, however, must always be confirmed by testing a pure overnight culture, which is the “gold standard.” We studied the accuracy of direct testing versus repeat testing in order to investigate the possibility of refraining from repeat testing. We also assessed the clinical risk of reporting results based on direct testing only. We inoculated Vitek 2 (bioMérieux) directly from 410 positive BACTEC 9240 (BD) blood culture bottles containing gram-negative rods and studied the ID and AST results. In a comparison of direct inoculation with the standard method, a total of 344 isolates of Enterobacteriaceae and Pseudomonas aeruginosa were tested, and 93.0% were correctly identified. Of the 39 (10.2%) samples that contained bacilli not identifiable by Vitek 2, only 1 gave a conclusive, correct result. The overall MIC agreement among 312 isolates was 99.2%, with 0.8% very major and 0.02% major error rates. Of only three (polymicrobial) samples, the direct susceptibility pattern would be reported to the clinician as too sensitive. Vitek 2 results obtained from direct inoculation of blood culture bottles containing gram-negative bacilli are safe enough for immediate reporting, provided that ID and AST are consistent. Repeat testing is not necessary, unless Gram stain or overnight subculture results raise doubt about the purity of the culture. PMID:14715724

Late-onset neonatal sepsis: recent developments

PubMed Central

Dong, Ying; Speer, Christian P

2015-01-01

The incidence of neonatal late-onset sepsis (LOS) is inversely related to the degree of maturity and varies geographically from 0.61% to 14.2% among hospitalised newborns. Epidemiological data on very low birth weight infants shows that the predominant pathogens of neonatal LOS are coagulase-negative staphylococci, followed by Gram-negative bacilli and fungi. Due to the difficulties in a prompt diagnosis of LOS and LOS-associated high risk of mortality and long-term neurodevelopmental sequelae, empirical antibiotic treatment is initiated on suspicion of LOS. However, empirical therapy is often inappropriately used with unnecessary broad-spectrum antibiotics and a prolonged duration of treatment. The increasing number of multidrug-resistant Gram-negative micro-organisms in neonatal intensive care units (NICU) worldwide is a serious concern, which requires thorough and efficient surveillance strategies and appropriate treatment regimens. Immunological strategies for preventing neonatal LOS are not supported by current evidence, and approaches, such as a strict hygiene protocol and the minimisation of invasive procedures in NICUs represent the cornerstone to reduce the burden of neonatal LOS. PMID:25425653

In vitro activity of imipenem/relebactam against Gram-negative ESKAPE pathogens isolated in 17 European countries: 2015 SMART surveillance programme.

PubMed

Karlowsky, James A; Lob, Sibylle H; Kazmierczak, Krystyna M; Hawser, Stephen P; Magnet, Sophie; Young, Katherine; Motyl, Mary R; Sahm, Daniel F

2018-04-11

Relebactam is an inhibitor of class A β-lactamases, including KPC β-lactamases, and class C β-lactamases, and is currently under clinical development in combination with imipenem. The objective of the current study was to evaluate the in vitro activity of imipenem/relebactam against Gram-negative ESKAPE pathogens (Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.) submitted by clinical laboratories in 17 European countries to the Study for Monitoring Antimicrobial Resistance Trends (SMART) global surveillance programme in 2015. MICs were determined using the CLSI standard broth microdilution method and interpreted using EUCAST clinical breakpoints. Relebactam was tested at a fixed concentration of 4 mg/L in combination with doubling dilutions of imipenem. Imipenem/relebactam MICs were interpreted using breakpoints for imipenem. Rates of susceptibility to imipenem and imipenem/relebactam for isolates of P. aeruginosa (n = 1705), K. pneumoniae (n = 1591) and Enterobacter spp. (n = 772) were 72.0/94.7%, 88.7/94.8% and 95.6/96.8%, respectively. Relebactam restored imipenem susceptibility to 81.1%, 54.2% and 26.5% of imipenem-non-susceptible isolates of P. aeruginosa (n = 477), K. pneumoniae (n = 179) and Enterobacter spp. (n = 34). Most imipenem/relebactam-non-susceptible isolates carried MBLs, OXA-48 or GES carbapenemases. Relebactam did not increase the number of isolates of A. baumannii (n = 486) susceptible to imipenem. Relebactam restored susceptibility to imipenem for the majority of imipenem-non-susceptible isolates of P. aeruginosa and K. pneumoniae tested as well as some isolates of imipenem-non-susceptible Enterobacter spp. Based on our results, imipenem/relebactam appears to be a promising therapeutic option for treating patients with infections caused by antimicrobial-resistant Gram-negative bacilli.

The Genus Corynebacterium and Other Medically Relevant Coryneform-Like Bacteria

PubMed Central

2012-01-01

Catalase-positive Gram-positive bacilli, commonly called “diphtheroids” or “coryneform” bacteria were historically nearly always dismissed as contaminants when recovered from patients, but increasingly have been implicated as the cause of significant infections. These taxa have been underreported, and the taxa were taxonomically confusing. The mechanisms of pathogenesis, especially for newly described taxa, were rarely studied. Antibiotic susceptibility data were relatively scant. In this minireview, clinical relevance, phenotypic and genetic identification methods, matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) evaluations, and antimicrobial susceptibility testing involving species in the genus Corynebacterium and other medically relevant Gram-positive rods, collectively called coryneforms, are described. PMID:22837327

Isolation and whole genome analysis of endospore-forming bacteria from heroin.

PubMed

Kalinowski, Jörn; Ahrens, Björn; Al-Dilaimi, Arwa; Winkler, Anika; Wibberg, Daniel; Schleenbecker, Uwe; Rückert, Christian; Wölfel, Roman; Grass, Gregor

2018-01-01

Infections caused by endospore-forming bacteria have been associated with severe illness and death among persons who inject drugs. Analysis of the bacteria residing in heroin has thus been biased towards species that affect human health. Similarly, exploration of the bacterial diversity of seized street market heroin correlated with the skin microflora of recreational heroin users insofar as different Staphylococus spp. or typical environmental endospore formers including Bacillus cereus and other Bacilli outside the B. cereus sensu lato group as well as diverse Clostridia were identified. In this work 82 samples of non-street market ("wholesale") heroin originating from the German Federal Criminal Police Office's heroin analysis program seized during the period between 2009 and 2014 were analyzed for contaminating bacteria. Without contact with the end user and with only little contaminations introduced by final processing, adulteration and cutting this heroin likely harbors original microbiota from the drug's original source or trafficking route. We found this drug to be only sparsely populated with retrievable heterotrophic, aerobic bacteria. In total, 68 isolates were retrieved from 49 out of 82 samples analyzed (60% culture positive). All isolates were endospore-forming, Gram-positive Bacilli. Completely absent were non-endospore-formers or Gram-negatives. The three most predominant species were Bacillus clausii, Bacillus (para)licheniformis, and Terribacillus saccharophilus. Whole genome sequencing of these 68 isolates was performed using Illumina technology. Sequence data sets were assembled and annotated using an automated bioinformatics pipeline. Average nucleotide identity (ANI) values were calculated for all draft genomes and all close to identical genomes (ANI>99.5%) were compared to the forensic data of the seized drug, showing positive correlations that strongly warrant further research on this subject. Copyright © 2017 Elsevier B.V. All rights reserved.

THE OCCURRENCE OF POLYGLYCEROPHOSPHATE AS AN ANTIGENIC COMPONENT OF VARIOUS GRAM-POSITIVE BACTERIAL SPECIES

PubMed Central

McCarty, Maclyn

1959-01-01

A bacterial substance has been described which gives a precipitin reaction with certain antisera to Group A streptococci. The precipitating antigen is present in various Gram-positive bacteria, including most hemolytic streptococci, staphylococci, and aerobic sporulating bacilli. It is not present in any of the Gram-negative species examined or in pneumococci, clostridia, or corynebacteria. Analysis of purified preparations obtained from Group A streptococci indicates that the antigen is a simple polymer of glycerophosphate. The identification has been confirmed by immunochemical studies, including precipitin tests and specific inhibition with synthetic polyglycerophosphates. In addition, the infrared spectra of bacterial and synthetic polyglycerophosphate are shown to be closely similar. Immunochemical analysis suggests that the amount of polyglycerophosphate present in Group A streptococci and staphylococci is approximately 1 per cent of the dry weight of the cells. The cellular localization and function of the polyglycerophosphate have not been established. PMID:13641562

Identification of Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa in Blood Cultures: a Multicenter Performance Evaluation of a Three-Color Peptide Nucleic Acid Fluorescence In Situ Hybridization Assay▿

PubMed Central

Della-Latta, Phyllis; Salimnia, Hossein; Painter, Theresa; Wu, Fann; Procop, Gary W.; Wilson, Deborah A.; Gillespie, Wendy; Mender, Alayna; Crystal, Benjamin S.

2011-01-01

A multicenter evaluation was undertaken to evaluate the performance of a new three-color peptide nucleic acid fluorescence in situ hybridization assay that identifies isolates directly from blood cultures positive for Gram-negative bacilli (GNB). The assay correctly identified 100% (186/186) of the Escherichia coli isolates, 99.1% (109/110) of the Klebsiella pneumoniae isolates, and 95.8% (46/48) of the Pseudomonas aeruginosa isolates in this study. Negative assay results were correctly obtained for 162 of 165 other GNB (specificity, 98.2%). PMID:21490185

Pathogen occurrence and antimicrobial resistance trends among urinary tract infection isolates in the Asia-Western Pacific Region: report from the SENTRY Antimicrobial Surveillance Program, 1998-1999.

PubMed

Turnidge, John; Bell, Jan; Biedenbach, Douglas J; Jones, Ronald N

2002-07-01

Worldwide surveillance of antimicrobial resistance among urinary tract pathogens is useful to determine important trends and geographical variation for common Gram-positive and -negative species. The most common causative uropathogens often have intrinsic or acquired resistance mechanisms which include ESBL production among enteric bacilli, multi-drug resistant staphylococci and non-fermentative Gram-negative bacilli such as Pseudomonas aeruginosa and Acinetobacter spp. and vancomycin-resistant Enterococcus spp. This study evaluates pathogen frequency and the resistance rates among urinary tract infection (UTI) pathogens in 14 medical centres in the Asia-Pacific region between 1998 and 1999. The isolates were referred to a central monitor for reference NCCLS broth microdilution testing, identification confirmation and patient demographic analysis. Over 50% of the 958 pathogens were Escherichia coli and Klebsiella spp. followed by P. aeruginosa, Enterococcus spp. and Enterobacter spp. Susceptibility for the three enteric bacilli was high for carbapenems (100%), 'fourth-generation' cephalosporins (cefepime 94.9-98.6%) and amikacin (> or = 93.0%). Beta-lactamase inhibitor compounds were more active against E. coli (piperacillin/tazobactam; > 90% susceptible) than the other two enteric species and all other tested agents had a narrower spectra of activity. The rank order of anti-pseudomonal agents was amikacin (91.5% susceptible)> imipenem > piperacillin/tazobactam > tobramycin > ceftazidime and cefepime (77.4 and 76.4% susceptible, respectively). Susceptibility to quinolones for the P. aeruginosa isolates was only 63.2-67.0%. Only one vancomycin-intermediate Enterococcus spp. (van C phenotype) was detected among the 103 strains tested. Newer fluoroquinolones (gatifloxacin; MIC(50), mg/l) were more potent against enterococci than ciprofloxacin (MIC(50), 2 mg/l) and high-level resistance to aminoglycosides was common (41.7%). The data presented are compared to studies of similar design from other areas which are part of the SENTRY surveillance network.

Monomicrobial necrotizing fasciitis in a single center: the emergence of Gram-negative bacteria as a common pathogen.

PubMed

Yahav, D; Duskin-Bitan, H; Eliakim-Raz, N; Ben-Zvi, H; Shaked, H; Goldberg, E; Bishara, J

2014-11-01

Necrotizing fasciitis (NF) is a life-threatening soft tissue infection. It is usually caused by Streptococcus pyogenes and other Gram-positive bacteria. Several reports, however, emphasize the importance of Gram-negative rods in this infection. We retrospectively studied all cases of monomicrobial necrotizing fasciitis hospitalized in our center during the years 2002-2012. We compared clinical characteristics and outcomes of patients with Gram-negative versus Gram-positive infection. Forty-five cases were reviewed, 19 caused by Gram-negative organisms, 10 of them Escherichia coli, and 26 caused by Gram-positive organisms, 10 of them S. pyogenes. Compared to Gram-positive infections, patients with Gram-negative infections were more likely to have a baseline malignancy (9/19, 47.4%) or to have undergone recent surgery (4/19, 42.3%). The 30-day mortality was higher among Gram-negative infected patients (8/19, 42.1% vs. 8/26, 30.8%). Creatine phosphokinase (CPK) was elevated in a minority of patients with Gram-negative necrotizing fasciitis, and its absolute value was lower than in Gram-positive necrotizing fasciitis. In our center, 42% of monomicrobial necrotizing fasciitis cases were found to be caused by Gram-negative organisms, mostly E. coli. These infections usually appeared in immunocompromised or postoperative patients, often presented with normal CPK levels, and were associated with high mortality rates.

Direct Antimicrobial Susceptibility Testing of Gram-Negative Bacilli in Blood Cultures by an Electrochemical Method

PubMed Central

Huang, Ay Huey; Wu, Jiunn Jong; Weng, Yu Mei; Ding, Hwia Cheng; Chang, Tsung Chain

1998-01-01

Nonfastidious aerobic gram-negative bacilli (GNB) are commonly isolated from blood cultures. The feasibility of using an electrochemical method for direct antimicrobial susceptibility testing of GNB in positive blood cultures was evaluated. An aliquot (10 μl) of 1:10-diluted positive blood cultures containing GNB was inoculated into the Bactometer module well (bioMérieux Vitek, Hazelwood, Mo.) containing 1 ml of Mueller-Hinton broth supplemented with an antibiotic. Susceptibility tests were performed in a breakpoint broth dilution format, with the results being categorized as resistant, intermediate, or susceptible. Seven antibiotics (ampicillin, cephalothin, gentamicin, amikacin, cefamandole, cefotaxime, and ciprofloxacin) were used in this study, with each agent being tested at the two interpretive breakpoint concentrations. The inoculated modules were incubated at 35°C, and the change in impedance in each well was continuously monitored for 24 h by the Bactometer. The MICs of the seven antibiotics for each blood isolate were also determined by the standardized broth microdilution method. Of 146 positive blood cultures (1,022 microorganism-antibiotic combinations) containing GNB tested by the direct method, the rates of very major, major, and minor errors were 0, 1.1, and 2.5%, respectively. The impedance method was simple; no centrifugation, preincubation, or standardization of the inocula was required, and the susceptibility results were normally available within 3 to 6 h after inoculation. The rapid method may allow proper antimicrobial treatment almost 30 to 40 h before the results of the standard methods are available. PMID:9738038

Occurrence of bla genes encoding carbapenem-resistant Pseudomonas aeruginosa and Acinetobacter baumannii from Intensive Care Unit in a tertiary care hospital

PubMed Central

Subramaniyan, Jayanthi Siva; Sundaram, Jeya Meenakshi

2018-01-01

CONTEXT: ICU shows increasing incidence of infection associated with the use of invasive procedures for the diagnostic purpose as well as the indiscriminate use of antibiotics. Pseudomonas aeruginosa and Acinetobacter species are “very successful” pathogen and the emergence of the Metallo-β-Lactamases (MBL) is becoming a therapeutic challenge. AIMS: To isolate the Nonfermenting Gram negative bacilli from the ICU samples. To identify the metallo betalactamase producers and to detect the bla gene presence among the Pseudomonas aeruginosa and Acinetobacter baumannii. SETTINGS AND DESIGN: The Nonfermenting Gram negative bacilli isolates from the ICU samples were taken over for 5 years (2009-2014) in a tertiary care hospital. METHODS AND MATERIALS: The isolates of Pseudomonas species and Acinetobacter species were confirmed by API analyser and processed according to standard procedures. Detection of the MBL producers were done by E strip method and subjected for bla gene detection by PCR method. RESULTS: In our study a total of 195 isolates of NFGNB were obtained from various ICU. Of these MBL producers, 26 % were Pseudomonas aeruginosa and 25 % were Acinetobacter baumannii. The subtypes of blaVIM MBL producing P.aeruginosa were 26%. The predominant gene coding for MBL activity in A.baumannii were found to be blaOXA gene 11.9%. The gene accession numbers were KF975367, KF975372. CONCLUSIONS: We have to control the development and dissemination of these superbugs among the ICU's. PMID:29692589

Evolving Understanding of the Causes of Pneumonia in Adults, With Special Attention to the Role of Pneumococcus.

PubMed

Musher, Daniel M; Abers, Michael S; Bartlett, John G

2017-10-30

Before 1945, Streptococcus pneumoniae caused more than 90% of cases of pneumonia in adults. After 1950, the proportion of pneumonia caused by pneumococcus began to decline. Pneumococcus has continued to decline; at present, this organism is identified in fewer than fewer10%-15% of cases. This proportion is higher in Europe, a finding likely related to differences in vaccination practices and smoking. Gram-negative bacilli, Staphylococcus aureus, Chlamydia, Mycoplasma, and Legionella are each identified in 2%-5% of patients with pneumonia who require hospitalization. Viruses are found in 25% of patients, up to one-third of these have bacterial coinfection. Recent studies fail to identify a causative organism in more than 50% of cases, which remains the most important challenge to understanding lower respiratory infection. Our findings have important implications for antibiotic stewardship and should be considered as new policies for empiric pneumonia management are developed. © The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.

Evaluation of the β-CARBA™ test, a colorimetric test for the rapid detection of carbapenemase activity in Gram-negative bacilli.

PubMed

Bernabeu, Sandrine; Dortet, Laurent; Naas, Thierry

2017-06-01

There is an urgent need for accurate and fast diagnostic tests to identify carbapenemase-producing bacteria. Here, we have evaluated a novel colorimetric test (the β-CARBA™ test; Bio-Rad) to detect carbapenemase-producing Gram-negative bacilli from cultured colonies. The performance of the β-CARBA™ test was compared with that of the Carba NP test (or the CarbAcineto NP test) and RAPIDEC ® CARBA NP (bioMérieux) using a collection of 290 isolates with characterized β-lactamase content. This collection included 199 carbapenemase producers (121 Enterobacteriaceae, 36 Pseudomonas and 42 Acinetobacter baumannii ) and 91 non-carbapenemase producers (55 Enterobacteriaceae, 20 Pseudomonas and 16 A. baumannii ). The β-CARBA™ test correctly detected 84.9% of the carbapenemase producers, including all KPC and IMP, 96.4% of VIM, 85.3% of NDM, 80.5% of OXA-48-like and 91.2% of A. baumannii -related OXA carbapenemases (OXA-23, OXA-40, OXA-58, OXA-143 and overexpressed OXA-51). All rare metallo-β-lactamases (SPM, AIM, GIM, DIM and SIM) were detected. Importantly, all non-KPC Ambler class A carbapenemases were not detected, including GES variants with carbapenemase activity ( n  = 6), IMI ( n  = 3), NMC-A ( n  = 1), SME ( n  = 2), FRI-1 ( n  = 1) and BIC-1 ( n  = 1). All non-carbapenemase producers gave a negative result except with OXA-163-, OXA-405- and one TEM-3-producing Citrobacter freundii . The overall sensitivity and specificity of the β-CARBA™ test were 84.9% and 95.6%, respectively. This test is easy to perform and to interpret by non-specialized staff members. Despite lack of specificity towards non-KPC Ambler class A and OXA-48-like carbapenemases, the β-CARBA™ test could complete the existing panel of tests available for the confirmation of carbapenemases in Gram-negatives. © The Author 2017. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Polymicrobial nature of vaginitis in young women: a microbiological and therapeutic study.

PubMed

Kippax, R A; Caradoc-Davies, G; Meech, R J

1982-03-24

Thirty-six young females attending the Student Health Service with vaginitis were investigated by serial semiquantitative aerobic, anaerobic, fungal, mycoplasma and viral cultures over a 10 day period and results were correlated with signs and symptoms. Antifungal therapy (econazole pessaries and cream) resulted in clearance of candida from 13 out of 16 patients where there was no increase in the anaerobic flora. In the four subjects where candida was isolated along with Gardnerella vaginalis plus abnormal anaerobic flora, only one cleared with econazole, the remaining three clearing during therapy with metronidazole. In the nine subjects with Gardnerella vaginalis and abnormal anaerobic flora, metronidazole relieved symptoms despite failure to eradicate G. vaginalis in seven indicating the pathogenic role of the anaerobic flora rather then G. vaginalis. Mycoplasma hominis, Ureaplasma urealyticum and gram negative enteric bacilli were not implicated as primary agents in causing vaginitis.

Antimicrobial-resistant Klebsiella species isolated from free-range chicken samples in an informal settlement.

PubMed

Fielding, Burtram C; Mnabisa, Amanda; Gouws, Pieter A; Morris, Thureyah

2012-02-29

Sub-therapeutic doses of antimicrobial agents are administered routinely to poultry to aid growth and to prevent disease, with prolonged exposure often resulting in bacterial resistance. Crossover of antibiotic resistant bacteria from poultry to humans poses a risk to human health. In this study, 17 chicken samples collected from a vendor operating in an informal settlement in the Cape Town Metropolitan area, South Africa were screened for antimicrobial-resistant Gram-negative bacilli using the Kirby Bauer disk diffusion assay. IN TOTAL, SIX ANTIBIOTICS WERE SCREENED: ampicillin, ciprofloxacin, gentamicin, nalidixic acid, tetracycline and trimethoprim. Surprisingly, Klebsiella ozaenae was identified in 96 and K. rhinoscleromatis in 6 (n=102) of the samples tested. Interestingly, ∼40% of the isolated Klebsiella spp. showed multiple resistance to at least three of the six antibiotics tested. Klebsiella ozaenae and K. rhinoscleromatis cause clinical chronic rhinitis and are almost exclusively associated with people living in areas of poor hygiene.

Bloodstream infections in patients with solid tumors

PubMed Central

Gudiol, Carlota; Aguado, José María; Carratalà, Jordi

2016-01-01

ABSTRACT Little information is currently available regarding bloodstream infection (BSI) in patients with solid tumors who, for a variety of reasons, are particularly predisposed to develop this condition. In this review we focus on the incidence, epidemiology, clinical features, etiology, antimicrobial resistance, and outcomes of BSI of adult cancer patients with solid tumors. Most episodes of BSI occur in non-neutropenic patients, in whom the site of primary or metastatic tumor often serves as the portal of entry. The urinary tract and the abdomen are the most frequent sources of infection, and cholangitis is the most common recurrent source of BSI. Gram-negative bacilli are becoming the leading cause of BSI in patients with solid tumors, and the rate of multidrug resistance is increasingly being recognized. The case-fatality rate in patients with solid tumors and BSI is high, especially among those with comorbidities, advanced neoplasms, corticosteroid therapy, and shock at presentation. PMID:26787095

Relative Frequency, Characteristics, and Antimicrobial Susceptibility Patterns of Vibrio spp., Aeromonas spp., Chromobacterium violaceum, and Shewanella spp. in the Northern Territory of Australia, 2000–2013

PubMed Central

McAuliffe, Gary N.; Hennessy, Jann; Baird, Robert W.

2015-01-01

Vibrio, Aeromonas, Chromobacterium violaceum, and Shewanella (VACS) are water-associated Gram-negative organisms that can cause a variety of infections. The frequency, patient characteristics, and antimicrobial susceptibilities for 468 isolates from 442 patients from the Northern Territory were reviewed. Aeromonas spp. (312 of 468; 67%) were most commonly isolated followed by Vibrio spp. (71 of 468; 15%), Shewanella spp. (61 of 468; 13%), and C. violaceum (24 of 468; 5%). A strong male predominance was found (male to female ratio of 2.3:1). Skin and soft tissue isolations (373 of 468; 80%) from lower limb infections (222 of 371; 60%) were the most common clinical manifestation. The episodes were usually polymicrobial (281 of 468; 60%). Coisolates included Staphylococcus aureus (137 of 468; 29%), β-hemolytic streptococci (74 of 468; 16%), enterobacteriaceae (111 of 468; 24%), non-fermentative Gram-negative bacilli (35 of 468; 7%), and other VACS organisms (37 of 468; 8%). Antimicrobial resistance of VACS organisms to ciprofloxacin (0–4%), cefepime (0–3%), and gentamicin (0–0.8%) and Vibrio spp., Aeromonas spp., and Shewanella to cotrimoxazole (0–3%) was rarely shown. For water-associated lower limb skin and soft tissue infections in the tropics, clinicians should consider empirical antimicrobial therapy with agents active against S. aureus and VACS organisms. PMID:25548380

Antimicrobial Activity of a Novel Vascular Access Film Dressing Containing Chlorhexidine Gluconate.

PubMed

Wibaux, Anne; Thota, Priyaleela; Mastej, Jozef; Prince, Daniel L; Carty, Neal; Johnson, Peter

2015-01-01

Covering insertion sites with chlorhexidine impregnated dressings has been proven to be clinically effective in reducing catheter related blood stream infections (CR-BSI). Two chlorhexidine gluconate (CHG)-impregnated dressings are commercially available, a polyurethane foam disk and a film dressing containing a chlorhexidine gluconate-impregnated gel pad. While both have demonstrated efficacy in clinical settings, the major drawback of high cost and impaired IV insertion site visibility limits their usage. A new, simple film dressing containing CHG within its adhesive layer is now available. The objective of this study was to test the in vitro antimicrobial efficacy of the new dressing in comparison to the CHG-impregnated gel dressing. Quantitative aliquots of suspensions (concentration of 1.0x106 to 5.0x106 cfu/sample) of clinically relevant challenge organisms (Staphylococcus species, gram-negative bacilli, Candida albicans) were incubated in contact with the new CHG-containing film dressing, a placebo version of the same (negative control) and the commercially available CHG-impregnated gel dressing (positive control). Serial dilutions of the surviving organisms were quantified using the pour plate after 1, 3, 5, and 7 days of incubation in order to calculate an antimicrobial log10 reduction for each organism/dressing combination at each point in time. The new CHG-containing film dressing delivered greater than 5.0 log10 reduction throughout the 7 days on all aerobic gram-negative bacilli and Staphylococcus species tested. As of day 1 the CHG-containing film dressing provided greater than 5.0 log10 reduction on Candida albicans. There were no statistically significant differences in the log10 reduction between the two dressings tested. The new CHG-containing film dressing was found to be as effective as the chlorhexidine gluconate-impregnated gel dressing on clinically relevant microbes.

Aerobic Bacteria in the Diaphragmatic Portion of Stethoscope of Medical Professionals of Tertiary Care Hospital.

PubMed

Bham, G; Bhandari, J; Neupane, M R; Dawadi, R; Pradhan, P

2015-01-01

Hospital environment is a reservoir of wide varieties of microorganisms which are frequently reported colonizing in medical equipment. Stethoscopes are essential tools and of universal use in the medical profession, which might be a source of spreading nosocomial infections. This research project was conducted with an aim to assess the presence of aerobic bacteria in the stethoscope of the medical doctors working at Patan Hospital and students of Patan Academy of Health Sciences. It is a cross sectional study based on structured questionnaire and sample assessment from the stethoscope of doctors and students of Patan Hospital and Patan Academy of Health Sciences. The stethescopes used by the doctors of five major departments of Patan Hospital and students of clinical years were included in this study. Total of 99 stethoscope owned by different level of professionals (positions) and different departments were examined for bacterial contamination. Out of them, 36 were found to be considerably contaminated. Single strain of bacteria was grown from a single stethoscope. Among them 34 were Gram positive and remaining were Gram negative. Out of 34 gram postive bacteria, 29 were identified as Staphylococcus aureus, six were identified as Coagulase Negative Staphylococcus and remaining were Gram positive bacilli. There is presence of aerobic bacteria in diaphragmatic portion of stethoscope of medical professional of which the gram positives were the commonest.

Double- and multi-carbapenemase-producers: the excessively armored bacilli of the current decade.

PubMed

Meletis, G; Chatzidimitriou, D; Malisiovas, N

2015-08-01

Multidrug-resistant (MDR) and extensively drug-resistant (XDR) Gram-negative nosocomial pathogens commonly carry one carbapenemase gene conferring resistance to carbapenems and other beta-lactam antibiotics. However, increasing reports show that double-carbapenemase-producing (DCP) and even multi-carbapenemase-producing (MCP) bacteria are emerging in some parts of the world, diminishing further, in some cases, the already limited treatment options. In the present review, the up-to-date reports of DCP and MCP isolates are summarized and concerns regarding their emergence are discussed.

Simplified Protocol for Carba NP Test for Enhanced Detection of Carbapenemase Producers Directly from Bacterial Cultures

PubMed Central

Pasteran, Fernando; Tijet, Nathalie; Melano, Roberto G.

2015-01-01

We compared carbapenemase detection among 266 Gram-negative bacilli (161 carbapenemase producers) using the Carba NP tests issued by the CLSI (CNPt-CLSI) and a novel protocol (CNPt-direct) designed for carbapenemase detection direct from bacterial cultures (instead of bacterial extracts required by the CLSI tests). The specificities were comparable (100%), but the CNPt-direct was more sensitive (98% versus 84%). The CNPt-direct was easier to perform due to the direct use of colonies and offered a more robust detection of carbapenemase producers. PMID:26424841

Epidemic microclusters of blood-culture proven sepsis in very-low-birth weight infants: experience of the German Neonatal Network.

PubMed

Härtel, Christoph; Faust, Kirstin; Avenarius, Stefan; Bohnhorst, Bettina; Emeis, Michael; Gebauer, Corinna; Groneck, Peter; Heitmann, Friedhelm; Hoehn, Thomas; Hubert, Mechthild; Kribs, Angela; Küster, Helmut; Laux, Reinhard; Mögel, Michael; Müller, Dirk; Olbertz, Dirk; Roll, Claudia; Siegel, Jens; Stein, Anja; Vochem, Matthias; Weller, Ursula; von der Wense, Axel; Wieg, Christian; Wintgens, Jürgen; Hemmelmann, Claudia; Simon, Arne; Herting, Egbert; Göpel, Wolfgang

2012-01-01

We evaluated blood culture-proven sepsis episodes occurring in microclusters in very-low-birth-weight infants born in the German Neonatal Network (GNN) during 2009-2010. Thirty-seven centers participated in GNN; 23 centers enrolled ≥50 VLBW infants in the study period. Data quality was approved by on-site monitoring. Microclusters of sepsis were defined as occurrence of at least two blood-culture proven sepsis events in different patients of one center within 3 months with the same bacterial species. For microcluster analysis, we selected sepsis episodes with typically cross-transmitted bacteria of high clinical significance including gram-negative rods and Enterococcus spp. In our cohort, 12/2110 (0.6%) infants were documented with an early-onset sepsis and 235 late-onset sepsis episodes (≥72 h of age) occurred in 203/2110 (9.6%) VLBW infants. In 182/235 (77.4%) late-onset sepsis episodes gram-positive bacteria were documented, while coagulase negative staphylococci were found to be the most predominant pathogens (48.5%, 95%CI: 42.01-55.01). Candida spp. and gram-negative bacilli caused 10/235 (4.3%, 95%CI: 1.68% -6.83%) and 43/235 (18.5%) late-onset sepsis episodes, respectively. Eleven microclusters of blood-culture proven sepsis were detected in 7 hospitals involving a total 26 infants. 16/26 cluster patients suffered from Klebsiella spp. sepsis. The median time interval between the first patient's Klebsiella spp. sepsis and cluster cases was 14.1 days (interquartile range: 1-27 days). First patients in the cluster, their linked cases and sporadic sepsis events did not show significant differences in short term outcome parameters. Microclusters of infection are an important phenomenon for late-onset sepsis. Most gram-negative cluster infections occur within 30 days after the first patient was diagnosed and Klebsiella spp. play a major role. It is essential to monitor epidemic microclusters of sepsis in surveillance networks to adapt clinical practice, inform policy and further improve quality of care.

[Antimicrobial sensitivity of the environmental microbiota in the intensive care units of a peruvian hospital].

PubMed

Díaz-Tello, José; Rojas-Jaimes, Jesús; Ibarra-Trujillo, Jimmy; Tárraga-Gonzales, Delza

2017-01-01

The objective was to detect Gram-negative bacilli and Gram-positive cocci isolated from the environmental microbiota of the Intensive Care Unit (ICU) departments of Neonatology, Pediatrics, and Transplants (kidney, liver, and general) in a Lima hospital and determine their antimicrobial sensitivity. Eighty samples were obtained from inanimate surfaces using a wet swab. A total of 61 bacterial strains were identified, including Staphylococcus epidermis (46.0%), Alcaligenes sp. (21.3%), Pseudomonas aeruginosa (16.4%), Acinetobacter sp. (13.1%), Staphylococcus aureus (1.6%), and Staphylococcus haemolyticus (1.6%). Acinetobacter sp. and P. aeruginosa showed a heightened sensitivity to the antibiotics assessed, while Alcaligenes sp. and S. epidermidis presented the highest antimicrobial resistance. It is recommended that sustained asepsis and monitoring methods be used in ICUs.

[Pneumonia caused byCorynebacterium pseudodiphtheriticum].

PubMed

Furiasse, Daniela; Gasparotto, Ana M; Monterisi, Aída; Castellano, Gabriela; Rocchi, Marta

Microorganisms of the genera Corynebacterium, specie pseudodiphtheriticum are a part of the indigenous microbiota of human skin and oropharinx. Nevertheless in recent decades these bacilli are emerging as opportunistic pathogens causing clinically significant infections in patients with previous compromise. We report the case of a 76 years old female patient, with a history of hypertension, hypothyroidism, type 2 diabetes and chronic renal failure, who presented pneumonia during their stay at the intensive care unit. The induced sputum revealed a representative sample with monomicrobial gram positive pleomorphic coryneform rods (Gram stain) and cultures demonstrated the presence of C. pseudodiphtheriticum as the only bacteria recovered. The pacient received an empirical third generation cephalosporin medication with a succesfull recovery. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

First described case of prosthetic joint infection with Clostridium disporicum.

PubMed

McBride, Joseph A; Sterkel, Alana K; Rehrauer, William M; Smith, Jeannina A

2017-12-01

An orthopedic hardware infection with Clostridium disporicum is described. C. disporicum is a gram positive anaerobic bacillus which can contain two subterminal spores. C. disporicum had not previously been reported in musculoskeletal infections. Gram stains demonstrating gram positive bacilli with two subterminal spores should alert practitioners to the possibility of C. disporicum infection. Published by Elsevier Ltd.

Ceftolozane/tazobactam: a novel cephalosporin/β-lactamase inhibitor combination with activity against multidrug-resistant gram-negative bacilli.

PubMed

Zhanel, George G; Chung, Phillip; Adam, Heather; Zelenitsky, Sheryl; Denisuik, Andrew; Schweizer, Frank; Lagacé-Wiens, Philippe R S; Rubinstein, Ethan; Gin, Alfred S; Walkty, Andrew; Hoban, Daryl J; Lynch, Joseph P; Karlowsky, James A

2014-01-01

Ceftolozane is a novel cephalosporin currently being developed with the β-lactamase inhibitor tazobactam for the treatment of complicated urinary tract infections (cUTIs), complicated intra-abdominal infections (cIAIs), and ventilator-associated bacterial pneumonia (VABP). The chemical structure of ceftolozane is similar to that of ceftazidime, with the exception of a modified side-chain at the 3-position of the cephem nucleus, which confers potent antipseudomonal activity. As a β-lactam, its mechanism of action is the inhibition of penicillin-binding proteins (PBPs). Ceftolozane displays increased activity against Gram-negative bacilli, including those that harbor classical β-lactamases (e.g., TEM-1 and SHV-1), but, similar to other oxyimino-cephalosporins such as ceftazidime and ceftriaxone, it is compromised by extended-spectrum β-lactamases (ESBLs) and carbapenemases. The addition of tazobactam extends the activity of ceftolozane to include most ESBL producers as well as some anaerobic species. Ceftolozane is distinguished from other cephalosporins by its potent activity versus Pseudomonas aeruginosa, including various drug-resistant phenotypes such as carbapenem, piperacillin/tazobactam, and ceftazidime-resistant isolates, as well as those strains that are multidrug-resistant (MDR). Its antipseudomonal activity is attributed to its ability to evade the multitude of resistance mechanisms employed by P. aeruginosa, including efflux pumps, reduced uptake through porins and modification of PBPs. Ceftolozane demonstrates linear pharmacokinetics unaffected by the coadministration of tazobactam; specifically, it follows a two-compartmental model with linear elimination. Following single doses, ranging from 250 to 2,000 mg, over a 1-h intravenous infusion, ceftolozane displays a mean plasma half-life of 2.3 h (range 1.9-2.6 h), a steady-state volume of distribution that ranges from 13.1 to 17.6 L, and a mean clearance of 102.4 mL/min. It demonstrates low plasma protein binding (20 %), is primarily eliminated via urinary excretion (≥92 %), and may require dose adjustments in patients with a creatinine clearance <50 mL/min. Time-kill experiments and animal infection models have demonstrated that the pharmacokinetic-pharmacodynamic index that is best correlated with ceftolozane's in vivo efficacy is the percentage of time in which free plasma drug concentrations exceed the minimum inhibitory concentration of a given pathogen (%fT >MIC), as expected of β-lactams. Two phase II clinical trials have been conducted to evaluate ceftolozane ± tazobactam in the settings of cUTIs and cIAIs. One trial compared ceftolozane 1,000 mg every 8 h (q8h) versus ceftazidime 1,000 mg q8h in the treatment of cUTI, including pyelonephritis, and demonstrated similar microbiologic and clinical outcomes, as well as a similar incidence of adverse effects after 7-10 days of treatment, respectively. A second trial has been conducted comparing ceftolozane/tazobactam 1,000/500 mg and metronidazole 500 mg q8h versus meropenem 1,000 mg q8h in the treatment of cIAI. A number of phase I and phase II studies have reported ceftolozane to possess a good safety and tolerability profile, one that is consistent with that of other cephalosporins. In conclusion, ceftolozane is a new cephalosporin with activity versus MDR organisms including P. aeruginosa. Tazobactam allows the broadening of the spectrum of ceftolozane versus β-lactamase-producing Gram-negative bacilli including ESBLs. Potential roles for ceftolozane/tazobactam include empiric therapy where infection by a resistant Gram-negative organism (e.g., ESBL) is suspected, or as part of combination therapy (e.g., with metronidazole) where a polymicrobial infection is suspected. In addition, ceftolozane/tazobactam may represent alternative therapy to the third-generation cephalosporins after treatment failure or for documented infections due to Gram-negative bacilli producing ESBLs. Finally, the increased activity of ceftolozane/tazobactam versus P. aeruginosa, including MDR strains, may lead to the treatment of suspected and documented P. aeruginosa infections with this agent. Currently, ceftolozane/tazobactam is being evaluated in three phase III trials for the treatment of cUTI, cIAI, and VABP.

Effect of selective decontamination on antimicrobial resistance in intensive care units: a systematic review and meta-analysis.

PubMed

Daneman, Nick; Sarwar, Syed; Fowler, Robert A; Cuthbertson, Brian H

2013-04-01

Many meta-analyses have shown reductions in infection rates and mortality associated with the use of selective digestive decontamination (SDD) or selective oropharyngeal decontamination (SOD) in intensive care units (ICUs). These interventions have not been widely implemented because of concerns that their use could lead to the development of antimicrobial resistance in pathogens. We aimed to assess the effect of SDD and SOD on antimicrobial resistance rates in patients in ICUs. We did a systematic review of the effect of SDD and SOD on the rates of colonisation or infection with antimicrobial-resistant pathogens in patients who were critically ill. We searched for studies using Medline, Embase, and Cochrane databases, with no limits by language, date of publication, study design, or study quality. We included all studies of selective decontamination that involved prophylactic application of topical non-absorbable antimicrobials to the stomach or oropharynx of patients in ICUs, with or without additional systemic antimicrobials. We excluded studies of interventions that used only antiseptic or biocide agents such as chlorhexidine, unless antimicrobials were also included in the regimen. We used the Mantel-Haenszel model with random effects to calculate pooled odds ratios. We analysed 64 unique studies of SDD and SOD in ICUs, of which 47 were randomised controlled trials and 35 included data for the detection of antimicrobial resistance. When comparing data for patients in intervention groups (those who received SDD or SOD) versus data for those in control groups (who received no intervention), we identified no difference in the prevalence of colonisation or infection with Gram-positive antimicrobial-resistant pathogens of interest, including meticillin-resistant Staphylococcus aureus (odds ratio 1·46, 95% CI 0·90-2·37) and vancomycin-resistant enterococci (0·63, 0·39-1·02). Among Gram-negative bacilli, we detected no difference in aminoglycoside-resistance (0·73, 0·51-1·05) or fluoroquinolone-resistance (0·52, 0·16-1·68), but we did detect a reduction in polymyxin-resistant Gram-negative bacilli (0·58, 0·46-0·72) and third-generation cephalosporin-resistant Gram-negative bacilli (0·33, 0·20-0·52) in recipients of selective decontamination compared with those who received no intervention. We detected no relation between the use of SDD or SOD and the development of antimicrobial-resistance in pathogens in patients in the ICU, suggesting that the perceived risk of long-term harm related to selective decontamination cannot be justified by available data. However, our study indicates that the effect of decontamination on ICU-level antimicrobial resistance rates is understudied. We recommend that future research includes a non-crossover, cluster randomised controlled trial to assess long-term ICU-level changes in resistance rates. None. Copyright © 2013 Elsevier Ltd. All rights reserved.

Novel Guanidine Compound against Multidrug-Resistant Cystic Fibrosis-Associated Bacterial Species.

PubMed

Saeed, Aamer; Bosch, Alejandra; Bettiol, Marisa; Nossa González, Diana L; Erben, Mauricio Federico; Lamberti, Yanina

2018-05-11

Chronic pulmonary infection is a hallmark of lung disease in cystic fibrosis (CF). Infections dominated by non-fermentative Gram-negative bacilli are particularly difficult to treat and highlight an urgent need for the development of new class of agents to combat these infections. In this work, a small library comprising thiourea and guanidine derivatives with low molecular weight was designed; these derivatives were studied as antimicrobial agents against Gram-positive, Gram-negative, and a panel of drug-resistant clinical isolates recovered from patients with CF. One novel compound, a guanidine derivative bearing adamantane-1-carbonyl and 2-bromo-4,6-difluouro-phenyl substituents ( H-BDF ), showed potent bactericidal activity against the strains tested, at levels generally higher than those exhibited by tobramycin, ceftazimide and meropenem. The role that different substituents exert in the antimicrobial activity has been determined, highlighting the importance of the halo-phenyl group in the guanidine moiety. The new compound displays low levels of cytotoxicity against THP-1 and A549 cells with a selective index (SI) > 8 (patent application PCT/IB2017/054870, August 2017). Taken together, our results indicate that H-BDF can be considered as a promising antimicrobial agent.

BACTERIAL CONTAMINATION OF STETHOSCOPES

PubMed Central

Bukharie, Huda A.; Al-Zahrani, Hussain; Rubaish, Abdullah M.; Abdulmohsen, Mohammed F.

2004-01-01

Background: A stethoscope, an essential tool of the medical profession, can become a source of nosocomial infection. Objective: To determine the frequency of bacterial contamination of stethoscopes as well as the practices used for cleaning them. Methods: Cultures were taken from 100 stethoscopes used by different medical personnel in different hospital services. The stethoscopes were collected while the staff filled in a questionnaire. Results: Thirty (30%) out of the 100 stethoscopes surveyed were contaminated with microorganisms. The majority of organisms isolated were gram-positive bacteria (gram positive bacilli 12%, gram-negative bacteria 9%, gram-positive cocci 9%). None of the stethoscopes grew methicillin-resistant staphylococcus aureus. Overall, 21% of the health personnel cleaned their stethoscopes daily, 47% weekly, and 32% yearly. None of the health care workers cleaned their stethoscopes after every patient. Nurses cleaned their stethoscopes more often than physicians and medical students. Conclusion: Stethoscopes may be important in the spread of infectious agents. Their regular disinfection after use on each patient should be considered, particularly in such areas of the hospital, as the critical care units, and oncology units which house many patients with antibiotic-resistant organisms. PMID:23012043

Alarming emergence, molecular characterization, and outcome of blaNDM-1 in patients infected with multidrug-resistant Gram-negative bacilli in a tertiary care hospital

PubMed Central

Naim, Huma; Rizvi, Meher; Azam, Mohd; Gupta, Richa; Taneja, Neelam; Shukla, Indu; Khan, Haris M.

2017-01-01

INTRODUCTION: This study was conducted to assess the prevalence of metallo-beta-lactamases (MBLs) in general and blaNDM-1 in particular. It also aimed at evaluating clinical characteristics and outcome in patients infected with MBLs. MATERIALS AND METHODS: A total of 116 carbapenem-resistant Gram-negative bacilli (CRGNB) were evaluated in the study. These CRGNB were tested for MBL production both phenotypically for MBLs and genotypically for blaNDM-1 gene by polymerase chain reaction (PCR). Representative stains of NDM-1 isolates were further sequenced by Triyat Scientific Co., (Nagpur, India). RESULTS: Among 116 CRGNB Citrobacter species 28 (24.13%) was the most common pathogen. Phenotypically, MHT, imipenem-EDTA (IPM-EDTA) double-disk synergy test and IPM-EDTA combined disk synergy test (CDST) detected MBL production in 105 (90.51%), 96 (81.03%), and 87 (75%) CRGNB, respectively. However, blaNDM-1 genes were detected in 66 (56.89%) isolates. The prevalence of blaNDM-1 gene was highest among Escherichia coli 26 (100%). Considering PCR as gold standard, it was observed that IMP-EDTA CDST was most specific (78.38%) while MHT was most sensitive (97.47%). Results of blaNDM-1 gene by PCR were further confirmed by sequencing (Triyat genomics, Nagpur). All the 11 representative strains were confirmed to be an NDM-1 gene. The presence of MBLs in our group of patients (non-Intensive Care Unit patients) is a cause for concern. However, on tracing their outcome, it was interesting to note that while the duration of stay lengthened in a large number of patients 112 (96.5%), mortality was relatively low 5 (4.31%). CONCLUSION: The results of this study provide insight into the prevalence of MBLs, including blaNDM-1, in a tertiary care hospital. Antibiotic stewardship implemented in all seriousness may to a great extent stave off the impending pan-drug resistance. The surprising outcome of our patients suggests either that the bacteria trade off virulence for drug resistance or the relatively robust immune response of non ICU patients fights back. PMID:28706386

Fluoroquinolone antimicrobial agents.

PubMed Central

Wolfson, J S; Hooper, D C

1989-01-01

The fluoroquinolones, a new class of potent orally absorbed antimicrobial agents, are reviewed, considering structure, mechanisms of action and resistance, spectrum, variables affecting activity in vitro, pharmacokinetic properties, clinical efficacy, emergence of resistance, and tolerability. The primary bacterial target is the enzyme deoxyribonucleic acid gyrase. Bacterial resistance occurs by chromosomal mutations altering deoxyribonucleic acid gyrase and decreasing drug permeation. The drugs are bactericidal and potent in vitro against members of the family Enterobacteriaceae, Haemophilus spp., and Neisseria spp., have good activity against Pseudomonas aeruginosa and staphylococci, and (with several exceptions) are less potent against streptococci and have fair to poor activity against anaerobic species. Potency in vitro decreases in the presence of low pH, magnesium ions, or urine but is little affected by different media, increased inoculum, or serum. The effects of the drugs in combination with a beta-lactam or aminoglycoside are often additive, occasionally synergistic, and rarely antagonistic. The agents are orally absorbed, require at most twice-daily dosing, and achieve high concentrations in urine, feces, and kidney and good concentrations in lung, bone, prostate, and other tissues. The drugs are efficacious in treatment of a variety of bacterial infections, including uncomplicated and complicated urinary tract infections, bacterial gastroenteritis, and gonorrhea, and show promise for therapy of prostatitis, respiratory tract infections, osteomyelitis, and cutaneous infections, particularly when caused by aerobic gram-negative bacilli. Fluoroquinolones have also proved to be efficacious for prophylaxis against travelers' diarrhea and infection with gram-negative bacilli in neutropenic patients. The drugs are effective in eliminating carriage of Neisseria meningitidis. Patient tolerability appears acceptable, with gastrointestinal or central nervous system toxicities occurring most commonly, but only rarely necessitating discontinuance of therapy. In 17 of 18 prospective, randomized, double-blind comparisons with another agent or placebo, fluoroquinolones were tolerated as well as or better than the comparison regimen. Bacterial resistance has been uncommonly documented but occurs, most notably with P. aeruginosa and Staphylococcus aureus and occasionally other species for which the therapeutic ratio is less favorable. Fluoroquinolones offer an efficacious, well-tolerated, and cost-effective alternative to parenteral therapies of selected infections. PMID:2680058

Does negative-pressure wound therapy influence subjacent bacterial growth? A systematic review.

PubMed

Glass, Graeme E; Murphy, George R F; Nanchahal, Jagdeep

2017-08-01

Negative-pressure wound therapy is a ubiquitous wound management resource. The influence of NPWT on the bacterial bioburden of the subjacent wound remains unclear. We sought to examine the evidence. MEDLINE, Embase, PubMed, the Cochrane Database of Systematic Reviews and the Cochrane Controlled Trials Register were searched for articles quantitatively evaluating bacterial load under NPWT. Twenty-four studies met the inclusion criteria including 4 randomised controlled trials, 8 clinical series and 12 experimental studies. Twenty studies evaluated conventional NPWT, while 4 evaluated infiltration-based NPWT. While 8 studies using conventional NPWT failed to demonstrate an observable effect on bacterial load, 7 studies reported that NPWT was inherently bacteriostatic and 5 others reported species selectivity with suppression of non-fermentative gram-negative bacilli (NFGNB), including Pseudomonas spp. Simultaneously, there was some evidence of enhanced proliferation of gram-positive cocci where the niche was cleared of NFGNB. Two of the 4 studies using infiltration-based NPWT also reported selectively impaired proliferation of Pseudomonas spp. The assumption that NPWT suppresses bacterial proliferation is oversimplified. There is evidence that NPWT exhibits species selectivity, suppressing the proliferation of NFGNB. However, this may depopulate the niche for exploitation by gram-positive cocci. This, in turn, has implications for the use of NPWT where highly virulent strains of gram-positive cocci have been isolated and the duration of NPWT therapy and frequency of dressing changes. Copyright © 2017 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.

Ready for a world without antibiotics? The Pensières Antibiotic Resistance Call to Action

PubMed Central

2012-01-01

Resistance to antibiotics has increased dramatically over the past few years and has now reached a level that places future patients in real danger. Microorganisms such as Escherichia coli and Klebsiella pneumoniae, which are commensals and pathogens for humans and animals, have become increasingly resistant to third-generation cephalosporins. Moreover, in certain countries, they are also resistant to carbapenems and therefore susceptible only to tigecycline and colistin. Resistance is primarily attributed to the production of beta-lactamase genes located on mobile genetic elements, which facilitate their transfer between different species. In some rare cases, Gram-negative rods are resistant to virtually all known antibiotics. The causes are numerous, but the role of the overuse of antibiotics in both humans and animals is essential, as well as the transmission of these bacteria in both the hospital and the community, notably via the food chain, contaminated hands, and between animals and humans. In addition, there are very few new antibiotics in the pipeline, particularly for Gram-negative bacilli. The situation is slightly better for Gram-positive cocci as some potent and novel antibiotics have been made available in recent years. A strong and coordinated international programme is urgently needed. To meet this challenge, 70 internationally recognized experts met for a two-day meeting in June 2011 in Annecy (France) and endorsed a global call to action ("The Pensières Antibiotic Resistance Call to Action"). Bundles of measures that must be implemented simultaneously and worldwide are presented in this document. In particular, antibiotics, which represent a treasure for humanity, must be protected and considered as a special class of drugs. PMID:22958833

A prediction tool for nosocomial multi-drug Resistant Gram-Negative Bacilli infections in critically ill patients - prospective observational study.

PubMed

Vasudevan, Anupama; Mukhopadhyay, Amartya; Li, Jialiang; Yuen, Eugene Goh Yu; Tambyah, Paul Ananth

2014-11-25

The widespread use of empiric broad spectrum antibiotics has contributed to the global increase of Resistant Gram-Negative Bacilli (RGNB) infections in intensive care units (ICU). The aim of this study was to develop a tool to predict nosocomial RGNB infections among ICU patients for targeted therapy. We conducted a prospective observational study from August'07 to December'11. All adult patients who were admitted and stayed for more than 24 hours at the medical and surgical ICU's were included. All patients who developed nosocomial RGNB infections 48 hours after ICU admission were identified. A prediction score was formulated by using independent risk factors obtained from logistic regression analysis. This was prospectively validated with a subsequent cohort of patients admitted to the ICUs during the following time period of January-September 2012. Seventy-six patients with nosocomial RGNB Infection (31bacteremia) were compared with 1398 patients with Systemic Inflammatory Response Syndrome (SIRS) without any gram negative bacterial infection/colonization admitted to the ICUs during the study period. The following independent risk factors were obtained by a multivariable logistic regression analysis - prior isolation of Gram negative organism (coeff: 1.1, 95% CI 0.5-1.7); Surgery during current admission (coeff: 0.69, 95% CI 0.2-1.2); prior Dialysis with end stage renal disease (coeff: 0.7, 95% CI 0.1-1.1); prior use of Carbapenems (coeff: 1.3, 95% CI 0.3-2.3) and Stay in the ICU for more than 5 days (coeff: 2.4, 95% CI 1.6-3.2). It was validated prospectively in a subsequent cohort (n = 408) and the area-under-the-curve (AUC) of the GSDCS score for predicting nosocomial ICU acquired RGNB infection and bacteremia was 0.77 (95% CI 0.68-0.89 and 0.78 (95% CI 0.69-0.89) respectively. The GSDCS (0-4.3) score clearly differentiated the low (0-1.3), medium (1.4-2.3) and high (2.4-4.3) risk patients, both for RGNB infection (p:0.003) and bacteremia (p:0.009). GSDCS is a simple bedside clinical score which predicts RGNB infection and bacteremia with high predictive value and differentiates low versus high risk patients. This score will help clinicians to choose appropriate, timely targeted antibiotic therapy and avoid exposure to unnecessary treatment for patients at low risk of nosocomial RGNB infection. This will reduce the selection pressure and help to contain antibiotic resistance in ICUs.

In Vitro Activity of Delafloxacin against Contemporary Bacterial Pathogens from the United States and Europe, 2014

PubMed Central

Pfaller, M. A.; Sader, H. S.; Rhomberg, P. R.

2017-01-01

ABSTRACT The in vitro activities of delafloxacin and comparator antimicrobial agents against 6,485 bacterial isolates collected from medical centers in Europe and the United States in 2014 were tested. Delafloxacin was the most potent agent tested against methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus, Streptococcus pneumoniae, viridans group streptococci, and beta-hemolytic streptococci and had activity similar to that of ciprofloxacin and levofloxacin against certain members of the Enterobacteriaceae. Overall, the broadest coverage of the tested pathogens (Gram-positive cocci and Gram-negative bacilli) was observed with meropenem and tigecycline in both Europe and the United States. Delafloxacin was shown to be active against organisms that may be encountered in acute bacterial skin and skin structure infections, respiratory infections, and urinary tract infections. PMID:28167542

In Vitro Activity of Delafloxacin against Contemporary Bacterial Pathogens from the United States and Europe, 2014.

PubMed

Pfaller, M A; Sader, H S; Rhomberg, P R; Flamm, R K

2017-04-01

The in vitro activities of delafloxacin and comparator antimicrobial agents against 6,485 bacterial isolates collected from medical centers in Europe and the United States in 2014 were tested. Delafloxacin was the most potent agent tested against methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus , Streptococcus pneumoniae , viridans group streptococci, and beta-hemolytic streptococci and had activity similar to that of ciprofloxacin and levofloxacin against certain members of the Enterobacteriaceae Overall, the broadest coverage of the tested pathogens (Gram-positive cocci and Gram-negative bacilli) was observed with meropenem and tigecycline in both Europe and the United States. Delafloxacin was shown to be active against organisms that may be encountered in acute bacterial skin and skin structure infections, respiratory infections, and urinary tract infections. Copyright © 2017 Pfaller et al.

Isolation, identification and sensitivity pattern of microorganisms isolated from the urine of pregnant women.

PubMed

Karim, S; Khan, K I

1994-01-01

The present studies were conducted to detect and identify the microorganism from the urine of pregnant women having urinary tract infection. The antibiotic susceptibility of these isolated microorganisms was also determined. The microorganisms found responsible for the infection were bacteria, fungi, yeast and protozoa. Among the bacteria two were identified as Gram-positive cocci i.e. Staphylococcus aureus and S. epidermidis, the remaining two were Gram-negative bacilli which were Escherichia coil and Pseudomonas aeruginosa. The fungus was identified as AspelEillus niger and the yeast like fungus Candida albican. The only protozoan found in some of the urine samples was Trichomonas vaginalis. These isolated and identified microorganisms were more susceptible to Norfloxacin, Velosef, Minocin, Nitrofurantoin, Malidixic acid and Metronidazole whereas antibiotics Penbritin and Cefaloridine were least effective against these microorganisms.

Serum bactericidal activities of moxifloxacin and levofloxacin against aerobic and anaerobic intra-abdominal pathogens.

PubMed

Stein, Gary E; Schooley, Sharon; Tyrrell, Kerin L; Citron, Diane M; Nicolau, David P; Goldstein, Ellie J C

2008-02-01

We studied the serum bactericidal activity (SBA) of moxifloxacin and levofloxacin against common pathogens associated with complicated intra-abdominal infections. Ten healthy volunteers received a single dose of moxifloxacin (400 mg) and levofloxacin (750 mg) and serum samples were collected at 2, 4, 8, 12, and 24h after the dose of each drug. Bactericidal titers in serum over time were determined for aerobic gram-negative bacilli (Escherichia coli, Klebseilla pneumoniae, and Enterobacter cloacae) and anaerobic bacteria (Bacteroides fragilis, Bacteroides thetaiotaomicron, Prevotella bivia, and Finegoldia magna). Both fluoroquinolones provided rapid (2h) attainment and prolonged (24h) SBA (titers > or = 1:8) against each of the aerobic bacilli studied. SBA was observed for at least 12h against B. fragilis strains with MICs < or = 2 microg/ml to moxifloxacin and < or = 4 microg/ml to levofloxacin. Prolonged (12h) SBA (titers > or = 1:2) was also observed against isolates of B. thetaiotaomicron, P. bivia, and F. magna with moxifloxacin < or = MICs 2 microg/ml.

Antimicrobial-resistant Gram-negative bacteria in febrile neutropenic patients with cancer: current epidemiology and clinical impact.

PubMed

Trecarichi, Enrico M; Tumbarello, Mario

2014-04-01

In the recent years, several studies involving cancer patients have demonstrated a clear trend in the epidemiology of bacterial infections showing a shift in the prevalence from Gram-positive to Gram-negative bacteria and the extensive emergence of antimicrobial-resistant strains among Gram-negatives isolated from the blood. The aim of this systematic review was to examine the recent trends in epidemiology and antimicrobial resistance in Gram-negatives recovered from neutropenic cancer patients, with particular emphasis on the impact of antimicrobial resistance on the clinical outcome of severe infections caused by such microorganisms. Overall, from 2007 to date, the rate of Gram-negative bacteria recovery ranged from 24.7 to 75.8% (mean 51.3%) in cancer patient cohorts. Escherichia coli represented the most common species (mean frequency of isolation 32.1%) among the Gram-negatives, followed by Pseudomonas aeruginosa (mean frequency of isolation 20.1%). An increasing frequency of Acinetobacter spp. and Stenotrophomonas maltophilia was also reported. Increased rates of multidrug-resistant Gram-negative strains have been highlighted among Enterobacteriaceae and nonfermenting Gram-negative rods, despite discontinuation of fluoroquinolone-based antibacterial prophylaxis for neutropenic patients. In addition, antimicrobial resistance and/or the inadequacy of empirical antibiotic treatment have been frequently linked to a worse outcome in cancer patients with bloodstream infections caused by Gram-negative isolates. Sound knowledge of the local distribution of pathogens and their susceptibility patterns and prompt initiation of effective antimicrobial treatment for severe infections caused by Gram-negative bacteria are essential in cancer patients.

Simplified Protocol for Carba NP Test for Enhanced Detection of Carbapenemase Producers Directly from Bacterial Cultures.

PubMed

Pasteran, Fernando; Tijet, Nathalie; Melano, Roberto G; Corso, Alejandra

2015-12-01

We compared carbapenemase detection among 266 Gram-negative bacilli (161 carbapenemase producers) using the Carba NP tests issued by the CLSI (CNPt-CLSI) and a novel protocol (CNPt-direct) designed for carbapenemase detection direct from bacterial cultures (instead of bacterial extracts required by the CLSI tests). The specificities were comparable (100%), but the CNPt-direct was more sensitive (98% versus 84%). The CNPt-direct was easier to perform due to the direct use of colonies and offered a more robust detection of carbapenemase producers. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Rapid Identification of OXA-48 and OXA-163 Subfamilies in Carbapenem-Resistant Gram-Negative Bacilli with a Novel Immunochromatographic Lateral Flow Assay.

PubMed

Pasteran, Fernando; Denorme, Laurence; Ote, Isabelle; Gomez, Sonia; De Belder, Denise; Glupczynski, Youri; Bogaerts, Pierre; Ghiglione, Barbara; Power, Pablo; Mertens, Pascal; Corso, Alejandra

2016-11-01

We assessed a novel immunochromatographic lateral flow assay for direct identification of OXA-48-like carbapenemases and accurate differentiation of allele variants with distinct substrate profiles (OXA-48 or OXA-163 subfamilies). The assay allowed rapid (less than 4 min) and reliable direct confirmation of OXA-163- and/or OXA-48-like enzymes (with 100% sensitivity and 100% specificity) from cultured colonies that were recovered from both solid medium and spiked blood culture bottles. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Comparative Evaluation of Multiplex PCR and Routine Laboratory Phenotypic Methods for Detection of Carbapenemases among Gram Negative Bacilli.

PubMed

Solanki, Rachana; Vanjari, Lavanya; Subramanian, Sreevidya; B, Aparna; E, Nagapriyanka; Lakshmi, Vemu

2014-12-01

Carbapenem resistant pathogens cause infections associated with significant morbidity and mortality. This study evaluates the use of Multiplex PCR for rapid detection of carbapenemase genes among carbapenem resistant Gram negative bacteria in comparison with the existing phenotypic methods like modified Hodge test (MHT), combined disc test (CDT) and automated methods. A total of 100 Carbapenem resistant clinical isolates, [Escherichia coli (25), Klebsiella pneumoniae (35) P. aeruginosa (18) and Acinetobacter baumannii (22)] were screened for the presence of carbapenemases (bla NDM-1, bla VIM , blaIMP and blaKPC genes) by phenotype methods such as the modified Hodge test (MHT) and combined disc test (CDT) and the molecular methods such as Multiplex PCR. Seventy of the 100 isolates were MHT positive while, 65 isolates were positive by CDT. All the CDT positive isolates with EDTA and APB were Metallo betalactamase (MBL) and K. pneumoniae carbapenemase (KPC) producers respectively. bla NDM-1 was present as a lone gene in 44 isolates. In 14 isolates bla NDM-1 gene was present with blaKPC gene, and in one isolate bla NDM-1 gene was present with blaVIM , gene. Only one E. coli isolate had a lone blaKPC gene. We didn't find bla IMP gene in any of the isolates. Neither of the genes could be detected in 35 isolates. Accurate detection of the genes related with carbapenemase production by Molecular methods like Multiplex PCR overcome the limitations of the phenotypic methods and Automated systems.

Persistent bacteremia secondary to delayed identification of Lactobacillus in the setting of mitral valve endocarditis.

PubMed

Stroupe, Cody; Pendley, Joseph; Isang, Emmanuel; Helms, Benjamin

2017-01-01

Lactobacillus species causing infective endocarditis is rare. Most reported cases arise from the oral ingestion of Lactobacillus via dairy or nutritional supplements in patients with congenital valve disease or replacement. We present a case of native valve bacterial endocarditis caused by Lactobacillus arising from dental abscesses. Additionally, there was an error in identification of the Lactobacillus as Corynebacterium , which led to inadequate treatment. A 51-year-old male presented to an outside clinic with several weeks of subjective fevers and malaise. The provider obtained two sets of blood cultures. Both grew Gram-positive bacilli identified as Corynebacterium . Once hospitalized he persistently had positive blood cultures despite treatment with vancomycin and gentamicin. The specimens were sent to a reference lab. The cultures were confirmed to be Lactobacillus zeae resistant to vancomycin and gentamicin. Once he was started on appropriate therapy his blood cultures showed no further growth of bacteria. The infected teeth were removed as it was felt they were the source of the bacteremia. This case presents two interesting topics in one encounter. First, Lactobacillus is not a common culprit in endocarditis. Secondly, the incorrect identification of the gram-positive bacilli bacteria led to prolonged bacteremia in our patient. The patient was evaluated by cardiothoracic surgery at our facility and it was determined that he would likely need a mitral valve replacement versus repair. The decision was made to treat the patient with six weeks Penicillin-VK prior to the operation. He is currently completing his antibiotic therapy.

Direct detection of Streptococcus mutans in human dental plaque by polymerase chain reaction.

PubMed

Igarashi, T; Yamamoto, A; Goto, N

1996-10-01

Streptococcus mutans is an etiological agent in human dental caries. A method for the detection of S. mutans directly from human dental plaque by polymerase chain reaction has been developed. Oligonucleotide primers specific for a portion of the dextranase gene (dexA) of S. mutans Ingbritt (serotype c) were designed to amplify a 1272-bp DNA fragment by polymerase chain reaction. The present method specifically detected S. mutans (serotypes c, e and f), but none of the other mutans streptococci: S. cricetus (serotype a), S. rattus (serotype b), S. sobrinus (serotypes d and g), and S. downei (serotype h), other gram-positive bacteria (16 strains of 12 species of cocci and 18 strains of 12 species of bacilli) nor gram-negative bacteria (1 strain of 1 species of cocci and 20 strains of 18 species of bacilli). The method was capable of detecting 1 pg of the chromosomal DNA purified from S. mutans Ingbritt and as few as 12 colony-forming units of S. mutans cells. The S. mutans cells in human dental plaque were also directly detected. Seventy clinical isolates of S. mutans isolated from the dental plaque of 8 patients were all positive by the polymerase chain reaction. These results suggest that the dexA polymerase chain reaction is suitable for the specific detection and identification of S. mutans.

Nested polymerase chain reaction (PCR) targeting 16S rDNA for bacterial identification in empyema.

PubMed

Prasad, Rajniti; Kumari, Chhaya; Das, B K; Nath, Gopal

2014-05-01

Empyema in children causes significant morbidity and mortality. However, identification of organisms is a major concern. To detect bacterial pathogens in pus specimens of children with empyema by 16S rDNA nested polymerase chain reaction (PCR) and correlate it with culture and sensitivity. Sixty-six children admitted to the paediatric ward with a diagnosis of empyema were enrolled prospectively. Aspirated pus was subjected to cytochemical examination, culture and sensitivity, and nested PCR targeting 16S rDNA using a universal eubacterial primer. Mean (SD) age was 5·8 (1·8) years (range 1-13). Analysis of aspirated pus demonstrated total leucocyte count >1000×10(6)/L, elevated protein (≧20 g/L) and decreased glucose (≤2·2 mmol/L) in 80·3%, 98·5% and 100%, respectively. Gram-positive cocci were detected in 29 (43·9%) and Gram-negative bacilli in two patients. Nested PCR for the presence of bacterial pathogens was positive in 50·0%, compared with 36·3% for culture. 16S rDNA PCR improves rates of detection of bacteria in pleural fluid, and can detect bacterial species in a single assay as well as identifying unusual and unexpected causal agents.

Pure neuritic leprosy: Resolving diagnostic issues in acid fast bacilli (AFB)-negative nerve biopsies: A single centre experience from South India

PubMed Central

Hui, Monalisa; Uppin, Megha S.; Challa, Sundaram; Meena, A. K.; Kaul, Subhash

2015-01-01

Background and Purpose: Demonstration of lepra bacilli is essential for definite or unequivocal diagnosis of pure neuritic leprosy (PNL) on nerve biopsy. However, nerves always do not show bacilli owing to the changes of previous therapy or due to low bacillary load in tuberculoid forms. In absence of granuloma or lepra bacilli, other morphologic changes in endoneurium and perineurium can be of help in making a probable diagnosis of PNL and treating the patient with multidrug therapy. Materials and Methods: Forty-six biopsies of PNL were retrospectively reviewed and histologic findings were compared with 25 biopsies of non leprosy neuropathies (NLN) including vasculitic neuropathy and chronic inflammatory demyelinating polyneuropathy (CIDP). The distribution of endoneurial infiltrate and fibrosis, perineurial thickening, and myelin abnormalities were compared between PNL and NLN biopsies and analyzed by Chi-square test. Results: Out of 46 PNL casses, 24 (52.17 %) biopsies were negative for acid fast bacilli (AFB). In these cases, the features which favor a diagnosis of AFB-negative PNL were endoneurial infiltrate (51.1%), endoneurial fibrosis (54.2%), perineurial thickening (70.8%), and reduced number of myelinated nerve fibers (75%). Interpretation and Conclusion: Nerve biopsy is an efficient tool to diagnose PNL and differentiate it from other causes of NLN. In absence of AFB, the diagnosis of PNL is challenging. In this article, we have satisfactorily evaluated the various hisopthological features and found that endoneurial inflammation, dense fibrosis, and reduction in the number of myelinated nerve fibers are strong supportive indicators of PNL regardless of AFB positivity. PMID:26425006

Pure neuritic leprosy: Resolving diagnostic issues in acid fast bacilli (AFB)-negative nerve biopsies: A single centre experience from South India.

PubMed

Hui, Monalisa; Uppin, Megha S; Challa, Sundaram; Meena, A K; Kaul, Subhash

2015-01-01

Demonstration of lepra bacilli is essential for definite or unequivocal diagnosis of pure neuritic leprosy (PNL) on nerve biopsy. However, nerves always do not show bacilli owing to the changes of previous therapy or due to low bacillary load in tuberculoid forms. In absence of granuloma or lepra bacilli, other morphologic changes in endoneurium and perineurium can be of help in making a probable diagnosis of PNL and treating the patient with multidrug therapy. Forty-six biopsies of PNL were retrospectively reviewed and histologic findings were compared with 25 biopsies of non leprosy neuropathies (NLN) including vasculitic neuropathy and chronic inflammatory demyelinating polyneuropathy (CIDP). The distribution of endoneurial infiltrate and fibrosis, perineurial thickening, and myelin abnormalities were compared between PNL and NLN biopsies and analyzed by Chi-square test. Out of 46 PNL casses, 24 (52.17 %) biopsies were negative for acid fast bacilli (AFB). In these cases, the features which favor a diagnosis of AFB-negative PNL were endoneurial infiltrate (51.1%), endoneurial fibrosis (54.2%), perineurial thickening (70.8%), and reduced number of myelinated nerve fibers (75%). Nerve biopsy is an efficient tool to diagnose PNL and differentiate it from other causes of NLN. In absence of AFB, the diagnosis of PNL is challenging. In this article, we have satisfactorily evaluated the various hisopthological features and found that endoneurial inflammation, dense fibrosis, and reduction in the number of myelinated nerve fibers are strong supportive indicators of PNL regardless of AFB positivity.

The diagnosis and treatment of donovanosis (granuloma inguinale).

PubMed Central

Richens, J

1991-01-01

Donovanosis is a predominantly tropical cause of genital ulcer occurring chiefly in small endemic foci in all continents except Europe. Diagnosis requires the careful collection, staining and examination of smears or biopsies of characteristic genital and, occasionally, extragenital lesions for demonstration of the pathognomonic Donovan bodies (Calymmatobacterium granulomatis) within histiocytes. Successful isolation of C. granulomatis has rarely proved feasible, the last report being in 1962. Donovanosis has a characteristic histopathological picture which occasionally simulates epithelioma. The antibiotics reported as showing good activity in donovanosis are those with good activity against gram negative bacilli and whose lipid solubility ensures good intracellular penetration. They include streptomycin, chloramphenicol, erythromycin, lincomycin, cotrimoxazole and the tetracyclines. More recently, good results have been reported with norfloxacin and thiamphenicol. The treatment of donovanosis in pregnant women and patients with AIDS poses special problems. Complications of donovanosis such as elephantiasis, stricture and pelvic abscess may require surgery. Contacts should be traced for examination but only treated if lesions are found. Images PMID:1774048

Antimicrobial Resistance in Asia: Current Epidemiology and Clinical Implications

PubMed Central

Kang, Cheol-In

2013-01-01

Antimicrobial resistance has become one of the most serious public health concerns worldwide. Although circumstances may vary by region or country, it is clear that some Asian countries are epicenters of resistance, having seen rapid increases in the prevalence of antimicrobial resistance of major bacterial pathogens. In these locations, however, the public health infrastructure to combat this problem is very poor. The prevalence rates of methicillin-resistant Staphylococcus aureus (MRSA), macrolide-resistant Streptococcus pneumoniae, and multidrug-resistant enteric pathogens are very high due to the recent emergence of extremely drug-resistant gram-negative bacilli in Asia. Because antimicrobial options for these pathogens are extremely limited, infections caused by antimicrobial-resistant bacteria are often associated with inappropriate antimicrobial therapy and poor clinical outcomes. Physicians should be aware of the current epidemiological status of resistance and understand the appropriate use of antimicrobial agents in clinical practice. This review focuses on describing the epidemiology and clinical implications of antimicrobial-resistant bacterial infections in Asian countries. PMID:24265947

EVALUATION OF OCULAR PROSTHESIS BIOFILM AND ANOPHTHALMIC CAVITY CONTAMINATION AFTER USE OF THREE CLEANSING SOLUTIONS

PubMed Central

Paranhos, Regina Márcia Zuccolotto Felippe; Batalhão, Carlos Henrique; Semprini, Marisa; Regalo, Simone Cecílio Hallak; Ito, Izabel Yoko; de Mattos, Maria da Glória Chiarello

2007-01-01

In addition to an initial socket discomfort, ocular prosthesis (OP) installation may allow the adherence of fungi and/or bacteria due to the superficial characteristics of the prosthesis’ material, use of inadequate cleansing solutions and methods, or because the void located between the internal portion of the prosthesis and the anophthalmic cavity (AC) mucosa. Objective: The aim of this study was to evaluate OP biofilm formation and the level of contamination of the internal portion of the OP and the AC in 24 patients. Material and Methods: Material was collected from the AC at the beginning of the study and 15 days after cleansing of the OP with 3 cleansing solutions: a neutral liquid soap, a multiuse solution for contact lens (Complete) and 0.12% chlorhexidine (Periogard). The collected materials were sowed in Petri dishes containing selective media for aerobic and facultative microorganisms, specifically staphylococci (Hipersalt agar with egg yolk), aerobic microorganisms (Brain Heart Infusion Blood Agar), streptococci (Mitis salivarius Agar), gram-negative bacilli (MacConkey Agar) and yeasts (Chromagar CandidaTM), incubated at 35°C or 37°C and the number of colony forming units were counted. Data were analyzed statistically by ANOVA, Friedman’s test and Spearman’s correlation. Results: Aerobic microorganisms, gram-negative bacilli and S. aureus were found in the OP biofilm and in the AC. There was statistically significant difference (p<0.05) between the number of microorganisms before and after the use of the cleansing solutions. Conclusion: There was positive correlation with respect to the microorganisms present in the OP biofilm and AC for the 4 proposed treatments, indicating that the decrease of OP contamination leads to AC contamination as well. PMID:19089097

Towards a phenotypic screening strategy for emerging β-lactamases in Gram-negative bacilli.

PubMed

Willems, Elise; Verhaegen, Jan; Magerman, Koen; Nys, Sita; Cartuyvels, Reinoud

2013-02-01

The purpose of this manuscript was to review recent literature and guidelines regarding phenotypic detection of emerging β-lactamases [extended-spectrum β-lactamases (ESBLs), AmpC β-lactamases and carbapenemases] in Gram-negative bacilli (GNB) in order to formulate recommendations on best practice to screen for them. We conclude that chromogenic ESBL screening agar plates are suitable to screen for ESBL-producing Enterobacteriaceae directly from clinical samples. Furthermore, ceftazidime (CAZ) and ceftriaxone or cefotaxime (CTX) are the indicator antimicrobial agents of choice for ESBL detection in GNB. In non-inducible Enterobacteriaceae, the combined double-disk synergy test (CDDST) with at least CTX and CAZ and additionally cefepime as indicators is the preferred ESBL confirmation assay. The two most suitable ESBL confirmation strategies in AmpC co-producing Enterobacteriaceae are adapted CDDSTs: (i) with addition of 3-aminophenylboronic acid to CTX and CAZ disks; and (ii) with addition of cloxacillin (CLOX) to Mueller-Hinton agar. Reduced cefoxitin susceptibility and decreased susceptibility to cefotetan are regarded as suitable screening tests for plasmid-mediated and derepressed AmpC production. A CLOX-based CDDST with CTX and CAZ as indicators is considered to be the best AmpC confirmation assay. Finally, in Enterobacteriaceae isolates we suggest to screen for carbapenemases with a 0.5 μg/mL meropenem screening breakpoint. For class A carbapenemase confirmation, the home-prepared as well as the commercially available boronic acid-based CDDST can be considered. For metallo-β-lactamase confirmation, ethylene diamine tetra-acetic-acid-based home-prepared assays are recommended. The most suitable method (CDDST or DDST) and indicator antimicrobial agent(s) vary depending on the bacterial genus. Copyright © 2012 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

Effect of Surotomycin, a Novel Cyclic Lipopeptide Antibiotic, on Intestinal Colonization with Vancomycin-Resistant Enterococci and Klebsiella pneumoniae in Mice

PubMed Central

Deshpande, Abhishek; Hurless, Kelly; Cadnum, Jennifer L.; Chesnel, Laurent; Gao, Lihong; Chan, Luisa; Kundrapu, Sirisha; Polinkovsky, Alexander

2016-01-01

Surotomycin (formerly called CB-183,315) is a novel, orally administered cyclic lipopeptide antibacterial in development for the treatment of Clostridium difficile infection (CDI) that has potent activity against vancomycin-resistant enterococci (VRE) but limited activity against Gram-negative bacilli, including Bacteroides spp. We used a mouse model to investigate the impact of surotomycin exposure on the microbiome, and to test the consequences of the disruption on colonization by vancomycin-resistant enterococci (VRE) and extended-spectrum β-lactamase-producing Klebsiella pneumoniae (ESBL-KP), in comparison with the effects of oral vancomycin and metronidazole. Mice (8 per group) received saline, vancomycin, metronidazole, or surotomycin through an orogastric tube daily for 5 days and were challenged with 105 CFU of VRE or ESBL-KP administered through an orogastric tube on day 2 of treatment. The concentrations of the pathogens in stool were determined during and after treatment by plating on selective media. A second experiment was conducted to determine if the antibiotics would inhibit established VRE colonization. In comparison to controls, oral vancomycin promoted VRE and ESBL-KP overgrowth in stool (8 log10 to 10 log10 CFU/g; P < 0.001), whereas metronidazole did not (<4 log10 CFU/g; P > 0.5). Surotomycin promoted ESBL-KP overgrowth (>8 log10 CFU/g; P, <0.001 for comparison with saline controls) but not VRE overgrowth. Surotomycin suppressed preexisting VRE colonization, whereas metronidazole and vancomycin did not. These results suggest that treatment of CDI with surotomycin could reduce levels of VRE acquisition and overgrowth from those with agents such as vancomycin and metronidazole. However, surotomycin and vancomycin may promote colonization by antibiotic-resistant Gram-negative bacilli. PMID:26976870

Emergence of Imipenem-Resistant Gram-Negative Bacilli in Intestinal Flora of Intensive Care Patients

PubMed Central

Angebault, Cécile; Barbier, François; Hamelet, Emilie; Defrance, Gilles; Ruppé, Etienne; Bronchard, Régis; Lepeule, Raphaël; Lucet, Jean-Christophe; El Mniai, Assiya; Wolff, Michel; Montravers, Philippe; Plésiat, Patrick; Andremont, Antoine

2013-01-01

Intestinal flora contains a reservoir of Gram-negative bacilli (GNB) resistant to cephalosporins, which are potentially pathogenic for intensive care unit (ICU) patients; this has led to increasing use of carbapenems. The emergence of carbapenem resistance is a major concern for ICUs. Therefore, in this study, we aimed to assess the intestinal carriage of imipenem-resistant GNB (IR-GNB) in intensive care patients. For 6 months, 523 consecutive ICU patients were screened for rectal IR-GNB colonization upon admission and weekly thereafter. The phenotypes and genotypes of all isolates were determined, and a case control study was performed to identify risk factors for colonization. The IR-GNB colonization rate increased regularly from 5.6% after 1 week to 58.6% after 6 weeks in the ICU. In all, 56 IR-GNB strains were collected from 50 patients: 36 Pseudomonas aeruginosa strains, 12 Stenotrophomonas maltophilia strains, 6 Enterobacteriaceae strains, and 2 Acinetobacter baumannii strains. In P. aeruginosa, imipenem resistance was due to chromosomally encoded resistance (32 strains) or carbapenemase production (4 strains). In the Enterobacteriaceae strains, resistance was due to AmpC cephalosporinase and/or extended-spectrum β-lactamase production with porin loss. Genomic comparison showed that the strains were highly diverse, with 8 exceptions (4 VIM-2 carbapenemase-producing P. aeruginosa strains, 2 Klebsiella pneumoniae strains, and 2 S. maltophilia strains). The main risk factor for IR-GNB colonization was prior imipenem exposure. The odds ratio for colonization was already as high as 5.9 (95% confidence interval [95% CI], 1.5 to 25.7) after 1 to 3 days of exposure and increased to 7.8 (95% CI, 2.4 to 29.8) thereafter. In conclusion, even brief exposure to imipenem is a major risk factor for IR-GNB carriage. PMID:23318796

Multiplex identification of sepsis-causing Gram-negative pathogens from the plasma of infected blood.

PubMed

Chung, Boram; Park, Chulmin; Cho, Sung-Yeon; Shin, Juyoun; Shin, Sun; Yim, Seon-Hee; Lee, Dong-Gun; Chung, Yeun-Jung

2018-02-01

Early and accurate detection of bacterial pathogens in the blood is the most crucial step for sepsis management. Gram-negative bacteria are the most common organisms causing severe sepsis and responsible for high morbidity and mortality. We aimed to develop a method for rapid multiplex identification of clinically important Gram-negative pathogens and also validated whether our system can identify Gram-negative pathogens with the cell-free plasm DNA from infected blood. We designed five MLPA probe sets targeting the genes specific to major Gram-negative pathogens (uidA and lacY for E. coli, ompA for A. baumannii, phoE for K. pneumoniae, and ecfX for P. aeruginosa) and one set targeting the CTX-M group 1 to identify the ESBL producing Gram-negative pathogens. All six target-specific peaks were clearly separated without any non-specific peaks in a multiplex reaction condition. The minimum detection limit was 100 fg of pathogen DNA. When we tested 28 Gram-negative clinical isolates, all of them were successfully identified without any non-specific peaks. To evaluate the clinical applicability, we tested seven blood samples from febrile patients. Three blood culture positive cases showed E. coli specific peaks, while no peak was detected in the other four culture negative samples. This technology can be useful for detection of major sepsis-causing, drug-resistant Gram-negative pathogens and also the major ESBL producing Gram-negatives from the blood of sepsis patients in a clinical setting. This system can help early initiation of effective antimicrobial treatment against Gram-negative pathogens for sepsis patients, which is very crucial for better treatment outcomes. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Bacteremic complications of intravascular catheter tip colonization with Gram-negative micro-organisms in patients without preceding bacteremia.

PubMed

van Eck van der Sluijs, A; Oosterheert, J J; Ekkelenkamp, M B; Hoepelman, I M; Peters, Edgar J G

2012-06-01

Although Gram-negative micro-organisms are frequently associated with catheter-related bloodstream infections, the prognostic value and clinical implication of a positive catheter tip culture with Gram-negative micro-organisms without preceding bacteremia remains unclear. We determined the outcomes of patients with intravascular catheters colonized with these micro-organisms, without preceding positive blood cultures, and identified risk factors for the development of subsequent Gram-negative bacteremia. All patients with positive intravascular catheter tip cultures with Gram-negative micro-organisms at the University Medical Center, Utrecht, The Netherlands, between 2005 and 2009, were retrospectively studied. Patients with Gram-negative bacteremia within 48 h before catheter removal were excluded. The main outcome measure was bacteremia with Gram-negative micro-organisms. Other endpoints were length of the hospital stay, in-hospital mortality, secondary complications of Gram-negative bacteremia, and duration of intensive care admission. A total of 280 catheters from 248 patients were colonized with Gram-negative micro-organisms. Sixty-seven cases were excluded because of preceding positive blood cultures, leaving 213 catheter tips from 181 patients for analysis. In 40 (19%) cases, subsequent Gram-negative bacteremia developed. In multivariate analysis, arterial catheters were independently associated with subsequent Gram-negative bacteremia (odds ratio [OR] = 5.00, 95% confidence interval [CI]: 1.20-20.92), as was selective decontamination of the digestive tract (SDD) (OR = 2.47, 95% CI: 1.07-5.69). Gram-negative bacteremia in patients who received SDD was predominantly caused by cefotaxime (part of the SDD)-resistant organisms. Mortality was significantly higher in the group with subsequent Gram-negative bacteremia (35% versus 20%, OR = 2.12, 95% CI: 1.00-4.49). Patients with a catheter tip colonized with Gram-negative micro-organisms had a high chance of subsequent Gram-negative bacteremia from any cause. This may be clinically relevant, as starting antibiotic treatment pre-emptively in high-risk patients with Gram-negative micro-organisms cultured from arterial intravenous catheters may be beneficial.

Late-onset neonatal sepsis: recent developments.

PubMed

Dong, Ying; Speer, Christian P

2015-05-01

The incidence of neonatal late-onset sepsis (LOS) is inversely related to the degree of maturity and varies geographically from 0.61% to 14.2% among hospitalised newborns. Epidemiological data on very low birth weight infants shows that the predominant pathogens of neonatal LOS are coagulase-negative staphylococci, followed by Gram-negative bacilli and fungi. Due to the difficulties in a prompt diagnosis of LOS and LOS-associated high risk of mortality and long-term neurodevelopmental sequelae, empirical antibiotic treatment is initiated on suspicion of LOS. However, empirical therapy is often inappropriately used with unnecessary broad-spectrum antibiotics and a prolonged duration of treatment. The increasing number of multidrug-resistant Gram-negative micro-organisms in neonatal intensive care units (NICU) worldwide is a serious concern, which requires thorough and efficient surveillance strategies and appropriate treatment regimens. Immunological strategies for preventing neonatal LOS are not supported by current evidence, and approaches, such as a strict hygiene protocol and the minimisation of invasive procedures in NICUs represent the cornerstone to reduce the burden of neonatal LOS. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Reduction in clinical response to empiric antimicrobial therapy of febrile granulocytopenic patients receiving TMP/SMX infection prophylaxis

PubMed Central

Bow, Eric J; Pater, Joseph L; Louie, Thomas J; Feld, Ronald; Mandell, Lionel; Robson, Hugh G; Chow, Anthony; Belch, Andrew; Miedzinski, Lilly; Paul, Nancy; Elliott, Catherine R; Willan, Andrew R

1992-01-01

In the course of a multicentre clinical trial evaluating two antibacterial regimens for the empiric treatment of suspected infection in febrile neutropenic cancer patients, a suboptimal response was noted among recipients of antibacterial prophylaxis with trimethoprim/sulphamethoxazole (TMP/SMX). Multivariate analysis identified TMP/SMX prophylaxis as a predictor of poor outcome independent of other variables such as classification of infection, marrow recovery, neutrophil count at first fever, indwelling central venous catheter use, and underlying disease. This effect appeared to be restricted to recipients of tobramycin plus ticarcillin (TT). TMP/SMX suppresses potentially pathogenic aerobic Gram-negative bacilli and allows colonization and subsequent infection by Gram-positive microorganisms against which TT-like regimens have limited activity. Recognition of this phenomenon may permit a more appropriate selection of antibacterial agents for the therapy of suspected infection in the neutropenic patient. PMID:22416197

The Causes of Post-Operative Meningitis: The Comparison Of Gram-Negative and Gram-Positive Pathogens.

PubMed

Kurtaran, Behice; Kuscu, Ferit; Ulu, Aslihan; Inal, Ayse Seza; Komur, Suheyla; Kibar, Filiz; Cetinalp, Nuri Eralp; Ozsoy, Kerem Mazhar; Arslan, Yusuf Kemal; Aksu, Hasan Salih; Tasova, Yesim

2017-06-20

In this study, we aim to determine the microbiological etiology in critically ill neurosurgical patients with nosocomial meningitis (NM) and show the impact of Gram-negative rods and differences of patient's characteristics, clinical and prognostic measures between Gram-negative and Gram-positive meningitis. In this prospective, one center study we reviewed all adult patients hospitalized during a 12-year period and identified pathogens isolated from post-neurosurgical cases of NM. Demographic, clinical, and treatment characteristics were noted from the medical records. Of the 134 bacterial NM patients, 78 were male and 56 were female, with a mean age of 46±15.9 and median age of 50 (18-80) years. 141 strains isolated; 82 (58.2%) were Gram negative, 59 (41.8%) were Gram positive. Most common isolated microorganism was Acinetobacter baumannii (%34.8). In comparison of mortality data shows that the patients who have meningitis with Gram-negative pathogens have higher mortality than with Gram positives (p=0.034). The duration between surgery and meningitis was shorter in Gram negative meningitis cases compared to others (p=0.045) but the duration between the diagnosis and death was shorter in Gram-positive meningitis cases compared to Gram negatives (p= 0.017). CSF protein and lactate level were higher and glucose level was lower in cases of NM with Gram negatives (p value were respectively, 0.022, 0.039 and 0.049). As conclusions; in NM, Gram-negative pathogens were seen more frequently; A.baumanni was the predominant pathogen; and NM caused by Gram negatives had worse clinical and laboratory characteristic and prognostic outcome than Gram positives.

Bactericidal Effects and Mechanism of Action of Olanexidine Gluconate, a New Antiseptic

PubMed Central

Iwata, Koushi; Nii, Takuya; Nakata, Hikaru; Tsubotani, Yoshie; Inoue, Yasuhide

2015-01-01

Olanexidine gluconate [1-(3,4-dichlorobenzyl)-5-octylbiguanide gluconate] (development code OPB-2045G) is a new monobiguanide compound with bactericidal activity. In this study, we assessed its spectrum of bactericidal activity and mechanism of action. The minimal bactericidal concentrations of the compound for 30-, 60-, and 180-s exposures were determined with the microdilution method using a neutralizer against 320 bacterial strains from culture collections and clinical isolates. Based on the results, the estimated bactericidal olanexidine concentrations with 180-s exposures were 869 μg/ml for Gram-positive cocci (155 strains), 109 μg/ml for Gram-positive bacilli (29 strains), and 434 μg/ml for Gram-negative bacteria (136 strains). Olanexidine was active against a wide range of bacteria, especially Gram-positive cocci, including methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci, and had a spectrum of bactericidal activity comparable to that of commercial antiseptics, such as chlorhexidine and povidone-iodine. In vitro experiments exploring its mechanism of action indicated that olanexidine (i) interacts with the bacterial surface molecules, such as lipopolysaccharide and lipoteichoic acid, (ii) disrupts the cell membranes of liposomes, which are artificial bacterial membrane models, (iii) enhances the membrane permeability of Escherichia coli, (iv) disrupts the membrane integrity of S. aureus, and (v) denatures proteins at relatively high concentrations (≥160 μg/ml). These results indicate that olanexidine probably binds to the cell membrane, disrupts membrane integrity, and its bacteriostatic and bactericidal effects are caused by irreversible leakage of intracellular components. At relatively high concentrations, olanexidine aggregates cells by denaturing proteins. This mechanism differs slightly from that of a similar biguanide compound, chlorhexidine. PMID:25987609

Antimicrobial Activity of a Novel Vascular Access Film Dressing Containing Chlorhexidine Gluconate

PubMed Central

Wibaux, Anne; Thota, Priyaleela; Mastej, Jozef; Prince, Daniel L.; Carty, Neal; Johnson, Peter

2015-01-01

Background Covering insertion sites with chlorhexidine impregnated dressings has been proven to be clinically effective in reducing catheter related blood stream infections (CR-BSI). Two chlorhexidine gluconate (CHG)-impregnated dressings are commercially available, a polyurethane foam disk and a film dressing containing a chlorhexidine gluconate-impregnated gel pad. While both have demonstrated efficacy in clinical settings, the major drawback of high cost and impaired IV insertion site visibility limits their usage. A new, simple film dressing containing CHG within its adhesive layer is now available. The objective of this study was to test the in vitro antimicrobial efficacy of the new dressing in comparison to the CHG-impregnated gel dressing. Methods Quantitative aliquots of suspensions (concentration of 1.0x106 to 5.0x106 cfu/sample) of clinically relevant challenge organisms (Staphylococcus species, gram-negative bacilli, Candida albicans) were incubated in contact with the new CHG-containing film dressing, a placebo version of the same (negative control) and the commercially available CHG-impregnated gel dressing (positive control). Serial dilutions of the surviving organisms were quantified using the pour plate after 1, 3, 5, and 7 days of incubation in order to calculate an antimicrobial log10 reduction for each organism/dressing combination at each point in time. Results The new CHG-containing film dressing delivered greater than 5.0 log10 reduction throughout the 7 days on all aerobic gram-negative bacilli and Staphylococcus species tested. As of day 1 the CHG-containing film dressing provided greater than 5.0 log10 reduction on Candida albicans. There were no statistically significant differences in the log10 reduction between the two dressings tested. Conclusion The new CHG-containing film dressing was found to be as effective as the chlorhexidine gluconate-impregnated gel dressing on clinically relevant microbes. PMID:26599087

Etiology of child mortality in Goroka, Papua New Guinea: a prospective two-year study.

PubMed Central

Duke, Trevor; Michael, Audrey; Mgone, Joyce; Frank, Dale; Wal, Tilda; Sehuko, Rebecca

2002-01-01

OBJECTIVE: To collect accurate data on disease- and microbial-specific causes and avoidable factors in child deaths in a developing country. METHODS: A systematic prospective audit of deaths of children seen at Goroka Hospital in the highlands of Papua New Guinea was carried out. Over a 24-month period, we studied 353 consecutive deaths of children: 126 neonates, 186 children aged 1-59 months, and 41 children aged 5-12 years. FINDINGS: The most frequent age-specific clinical diagnoses were as follows: for neonates--very low birth weight, septicaemia, birth asphyxia and congenital syphilis; for children aged 1-59 months--pneumonia, septicaemia, marasmus and meningitis; and for children aged 5-12 years--malignancies and septicaemia. At least one microbial cause of death was identified for 179 (50.7%) children and two or more were identified for 37 (10.5%). Nine microbial pathogens accounted for 41% of all childhood deaths and 76% of all deaths that had any infective component. Potentially avoidable factors were identified for 177 (50%) of deaths. The most frequently occurring factors were as follows: no antenatal care in high-risk pregnancies (8.8% of all deaths), very delayed presentation (7.9%), vaccine-preventable diseases (7.9%), informal adoption or child abandonment leading to severe malnutrition (5.7%), and lack of screening for maternal syphilis (5.4%). Sepsis due to enteric Gram-negative bacilli occurred in 87 (24.6%). The strongest associations with death from Gram- negative sepsis were adoption/abandonment leading to severe malnutrition, village births, and prolonged hospital stay. CONCLUSIONS: Reductions in child mortality will depend on addressing the commonest causes of death, which include disease states, microbial pathogens, adverse social circumstances and health service failures. Systematic mortality audits in selected regions where child mortality is high may be useful for setting priorities, estimating the potential benefit of specific and non-specific interventions, and providing continuous feedback on the quality of care provided and the outcome of health reforms. PMID:11884969

[Nosocomial infection in patients receiving a solid organ transplant or haematopoietic stem cell transplant].

PubMed

Moreno Camacho, Asunción; Ruiz Camps, Isabel

2014-01-01

Bacterial infections are the most common infections in solid organ transplant recipients. These infections occur mainly in the first month after transplantation and are hospital-acquired. Nosocomial infections cause significant morbidity and are the most common cause of mortality in this early period of transplantation. These infections are caused by multi-drug resistant (MDR) microorganisms, mainly Gram-negative enterobacteria, non-fermentative Gram-negative bacilli, enterococci, and staphylococci. The patients at risk of developing nosocomial bacterial infections are those previously colonized with MDR bacteria while on the transplant waiting list. Intravascular catheters, the urinary tract, the lungs, and surgical wounds are the most frequent sources of infection. Preventive measures are the same as those applied in non-immunocompromised, hospitalized patients except in patients at high risk for developing fungal infection. These patients need antifungal therapy during their hospitalization, and for preventing some bacterial infections in the early transplant period, patients need vaccinations on the waiting list according to the current recommendations. Although morbidity and mortality related to infectious diseases have decreased during the last few years in haematopoietic stem cell transplant recipients, they are still one of the most important complications in this population. Furthermore, as occurs in the general population, the incidence of nosocomial infections has increased during the different phases of transplantation. It is difficult to establish general preventive measures in these patients, as there are many risk factors conditioning these infections. Firstly, they undergo multiple antibiotic treatments and interventions; secondly, there is a wide variability in the degree of neutropenia and immunosuppression among patients, and finally they combine hospital and home stay during the transplant process. However, some simple measures could be implemented to improve the current situation. Copyright © 2014 Elsevier España, S.L. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

CHANGES IN THE MICROFLORA OF HADDOCK FILLETS AND SHUCKED SOFT-SHELLED CLAMS AFTER IRRADIATION WITH Co$sup 60$ GAMMA RAYS AND STORAGE TO 0 C AND 6 C

DOE Office of Scientific and Technical Information (OSTI.GOV)

Masurovsky, E.B.; Voss, J.S.; Goldblith, S.A.

1963-05-01

Microbiologic analyses were made on haddock fillets and shucked soft- shelled clams before and after treatment with doses of from 50000 to 800000 rad of Co/sup 60/ gamma rays. A decided change occurred in the constitution of the microbial populations of both products: from a mixed gram-negativegram-positive flora to a predominantly gram-positive flora immediately after irradiation. The majority of these surviving microorganisms were micrococci, spore-forming bacilli, yeasts, molds, and actinomyces. During storage at refrigeration temperatures above freezing, the microflora changed from the more radioresistant gram-positive species to the more prolific gram-negative psychrophilic species that flourish at low temperatures. Micrococci andmore » gram-positive rods declined somewhat during the rise of the actively proteolytic-lipolytic pseudomonads and related species, but still remained at high enough levels to contribute significantly to the spoilage observed at different times in storage. The eventual spoilage of haddock fillets was characterized by discoloration with water-soluble yellow, green, and red bacterial pigments; degradation of the tissue, by proteolytic and other microbial enzymes; and formation of volatile compounds that smelled putrid, rancid, and generally foul and pungent. Shucked soft-shelled clams displayed a different spoilage pattern, changing to a gelatinous mass with stale, acrid, putrid, sulfurous odor. These differences are attributable to the varied biochemical nature of the tissues involved, the Eh potential within the tissues, the time sequence of microfloral change, and the species and types of microorganisms associated with each product. Staphylococci were present in small numbers, but did not appear to be species of public health significance. Gram-negative enteric rods were also encountered, but were considered to be of doubtful public health importance. (BBB)« less

Hyperammoniemic coma in a patient with ureterosigmoidostomy and normal liver function.

PubMed

Van Laethem, J L; Gay, F; Franck, N; Van Gossum, A

1992-11-01

Hyperammoniemic encephalopathy has been reported after ureterosigmoidostomy. Its development is related to a problem of bacterial overgrowth and, most often, is favored by the presence of an underlying liver dysfunction. We report the case of a 43-year-old woman with a ureterosigmoidostomy done 28 years earlier who developed hyperammoniemic coma induced by an acute rectocolitis and in the absence of any detectable liver dysfunction. Neither administration of Lactilol and neomycin nor rectal tube drainage were effective; systemic antimicrobial therapy effective against the urease-producing gram-negative bacilli was required and led to a decrease in serum ammonia levels and a dramatic clinical improvement.

Septicemia in a neonatal calf associated with Chromobacterium violaceum.

PubMed

Ajithdoss, D K; Porter, B F; Calise, D V; Libal, M C; Edwards, J F

2009-01-01

Chromobacterium violaceum infections are highly fatal and rarely reported in domestic animals. This report describes a fatal case of C. violaceum septicemia in a 7-day-old female beef calf. The calf had necrosuppurative omphalophlebitis, necrotizing interstitial pneumonia, necrosuppurative hepatitis and splenitis, anterior uveitis with hypopyon, suppurative polyarthritis, and disseminated hemorrhagic meningitis with multifocal necrotizing encephalitis. Histologically, clusters of gram-negative bacilli were found in many of the lesions. C. violaceum was isolated in high numbers from the lungs, liver, spleen, carpus, and in pure culture from the cerebrospinal fluid. To our knowledge, this is the first case report of chromobacteriosis in a calf.

Protective efficacy of heat-inactivated B. thailandensis, B. mallei or B. pseudomallei against experimental melioidosis and glanders.

PubMed

Sarkar-Tyson, Mitali; Smither, Sophie J; Harding, S V; Atkins, Timothy P; Titball, Richard W

2009-07-16

Burkholderia pseudomallei and Burkholderia mallei are gram-negative bacilli that are the causative agents of melioidosis and glanders, respectively. Both humans and animals are susceptible to both diseases. There is currently no vaccine available for the prevention of disease. We report the protective efficacy of heat-inactivated Burkholderia thailandensis, B. mallei or B. pseudomallei cells as vaccines against murine melioidosis and glanders. Immunisation with heat-inactivated B. pseudomallei cells provided the highest levels of protection against either melioidosis or glanders. These studies indicate the longer term potential for heat-inactivated bacteria to be developed as vaccines against melioidosis and glanders.

Gram-negative diabetic foot osteomyelitis: risk factors and clinical presentation.

PubMed

Aragón-Sánchez, Javier; Lipsky, Benjamin A; Lázaro-Martínez, Jose L

2013-03-01

Osteomyelitis frequently complicates infections in the feet of patients with diabetes. Gram-positive cocci, especially Staphylococcus aureus, are the most commonly isolated pathogens, but gram-negative bacteria also cause some cases of diabetic foot osteomyelitis (DFO). These gram-negatives require different antibiotic regimens than those commonly directed at gram-positives. There are, however, few data on factors related to their presence and how they influence the clinical picture. We conducted a retrospective study to determine the variables associated with the isolation of gram-negative bacteria from bone samples in cases of DFO and the clinical presentation of these infections. Among 341 cases of DFO, 150 had a gram-negative isolate (alone or combined with a gram-positive isolate) comprising 44.0% of all patients and 50.8% of those with a positive bone culture. Compared with gram-positive infections, wounds with gram-negative organisms more often had a fetid odor, necrotic tissue, signs of soft tissue infection accompanying osteomyelitis, and clinically severe infection. By multivariate analysis, the predictive variables related to an increased likelihood of isolating gram-negatives from bone samples were glycated hemoglobin <7% (odds ratio [OR] = 2.0, 95% confidence interval [CI] = 1.1-3.5) and a wound caused by traumatic injury (OR = 2.0, 95% CI = 1.0-3.9). Overall, patients whose bone samples contained gram-negatives had a statistically significantly higher prevalence of leukocytosis and higher white blood cell counts than those without gram-negatives. In conclusion, gram-negative organisms were isolated in nearly half of our cases of DFO and were associated with more severe infections, higher white blood cell counts, lower glycated hemoglobin levels, and wounds of traumatic etiology.

Septic abortion presenting as a right lower trapezius abscess secondary to Bacteroides fragilis bacteraemia.

PubMed

Yamanaka, Yusuke; Shimabukuro, Akira

2017-08-21

A 43-year-old Japanese woman was evaluated in the outpatient department for right shoulder pain and fever, which began 5 days earlier. MRI of the right shoulder revealed a high-intensity area deep in the right trapezius muscle. Aspiration revealed purulent fluid, and Gram staining of the fluid showed Gram-negative bacilli. The patient was also found to be profoundly anaemic and to have a positive urine pregnancy test. On admission, we initiated intravenous ampicillin-sulbactam and aztreonam. She underwent dilatation and curettage for septic abortion and surgical drainage of the right shoulder abscess. Bacteroides fragilis was isolated from the blood, uterine aspiration and abscess samples. On hospital day 4, a whole-body CT scan revealed no other abscesses, and ampicillin-sulbactam was continued for 28 days. The patient was discharged on hospital day 29. Gram staining is an important tool for evaluating infectious aetiologies. © BMJ Publishing Group Ltd (unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Isolation and speciation of Prevotella strains from periodontal abscesses.

PubMed

Dumitriu, S; Băncescu, G; Murea, A; Skaug, N

1998-01-01

The aims of the study were to isolate and to identify at species level the Prevotella strains in pus samples collected by needle aspiration from 25 Romanian patients with periodontal abscesses. Gram-stained smears and cultures on selective and nonselective media were performed from each of the 25 pus samples. The isolates were identified on the basis of Gram staining, cultural characteristics and standard biochemical reactions. The Gram-negative anaerobic bacilli isolates were biochemically characterized and identified at species level using the Rapid ID 32 A system (Bio Mérieux, France). Fifteen Prevotella isolates belonging to one of the following species: P. melaninogenica, P. denticola, P. oralis, P. loescheii and P. bivia were recovered. All Prevotella isolates reacted similarly in 20 tests in the Rapid ID 32 A system. The P. melaninogenica strain showed approximately the same biochemical profile and only two sugar fermentation tests were not constantly positive. The study confirmed that Prevotella is often involved in periodontal abscesses (> 50% of the cases) in association with other anaerobic or/and aerobic bacteria. P. melaninogenica was the most frequently isolated Prevotella species from the investigated cases.

Soil microbial community responses to acid exposure and neutralization treatment.

PubMed

Shin, Doyun; Lee, Yunho; Park, Jeonghyun; Moon, Hee Sun; Hyun, Sung Pil

2017-12-15

Changes in microbial community induced by acid shock were studied in the context of potential release of acids to the environment due to chemical accidents. The responses of microbial communities in three different soils to the exposure to sulfuric or hydrofluoric acid and to the subsequent neutralization treatment were investigated as functions of acid concentration and exposure time by using 16S-rRNA gene based pyrosequencing and DGGE (Denaturing Gradient Gel Electrophoresis). Measurements of soil pH and dissolved ion concentrations revealed that the added acids were neutralized to different degrees, depending on the mineral composition and soil texture. Hydrofluoric acid was more effectively neutralized by the soils, compared with sulfuric acid at the same normality. Gram-negative ß-Proteobacteria were shown to be the most acid-sensitive bacterial strains, while spore-forming Gram-positive Bacilli were the most acid-tolerant. The results of this study suggest that the Gram-positive to Gram-negative bacterial ratio may serve as an effective bio-indicator in assessing the impact of the acid shock on the microbial community. Neutralization treatments helped recover the ratio closer to their original values. The findings of this study show that microbial community changes as well as geochemical changes such as pH and dissolved ion concentrations need to be considered in estimating the impact of an acid spill, in selecting an optimal remediation strategy, and in deciding when to end remedial actions at the acid spill impacted site. Copyright © 2017 Elsevier Ltd. All rights reserved.

Curative Treatment of Severe Gram-Negative Bacterial Infections by a New Class of Antibiotics Targeting LpxC

PubMed Central

Lemaître, Nadine; Liang, Xiaofei; Najeeb, Javaria; Lee, Chul-Jin; Titecat, Marie; Leteurtre, Emmanuelle; Simonet, Michel; Toone, Eric J.

2017-01-01

ABSTRACT The infectious diseases caused by multidrug-resistant bacteria pose serious threats to humankind. It has been suggested that an antibiotic targeting LpxC of the lipid A biosynthetic pathway in Gram-negative bacteria is a promising strategy for curing Gram-negative bacterial infections. However, experimental proof of this concept is lacking. Here, we describe our discovery and characterization of a biphenylacetylene-based inhibitor of LpxC, an essential enzyme in the biosynthesis of the lipid A component of the outer membrane of Gram-negative bacteria. The compound LPC-069 has no known adverse effects in mice and is effective in vitro against a broad panel of Gram-negative clinical isolates, including several multiresistant and extremely drug-resistant strains involved in nosocomial infections. Furthermore, LPC-069 is curative in a murine model of one of the most severe human diseases, bubonic plague, which is caused by the Gram-negative bacterium Yersinia pestis. Our results demonstrate the safety and efficacy of LpxC inhibitors as a new class of antibiotic against fatal infections caused by extremely virulent pathogens. The present findings also highlight the potential of LpxC inhibitors for clinical development as therapeutics for infections caused by multidrug-resistant bacteria. PMID:28743813

Comparative Evaluation of Four Phenotypic Tests for Detection of Carbapenemase-Producing Gram-Negative Bacteria

PubMed Central

Noël, Audrey; Berhin, Catherine; Hoebeke, Martin; Bouchahrouf, Warda; Yunus, Sami; Bogaerts, Pierre; Glupczynski, Youri

2016-01-01

ABSTRACT Four screening assays aimed for rapid detection of carbapenemase production from Gram-negative bacterial isolates, i.e., the Neo-Rapid Carb kit (Rosco Diagnostica A/S), the Rapidec Carba NP test (bioMérieux SA), the β Carba test (Bio-Rad Laboratories N.V.), and a homemade electrochemical assay (BYG Carba test) were evaluated against a panel comprising 328 clinical isolates (Enterobacteriaceae [n = 198] and nonfermentative Gram-negative bacilli [n = 130]) with previously characterized resistance mechanisms to carbapenems. Among Enterobacteriaceae isolates, the BYG Carba test and the β Carba test showed excellent sensitivities (respectively, 100% and 97.3%) and specificities (respectively, 98.9% and 97.7%). The two other assays yielded poorer performances with sensitivity and specificity of 91.9% and 83.9% for the Rapidec Carba NP test and of 89.2% and 89.7% for the Neo-Rapid Carb kit, respectively. Among Pseudomonas spp., sensitivities and specificities ranged, respectively, from 87.3% to 92.7% and from 88.2% to 94.1%. Finally, all tests performed poorly against Acinetobacter spp., with sensitivities and specificities, respectively, ranging from 27.3% to 75.8% and from 75 to 100%. Among commercially available assays, the β Carba test appeared to be the most convenient for routine use and showed the best overall performances, especially against OXA-48-like producers. The excellent performance of the BYG Carba test against Enterobacteriaceae was confirmed (100% sensitivity and 98.9% specificity). PMID:27927915

Usability application of multiplex polymerase chain reaction in the diagnosis of microorganisms isolated from urine of patients treated in cancer hospital

PubMed Central

Cybulski, Zefiryn; Schmidt, Katarzyna; Grabiec, Alicja; Talaga, Zofia; Bociąg, Piotr; Wojciechowicz, Jacek; Roszak, Andrzej; Kycler, Witold

2013-01-01

Background The objective of this study was: i) to compare the results of urine culture with polymerase chain reaction (PCR) -based detection of microorganisms using two commercially available kits, ii) to assess antimicrobial susceptibility of urine isolates from cancer patients to chosen antimicrobial drugs and, if necessary, to update the recommendation of empirical therapy. Materials and methods. A one-year hospital-based prospective study has been conducted in Greater Poland Cancer Centre and Genetic Medicine Laboratory CBDNA Research Centre in 2011. Urine cultures and urine PCR assay from 72 patients were examined Results Urine cultures and urine PCR assay from 72 patients were examined. Urine samples were positive for 128 strains from which 95 (74%) were identical in both tests. The most frequently isolated bacteria in both culture and PCR assay were coliform organisms and Enterococcus spp. The Gram negative bacilli were most resistant to cotrimoxazol. 77.2% of these bacilli and 100% of E. faecalis and S. agalactiae were sensitive to amoxicillin-clavulanic acid. 4.7% of Gram positive cocci were resistant to nitrofurantoin. Conclusions The PCR method quickly finds the causative agent of urinary tract infection (UTI) and, therefore, it can help with making the choice of the proper antimicrobial therapy at an early stage. It appears to be a viable alternative to the recommendations made in general treatment guidelines, in cases where diversified sensitivity patterns of microorganisms have been found. PMID:24133395

A prospective study on evaluation of pathogenesis, biofilm formation, antibiotic susceptibility of microbial community in urinary catheter

NASA Astrophysics Data System (ADS)

Younis, Khansa Mohammed; Usup, Gires; Ahmad, Asmat

2015-09-01

This study is aimed to isolate, detect biofilm formation ability and antibiotic susceptibility of urinary catheter adherent microorganisms from elderly hospitalized patient at the Universiti Kebangsaan Malaysia Medical Center. Microorganisms were isolated from three samples of urinary catheters (UC) surface; one of the acute vascular rejection patient (UCB) and two from benign prostate hyperplasia patients (UCC and UCD). A total of 100 isolates was isolated with 35 from UCB, 38 (UCC) and 28 (UCD). Ninety six were identified as Gram-negative bacilli, one Gram-positive bacilli and three yeasts. Results of biofilm forming on sterile foley catheter showed that all the isolates can form biofilm at different degrees; strong biofilm forming: 32% from the 35 isolates (UCB), 25% out of 38 isolates (UCC), 26% out of 28 isolates (UCD). As for moderate biofilm forming; 3% from UCB, 10% from UCC and 2% from UCD. Weak biofilm forming in UCC (3%). The antibiotic susceptibility for (UCB) isolates showed highly resistant to ampicillin, novobiocin and penicillin 100 (%), kanamycin (97%), tetracycline (94%), chloramphenicol (91%), streptomycin (77%) and showed low level of resistance to gentamycin (17%), while all the isolates from (UCC-D) showed high resistant towards ampicillin and penicillin, novobiocin (94%), tetracycline (61%), streptomycin (53%), gentamycin (50%) and low level of resistance to kanamycin (48%), chloramphenicol (47%). The findings indicate that these isolates can spread within the community on urinary catheters surface and produce strong biofilm, therefore, monitoring antibiotic susceptibility of bacteria isolated in the aggregation is recommended.

Multi-Drug–Resistant Gram-Negative Infections in Deployment-Related Trauma Patients

PubMed Central

Li, Ping; Whitman, Timothy J.; Blyth, Dana M.; Schnaubelt, Elizabeth R.; Mende, Katrin; Tribble, David R.

2017-01-01

Abstract Background: The contribution of multi-drug–resistant gram-negative bacilli infections (MDRGN-I) in patients with trauma is not well described. We present characteristics of MDRGN-Is among military personnel with deployment-related trauma (2009–2014). Patients and Methods: Data from the Trauma Infectious Disease Outcomes Study were assessed for infectious outcomes and microbial recovery. Infections were classified using standardized definitions. Gram-negative bacilli were defined as multi-drug–resistant if they showed resistance to ≥3 antibiotic classes or were producers of extended-spectrum β-lactamase or carbapenemases. Results: Among 2,699 patients admitted to participating U.S. hospitals, 913 (33.8%) experienced ≥1 infection event, of which 245 (26.8%) had a MDRGN-I. There were 543 MDRGN-I events (24.6% of unique 2,210 infections) with Escherichia coli (48.3%), Acinetobacter spp. (38.6%), and Klebsiella pneumoniae (8.4%) as the most common MDRGN isolates. Incidence of MDRGN-I was 9.1% (95% confidence interval [CI]: 8.0–10.2). Median time to MDRGN-I event was seven days with 75% occurring within 13 days post-trauma. Patients with MDRGN-Is had a greater proportion of blast injuries (84.1% vs. 62.5%; p < 0.0001), traumatic amputations (57.5% vs. 16.3%; p < 0.0001), and higher injury severity (82.0% had injury severity score ≥25 vs. 33.7%; p < 0.0001) compared with patients with either no infections or non-MDRGN-Is. Furthermore, MDRGN-I patients were more frequently admitted to the intensive care unit (90.5% vs. 48.5%; p < 0.0001), colonized with a MDRGN before infection (58.0% vs. 14.7%; p < 0.0001), and required mechanical ventilation (78.0% vs. 28.8% p < 0.0001). Antibiotic exposure before the MDRGN-I event was significantly higher across antibiotic classes except first generation cephalosporins and tetracyclines, which were very commonly used with all patients. Regarding outcomes, patients with MDRGN-Is had a longer length of hospitalization than the comparator group (53 vs. 18 days; p < 0.0001). Conclusions: We found a high rate of MDRGN-I in our population characterized by longer hospitalization and greater injury severity. These findings inform treatment and infection control decisions in the trauma patient population. PMID:29173084

[Analysis of characteristics of bacteria in respiratory tract infection in 2013-2016 in Heibei 3A hospital: a single-center report of 7 497 patients].

PubMed

Hou, Lili; Liu, Lili; Dang, Ping; Kang, Guannan; Zhang, Qinfeng; Li, Dongling

2017-09-01

To analyze the changes and characteristics of respiratory tract bacteria in Hebei 3A Hospital, and to provide new rationale for clinical diagnosis and treatment. A single-center retrospective analysis was conducted. 7 497 patients with respiratory tract infection admitted to Hebei Chest Hospital from January 2013 to December 2016 were enrolled. Deep sputum was collected, and the bacterial cultures and susceptibility analysis was conducted in sputum and upper respiratory secretions were collected by fiberoptic bronchoscopy. A total of 7 497 patients with respiratory tract infection were enrolled in the study, and 11 909 strains of 13 kinds of dominant pathogens were isolated. The dominant pathogens for respiratory tract infection were Monilia albican (23.7%), Klebsiella pneumoniae (12.9%), Pseudomonas aeruginosa (11.6%), Escherichia coli (9.5%), Candida glabrata (9.1%), Acinetobacter baumanii (7.9%), Aspergillus (6.7%), Stenotrophomonas maltophilia (4.5%), coagulase negative Staphylococcus (3.7%) and some species of Pseudomonas (3.7%), Staphylococcus aureus (3.0%), Aerobacter cloacae (1.9%), and Candida tropicalis (1.8%). A total of 6 198 strains of 7 kinds of Gram negative (G - ) bacilli infection dominant pathogens accounts for 52.0% of all infections, Klebsiella pneumonia (24.8%), Pseudomonas aeruginosa (22.3%), Escherichia coli (18.2%) and Acinetobacter baumanii (15.3%) were the main pathogens, and increased year by year. Susceptibility analysis showed that the preferred antibiotics for G - bacteria were carbapenems, followed by risperidone, sulbactam, cefepime, amikacin, and the third generation of cephalosporins. A total of 798 strains of 2 kinds of Gram positive (G + ) bacilli infection dominant pathogens accounted for 6.7% of all infections, were coagulase negative Staphylococcus (54.8%) and Staphylococcus aureus (45.2%), each had changed little by year. Susceptibility analysis showed that G + bacteria were sensitive to glycopeptides, followed by cefoxitin, cotrimoxazole, the tetracyclines, quinolones, azithromycin, erythromycin and so on. The advantages of 4 species of fungi were 4 913 strains, accounted for all of the 41.3% strains, with 57.5% of Candida albicans, and the trend was increasing year by year. Susceptibility analysis results showed that the antifungal susceptibility of dominant fungi were higher. G - bacilli is still the main source of infection, and showed an upward trend year by year. Fungal infection rate cannot be ignored, and we must pay attention to fungal infection incentives. We should strengthen the rational use of antibiotics.

Bacterial flora of combat wounds from eastern Ukraine and time-specified changes of bacterial recovery during treatment in Ukrainian military hospital.

PubMed

Valentine, Kovalchuk P; Viacheslav, Kondratiuk M

2017-04-07

Microbiology of modern war wounds is unique for each military conflict. Climatic and geographical features of the theater of war, contemporary warfare as well as wound management affect the microbial flora of wounds. This study was designed to determine time-specific microbial flora of combat wounds of upper and lower extremities obtained during the war in eastern Ukraine. The patients enrolled in study had combat wounds of upper or lower extremities which were treated in the Military Medical Clinical Center of Central Region. The wounds were swab-cultured and measured at each surgical debridement. The recovered microorganisms were identified and their antimicrobial resistance profiles were evaluated by disc diffusion method. Forty-nine patients with battle-field wounds were enrolled in the study from July to November 2014; all patients were male with a mean Injury Severity Score and arrival APACHE II scores of 16.2 ± 10.7 and 7.4 ± 4.2 respectively. Among 128 swab cultures, 100 swab cultures were positive. Swab cultures were obtained from 57 wounds of 49 patients. The results of the test showed that 87.7% of all positive swab cultures contained a single-organism while the rest of the swab-culture results showed polymicrobial growth. Among the isolated microorganisms 65% (76 strains) were Gram-negative rods, 22.2% (26 strains) of Gram-positive cocci, followed by Gram-positive rods (12.8%, 15 strains). We found that epidemiology of wound infection changes with the time after injury. The most common bacterial isolates cultured during the first week were Gram-positive microbes with low pathogenicity. The number of Gram-negative rods increased during the wound healing process. The incidence of Gram-positive microorganisms' growth fell after the first week and increased after third week. During wound healing, bacterial microflora of wounds changes with increasing number of Gram-negative rods with predominance of Acinetobacter species. Predominant microorganisms in positive swab-cultures after first week were nonfermentative Gram-negative bacilli (68% of swab-cultures), which in 53% of the swab-cultures belonged to the genus Acinetobacter, and in 15% to the genus Pseudomonas. The incidence of polymicrobial wound cultures increased from first week to second post-injury week. The most frequent microbial mixture were Acinetobacter baumannii with Enterobacteriaceae or other nonfermentative Gram negative rods with Enterococcus spp. We observed bacteria recovery from wounds during proliferation phase. These wounds had no pure inflammation signs and were free of devitalized tissues. Any wound is at some risk of becoming infected. In the event of infection, a wound fails to heal, treatment costs rise, and general wound management practices become more resource demanding. Determining the microorganisms which colonize battle wounds and cause wound infection is paramount. This information can help to treat battle wound infections or even changes infection control strategies. The fact of shifting in wound microbiology in the favor of bacteria responsible for healthcare-associated infections support to the proposition that these changes are nosocomially related [4, 14]. For Ukrainian military medicine this study is the first time-specified assessment of battle wound colonization from the World War II.

Symptomatic peripheral mycotic aneurysms due to infective endocarditis: a contemporary profile.

PubMed

González, Isabel; Sarriá, Cristina; López, Javier; Vilacosta, Isidre; San Román, Alberto; Olmos, Carmen; Sáez, Carmen; Revilla, Ana; Hernández, Miguel; Caniego, Jose Luis; Fernández, Cristina

2014-01-01

Peripheral mycotic aneurysms (PMAs) are a relatively rare but serious complication of infective endocarditis (IE). We conducted the current study to describe and compare the current epidemiologic, microbiologic, clinical, diagnostic, therapeutic, and prognostic characteristics of patients with symptomatic PMAs (SPMAs). A descriptive, comparative, retrospective observational study was performed in 3 tertiary hospitals, which are reference centers for cardiac surgery. From 922 definite IE episodes collected from 1996 to 2011, 18 patients (1.9%) had SPMAs. Because all SPMAs developed in left-sided IE, we performed a comparative study between 719 episodes of left-sided IE without SPMAs and 18 episodes with SPMAs. We found a higher frequency of intravenous drug abuse, native valve IE, intracranial bleeding, septic emboli, multiple embolisms, and IE diagnostic delay >30 days in patients with SPMAs than in patients without SPMAs. The causal microorganisms were gram-positive cocci (n =10), gram-negative bacilli (n = 2), gram-positive bacilli (n = 3), Bartonella henselae (n = 1), Candida albicans (n = 1), and negative culture (n = 1). The median IE diagnosis delay was 15 days (interquartile range [IQR], 13-33 d) in the case of high-virulence microorganisms versus 45 days (IQR, 30-240 d) in the case of low- to medium-virulence microorganisms. Twelve SPMAs were intracranial and 6 were extracranial. In 10 cases (8 intracranial and 2 extracranial), SPMAs were the initial presentation of IE; the remaining cases developed symptoms during or after finishing parenteral antibiotic treatment. The initial diagnosis of intracranial SPMAs was made by computed tomography (CT) or magnetic resonance imaging in 6 unruptured aneurysms and by angiography in 6 ruptured aneurysms. The initial test in extracranial SPMAs was Doppler ultrasonography in limbs, CT in liver, and coronary angiography in heart. Four (3 intracranial, 1 extracranial) of 7 (6 intracranial, 1 extracranial) patients treated only with antibiotics died. Surgical resection was performed in 7 (3 intracranial, 4 extracranial) and endovascular repair in 4 (3 intracranial, 1 extracranial) patients; all of them survived. In conclusion, we found that SPMAs were a rare complication of IE that developed only in left-sided IE, and especially in native valves. Intracranial hemorrhage, embolism, multiple embolisms, and diagnostic delay of IE were more common in patients with SPMAs. The microbiologic profile was diverse, but microorganisms of low-medium virulence were predominant, and had a greater delayed diagnosis of IE than those caused by microorganisms of high virulence. SPMAs were often the initial presentation of IE. The most common location of SPMAs was intracranial. Noninvasive radiologic imaging techniques were the initial imaging test in intracranial unruptured SPMAs and in most extracranial SPMAs. Surgical and endovascular treatments were safe and effective. Endovascular treatment could be the first line of treatment in selected cases. Mortality was high in those cases treated only with antibiotics.

Childhood pyogenic meningitis: clinical and investigative indicators of etiology and outcome.

PubMed Central

Johnson, Abdul-Wahab B. R.; Adedoyin, Olanrewaju T.; Abdul-Karim, Aishat A.; Olanrewaju, Abdul-Waheed I.

2007-01-01

The relevant parameters of 71 consecutive pediatric admissions for pyogenic meningitis at the University of Ilorin Teaching Hospital, Ilorin, Nigeria, were analyzed to identify possible clinical and nonmicrobiologic investigative clues of disease etiology and mortality. Cerebrospinal fluid (CSF) was Gram-smear positive (GSP) in 41 (57.6%) of the 71 cases. Twenty-three (56.1%) had Gram-positive cocci (GPC), 14 (34.2%) Gram-negative bacilli (GNB) and three (7.3%) Gram-negative diplococci (GND). The respective mean ages of GPC, GNB and GND cases were 4.49 +/- 5.3, 3.06 +/- 4.8 and 4.47 +/-4.9 years. Streptococcus pneumoniae accounted for 22 (78.6%) of the 28 CSF isolates (p=0.00), Haemophilus influenzae for two (7.1%) cases and Neisseria meningitides in one (3.5%). Anemia was significantly more common among GSP cases (p=0.04), as was convulsion among those with GNB-positive smears (p=0.03) and a bulging fontanelle in the Gram-smear-negative category. Otherwise, the prevalence and resolution times of the other clinical parameters were comparable across the etiological categories. There were 30 deaths (42.3%) among which GNB-positive cases had significantly shorter stay (p=0.045). Mortality was significantly higher in those with an abnormal respiratory rhythm at admission (p=0.04), purulent/turbid CSF (p=0.03), CSF protein of >150 mg/dl (p=0.02) and glucose <1 mg/dl (p=0.047). Our findings highlight the inherent limitations of predicting the etiology of pediatric meningitides from the clinical parameters as well as the poor prognostic import of respiratory dysrhythmia and a profoundly deranged CSF protein and glucose. The etiological burden of GPC/S. pneumoniae in childhood meningitides in sub-Saharan Africa, the propensity of GNB/H. influenzae for quick fatality and the need for the relevant preventive vaccines are expounded in the discussion. PMID:17722674

Prevotella maculosa sp. nov., isolated from the human oral cavity.

PubMed

Downes, Julia; Sutcliffe, Iain C; Booth, Veronica; Wade, William G

2007-12-01

Three strains of anaerobic Gram-negative bacilli isolated from human oral sites were subjected to a comprehensive range of phenotypic and genotypic tests and were found to comprise a homogeneous group. 16S rRNA gene sequence analysis revealed the strains to constitute a novel group within the genus Prevotella, most closely related to Prevotella oris and Prevotella salivae. A novel species, Prevotella maculosa sp. nov., is proposed to accommodate these strains. Prevotella maculosa is saccharolytic and produces acetic and succinic acids as end products of fermentation. The G+C content of the DNA of the type strain is 48 mol%. The type strain of Prevotella maculosa is W1609(T) (=DSM 19339(T) =CCUG 54766(T)).

[Study of bacterial flora in the oral cavity and stomach of elderly patients receiving nasogastric tube feeding].

PubMed

Matsuura, T; Suzuki, K; Yamakoshi, M; Yamamoto, T; Yamamoto, T; Yoshitomo, K; Tonegawa, K; Ariga, K; Odawara, F

1997-05-01

To investigate the significance of oropharyngeal flora and gastric flora in elderly patients receiving nasogastric tube feeding, throat secretions and gastric aspirates were cultured and the pH of the latter was measured. Of 116 bacterial isolates from throat secretions of 27 elderly patients, 30 were beta-streptococci and 28 were Pseudomonas aeruginosa. Bacteria isolated from gastric aspirates numbered 86 and 24 (27.9%) of them were the same species as those found in the throat secretions. Patients with gastric pH were below 3.5 had significantly lower concentrations of gram-negative bacili in gastric aspirates. We also studied oropharyngeal flora in 33 elderly patients who were admitted to Nagoyashi Koseiin Geriatric Hospital. The major bacterial isolates from throat swabs of bedridden patients were gram-negative bacilli and beta-streptococci, especially group B streptococci (GBS). We measured the level of antibody to GBS in these patients. Those from whom GBS were isolated had high titers. These results suggest that in elderly patients receiving enteral nasogastric) tube feeding, large numbers of bacteria colonize the oral cavity and stomach. The measurement of type-specific antibody to GBS may be useful in managing such patients.

Rhodotorula glutinis meningitis: a case report and review of literature.

PubMed

Menon, Sarala; Gupta, H R; Sequeira, R; Chavan, Shazia; Gholape, D; Amandeep, S; Bhilave, N; Chowdhary, A S

2014-07-01

Rhodotorula is ubiquitous saprophytic yeast belonging to phylum Basidiomycota. These encapsulated basidiomycetes are being increasingly recognised as important emerging human pathogens. There are scanty reports of meningitis caused by Rhodurorula spp in HIV infected patients. We present one such case of meningitis by Rhodutorula glutinis in HIV-infected patient. The patient also had a past history of abdominal tuberculosis. The diagnosis of Rhodotorula was confirmed by Gram staining and culture of the cerebrospinal fluid (CSF). Contamination was ruled out by repeated culturing of CSF from the same patient. Therapy with Amphotericin B showed good results. Patient was discharged from the hospital. However, in the seventh month of follow-up patient was readmitted with complaints of fever, breathlessness, altered sensorium, vomiting and succumbed to his illness. This time the CSF cultures remained negative for Rhodotorula, acid fast bacilli and other pyogenic organisms. Our last 11-year retrospective analysis of 8197 specimens received for mycological work-up showed that this is the first report of R. glutinis isolation from our institute. © 2014 Blackwell Verlag GmbH.

Emerging drugs in sepsis.

PubMed

Leone, Marc; Textoris, Julien; Michel, Fabrice; Wiramus, Sandrine; Martin, Claude

2010-03-01

Sepsis remains a major cause of death in intensive care units. Despite an intense research, a new drug that is effective in reducing mortality in sepsis is still awaited. The literature was analyzed with Pubmed() during the 2008 - 2009 period. If required, seminal articles published before 2008 were cited. Clinical trials focusing on 'sepsis' were first assessed. Next, relevant experimental data in this field were reported. The goal of the review is to determine the role for new licensed antibiotics, to give an insight into the conflict on adjuvant therapies and to disclose new experimental concepts. New licensed antibiotics will offer the opportunity to refine the treatment choices. Direct hemoperfusion using polymyxin B-immobilized fiber column may be an option in sepsis due to Gram-negative bacilli. Among non-antibiotic drugs, new ongoing studies will clarify the role of drotrecogin alfa (activated) and low dose hydrocortisone. The modulation of monocytic human leukocyte antigen-DR seems the most prominent treatment. The use of cardiovascular drugs requires well-conducted clinical trials. The regulation of high mobility group box 1, adenosine blockade or correction of the impaired energy production is still at the experimental level.

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

Innate immune system capabilities of the Asian citrus psyllid, Diaphorina citri.

PubMed

Arp, Alex P; Martini, Xavier; Pelz-Stelinski, Kirsten S

2017-09-01

Citrus production worldwide is currently threatened by Huanglongbing, or citrus greening disease. The associated pathogen, Candidatus Liberibacter asiaticus (CLas), is transmitted by the Asian citrus psyllid, Diaphorina citri. Annotation of the D. citri genome revealed a reduced innate immune system lacking a number of antimicrobial peptides and the Imd pathway associated with defense against Gram-negative bacteria. We characterized this apparent immune reduction in survival assays in which D. citri were exposed to Gram-negative or Gram-positive bacteria. D. citri experienced significant mortality when exposed to Serratia marcescens (Gram-negative) through oral ingestion or by septic injury. Escherichia coli (Gram-negative) also caused significant D. citri mortality, but only when inoculated at high concentrations through oral ingestion or by septic injury. Neither Micrococcus luteus (Gram-positive) or Bacillus subtilis (Gram-positive) caused significant mortality as compared to controls in any experiment. E. coli titers increased rapidly following exposure, while M. luteus titer remained stable for 72 h. We demonstrate that D. citri is capable of defending against E. coli, a Gram-negative bacterium, despite lacking the Imd defense pathway. The tolerance of D. citri to M. luteus infection, yet inability to effectively clear infections, presents questions to efficacy of D. citri immune response to effectively clear Gram-positive infections. Copyright © 2017. Published by Elsevier Inc.

Monolateral suppurative parotitis in a neonate and review of literature.

PubMed

Decembrino, Lidia; Ruffinazzi, Giulia; Russo, Fabio; Manzoni, Paolo; Stronati, Mauro

2012-07-01

Neonatal suppurative parotitis is a rare condition characterized by swelling, pain, and erythema over the affected gland. Antimicrobials and adequate hydration are an essential part of treatment. Surgical intervention is reserved for organized abscesses and for infections not responding to medical management. A case report and review of the literature. Only few case reports and case series are reported in literature on neonatal parotitis. Transmission of bacteria seems to occur mainly by ascending spread through Stensen's duct, or by hematogenous spread from a distant focus. Dehydration, low birth weight, immune suppression, ductal obstruction, oral trauma and structural abnormalities of the parotid gland are recognised as risk factors. The most common pathogen is Staphylococcus aureus. Other less frequent agents are other Gram-positive cocci, Gram-negative bacilli and rarely anaerobic bacteria. Advances in antimicrobial therapy have improved both outcome and prognosis. Thanks to the prompt antibiotic treatment complications are now drastically reduced. Ultrasound examination may help in the diagnosis and monitoring of clinical course. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

[Prevotella species involved in the onset of superficial face and neck abscesses].

PubMed

Băncescu, Gabriela; Dumitriu, Silvia; Băncescu, A; Preoteasa, Elena; Skaug, N

2004-01-01

The aim of the study was to identify at species level the Prevotella strains from 30 pus samples collected by needle aspiration in patients with abscesses of fascial spaces of the face and neck. Microscopy of Gram-stained direct smear and cultures on selective and nonselective media incubated aerobically and anaerobically were performed in each pus specimen. All strains of Gram-negative strictly anaerobic bacilli were identified to genus and species level using conventional methods of diagnosis and the Rapid ID 32 A system (BioMérieux, Marcy-l'Etoile, France). The Prevotella isolates were recovered (alone or in association with other bacteria) in 46.6% of the investigated specimens and were represented in reverse order of frequency by the following species: P. melaninogenica, P. denticola, P. oralis and P. loescheii. In conclusion, the rate of Prevotella isolation was relatively high and the most frequently isolated species was P. melaninogenica (followed by P. denticola), while no P. intermedia was found.

[Placental transfer and pharmacokinetic parameters of flomoxef during the perinatal period].

PubMed

Makinoda, S; Tsuruta, H; Iwaki, M; Negishi, H; Hanatani, K; Tanaka, T; Fujimoto, S

1991-06-01

Flomoxef (FMOX), a new oxacephem with low MIC values against not only Gram-negative bacilli (GNB) but also against Gram-positive cocci (GPC), was evaluated for its transfer into fetus, amniotic fluid, maternal milk, spinal fluid and urine during the perinatal period following a single intravenous drip infusion at a dose of 1 g for 30 minutes. The results obtained are summarized below. 1. High concentrations of FMOX were demonstrated in maternal serum, umbilical arterial serum and amniotic fluid with Cmax values of 48.0, 10.99 and 10.20 micrograms/ml, respectively. 2. Maternal urinary excretion rate was 65.4% in the first 6 hours after administration. 3. In contrast, maternal milk and spinal fluid levels were lower than 3 and 0.20 micrograms/ml, respectively. These results showed a good placental transfer of FMOX, which is very useful for various perinatal infections. No adverse effects were observed in mothers and neonates during the course of this study.

Risk score for identifying adults with CSF pleocytosis and negative CSF Gram stain at low risk for an urgent treatable cause.

PubMed

Hasbun, Rodrigo; Bijlsma, Merijn; Brouwer, Matthijs C; Khoury, Nabil; Hadi, Christiane M; van der Ende, Arie; Wootton, Susan H; Salazar, Lucrecia; Hossain, Md Monir; Beilke, Mark; van de Beek, Diederik

2013-08-01

We aimed to derive and validate a risk score that identifies adults with cerebrospinal fluid (CSF) pleocytosis and a negative CSF Gram stain at low risk for an urgent treatable cause. Patients with CSF pleocytosis and a negative CSF Gram stain were stratified into a prospective derivation (n = 193) and a retrospective validation (n = 567) cohort. Clinically related baseline characteristics were grouped into three composite variables, each independently associated with a set of predefined urgent treatable causes. We subsequently derived a risk score classifying patients into low (0 composite variables present) or high (≥ 1 composite variables present) risk for an urgent treatable cause. The sensitivity of the risk score was determined in the validation cohort and in a prospective case series of 214 adults with CSF-culture proven bacterial meningitis, CSF pleocytosis and a negative Gram stain. A total of 41 of 193 patients (21%) in the derivation cohort and 71 of 567 (13%) in the validation cohort had an urgent treatable cause. Sensitivity of the dichotomized risk score to detect an urgent treatable cause was 100.0% (95% CI 93.9-100.0%) in the validation cohort and 100.0% (95% CI 97.8-100.0%) in bacterial meningitis patients. The risk score can be used to identify adults with CSF pleocytosis and a negative CSF Gram stain at low risk for an urgent treatable cause. Copyright © 2013 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

RISK SCORE FOR IDENTIFYING ADULTS WITH CSF PLEOCYTOSIS AND NEGATIVE CSF GRAM STAIN AT LOW RISK FOR AN URGENT TREATABLE CAUSE

PubMed Central

Hasbun, Rodrigo; Bijlsma, Merijn; Brouwer, Matthijs C; Khoury, Nabil; Hadi, Christiane M; van der Ende, Arie; Wootton, Susan H.; Salazar, Lucrecia; Hossain, Md Monir; Beilke, Mark; van de Beek, Diederik

2013-01-01

Background We aimed to derive and validate a risk score that identifies adults with cerebrospinal fluid (CSF) pleocytosis and a negative CSF Gram stain at low risk for an urgent treatable cause. Methods Patients with CSF pleocytosis and a negative CSF Gram stain were stratified into a prospective derivation (n=193) and a retrospective validation (n=567) cohort. Clinically related baseline characteristics were grouped into three composite variables, each independently associated with a set of predefined urgent treatable causes. We subsequently derived a risk score classifying patients into low (0 composite variables present) or high ( ≥ 1 composite variables present) risk for an urgent treatable cause. The sensitivity of the risk score was determined in the validation cohort and in a prospective case series of 214 adults with CSF-culture proven bacterial meningitis, CSF pleocytosis and a negative Gram stain. Findings A total of 41 of 193 patients (21%) in the derivation cohort and 71 of 567 (13%) in the validation cohort had an urgent treatable cause. Sensitivity of the dichotomized risk score to detect an urgent treatable cause was 100.0% (95%CI 93.9-100.0%) in the validation cohort and 100.0% (95%CI 97.8-100.0%) in bacterial meningitis patients. Interpretation The risk score can be used to identify adults with CSF pleocytosis and a negative CSF Gram stain at low risk for an urgent treatable cause. PMID:23619080

Detection of biological threats. A challenge for directed molecular evolution.

PubMed

Petrenko, Valery A; Sorokulova, Iryna B

2004-08-01

The probe technique originated from early attempts of Anton van Leeuwenhoek to contrast microorganisms under the microscope using plant juices, successful staining of tubercle bacilli with synthetic dyes by Paul Ehrlich and discovery of a stain for differentiation of gram-positive and gram-negative bacteria by Hans Christian Gram. The technique relies on the principle that pathogens have unique structural features, which can be recognized by specifically labeled organic molecules. A hundred years of extensive screening efforts led to discovery of a limited assortment of organic probes that are used for identification and differentiation of bacteria. A new challenge--continuous monitoring of biological threats--requires long lasting molecular probes capable of tight specific binding of pathogens in unfavorable conditions. To respond to the challenge, probe technology is being revolutionized by utilizing methods of combinatorial chemistry, phage display and directed molecular evolution. This review describes how molecular evolution methods are applied for development of peptide, antibody and phage probes, and summarizes the author's own data on development of landscape phage probes against Salmonella typhimurium. The performance of the probes in detection of Salmonella is illustrated by a precipitation test, enzyme-linked immunosorbent assay (ELISA), fluorescence-activated cell sorting (FACS) and fluorescent, optical and electron microscopy.

Selective decontamination of the digestive tract ameliorates severe burn-induced insulin resistance in rats.

PubMed

Li, Jun; Zhu, Liang; Xu, Ming; Han, Juntao; Bai, Xiaozhi; Yang, Xuekang; Zhu, Huayu; Xu, Jie; Zhang, Xing; Gong, Yangfan; Hu, Dahai; Gao, Feng

2015-08-01

Severe burns often initiate the prevalence of hyperglycemia and insulin resistance, significantly contributing to adverse clinical outcomes. However, there are limited treatment options. This study was designed to investigate the role and the underlying mechanisms of oral antibiotics to selectively decontaminate the digestive tract (SDD) on burn-induced insulin resistance. Rats were subjected to 40% of total body surface area full-thickness burn or sham operation with or without SDD treatment. Translocation of FITC-labeled LPS was measured at 4h after burn. Furthermore, the effect of SDD on post-burn quantity of gram-negative bacteria in gut was investigated. Serum or muscle LPS and proinflammatory cytokines were measured. Intraperitoneal glucose tolerance test and insulin tolerance test were used to determine the status of systemic insulin resistance. Furthermore, intracellular insulin signaling (IRS-1 and Akt) and proinflammatory related kinases (JNK and IKKβ) were assessed by western blot. Burn increased the translocation of LPS from gut 4h after injury. SDD treatment effectively inhibited post-burn overgrowth of gram-negative enteric bacilli in gut. In addition, severe burns caused significant increases in the LPS and proinflammatory cytokines levels, activation of proinflammatory related kinases, and systemic insulin resistance as well. But SDD treatment could significantly attenuate burn-induced insulin resistance and improve the whole-body responsiveness to insulin, which was associated with the inhibition of gut-derived LPS, cytokines, proinflammatory related kinases JNK and IKKβ, as well as activation of IRS-1 and Akt. SDD appeared to have an effect on proinflammatory signaling cascades and further reduced severe burn-induced insulin resistance. Copyright © 2015 Elsevier Ltd and ISBI. All rights reserved.

Risk Factors for Urinary Tract Infections in Renal Allograft Recipients: Experience of a Tertiary Care Center in Hyderabad, South India.

PubMed

Mohan, M V N L R; Neeraja, M; Sudhaharan, S; Raju, S B; Gangadhar, T; Lakshmi, V

2017-01-01

Renal transplantation is an effective and commonly performed procedure for end-stage renal disease. Urinary tract infections are a major cause of morbidity and mortality in renal transplant patients. As data on postrenal transplant urinary tract infections from the Indian subcontinent are limited, the present study was conducted to estimate the burden of urinary tract infections in this vulnerable group of patients. This was a prospective study on patients undergoing renal transplantation in 2014 at our tertiary hospital in South India with a follow-up of 2 years to evaluate the risk factors for urinary tract infections. The prevalence of urinary tract infections was 41.9% with a male preponderance of 76.9%. Mean age of the 31 patients was 32.4 ± 10.2 years (range: 16-55 years). Gram-negative bacilli were the most common isolates with Escherichia coli being the predominant pathogen (53.3%). All the infections occurred within 1 year of transplantation with delayed graft function ( P < 0.001; confidence interval [CI]: 29.0-96.3) and prolonged hospital stay ( P = 0.0281; CI: 42.1-99.6) being the significant risk factors for acquiring urinary tract infections. Carbapenemase production was noted in 33.3% of isolates and all the Gram-negative organisms isolated in the 1 st month of transplantation were carbapenem-resistant (CR) E. coli . The high rate of carbapenem-resistant organisms in the early posttransplant period is a point of concern, especially with cadaver transplants. Infection control practices and catheter care need to be strictly monitored to minimize the risk for UTI in the immediate posttransplant period.

Development of the Nasopharyngeal Microbiota in Infants with Cystic Fibrosis.

PubMed

Prevaes, Sabine M P J; de Winter-de Groot, Karin M; Janssens, Hettie M; de Steenhuijsen Piters, Wouter A A; Tramper-Stranders, Gerdien A; Wyllie, Anne L; Hasrat, Raiza; Tiddens, Harm A; van Westreenen, Mireille; van der Ent, Cornelis K; Sanders, Elisabeth A M; Bogaert, Debby

2016-03-01

Cystic fibrosis (CF) is characterized by early structural lung disease caused by pulmonary infections. The nasopharynx of infants is a major ecological reservoir of potential respiratory pathogens. To investigate the development of nasopharyngeal microbiota profiles in infants with CF compared with those of healthy control subjects during the first 6 months of life. We conducted a prospective cohort study, from the time of diagnosis onward, in which we collected questionnaires and 324 nasopharynx samples from 20 infants with CF and 45 age-matched healthy control subjects. Microbiota profiles were characterized by 16S ribosomal RNA-based sequencing. We observed significant differences in microbial community composition (P < 0.0002 by permutational multivariate analysis of variance) and development between groups. In infants with CF, early Staphylococcus aureus and, to a lesser extent, Corynebacterium spp. and Moraxella spp. dominance were followed by a switch to Streptococcus mitis predominance after 3 months of age. In control subjects, Moraxella spp. enrichment occurred throughout the first 6 months of life. In a multivariate analysis, S. aureus, S. mitis, Corynebacterium accolens, and bacilli were significantly more abundant in infants with CF, whereas Moraxella spp., Corynebacterium pseudodiphtericum and Corynebacterium propinquum and Haemophilus influenzae were significantly more abundant in control subjects, after correction for age, antibiotic use, and respiratory symptoms. Antibiotic use was independently associated with increased colonization of gram-negative bacteria such as Burkholderia spp. and members of the Enterobacteriaceae bacteria family and reduced colonization of potential beneficial commensals. From diagnosis onward, we observed distinct patterns of nasopharyngeal microbiota development in infants with CF under 6 months of age compared with control subjects and a marked effect of antibiotic therapy leading toward a gram-negative microbial composition.

Presence, distribution and molecular epidemiology of multi-drug-resistant Gram-negative bacilli from medical personnel of intensive care units in Tianjin, China, 2007-2015.

PubMed

Liu, H; Fei, C N; Zhang, Y; Liu, G W; Liu, J; Dong, J

2017-06-01

Multi-drug-resistant Gram-negative bacteria (MDRGNB) have become an important cause of nosocomial infection in intensive care units (ICUs). To investigate the molecular epidemiology of MDRGNB isolated from medical personnel (MP) and non-medical personnel (NMP) at 69 ICUs in Tianjin, China. From April 2007 to October 2015, 2636 nasal and hand swab samples from 1185 MP and 133 NMP were cultured for GNB (including MDRGNB), meticillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). The susceptibilities of GNB to 14 antimicrobial agents were determined, and 80 MDRGNB were characterized using pulsed-field gel electrophoresis (PFGE) and dendrogram analysis. In total, 301 GNB were identified in 269 MP, including 109 MDRGNB isolates in 104 MP. Forty-two GNB were isolated from 39 NMP, which included 20 NMP with MDRGNB. Overall, 8.8% of MP were colonized with MDRGNB, which greatly exceeded colonization rates with MRSA (0.9%) and VRE (0.1%). Three pairs of Klebsiella pneumoniae and one pair of Enterobacter aerogenes were indistinguishable from each other, but the majority of isolate tests had distinct PFGE profiles. The prevalence of MDRGNB was high among ICU MP in Tianjin, and greatly exceeded that of VRE and MRSA. There was no difference in the rates of nasal carriage of MDRGNB between MP and NMP, but NMP were significantly more likely to have hand colonization with MDRGNB. PFGE profiles showed that there was only limited sharing of strains of MDR E. aerogenes and K. pneumoniae between personnel. Copyright © 2017 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

Multidrug and co-resistance patterns of non-fermenting Gram-negative bacilli involved in ventilator-associated pneumonia carrying class 1 integron in the North of Iran.

PubMed

Bagheri-Nesami, Masoumeh; Rezai, Mohammad Sadegh; Ahangarkani, Fatemeh; Rafiei, Alireza; Nikkhah, Attieh; Eslami, Gohar; Shafahi, Kheironesa; Hajalibeig, Azin; Khajavi, Rezvan

2017-09-01

Ventilator-associated pneumonia (VAP) due to non-fermenting Gram-negative bacilli (NFGNB), especially Pseudomonas aeruginosa and Acinetobacter spp., is one of the main hospital-acquired infections leading to mortality and morbidity, especially in intensive care units (ICUs). This study seeks to determine the multidrug and co-resistance (MDR) patterns of NFGNB that are agents of VAP, and assess the presence of class 1 integron in these bacteria. This cross-sectional study involved VAP patients admitted in the ICUs of 18 hospitals in the Mazandaran province, located in the North of Iran. The antibiotic susceptibility pattern was determined by the minimum inhibitory concentration (MIC) test by using broth microdilution method. Presence of class 1 integron was evaluated by the polymerase chain reaction (PCR) assay. Out of a total of 83 patients who were microbiologically diagnosed as VAP, 52 non-duplicated NFGNBs (24 P. aeruginosa and 28 A. baumannii ) were causative of VAP, out of which MDR NFGNBs were responsible for 48 (57.83%) cases. The frequencies of MDR NFGNBs were as follows: 27 (56.25%) A. baumannii and 21 (43.75%) P. aeruginosa . P. aeruginosa isolates were resistant to all aminoglycoside antibiotics (50%), ciprofloxacin (45.8%), ceftazidime (70.8%), cefepime (87.5%), colistin (62.5%), and imipenem (29.2%). A. baumannii isolates were resistant to aminoglycosides (53.6%), ciprofloxacin (85.7%), ceftazidime (92. 9%), cefepime (92.9%), colistin (35.7%), and imipenem (57.1%). Twelve isolates were resistant to all 10 tested antibiotics. The number of rates of class 1 integron, positive for MDR P. aeruginosa and MDR A. baumannii , were 20 (95.23%) and 21 (77.78%), respectively. The high prevalence of multidrug resistance and incidence of class 1 integron is a therapeutic concern. Employing antibiotic stewardship in hospitals could prevent the dissemination of MDR bacteria.

Outcome of Acute Prosthetic Joint Infections Due to Gram-Negative Bacilli Treated with Open Debridement and Retention of the Prosthesis▿

PubMed Central

Martínez-Pastor, Juan C.; Muñoz-Mahamud, Ernesto; Vilchez, Félix; García-Ramiro, Sebastián; Bori, Guillem; Sierra, Josep; Martínez, José A.; Font, Lluis; Mensa, Josep; Soriano, Alex

2009-01-01

The aim of our study was to evaluate the outcome of acute prosthetic joint infections (PJIs) due to gram-negative bacilli (GNB) treated without implant removal. Patients with an acute PJI due to GNB diagnosed from 2000 to 2007 were prospectively registered. Demographics, comorbidity, type of implant, microbiology data, surgical treatment, antimicrobial therapy, and outcome were recorded. Classification and regression tree analysis, the Kaplan-Meier survival method, and the Cox regression model were applied. Forty-seven patients were included. The mean age was 70.7 years, and there were 15 hip prostheses and 32 knee prostheses. The median number of days from the time of arthroplasty was 20. The most frequent pathogens were members of the Enterobacteriaceae family in 41 cases and Pseudomonas spp. in 20 cases. Among the Enterobacteriaceae, 14 were resistant to ciprofloxacin, while all Pseudomonas aeruginosa isolates were susceptible to ciprofloxacin. The median durations of intravenous and oral antibiotic treatment were 14 and 64 days, respectively. A total of 35 (74.5%) patients were in remission after a median follow-up of 463 days (interquartile range, 344 to 704) days. By use of the Kaplan-Meier survival curve, a C-reactive protein (CRP) concentration of ≤15 mg/dl (P = 0.03) and receipt of a fluoroquinolone, when all GNB isolated were susceptible (P = 0.0009), were associated with a better outcome. By use of a Cox regression model, a CRP concentration of ≤15 mg/dl (odds ratio [OR], 3.57; 95% confidence interval [CI], 1.05 to 12.5; P = 0.043) and receipt of a fluoroquinolone (OR, 9.09; 95% CI, 1.96 to 50; P = 0.005) were independently associated with better outcomes. Open debridement without removal of the implant had a success rate of 74.5%, and the factors associated with good prognosis were a CRP concentration at the time of diagnosis ≤15 mg/dl and treatment with a fluoroquinolone. PMID:19687237

The Increasing Challenge of Multidrug-Resistant Gram-Negative Bacilli

PubMed Central

Giuffrè, Mario; Geraci, Daniela M.; Bonura, Celestino; Saporito, Laura; Graziano, Giorgio; Insinga, Vincenzo; Aleo, Aurora; Vecchio, Davide; Mammina, Caterina

2016-01-01

Abstract Colonization and infection by multidrug-resistant gram-negative bacilli (MDR GNB) in neonatal intensive care units (NICUs) are increasingly reported. We conducted a 5-year prospective cohort surveillance study in a tertiary NICU of the hospital “Paolo Giaccone,” Palermo, Italy. Our objectives were to describe incidence and trends of MDR GNB colonization and the characteristics of the most prevalent organisms and to identify the risk factors for colonization. Demographic, clinical, and microbiological data were prospectively collected. Active surveillance cultures (ASCs) were obtained weekly. Clusters of colonization by extended spectrum β-lactamase (ESBL) producing Escherichia coli and Klebsiella pneumoniae were analyzed by conventional and molecular epidemiological tools. During the study period, 1152 infants were enrolled in the study. Prevalences of colonization by MDR GNB, ESBL-producing GNB and multiple species/genera averaged, respectively, 28.8%, 11.7%, and 3.7%. Prevalence and incidence density of colonization by MDR GNB and ESBL-producing GNB showed an upward trend through the surveillance period. Rates of ESBL-producing E coli and K pneumoniae colonization showed wide fluctuations peaking over the last 2 years. The only independent variables associated with colonization by MDR GNB and ESBL-producing organisms and multiple colonization were, respectively, the days of NICU stay (odds ratio [OR] 1.041), the days of exposure to ampicillin–sulbactam (OR 1.040), and the days of formula feeding (OR 1.031). Most clusters of E coli and K pneumoniae colonization were associated with different lineages. Ten out of 12 clusters had an outborn infant as their index case. Our study confirms that MDR GNB are an increasing challenge to NICUs. The universal once-a-week approach allowed us to understand the epidemiology of MDR GNB, to timely detect new clones and institute contact precautions, and to assess risk factors. Collection of these data can be an important tool to optimize antimicrobials use and control the emergence and dissemination of resistances in NICU. PMID:26962817

Outcome of acute prosthetic joint infections due to gram-negative bacilli treated with open debridement and retention of the prosthesis.

PubMed

Martínez-Pastor, Juan C; Muñoz-Mahamud, Ernesto; Vilchez, Félix; García-Ramiro, Sebastián; Bori, Guillem; Sierra, Josep; Martínez, José A; Font, Lluis; Mensa, Josep; Soriano, Alex

2009-11-01

The aim of our study was to evaluate the outcome of acute prosthetic joint infections (PJIs) due to gram-negative bacilli (GNB) treated without implant removal. Patients with an acute PJI due to GNB diagnosed from 2000 to 2007 were prospectively registered. Demographics, comorbidity, type of implant, microbiology data, surgical treatment, antimicrobial therapy, and outcome were recorded. Classification and regression tree analysis, the Kaplan-Meier survival method, and the Cox regression model were applied. Forty-seven patients were included. The mean age was 70.7 years, and there were 15 hip prostheses and 32 knee prostheses. The median number of days from the time of arthroplasty was 20. The most frequent pathogens were members of the Enterobacteriaceae family in 41 cases and Pseudomonas spp. in 20 cases. Among the Enterobacteriaceae, 14 were resistant to ciprofloxacin, while all Pseudomonas aeruginosa isolates were susceptible to ciprofloxacin. The median durations of intravenous and oral antibiotic treatment were 14 and 64 days, respectively. A total of 35 (74.5%) patients were in remission after a median follow-up of 463 days (interquartile range, 344 to 704) days. By use of the Kaplan-Meier survival curve, a C-reactive protein (CRP) concentration of < or = 15 mg/dl (P = 0.03) and receipt of a fluoroquinolone, when all GNB isolated were susceptible (P = 0.0009), were associated with a better outcome. By use of a Cox regression model, a CRP concentration of < or = 15 mg/dl (odds ratio [OR], 3.57; 95% confidence interval [CI], 1.05 to 12.5; P = 0.043) and receipt of a fluoroquinolone (OR, 9.09; 95% CI, 1.96 to 50; P = 0.005) were independently associated with better outcomes. Open debridement without removal of the implant had a success rate of 74.5%, and the factors associated with good prognosis were a CRP concentration at the time of diagnosis < or = 15 mg/dl and treatment with a fluoroquinolone.

Impact of imipenem and amikacin pharmacokinetic/pharmacodynamic parameters on microbiological outcome of Gram-negative bacilli ventilator-associated pneumonia.

PubMed

Pajot, O; Burdet, C; Couffignal, C; Massias, L; Armand-Lefevre, L; Foucrier, A; Da Silva, D; Lasocki, S; Laouénan, C; Mentec, H; Mentré, F; Wolff, M

2015-05-01

Despite recent advances, antibiotic therapy of ventilator-associated pneumonia (VAP) in ICU patients is still challenging. We assessed the impact of imipenem and amikacin pharmacokinetic and pharmacodynamic parameters on microbiological outcome in these patients. Patients with Gram-negative bacilli (GNB) VAP were prospectively included. Blood samples for pharmacokinetic analysis were collected after empirical administration of a combination of imipenem three times daily and one single dose of amikacin. MICs were estimated for each GNB obtained from respiratory samples. Microbiological success was defined as a ≥10(3) cfu/mL decrease in bacterial count in quantitative cultures between baseline and the third day of treatment. Thirty-nine patients [median (min-max) age = 60 years (28-84) and median SAPS2 at inclusion = 40 (19-73)] were included. Median MICs of imipenem and amikacin were 0.25 mg/L (0.094-16) and 2 mg/L (1-32), respectively. Median times over MIC and over 5× MIC for imipenem were 100% (8-100) and 74% (3-100), respectively. The median C1/MIC ratio for amikacin was 23 (1-76); 34 patients (87%) achieved a C1/MIC ≥10. Microbiological success occurred in 29 patients (74%). No imipenem pharmacodynamic parameter was significantly associated with the microbiological success. For amikacin, C1/MIC was significantly higher in the microbiological success group: 26 (1-76) versus 11 (3-26) (P = 0.004). In ICU patients with VAP, classic imipenem pharmacodynamic targets are easily reached with usual dosing regimens. In this context, for amikacin, a higher C1/MIC ratio than previously described might be necessary. © 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.

Biocontrol mechanism by root-associated Bacillus amyloliquefaciens FZB42 – a review

PubMed Central

Chowdhury, Soumitra Paul; Hartmann, Anton; Gao, XueWen; Borriss, Rainer

2015-01-01

Bacillus amyloliquefaciens subsp. plantarum FZB42 is a Gram-positive model bacterium for unraveling plant–microbe interactions in Bacilli. In addition, FZB42 is used commercially as biofertilizer and biocontrol agent in agriculture. Genome analysis of FZB42 revealed that nearly 10% of the FZB42 genome is devoted to synthesizing antimicrobial metabolites and their corresponding immunity genes. However, recent investigations in planta demonstrated that – except surfactin – the amount of such compounds found in vicinity of plant roots is relatively low, making doubtful a direct function in suppressing competing microflora including plant pathogens. These metabolites have been also suspected to induce changes within the rhizosphere microbial community, which might affect environment and plant health. However, sequence analysis of rhizosphere samples revealed only marginal changes in the root microbiome, suggesting that secondary metabolites are not the key factor in protecting plants from pathogenic microorganisms. On the other hand, adding FZB42 to plants compensate, at least in part, changes in the community structure caused by the pathogen, indicating an interesting mechanism of plant protection by beneficial Bacilli. Sub-lethal concentrations of cyclic lipopeptides and volatiles produced by plant-associated Bacilli trigger pathways of induced systemic resistance (ISR), which protect plants against attacks of pathogenic microbes, viruses, and nematodes. Stimulation of ISR by bacterial metabolites is likely the main mechanism responsible for biocontrol action of FZB42. PMID:26284057

Biocontrol mechanism by root-associated Bacillus amyloliquefaciens FZB42 - a review.

PubMed

Chowdhury, Soumitra Paul; Hartmann, Anton; Gao, XueWen; Borriss, Rainer

2015-01-01

Bacillus amyloliquefaciens subsp. plantarum FZB42 is a Gram-positive model bacterium for unraveling plant-microbe interactions in Bacilli. In addition, FZB42 is used commercially as biofertilizer and biocontrol agent in agriculture. Genome analysis of FZB42 revealed that nearly 10% of the FZB42 genome is devoted to synthesizing antimicrobial metabolites and their corresponding immunity genes. However, recent investigations in planta demonstrated that - except surfactin - the amount of such compounds found in vicinity of plant roots is relatively low, making doubtful a direct function in suppressing competing microflora including plant pathogens. These metabolites have been also suspected to induce changes within the rhizosphere microbial community, which might affect environment and plant health. However, sequence analysis of rhizosphere samples revealed only marginal changes in the root microbiome, suggesting that secondary metabolites are not the key factor in protecting plants from pathogenic microorganisms. On the other hand, adding FZB42 to plants compensate, at least in part, changes in the community structure caused by the pathogen, indicating an interesting mechanism of plant protection by beneficial Bacilli. Sub-lethal concentrations of cyclic lipopeptides and volatiles produced by plant-associated Bacilli trigger pathways of induced systemic resistance (ISR), which protect plants against attacks of pathogenic microbes, viruses, and nematodes. Stimulation of ISR by bacterial metabolites is likely the main mechanism responsible for biocontrol action of FZB42.

Surgical Site Infections in Pediatric Spine Surgery: Comparative Microbiology of Patients with Idiopathic and Nonidiopathic Etiologies of Spine Deformity.

PubMed

Maesani, Matthieu; Doit, Catherine; Lorrot, Mathie; Vitoux, Christine; Hilly, Julie; Michelet, Daphné; Vidal, Christophe; Julien-Marsollier, Florence; Ilharreborde, Brice; Mazda, Keyvan; Bonacorsi, Stéphane; Dahmani, Souhayl

2016-01-01

Surgical site infections (SSIs) are a concern in pediatric spine surgery with unusually high rates for a clean surgery and especially for patients with deformity of nonidiopathic etiology. Microbiologic differences between etiologies of spine deformities have been poorly investigated. We reviewed all cases of SSI in spinal surgery between 2007 and 2011. Characteristics of cases and of bacteria according to the etiology of the spine disease were investigated. Of 496 surgeries, we identified 51 SSIs (10.3%) in 49 patients. Staphylococcus aureus was the most frequent pathogen whatever the etiology (n = 31, 61% of infection cases). The second most frequent pathogens vary according to the etiology of the spine deformity. It was Gram-negative bacilli (GNB) in nonidiopathic cases (n = 19, 45% of cases) and anaerobe in idiopathic cases (n = 8, 38% of cases), particularly Gram-positive anaerobic cocci (n = 5, 24% of cases). Infection rate was 6.8% in cases with idiopathic spine disease (n = 21) and 15.9% in cases with nonidiopathic spine disease (n = 30). Nonidiopathic cases were more frequently male with lower weight. American Society of Anesthesiologists score was more often greater than 2, they had more frequently sacral implants and postoperative intensive care unit stay. GNB were significantly associated with a nonidiopathic etiology, low weight, younger age and sacral fusion. SSIs were polymicrobial in 31% of cases with a mean of 1.4 species per infection cases. S. aureus is the first cause of SSI in pediatric spine surgery. However, Gram-positive anaerobic cocci should be taken into account in idiopathic patients and GNB in nonidiopathic patients when considering antibiotic prophylaxis and curative treatment.

Identification of key genes in Gram-positive and Gram-negative sepsis using stochastic perturbation

PubMed Central

Li, Zhenliang; Zhang, Ying; Liu, Yaling; Liu, Yanchun; Li, Youyi

2017-01-01

Sepsis is an inflammatory response to pathogens (such as Gram-positive and Gram-negative bacteria), which has high morbidity and mortality in critically ill patients. The present study aimed to identify the key genes in Gram-positive and Gram-negative sepsis. GSE6535 was downloaded from Gene Expression Omnibus, containing 17 control samples, 18 Gram-positive samples and 25 Gram-negative samples. Subsequently, the limma package in R was used to screen the differentially expressed genes (DEGs). Hierarchical clustering was conducted for the specific DEGs in Gram-negative and Gram-negative samples using cluster software and the TreeView software. To analyze the correlation of samples at the gene level, a similarity network was constructed using Cytoscape software. Functional and pathway enrichment analyses were conducted for the DEGs using DAVID. Finally, stochastic perturbation was used to determine the significantly differential functions between Gram-positive and Gram-negative samples. A total of 340 and 485 DEGs were obtained in Gram-positive and Gram-negative samples, respectively. Hierarchical clustering revealed that there were significant differences between control and sepsis samples. In Gram-positive and Gram-negative samples, myeloid cell leukemia sequence 1 was associated with apoptosis and programmed cell death. Additionally, NADH:ubiquinone oxidoreductase subunit S4 was associated with mitochondrial respiratory chain complex I assembly. Stochastic perturbation analysis revealed that NADH:ubiquinone oxidoreductase subunit B2 (NDUFB2), NDUFB8 and ubiquinol-cytochrome c reductase hinge protein (UQCRH) were associated with cellular respiration in Gram-negative samples, whereas large tumor suppressor kinase 2 (LATS2) was associated with G1/S transition of the mitotic cell cycle in Gram-positive samples. NDUFB2, NDUFB8 and UQCRH may be biomarkers for Gram-negative sepsis, whereas LATS2 may be a biomarker for Gram-positive sepsis. These findings may promote the therapies of sepsis caused by Gram-positive and Gram-negative bacteria. PMID:28714002

Essential Oils and Their Components as Modulators of Antibiotic Activity against Gram-Negative Bacteria

PubMed Central

Aelenei, Petruta; Miron, Anca; Trifan, Adriana; Bujor, Alexandra; Gille, Elvira; Aprotosoaie, Ana Clara

2016-01-01

Gram-negative bacteria cause infections that are difficult to treat due to the emergence of multidrug resistance. This review summarizes the current status of the studies investigating the capacity of essential oils and their components to modulate antibiotic activity against Gram-negative bacteria. Synergistic interactions are particularly discussed with reference to possible mechanisms by which essential oil constituents interact with antibiotics. Special emphasis is given to essential oils and volatile compounds that inhibit efflux pumps, thus reversing drug resistance in Gram-negative bacteria. In addition, indifference and antagonism between essential oils/volatile compounds and conventional antibiotics have also been reported. Overall, this literature review reveals that essential oils and their purified components enhance the efficacy of antibiotics against Gram-negative bacteria, being promising candidates for the development of new effective formulations against Gram-negative bacteria. PMID:28930130

STREPTOCOCCI IN INFECTIOUS (ATROPHIC) ARTHRITIS AND RHEUMATIC FEVER.

PubMed

Nye, R N; Waxelbaum, E A

1930-11-30

The question of the relationship of streptococci to the etiology of infectious arthritis and of rheumatic fever is of the utmost importance. If a streptococcus or group of streptococci could be shown to be associated See PDF for Structure with either disease, some form of specific treatment might be available. The possibility of primary streptococcic infection as the cause of rheumatic fever, and, to a less extent, of acute infectious arthritis would seem to be a reasonable conjecture because of the frequency of associated throat, sinus or other focal infection. To consider that these same streptococci remain in or about the affected joint and to such an extent that they are found in the blood stream in cases of chronic infectious arthritis of years' duration demands a rather unique conception. Recent investigative work has certainly tended to confirm the importance of streptococci in these diseases, but, if all the published reports are considered as a group, one can not help being impressed with the inconsistency and peculiarities of the findings. In blood cultures from cases of rheumatic fever Clawson (7) recovered Streptococcus viridans, Small (8) and Birkhaug (9) non-hemolytic (gamma type) streptococci, and Cecil et al. (3) Streptococcus viridans, rarely hemolytic and non-hemolytic streptococci. In blood cultures from cases of infectious arthritis Cecil et al. (2) recovered attenuated hemolytic streptococci and occasionally Streptococcus viridans, non-hemolytic streptococci and diphtheroids and Margolis and Dorsey (10) green-producing and indifferent streptococci and diphtheroids, whereas from synovial fluids and regional lymph nodes Forkner, Shands and Poston (4) and Poston (5) obtained Streptococcus viridans and Margolis and Dorsey (11) recovered green-producing and indifferent streptococci and diphtheroids from epiphyseal marrows, bones and synovial membranes. On the other hand, Jordan (12) and Nye and Seegal (13) and the work reported in this paper have failed to confirm these findings. If a streptococcus is the infective agent, it is difficult to explain why the organisms recovered so consistently by certain groups of investigators are so different and why the findings of other groups are entirely negative. The chances of contamination, even with the most careful manipulation, are extremely favorable when using a cultural technique which demands subculturing a fluid medium every 3 to 5 days for a period of 4 to 6 weeks. The question arises as to what organisms should be considered as contaminants. Cultures found to contain Staphylococcus albus have never been judged significant. Margolis and Dorsey (11) have excluded Gram-positive bacilli from their series, but have included diphtheroids. Cecil et al. (2) reported the recovery of diphtheroids and Micrococcus zymogenes from blood cultures which they did not consider contaminated. Jordan (12) recovered short Gram-negative bacilli and Gram-positive bacilli and questioned their importance. The occurrence of such bacteria would appear to be not unlike that of the non-hemolytic (gamma type) streptococci reported by Small (7), Birkhaug (9) and, rarely, by Cecil et al. (2) and Margolis and Dorsey (10,11). In the work reported in this paper and in previously published work (13) with blood cultures from cases of rheumatic fever, staphylococci, Gram-positive bacilli and diplococci and diphtheroids have been isolated from a certain number of the cultures. Since these were found, as a rule, in only 1 of 2 or more cultures from the same blood and since such organisms occurred about as frequently in cultures from control cases, they have been considered merely contaminants. The findings in the 9 cases of arthritis with positive blood cultures reported by Margolis and Dorsey (10) are certainly far from convincing. These authors found green-producing streptococci in 5 of the 9 cases, but duplicate cultures on the same blood, with one exception, were always negative and the two repeat cultures were negative. Furthermore, in the 3 cases yielding indifferent streptococci in one subculture all subsequent subcultures were negative. In the work reported in this paper a green-producing and a non-hemolytic streptococcus were isolated from blood cultures, but a Gram-positive diplococcus and a Gram-positive bacillus, respectively, were recovered from the same cultures. A consideration of the above would seem to point to the probability, even certainty, of streptococci occurring in some cultures as contaminants; and the work of Olitsky and Long (14) and Long, Olitsky and Stewart (15) has clearly demonstrated that the air contamination of cultures of ground material with non-hemolytic green-producing streptococci can occur just as easily as with diphtheroids and staphylococci. It is obvious, in such cases, that the types of organisms recovered are dependent on the flora of the air of the laboratory or of the throat of the laboratory worker, and this point may well explain the differences in the cultural findings under consideration. A number of years ago numerous articles (16-19) appeared relative to the bacteriologic flora of lymph nodes, particularly those from cases of Hodgkin's disease, and it is interesting to note that the organisms recovered were quite similar to those which have been recovered from the blood and tissues of cases of arthritis and rheumatic fever, with the exception that the diphtheroids were, at that time, much more in prominence. Regardless of elaborate serological studies and animal experiments, streptococci must be recovered consistently by several groups of laboratory workers before their etiologic rôle in chronic arthritis and rheumatic fever can be accepted. Duplicate cultures and repeat cultures should yield the same organism in a generous percentage of cases and cultures from cases of other diseases or from normal persons should be negative. Positive cultures from duplicate cultures opened only at the time when the first culture showed growth would make the findings more significant than if the whole series were subcultured every 3 to 5 days. The work of Cecil, Nicholls and Stainsby on the bacteriology of the blood and joints in chronic arthritis and rheumatic fever has apparently been carried out most carefully and thoroughly and their results are very consistent and convincing. In spite of attempts to follow their methods in the selection of patients and in cultural technique, the results on the relatively small series of cases which are reported here fail to confirm their findings.

Clinicomicrobiological profile of endophthalmitis: A 10 year experience in a Tertiary Care Center in North India.

PubMed

Satpathy, Gita; Nayak, Niranjan; Wadhwani, Meenakshi; Venkwatesh, Pradeep; Kumar, Atul; Sharma, Yograj; Sreenivas, Vishnu

2017-01-01

To determine the clinicomicrobiological profile of infectious agents and their antibiotic susceptibility in different type of endophthalmitis. A retrospective review of clinical and microbiological records from January 2001 to December 2010, was performed in 1110 patients diagnosed with different type of endophthalmitis (postoperative, posttraumatic, endogenous and post keratitis) to record the demographic details, clinical presentations; microbiological agents isolated with their antimicrobial sensitivity pattern. Antimicrobial susceptibility testing for various culture positive isolates (bacterial/fungal) was performed by the disc diffusion technique. Out of the 1110 intra-ocular specimens processed, 384 (34.6%) were positive for bacteria. S epidermidis was the most predominant isolate accounting for 42.7% of all bacteria obtained, followed by Pseudomonas aeruginosa (24.5%). Besides Pseudomonas, Acinetobacter spp. were the next common gram negative bacilli detected (8.3%) followed by Klebsiella, E. coli, Enterobacter and Alkaligenes in 2.6%, 0.8%, 0.8% and 0.5% cases respectively. The predominant fungal species were Aspergillus spp., in 36.1%, followed by Fusarium spp. in 26.4% cases. Overall susceptibility pattern in our study showed that gram positive bacteria were most susceptible to glycopeptides like vancomycin (80-100%) and fluoroquinolones (87-91%). The sensitivity pattern of gram negative organisms like Pseudomonas and Klebsiella towards fluoroquinolones ranged between 61% - 82%. S epidermidis was the most common bacteria isolated in postoperative and posttraumatic endophthalmitis, Pseudomonas aeruginosa was the most common bacterial isolated in posttraumatic endophthalmitisAmongst fungi Aspergillus was the most common organism.

The effects of antiperspirant on the perineal skin flora of patients with spinal cord injury.

PubMed

Montgomerie, J Z; Gilmore, D S; Graham, I E; Schick, D G

1988-07-01

The relationship of pH and moisture to Pseudomonas aeruginosa and Klebsiella pneumoniae colonization of the perineal skin was studied in male patients with spinal cord injury. The increased pH of the perineal skin was significantly associated with the presence of P. aeruginosa but not other bacterial species. No correlation between colonization and moisture or pH and moisture was found. An antiperspirant produced a significant reduction in the number of total aerobic bacteria, total gram-negative bacilli, P. aeruginosa and K. pneumoniae over a 24-h period. Long-term use of the antiperspirant for 10 days did not alter the persistence of P. aeruginosa of the same serotypes on the perineum.

Two Cases of Bacteremia Due to Roseomonas mucosa.

PubMed

Kim, Yu Kyung; Moon, Jung Suk; Song, Kyung Eun; Lee, Won Kil

2016-07-01

Roseomonas is a genus of pink-pigmented nonfermentative bacilli. These slow-growing, gram-negative cocobacilli form pink-colored colonies on sheep blood agar. They differ from other pink-pigmented nonfermenters, including Methylobacterium, in morphology, biochemical characteristics, and DNA sequence. Roseomonas strains are rarely isolated in clinical laboratories; therefore, we report two cases in order to improve our ability to identify these pathogens. We isolated two strains of Roseomonas mucosa from the venous blood cultures of two patients, an 84-yr-old woman with common bile duct obstruction and a 17-yr-old male with acute myeloid leukemia who had an indwelling central-venous catheter for chemotherapy. The isolated strains were confirmed as R. mucosa by 16S rRNA sequencing.

Two Cases of Bacteremia Due to Roseomonas mucosa

PubMed Central

Kim, Yu Kyung; Moon, Jung Suk; Song, Kyung Eun

2016-01-01

Roseomonas is a genus of pink-pigmented nonfermentative bacilli. These slow-growing, gram-negative cocobacilli form pink-colored colonies on sheep blood agar. They differ from other pink-pigmented nonfermenters, including Methylobacterium, in morphology, biochemical characteristics, and DNA sequence. Roseomonas strains are rarely isolated in clinical laboratories; therefore, we report two cases in order to improve our ability to identify these pathogens. We isolated two strains of Roseomonas mucosa from the venous blood cultures of two patients, an 84-yr-old woman with common bile duct obstruction and a 17-yr-old male with acute myeloid leukemia who had an indwelling central-venous catheter for chemotherapy. The isolated strains were confirmed as R. mucosa by 16S rRNA sequencing. PMID:27139611

Resistance profile for pathogens causing urinary tract infection in a pediatric population, and antibiotic treatment response at a university hospital, 2010-2011.

PubMed

Vélez Echeverri, Catalina; Serna-Higuita, Lina María; Serrano, Ana Katherina; Ochoa-García, Carolina; Rojas Rosas, Luisa; María Bedoya, Ana; Suárez, Margarita; Hincapié, Catalina; Henao, Adriana; Ortiz, Diana; Vanegas, Juan José; Zuleta, John Jairo; Espinal, David

2014-01-01

Urinary tract infection (UTI) is one of the most common bacterial infections in childhood and causes acute and chronic morbidity and long-term hypertension and chronic kidney disease. To describe the demographic characteristics, infectious agents, patterns of antibiotic resistance, etiologic agent and profile of susceptibility and response to empirical treatment of UTI in a pediatric population. This is a descriptive, retrospective study. Included in the study were 144 patients, 1:2.06 male to female ratio. The most common symptom was fever (79.9%) and 31.3% had a history of previous UTI. 72.0% of the patients had positive urine leukocyte count (>5 per field), urine gram was positive in 85.0% of samples and gram negative bacilli accounted for 77.8% for the total pathogens isolated. The most frequent uropathogens isolated were Escherichia coli and Klebsiella pneumoniae. Our E.coli isolates had a susceptibility rate higher than 90% to most of the antibiotics used, but a resistance rate of 42.6% to TMP SMX and 45.5% to ampicillin sulbactam. 6.3% of E. coli was extended-spectrum beta-lactamases producer strains. The most frequent empirical antibiotic used was amikacin, which was used in 66.0% of the patients. 17 of 90 patients who underwent voiding cistouretrography (VCUG) had vesicoureteral reflux. This study revealed that E. coli was the most frequent pathogen of community acquired UTI. We found that E. coli and other uropathogens had a high resistance rate against TMP SMX and ampicillin sulbactam. In order to ensure a successful empirical treatment, protocols should be based on local epidemiology and susceptibility rates.

Resistance profile for pathogens causing urinary tract infection in a pediatric population, and antibiotic treatment response at a University Hospital, 2010-2011

PubMed Central

Vélez Echeverri, Catalina; Serrano, Ana Katherina; Ochoa-García, Carolina; Rojas Rosas, Luisa; María Bedoya, Ana; Suárez, Margarita; Hincapié, Catalina; Henao, Adriana; Ortiz, Diana; Vanegas, Juan José; Zuleta, John Jairo; Espinal, David

2014-01-01

Introduction: Urinary tract infection (UTI) is one of the most common bacterial infections in childhood and causes acute and chronic morbidity and long-term hypertension and chronic kidney disease. Objectives: To describe the demographic characteristics, infectious agents, patterns of antibiotic resistance, etiologic agent and profile of susceptibility and response to empirical treatment of UTI in a pediatric population. Methods: This is a descriptive, retrospective study. Results: Included in the study were 144 patients, 1:2.06 male to female ratio. The most common symptom was fever (79.9%) and 31.3% had a history of previous UTI. 72.0% of the patients had positive urine leukocyte count (>5 per field), urine gram was positive in 85.0% of samples and gram negative bacilli accounted for 77.8% for the total pathogens isolated. The most frequent uropathogens isolated were Escherichia coli and Klebsiella pneumoniae. Our E.coli isolates had a susceptibility rate higher than 90% to most of the antibiotics used, but a resistance rate of 42.6% to TMP SMX and 45.5% to ampicillin sulbactam. 6.3% of E. coli was extended-spectrum beta-lactamases producer strains. The most frequent empirical antibiotic used was amikacin, which was used in 66.0% of the patients. 17 of 90 patients who underwent voiding cistouretrography (VCUG) had vesicoureteral reflux. Conclusion: This study revealed that E. coli was the most frequent pathogen of community acquired UTI. We found that E. coli and other uropathogens had a high resistance rate against TMP SMX and ampicillin sulbactam. In order to ensure a successful empirical treatment, protocols should be based on local epidemiology and susceptibility rates. PMID:24970958

EFFECT OF SILICATE ON GRAM STAINING AND VIABILITY OF PNEUMOCOCCI AND OTHER BACTERIA

PubMed Central

MacLeod, Colin M.; Roe, Amy S.

1956-01-01

Application of silicate solutions to living or heat-killed pneumococci and to certain "viridans" streptococci causes their conversion from a Gram-positive to a Gram-negative state. The original staining properties can be restored by suspending the silicate-treated bacteria in alkaline solutions of various salts but not by simple washing in water. Living pneumococci and the strains of streptococci whose staining properties are similarly affected are killed when suspended in silicate solutions. In other Gram-positive species silicate causes conversion to Gram negativity but restoration to positivity occurs upon washing in water. In a third group of Gram-positive organisms silicate has no effect on the Gram reaction. The viability of organisms in these two groups is unaffected by silicate under the conditions employed. No effect on staining or viability of Gram-negative bacteria has been observed. The effects of silicate on staining and viability are inhibited by nutrient broth or whole serum but not by purified serum albumin. Lecithin, choline, and other substituted ammonium compounds also inhibit the effects of silicate on pneumococci. PMID:13306854

Antimicrobial resistance in Gram-positive bacteria from Timorese River Buffalo (Bubalus bubalis) skin microbiota.

PubMed

Oliveira, Manuela; Monteiro, José L; Rana, Sílvia; Vilela, Cristina L

2010-06-01

The Timorese River Buffalo (Bubalus bubalis) plays a major role in the East Timor economy, as it is an important source of animal protein in human nutrition. They are widely spread throughout the country and are in direct contact with the populations. In spite of this proximity, information on their microbiota is scarce. This work aimed at characterizing the skin microbiota of the East Timorese River Buffalo and its antimicrobial resistance profile. Skin swab samples were taken from 46 animals in surveys conducted in three farms located in "Suco de Nairete", Lospalos district, during July and August 2006. Bacteria were isolated and identified according to conventional microbiological procedures. A total of 456 isolates were obtained, including Gram-positive (n = 243) and Gram-negative (n = 213) bacteria. Due to their importance as potential pathogens and as vehicles for antimicrobial resistance transmission, Gram-positive cocci (n = 27) and bacilli (n = 77) isolates were further characterized, and their antimicrobial resistance profile determined by the disk diffusion method according to the Clinical and Laboratory Standards Institute guidelines. This study shows the high bacterial diversity of B. bubalis skin microbiota, representing an important first step towards understanding its importance and epidemiologic role in animal health. It also points out the potential role of these animals as vectors of antimicrobial resistant bacteria dissemination and the importance of antimicrobial resistance monitoring in developing countries.

Microbial investigation of biofilms recovered from endotracheal tubes using sonication in intensive care unit pediatric patients.

PubMed

Ferreira, Thiago de Oliveira; Koto, Rafael Yoshio; Leite, Gabriel Fialkovitz da Costa; Klautau, Giselle Burlamaqui; Nigro, Stanley; Silva, Cely Barreto da; Souza, Ana Paula Idalgo da Fonseca; Mimica, Marcelo Jenne; Cesar, Regina Grigolli; Salles, Mauro José Costa

2016-01-01

To compare cultured microorganisms identified on endotracheal tubes biofilms through sonication technique with traditional tracheal aspirate collected at extubation of pediatric intensive care unit patients. Demographic and epidemiological data were analyzed to identify factors possibly related with the microbiological profile of the two collection methods. Associations between categorical and continuous variables were analyzed using the chi-square or Fisher's exact test, or Student's t test. p-Value <0.05 were considered significant. Thirty endotracheal tubes and tracheal aspirates samples from 27 subjects were analyzed. Only one patient presented the clinical diagnosis of ventilator-associated pneumonia. Overall, 50% of bacteria were Gram-negative bacilli, followed by Gram-positive bacteria in 37%, and fungi in 10%. No statistically significant difference on the distribution of Gram-positive or Gram-negative bacteria (p=0.996), and fungi (p=0.985) were observed between the collection methods. Pseudomonas spp. was the most frequent microorganism identified (23.8%), followed by Streptococcus spp. (18.5%), Acinetobacter spp. (15.9%), coagulase-negative staphylococci (11.2%), and Klebsiella spp. (8.6%). Concordant results between methods amounted to 83.3%. Pseudomonas aeruginosa and Acinetobacter baumannii showed carbapenem resistance in 50% and 43.7% of the isolates, respectively. In general, cultures after endotracheal tubes sonication (non-centrifuged sonication fluid and centrifuged sonication fluid) yielded bacteria with higher rates of antimicrobial resistance compared to tracheal aspirates cultures. Additionally, in 12 subjects (40%), we observed discrepancies regarding microbiologic profiles of cultures performed using the collection methods. Our study demonstrated that sonication technique can be applied to ET biofilms to identify microorganisms attached to their surface with a great variety of species identified. However, we did not find significant differences in comparison with the traditional tracheal aspirate culture approach. Copyright © 2016 Sociedade Brasileira de Infectologia. Published by Elsevier Editora Ltda. All rights reserved.

Effect of extremely low frequency electromagnetic fields on growth rate and morphology of bacteria.

PubMed

Inhan-Garip, Ayse; Aksu, Burak; Akan, Zafer; Akakin, Dilek; Ozaydin, A Nilufer; San, Tangul

2011-12-01

To determine the effect of extremely low frequency (<300 Hz) electromagnetic fields (ELF-EMF) on the growth rate of Gram-positive and Gram-negative bacteria and to determine any morphological changes that might have been caused by ELF-EMF. Six bacterial strains, three Gram-negative and three Gram-positive were subjected to 50 Hz, 0.5 mT ELF-EMF for 6 h. To determine growth rate after ELF-EMF application, bacteria exposed to ELF-EMF for 3 h were collected, transferred to fresh medium and cultured without field application for another 4 h. Growth-rate was determined by optical density (OD) measurements made every hour. Morphological changes were determined with Transmission electron microscopy (TEM) for two gram-negative and two gram-positive strains collected after 3 h of field application. A decrease in growth rate with respect to control samples was observed for all strains during ELF-EMF application. The decrease in growth-rate continued when exposed bacteria were cultured without field application. Significant ultrastructural changes were observed in all bacterial strains, which were seen to resemble the alterations caused by cationic peptides. This study shows that ELF-EMF induces a decrease in growth rate and morphological changes for both Gram-negative and Gram-positive bacteria.

Comparison of two commercial carbapenemase gene confirmatory assays in multiresistant Enterobacteriaceae and Acinetobacter baumannii-complex.

PubMed

Rösner, Stephan; Gehlweiler, Kevin; Küsters, Uta; Kolbert, Mathias; Hübner, Kirsten; Pfennigwerth, Niels; Mack, Dietrich

2018-01-01

Multidrug-resistant Gram-negative bacilli (MDR-GNB) producing carbapenemases are increasing at an alarming speed. Rapid confirmation of carbapenemase type will be an important diagnostic step in clinical microbiology laboratories not only to reduce the risk of transmissions but also for optimising antibiotic therapy in the future. We compared diagnostic reliability of two commercially available molecular assays (Check-Direct CPE vs. AID line probe assay) for detection and typing of carbapenemase genes in 80 well-characterized isolates of MDR-GNB. Respective strains were isolated in various clinical specimens at our clinical microbiology laboratory. The reference standard included confirmation of carbapenemase-production at the molecular level at the German National Reference Laboratory for Multidrug-resistant Gram-negative bacteria (Ruhr-University Bochum, Germany). 53 Enterobacteriaceae and 27 members of the A. baumannii-complex were used in this study. The tested assays appeared highly reliable to confirm carbapenemase-producing Enterobacteriaceae (CPE) with respective sensitivities of 97.7%, but are currently unsuitable for analysis of members of the A. baumannii-complex. Both assays are easy to perform and rapid tools for confirmation and typing of the most common carbapenemase genes in Enterobacteriaceae. Implementation should be possible for any clinical microbiology laboratory with Check-Direct CPE being easier to handle and having less technological requirements.

Symptomatic Peripheral Mycotic Aneurysms Due to Infective Endocarditis

PubMed Central

González, Isabel; Sarriá, Cristina; López, Javier; Vilacosta, Isidre; San Román, Alberto; Olmos, Carmen; Sáez, Carmen; Revilla, Ana; Hernández, Miguel; Caniego, Jose Luis; Fernández, Cristina

2014-01-01

Abstract Peripheral mycotic aneurysms (PMAs) are a relatively rare but serious complication of infective endocarditis (IE). We conducted the current study to describe and compare the current epidemiologic, microbiologic, clinical, diagnostic, therapeutic, and prognostic characteristics of patients with symptomatic PMAs (SPMAs). A descriptive, comparative, retrospective observational study was performed in 3 tertiary hospitals, which are reference centers for cardiac surgery. From 922 definite IE episodes collected from 1996 to 2011, 18 patients (1.9%) had SPMAs. Because all SPMAs developed in left-sided IE, we performed a comparative study between 719 episodes of left-sided IE without SPMAs and 18 episodes with SPMAs. We found a higher frequency of intravenous drug abuse, native valve IE, intracranial bleeding, septic emboli, multiple embolisms, and IE diagnostic delay >30 days in patients with SPMAs than in patients without SPMAs. The causal microorganisms were gram-positive cocci (n =10), gram-negative bacilli (n = 2), gram-positive bacilli (n = 3), Bartonella henselae (n = 1), Candida albicans (n = 1), and negative culture (n = 1). The median IE diagnosis delay was 15 days (interquartile range [IQR], 13–33 d) in the case of high-virulence microorganisms versus 45 days (IQR, 30–240 d) in the case of low- to medium-virulence microorganisms. Twelve SPMAs were intracranial and 6 were extracranial. In 10 cases (8 intracranial and 2 extracranial), SPMAs were the initial presentation of IE; the remaining cases developed symptoms during or after finishing parenteral antibiotic treatment. The initial diagnosis of intracranial SPMAs was made by computed tomography (CT) or magnetic resonance imaging in 6 unruptured aneurysms and by angiography in 6 ruptured aneurysms. The initial test in extracranial SPMAs was Doppler ultrasonography in limbs, CT in liver, and coronary angiography in heart. Four (3 intracranial, 1 extracranial) of 7 (6 intracranial, 1 extracranial) patients treated only with antibiotics died. Surgical resection was performed in 7 (3 intracranial, 4 extracranial) and endovascular repair in 4 (3 intracranial, 1 extracranial) patients; all of them survived. In conclusion, we found that SPMAs were a rare complication of IE that developed only in left-sided IE, and especially in native valves. Intracranial hemorrhage, embolism, multiple embolisms, and diagnostic delay of IE were more common in patients with SPMAs. The microbiologic profile was diverse, but microorganisms of low-medium virulence were predominant, and had a greater delayed diagnosis of IE than those caused by microorganisms of high virulence. SPMAs were often the initial presentation of IE. The most common location of SPMAs was intracranial. Noninvasive radiologic imaging techniques were the initial imaging test in intracranial unruptured SPMAs and in most extracranial SPMAs. Surgical and endovascular treatments were safe and effective. Endovascular treatment could be the first line of treatment in selected cases. Mortality was high in those cases treated only with antibiotics. PMID:24378742

A Comparative Evaluation on Antimicrobial Effect of Honey, Neem Leaf Extract and Sodium Hypochlorite as Intracanal Irrigant: An Ex-Vivo Study

PubMed Central

Narayanan, Retna Kumari; Vadakkepurayil, Kannan

2016-01-01

Introduction The major determinant of the success of root canal treatment depends on meticulous disinfection of the root canal using intracanal irrigants. The most commonly used root canal irrigant is sodium hypochlorite which has disadvantages of cytotoxicity and unpleasant taste. So there is a need to identify a more biocompatible root canal irrigant. Aim The aim of this ex-vivo study was to evaluate the efficacy of 40% honey, 100% neem leaf extract and 5.25% sodium hypochlorite as an intracanal irrigant against the isolated microorganisms from infected root canal. Materials and Methods The samples were collected from infected root canals of 60 primary molar teeth indicated for pulpectomy. Alpha hemolytic Streptococci, gram negative bacilli, Candida, Staphylococci, Lactobacilli, Enterococci, Spore bearing gram positive bacilli and Micrococci were the microorganisms isolated from the samples. The zone of inhibition against the microbial growth was measured by agar well diffusion method. Statistical analysis was done by Repeated Analysis of Variance (ANOVA) and Bonferroni method. Results Statistical analysis showed that the means of the zones of inhibition measured in this study were 18.56mm, 2.09mm and 1.62mm for sodium hypochlorite, 100% neem leaf extract and 40% honey respectively. The significance was greater between sodium hypochlorite and the other two agents as p-value was <0.001. Conclusion The results indicated that 5.25% sodium hypochlorite is more effective as root canal irrigant when compared with 100% neem leaf extract and 40% honey. It was also observed that 100% neem leaf extract has greater antimicrobial effect than 40% honey. PMID:27656571

Combination Therapy for Treatment of Infections with Gram-Negative Bacteria

PubMed Central

Cosgrove, Sara E.; Maragakis, Lisa L.

2012-01-01

Summary: Combination antibiotic therapy for invasive infections with Gram-negative bacteria is employed in many health care facilities, especially for certain subgroups of patients, including those with neutropenia, those with infections caused by Pseudomonas aeruginosa, those with ventilator-associated pneumonia, and the severely ill. An argument can be made for empiric combination therapy, as we are witnessing a rise in infections caused by multidrug-resistant Gram-negative organisms. The wisdom of continued combination therapy after an organism is isolated and antimicrobial susceptibility data are known, however, is more controversial. The available evidence suggests that the greatest benefit of combination antibiotic therapy stems from the increased likelihood of choosing an effective agent during empiric therapy, rather than exploitation of in vitro synergy or the prevention of resistance during definitive treatment. In this review, we summarize the available data comparing monotherapy versus combination antimicrobial therapy for the treatment of infections with Gram-negative bacteria. PMID:22763634

Associations Between Enteral Colonization With Gram-Negative Bacteria and Intensive Care Unit-Acquired Infections and Colonization of the Respiratory Tract.

PubMed

Frencken, Jos F; Wittekamp, Bastiaan H J; Plantinga, Nienke L; Spitoni, Cristian; van de Groep, Kirsten; Cremer, Olaf L; Bonten, Marc J M

2018-02-01

Enteral and respiratory tract colonization with gram-negative bacteria may lead to subsequent infections in critically ill patients. We aimed to clarify the interdependence between gut and respiratory tract colonization and their associations with intensive care unit (ICU)-acquired infections in patients receiving selective digestive tract decontamination (SDD). Colonization status of the rectum and respiratory tract was determined using twice-weekly microbiological surveillance in mechanically ventilated subjects receiving SDD between May 2011 and June 2015 in a tertiary medical-surgical ICU in the Netherlands. Acquisition of infections was monitored daily by dedicated observers. Marginal structural models were used to determine the associations between gram-negative rectal colonization and respiratory tract colonization, ICU-acquired gram-negative infection, and ICU-acquired gram-negative bacteremia. Among 2066 ICU admissions, 1157 (56.0%) ever had documented gram-negative carriage in the rectum during ICU stay. Cumulative incidences of ICU-acquired gram-negative infection and bacteremia were 6.0% (n = 124) and 2.1% (n = 44), respectively. Rectal colonization was an independent risk factor for both respiratory tract colonization (cause-specific hazard ratio [CSHR], 2.93 [95% confidence interval {CI}, 2.02-4.23]) and new gram-negative infection in the ICU (CSHR, 3.04 [95% CI, 1.99-4.65]). Both rectal and respiratory tract colonization were associated with bacteremia (CSHR, 7.37 [95% CI, 3.25-16.68] and 2.56 [95% CI, 1.09-6.03], respectively). Similar associations were observed when Enterobacteriaceae and glucose nonfermenting gram-negative bacteria were analyzed separately. Gram-negative rectal colonization tends to be stronger associated with subsequent ICU-acquired gram-negative infections than gram-negative respiratory tract colonization. Gram-negative rectal colonization seems hardly associated with subsequent ICU-acquired gram-negative respiratory tract colonization. © The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.

Accuracy of the cytopathology, bacterioscopy, and vaginal flora culture.

PubMed

Almeida, P Q; Pereira, M A P; Palomo, F S; Okazaki, C; Schimidt, M A; Speck, N M G; Ribalta, J C L

2013-01-01

An over-population of vaginal microorganisms causing inflammatory processes renders it difficult to properly assess the cytopathological exam that aims to screen precedent cervical lesions. On the contrary, the occurrence of the microbial flora saprophyte does not influence correct cythodiagnosis. To assess the composition of vaginal tract aerobic microorganisms of asymptomatic women in menacme and post-menopause, and to analyze the accuracy of cytopathologic, bacterioscopic exams, and culturing of the flora. The women were first submitted to a focused anamnestic interrogatory and then submitted to gynecological exam. A sample of the vaginal fluid was collected with a culture swab and a smear was made on two glass slides for stained bacterioscopic exam (GRAM). The collection of material was then compiled in a cytopathologic smear analysis. All women signed the free and informed consent letter and the project was approved by the Ethics Research Board of Hospital São Paulo - UNIFESP. Bacterioscopy and culture proved to be better than the cytopathologic exam in featuring the bacilli and cocci. The bacterioscopy provided a better detection of the presence of bacilli (p < 0.001); no statistical difference was seen between both exams with respect to the detected cocci. The beta-hemolytic Streptococcus group was of significance in post-menopausal women (p < 0.05). In this study, the bacterioscopic and culture exams of the vaginal fluid were more effective in assessing the vaginal flora and in the detection of bacilli, compared to the cytopathological exam.

Performance of Two Resin-Containing Blood Culture Media in Detection of Bloodstream Infections and in Direct Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (MALDI-TOF MS) Broth Assays for Isolate Identification: Clinical Comparison of the BacT/Alert Plus and Bactec Plus Systems

PubMed Central

Fiori, Barbara; D'Inzeo, Tiziana; Di Florio, Viviana; De Maio, Flavio; De Angelis, Giulia; Giaquinto, Alessia; Campana, Lara; Tanzarella, Eloisa; Tumbarello, Mario; Antonelli, Massimo; Spanu, Teresa

2014-01-01

We compared the clinical performances of the BacT/Alert Plus (bioMérieux) and Bactec Plus (Becton Dickinson) aerobic and anaerobic blood culture (BC) media with adsorbent polymeric beads. Patients ≥16 years old with suspected bloodstream infections (BSIs) were enrolled in intensive care units and infectious disease wards. A single 40-ml blood sample was collected from each and used to inoculate (10 ml/bottle) one set of BacT/Alert Plus cultures and one set of Bactec Plus cultures, each set consisting of one aerobic and one anaerobic bottle. Cultures were incubated ≤5 days in the BacT/Alert 3D and Bactec FX instruments, respectively. A total of 128 unique BSI episodes were identified based on the recovery of clinically significant growth in 212 aerobic cultures (106 BacT/Alert and 106 Bactec) and 151 anaerobic cultures (82 BacT/Alert and 69 Bactec). The BacT/Alert aerobic medium had higher recovery rates for Gram-positive cocci (P = 0.024), whereas the Bactec aerobic medium was superior for recovery of Gram-negative bacilli (P = 0.006). BacT/Alert anaerobic medium recovery rates exceeded those of the Bactec anaerobic medium for total organisms (P = 0.003), Gram-positive cocci (P = 0.013), and Escherichia coli (P = 0.030). In terms of capacity for diagnosing the 128 septic episodes, the BacT/Alert and Bactec sets were comparable, although the former sets diagnosed more BSIs caused by Gram-positive cocci (P = 0.008). They also allowed earlier identification of coagulase-negative staphylococcal growth (mean, 2.8 h; P = 0.003) and growth in samples from patients not on antimicrobial therapy that yielded positive results (mean, 1.3 h; P < 0.001). Similarly high percentages of microorganisms in BacT/Alert and Bactec cultures (93.8% and 93.3%, respectively) were identified by direct matrix-assisted laser desorption ionization–time of flight mass spectrometry assay of BC broths. The BacT/Alert Plus media line appears to be a reliable, timesaving tool for routine detection of BSIs in the population we studied, although further studies are needed to evaluate their performance in other settings. PMID:25031441

Battacin (Octapeptin B5), a New Cyclic Lipopeptide Antibiotic from Paenibacillus tianmuensis Active against Multidrug-Resistant Gram-Negative Bacteria

PubMed Central

Qian, Chao-Dong; Teng, Yi; Zhao, Wen-Peng; Li, Ou; Fang, Sheng-Guo; Huang, Zhao-Hui; Gao, Hai-Chun

2012-01-01

Hospital-acquired infections caused by drug-resistant bacteria are a significant challenge to patient safety. Numerous clinical isolates resistant to almost all commercially available antibiotics have emerged. Thus, novel antimicrobial agents, specifically those for multidrug-resistant Gram-negative bacteria, are urgently needed. In the current study, we report the isolation, structure elucidation, and preliminary biological characterization of a new cationic lipopeptide antibiotic, battacin or octapeptin B5, produced from a Paenibacillus tianmuensis soil isolate. Battacin kills bacteria in vitro and has potent activity against Gram-negative bacteria, including multidrug-resistant and extremely drug-resistant clinical isolates. Hospital strains of Escherichia coli and Pseudomonas aeruginosa are the pathogens most sensitive to battacin, with MICs of 2 to 4 μg/ml. The ability of battacin to disrupt the outer membrane of Gram-negative bacteria is comparable to that of polymyxin B, the last-line therapy for infections caused by antibiotic-resistant Gram-negative bacteria. However, the capacity of battacin to permeate bacterial plasma membranes is less extensive than that of polymyxin B. The bactericidal kinetics of battacin correlate with the depolarization of the cell membrane, suggesting that battacin kills bacteria by disrupting the cytoplasmic membrane. Other studies indicate that battacin is less acutely toxic than polymyxin B and has potent in vivo biological activity against E. coli. Based on the findings of the current study, battacin may be considered a potential therapeutic agent for the treatment of infections caused by antibiotic-resistant Gram-negative bacteria. PMID:22183171

Permeability barrier of Gram-negative cell envelopes and approaches to bypass it

DOE PAGES

Zgurskaya, Helen I.; López, Cesar A.; Gnanakaran, Sandrasegaram

2015-09-18

Gram-negative bacteria are intrinsically resistant to many antibiotics. Species that have acquired multidrug resistance and cause infections that are effectively untreatable present a serious threat to public health. The problem is broadly recognized and tackled at both the fundamental and applied levels. This article summarizes current advances in understanding the molecular bases of the low permeability barrier of Gram-negative pathogens, which is the major obstacle in discovery and development of antibiotics effective against such pathogens. Gaps in knowledge and specific strategies to break this barrier and to achieve potent activities against difficult Gram-negative bacteria are also discussed.

Permeability barrier of Gram-negative cell envelopes and approaches to bypass it

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zgurskaya, Helen I.; López, Cesar A.; Gnanakaran, Sandrasegaram

Gram-negative bacteria are intrinsically resistant to many antibiotics. Species that have acquired multidrug resistance and cause infections that are effectively untreatable present a serious threat to public health. The problem is broadly recognized and tackled at both the fundamental and applied levels. This article summarizes current advances in understanding the molecular bases of the low permeability barrier of Gram-negative pathogens, which is the major obstacle in discovery and development of antibiotics effective against such pathogens. Gaps in knowledge and specific strategies to break this barrier and to achieve potent activities against difficult Gram-negative bacteria are also discussed.

The role of chiropractic adjusting tables as reservoirs for microbial diseases.

PubMed

Bifero, Antonio E; Prakash, Jaya; Bergin, Jeff

2006-04-01

Our goal was to enumerate the microbial flora on the headrest, armrest, and thoracic portion of chiropractic adjusting tables to determine the presence of pathogenic microorganisms and identify the potential for nosocomial transmission. Defined portions of the headrest, armrest, and thoracoabdominal pieces from 9 chiropractic adjusting tables were randomly sampled using the ACT II culture transport system (Remel, Lenexa, KS). Samples were incubated using standard culture techniques for bacteriology and mycology that included 15 mL of heart infusion agar at 35 degrees C for 24 hours and 15 mL of Sabouraud dextrose agar at 30 degrees C up to 7 days. Identification of microorganisms by biochemical analysis yielded a wide variety of gram-positive (G+) and gram-negative (G-) cocci and bacilli as well as standard fungi. Many were of the genus Staphylococcus. Methicillin-resistant S. aureus (MRSA) isolates were recovered from 2 separate tables. This study supports a growing consensus that those outpatient facilities that do not adhere to strict infection control protocols may become ecological reservoirs of potentially harmful human pathogens.

Levofloxacin for the treatment of community-acquired pneumonia.

PubMed

Lynch, Joseph P; File, Thomas M; Zhanel, George G

2006-10-01

New respiratory fluoroquinolones (FQs), such as levofloxacin, offer many improved qualities over older agents, such as ciprofloxacin. These include retaining excellent Gram-negative bacilli activity, with improved Gram-positive activity. New FQ-like levofloxacin possesses greater bioavailabilty and a longer serum half-life compared with ciprofloxacin, allowing for once-daily dosing, which may improve patient adherence. The high bioavailability of levofloxacin allows for rapid step-down from intravenous administration to oral therapy, minimizing unnecessary hospitalization, which may decrease costs and improve patient quality of life. Levofloxacin has been evaluated for the treatment of community-acquired pneumonia (CAP) in numerous randomized clinical trials. Most published studies have used the 500 mg dose, although more recent studies have investigated the 750 mg dose once daily. These trials demonstrate that levofloxacin is effective and safe for the treatment of CAP, displaying relatively mild adverse effects that are more or less comparable with ciprofloxacin. Levofloxacin has much to offer in terms of bacterial eradication, including for resistant respiratory pathogens. However, ciprofloxacin-resistant organisms are becoming more prevalent so prudence must be exercised when prescribing this agent.

Semisynthetic Penicillin 6-[d(—)-α-Carboxy-3-Thienylacetamido] Penicillanic Acid Active Against Pseudomonas In Vitro

PubMed Central

Neu, H. C.; Winshell, E. B.

1971-01-01

The activity of 6-[d(—)-α-carboxy-3-thienylacetamido] penicillanic acid, BRL2288, was determined against Pseudomonas aeruginosa and various gram-negative bacilli. The majority of Pseudomonas strains (89%) were inhibited by 100 μg of the antibiotic per ml. BRL2288 is twofold more active than carbenicillin against Pseudomonas at 100 μg/ml or less. Among Enterobacteriaceae tested, 87% Enterobacter and 87% of Proteus mirabilis strains were inhibited by 25 μg/ml or less. Indole-positive Proteus were inhibited by 10 μg/ml or less. Fifty-five per cent of ampicillin-resistant Escherichia coli were inhibited by 100 μg/ml. Klebsiella were uniformly resistant. BRL2288 is not hydrolyzed by most resistant Pseudomonas, but it is destroyed by the β-lactamases of E. coli and P. mirabilis. The antibiotic shows synergy with gentamicin but not with penicillinase-resistant penicillins such as cloxacillin. Activity of BRL2288 against gram-positive organisms is two- to eightfold less than that of ampicillin or benzylpenicillin G. PMID:4993233

PIRATE project: point-of-care, informatics-based randomised controlled trial for decreasing overuse of antibiotic therapy in Gram-negative bacteraemia.

PubMed

Huttner, Angela; Albrich, Werner C; Bochud, Pierre-Yves; Gayet-Ageron, Angèle; Rossel, Anne; Dach, Elodie von; Harbarth, Stephan; Kaiser, Laurent

2017-07-13

Antibiotic overuse drives antibiotic resistance. The optimal duration of antibiotic therapy for Gram-negative bacteraemia (GNB), a common community and hospital-associated infection, remains unknown and unstudied via randomised controlled trials (RCTs). This investigator-initiated, multicentre, non-inferiority, informatics-based point-of-care RCT will randomly assign adult hospitalised patients receiving microbiologically efficacious antibiotic(s) for GNB to (1) 14 days of antibiotic therapy, (2) 7 days of therapy or (3) an individualised duration determined by clinical response and 75% reduction in peak C reactive protein (CRP) values. The randomisation will occur in equal proportions (1:1:1) on day 5 (±1) of efficacious antibiotic therapy as determined by antibiogram; patients, their physicians and study investigators will be blind to treatment duration allocation until the day of antibiotic discontinuation. Immunosuppressed patients and those with GNB due to complicated infections (endocarditis, osteomyelitis, etc) and/or non-fermenting bacilli ( Acinetobacter spp, Burkholderia spp, Pseudomonas spp) Brucella spp, Fusobacterium spp or polymicrobial growth with Gram-positive organisms will be ineligible. The primary outcome is incidence of clinical failure at day 30; secondary outcomes include clinical failure, all-cause mortality and incidence of Clostridiumdifficile infection in the 90-day study period. An interim safety analysis will be performed after the first 150 patients have been followed for ≤30 days. Given a chosen margin of 10%, the required sample size to determine non-inferiority is roughly 500 patients. Analyses will be performed on both intention-to-treat and per-protocol populations. Ethics approval was obtained from the cantonal ethics committees of all three participating sites. Results of the main trial and each of the secondary endpoints will be submitted for publication in a peer-reviewed journal. This trial is registered at www.clinicaltrials.gov (NCT03101072; pre-results). © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Risk factors of all-cause in-hospital mortality among Korean elderly bacteremic urinary tract infection (UTI) patients.

PubMed

Chin, Bum Sik; Kim, Myung Soo; Han, Sang Hoon; Shin, So Youn; Choi, Hee Kyung; Chae, Yun Tae; Jin, Sung Joon; Baek, Ji-Hyeon; Choi, Jun Yong; Song, Young Goo; Kim, Chang Oh; Kim, June Myung

2011-01-01

Urinary tract infection (UTI) is the most frequent cause of bacteremia/sepsis in elderly people and increasing antimicrobial resistance in uropathogens has been observed. To describe the characteristics of bacteremic UTI in elderly patients and to identify the independent risk factors of all-cause in-hospital mortality, a retrospective cohort study of bacteremic UTI patients of age over 65 was performed at a single 2000-bed tertiary hospital. Bacteremic UTI was defined as the isolation of the same organism from both urine and blood within 48 h. Eighty-six elderly bacteremic UTI patients were enrolled. Community-acquired infection was the case for most patients (79.1%), and Escherichia coli accounted for 88.6% (70/79) among Gram-negative organisms. Non-E. coli Gram-negative organisms were more frequent in hospital-acquired cases and male patients while chronic urinary catheter insertion was related with Gram-positive urosepsis. The antibiotic susceptibility among Gram-negative organisms was not different depending on the source of bacteremic UTI, while non-E. coli Gram-negative organisms were less frequently susceptible for cefotaxime, cefoperazone/sulbactam, and aztreonam. All-cause in-hospital mortality was 11.6%, and functional dependency (adjusted hazard ratio=HR=10.9, 95% confidence interval=95%CI=2.2-54.6) and low serum albumin (adjusted HR=27.0, 95%CI=2.0-361.2) were independently related with increased all-cause in-hospital mortality. Crown Copyright © 2010. Published by Elsevier Ireland Ltd. All rights reserved.

Osteomyelitis associated with Nocardiopsis composta in a dog.

PubMed

Salas, Elisa N; Royal, Debra; Kurz, Lance; Loy, J Dustin

2015-05-01

We report the first detection of Nocardiopsis composta in association with osteomyelitis in a young male miniature Australian shepherd dog. Findings included suppurative osteomyelitis containing intralesional Fite's acid fast bacilli, aerobic culture of branching Gram-positive rods, and positive identification via phenotypic analysis and 16S rDNA sequencing.

A brief history of infection control - past and present.

PubMed

Forder, A A

2007-11-01

The scientific study of hospital or nosocomial cross-infection began during the first half of the 18th century, and from that time until the start of the 'Bacteriological Era' many of the most notable contributions originated in Scotland. However it was only 100 years later in 1858 that Florence Nightingale promoted the case for hospital reform. The real understanding of hospital infection followed upon the discoveries of Pasteur, Koch and Lister and the beginning of the 'Bacteriological Era'. The close of the 19th century saw the triumphs of hospital reform and asepsis and seemed to herald the final victory over hospital cross-infection. However, the victory was short-lived. It was soon realised that infections occurred not only in obstetric and surgical patients, but in medical patients as well, and that air could also be a source of infection. Streptococcal, staphylococcal and then Gram-negative bacilli as a cause of hospital infection became a focus of attention, as did antibiotic-resistant organisms. This paper looks briefly at the establishment of the control of infection doctor, infection control committee and infection control nurse as well as summarising the changes, problems and advances in infection control up to the present time.

Oropharyngeal dysphagia pathophysiology, complications and science-based interventions.

PubMed

Altman, Kenneth W

2012-01-01

The etiology of oropharyngeal dysphagia can be broad, and includes aging with atrophy, debilitation, stroke, neurodegenerative and muscular diseases, tumor and postsurgical deformity, as well as effects due to medications and drying of the mucosal membranes. Pathophysiology depends on the multiple causative factors, including the cortex and neural connections to generate the swallow, as well as the oropharyngeal musculature. While chronic debilitation and age may result in nutritional deficiency and poor hydration, the other causes generally present with aspiration risk more acutely. Bacteriologically, aspiration pneumonia is usually polymicrobial with a predominance of Gram-negative enteric bacilli. However, there is emerging evidence to suggest that odontogenic sources may complicate the severity of bacterial load. The principles behind science-based interventions are primarily aspiration assessment with bedside evaluation, and ultimately modified barium swallow (videofluoroscopy) or functional endoscopic evaluation of swallowing (with or without sensory testing). Each has its advantages and logistical concerns. Intervention and rehabilitation is unique to the patient's needs, but may include reconditioning and therapy with a speech and language pathologist, and surgical options. The emerging roles of neuroplasticity and external neuromuscular stimulation are also discussed. Copyright © 2012 S. Karger AG, Basel.

Identification badges: a potential fomite?

PubMed

Ota, Kaede; Profiti, Raffaela; Smaill, Fiona; Matlow, Anne G; Smieja, Marek

2007-01-01

Staff identification badges are mandatory in all hospitals. The purpose of this study was to assess microbial contamination of identification badges at a Canadian tertiary centre. Risk factors for badge contamination were also investigated. Badges were cultured from 118 subjects including secretaries, physicians, nurses, and allied health workers. Subjects also completed a demographic questionnaire. Badge contamination was analyzed according to profession, workplace, duration of badge use, presence of a plastic cover, how the badge was worn, and cleaning frequency. 13.6% of the badges were contaminated with significant pathogens. S. aureus was isolated in 6.8% of the badges, gram-negative bacilli in 5.9%. Contamination was highest in nurses (21.4% versus 9.4-14.3% in other professions) and in the ICU (22.6% versus 8.3%-14.3% at other locations). Neither association was statistically significant. Covered and non-covered badges had similar contamination rates (12% and 17.1%) as did badges worn around the neck compared with those worn clipped to clothing (13.0% versus 14.6%). Contamination of recently cleaned badges was not statistically different from those that had not. Identification badges do not appear to be a major reservoir for pathogenic organisms. Badges can, however, harbour disease-causing organisms and should be cleaned regularly.

Current Microbiology of Surgical Site Infections in Patients with Cancer: A Retrospective Review.

PubMed

Rolston, Kenneth V I; Nesher, Lior; Tarrand, Jeffrey T

2014-12-01

Patients with solid tumors frequently undergo surgical procedures and develop procedure-related infections. We sought to describe the current microbiologic spectrum of infections at various sites following common surgical procedures. This was a retrospective review of microbiologic data between January 2011 and February 2012. The sites studied were those associated with breast cancer surgery, thoracotomy, craniotomy, percutaneous endoscopic gastrostomy (PEG) tube insertion, and abdominal/pelvic surgery. Only patients with solid tumors were included. A total of 368 surgical site infections (SSIs) were identified (68 breast cancer related; 91 thoracotomy related; 45 craniotomy related; 75 PEG-tube insertion related; and 89 abdominal/pelvic surgery related). Of these, 58% were monomicrobial and 42% were polymicrobial. Overall, 85% of the 215 monomicrobial infections were caused by Gram-positive organisms and 13% by Gram-negative bacilli (GNB). Staphylococcus aureus was the predominant pathogen in monomicrobial infections (150 of 215, 70%). Sixty (40%) of these staphylococcal isolates were methicillin resistant (MRSA), and 65% had a vancomycin minimal inhibitory concentration (MIC) ≥1.0 µg/ml. Pseudomonas aeruginosa was the predominant GNB pathogen (19 of 27, 70%). Staphylococci were also the predominant pathogens in polymicrobial infections, while P. aeruginosa and Escherichia coli were the predominant GNB. Overall, 35% of isolates from polymicrobial infections were GNB. Cephalosporins (e.g., cefazolin) or amoxicillin/clavulanate was used most often for surgical prophylaxis, and 47% of organisms from monomicrobial infections (MRSA, P. aeruginosa) were resistant to them. A similar resistance pattern was observed in polymicrobial infections. Staphylococcus species were isolated most often from the sites studied. Polymicrobial infections (42%) and GNB monomicrobial infections (13%) were relatively frequent causes of SSIs. Many of these infections were caused by organisms that are resistant to agents commonly used for surgical prophylaxis. Additionally, 65% of staphylococcal isolates had a vancomycin MIC ≥1.0 µg/ml, suggesting the need for alternative therapeutic agents.

Compliance of clinical microbiology laboratories in the United States with current recommendations for processing respiratory tract specimens from patients with cystic fibrosis.

PubMed

Zhou, Juyan; Garber, Elizabeth; Desai, Manisha; Saiman, Lisa

2006-04-01

Respiratory tract specimens from patients with cystic fibrosis (CF) require unique processing by clinical microbiology laboratories to ensure detection of all potential pathogens. The present study sought to determine the compliance of microbiology laboratories in the United States with recently published recommendations for CF respiratory specimens. Microbiology laboratory protocols from 150 of 190 (79%) CF care sites were reviewed. Most described the use of selective media for Burkholderia cepacia complex (99%), Staphylococcus aureus (82%), and Haemophilus influenzae (89%) and identified the species of all gram-negative bacilli (87%). Only 52% delineated the use of agar diffusion assays for susceptibility testing of Pseudomonas aeruginosa. Standardizing laboratory practices will improve treatment, infection control, and our understanding of the changing epidemiology of CF microbiology.

[A parotitis as primary infection of Lemierre's syndrome].

PubMed

Valleix, B; Floccard, B; Hautin, E; Faure, F; Allaouchiche, B

2011-09-01

Lemierre's syndrome is a classical presentation of human necrobacillosis. It is characterized by a primary infection in the face including a septic thrombophlebitis of the internal jugular vein and disseminated metastatic abcesses. Fusobacterium necrophorum is the main pathogen found in that syndrome. The diagnosis is based on clinical features, then on the microbiology with positive anaerobic blood cultures as key role and finally on the computed tomography. Most of the time a well-chosen antibiotic treatment against anaerobic pathogens and Gram negative bacilli is efficient but surgery can be useful. We report a case of a 73 years old man, which seems to be unique because it is the first case reported of a Lemierre's syndrome characterized by a parotitis infected by F. necrophorum. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

KPC-4 Is Encoded within a Truncated Tn4401 in an IncL/M Plasmid, pNE1280, Isolated from Enterobacter cloacae and Serratia marcescens

PubMed Central

Bryant, Kendall A.; Van Schooneveld, Trevor C.; Thapa, Ishwor; Bastola, Dhundy; Williams, Laurina O.; Safranek, Thomas J.; Hinrichs, Steven H.; Rupp, Mark E.

2013-01-01

We describe the transfer of blaKPC-4 from Enterobacter cloacae to Serratia marcescens in a single patient. DNA sequencing revealed that KPC-4 was encoded on an IncL/M plasmid, pNE1280, closely related to pCTX-M360. Further analysis found that KPC-4 was encoded within a novel Tn4401 element (Tn4401f) containing a truncated tnpA and lacking tnpR, ISKpn7 left, and Tn4401 IRL-1, which are conserved in other Tn4401 transposons. This study highlights the continued evolution of Tn4401 transposons and movement to multiple plasmid backbones that results in acquisition by multiple species of Gram-negative bacilli. PMID:23070154

Bloodstream infections at a tertiary referral hospital in Yangon, Myanmar.

PubMed

Myat, Tin Ohn; Prasad, Namrata; Thinn, Kyi Kyi; Win, Kyu Kyu; Htike, Wah Win; Zin, Khwar Nyo; Murdoch, David R; Crump, John A

2014-11-01

Data regarding characteristics of bloodstream infections in Myanmar are limited. Blood culture results from all outpatients and inpatients were extracted from records of the Clinical Microbiology Laboratory, Yangon General Hospital, for the period 2005 through 2013. Of 3865 blood cultures performed, 449 (11.6%) were positive for a pathogenic organism. Gram-negative bacteria was the most common organism group, accounting for 246 (55.5%) of 449 isolations. Staphylococcus aureus was the most common isolate, detected in 171 (38.1%) of 449 blood cultures. From 2005-2008 to 2009-2013 the proportion of all pathogenic isolates that were Gram-positive declined from 52.8% (167/316) to 20.3% (27/133) (p<0.001), whereas the proportion of Gram-negative bacteria rose from 45.6% (144/316) to 78.9% (105/133) (p<0.001), with non-fermentative bacilli accounting for much of this increase. Antimicrobial susceptibility testing demonstrated a high prevalence resistance of S. aureus to first-line antimicrobials such as erythromycin, penicillin and oxacillin. More than half of tested Escherichia coli and Citrobacter species showed resistance to amoxicillin-clavulanic acid, ceftriaxone or gentamicin. Bloodstream infections are common among patients receiving blood culture at a tertiary hospital in Yangon, Myanmar. Our findings suggest that antimicrobial resistance among invasive bacteria is common, similar to patterns described elsewhere in the region, and highlight the need for locally adapted antimicrobial guidelines for sepsis management. © The Author 2014. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Drosophila immunity: analysis of PGRP-SB1 expression, enzymatic activity and function.

PubMed

Zaidman-Rémy, Anna; Poidevin, Mickael; Hervé, Mireille; Welchman, David P; Paredes, Juan C; Fahlander, Carina; Steiner, Hakan; Mengin-Lecreulx, Dominique; Lemaitre, Bruno

2011-02-18

Peptidoglycan is an essential and specific component of the bacterial cell wall and therefore is an ideal recognition signature for the immune system. Peptidoglycan recognition proteins (PGRPs) are conserved from insects to mammals and able to bind PGN (non-catalytic PGRPs) and, in some cases, to efficiently degrade it (catalytic PGRPs). In Drosophila, several non-catalytic PGRPs function as selective peptidoglycan receptors upstream of the Toll and Imd pathways, the two major signalling cascades regulating the systemic production of antimicrobial peptides. Recognition PGRPs specifically activate the Toll pathway in response to Lys-type peptidoglycan found in most Gram-positive bacteria and the Imd pathway in response to DAP-type peptidoglycan encountered in Gram-positive bacilli-type bacteria and in Gram-negative bacteria. Catalytic PGRPs on the other hand can potentially reduce the level of immune activation by scavenging peptidoglycan. In accordance with this, PGRP-LB and PGRP-SC1A/B/2 have been shown to act as negative regulators of the Imd pathway. In this study, we report a biochemical and genetic analysis of PGRP-SB1, a catalytic PGRP. Our data show that PGRP-SB1 is abundantly secreted into the hemolymph following Imd pathway activation in the fat body, and exhibits an enzymatic activity towards DAP-type polymeric peptidoglycan. We have generated a PGRP-SB1/2 null mutant by homologous recombination, but its thorough phenotypic analysis did not reveal any immune function, suggesting a subtle role or redundancy of PGRP-SB1/2 with other molecules. Possible immune functions of PGRP-SB1 are discussed.

[Antimicrobial therapy in severe infections with multidrug-resistant Gram-negative bacterias].

PubMed

Duszyńska, Wiesława

2010-01-01

Multidrug-resistant Gram-negative bacteria pose a serious and rapidly emerging threat to patients in healthcare settings, and are especially prevalent and problematic in intensive therapy units. Recently, the emergence of pandrug-resistance in Gram-negative bacteria poses additional concerns. This review examines the clinical impact and epidemiology of multidrug-resistant Gram-negative bacteria as a cause of increased morbidity and mortality among ITU patients. Beta-lactamases, cephalosporinases and carbapenemases play the most important role in resistance to antibiotics. Despite the tendency to increased resistance, carbapenems administered by continuous infusion remain the most effective drugs in severe sepsis. Drug concentration monitoring, albeit rarely used in practice, is necessary to ensure an effective therapeutic effect.

Pathogens causing blood stream infections and their drug susceptibility profile in immunocompromised patients.

PubMed

Fayyaz, Muhammad; Mirza, Irfan Ali; Ikram, Aamer; Hussain, Aamir; Ghafoor, Tahir; Shujat, Umer

2013-12-01

To determine the types of pathogens causing blood stream infections and their drug susceptibility profile in immunocompromised patients. Cross-sectional, observational study. Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi, from January to September 2012. Blood culture bottles received from immunocompromised patients were dealt by two methods, brain heart infusion (BHI) broth based manual method and automated BACTEC system. The samples yielding positive growth from either of two methods were further analyzed. The identification of isolates was done with the help of biochemical reactions and rapid tests. Antimicrobial susceptibility of the isolates was carried out as per recommendations of Clinical and Laboratory Standards Institute (CLSI). Out of the 938 blood culture specimens received from immunocompromised patients, 188 (20%) yielded positive growth. Out of these, 89 (47.3%) isolates were Gram positive and Gram negative each, while 10 (5.3%) isolates were fungi (Candida spp.). In case of Gram positive isolates, 75 (84.3%) were Staphylococcus spp. and 51 (67%) were Methicillin resistant. Amongst Gram negative group 49 (55.1%) isolates were of enterobacteriaceae family, while 40 (44.9%) were non-lactose fermenters (NLF). In vitro antimicrobial susceptibility of Staphylococci revealed 100% susceptibility to vancomycin and linezolid. The enterobacteriaceae isolates had better susceptibility against amikacin 85.7% compared to tigecycline 61.2% and imipenem 59.2%. For NLF, the in vitro efficacy of aminoglycosides was 72.5%. The frequency of Gram positive and Gram negative organisms causing blood stream infections in immunocompromised patients was equal. Vancomycin in case of Gram positive and amikacin for Gram negative organisms revealed better in vitro efficacy as compared to other antibiotics.

The impact of metabolic state on Cd adsorption onto bacterial cells

USGS Publications Warehouse

Johnson, K.J.; Ams, D.A.; Wedel, A.N.; Szymanowski, J.E.S.; Weber, D.L.; Schneegurt, M.A.; Fein, J.B.

2007-01-01

This study examines the effect of bacterial metabolism on the adsorption of Cd onto Gram-positive and Gram-negative bacterial cells. Metabolically active Gram-positive cells adsorbed significantly less Cd than non-metabolizing cells. Gram-negative cells, however, showed no systematic difference in Cd adsorption between metabolizing and non-metabolizing cells. The effect of metabolism on Cd adsorption to Gram-positive cells was likely due to an influx of protons in and around the cell wall from the metabolic proton motive force, promoting competition between Cd and protons for adsorption sites on the cell wall. The relative lack of a metabolic effect on Cd adsorption onto Gram-negative compared to Gram-positive cells suggests that Cd binding in Gram-negative cells is focused in a region of the cell wall that is not reached, or is unaffected by this proton flux. Thermodynamic modeling was used to estimate that proton pumping causes the pH in the cell wall of metabolizing Gram-positive bacteria to decrease from the bulk solution value of 7.0 to approximately 5.7. ?? 2007 The Authors.

Clinical manifestations and management of left ventricular assist device-associated infections.

PubMed

Nienaber, Juhsien Jodi C; Kusne, Shimon; Riaz, Talha; Walker, Randall C; Baddour, Larry M; Wright, Alan J; Park, Soon J; Vikram, Holenarasipur R; Keating, Michael R; Arabia, Francisco A; Lahr, Brian D; Sohail, M Rizwan

2013-11-01

Infection is a serious complication of left ventricular assist device (LVAD) therapy. Published data regarding LVAD-associated infections (LVADIs) are limited by single-center experiences and use of nonstandardized definitions. We retrospectively reviewed 247 patients who underwent continuous-flow LVAD implantation from January 2005 to December 2011 at Mayo Clinic campuses in Minnesota, Arizona, and Florida. LVADIs were defined using the International Society for Heart and Lung Transplantation criteria. We identified 101 episodes of LVADI in 78 patients (32%) from this cohort. Mean age (± standard deviation [SD]) was 57±15 years. The majority (94%) underwent Heartmate II implantation, with 62% LVADs placed as destination therapy. The most common type of LVADIs were driveline infections (47%), followed by bloodstream infections (24% VAD related, and 22% non-VAD related). The most common causative pathogens included gram-positive cocci (45%), predominantly staphylococci, and nosocomial gram-negative bacilli (27%). Almost half (42%) of the patients were managed by chronic suppressive antimicrobial therapy. While 14% of the patients had intraoperative debridement, only 3 underwent complete LVAD removal. The average duration (±SD) of LVAD support was 1.5±1.0 years. At year 2 of follow-up, the cumulative incidence of all-cause mortality was estimated to be 43%. Clinical manifestations of LVADI vary on the basis of the type of infection and the causative pathogen. Mortality remained high despite combined medical and surgical intervention and chronic suppressive antimicrobial therapy. Based on clinical experiences, a management algorithm for LVADI is proposed to assist in the decision-making process.

Treatment of Gram-negative bacterial infections by potentiation of antibiotics.

PubMed

Zabawa, Thomas P; Pucci, Michael J; Parr, Thomas R; Lister, Troy

2016-10-01

Infections caused by antibiotic-resistant pathogens, particularly Gram-negative bacteria, represent significant treatment challenges for physicians resulting in high rates of morbidity and mortality. The outer membrane of Gram-negative bacteria acts as a permeability barrier to many compounds that would otherwise be effective antibacterial agents, including those effective against Gram-positive pathogens. Potentiator molecules disrupt this barrier allowing entry of otherwise impermeant molecules, thus providing a strategy to render multi-drug resistant pathogens susceptible to a broader range of antibiotics. Potentiator molecules are cationic and the mechanism of disruption involves interaction with the negatively charged outer membrane. This physical attribute, along with an often high degree of lipophilicity typically endears these molecules with unacceptable toxicity. Presented herein are examples of advanced potentiator molecules being evaluated for use in combination therapy for the treatment of resistant Gram-negative infections. Copyright © 2016 Elsevier Ltd. All rights reserved.

Incidence of carbapenem-resistant gram negatives in Italian transplant recipients: a nationwide surveillance study.

PubMed

Lanini, Simone; Costa, Alessandro Nanni; Puro, Vincenzo; Procaccio, Francesco; Grossi, Paolo Antonio; Vespasiano, Francesca; Ricci, Andrea; Vesconi, Sergio; Ison, Michael G; Carmeli, Yehuda; Ippolito, Giuseppe

2015-01-01

Bacterial infections remain a challenge to solid organ transplantation. Due to the alarming spread of carbapenem-resistant gram negative bacteria, these organisms have been frequently recognized as cause of severe infections in solid organ transplant recipients. Between 15 May and 30 September 2012 we enrolled 887 solid organ transplant recipients in Italy with the aim to describe the epidemiology of gram negative bacteria spreading, to explore potential risk factors and to assess the effect of early isolation of gram negative bacteria on recipients' mortality during the first 90 days after transplantation. During the study period 185 clinical isolates of gram negative bacteria were reported, for an incidence of 2.39 per 1000 recipient-days. Positive cultures for gram negative bacteria occurred early after transplantation (median time 26 days; incidence rate 4.33, 1.67 and 1.14 per 1,000 recipient-days in the first, second and third month after SOT, respectively). Forty-nine of these clinical isolates were due to carbapenem-resistant gram negative bacteria (26.5%; incidence 0.63 per 1000 recipient-days). Carbapenems resistance was particularly frequent among Klebsiella spp. isolates (49.1%). Recipients with longer hospital stay and those who received either heart or lung graft were at the highest risk of testing positive for any gram negative bacteria. Moreover recipients with longer hospital stay, lung recipients and those admitted to hospital for more than 48h before transplantation had the highest probability to have culture(s) positive for carbapenem-resistant gram negative bacteria. Forty-four organ recipients died (0.57 per 1000 recipient-days) during the study period. Recipients with at least one positive culture for carbapenem-resistant gram negative bacteria had a 10.23-fold higher mortality rate than those who did not. The isolation of gram-negative bacteria is most frequent among recipient with hospital stays >48 hours prior to transplant and in those receiving either heart or lung transplants. Carbapenem-resistant gram negative isolates are associated with significant mortality.

Prevalence of communicable disease among restaurant workers along a highway in Assam, India.

PubMed

Biswas, D; Hazarika, N C; Hazarika, D; Mahanta, J

1999-09-01

105 workers of 29 roadside restaurants along a highway in Assam were interviewed for their habits, educational status and were examined for their hygienic status and presence of communicable diseases (STDs,TB etc). Most of them were young, males with 40% literacy; more than half were unmarried. About 30% of them were alcoholic and smokers and 2.9% were addicted to cannabis. The hygienic conditions of the workers were poor. More than one third had sexual contact with multiple sex partners or commercial sex workers and 2% were engaged in homosexual activity. Most of them did not use condom. 25.7% of them had genital lesions suggestive of sexually transmitted disease, 11.8% showed gram negative diplococci in urethral smears and 5.1% were VDRL reactive. Skin infections followed by gastrointestinal disorders and respiratory tract infections were other prevalent problems. 70.6% were positive for intestinal parasites and 22.2% were sputum positive for acid fast bacilli. However all the collected blood samples were negative for HIV.

Characterization of wound infections among patients injured during the 2011 Libyan conflict.

PubMed

Dau, A A; Tloba, S; Daw, M A

2013-04-01

Few studies have analysed the bacterial pathogenesis of infections associated with war-wound in the Eastern Mediterranean region. We analysed surgical wound infections of 1200 patients injured during the Libyan conflict in 2011 and admitted to the emergency services at Tripoli medical centre. Culture swabs or surgical wound debridement samples were collected and cultures were identified and tested for antimicrobial resistance. Of the 1200 patients studied, 498 (42%) were infected with at least 1 pathogen and 57 with >2 pathogens. The most common species were Acinetobacter spp. (isolated from 144 patients), coagulase-negative staphylococci (122), Escherichia coli (107), Pseudomnonas aeruginosa (92) and Klebsiella spp. (86). A high level of resistance to the antibiotics tested was found, especially among Acinetobacter spp. Multi-drug-resistant Gram-negative bacilli were an important complicating factor in wound infections associated with war injuries among injured patients in Libya. Effective policies are needed to control and treat such infections particularly in trauma and emergency services.

Cutaneous anthrax in an unusual location: case report.

PubMed

Sari, Tugba; Koruk, Suda Tekin

2015-12-01

Cutaneous anthrax is well known, unlike anthrax of the lumbar region, which is not reported elsewhere. We present a case of anthrax of the lumbar region in a 50-year-old man. The infection was characterised by a wide, black eschar and oedema on an erythematous ground. After isolation of the Gram-positive bacilli from the skin lesions, prompt antibiotic treatment (intravenous sulbactam-ampicillin 1.5 g every six hours) was initiated. Following eradication of the bacilli after 14 days of antibiotic treatment, a split-thickness skin graft was applied. A diagnosis of anthrax depends on clinical suspicion. Early diagnosis, antibiotic and surgical treatment can facilitate the treatment and prevent development of complications.

Impact of ertapenem on antimicrobial resistance in a sentinel group of Gram-negative bacilli: a 6 year antimicrobial resistance surveillance study.

PubMed

Rodriguez-Osorio, Carlos A; Sanchez-Martinez, Cesar O; Araujo-Melendez, Javier; Criollo, Elia; Macias-Hernandez, Alejandro E; Ponce-de-Leon, Alfredo; Ponce-de-Leon, Sergio; Sifuentes-Osornio, Jose

2015-03-01

To determine the association between ertapenem and resistance of Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Acinetobacter baumannii-calcoaceticus complex to different antimicrobials while adjusting for relevant hospital factors. This was a retrospective time-series study conducted at a tertiary care centre from September 2002 to August 2008. The specific impact of ertapenem on the resistance of these Gram-negative bacilli (GNB) was assessed by multiple linear regression analysis, adjusting for the average length of stay, rate of hospital-acquired infections and use of 10 other antimicrobials, including type 2 carbapenems. Unadjusted analyses revealed significant increases over the duration of the study in the number of GNB resistant to meropenem/imipenem among 1000 isolates each of E. coli (0.46 ± 0.22, P < 0.05), P. aeruginosa (6.26 ± 2.26, P < 0.05), K. pneumoniae (8.06 ± 1.50, P < 0.0005) and A. baumannii-calcoaceticus complex (25.39 ± 6.81, P < 0.0005). Increased resistance to cefepime (and other extended-spectrum cephalosporins) was observed in E. coli (9.55 ± 1.45, P < 0.0005) and K. pneumoniae (15.21 ± 2.42, P < 0.0005). A. baumannii-calcoaceticus complex showed increased resistance to all antimicrobials except amikacin. After controlling for confounders, ertapenem was not significantly associated (P > 0.05) with changes in resistance for any pathogen/antimicrobial combination. After controlling for confounders, ertapenem was not associated with changes in resistance in a group of sentinel GNB, although significant variations in resistance to different antimicrobials were observed in the unadjusted analyses. These results emphasize the importance of implementation of local resistance surveillance platforms and stewardship programmes to combat the global emergence and spread of antimicrobial resistance. © The Author 2014. 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.

When it comes to antibiotics, bacteria show some NO-how.

PubMed

Patel, Bhumit A; Crane, Brian R

2010-10-01

Homologs to mammalian nitric oxide synthases are found in many mostly Gram-positive bacteria. In some genera such as bacilli, and staphylococci, these enzymes produce protects against oxidative damage, this effect has now been shown to provide an advantage against antibiotics that kill by increasing cellular levels of reactive oxygen species.

Osteomyelitis associated with Nocardiopsis composta in a dog

PubMed Central

Salas, Elisa N.; Royal, Debra; Kurz, Lance; Loy, J. Dustin

2015-01-01

We report the first detection of Nocardiopsis composta in association with osteomyelitis in a young male miniature Australian shepherd dog. Findings included suppurative osteomyelitis containing intralesional Fite’s acid fast bacilli, aerobic culture of branching Gram-positive rods, and positive identification via phenotypic analysis and 16S rDNA sequencing. PMID:25969577

Resistance surveillance program report for selected European nations (2011).

PubMed

Jones, Ronald N; Flonta, Mirela; Gurler, Nazahat; Cepparulo, Mario; Mendes, Rodrigo E; Castanheira, Mariana

2014-04-01

In the European component of the Regional Resistance Surveillance study for 2011, a total of 21 countries were monitored for antimicrobial resistance patterns including Belgium, Bulgaria (BU), Croatia, Czech Republic, France (F), Germany (GE), Greece (GR), Ireland (IR), Israel (IS), Italy (IT), Poland (PO), Portugal (PT), Romania (RO), Russia (RU), Slovakia (SK), Slovenia, Spain, Sweden (SW), Turkey (T), Ukraine, and United Kingdom. Results from testing 12,572 strains (100 [BU] to 1535 [F] per nation) were interpreted by contemporary published breakpoints. Samples from 47 hospitals were reference tested against agents such as amikacin (AMK), cefoperazone/sulbactam (C/S), colistin (COL), levofloxacin, linezolid (LZD), tigecycline (TIG), vancomycin (VAN), and 21 others. Among Staphylococcus aureus, LZD (MIC90, 2 μg/mL), TIG (MIC90, 0.12 μg/mL), and VAN (MIC90, 1 μg/mL) exhibited complete coverage and methicillin resistance rates among nations (average, 31%) ranged from 0.9% (SW) to 60.0-60.2% (PT, SK). Seven LZD-resistant coagulase-negative staphylococci (only 1.1% resistance overall) were noted in 5 nations, and a Staphylococcus simulans strain (MIC, 8 μg/mL from RO) had L3 mutations (N130D, G152A, F147S, A157R); also 6 LZD-resistant enterococci were detected in 3 countries (GE, IR, T). VAN-resistant enterococci (10% overall; 84% VanA) were found in 14 countries, highest in GE and IR (23.0%). The ESBL phenotype rate for Escherichia coli was 20.1% (range, 0.9% [SW] to 70.0-89.7% [BU, RU]), best inhibited by COL (100.0% S), TIG (100.0%), AMK (83.3-94.1%), C/S (81.0%), and carbapenems (>99.0%; resistant strains in IS and T). Klebsiella spp. had greater ESBL rates (45.7% overall, range 2.5-100.0%), as well as carbapenem resistance (8.3% overall, greatest in BU, GR, IS, IT, PO, RO, RU, T). Non-fermentative Gram-negative bacilli (Pseudomonas aeruginosa, Acinetobacter [ACB]) were generally less susceptible, except against COL (99.2-99.6% S) and TIG (95.0% inhibited at ≤2 μg/mL; ACB only). The following carbapenemases were detected: VIM-1 (2 countries); IMP-1 (1 from T); KPC-2 or -3 (2 countries); VIM-4 (1 from PO), NDM-1 (2 in RO; 2 centers); and OXA-48 or -162 (5 from T; 2 centers). European surveillance sampling demonstrates a wide array of resistant isolates, less prevalent among Gram-positive cocci that remain inhibited by several available agents. However, beta-lactamase-mediated mechanisms have spread widely among Gram-negative bacilli, especially across the Eastern and Southern European nations, severely limiting infection chemotherapy and necessitating escalated antimicrobial stewardship. © 2014.

Emerging Trends of Bloodstream Infections: A Six-Year Study at a Paediatric Tertiary Care Hospital in Kabul.

PubMed

Tariq, Tariq Mahmud; Rasool, Esmatullah

2016-11-01

To determine the frequency of pathogens causing bloodstream infections and evaluate their trends and antibiogram patterns among in-patients in a paediatric tertiary care centre. Descriptive study. French Medical Institute for Mothers and Children (FMIC), Kabul, Afghanistan in two phases, from January 2010 to December 2015. Results of blood cultures from suspected cases of sepsis admitted in the FMIC, from January 2010 to December 2012 (Period-1), and from January 2013 to December 2015 (Period-2) were completed. Standard microbiological methods were followed for blood culture and antibiotic sensitivity testing. Out of total 1,040 cases of culture proven sepsis, 528 (50.77%) Gram-negative bacilli (GNB), 474 (45.58%) Gram-positive cocci (GPC), and 38 (3.65%) Candida species were isolated during the entire study period. Out of 528 GNB isolates, 373 (70.64%) belonged to the Enterobacteriaceae and 155 (29.36%) were non-fermenters. Among Enterobacteriaceae, 168 (31.82%) were Klebsiella species (K. pneumoniae=124, K. oxytoca=44), 70 (13.26%) were Enterobacter species (E. cloacae=52, E. aerogenes=18), 65 (12.31%) were E. coli, 37 (7.01%) were Serratia marcescens and 31 (5.87%) were others. Out of 155 non-fermenters, 88 (16.67%) were Pseudomonas aeruginosa, 39 (7.39%) were Burkholderia cepacia and 18 (3.41%) were Stenotrophomonas maltophilia. There was a drop in the frequency of Enterobacteriaceae from 85% in Period-1 to 58.68% in Period-2. There was an increase in the frequency of nonfermenters from 15% to 41.32%, particularly 18 new cases of sepsis caused by Stenotrophomonas maltophilia during Period-2. Among GPC, there was an overall rise of 16.14% in the prevalence of Staphylococcus epidermidis during Period-2 and a drop of 9.64% in the frequency of Staphylococcus aureus during Period-2. The majority of Gram-negative isolates were multidrug-resistant to commonly used antibiotics. However, most of the isolates were sensitive to amikacin and imipenem (except S. maltophilia). The frequency of those producing ESBL reduced by 11.22% during the Period-2. Among Gram-positive cocci, the pattern of antibiogram did not show a significant change during both periods, and majority remained resistant to commonly used antibiotics. All Staphylococci were sensitive to vancomycin but resistant to penicillin. There was a substantial decline of 18.87% in the frequency of Methicillin-resistant Staphylococci (MRSA/MRSE) during Period-2. Staphylococci and Klebsiellae remain the most important bacteria responsible for bloodstream infections in a tertiary healthcare facility in Kabul. Yet, there has been an increase in the prevalence of Pseudomonas and Burkholderia cepacia. Moreover, Stenotrophomonas maltophilia emerged as a new hospital acquired pathogen. This study could possibly help in suggesting choices eluding the misuse of appropriate antibiotics.

Potentiating the Activity of Nisin against Escherichia coli

PubMed Central

Zhou, Liang; van Heel, Auke J.; Montalban-Lopez, Manuel; Kuipers, Oscar P.

2016-01-01

Lantibiotics are antimicrobial (methyl)lanthionine-containing peptides produced by various Gram-positive bacteria. The model lantibiotic, nisin, binds lipid II in the cell membrane. Additionally, after binding it can insert into the membrane creating a pore. Nisin can efficiently inhibit the growth of Gram-positive bacteria and resistance is rarely observed. However, the activity of lantibiotics is at least 100-fold lower against certain Gram-negative bacteria. This is caused by the fact that Gram-negative bacteria have an outer membrane that hinders the peptides to reach lipid II, which is located in the inner membrane. Improving the activity of lantibiotics against Gram-negative bacteria could be achieved if the outer membrane traversing efficiency is increased. Here, several anti-Gram-negative peptides (e.g., apidaecin 1b, oncocin), or parts thereof, were fused to the C-terminus of either a truncated version of nisin containing the first three/five rings or full length nisin. The activities of these fusion peptides were tested against Gram-negative pathogens. Our results showed that when an eight amino acids (PRPPHPRL) tail from apidaecin 1b was attached to nisin, the activity of nisin against Escherichia coli CECT101 was increased more than two times. This research presents a new and promising method to increase the anti-Gram-negative activity of lantibiotics. PMID:26904542

Genomic and Molecular Characterization of Clinical Isolates of Enterobacteriaceae Harboring mcr-1 in Colombia, 2002 to 2016.

PubMed

Saavedra, Sandra Yamile; Diaz, Lorena; Wiesner, Magdalena; Correa, Adriana; Arévalo, Stefany Alejandra; Reyes, Jinnethe; Hidalgo, Andrea Melissa; de la Cadena, Elsa; Perenguez, Marcela; Montaño, Lucy Angeline; Ardila, Javier; Ríos, Rafael; Ovalle, María Victoria; Díaz, Paula; Porras, Paola; Villegas, Maria V; Arias, Cesar A; Beltrán, Mauricio; Duarte, Carolina

2017-12-01

Polymyxins are last-resort antimicrobial agents used to treat infections caused by carbapenem-resistant Enterobacteriaceae Due to the worldwide dissemination of polymyxin resistance in animal and human isolates, we aimed to characterize polymyxin resistance associated with the presence of mcr-1 in Enterobacteriaceae and nonfermenter Gram-negative bacilli, using isolates collected retrospectively in Colombia from 2002 to 2016. A total of 5,887 Gram-negative clinical isolates were studied, and 513 were found to be resistant to the polymyxins. Susceptibility to colistin was confirmed by broth microdilution for all mcr-1 -positive isolates, and these were further subjected to whole-genome sequencing (WGS). The localization of mcr-1 was confirmed by S1 pulsed-field gel electrophoresis (S1-PFGE) and CeuI-PFGE hybridization. Transferability was evaluated by mating assays. A total of 12 colistin-resistant isolates recovered after 2013 harbored mcr-1 , including 8 Escherichia coli , 3 Salmonella enterica serovar Typhimurium, and 1 Klebsiella pneumoniae isolate . E. coli isolates were unrelated by PFGE and belonged to 7 different sequence types (STs) and phylogroups. S Typhimurium and K. pneumoniae isolates belonged to ST34 and ST307, respectively. The mcr-1 gene was plasmid borne in all isolates but two E. coli isolates which harbored it on the chromosome. Conjugation of mcr-1 was successful in 8 of 10 isolates (8.2 × 10 -5 to 2.07 × 10 -1 cell per recipient). Plasmid sequences showed that the mcr-1 plasmids belonged to four different Inc groups (a new IncP-1 variant and the IncFII, IncHI1, and IncH families). Our results indicate that mcr-1 is circulating in clinical isolates of colistin-resistant Enterobacteriaceae in Colombia and is mainly harbored in transferable plasmids. Copyright © 2017 American Society for Microbiology.

Genomic and Molecular Characterization of Clinical Isolates of Enterobacteriaceae Harboring mcr-1 in Colombia, 2002 to 2016

PubMed Central

Diaz, Lorena; Wiesner, Magdalena; Correa, Adriana; Arévalo, Stefany Alejandra; Reyes, Jinnethe; Hidalgo, Andrea Melissa; de la Cadena, Elsa; Perenguez, Marcela; Montaño, Lucy Angeline; Ardila, Javier; Ríos, Rafael; Ovalle, María Victoria; Díaz, Paula; Porras, Paola; Villegas, Maria V.; Arias, Cesar A.; Beltrán, Mauricio

2017-01-01

ABSTRACT Polymyxins are last-resort antimicrobial agents used to treat infections caused by carbapenem-resistant Enterobacteriaceae. Due to the worldwide dissemination of polymyxin resistance in animal and human isolates, we aimed to characterize polymyxin resistance associated with the presence of mcr-1 in Enterobacteriaceae and nonfermenter Gram-negative bacilli, using isolates collected retrospectively in Colombia from 2002 to 2016. A total of 5,887 Gram-negative clinical isolates were studied, and 513 were found to be resistant to the polymyxins. Susceptibility to colistin was confirmed by broth microdilution for all mcr-1-positive isolates, and these were further subjected to whole-genome sequencing (WGS). The localization of mcr-1 was confirmed by S1 pulsed-field gel electrophoresis (S1-PFGE) and CeuI-PFGE hybridization. Transferability was evaluated by mating assays. A total of 12 colistin-resistant isolates recovered after 2013 harbored mcr-1, including 8 Escherichia coli, 3 Salmonella enterica serovar Typhimurium, and 1 Klebsiella pneumoniae isolate. E. coli isolates were unrelated by PFGE and belonged to 7 different sequence types (STs) and phylogroups. S. Typhimurium and K. pneumoniae isolates belonged to ST34 and ST307, respectively. The mcr-1 gene was plasmid borne in all isolates but two E. coli isolates which harbored it on the chromosome. Conjugation of mcr-1 was successful in 8 of 10 isolates (8.2 × 10−5 to 2.07 × 10−1 cell per recipient). Plasmid sequences showed that the mcr-1 plasmids belonged to four different Inc groups (a new IncP-1 variant and the IncFII, IncHI1, and IncH families). Our results indicate that mcr-1 is circulating in clinical isolates of colistin-resistant Enterobacteriaceae in Colombia and is mainly harbored in transferable plasmids. PMID:28893788

Efficacy of polymyxin B-immobilized fiber hemoperfusion for patients with septic shock caused by Gram-negative bacillus infection.

PubMed

Saito, Nobuyuki; Sugiyama, Kazuhiro; Ohnuma, Testu; Kanemura, Takashi; Nasu, Michitaka; Yoshidomi, Yuya; Tsujimoto, Yuta; Adachi, Hiroshi; Koami, Hiroyuki; Tochiki, Aito; Hori, Kota; Wagatsuma, Yukiko; Matsumoto, Hisashi

2017-01-01

Septic shock-associated mortality in intensive care units (ICUs) remains high, with reported rates ranging 30-50%. In particular, Gram-negative bacilli (GNB), which induce significant inflammation and consequent multiple organ failure, are the etiological bacterial agent in 40% of severe sepsis cases. Hemoperfusion using polymyxin B-immobilized fiber (PMX), which adsorbs endotoxin, is expected to reduce the inflammatory sepsis cascade due to GNB. However, the clinical efficacy of this treatment has not yet been demonstrated. Here, we aimed to verify the efficacy of endotoxin adsorption therapy using PMX through a retrospective analysis of 413 patients who received broad spectrum antimicrobial treatment for GNB-related septic shock between January 2009 and December 2012 in 11 ICUs of Japanese tertiary hospitals. After aligning the patients' treatment time phases, we classified patients in two groups depending on whether PMX hemoperfusion (PMXHP) therapy was administered or not within 24 hours after ICU admission (PMXHP group: n = 134, conventional group: n = 279). The primary study endpoint was the mortality rate at 28 days after ICU admission. The mean age was 72.4 (standard deviation: 12.6) years, and the mean Sequential Organ Failure Assessment score at ICU admission was 9.9 (3.4). The infection sites included intra-abdominal (38.0%), pulmonary (18.9%), and urinary tract (32.2%), and two thirds of all patients had GNB-related bacteremia. Notably, the mortality at 28 days after ICU admission did not differ between the groups (PMXHP: 29.1% vs. conventional: 29.0%, P = 0.98), and PMXHP therapy was not found to improve this outcome in a Cox regression analysis (hazard ratio = 1.16; 95% confidence interval, 0.81-1.64, P = 0.407). We conclude that PMX-based endotoxin adsorption within 24 hours from ICU admission was not associated with mortality among patients with septic shock due to GNB. University Hospital Medical Information Network Clinical Trial Registry (UMIN-CTR ID: UMIN000012748).

Consequences of avoiding β-lactams in patients with β-lactam allergies.

PubMed

Jeffres, Meghan N; Narayanan, Prasanna P; Shuster, Jerrica E; Schramm, Garrett E

2016-04-01

The choice of empiric antibiotics for the treatment of gram-negative bacilli (GNB) bloodstream infections (BSIs) in patients presenting with a β-lactam (BL) allergy is often a difficult decision given that these agents are first-line treatment in many guidelines. We sought to compare rates of clinical failure between patients with a history of BL allergy who received either a BL or a non-β-lactam (NBL). Adult patients with a past medical history of BL allergy and receipt of antibiotics for treatment of a GNB BSI were included from 3 academic medical centers. Treatment groups were classified as BL or NBL groups based on the empiric antibiotics received. Clinical failure was assessed 72 to 96 hours after initiation of empiric antibiotics. Hypersensitivity reactions during receipt of antibiotic therapy for the index BSI were recorded. A total of 552 patients were included for analysis: 433 patients in the BL group and 119 patients in the NBL group. Clinical failure was higher in the NBL group compared with the BL group (38.7% vs 27.4%, P = .030). The most common cause of clinical failure was a temperature of greater than 38.0°C 72 to 96 hours after receipt of empiric antibiotics (NBL group: 22.7% vs BL group: 13.9%, P = .016). Hypersensitivity occurred in 16 (2.9%) of 552 patients. Thirteen (2.5%) of 552 patients experiencing hypersensitivity reactions were exposed to a BL during treatment for GNB BSI. Among patients with a BL allergy, use of BL antibiotics is associated with a lower rate of clinical failure. The low rate of hypersensitivity provides further evidence about the risk of cross-reactivity between BL classes. These results support the practice of using a BL from an alternative class for patients in need of gram-negative antibiotic coverage. Copyright © 2015 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

Management of post-neurosurgical meningitis: narrative review.

PubMed

Hussein, K; Bitterman, R; Shofty, B; Paul, M; Neuberger, A

2017-09-01

Infections complicating neurosurgery pose unacceptable mortality and morbidity. To summarize what is known about the epidemiology, diagnosis and treatment of post-neurosurgical meningitis (PNM). PubMed, references of identified studies and reviews, and personal experience when evidence was lacking. The incidence and pathogen distribution of PNM is highly variable. A shift towards Gram-negative bacteria has been observed with use of antibiotic prophylaxis and antibiotic-coated devices directed mainly against Gram-positive bacteria. However, knowledge of the local epidemiology is necessary to treat PNM. The diagnosis of PNM is difficult because, unlike community-acquired meningitis, symptoms are less specific; patients are ill at baseline and many neurosurgical conditions mimic meningitis and cause cerebrospinal fluid (CSF) abnormalities. Pivotal CSF findings for diagnosis of PNM are the CSF glucose, CSF lactate and Gram stain. CSF leucocyte counts are not specific in PNM. Current diagnostic capabilities leave a non-negligible category of patients with microbiologically negative, uncertain diagnosis of PNM. There is no high-quality evidence on several cardinal issues in PNM management, including the effectiveness of intraventricular or intrathecal (IV/IT) antibiotics, effectiveness of dual antibiotic therapy for multidrug-resistant Gram-negative bacteria; clinical benefit of routine therapeutic drug monitoring; and safest timing of shunt replacement. Some data point to a potential benefit of IV/IT antibiotic treatment, mainly for PNM caused by carbapenem-resistant Gram-negative bacteria. Carbapenem-colistin combination therapy is suggested for PNM caused by carbapenem-resistant Gram-negative bacteria with a carbapenem MIC ≤8 mg/L. Guiding the optimal management of PNM will necessitate collaborative multicentre efforts and unique study designs. Copyright © 2017 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Prevotella histicola sp. nov., isolated from the human oral cavity.

PubMed

Downes, Julia; Hooper, Samuel J; Wilson, Melanie J; Wade, William G

2008-08-01

Three strains of anaerobic, variably pigmenting, Gram-negative bacilli isolated from human oral mucosal tissue were subjected to a comprehensive range of phenotypic and genotypic tests and were found to comprise a homogeneous group. 16S rRNA gene sequence analysis and DNA-DNA hybridization revealed that the strains constituted a novel group within the genus Prevotella, being most closely related to Prevotella melaninogenica and Prevotella veroralis. A novel species, Prevotella histicola sp. nov., is proposed to accommodate these strains. Prevotella histicola is saccharolytic and produces acetic acid and succinic acid as major end products of fermentation and trace to minor amounts of isovaleric acid and lactic acid. The G+C content of the DNA of the type strain is 43 mol%. The type strain of Prevotella histicola is T05-04T (=DSM 19854T=CCUG 55407T).

Regulatory RNAs in Bacillus subtilis: a Gram-Positive Perspective on Bacterial RNA-Mediated Regulation of Gene Expression.

PubMed

Mars, Ruben A T; Nicolas, Pierre; Denham, Emma L; van Dijl, Jan Maarten

2016-12-01

Bacteria can employ widely diverse RNA molecules to regulate their gene expression. Such molecules include trans-acting small regulatory RNAs, antisense RNAs, and a variety of transcriptional attenuation mechanisms in the 5' untranslated region. Thus far, most regulatory RNA research has focused on Gram-negative bacteria, such as Escherichia coli and Salmonella. Hence, there is uncertainty about whether the resulting insights can be extrapolated directly to other bacteria, such as the Gram-positive soil bacterium Bacillus subtilis. A recent study identified 1,583 putative regulatory RNAs in B. subtilis, whose expression was assessed across 104 conditions. Here, we review the current understanding of RNA-based regulation in B. subtilis, and we categorize the newly identified putative regulatory RNAs on the basis of their conservation in other bacilli and the stability of their predicted secondary structures. Our present evaluation of the publicly available data indicates that RNA-mediated gene regulation in B. subtilis mostly involves elements at the 5' ends of mRNA molecules. These can include 5' secondary structure elements and metabolite-, tRNA-, or protein-binding sites. Importantly, sense-independent segments are identified as the most conserved and structured potential regulatory RNAs in B. subtilis. Altogether, the present survey provides many leads for the identification of new regulatory RNA functions in B. subtilis. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Regulatory RNAs in Bacillus subtilis: a Gram-Positive Perspective on Bacterial RNA-Mediated Regulation of Gene Expression

PubMed Central

Mars, Ruben A. T.; Nicolas, Pierre; Denham, Emma L.

2016-01-01

SUMMARY Bacteria can employ widely diverse RNA molecules to regulate their gene expression. Such molecules include trans-acting small regulatory RNAs, antisense RNAs, and a variety of transcriptional attenuation mechanisms in the 5′ untranslated region. Thus far, most regulatory RNA research has focused on Gram-negative bacteria, such as Escherichia coli and Salmonella. Hence, there is uncertainty about whether the resulting insights can be extrapolated directly to other bacteria, such as the Gram-positive soil bacterium Bacillus subtilis. A recent study identified 1,583 putative regulatory RNAs in B. subtilis, whose expression was assessed across 104 conditions. Here, we review the current understanding of RNA-based regulation in B. subtilis, and we categorize the newly identified putative regulatory RNAs on the basis of their conservation in other bacilli and the stability of their predicted secondary structures. Our present evaluation of the publicly available data indicates that RNA-mediated gene regulation in B. subtilis mostly involves elements at the 5′ ends of mRNA molecules. These can include 5′ secondary structure elements and metabolite-, tRNA-, or protein-binding sites. Importantly, sense-independent segments are identified as the most conserved and structured potential regulatory RNAs in B. subtilis. Altogether, the present survey provides many leads for the identification of new regulatory RNA functions in B. subtilis. PMID:27784798

Prevalence of Bacterial Vaginosis and Associated Risk Factors among Women Complaining of Genital Tract Infection.

PubMed

Bitew, Adane; Abebaw, Yeshiwork; Bekele, Delayehu; Mihret, Amete

2017-01-01

Bacterial vaginosis is a global concern due to the increased risk of acquisition of sexually transmitted infections. To determine the prevalence of bacterial vaginosis and bacteria causing aerobic vaginitis. A cross-sectional study was conducted among 210 patients between September 2015 and July 2016 at St. Paul's Hospital. Gram-stained vaginal swabs were examined microscopically and graded as per Nugent's procedure. Bacteria causing aerobic vaginitis were characterized, and their antimicrobial susceptibility pattern was determined. The overall prevalence of bacterial vaginosis was 48.6%. Bacterial vaginosis was significantly associated with number of pants used per day ( p = 0.001) and frequency of vaginal bathing ( p = 0.045). Of 151 bacterial isolates, 69.5% were Gram-negative and 30.5% were Gram-positive bacteria. The overall drug resistance level of Gram-positive bacteria was high against penicillin, tetracycline, and erythromycin. Cefoxitin and tobramycin were the most active drugs against Gram-positive bacteria. The overall drug resistance level of Gram-negative bacteria was high against tetracycline, ampicillin, and amoxicillin. Amikacin and tobramycin were the most active drugs against Gram-negative bacteria. The prevalence of bacterial vaginosis was high and was affected by individual hygiene. Routine culture of vaginal samples should be performed on patients with vaginitis and the drug susceptibility pattern of each isolate should be determined.

Changes in the Microflora of Haddock Fillets and Shucked Soft-Shelled Clams After Irradiation with Co60 Gamma Rays and Storage at 0 C and 6 C

PubMed Central

Masurovsky, E. B.; Voss, J. S.; Goldblith, S. A.

1963-01-01

Frequency distribution patterns of aerobic-facultative microflora, obtained by random selection of macrocolonies from samples of haddock fillets and shucked soft-shelled clams before and after treatment with doses of from 50,000 to 800,000 rad of Co60 gamma rays, are presented, with analyses and interpretation. The data showed that a decided change occurred in the constitution of the microbial populations of both products: from a mixed gram-negative—gram-positive flora to a predominantly gram-positive flora immediately after irradiation. The great majority of these surviving microorganisms were micrococci, sporeforming bacilli, and certain yeasts, molds, and actinomyces. During storage at refrigeration temperatures above freezing, the microflora changed from the descendants of the more radioresistant gram-positive species to the more prolific gram-negative psychrophilic species that flourish at these low temperatures. Micrococci and gram-positive rods declined somewhat during the rise of the actively proteolytic-lipolytic pseudomonads and related species, but still remained at high enough levels to contribute significantly to the spoilage observed at different times in storage. The eventual spoilage of haddock fillets was characterized by discoloration of the cream-white tissue with water-soluble yellow, green, and red bacterial pigments; degradation of the tissue, by proteolytic and other microbial enzymes, to a watery, flaccid mass; and formation of volatile compounds that smelled putrid, rancid, and generally foul and pungent. Shucked soft-shelled clams displayed a different spoilage pattern, changing to a variegated brown-gray and forming a matted or loose gelatinous mass from which arose stale, acrid, putrid, sulfurous odors. These differences are believed to be attributable to the varied biochemical nature of the tissues involved, the Eh potential within the tissues, the time sequence of microfloral change, and the species and types of microorganisms associated with each product. Staphylococci were present in small numbers in some of the samples tested, but did not appear to be species of public health significance. Gram-negative enteric rods were also encountered, but were considered to be of doubtful public health importance. More detailed investigations, designed to study the effects of Co60 gamma radiations on such microbes in sea foods, would be useful in assessing the problem further. PMID:13933509

Incidence of Carbapenem-Resistant Gram Negatives in Italian Transplant Recipients: A Nationwide Surveillance Study

PubMed Central

Lanini, Simone; Costa, Alessandro Nanni; Puro, Vincenzo; Procaccio, Francesco; Grossi, Paolo Antonio; Vespasiano, Francesca; Ricci, Andrea; Vesconi, Sergio; Ison, Michael G.; Carmeli, Yehuda; Ippolito, Giuseppe

2015-01-01

Background Bacterial infections remain a challenge to solid organ transplantation. Due to the alarming spread of carbapenem-resistant gram negative bacteria, these organisms have been frequently recognized as cause of severe infections in solid organ transplant recipients. Methods and Findings Between 15 May and 30 September 2012 we enrolled 887 solid organ transplant recipients in Italy with the aim to describe the epidemiology of gram negative bacteria spreading, to explore potential risk factors and to assess the effect of early isolation of gram negative bacteria on recipients’ mortality during the first 90 days after transplantation. During the study period 185 clinical isolates of gram negative bacteria were reported, for an incidence of 2.39 per 1000 recipient-days. Positive cultures for gram negative bacteria occurred early after transplantation (median time 26 days; incidence rate 4.33, 1.67 and 1.14 per 1,000 recipient-days in the first, second and third month after SOT, respectively). Forty-nine of these clinical isolates were due to carbapenem-resistant gram negative bacteria (26.5%; incidence 0.63 per 1000 recipient-days). Carbapenems resistance was particularly frequent among Klebsiella spp. isolates (49.1%). Recipients with longer hospital stay and those who received either heart or lung graft were at the highest risk of testing positive for any gram negative bacteria. Moreover recipients with longer hospital stay, lung recipients and those admitted to hospital for more than 48h before transplantation had the highest probability to have culture(s) positive for carbapenem-resistant gram negative bacteria. Forty-four organ recipients died (0.57 per 1000 recipient-days) during the study period. Recipients with at least one positive culture for carbapenem-resistant gram negative bacteria had a 10.23-fold higher mortality rate than those who did not. Conclusion The isolation of gram-negative bacteria is most frequent among recipient with hospital stays >48 hours prior to transplant and in those receiving either heart or lung transplants. Carbapenem-resistant gram negative isolates are associated with significant mortality. PMID:25835018

Diversity of Innate Immune Recognition Mechanism for Bacterial Polymeric meso-Diaminopimelic Acid-type Peptidoglycan in Insects

PubMed Central

Yu, Yang; Park, Ji-Won; Kwon, Hyun-Mi; Hwang, Hyun-Ok; Jang, In-Hwan; Masuda, Akiko; Kurokawa, Kenji; Nakayama, Hiroshi; Lee, Won-Jae; Dohmae, Naoshi; Zhang, Jinghai; Lee, Bok Luel

2010-01-01

In Drosophila, the synthesis of antimicrobial peptides in response to microbial infections is under the control of the Toll and immune deficiency (Imd) signaling pathway. The Toll signaling pathway responds mainly to the lysine-type peptidoglycan of Gram-positive bacteria and fungal β-1,3-glucan, whereas the Imd pathway responds to the meso-diaminopimelic acid (DAP)-type peptidoglycan of Gram-negative bacteria and certain Gram-positive bacilli. Recently we determined the activation mechanism of a Toll signaling pathway biochemically using a large beetle, Tenebrio molitor. However, DAP-type peptidoglycan recognition mechanism and its signaling pathway are still unclear in the fly and beetle. Here, we show that polymeric DAP-type peptidoglycan, but not its monomeric form, formed a complex with Tenebrio peptidoglycan recognition protein-SA, and this complex activated the three-step proteolytic cascade to produce processed Spätzle, a Toll receptor ligand, and induced Drosophila defensin-like antimicrobial peptide in Tenebrio larvae similarly to polymeric lysine-type peptidoglycan. Monomeric DAP-type peptidoglycan induced Drosophila diptericin-like antimicrobial peptide in Tenebrio hemocytes. In addition, both polymeric and monomeric DAP-type peptidoglycans induced expression of Tenebrio peptidoglycan recognition protein-SC2, which is DAP-type peptidoglycan-selective N-acetylmuramyl-l-alanine amidase that functions as a DAP-type peptidoglycan scavenger, appearing to function as a negative regulator of the DAP-type peptidoglycan signaling by cleaving DAP-type peptidoglycan in Tenebrio larvae. Taken together, these results demonstrate that molecular recognition mechanism for polymeric DAP-type peptidoglycan is different between Tenebrio larvae and Drosophila adults, providing biochemical evidences of biological diversity of innate immune responses in insects. PMID:20702416

DIFFERENCES IN CULTURAL YIELD OF MYCOBACTERIUM TUBERCULOSIS ON MEDIA PREPARED USING COMMERCIAL AND HOUSEHOLD EGGS.

PubMed

Noori, Muhammad Yahya; AJi, Zaheer; Khan, Ghazala; Sharafat, Shaheen; Masroor, Muhammad

2015-01-01

Mycobacterial culture is considered as the gold standard for TB diagnosis. It is performed on egg-based media using commercially available eggs to grow Mycobacteria from clinical samples. These eggs are known to contain high concentration of antibiotics, including fluoroquinolones, given to chicken to prevent early mortality. This study was performed to compare Mycobacterial growth on media prepared from commercial and antibiotic free household eggs. Sputum samples from negative (No bacilli in 100 oil immersion field), scanty (1-9 AFB in 100 fields), 1+ (10-99 bacilli per field), 2+ (1-10 bacilli per field) and 3+ (>10 bacilli per field) were inoculated dually on Ogawa medium prepared from commercial and household eggs. Tubes were inspected every fourth day for the appearance of colonies till 60 days. Data tabulations and statistical analysis (F test for variation and unpaired Student's t test) were performed on Microsoft Excel. One microscopically negative sample showed growth on media prepared from household eggs, while all were negative on that prepared from commercial eggs. There were significant differences in time to culture positivity for samples graded 1+ (p = 0.02), 2+ (p = 0.002) and 3+ (p = 0.0003). Commercial eggs containing antibiotics can be a source of false negativity in cultures especially in microscopically negative samples. This can be of special concern in HIV patients who have high smear negativity. It is therefore important to either develop provision of antibiotic free eggs for media preparation or to develop and validate other laboratory investigations for smear negative TB patients.

Comparative genomic analyses of Clavibacter michiganensis subsp. insidiosus and aggressiveness on Medicago truncatula

USDA-ARS?s Scientific Manuscript database

Clavibacter michiganensis is the most economically important gram-positive bacterial plant pathogen with subspecies that cause serious diseases of maize, wheat, tomato, potato, and alfalfa. Much less is known about pathogenesis involving gram-positive plant pathogens than is known for gram-negative ...

Drug-resistant gram-negative uropathogens: A review.

PubMed

Khoshnood, Saeed; Heidary, Mohsen; Mirnejad, Reza; Bahramian, Aghil; Sedighi, Mansour; Mirzaei, Habibollah

2017-10-01

Urinary tract infection(UTI) caused by Gram-negative bacteria is the second most common infectious presentation in community medical practice. Approximately 150 million people are diagnosed with UTI each year worldwide. Drug resistance in Gram-negative uropathogens is a major global concern which can lead to poor clinical outcomes including treatment failure, development of bacteremia, requirement for intravenous therapy, hospitalization, and extended length of hospital stay. The mechanisms of drug resistance in these bacteria are important due to they are often not identified by routine susceptibility tests and have an exceptional potential for outbreaks. Treatment of UTIs depends on the access to effective drugs, which is now threatened by antibiotic resistant Gram-negative uropathogens. Although several effective antibiotics with activity against highly resistant Gram-negatives are available, there is not a unique antibiotic with activity against the high variety of resistance. Therefore, antimicrobial susceptibility tests, correlation between clinicians and laboratories, development of more rapid diagnostic methods, and continuous monitoring of drug resistance are urgent priorities. In this review, we will discuss about the current global status of drug-resistant Gram-negative uropathogens and their mechanisms of drug resistance to provide new insights into their treatment options. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Evaluation of the MicroScan ESBL plus confirmation panel for detection of extended-spectrum beta-lactamases in clinical isolates of oxyimino-cephalosporin-resistant Gram-negative bacteria.

PubMed

Stürenburg, Enno; Lang, Melanie; Horstkotte, Matthias A; Laufs, Rainer; Mack, Dietrich

2004-11-01

We aimed to assess the performance of the MicroScan ESBL plus confirmation panel using a series of 87 oxyimino-cephalosporin-resistant Gram-negative bacilli of various species. Organisms tested included 57 extended-spectrum beta-lactamase (ESBL) strains comprising Enterobacter aerogenes (3), Enterobacter cloacae (10), Escherichia coli (11), Klebsiella pneumoniae (26), Klebsiella oxytoca (3) and Proteus mirabilis (4). Also included were 30 strains resistant to oxyimino cephalosporins but lacking ESBLs, which were characterized with other resistance mechanisms, such as inherent clavulanate susceptibility in Acinetobacter spp. (4), hyperproduction of AmpC enzyme in Citrobacter freundii (2), E. aerogenes (3), E. cloacae (3), E. coli (4), Hafnia alvei (1) and Morganella morganii (1), production of plasmid-mediated AmpC beta-lactamase in K. pneumoniae (3) and E. coli (3) or hyperproduction of K1 enzyme in K. oxytoca (6). The MicroScan MIC-based clavulanate synergy correctly classified 50 of 57 ESBL strains as ESBL-positive and 23 of 30 non-ESBL strains as ESBL-negative (yielding a sensitivity of 88% and a specificity of 76.7%, respectively). False negatives among ESBL producers were highest with Enterobacter spp. due to masking interactions between ESBL and AmpC beta-lactamases. False-positive classifications occurred in two Acinetobacter spp., one E. coli producing plasmid-mediated AmpC beta-lactamase and two K. oxytoca hyperproducing their chromosomal K1 beta-lactamase. The MicroScan clavulanate synergy test proved to be a valuable tool for ESBL confirmation. However, this test has limitations in detecting ESBLs in Enterobacter spp. and in discriminating ESBL-related resistance from the K1 enzyme and from inherent clavulanate susceptibility in Acinetobacter spp.

Red blood cell distribution width is an independent predictor of mortality in patients with gram-negative bacteremia.

PubMed

Ku, Nam Su; Kim, Hye-Won; Oh, Hyung Jung; Kim, Yong Chan; Kim, Min Hyung; Song, Je Eun; Oh, Dong Hyun; Ahn, Jin Young; Kim, Sun Bean; Jeong, Su Jin; Han, Sang Hoon; Kim, Chang Oh; Song, Young Goo; Kim, June Myung; Choi, Jun Yong

2012-08-01

Red blood cell distribution width (RDW) is known to be a predictor of severe morbidity and mortality in some chronic diseases such as congestive heart failure. However, to our knowledge, little is known about RDW as a predictor of mortality in patients with Gram-negative bacteremia, a major nosocomial cause of intra-abdominal infections, urinary tract infections, and primary bacteremia. Therefore, we investigated whether RDW is an independent predictor of mortality in patients with Gram-negative bacteremia. Clinical characteristics, laboratory parameters, and outcomes of 161 patients with Gram-negative bacteremia from November 2010 to March 2011 diagnosed at Severance Hospital, Yonsei University College of Medicine, Seoul, Korea, were retrospectively analyzed. The main outcome measure was 28-day all-cause mortality. The 28-day mortality rate was significantly higher in the increased RDW group compared with the normal RDW group (P < 0.001). According to multivariate Cox proportional hazard analysis, RDW levels at the onset of bacteremia (per 1% increase, P = 0.036), the Charlson index (per 1-point increase, P < 0.001), and the Sequential Organ Failure Assessment score (per 1-point increase, P = 0.001) were independent risk factors for 28-day mortality. Moreover, the nonsurvivor group had significantly higher RDW levels 72 h after the onset of bacteremia than did the survivor group (P = 0.001). In addition, the area under the curve of RDW at the onset of bacteremia, the 72-h RDW, and the Sequential Organ Failure Assessment score for 28-day mortality were 0.764 (P = 0.001), 0.802 (P < 0.001), and 0.703 (P = 0.008), respectively. Red blood cell distribution width at the onset of bacteremia was an independent predictor of mortality in patients with Gram-negative bacteremia. Also, 72-h RDW could be a predictor for all-cause mortality in patients with Gram-negative bacteremia.

The incidence and clinical implication of sputum with positive acid-fast bacilli smear but negative in mycobacterial culture in a tertiary referral hospital in South Korea.

PubMed

Lee, Jae Seok; Kim, Eui-Chong; Joo, Sei Ick; Lee, Sang-Min; Yoo, Chul-Gyu; Kim, Young Whan; Han, Sung Koo; Shim, Young-Soo; Yim, Jae-Joon

2008-10-01

Although it is not rare to find sputum that is positive acid-fast bacilli (AFB) smear but subsequent culture fails to isolate mycobacteria in clinical practice, the incidence and clinical implication of those sputa from new patients has not been clearly elucidated. The aim of this study was to determine the incidence and clinical implication of sputum with positive AFB smear but negative in mycobacterial culture. All sputa that were positive AFB smear requested during diagnostic work up for new patients visiting Seoul National University Hospital from 1 January 2005 through 31 December 2006 were included. Sputa producing a positive AFB smear but negative mycobacterial culture were classified into one of four categories: laboratory failure to isolate mycobacteria, false positive AFB smear, pathogen may show a positive AFB smear other than mycobacteria, and indeterminate results. Out of 447 sputa with a positive AFB smear, 29 (6.5%) failed to culture any organism. Among these 29 sputa, 18 were caused by laboratory failure to isolate mycobacteria, six were false positive smears, and five indeterminate. Although most sputum with a positive AFB smear but negative culture could be classified as a laboratory failure, clinicians should consider the possibility of false positive AFB smear.

Gram-negative prosthetic joint infection: outcome of a debridement, antibiotics and implant retention approach. A large multicentre study.

PubMed

Rodríguez-Pardo, D; Pigrau, C; Lora-Tamayo, J; Soriano, A; del Toro, M D; Cobo, J; Palomino, J; Euba, G; Riera, M; Sánchez-Somolinos, M; Benito, N; Fernández-Sampedro, M; Sorli, L; Guio, L; Iribarren, J A; Baraia-Etxaburu, J M; Ramos, A; Bahamonde, A; Flores-Sánchez, X; Corona, P S; Ariza, J

2014-11-01

We aim to evaluate the epidemiology and outcome of gram-negative prosthetic joint infection (GN-PJI) treated with debridement, antibiotics and implant retention (DAIR), identify factors predictive of failure, and determine the impact of ciprofloxacin use on prognosis. We performed a retrospective, multicentre, observational study of GN-PJI diagnosed from 2003 through to 2010 in 16 Spanish hospitals. We define failure as persistence or reappearance of the inflammatory joint signs during follow-up, leading to unplanned surgery or repeat debridement>30 days from the index surgery related death, or suppressive antimicrobial therapy. Parameters predicting failure were analysed with a Cox regression model. A total of 242 patients (33% men; median age 76 years, interquartile range (IQR) 68-81) with 242 episodes of GN-PJI were studied. The implants included 150 (62%) hip, 85 (35%) knee, five (2%) shoulder and two (1%) elbow prostheses. There were 189 (78%) acute infections. Causative microorganisms were Enterobacteriaceae in 78%, Pseudomonas spp. in 20%, and other gram-negative bacilli in 2%. Overall, 19% of isolates were ciprofloxacin resistant. DAIR was used in 174 (72%) cases, with an overall success rate of 68%, which increased to 79% after a median of 25 months' follow-up in ciprofloxacin-susceptible GN-PJIs treated with ciprofloxacin. Ciprofloxacin treatment exhibited an independent protective effect (adjusted hazard ratio (aHR) 0.23; 95% CI, 0.13-0.40; p<0.001), whereas chronic renal impairment predicted failure (aHR, 2.56; 95% CI, 1.14-5.77; p 0.0232). Our results confirm a 79% success rate in ciprofloxacin-susceptible GN-PJI treated with debridement, ciprofloxacin and implant retention. New therapeutic strategies are needed for ciprofloxacin-resistant PJI. © 2014 The Authors Clinical Microbiology and Infection © 2014 European Society of Clinical Microbiology and Infectious Diseases.

Performance of two resin-containing blood culture media in detection of bloodstream infections and in direct matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) broth assays for isolate identification: clinical comparison of the BacT/Alert Plus and Bactec Plus systems.

PubMed

Fiori, Barbara; D'Inzeo, Tiziana; Di Florio, Viviana; De Maio, Flavio; De Angelis, Giulia; Giaquinto, Alessia; Campana, Lara; Tanzarella, Eloisa; Tumbarello, Mario; Antonelli, Massimo; Sanguinetti, Maurizio; Spanu, Teresa

2014-10-01

We compared the clinical performances of the BacT/Alert Plus (bioMérieux) and Bactec Plus (Becton Dickinson) aerobic and anaerobic blood culture (BC) media with adsorbent polymeric beads. Patients ≥ 16 years old with suspected bloodstream infections (BSIs) were enrolled in intensive care units and infectious disease wards. A single 40-ml blood sample was collected from each and used to inoculate (10 ml/bottle) one set of BacT/Alert Plus cultures and one set of Bactec Plus cultures, each set consisting of one aerobic and one anaerobic bottle. Cultures were incubated ≤ 5 days in the BacT/Alert 3D and Bactec FX instruments, respectively. A total of 128 unique BSI episodes were identified based on the recovery of clinically significant growth in 212 aerobic cultures (106 BacT/Alert and 106 Bactec) and 151 anaerobic cultures (82 BacT/Alert and 69 Bactec). The BacT/Alert aerobic medium had higher recovery rates for Gram-positive cocci (P = 0.024), whereas the Bactec aerobic medium was superior for recovery of Gram-negative bacilli (P = 0.006). BacT/Alert anaerobic medium recovery rates exceeded those of the Bactec anaerobic medium for total organisms (P = 0.003), Gram-positive cocci (P = 0.013), and Escherichia coli (P = 0.030). In terms of capacity for diagnosing the 128 septic episodes, the BacT/Alert and Bactec sets were comparable, although the former sets diagnosed more BSIs caused by Gram-positive cocci (P = 0.008). They also allowed earlier identification of coagulase-negative staphylococcal growth (mean, 2.8 h; P = 0.003) and growth in samples from patients not on antimicrobial therapy that yielded positive results (mean, 1.3 h; P < 0.001). Similarly high percentages of microorganisms in BacT/Alert and Bactec cultures (93.8% and 93.3%, respectively) were identified by direct matrix-assisted laser desorption ionization-time of flight mass spectrometry assay of BC broths. The BacT/Alert Plus media line appears to be a reliable, timesaving tool for routine detection of BSIs in the population we studied, although further studies are needed to evaluate their performance in other settings. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Pyramidobacter piscolens gen. nov., sp. nov., a member of the phylum 'Synergistetes' isolated from the human oral cavity.

PubMed

Downes, Julia; Vartoukian, Sonia R; Dewhirst, Floyd E; Izard, Jacques; Chen, Tsute; Yu, Wen-Han; Sutcliffe, Iain C; Wade, William G

2009-05-01

Four strains of anaerobic, Gram-negative bacilli isolated from the human oral cavity were subjected to a comprehensive range of phenotypic and genotypic tests and were found to comprise a homogeneous group distinct from any species with validly published names. 16S rRNA and 23S rRNA gene sequence analyses and DNA-DNA reassociation data revealed that the strains constituted a novel group within the phylum 'Synergistetes' and were most closely related to Jonquetella anthropi. Two libraries of randomly cloned DNA were prepared from strain W5455(T) and were sequenced to provide a genome survey as a resource for metagenomic studies. A new genus and novel species, Pyramidobacter piscolens gen. nov., sp. nov., is proposed to accommodate these strains. The genus Pyramidobacter comprises strains that are anaerobic, non-motile, asaccharolytic bacilli that produce acetic and isovaleric acids and minor to trace amounts of propionic, isobutyric, succinic and phenylacetic acids as end products of metabolism. P. piscolens gen. nov., sp. nov. produced hydrogen sulphide but was otherwise largely biochemically unreactive. Growth was stimulated by the addition of glycine to broth media. The G+C content of the DNA of the type strain was 59 mol%. The type strain of Pyramidobacter piscolens sp. nov. is W5455(T) (=DSM 21147(T)=CCUG 55836(T)).

Pyramidobacter piscolens gen. nov., sp. nov., a member of the phylum ‘Synergistetes’ isolated from the human oral cavity

PubMed Central

Downes, Julia; Vartoukian, Sonia R.; Dewhirst, Floyd E.; Izard, Jacques; Chen, Tsute; Yu, Wen-Han; Sutcliffe, Iain C.; Wade, William G.

2009-01-01

Four strains of anaerobic, Gram-negative bacilli isolated from the human oral cavity were subjected to a comprehensive range of phenotypic and genotypic tests and were found to comprise a homogeneous group distinct from any species with validly published names. 16S rRNA and 23S rRNA gene sequence analyses and DNA–DNA reassociation data revealed that the strains constituted a novel group within the phylum ‘Synergistetes’ and were most closely related to Jonquetella anthropi. Two libraries of randomly cloned DNA were prepared from strain W5455T and were sequenced to provide a genome survey as a resource for metagenomic studies. A new genus and novel species, Pyramidobacter piscolens gen. nov., sp. nov., is proposed to accommodate these strains. The genus Pyramidobacter comprises strains that are anaerobic, non-motile, asaccharolytic bacilli that produce acetic and isovaleric acids and minor to trace amounts of propionic, isobutyric, succinic and phenylacetic acids as end products of metabolism. P. piscolens gen. nov., sp. nov. produced hydrogen sulphide but was otherwise largely biochemically unreactive. Growth was stimulated by the addition of glycine to broth media. The G+C content of the DNA of the type strain was 59 mol%. The type strain of Pyramidobacter piscolens sp. nov. is W5455T (=DSM 21147T=CCUG 55836T). PMID:19406777

Outbreak of meningitis in weaner pigs caused by unidentified asaccharolytic gram-negative bacterium.

PubMed Central

Mohan, K; Holmes, B; Kock, N; Muvavarirwa, P

1996-01-01

Several organisms are known to cause outbreaks of meningitis in pigs, with Haemophilus species being the most frequently implicated. We report such an outbreak in which necropsied pigs manifested an unusual combination of meningitis, tracheitis, and bronchitis. The causative agent appeared to be an asaccharolytic gram-negative nonfermentative bacterium whose classification has yet to be determined. The organism was isolated from the brain and was extremely capnophilic, growing in air only after several serial subcultures. PMID:8815112

TLR4-mediated podosome loss discriminates gram-negative from gram-positive bacteria in their capacity to induce dendritic cell migration and maturation.

PubMed

van Helden, Suzanne F G; van den Dries, Koen; Oud, Machteld M; Raymakers, Reinier A P; Netea, Mihai G; van Leeuwen, Frank N; Figdor, Carl G

2010-02-01

Chronic infections are caused by microorganisms that display effective immune evasion mechanisms. Dendritic cell (DC)-dependent T cell-mediated adaptive immunity is one of the mechanisms that have evolved to prevent the occurrence of chronic bacterial infections. In turn, bacterial pathogens have developed strategies to evade immune recognition. In this study, we show that gram-negative and gram-positive bacteria differ in their ability to activate DCs and that gram-negative bacteria are far more effective inducers of DC maturation. Moreover, we observed that only gram-negative bacteria can induce loss of adhesive podosome structures in DCs, a response necessary for the induction of effective DC migration. We demonstrate that the ability of gram-negative bacteria to trigger podosome turnover and induce DC migration reflects their capacity to selectively activate TLR4. Examining mice defective in TLR4 signaling, we show that this DC maturation and migration are mainly Toll/IL-1 receptor domain-containing adaptor-inducing IFNbeta-dependent. Furthermore, we show that these processes depend on the production of PGs by these DCs, suggesting a direct link between TLR4-mediated signaling and arachidonic metabolism. These findings demonstrate that gram-positive and gram-negative bacteria profoundly differ in their capacity to activate DCs. We propose that this inability of gram-positive bacteria to induce DC maturation and migration is part of the armamentarium necessary for avoiding the induction of an effective cellular immune response and may explain the frequent involvement of these pathogens in chronic infections.

Wound infection after median sternotomy during the war in Croatia.

PubMed

Jelić, I; Anić, D; Alfirević, I; Kalinić, S; Ugljen, R; Letica, D; Ante Korda, Z; Vucemilo, I; Bulat, C; Predrijevac, M; Corić, V; Husar, J; Jelic, M; Hulina, D; Depina, I; Dadić, D

1996-12-01

From 1990 to 1994 at Clinical Hospital Center, Zagreb, 1904 median sternotomies were performed for cardiac operations. Patients shared the same intensive care unit (ICU) with the wounded persons, admitted to the hospital from battlefield. Infection developed in 124 patients, an incidence of 6.51%. Methicillin resistant Staphylococcus aureus (MRSA) was isolated from 90, methicillin resistant Staphylococcus epidermidis (MRSE) from 19, and gram negative bacilli (GNB) from 56 patients, Pseudomonas aeruginosa in 2, and Clostridium pneumoniae in 1 case. Ninety-six patients (5.04%) developed superficial localized infection of subcutaneous tissues and they were treated with frequent dressing changes with antibiotic-soaked gauze in combination with systemic antibiotics. Twenty-eight patients (1.47%) developed mediastinitis and sternal dehiscence. They were treated by operative debridement followed by reclosure of the sternum with continuous antibiotic irrigation. We obtained satisfactory results with our method of closure of sternum which is a modification of Robicsek's technique. Nine of them required further operation. In seven cases we performed muscle flaps and in two omentoplasty. One hundred and twenty patients were discharged in satisfactory condition. The uncontrolled mediastinal sepsis caused death in 4 patients. Higher infection rate after median sternotomy during 1991 and 1992 could be possibly explained with the war circumstances in Croatia, and especially with MRSA strain becoming endemic in surgical ICU.

[Septic shock Fusobacterium necrophorum from origin gynecological at complicated an acute respiratory distress syndrome: a variant of Lemierre's syndrome].

PubMed

Huynh-Moynot, Sophie; Commandeur, Diane; Danguy des Déserts, Marc; Drouillard, Isabelle; Leguen, Patrick; Ould-Ahmed, Mehdi

2011-01-01

We report a case of a female patient of 47 years old who presents in a state of septic shock with acute insufficient respiratory complicated with syndrome of acute respiratory distress, together with a list of abdominal pain and polyarthralgia too. In her case of medical history, it is retained that she has had a intra-uterine device since 6 years without medical follow up. The initial thoraco-abdomino-pelvic scan shows a left ovarian vein thrombosis, as well as the opaqueness alveolus diffused interstitiel bilaterally and an aspect of ileitis. The IUD is taken off because of sudden occuring of purulent leucorrhoea. This results in a clinical and paraclinical improvement, whereas aminopenicillin was administered to the patient since 1 week. The microbiological blood test allows to put in evidence Fusobacterium necrophorum found in a blood culture and is sensitive to the amoxicilline-acide clavulanique and metronidazole. Isolation of this bacteria, classically found in Lemierre's syndrome, allowed to explain the multilfocalization of the symtoms and the list of pain. The whole concerns about a variant of Lemierre's syndrom: a state of septic shock secondary then caused by the anaerobic Gram negative bacilli, which is a commensal bacteria of the female genital tractus, complicated of septic emboli typical.

Changing epidemiology of central venous catheter-related bloodstream infections: increasing prevalence of Gram-negative pathogens.

PubMed

Marcos, Miguel; Soriano, Alex; Iñurrieta, Amaia; Martínez, José A; Romero, Alberto; Cobos, Nazaret; Hernández, Cristina; Almela, Manel; Marco, Francesc; Mensa, Josep

2011-09-01

Gram-positive microorganisms have been the predominant pathogens in central venous catheter-related bloodstream infections (CRBSIs). Recent guidelines recommend empirical therapy according to this and restrict coverage for Gram-negatives to specific circumstances. This study aimed to analyse the epidemiological changes in CRBSIs over the 1991-2008 period and to analyse predictors of Gram-negative CRBSIs. A prospectively collected cohort of patients with confirmed CRBSIs was analysed. Strains isolated and antimicrobial susceptibility, as well as clinical and demographic variables were recorded. Differences observed during the study period were analysed by means of a χ² trend test and factors associated with Gram-negative CRBSIs by means of multivariable analysis. Between 1991 and 2008, 1129 episodes of monomicrobial CRBSIs were recorded. There was an increase in the incidence of CRBSIs, from 0.10 (1991-92) to 0.31 (2007-08) episodes/1000 patient-days. A significant increase in the number of Gram-negative strains among the total isolates was also found, from 3 (4.7%) in 1991-92 to 70 (40.23%) in 2007-08, with a parallel decrease in the percentage of Gram-positives. Solid organ transplantation, prior use of penicillins and hospital stay longer than 11 days were independently associated with a significantly higher risk of Gram-negative CRBSIs, while cirrhosis, diabetes and use of quinolones were associated with a higher risk of Gram-positives. Gram-negative strains are an increasing cause of CRBSIs, reaching a prevalence of 40% in the 2007-08 period in our hospital. If this trend is confirmed in other centres, a broad-spectrum empirical therapy should be considered in managing these infections.

Prevalence of Bacterial Vaginosis and Associated Risk Factors among Women Complaining of Genital Tract Infection

PubMed Central

Abebaw, Yeshiwork; Bekele, Delayehu; Mihret, Amete

2017-01-01

Background Bacterial vaginosis is a global concern due to the increased risk of acquisition of sexually transmitted infections. Objectives To determine the prevalence of bacterial vaginosis and bacteria causing aerobic vaginitis. Methods A cross-sectional study was conducted among 210 patients between September 2015 and July 2016 at St. Paul's Hospital. Gram-stained vaginal swabs were examined microscopically and graded as per Nugent's procedure. Bacteria causing aerobic vaginitis were characterized, and their antimicrobial susceptibility pattern was determined. Results The overall prevalence of bacterial vaginosis was 48.6%. Bacterial vaginosis was significantly associated with number of pants used per day (p = 0.001) and frequency of vaginal bathing (p = 0.045). Of 151 bacterial isolates, 69.5% were Gram-negative and 30.5% were Gram-positive bacteria. The overall drug resistance level of Gram-positive bacteria was high against penicillin, tetracycline, and erythromycin. Cefoxitin and tobramycin were the most active drugs against Gram-positive bacteria. The overall drug resistance level of Gram-negative bacteria was high against tetracycline, ampicillin, and amoxicillin. Amikacin and tobramycin were the most active drugs against Gram-negative bacteria. Conclusions The prevalence of bacterial vaginosis was high and was affected by individual hygiene. Routine culture of vaginal samples should be performed on patients with vaginitis and the drug susceptibility pattern of each isolate should be determined. PMID:28831285

The comparison of pyrosequencing molecular Gram stain, culture, and conventional Gram stain for diagnosing orthopaedic infections.

PubMed

Kobayashi, Naomi; Bauer, Thomas W; Tuohy, Marion J; Lieberman, Isador H; Krebs, Viktor; Togawa, Daisuke; Fujishiro, Takaaki; Procop, Gary W

2006-08-01

We have developed a combined real-time PCR and pyrosequencing assay that successfully differentiated the vast majority of gram-positive and gram-negative bacteria when bacterial isolates were tested. The purpose of this study was to evaluate this assay on clinical specimens obtained from orthopedic surgeries, and to prospectively compare the results of "molecular Gram stain" with culture and conventional direct Gram stain. Forty-five surgical specimens were obtained from patients who underwent orthopedic surgery procedures. The DNA was extracted and a set of broad-range PCR primers that targeted a part of the 16S rDNA gene was used for pan-bacterial PCR. The amplicons were submitted for pyrosequencing and the resulting molecular Gram stain characteristics were recorded. Culture and direct Gram staining were performed using standard methods for all cases. Surgical specimens were reviewed histologically for all cases that had a discrepancy between culture and molecular results. There was an 86.7% (39/45) agreement between the traditional and molecular methods. In 12/14 (85.7%) culture-proven cases of bacterial infection, molecular Gram stain characteristics were in agreement with the culture results, while the conventional Gram stain result was in agreement only for five cases (35.7%). In the 31 culture negative cases, 27 cases were also PCR negative, whereas 4 were PCR positive. Three of these were characterized as gram negative and one as gram positive by this molecular method. Molecular determination of the Gram stain characteristics of bacteria that cause orthopedic infections may be achieved, in most instances, by this method. Further studies are necessary to understand the clinical importance of PCR-positive/culture-negative results.

Intestinal perforation in typhoid fever: a historical and state-of-the-art review.

PubMed

Bitar, R; Tarpley, J

1985-01-01

The appropriate therapy for intestinal perforation in typhoid fever has been controversial since the late 1880s. Around the turn of the century, surgery became the established mode of therapy, with a mortality of 69% based on 166 patients in the English-language medical literature, and continued to be the preferred treatment until the advent of chloramphenicol in 1948. At this time the surgical mortality was approximately 50%. Following the recovery of a few patients with perforation treated only with antimicrobial agents (six initially, then eventually 22), nonsurgical therapy became the accepted mode of treatment. This change was never justified and this review demonstrates this. Appropriate therapy is virtually always surgical, usually consisting of simple closure and irrigation. Chloramphenicol alone is inadequate antimicrobial therapy in a patient with perforation and must be supplemented by other antimicrobials directed against enteric aerobic gram-negative bacilli and enteric anaerobes.

A two-hour antibiotic susceptibility test by ATP-bioluminescence.

PubMed

March Rosselló, Gabriel Alberto; García-Loygorri Jordán de Urries, María Cristina; Gutiérrez Rodríguez, María Purificación; Simarro Grande, María; Orduña Domingo, Antonio; Bratos Pérez, Miguel Ángel

2016-01-01

The antibiotic susceptibility test (AST) in Clinical Microbiology laboratories is still time-consuming, and most procedures take 24h to yield results. In this study, a rapid antimicrobial susceptibility test using ATP-bioluminescence has been developed. The design of method was performed using five ATCC collection strains of known susceptibility. This procedure was then validated against standard commercial methods on 10 strains of enterococci, 10 staphylococci, 10 non-fermenting gram negative bacilli, and 13 Enterobacteriaceae from patients. The agreement obtained in the sensitivity between the ATP-bioluminescence method and commercial methods (E-test, MicroScan and VITEK2) was 100%. In summary, the preliminary results obtained in this work show that the ATP-bioluminescence method could provide a fast and reliable AST in two hours. Copyright © 2015 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

Fusobacterial liver abscess: a case report and review of the literature.

PubMed

Jayasimhan, Dilip; Wu, Linus; Huggan, Paul

2017-06-20

Fusobacteriae are facultative anaerobic gram-negative bacilli which cause a range of invasive infections, amongst which pyogenic liver abscesses are rare. We describe a case of Fusobacterium nucleatum liver abscess and review the relevant literature. A 51-year-old lady presented with a 4-day history of abdominal pain, diarrhoea, fever, rigors, and lethargy. Imaging revealed an abscess which was drained. Cultures of the blood and abscess aspirate grew Fusobacterium nucleatum and Prevotella pleuritidis respectively. She achieved full recovery following treatment. A MEDLINE search was undertaken using free-text and Medical Subject Headings (MeSH), keywords "Fusobacterium" and "Liver abscess". Non-English language reports and cases without confirmed growth of Fusobacterium species were excluded. Additional cases were identified by surveying the references of each report and by using the same keywords in a web-based search. Forty-eight cases were identified, 41 in men. The median age was 42.5, with an interquartile range of 33. F. nucleatum and F. necrophorum were in involved in 22 cases each, and 4 cases were not further speciated. Among cases of F. nucleatum liver abscess, nine were attributed to periodontal disease, four to lower gastrointestinal tract disease, one to Lemierre's Syndrome, and eight were considered cryptogenic. All patients treated made a full recovery. Antimicrobial treatment duration ranged from 2 weeks to 6 months with a median of 6 weeks. Fusobacterium nucleatum is an uncommon cause of liver abscess generally associated with good clinical outcomes with contemporary medical and surgical care.

Extended spectrum β-lactamases, carbapenemases and mobile genetic elements responsible for antibiotics resistance in Gram-negative bacteria.

PubMed

El Salabi, Allaaeddin; Walsh, Timothey R; Chouchani, Chedly

2013-05-01

Infectious diseases due to Gram-negative bacteria are a leading cause of morbidity and mortality worldwide. Antimicrobial agents represent one major therapeutic tools implicated to treat these infections. The misuse of antimicrobial agents has resulted in the emergence of resistant strains of Gram-negatives in particular Enterobacteriaceae and non-fermenters; they have an effect not only on a human but on the public health when bacteria use the resistance mechanisms to spread in the hospital environment and to the community outside the hospitals by means of mobile genetic elements. Gram-negative bacteria have become increasingly resistant to antimicrobial agents. They have developed several mechanisms by which they can withstand to antimicrobials, these mechanisms include the production of Extended-spectrum β-lactamases (ESBLs) and carbapenemases, furthermore, Gram-negative bacteria are now capable of spreading such resistance between members of the family Enterobacteriaceae and non-fermenters using mobile genetic elements as vehicles for such resistance mechanisms rendering antibiotics useless. Therefore, addressing the issue of mechanisms of antimicrobial resistance is considered one of most urgent priorities. This review will help to illustrate different resistance mechanisms; ESBLs, carbapenemases encoded by genes carried by mobile genetic elements, which are used by Gram-negative bacteria to escape antimicrobial effect.

Characterization of indoor bioaerosols from a hospital ward in a tropical setting.

PubMed

Sudharsanam, S; Swaminathan, S; Ramalingam, A; Thangavel, G; Annamalai, R; Steinberg, R; Balakrishnan, K; Srikanth, P

2012-06-01

Study was conducted to assess whether temporal variation exists in airborne microbial concentrations of a hospital ward (west-Chennai, India) using active and passive methods, and characterise the microorganisms. Air samples (duplicates) were collected simultaneously using exposed-plate, impingement (BioSampler) and filtration (personal sampling filter cassette loaded with gelatin filter) methods over different periods of the year. Bacterial plates were incubated at 37°C and observed for growth after 48h; fungal plates were incubated at 25°C and 37°C and observed upto 7 days. Microorganisms were identified using standard microbiological procedures. Microbial loads were found to vary with the sampling method. Concentrations of bacteria were higher (exposed-plate: 45-150 CFU/plate; impingement: 1.12E+03-1.6856E+05 CFU/m(3); filtration: 3.788E+03-1.91111E+05 CFU/m(3)) than fungi (exposed-plate: 0-13 CFU/plate; impingement: 0-3.547E+03 CFU/m(3); filtration: 0-1.515E+04 CFU/ m(3)). Coagulase-negative Staphylococci and Micrococci were the predominant Gram-positive cocci in active and passive samples. Enterobacter and Pseudomonas were the predominant Gram-negative bacilli. Among fungi, Aspergillus niger was isolated throughout the year. There was no significant temporal variation in airborne microbial loads irrespective of methods. Exposed-plate method was found to capture microorganisms efficiently with little variation in duplicate samples, suggesting its use in hospitals for preliminary assessment of indoor air quality and determine pathogenic microorganisms due to particle fall-out.

Ventilator-associated pneumonia at a tertiary-care center in a developing country: incidence, microbiology, and susceptibility patterns of isolated microorganisms.

PubMed

Kanafani, Zeina A; Kara, Layla; Hayek, Shady; Kanj, Souha S

2003-11-01

Ventilator-associated pneumonia (VAP) complicates the course of up to 24% of intubated patients. Data from the Middle East are scarce. The objective of this study was to evaluate the incidence, microbiology, and antimicrobial susceptibility patterns of isolated microorganisms in VAP in a developing country. Prospective observational cohort study. The American University of Beirut Medical Center, a tertiary-care center that serves as a major referral center for Lebanon and neighboring countries. All patients admitted to the intensive care and respiratory care units from March to September 2001, and who had been receiving mechanical ventilation for at least 48 hours, were included in the study. Results of samples submitted for culture were recorded and antimicrobial susceptibility testing of isolated pathogens was performed. Seventy patients were entered into the study. The incidence of VAP was 47%. Gram-negative bacilli accounted for 83% of all isolates. The most commonly identified organism was Acinetobacter anitratus, followed by Pseudomonas aeruginosa. Fifty percent of all gram-negative bacterial isolates were classified as antibiotic resistant. Compared with patients without VAP, patients with VAP remained intubated for a longer period and stayed in the intensive care unit longer. VAP was not associated with an increased mortality rate. Compared with other studies, the results from this referral center in Lebanon indicate a higher incidence of VAP and a high prevalence of resistant organisms. These data are relevant because they direct the choice of empiric antibiotic therapy for VAP.

Triton Hodge Test: Improved Protocol for Modified Hodge Test for Enhanced Detection of NDM and Other Carbapenemase Producers

PubMed Central

Pasteran, Fernando; Gonzalez, Lisandro J.; Albornoz, Ezequiel; Bahr, Guillermo; Vila, Alejandro J.

2015-01-01

Accurate detection of carbapenemase-producing Gram-negative bacilli is of utmost importance for the control of nosocomial spread and the initiation of appropriate antimicrobial therapy. The modified Hodge test (MHT), a carbapenem inactivation assay, has shown poor sensitivity in detecting the worldwide spread of New Delhi metallo-β-lactamase (NDM). Recent studies demonstrated that NDM is a lipoprotein anchored to the outer membrane in Gram-negative bacteria, unlike all other known carbapenemases. Here we report that membrane anchoring of β-lactamases precludes detection of carbapenemase activity by the MHT. We also show that this limitation can be overcome by the addition of Triton X-100 during the test, which allows detection of NDM. We propose an improved version of the assay, called the Triton Hodge test (THT), which allows detection of membrane-bound carbapenemases with the addition of this nonionic surfactant. This test was challenged with a panel of 185 clinical isolates (145 carrying known carbapenemase-encoding genes and 40 carbapenemase nonproducers). The THT displayed test sensitivity of >90% against NDM-producing clinical isolates, while improving performance against other carbapenemases. Ertapenem provided the highest sensitivity (97 to 100%, depending on the type of carbapenemase), followed by meropenem (92.5 to 100%). Test specificity was not affected by the addition of Triton (87.5% and 92.5% with ertapenem and meropenem, respectively). This simple inexpensive test confers a large improvement to the sensitivity of the MHT for the detection of NDM and other carbapenemases. PMID:26719442

[Acute meningitis in Córdoba, Colombia (2002-2004)].

PubMed

Tique, Vaneza; Alvis, Nelson; Parodi, Renata; Bustos, Alvaro; Mattar, Salim

2006-05-01

Establishing characteristic epidemiologic and microbiologic features of acute meningitis in the Córdoba department. A descriptive epidemiological study was carried out between June 2002 and June 2004 at the Hospital San Jerónimo in Montería. All suspicious cases of meningitis were included; laboratory tests included cytological smear, biochemistry, latex, Gram stain and culture. 57 (11.3%) and 85 (16.8%) of the 503 samples of cerebrum spinal fluid (CSF) were confirmed by culture as being probable cases. There were 6 cases of polymicrobial infection, making a total of 63 isolates: 17 non-fermenting Gram-negative bacilli (26.9%), 16 Streptococcus pneumoniae (25.4%), 7 Enterobacteriaceae (11%), 5 Criptococcus neoformans (8%) 4 Neisseria meningitidis serotype B (6.3%), 3 S. viridans (4.8%), 2 Streptococcus group B (3.2%), 2 Haemophilus influenzae type B (3.2%), 2 Staphylococcus negative coagulase (3.2%), 2 S. aureus (3.2%), 2 Enterococcus (3.2%) and 1 Candida albicans (1.6%). The S. Pneumoniae serotypes found were: 5 (n=4), 23F (n=3), 14 (n=2), 18C (n=2), 18A (n=l1, 17F (n=l1, 1 (n=1). The study led to determining epidemiological and microbiological aspects of acute meningitis in the Códoba department which had been unknown up to now. Streptococcus pneumoniae (25.4% was the main aetiological agent of meningitis; the epidemiologic aspects so established confirmed the need for strengthening and implementing measures for controlling meningitis in C6ódoba and its surveillance there.

Carbapenem-resistant Gram-negative bacilli in Canada 2009-10: results from the Canadian Nosocomial Infection Surveillance Program (CNISP).

PubMed

Mataseje, L F; Bryce, E; Roscoe, D; Boyd, D A; Embree, J; Gravel, D; Katz, K; Kibsey, P; Kuhn, M; Mounchili, A; Simor, A; Taylor, G; Thomas, E; Turgeon, N; Mulvey, M R

2012-06-01

To investigate the occurrence and molecular mechanisms associated with carbapenemases in carbapenem-resistant Gram-negative isolates from Canadian cases. Twenty hospital sites across Canada submitted isolates for a 1 year period starting 1 September 2009. All Enterobacteriaceae with MICs ≥ 2 mg/L and Acinetobacter baumannii and Pseudomonas aeruginosa with MICs ≥ 16 mg/L of carbapenems were submitted to the National Microbiology Laboratory (NML) where carbapenem MICs were confirmed by Etest and isolates were characterized by PCR for carbapenemase genes, antimicrobial susceptibilities, PFGE and plasmid isolation. A total of 444 isolates (298 P. aeruginosa, 134 Enterobacteriaceae and 12 A. baumannii) were submitted to the NML of which 274 (61.7%; 206 P. aeruginosa, 59 Enterobacteriaceae and 9 A. baumannii) met the inclusion criteria as determined by Etest. Carbapenemase genes were identified in 30 isolates: bla(GES-5) (n = 3; P. aeruginosa), bla(KPC-3) (n = 7; Enterobacteriaceae), bla(NDM-1) (n = 2; Enterobacteriaceae), bla(VIM-2) and bla(VIM-4) (n = 8; P. aeruginosa) bla(SME-2) (n = 1; Enterobacteriaceae) and bla(OXA-23) (n = m9; A. baumannii). PFGE identified a cluster in each of Enterobacteriaceae, P. aeruginosa and A. baumannii corresponding to isolates harbouring carbapenemase genes. Three KPC plasmid patterns (IncN and FllA) were identified where indistinguishable plasmid patterns were identified in unrelated clinical isolates. Carbapenemases were rare at the time of this study. Dissemination of carbapenemases was due to both dominant clones and common plasmid backbones.

Yersinia pestis YopE contains a dominant CD8 T cell epitope that confers protection in a mouse model of pneumonic plague.

PubMed

Lin, Jr-Shiuan; Szaba, Frank M; Kummer, Lawrence W; Chromy, Brett A; Smiley, Stephen T

2011-07-15

Septic bacterial pneumonias are a major cause of death worldwide. Several of the highest priority bioterror concerns, including anthrax, tularemia, and plague, are caused by bacteria that acutely infect the lung. Bacterial resistance to multiple antibiotics is increasingly common. Although vaccines may be our best defense against antibiotic-resistant bacteria, there has been little progress in the development of safe and effective vaccines for pulmonary bacterial pathogens. The Gram-negative bacterium Yersinia pestis causes pneumonic plague, an acutely lethal septic pneumonia. Historic pandemics of plague caused millions of deaths, and the plague bacilli's potential for weaponization sustains an ongoing quest for effective countermeasures. Subunit vaccines have failed, to date, to fully protect nonhuman primates. In mice, they induce the production of Abs that act in concert with type 1 cytokines to deliver high-level protection; however, the Y. pestis Ags recognized by cytokine-producing T cells have yet to be defined. In this study, we report that Y. pestis YopE is a dominant Ag recognized by CD8 T cells in C57BL/6 mice. After vaccinating with live attenuated Y. pestis and challenging intranasally with virulent plague, nearly 20% of pulmonary CD8 T cells recognize this single, highly conserved Ag. Moreover, immunizing mice with a single peptide, YopE(69-77), suffices to confer significant protection from lethal pulmonary challenge. These findings suggest YopE could be a valuable addition to subunit plague vaccines and provide a new animal model in which sensitive, pathogen-specific assays can be used to study CD8 T cell-mediated defense against acutely lethal bacterial infections of the lung.

Epidemiology of infections caused by multiresistant gram-negatives: ESBLs, MBLs, panresistant strains.

PubMed

Rossolini, Gian Maria; Mantengoli, Elisabetta; Docquier, Jean-Denis; Musmanno, Rosa Anna; Coratza, Grazietta

2007-07-01

Microbial drug resistance is a growing problem of global magnitude. In gram-negative pathogens, the most important resistance problems are encountered in Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter, with increasing trends observed for all major anti-gram-negative agents (beta-lactams, fluoroquinolones and aminoglycosides). A matter of major concern is the emergence of new beta-lactamases capable of degrading the expanded-spectrum cephalosporins and/or carbapenems, such as the extended-spectrum beta-lactamases (ESBLs) and the carbapenemases. These beta-lactamase genes are often associated with resistance determinants to non-beta-lactam agents (e.g. aminoglycosides and fluoroquinolones), and strains producing ESBLs or carbapenemases often exhibit complex multidrug resistant phenotypes and sometimes are panresistant. The problem is worsened by the dearth of new agents active on multidrug-resistant Gram-negatives in the pipeline. The importance to develop better strategies to control resistance is underscored.

Enhancing pathogen identification in patients with meningitis and a negative Gram stain using the BioFire FilmArray(®) Meningitis/Encephalitis panel.

PubMed

Wootton, Susan H; Aguilera, Elizabeth; Salazar, Lucrecia; Hemmert, Andrew C; Hasbun, Rodrigo

2016-04-21

Meningitis with a negative cerebrospinal (CSF) Gram stain represents a diagnostic and therapeutic challenge. The purpose of our study was to evaluate the performance of the BioFire FilmArray(®) Meningitis/Encephalitis (FA ME) panel in patients presenting with community-acquired meningitis with a negative Gram stain. CSF from 48 patients with community-acquired meningitis with a negative Gram stain admitted to four hospitals in Houston, TX underwent additional testing by the FA ME. FA ME results were compared to results obtained as part of routine evaluation. The panel detected pathogens not previously identified in 11 (22.9 %) of 48, but did not detect pathogens identified by standard technique (West Nile virus, Histoplasma) in 5 (15.2 %) patients. Rapid testing for the most common pathogens causing meningitis will aid in the diagnosis and treatment of patients with meningitis.

Development and transmission of antimicrobial resistance among Gram-negative bacteria in animals and their public health impact.

PubMed

Mukerji, Shewli; O'Dea, Mark; Barton, Mary; Kirkwood, Roy; Lee, Terence; Abraham, Sam

2017-02-28

Gram-negative bacteria are known to cause severe infections in both humans and animals. Antimicrobial resistance (AMR) in Gram-negative bacteria is a major challenge in the treatment of clinical infections globally due to the propensity of these organisms to rapidly develop resistance against antimicrobials in use. In addition, Gram-negative bacteria possess highly efficient mechanisms through which the AMR can be disseminated between pathogenic and commensal bacteria of the same or different species. These unique traits of Gram-negative bacteria have resulted in evolution of Gram-negative bacterial strains demonstrating resistance to multiple classes of antimicrobials. The evergrowing resistance issue has not only resulted in limitation of treatment options but also led to increased treatment costs and mortality rates in humans and animals. With few or no new antimicrobials in production to combat severe life-threatening infections, AMR has been described as the one of the most severe, long-term threats to human health. Aside from overuse and misuse of antimicrobials in humans, another factor that has exacerbated the emergence of AMR in Gram-negative bacteria is the veterinary use of antimicrobials that belong to the same classes considered to be critically important for treating serious life-threatening infections in humans. Despite the fact that development of AMR dates back to before the introduction of antimicrobials, the recent surge in the resistance towards all available critically important antimicrobials has emerged as a major public health issue. This review thus focuses on discussing the development, transmission and public health impact of AMR in Gram-negative bacteria in animals. © 2017 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Outcomes of single organism peritonitis in peritoneal dialysis: gram negatives versus gram positives in the Network 9 Peritonitis Study.

PubMed

Bunke, C M; Brier, M E; Golper, T A

1997-08-01

The use of the "peritonitis rate" in the management of patients undergoing peritoneal dialysis is assuming importance in comparing the prowess of facilities, care givers and new innovations. For this to be a meaningful outcome measure, the type of infection (causative pathogen) must have less clinical significance than the number of infections during a time interval. The natural history of Staphylococcus aureus, pseudomonas, and fungal peritonitis would not support that the outcome of an episode of peritonitis is independent of the causative pathogen. Could this concern be extended to other more frequently occurring pathogens? To address this, the Network 9 Peritonitis Study identified 530 episodes of single organism peritonitis caused by a gram positive organism and 136 episodes caused by a single non-pseudomonal gram negative (NPGN) pathogen. Coincidental soft tissue infections (exit site or tunnel) occurred equally in both groups. Outcomes of peritonitis were analyzed by organism classification and by presence or absence of a soft tissue infection. NPGN peritonitis was associated with significantly more frequent catheter loss, hospitalization, and technique failure and was less likely to resolve regardless of the presence or absence of a soft tissue infection. Hospitalization and death tended to occur more frequently with enterococcal peritonitis than with other gram positive peritonitis. The outcomes in the NPGN peritonitis group were significantly worse (resolution, catheter loss, hospitalization, technique failure) compared to coagulase negative staphylococcal or S. aureus peritonitis, regardless of the presence or absence of a coincidental soft tissue infection. Furthermore, for the first time, the poor outcomes of gram negative peritonitis are shown to be independent of pseudomonas or polymicrobial involvement or soft tissue infections. The gram negative organism appears to be the important factor. In addition, the outcome of peritonitis caused by S. aureus is worse than that of other staphylococci. Thus, it is clear that all peritonitis episodes cannot be considered equivalent in terms of outcome. The concept of peritonitis rate is only meaningful when specific organisms are considered.

Improved system for floor cleaning in health care facilities.

PubMed Central

Schmidt, E A; Coleman, D L; Mallison, G F

1984-01-01

A new system has been developed for sanitizing floors in hospitals; this system replaces the traditional procedure of daily dusting and wet mopping with a disinfectant-detergent solution and periodic buffing . This new system relies on a sequence of procedures consisting of dust mopping using a chemically treated dust mop, machine buffing of a sprayed-on polymer treatment, and a second dust mopping . The effectiveness of the procedures was evaluated by means of surface sampling for bacterial contamination and air sampling for airborne bacteria and dust. The level of bacterial contamination on the floors was reduced by 93.6% by using the new system, compared with 79.8% by using the conventional process of dust mopping and wet mopping with a disinfectant solution. The levels of airborne bacteria during and after the individual procedures did not vary significantly from the initial level (123.6 CFU/per m3 of air). A survey of representative colonies from air samples revealed staphylococci, gram-positive bacilli, gram-positive diplococci, yeast cells, and infrequent gram-negative rods. The distribution at the conclusion of the sanitizing process was similar to that at the outset. Similarly, the levels of airborne dust measured during and after the individual procedures did not vary significantly from the initial level. When compared with the traditional method of cleaning by wet mopping , the new method was significantly more effective in removal of microbial contamination and required less labor. PMID:6742835

Isolation of Clostridium perfringens type A from wild bharals (Pseudois nayaur) following sudden death in Tibet, China.

PubMed

Zhu, Lingwei; Zhou, Wei; Wang, Tiecheng; Xiang, Haiyang; Ji, Xue; Han, Yixiao; Tian, Yuan; Sun, Yang; Liu, Jun; Guo, Xuejun

2017-04-01

Dozens of wild bharals died suddenly in Tibet. Necropsy showed severe congestion and hemorrhage in multiple organs, with large numbers of Gram-positive bacilli. Strains of Clostridium perfringens type A were isolated from the different organs and the intestinal contents. The other possible pathogens were ruled out by PCR. Copyright © 2017 Elsevier Ltd. All rights reserved.

Female genital tract bacterial coisolates with Candida albicans in patients without clinical vaginitis.

PubMed Central

Monif, G R; Carson, H J

1998-01-01

OBJECTIVE: In vitro, Candida albicans has demonstrated the ability to inhibit replication of selected bacteria. Little information exists on the impact of C. albicans on the vaginal bacterial flora in vivo. The purpose of this study is to identify the coexisting bacterial flora when C. albicans is isolated from vaginal cultures submitted to a hospital-based testing facility for reasons other than vulvovaginitis. METHODOLOGY: All specimens (240) received from ambulatory care clinics over a six-month period were cultured for aerobic and anaerobic bacteria and Candida species. Those specimens submitted for cervicitis, vaginitis, or vaginal discharge and those from which yeasts other than C. albicans were isolated were eliminated. To control for sample biases, a subgroup composed of all pregnant women for whom cultures were done as screening procedures was similarly studied. Chi-square analyses, comparing the prevalence of individual bacteria isolated with and without the presence of C. albicans, were done for all study populations using SPSS for Windows software (1994). RESULTS: Two hundred and forty consecutive specimens were bacteriologically analyzed. Of the 220 vaginal samples used in the study, C. albicans was isolated in 44 instances (20%). Neither the presence of the lactobacilli nor the presence of Gardnerella vaginalis markedly influenced the isolation rate of C. albicans. The group B streptococci had a greater probability of coisolation when C. albicans was present (27.3% verses 16%), but this was not statistically significant (P < 0.8). Dissociation between the presence of C. albicans and the coisolation of Peptostreptococcus species and anaerobic gram-positive cocci and/or bacilli was noted (P < 0.0819), while the incidence of gram-positive aerobic bacilli was reduced in the presence of C. albicans (30/176 [17.1%] versus 6/44 [13.6%]), this reduced incidence was not statistically significant. Isolation data of the subgroup of pregnant women supported these observations. CONCLUSION: Within the limitations of the study, statistically, the data suggests that an inverse relationship exists between the presence of C. albicans and recovery of Peptostreptococcus and anaerobic gram-positive cocci and bacilli. PMID:9702585

Female genital tract bacterial coisolates with Candida albicans in patients without clinical vaginitis.

PubMed

Monif, G R; Carson, H J

1998-01-01

In vitro, Candida albicans has demonstrated the ability to inhibit replication of selected bacteria. Little information exists on the impact of C. albicans on the vaginal bacterial flora in vivo. The purpose of this study is to identify the coexisting bacterial flora when C. albicans is isolated from vaginal cultures submitted to a hospital-based testing facility for reasons other than vulvovaginitis. All specimens (240) received from ambulatory care clinics over a six-month period were cultured for aerobic and anaerobic bacteria and Candida species. Those specimens submitted for cervicitis, vaginitis, or vaginal discharge and those from which yeasts other than C. albicans were isolated were eliminated. To control for sample biases, a subgroup composed of all pregnant women for whom cultures were done as screening procedures was similarly studied. Chi-square analyses, comparing the prevalence of individual bacteria isolated with and without the presence of C. albicans, were done for all study populations using SPSS for Windows software (1994). Two hundred and forty consecutive specimens were bacteriologically analyzed. Of the 220 vaginal samples used in the study, C. albicans was isolated in 44 instances (20%). Neither the presence of the lactobacilli nor the presence of Gardnerella vaginalis markedly influenced the isolation rate of C. albicans. The group B streptococci had a greater probability of coisolation when C. albicans was present (27.3% verses 16%), but this was not statistically significant (P < 0.8). Dissociation between the presence of C. albicans and the coisolation of Peptostreptococcus species and anaerobic gram-positive cocci and/or bacilli was noted (P < 0.0819), while the incidence of gram-positive aerobic bacilli was reduced in the presence of C. albicans (30/176 [17.1%] versus 6/44 [13.6%]), this reduced incidence was not statistically significant. Isolation data of the subgroup of pregnant women supported these observations. Within the limitations of the study, statistically, the data suggests that an inverse relationship exists between the presence of C. albicans and recovery of Peptostreptococcus and anaerobic gram-positive cocci and bacilli.

Epidemiology, susceptibility, and risk factors for acquisition of MDR/XDR Gram-negative bacteria among kidney transplant recipients with urinary tract infections.

PubMed

Yuan, Xiuhong; Liu, Taohua; Wu, Di; Wan, Qiquan

2018-01-01

Multiple drug resistant/extensively drug resistant (MDR/XDR) Gram-negative urinary tract infections (UTIs) represent a growing threat to kidney transplant recipients. This retrospective study aimed to assess the incidence and microbiological profile of MDR/XDR Gram-negative UTIs, to identify drug susceptibility of MDR/XDR bacteria, and to determine the potential risk factors for MDR/XDR UTIs in kidney recipients. During the study period, 1569 patients underwent consecutive kidney transplantation in two transplantation centers. We studied the demographics, clinical characteristics, and urine culture data from kidney recipients with MDR/XDR Gram-negative UTIs, and verified the risk factors associated with MDR/XDR infections. Eighty-one kidney recipients yielded 88 episodes of MDR/XDR Gram-negative UTIs with five patients (6.2%) succumbing to all-cause in-hospital mortality. The most frequently isolated bacterium was Escherichia coli (62.5%). Almost all MDR/XDR Gram-negative bacteria were resistant to first- and second-generation cephalosporin, and monocyclic beta-lactam. They were relatively sensitive to meropenem, amikacin, and tigecycline. As for the 12 XDR bacteria, all of them were resistant to meropenem and 25% of them were resistant to tigecycline. All XDR Acinetobacter baumannii and E. coli were susceptible to tigecycline. Nosocomial infection (odds ratio [OR] = 11.429, 95% CI = 1.311-99.625, P = 0.027) was the only independent predictor of MDR/XDR Gram-negative UTIs. Non-fermenting bacterial infection (OR = 20.161, 95% CI = 3.409-119.240, P = 0.001), polycystic kidney disease (OR = 39.871, 95% CI = 1.979-803.384, P = 0.016), and serum creatinine level > 1.5 mg/dL (OR = 8.688, 95% CI = 1.354-55.747, P = 0.023) were significantly different between XDR and MDR Gram-negative UTIs. Meropenem, amikacin, and/or tigecycline can be prescribed for MDR/XDR Gram-negative infections. Tigecycline can also be prescribed for XDR A. baumannii and E. coli . Nosocomial infection was a risk factor for MDR/XDR Gram-negative UTIs, while XDR UTIs were associated with non-fermenting bacterial infection, polycystic kidney disease, and impaired renal function.

Bactericidal catechins damage the lipid bilayer.

PubMed

Ikigai, H; Nakae, T; Hara, Y; Shimamura, T

1993-04-08

The mode of antibacterial action of, the green tea (Camellia sinensis) extracts, (-)-epigallocatechin gallate (EGCg) and (-)-epicatechin (EC) was investigated. Strong bactericidal EGCg caused leakage of 5,6-carboxyfluorescein from phosphatidylcholine liposomes (PC), but EC with very weak bactericidal activity caused little damage to the membrane. Phosphatidylserine and dicetyl phosphate partially protected the membrane from EGCg-mediated damage when reconstituted into the liposome membrane with PC. EGCg, but not EC, caused strong aggregation and NPN-fluorescence quenching of PC-liposomes and these actions were markedly lowered in the presence of negatively charged lipids. These results show that bactericidal catechins primarily act on and damage bacterial membranes. The observation that Gram-negative bacteria are more resistant to bactericidal catechins than Gram-positive bacteria can be explained to some extent by the presence of negatively charged lipopolysaccharide.

Automated detection of tuberculosis on sputum smeared slides using stepwise classification

NASA Astrophysics Data System (ADS)

Divekar, Ajay; Pangilinan, Corina; Coetzee, Gerrit; Sondh, Tarlochan; Lure, Fleming Y. M.; Kennedy, Sean

2012-03-01

Routine visual slide screening for identification of tuberculosis (TB) bacilli in stained sputum slides under microscope system is a tedious labor-intensive task and can miss up to 50% of TB. Based on the Shannon cofactor expansion on Boolean function for classification, a stepwise classification (SWC) algorithm is developed to remove different types of false positives, one type at a time, and to increase the detection of TB bacilli at different concentrations. Both bacilli and non-bacilli objects are first analyzed and classified into several different categories including scanty positive, high concentration positive, and several non-bacilli categories: small bright objects, beaded, dim elongated objects, etc. The morphological and contrast features are extracted based on aprior clinical knowledge. The SWC is composed of several individual classifiers. Individual classifier to increase the bacilli counts utilizes an adaptive algorithm based on a microbiologist's statistical heuristic decision process. Individual classifier to reduce false positive is developed through minimization from a binary decision tree to classify different types of true and false positive based on feature vectors. Finally, the detection algorithm is was tested on 102 independent confirmed negative and 74 positive cases. A multi-class task analysis shows high accordance rate for negative, scanty, and high-concentration as 88.24%, 56.00%, and 97.96%, respectively. A binary-class task analysis using a receiver operating characteristics method with the area under the curve (Az) is also utilized to analyze the performance of this detection algorithm, showing the superior detection performance on the high-concentration cases (Az=0.913) and cases mixed with high-concentration and scanty cases (Az=0.878).

Sequence- and structure-based computational analyses of Gram-negative tripartite efflux pumps in the context of bacterial membranes

DOE PAGES

Travers, Timothy; Wang, Katherine J.; Lopez, Cesar A.; ...

2018-02-09

Gram-negative multidrug resistance currently presents a serious threat to public health with infections effectively rendered untreatable. Multiple molecular mechanisms exist that cause antibiotic resistance and in addition, the last three decades have seen slowing rates of new drug development. In this paper, we summarize the use of various computational techniques for investigating the mechanisms of multidrug resistance mediated by Gram-negative tripartite efflux pumps and membranes. Recent work in our lab combines data-driven sequence and structure analyses to study the interactions and dynamics of these bacterial components. Computational studies can complement experimental methodologies for gaining crucial insights into combatting multidrug resistance.

Sequence- and structure-based computational analyses of Gram-negative tripartite efflux pumps in the context of bacterial membranes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Travers, Timothy; Wang, Katherine J.; Lopez, Cesar A.

Gram-negative multidrug resistance currently presents a serious threat to public health with infections effectively rendered untreatable. Multiple molecular mechanisms exist that cause antibiotic resistance and in addition, the last three decades have seen slowing rates of new drug development. In this paper, we summarize the use of various computational techniques for investigating the mechanisms of multidrug resistance mediated by Gram-negative tripartite efflux pumps and membranes. Recent work in our lab combines data-driven sequence and structure analyses to study the interactions and dynamics of these bacterial components. Computational studies can complement experimental methodologies for gaining crucial insights into combatting multidrug resistance.

A Product of Heme Catabolism Modulates Bacterial Function and Survival

PubMed Central

Nobles, Christopher L.; Green, Sabrina I.; Maresso, Anthony W.

2013-01-01

Bilirubin is the terminal metabolite in heme catabolism in mammals. After deposition into bile, bilirubin is released in large quantities into the mammalian gastrointestinal (GI) tract. We hypothesized that intestinal bilirubin may modulate the function of enteric bacteria. To test this hypothesis, we investigated the effect of bilirubin on two enteric pathogens; enterohemorrhagic E. coli (EHEC), a Gram-negative that causes life-threatening intestinal infections, and E. faecalis, a Gram-positive human commensal bacterium known to be an opportunistic pathogen with broad-spectrum antibiotic resistance. We demonstrate that bilirubin can protect EHEC from exogenous and host-generated reactive oxygen species (ROS) through the absorption of free radicals. In contrast, E. faecalis was highly susceptible to bilirubin, which causes significant membrane disruption and uncoupling of respiratory metabolism in this bacterium. Interestingly, similar results were observed for other Gram-positive bacteria, including B. cereus and S. aureus. A model is proposed whereby bilirubin places distinct selective pressure on enteric bacteria, with Gram-negative bacteria being protected from ROS (positive outcome) and Gram-positive bacteria being susceptible to membrane disruption (negative outcome). This work suggests bilirubin has differential but biologically relevant effects on bacteria and justifies additional efforts to determine the role of this neglected waste catabolite in disease processes, including animal models. PMID:23935485

Membrane-active macromolecules kill antibiotic-tolerant bacteria and potentiate antibiotics towards Gram-negative bacteria

PubMed Central

Uppu, Divakara S. S. M.; Konai, Mohini M.; Sarkar, Paramita; Samaddar, Sandip; Fensterseifer, Isabel C. M.; Farias-Junior, Celio; Krishnamoorthy, Paramanandam; Shome, Bibek R.; Franco, Octávio L.

2017-01-01

Chronic bacterial biofilms place a massive burden on healthcare due to the presence of antibiotic-tolerant dormant bacteria. Some of the conventional antibiotics such as erythromycin, vancomycin, linezolid, rifampicin etc. are inherently ineffective against Gram-negative bacteria, particularly in their biofilms. Here, we report membrane-active macromolecules that kill slow dividing stationary-phase and antibiotic tolerant cells of Gram-negative bacteria. More importantly, these molecules potentiate antibiotics (erythromycin and rifampicin) to biofilms of Gram-negative bacteria. These molecules eliminate planktonic bacteria that are liberated after dispersion of biofilms (dispersed cells). The membrane-active mechanism of these molecules forms the key for potentiating the established antibiotics. Further, we demonstrate that the combination of macromolecules and antibiotics significantly reduces bacterial burden in mouse burn and surgical wound infection models caused by Acinetobacter baumannii and Carbapenemase producing Klebsiella pneumoniae (KPC) clinical isolate respectively. Colistin, a well-known antibiotic targeting the lipopolysaccharide (LPS) of Gram-negative bacteria fails to kill antibiotic tolerant cells and dispersed cells (from biofilms) and bacteria develop resistance to it. On the contrary, these macromolecules prevent or delay the development of bacterial resistance to known antibiotics. Our findings emphasize the potential of targeting the bacterial membrane in antibiotic potentiation for disruption of biofilms and suggest a promising strategy towards developing therapies for topical treatment of Gram-negative infections. PMID:28837596

Executive summary: Diagnosis and Treatment of Catheter-Related Bloodstream Infection: Clinical Guidelines of the Spanish Society of Clinical Microbiology and Infectious Diseases (SEIMC) and the Spanish Society of Intensive Care Medicine and Coronary Units (SEMICYUC).

PubMed

Chaves, Fernando; Garnacho-Montero, José; Del Pozo, José Luis; Bouza, Emilio; Capdevila, José Antonio; de Cueto, Marina; Domínguez, M Ángeles; Esteban, Jaime; Fernández-Hidalgo, Nuria; Fernández Sampedro, Marta; Fortún, Jesús; Guembe, María; Lorente, Leonardo; Paño, Jose Ramón; Ramírez, Paula; Salavert, Miguel; Sánchez, Miguel; Vallés, Jordi

2018-02-01

Catheter-related bloodstream infections (CRBSI) constitute an important cause of hospital-acquired infection associated with morbidity, mortality, and cost. The aim of these guidelines is to provide updated recommendations for the diagnosis and management of CRBSI in adults. Prevention of CRBSI is excluded. Experts in the field were designated by the two participating Societies (Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica and the Sociedad Española de Medicina Intensiva, Crítica y Unidades Coronarias). Short-term peripheral venous catheters, non-tunneled and long-term central venous catheters, tunneled catheters and hemodialysis catheters are covered by these guidelines. The panel identified 39 key topics that were formulated in accordance with the PICO format. The strength of the recommendations and quality of the evidence were graded in accordance with ESCMID guidelines. Recommendations are made for the diagnosis of CRBSI with and without catheter removal and of tunnel infection. The document establishes the clinical situations in which a conservative diagnosis of CRBSI (diagnosis without catheter removal) is feasible. Recommendations are also made regarding empirical therapy, pathogen-specific treatment (coagulase-negative staphylococci, Sthaphylococcus aureus, Enterococcus spp, Gram-negative bacilli, and Candida spp), antibiotic lock therapy, diagnosis and management of suppurative thrombophlebitis and local complications. Copyright © 2017 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

Isolation and analysis of bacteria with antimicrobial activities from the marine sponge Haliclona simulans collected from Irish waters.

PubMed

Kennedy, Jonathan; Baker, Paul; Piper, Clare; Cotter, Paul D; Walsh, Marcella; Mooij, Marlies J; Bourke, Marie B; Rea, Mary C; O'Connor, Paula M; Ross, R Paul; Hill, Colin; O'Gara, Fergal; Marchesi, Julian R; Dobson, Alan D W

2009-01-01

Samples of the marine sponge Haliclona simulans were collected from Irish coastal waters, and bacteria were isolated from these samples. Phylogenetic analyses of the cultured isolates showed that four different bacterial phyla were represented; Bacteriodetes, Actinobacteria, Proteobacteria, and Firmicutes. The sponge bacterial isolates were assayed for the production of antimicrobial substances, and biological activities against Gram-positive and Gram-negative bacteria and fungi were demonstrated, with 50% of isolates showing antimicrobial activity against at least one of the test strains. Further testing showed that the antimicrobial activities extended to the important pathogens Pseudomonas aeruginosa, Clostridium difficile, multi-drug-resistant Staphylococcus aureus, and pathogenic yeast strains. The Actinomycetes were numerically the most abundant producers of antimicrobial activities, although activities were also noted from Bacilli and Pseudovibrio isolates. Surveys for the presence of potential antibiotic encoding polyketide synthase and nonribosomal peptide synthetase genes also revealed that genes for the biosynthesis of these secondary metabolites were present in most bacterial phyla but were particularly prevalent among the Actinobacteria and Proteobacteria. This study demonstrates that the culturable fraction of bacteria from the sponge H. simulans is diverse and appears to possess much potential as a source for the discovery of new medically relevant biological active agents.

Microbiological characterization of the biological treatment of aircraft paint stripping wastewater.

PubMed

Arquiaga, M C; Canter, L W; Robertson, J M

1995-01-01

Research on the treatment of potentially toxic wastewater produced at six US Navy aircraft paint stripping facilities has been conducted. The composition of the wastewater treated consisted of methylene chloride and phenol in concentrations of about 5000 and 1800 mg/l, respectively, and other organic compounds in a total concentration of 2200 mg/l. Biological treatment is an important means by which toxic or hazardous organic compounds can be economically converted to less noxious materials. Engineering studies conducted in the laboratory with activated sludge reactors and rotating biological contactors (RBC) demonstrated that both suspended and attached growths can be effective biological methods to treat this paint stripping wastewater when blended with domestic wastewater up to about 50% by volume. These studies were complemented with analyses of the bacterial communities inhabiting the treatment systems. The number and the genera of the microorganisms present in the blended wastewater, as well as their ability to biodegrade the potentially toxic organics were studied. The results indicate that paint stripping wastewater is able to support large bacterial populations consisting of various gram-negative rods and coccibacilli and a few gram-positive bacilli. Members of the genera Pseudomonas and Bacillus are suspected to play an important role in initiating the biodegradation process.

[Multidrug resistance E-ESKAPE strains isolated from blood cultures in patients with cancer].

PubMed

Velázquez-Acosta, Consuelo; Cornejo-Juárez, Patricia; Volkow-Fernández, Patricia

2018-01-01

To describe the trend of multidrug resistant (MDR) strains isolated from blood in patients with cancer from 2005 to 2015. 33 127 blood cultures were processed by retrospective analysis. Identification and antimicrobial sensitivity were performed through automated methods: WaLK away (Siemens Labora- tory Diagnostics) and BD Phoenix (Becton, Dickinson and Company). Resistant strains were determined according to the minimum inhibitory concentration, following the parameters of the Clinical and Laboratory Standards Institute (CLSI). Of 6 397 isolates, 5 604 (16.9%) were positive; 3 732 (58.4%) Gram- bacilli; 2 355 (36.9%) Gram+ cocci; 179 (2.7%) yeasts, and 126 (1.9%) Gram+ bacilli. Escherichia coli (n=1 591, 24.5%) was the most frequent bacteria, with 652 (41%) strains being extended-spectrum beta-lactamases producers (ESBL); of Enterococcus faecium (n=143, 2.1%), 45 (31.5%) were vancomycin resistant; of Staphylococcus aureus (n=571, 8.7%), 121 (21.2%) methicillin resistant (MRSA); of Klebsiella pneumoniae (n=367, 5.6%), 41 (11.2%) ESBL; of Acinetobacter baumanii (n=96, 1.4%), 23 (24%) MDR, and of Pseudomonas aeruginosa (n=384, 5.6%), 43 (11.2%) MDR. MDR strains were significantly more frequent in patients with hematological malignancies, compared to those with solid tumors: MRSA (OR=4.48, 95%CI 2.9-6.8), ESBL E. coli(OR=1.3, 95%CI 1.10-1.65) and MDR Acinetobacter baumanii (OR=3.2, 95%CI 1.2-8.3). We observed significantly higher isolations of E-ESPAKE MDR strains in patients with hematological malignancies.

Genomic determinants of sporulation in Bacilli and Clostridia: towards the minimal set of sporulation-specific genes

PubMed Central

Galperin, Michael Y; Mekhedov, Sergei L; Puigbo, Pere; Smirnov, Sergey; Wolf, Yuri I; Rigden, Daniel J

2012-01-01

Three classes of low-G+C Gram-positive bacteria (Firmicutes), Bacilli, Clostridia and Negativicutes, include numerous members that are capable of producing heat-resistant endospores. Spore-forming firmicutes include many environmentally important organisms, such as insect pathogens and cellulose-degrading industrial strains, as well as human pathogens responsible for such diseases as anthrax, botulism, gas gangrene and tetanus. In the best-studied model organism Bacillus subtilis, sporulation involves over 500 genes, many of which are conserved among other bacilli and clostridia. This work aimed to define the genomic requirements for sporulation through an analysis of the presence of sporulation genes in various firmicutes, including those with smaller genomes than B. subtilis. Cultivable spore-formers were found to have genomes larger than 2300 kb and encompass over 2150 protein-coding genes of which 60 are orthologues of genes that are apparently essential for sporulation in B. subtilis. Clostridial spore-formers lack, among others, spoIIB, sda, spoVID and safA genes and have non-orthologous displacements of spoIIQ and spoIVFA, suggesting substantial differences between bacilli and clostridia in the engulfment and spore coat formation steps. Many B. subtilis sporulation genes, particularly those encoding small acid-soluble spore proteins and spore coat proteins, were found only in the family Bacillaceae, or even in a subset of Bacillus spp. Phylogenetic profiles of sporulation genes, compiled in this work, confirm the presence of a common sporulation gene core, but also illuminate the diversity of the sporulation processes within various lineages. These profiles should help further experimental studies of uncharacterized widespread sporulation genes, which would ultimately allow delineation of the minimal set(s) of sporulation-specific genes in Bacilli and Clostridia. PMID:22882546

Chlamydia pneumoniae Infection Among Basic Underwater Demolition/SEAL (BUD/S) Trainees, Coronado, California, July 2008

DTIC Science & Technology

2011-03-01

including Streptococcus pneumoniae, Chlamydophila pneu- moniae, Mycoplasma pneumoniae, Legionella pneumophila, and Bordetella pertussis . 3 CAP...CAP. 15 Additionally, radiological fi ndings have a limited capacity to differentiate the etiologic agents of CAP. 16 Because treatment modalities...and gram-positive bacilli and have been used in the treatment of atyp- ical pneumonias including M pneumoniae and C pneumoniae . 26 The need to

Dialysate bacterial endotoxin as a prognostic indicator of peritoneal dialysis related peritonitis.

PubMed

Szeto, Cheuk-Chun; Lai, Ka-Bik; Chow, Kai-Ming; Kwan, Bonnie Ching-Ha; Law, Man-Ching; Pang, Wing-Fai; Ma, Terry King-Wing; Leung, Chi-Bon; Li, Philip Kam-Tao

2016-12-01

Peritonitis is the major complication of peritoneal dialysis (PD). The aim of our present study is to explore the prognostic value of endotoxin level in PD effluent for the prediction of treatment failure in PD-related peritonitis. We studied 325 peritonitis episodes in 223 patients. PD effluent (PDE) was collected every 5 days for endotoxin level and leukocyte count. Patients were followed for relapsing or recurrent peritonitis. We found 20 episodes (6.2%) had primary treatment failure; 41 (12.6%) developed relapsing, 19 (5.8%) had recurrent, and 22 (6.8%) had repeat episodes. Endotoxin was detectable in the PDE of 19 episodes (24.4%) caused by Gram negative organisms, 4 episodes (6.8%) of mixed bacterial growth, and none of the culture negative episodes or those by Gram positive organisms. For episodes caused by Gram negative bacteria, a detectable endotoxin level in PDE on day 5 had a sensitivity and specificity of 66.7% and 83.3%, respectively, for predicting primary treatment failure. In contrast, PDE leukocyte count > 1000 per mm3 on day 5 had a sensitivity and specificity of 88.9% and 89.1%, respectively; the addition of PDE endotoxin assay did not improve the sensitivity or specificity. We conclude that detectable endotoxin in PDE 5 days after antibiotic therapy might predict primary treatment failure in peritonitis episodes caused by Gram negative organisms. However, the sensitivity and specificity of PDE endotoxin assay was inferior to PDE leukocyte count. © 2016 Asian Pacific Society of Nephrology.

A Novel Enzyme-Linked Immunosorbent Assay for Diagnosis of Mycobacterium avium subsp. paratuberculosis Infections (Johne's Disease) in Cattle

PubMed Central

Speer, C. A.; Scott, M. Cathy; Bannantine, John P.; Waters, W. Ray; Mori, Yasuyuki; Whitlock, Robert H.; Eda, Shigetoshi

2006-01-01

Enzyme-linked immunosorbent assays (ELISAs) for the diagnosis of Johne's disease (JD), caused by Mycobacterium avium subsp. paratuberculosis, were developed using whole bacilli treated with formaldehyde (called WELISA) or surface antigens obtained by treatment of M. avium subsp. paratuberculosis bacilli with formaldehyde and then brief sonication (called SELISA). ELISA plates were coated with either whole bacilli or sonicated antigens and tested for reactivity against serum obtained from JD-positive and JD-negative cattle or from calves experimentally inoculated with M. avium subsp. paratuberculosis, Mycobacterium avium subsp. avium, or Mycobacterium bovis. Because the initial results obtained from the WELISA and SELISA were similar, most of the subsequent experiments reported herein were performed using the SELISA method. To optimize the SELISA test, various concentrations (3.7 to 37%) of formaldehyde and intervals of sonication (2 to 300 s) were tested. With an increase in formaldehyde concentration and a decreased interval of sonication, there was a concomitant decrease in nonspecific binding by the SELISA. SELISAs prepared by treating M. avium subsp. paratuberculosis with 37% formaldehyde and then a 2-s burst of sonication produced the greatest difference (7×) between M. avium subsp. paratuberculosis-negative and M. avium subsp. paratuberculosis-positive serum samples. The diagnostic sensitivity and specificity for JD by the SELISA were greater than 95%. The SELISA showed subspecies-specific detection of M. avium subsp. paratuberculosis infections in calves experimentally inoculated with M. avium subsp. paratuberculosis or other mycobacteria. Based on diagnostic sensitivity and specificity, the SELISA appears superior to the commercial ELISAs routinely used for the diagnosis of JD. PMID:16682472

Burn-associated bloodstream infections in pediatric burn patients: Time distribution of etiologic agents.

PubMed

Devrim, İlker; Kara, Ahu; Düzgöl, Mine; Karkıner, Aytaç; Bayram, Nuri; Temir, Günyüz; Şencan, Arzu; Sorguç, Yelda; Gülfidan, Gamze; Hoşgör, Münevver

2017-02-01

Infections are the leading cause of morbidity and mortality in patients with burns in burn units. Bloodstream infections (BSIs) in patients with burns may result from burn wound infection, use of invasive devices such as central venous catheters, and translocation of the gastrointestinal flora. In this study, we investigated the distribution and antimicrobial drug resistance of causative pathogens in children with burns and the durational changes of microorganisms in the distribution of BSIs in children. This study was conducted at the Pediatric Burn Unit (PBU) of Dr. Behçet Uz Children Research and Training Hospital during the period of November 2008-April 2015. The study subjects were all the patients admitted to the PBU, in whom microorganisms were isolated at least from one of the cultures, including blood and catheter cultures. Gram-positive bacteria were the most common causative agents of BSI in patients with burns (66.4%), followed by gram-negative bacteria (22.1%) and fungi (11.5%). The median duration of development of BSIs caused by gram-positive bacteria from the time of burn was 5 days (ranging from 2 to 54 days of burn), which was significantly shorter than that of BSIs caused by gram-negative bacteria (12 days) and fungal pathogens (13 days). The etiologic agents of BSIs in children may differ from those in adults. Gram-negative drug-resistant bacteria such as multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii were important agents of BSI in patients with burns, especially in the long term; however, gram-positive bacteria should also be considered while deciding the antimicrobial therapy, especially in the early periods of burn. Copyright © 2016 Elsevier Ltd and ISBI. All rights reserved.

Postoperative infection in spine surgery: does the month matter?

PubMed

Durkin, Michael J; Dicks, Kristen V; Baker, Arthur W; Moehring, Rebekah W; Chen, Luke F; Sexton, Daniel J; Lewis, Sarah S; Anderson, Deverick J

2015-07-01

The relationship between time of year and surgical site infection (SSI) following neurosurgical procedures is poorly understood. Authors of previous reports have demonstrated that rates of SSI following neurosurgical procedures performed during the summer months were higher compared with rates during other seasons. It is unclear, however, if this difference was related to climatological changes or inexperienced medical trainees (the July effect). The aim of this study was to evaluate for seasonal variation of SSI following spine surgery in a network of nonteaching community hospitals. The authors analyzed 6 years of prospectively collected surveillance data (January 1, 2007, to December 31, 2012) from all laminectomies and spinal fusions from 20 hospitals in the Duke Infection Control Outreach Network of community hospitals. Surgical site infections were defined using National Healthcare Safety Network criteria and identified using standardized methods across study hospitals. Regression models were then constructed using Poisson distribution to evaluate for seasonal trends by month. Each analysis was first performed for all SSIs and then for SSIs caused by specific organisms or classes of organisms. Categorical analysis was performed using two separate definitions of summer: June through September (definition 1), and July through September (definition 2). The prevalence rate of SSIs during the summer was compared with the prevalence rate during the remainder of the year by calculating prevalence rate ratios and 95% confidence intervals. The authors identified 642 SSIs following 57,559 neurosurgical procedures (overall prevalence rate = 1.11/100 procedures); 215 occurred following 24,466 laminectomies (prevalence rate = 0.88/100 procedures), and 427 following 33,093 spinal fusions (prevalence rate = 1.29/100 procedures). Common causes of SSI were Staphylococcus aureus (n = 380; 59%), coagulase-negative staphylococci (n = 90; 14%), and Escherichia coli (n = 41; 6.4%). Poisson regression models demonstrated increases in the rates of SSI during each of the summer months for all SSIs and SSIs due to gram-positive cocci, S. aureus, and methicillin-sensitive S. aureus. Categorical analysis confirmed that the rate of SSI during the 4-month summer period was higher than the rate during the remainder of the year, regardless of which definition for summer was used (definition 1, p = 0.008; definition 2, p = 0.003). Similarly, the rates of SSI due to grampositive cocci and S. aureus were higher during the summer months than the remainder of the year regardless of which definition of summer was used. However, the rate of SSI due to gram-negative bacilli was not. The rate of SSI following fusion or spinal laminectomy/laminoplasty was higher during the summer in this network of community hospitals. The increase appears to be related to increases in SSIs caused by gram-positive cocci and, more specifically, S. aureus. Given the nonteaching nature of these hospitals, the findings demonstrate that increases in the rate of SSI during the summer are more likely related to ecological and/or environmental factors than the July effect.

Bloodstream Infections in Patients With Solid Tumors

PubMed Central

Marín, Mar; Gudiol, Carlota; Garcia-Vidal, Carol; Ardanuy, Carmen; Carratalà, Jordi

2014-01-01

Abstract Current information regarding bloodstream infection (BSI) in patients with solid tumors is scarce. We assessed the epidemiology, antibiotic therapy, and outcomes of BSI in these patients. We also compared patients who died with those who survived to identify risk factors associated with mortality. From January 2006 to July 2012 all episodes of BSI in patients with solid tumors at a cancer center were prospectively recorded and analyzed. A total of 528 episodes of BSI were documented in 489 patients. The most frequent neoplasms were hepatobiliary tumors (19%), followed by lung cancer (18%) and lower gastrointestinal malignancies (16%). Many patients had received corticosteroid therapy (41%), and 15% had neutropenia (<500 neutrophils/μL) at the time of BSI. The most common source of BSI was cholangitis (21%), followed by other abdominal (19.5%) and urinary tract infections (17%). Gram-negative BSI occurred in 55% of cases, mainly due to Escherichia coli (55%), Pseudomonas aeruginosa (18%), and Klebsiella pneumoniae (16%). Among gram-positive BSI (35%), viridans group streptococci were the most frequent causative organisms (22%), followed by Staphylococcus aureus (21%) and Enterococcus species (18%). We identified 61 multidrug-resistant (MDR) organisms (13%), mainly extended-spectrum β-lactamase-producing Enterobacteriaceae (n = 20) and AmpC-producing Enterobacteriaceae (n = 13). The majority of patients with BSI caused by MDR organisms had received antibiotics (70%), and they had been previously hospitalized (61.4%) more frequently than patients with BSI caused by susceptible strains. Inadequate empirical antibiotic therapy was given to 23% of patients, with a higher proportion in those with BSI due to a MDR strain (69%). Early (<48 h) and overall (30 d) case-fatality rates were 7% and 32%, respectively. The overall case-fatality rate was higher among cases caused by MDR organisms (39.3%). The only independent risk factors for the early case-fatality rate were the endogenous source of BSI (odds ratio [OR], 3.57; 95% confidence interval [CI], 1.06–12.02), shock at presentation (OR, 3.63; 95% CI, 1.63–8.09), and corticosteroid therapy (OR, 3.245; 95% CI, 1.43–7.32). The independent risk factors for overall case-fatality rate were the presence of a chronic advanced cancer (OR, 35.39; 95% CI, 2.48–504.91), shock at presentation (OR, 25.84; 95% CI, 3.73–179.0), and corticosteroid therapy (OR, 6.98; 95% CI, 1.61–30.21). BSI in patients with solid tumors occurred mainly among those with hepatobiliary cancer, and cholangitis was the most frequent source; gram-negative bacilli were the most frequent causative agents. MDR organisms were relatively common, particularly in patients who had previously received antibiotics and had been hospitalized; these patients were frequently treated with inadequate empirical antibiotic therapy and had a poorer outcome. The case-fatality rate of patients with solid tumors and BSI was high and was associated with chronic advanced cancer, corticosteroid therapy, and shock at presentation. PMID:24797169

Bloodstream infections in patients with solid tumors: epidemiology, antibiotic therapy, and outcomes in 528 episodes in a single cancer center.

PubMed

Marín, Mar; Gudiol, Carlota; Garcia-Vidal, Carol; Ardanuy, Carmen; Carratalà, Jordi

2014-05-01

Current information regarding bloodstream infection (BSI) in patients with solid tumors is scarce. We assessed the epidemiology, antibiotic therapy, and outcomes of BSI in these patients. We also compared patients who died with those who survived to identify risk factors associated with mortality. From January 2006 to July 2012 all episodes of BSI in patients with solid tumors at a cancer center were prospectively recorded and analyzed. A total of 528 episodes of BSI were documented in 489 patients. The most frequent neoplasms were hepatobiliary tumors (19%), followed by lung cancer (18%) and lower gastrointestinal malignancies (16%). Many patients had received corticosteroid therapy (41%), and 15% had neutropenia (<500 neutrophils/μL) at the time of BSI. The most common source of BSI was cholangitis (21%), followed by other abdominal (19.5%) and urinary tract infections (17%). Gram-negative BSI occurred in 55% of cases, mainly due to Escherichia coli (55%), Pseudomonas aeruginosa (18%), and Klebsiella pneumoniae (16%). Among gram-positive BSI (35%), viridans group streptococci were the most frequent causative organisms (22%), followed by Staphylococcus aureus (21%) and Enterococcus species (18%). We identified 61 multidrug-resistant (MDR) organisms (13%), mainly extended-spectrum β-lactamase-producing Enterobacteriaceae (n = 20) and AmpC-producing Enterobacteriaceae (n = 13). The majority of patients with BSI caused by MDR organisms had received antibiotics (70%), and they had been previously hospitalized (61.4%) more frequently than patients with BSI caused by susceptible strains. Inadequate empirical antibiotic therapy was given to 23% of patients, with a higher proportion in those with BSI due to a MDR strain (69%). Early (<48 h) and overall (30 d) case-fatality rates were 7% and 32%, respectively. The overall case-fatality rate was higher among cases caused by MDR organisms (39.3%). The only independent risk factors for the early case-fatality rate were the endogenous source of BSI (odds ratio [OR], 3.57; 95% confidence interval [CI], 1.06-12.02), shock at presentation (OR, 3.63; 95% CI, 1.63-8.09), and corticosteroid therapy (OR, 3.245; 95% CI, 1.43-7.32). The independent risk factors for overall case-fatality rate were the presence of a chronic advanced cancer (OR, 35.39; 95% CI, 2.48-504.91), shock at presentation (OR, 25.84; 95% CI, 3.73-179.0), and corticosteroid therapy (OR, 6.98; 95% CI, 1.61-30.21).BSI in patients with solid tumors occurred mainly among those with hepatobiliary cancer, and cholangitis was the most frequent source; gram-negative bacilli were the most frequent causative agents. MDR organisms were relatively common, particularly in patients who had previously received antibiotics and had been hospitalized; these patients were frequently treated with inadequate empirical antibiotic therapy and had a poorer outcome. The case-fatality rate of patients with solid tumors and BSI was high and was associated with chronic advanced cancer, corticosteroid therapy, and shock at presentation.

The Growing Threat of Multidrug-Resistant Gram-Negative Infections in Patients with Hematologic Malignancies

PubMed Central

Baker, Thomas M.; Satlin, Michael J.

2016-01-01

Prolonged neutropenia and chemotherapy-induced mucositis render patients with hematologic malignancies highly vulnerable to Gram-negative bacteremia. Unfortunately, multidrug-resistant (MDR) Gram-negative bacteria are increasingly encountered globally, and current guidelines for empirical antibiotic coverage in these patients may not adequately treat these bacteria. This expansion of resistance, coupled with traditional culturing techniques requiring 2-4 days for bacterial identification and antimicrobial susceptibility results, have grave implications for these immunocompromised hosts. This review characterizes the epidemiology, risk factors, resistance mechanisms, recommended treatments, and outcomes of the MDR Gram-negative bacteria that commonly cause infections in patients with hematologic malignancies. We also examine infection prevention strategies in hematology patients, such as infection control practices, antimicrobial stewardship, and targeted decolonization. Finally, we assess strategies to improve outcomes of infected patients, including gastrointestinal screening to guide empirical antibiotic therapy, new rapid diagnostic tools for expeditious identification of MDR pathogens, and use of two new antimicrobial agents, ceftolozane/tazobactam and ceftazidime/avibactam. PMID:27339405

Antibiotic resistance: What is so special about multidrug-resistant Gram-negative bacteria?

PubMed Central

Exner, Martin; Bhattacharya, Sanjay; Christiansen, Bärbel; Gebel, Jürgen; Goroncy-Bermes, Peter; Hartemann, Philippe; Heeg, Peter; Ilschner, Carola; Kramer, Axel; Larson, Elaine; Merkens, Wolfgang; Mielke, Martin; Oltmanns, Peter; Ross, Birgit; Rotter, Manfred; Schmithausen, Ricarda Maria; Sonntag, Hans-Günther; Trautmann, Matthias

2017-01-01

In the past years infections caused by multidrug-resistant Gram-negative bacteria have dramatically increased in all parts of the world. This consensus paper is based on presentations, subsequent discussions and an appraisal of current literature by a panel of international experts invited by the Rudolf Schülke Stiftung, Hamburg. It deals with the epidemiology and the inherent properties of Gram-negative bacteria, elucidating the patterns of the spread of antibiotic resistance, highlighting reservoirs as well as transmission pathways and risk factors for infection, mortality, treatment and prevention options as well as the consequences of their prevalence in livestock. Following a global, One Health approach and based on the evaluation of the existing knowledge about these pathogens, this paper gives recommendations for prevention and infection control measures as well as proposals for various target groups to tackle the threats posed by Gram-negative bacteria and prevent the spread and emergence of new antibiotic resistances. PMID:28451516

In vitro growth inhibition of mastitis causing bacteria by phenolics and metal chelators

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chew, B.P.; Tjoelker, L.W.; Tanaka, T.S.

1985-11-01

Antimicrobial activities of three phenolic compounds and four metal chelators were tested at 0, 250, 500, and 1000 ppm in vitro against four major mastitis-causing bacteria, Streptococcus agalactiae, Staphylococcus aureus, Klebsiella pnuemoniae, and Escherichia coli. Overall, butylated hydroxyanisole and tert-butylhydroquinone showed the greatest antimicrobial activity. These phenolics were bactericidal at 250 to 500 ppm against all four bacteria tested. The butylated hydroxytoluene was bactericidal against the gram-positive bacteria but was ineffective against the coliforms. At 250 ppm, disodium ethylenediaminetetraacetic acid was bactericidal against the gram-positive bacteria but much less effective against the gram-negatives. However, diethylene-triaminepentaacetic acid was more growth inhibitorymore » than ethylenediaminetetraacetic acid against the gram-negative bacteria and especially against Escherichia coli. All other compounds were generally much less effective or ineffective against all four microorganisms. Therefore, butylated hydroxyanisole, butylated hydroxytoluene, tert-butylhydroquinone, ethylenediaminetetraacetic acid, and diethylenetriaminepentaacetic acid may have practical implications in the prevention or treatment of bovine mastitis.« less

[Increasing incidence of community-acquired pneumonia caused by atypical microorganisms].

PubMed

Tazón-Varela, M A; Alonso-Valle, H; Muñoz-Cacho, P; Gallo-Terán, J; Piris-García, X; Pérez-Mier, L A

2017-09-01

Knowing the most common microorganisms in our environment can help us to make proper empirical treatment decisions. The aim is to identify those microorganisms causing community-acquired pneumonia. An observational, descriptive and prospective study was conducted, including patients over 14 years with a clinical and radiographic diagnosis of community-acquired pneumonia during a 383 consecutive day period. A record was made of sociodemographic variables, personal history, prognostic severity scales, progress, and pathogenic agents. The aetiological diagnosis was made using blood cultures, detection of Streptococcus pneumoniae and Legionella pneumophila urinary antigens, sputum culture, influenza virus and Streptococcus pyogenes detection. Categorical variables are presented as absolute values and percentages, and continuous variables as their means and standard deviations. Of the 287 patients included in the study (42% women, mean age 66±22 years), 10.45% died and 70% required hospital admission. An aetiological diagnosis was achieved in 43 patients (14.98%), with 16 microorganisms found in 59 positive samples. The most frequently isolated pathogen was Streptococcus pneumonia (24/59, 41%), followed by gram-negative enteric bacilli, Klebsiella pneumonia, Escherichia coli, Serratia marcescens and Enterobacter cloacae isolated in 20% of the samples (12/59), influenza virus (5/59, 9%), methicillin-resistant Staphylococcus aureus (3/59, 5%), Pseudomonas aeruginosa (2/59, 3%), Moraxella catarrhalis (2/59, 3%), Legionella pneumophila (2/59, 3%), and Haemophilus influenza (2/59, 3%). Polymicrobial infections accounted for 14% (8/59). A high percentage of atypical microorganisms causing community-acquired pneumonia were found. Copyright © 2016 Sociedad Española de Médicos de Atención Primaria (SEMERGEN). Publicado por Elsevier España, S.L.U. All rights reserved.

Antibiotic susceptibility of isolates from paediatric intensive care units in Zagreb.

PubMed

Bedenić, Branka; Prahin, Esmina; Vranić-Ladavac, Mirna; Atalić, Vlasta; Sviben, Mario; Frančula-Zaninović, Sonja; Plečko, Vanda; Kalenić, Smilja

2014-02-01

Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) Program is a longitudinal global surveillance study to monitor in vitro data on microbial susceptibility in centers that prescribe meropenem. Results of the six years period (2002-2007) for the antimicrobial efficacy of meropenem compared to other broad-spectrum agents against Gram-negative and Gram-positive isolates collected at pediatric intensive care units of the University Hospital Center Zagreb in Croatia were reported. A total of 110 Gram-negative and 43 Gram-positive pathogens from pediatric specimens were tested. The minimum-inhibitory concentrations (MICs) were determined by broth microdilution method according to CLSI. There was no resistance to either imipenem or meropenem observed for Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis. High resistance rates of K. pneumoniae to ceftazidime and gentamicin (50%) are a raising concern. Pseudomonas aeruginosa was the most resistant Gram-negative species with two (12%) of the strains resistant to meropenem, three (18%) to imipenem, 10 (47%) to gentamicin and six (35%) to piperacillin/tazobactam and ciprofloxacin. According to our results meropenem remains an appropriate antibiotic for the treatment of severe infections caused by Gram-negative bacteria in pediatric population. The results indicate that meropenem has excellent potency and spectrum of activity despite being prescribed for a long time for the treatment of seriously ill patients, and still appears to be a reliable option for the initial empirical therapy of serious nosocomial infections in children. However, later studies have shown the emergence of carbapenem-resistant Gram-negative bacteria after 2008.

Amphibian pathogens in Southeast Asian frog trade.

PubMed

Gilbert, Martin; Bickford, David; Clark, Leanne; Johnson, Arlyne; Joyner, Priscilla H; Ogg Keatts, Lucy; Khammavong, Kongsy; Nguyễn Văn, Long; Newton, Alisa; Seow, Tiffany P W; Roberton, Scott; Silithammavong, Soubanh; Singhalath, Sinpakhone; Yang, Angela; Seimon, Tracie A

2012-12-01

Amphibian trade is known to facilitate the geographic spread of pathogens. Here we assess the health of amphibians traded in Southeast Asia for food or as pets, focusing on Batrachochytrium dendrobatidis (Bd), ranavirus and general clinical condition. Samples were collected from 2,389 individual animals at 51 sites in Lao PDR, Cambodia, Vietnam and Singapore for Bd screening, and 74 animals in Cambodia and Vietnam for ranavirus screening. Bd was found in one frog (n = 347) in Cambodia and 13 in Singapore (n = 419). No Bd was found in Lao PDR (n = 1,126) or Vietnam (n = 497), and no ranavirus was found in Cambodia (n = 70) or Vietnam (n = 4). Mild to severe dermatological lesions were observed in all East Asian bullfrogs Hoplobatrachus rugolosus (n = 497) sampled in farms in Vietnam. Histologic lesions consistent with sepsis were found within the lesions of three frogs and bacterial sepsis in two (n = 4); one had Gram-negative bacilli and one had acid-fast organisms consistent with mycobacterium sp. These results confirm that Bd is currently rare in amphibian trade in Southeast Asia. The presence of Mycobacterium-associated disease in farmed H. rugolosus is a cause for concern, as it may have public health implications and indicates the need for improved biosecurity in amphibian farming and trade.

A randomized, double-blind, placebo-controlled trial of selective digestive decontamination in a medical-surgical intensive care unit.

PubMed

Wiener, J; Itokazu, G; Nathan, C; Kabins, S A; Weinstein, R A

1995-04-01

A randomized, double-blind, placebo-controlled trial of selective decontamination of the oropharynx and gastrointestinal tract was conducted on 61 intubated patients in a medical-surgical intensive care unit (ICU) to determine the impact on nosocomial pneumonia, other infections, and emergence of colonization or infection with antibiotic-resistant bacteria. Over 8 months, 30 patients received an oral paste and solution containing polymyxin, gentamicin, and nystatin; 31 patients received a placebo paste and solution. At study entry, patients in both groups were seriously ill (mean acute physiologic score, 27.2), frequently had pulmonary infiltrates (73.8%), and were likely to be receiving systemic antibiotics (86.9%). There were no differences between study patients and control patients in these characteristics or in frequency of any nosocomial infection (50% vs. 55%), nosocomial pneumonia (27% vs. 26%), febrile days (2.3 vs. 2.0), duration of antibiotic therapy (14.0 vs. 13.4), or mortality rates (37% vs. 48%). There was no difference in infections caused by antibiotic-resistant gram-negative bacilli, although a trend towards more frequent infection with gentamicin-resistant enterococci was found for study patients. Selective decontamination did not appear to be effective in our very ill medical-surgical ICU patients, although the number of patients in our trial was sufficient to detect only a 50% or greater reduction in pneumonia rates.

Anaerobic Bacteria in Clinical Specimens - Frequent, But a Neglected Lot: A Five Year Experience at a Tertiary Care Hospital.

PubMed

Shenoy, Padmaja Ananth; Vishwanath, Shashidhar; Gawda, Ashwini; Shetty, Seema; Anegundi, Renuka; Varma, Muralidhar; Mukhopadhyay, Chiranjay; Chawla, Kiran

2017-07-01

Anaerobic bacteria which constitute a significant proportion of the normal microbiota also cause variety of infections involving various anatomic sites. Considering the tedious culture techniques with longer turnaround time, anaerobic cultures are usually neglected by clinicians and microbiologists. To study the frequency of isolation of different anaerobic bacteria from various clinical specimens. A retrospective study to analyse the frequency of isolation of different anaerobic bacteria, was conducted over a period of five years from 2011 to 2015 including various clinical specimens submitted to anaerobic division of Microbiology laboratory. Anaerobic bacteria were isolated and identified following standard bacteriological techniques. Pathogenic anaerobes (n=336) were isolated from 278 (12.48%) of overall 2227 specimens processed with an average yield of 1.2 isolates. Anaerobes were isolated as polymicrobial flora with or without aerobic bacterial pathogens in 159 (57.2%) patients. Anaerobic Gram-negative bacilli (140, 41.7%) were the predominant isolates. B. fragilis group (67, 19.9%) were the most commonly isolated anaerobic pathogens. Anaerobes were predominantly isolated from deep seated abscess (23.9%). Pathogenic anaerobes were isolated from various infection sites. Unless culture and susceptibility tests are performed as a routine, true magnitude of antimicrobial resistance among anaerobic pathogens will not be known. Knowledge of the distribution of these organisms may assist in the selection of appropriate empirical therapy for anaerobic infections.

Clinico-microbiological study of Pseudomonas aeruginosa in wound infections and the detection of metallo-β-lactamase production.

PubMed

Bangera, Divya; Shenoy, Suchitra M; Saldanha, Dominic Rm

2016-12-01

Pseudomonas aeruginosa is a common opportunistic pathogen of humans among the Gram-negative bacilli. Clinically, it is associated with nosocomial infections like burns and surgical-site wound infections and remains a major health concern, especially among critically ill and immunocompromised patients. This is a prospective laboratory-based 2 year study conducted to isolate P. aeruginosa from wound specimens and the antimicrobial susceptibility pattern with reference to metallo-β-lactamase (MBL) production. Two hundred and twenty-four samples of P. aeruginosa isolated from wound specimens were included in the study. Antimicrobial susceptibility was done as per Clinical Laboratory Standard Institute (CLSI) guidelines. MBL-producing P. aeruginosa was detected using the EDTA disk diffusion synergy test. Statistical analysis was done using the SPSS 11 package (SPSS Inc., Chicago, IL). Out of the 224 P. aeruginosa isolates, 100% were susceptible to polymyxin B and colistin, 92·8% were sensitive to imipenem, 38% showed resistance to gentamicin followed by ceftazidime (31·69%) and meropenem (33·03). Sixteen (7·14%) isolates showed MBL production. Infection caused by drug-resistant P. aeruginosa is important to identify as it poses a therapeutic problem and is also a serious concern for infection control management. The acquired resistance genes can be horizontally transferred to other pathogens or commensals if aseptic procedures are not followed. © 2015 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

Phthalocyanine-sulfonamide conjugates: Synthesis and photodynamic inactivation of Gram-negative and Gram-positive bacteria.

PubMed

da Silva, Raquel Nunes; Cunha, Ângela; Tomé, Augusto C

2018-06-25

Phthalocyanines bearing four or eight sulfonamide units were synthesized and their efficiency in the photodynamic inactivation of Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria was evaluated. Conjugates with simpler sulfonamide units (N,N-diethylbenzenesulfonamide, N-isopropylbenzenesulfonamide and N-(4-methoxyphenyl)benzenesulfonamide) caused stronger inactivation than those with heterocyclic groups (N-(thiazol-2-yl)benzenesulfonamide) or long alkyl chains (N-dodecylbenzenesulfonamide) in both bacteria. Furthermore, the encapsulation of the phthalocyanine-sulfonamide conjugates within polyvinylpyrrolidone micelles, used as drug delivery vehicles, in general showed to enhance the inactivation efficiency. The results show that encapsulated phthalocyanine-sulfonamide conjugates are a promising class of photosensitizers to be used in photodynamic antimicrobial therapy. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

AFRRI reports. First Quarter, January-March 1991. Scientific report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Not Available

1991-04-01

Ionizing radiation reduces the host's defenses to infection (1) and enhances its susceptibility to systemic infection due to endogenous and exogenous organisms. Klebsiella pneumoniae is one of the most frequent causes of gram-negative bacteremia, and is especially prevalent in immunocompromised patients. Therapy for severe systemic infection due to gram-negative bacteria generally involves the use of aminoglycosides in combination with beta lactam antibiotics. However, several recently developed quinolone compounds have exhibited high in vitro bactericidal activity against most gram negative bacteria. including K. pneumoniae. In this study, the authors evaluated the efficacy of oral therapy with several quinolones in a modelmore » of experimental septicemia due to orally ingested K pneumoniae in irradiated mice.« less

Intrinsic, adaptive and acquired antimicrobial resistance in Gram-negative bacteria.

PubMed

Arzanlou, Mohsen; Chai, Wern Chern; Venter, Henrietta

2017-02-28

Gram-negative bacteria are responsible for a large proportion of antimicrobial-resistant infections in humans and animals. Among this class of bacteria are also some of the most successful environmental organisms. Part of this success is their adaptability to a variety of different niches, their intrinsic resistance to antimicrobial drugs and their ability to rapidly acquire resistance mechanisms. These mechanisms of resistance are not exclusive and the interplay of several mechanisms causes high levels of resistance. In this review, we explore the molecular mechanisms underlying resistance in Gram-negative organisms and how these different mechanisms enable them to survive many different stress conditions. © 2017 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Integrating rapid diagnostics and antimicrobial stewardship improves outcomes in patients with antibiotic-resistant Gram-negative bacteremia.

PubMed

Perez, Katherine K; Olsen, Randall J; Musick, William L; Cernoch, Patricia L; Davis, James R; Peterson, Leif E; Musser, James M

2014-09-01

An intervention for Gram-negative bloodstream infections that integrated mass spectrometry technology for rapid diagnosis with antimicrobial stewardship oversight significantly improved patient outcomes and reduced hospital costs. As antibiotic resistance rates continue to grow at an alarming speed, the current study was undertaken to assess the impact of this intervention in a challenging patient population with bloodstream infections caused by antibiotic-resistant Gram-negative bacteria. A total of 153 patients with antibiotic-resistant Gram-negative bacteremia hospitalized prior to the study intervention were compared to 112 patients treated post-implementation. Outcomes assessed included time to optimal antibiotic therapy, time to active treatment when inactive, hospital and intensive care unit length of stay, all-cause 30-day mortality, and total hospital expenditures. Integrating rapid diagnostics with antimicrobial stewardship improved time to optimal antibiotic therapy (80.9 h in the pre-intervention period versus 23.2 h in the intervention period, P < 0.001) and effective antibiotic therapy (89.7 h versus 32 h, P < 0.001). Patients in the pre-intervention period had increased duration of hospitalization compared to those in the intervention period (23.3 days versus 15.3 days, P = 0.0001) and longer intensive care unit length of stay (16 days versus 10.7 days, P = 0.008). Mortality among patients during the intervention period was lower (21% versus 8.9%, P = 0.01) and our study intervention remained a significant predictor of survival (OR, 0.3; 95% confidence interval [CI], 0.12-0.79) after multivariate logistic regression. Mean hospital costs for each inpatient survivor were reduced $26,298 in the intervention cohort resulting in an estimated annual cost savings of $2.4 million (P = 0.002). Integration of rapid identification and susceptibility techniques with antimicrobial stewardship resulted in significant improvements in clinical and financial outcomes for patients with bloodstream infections caused by antibiotic-resistant Gram-negatives. The intervention decreased hospital and intensive care unit length of stay, total hospital costs, and reduced all-cause 30-day mortality. Copyright © 2014. Published by Elsevier Ltd.

Urinary tract infections following radical cystectomy and urinary diversion: a review of 1133 patients.

PubMed

Clifford, Thomas G; Katebian, Behrod; Van Horn, Christine M; Bazargani, Soroush T; Cai, Jie; Miranda, Gus; Daneshmand, Siamak; Djaladat, Hooman

2018-05-01

To investigate the incidence and microbiology of urinary tract infection (UTI) within 90 days following radical cystectomy (RC) and urinary diversion. We reviewed 1133 patients who underwent RC for bladder cancer at our institution between 2003 and 2013; 815 patients (72%) underwent orthotopic diversion, 274 (24%) ileal conduit, and 44 (4%) continent cutaneous diversion. 90-day postoperative UTI incidence, culture results, antibiotic sensitivity/resistance and treatment were recorded through retrospective review. Fisher's exact test, Kruskal-Wallis test, and multivariable analysis were performed. A total of 151 urinary tract infections were recorded in 123 patients (11%) during the first 90 days postoperatively. 21/123 (17%) had multiple infections and 25 (20%) had urosepsis in this time span. Gram-negative rods were the most common etiology (54% of positive cultures). 52% of UTI episodes led to readmission. There was no significant difference in UTI rate, etiologic microbiology (Gram-negative rods, Gram-positive cocci, fungi), or antibiotic sensitivity and resistance patterns between diversion groups. Resistance to quinolones was evident in 87.5% of Gram-positive and 35% of Gram-negative bacteria. In multivariable analysis, Charlson Comorbidity Index > 2 was associated with higher 90-day UTI rate (OR = 1.8, 95% CI 1.1-2.9, p = 0.05) and Candida UTI (OR 5.6, 95% CI 1.6-26.5, p = 0.04). UTI is a common complication and cause of readmission following radical cystectomy and urinary diversion. These infections are commonly caused by Gram-negative rods. High comorbidity index is an independent risk factor for postoperative UTI, but diversion type is not.

Triton Hodge Test: Improved Protocol for Modified Hodge Test for Enhanced Detection of NDM and Other Carbapenemase Producers.

PubMed

Pasteran, Fernando; Gonzalez, Lisandro J; Albornoz, Ezequiel; Bahr, Guillermo; Vila, Alejandro J; Corso, Alejandra

2016-03-01

Accurate detection of carbapenemase-producing Gram-negative bacilli is of utmost importance for the control of nosocomial spread and the initiation of appropriate antimicrobial therapy. The modified Hodge test (MHT), a carbapenem inactivation assay, has shown poor sensitivity in detecting the worldwide spread of New Delhi metallo-β-lactamase (NDM). Recent studies demonstrated that NDM is a lipoprotein anchored to the outer membrane in Gram-negative bacteria, unlike all other known carbapenemases. Here we report that membrane anchoring of β-lactamases precludes detection of carbapenemase activity by the MHT. We also show that this limitation can be overcome by the addition of Triton X-100 during the test, which allows detection of NDM. We propose an improved version of the assay, called the Triton Hodge test (THT), which allows detection of membrane-bound carbapenemases with the addition of this nonionic surfactant. This test was challenged with a panel of 185 clinical isolates (145 carrying known carbapenemase-encoding genes and 40 carbapenemase nonproducers). The THT displayed test sensitivity of >90% against NDM-producing clinical isolates, while improving performance against other carbapenemases. Ertapenem provided the highest sensitivity (97 to 100%, depending on the type of carbapenemase), followed by meropenem (92.5 to 100%). Test specificity was not affected by the addition of Triton (87.5% and 92.5% with ertapenem and meropenem, respectively). This simple inexpensive test confers a large improvement to the sensitivity of the MHT for the detection of NDM and other carbapenemases. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

The First Report of Drug Resistant Bacteria Isolated from the Brown-Banded Cockroach, Supella longipalpa, in Ahvaz, South-western Iran

PubMed Central

Vazirianzadeh, Babak; Dehghani, Rouhullah; Mehdinejad, Manijeh; Sharififard, Mona; Nasirabadi, Nersi

2014-01-01

Background The brown-banded cockroach, Supella longipalpa is known as a carrier of pathogenic bacteria in urban environments, but its role is not well documented regarding the carriage of antibiotic-resistant pathogenic bacteria in Iran. The aim of this study was to determine the resistance bacteria isolated from the brown-banded cockroach in Ahvaz, south west of Iran. Methods: Totally 39 cockroaches were collected from kitchen area of houses and identified. All specimens were cultured to isolate the bacterial agents on blood agar and MacConky agar media. The microorganisms were identified using necessary differential and biochemical tests. Antimicrobial susceptibility tests were performed for isolated organisms by Kirby-Bauer’s disk diffusion according to NCLI guideline, using 18 antibiotics. Results: From the 39 collected S. langipalpa, 179 bacterial agents were isolated, 92 of alimentary ducts and 87 of external body surfaces. Isolated bacteria from cockroaches were identified as Enterobacter spp., Klebsiella spp., Citrobacter spp., Escherichia coli, Salmonella spp., Proteus spp., coagulase negative staphylococci, Serratia marcescens, Staphylococcus aureus, and Bacillus species. The pattern resistance rates were determined for gram negative bacilli and gram positive cocci regarding 18 antibiotics. Conclusion: The brown-banded cockroach can be involved in the spread of drug resistant bacteria and increases the possibility of contacting human environment to drug resistant bacteria. Therefore, the potential of removing this insect should be improved. This is the first original report of drug resistant bacteria isolated from the brown-banded cockroach of Iran. PMID:25629065

Comparative activity of carbapenem testing (the COMPACT study) in Turkey

PubMed Central

2012-01-01

Background Recent evidence indicates that Gram-negative bacterial pathogens, the most common of which are Pseudomonas spp., Enterobacteriaceae, and Acinetobacter baumannii, are frequent causes of hospital-acquired infections. This study aims to evaluate the in vitro activity of doripenem and comparator carbapenem antibiotics against Gram-negative clinical isolates collected from COMParative Activity of Carbapenem Testing (COMPACT) study centres in Turkey. Methods Ten centres in Turkey were invited to submit Pseudomonas aeruginosa, Enterobacteriaceae, and other Gram-negative isolates from intensive care unit (ICU)/non-ICU patients with complicated intra-abdominal infections, bloodstream infections, or nosocomial pneumonia, including ventilator-associated pneumonia, between May and October 2008. Susceptibility was determined by each centre using E-test. A central laboratory performed species confirmation as well as limited susceptibility and quality-control testing. Results Five hundred and ninety six isolates were collected. MIC90 values for doripenem, meropenem, and imipenem, respectively, were 32, ≥ 64, and ≥ 64 mg/L against Pseudomonas spp.; 0.12, 0.12, and 0.5 mg/L against Enterobacteriaceae; and ≥ 64 mg/L for each against other Gram-negative isolates. In determining the susceptibility of hospital isolates of selected Gram-negative pathogens to doripenem, imipenem, and meropenem, we found that against all pathogens combined, the MIC90 for ICU compared with non-ICU isolates was higher. Conclusions Doripenem showed similar or slightly better activity than meropenem and better activity than imipenem against the Gram-negative pathogens collected in Turkey. PMID:22340940

Identification of mycobacterium tuberculosis in sputum smear slide using automatic scanning microscope

NASA Astrophysics Data System (ADS)

Rulaningtyas, Riries; Suksmono, Andriyan B.; Mengko, Tati L. R.; Saptawati, Putri

2015-04-01

Sputum smear observation has an important role in tuberculosis (TB) disease diagnosis, because it needs accurate identification to avoid high errors diagnosis. In development countries, sputum smear slide observation is commonly done with conventional light microscope from Ziehl-Neelsen stained tissue and it doesn't need high cost to maintain the microscope. The clinicians do manual screening process for sputum smear slide which is time consuming and needs highly training to detect the presence of TB bacilli (mycobacterium tuberculosis) accurately, especially for negative slide and slide with less number of TB bacilli. For helping the clinicians, we propose automatic scanning microscope with automatic identification of TB bacilli. The designed system modified the field movement of light microscope with stepper motor which was controlled by microcontroller. Every sputum smear field was captured by camera. After that some image processing techniques were done for the sputum smear images. The color threshold was used for background subtraction with hue canal in HSV color space. Sobel edge detection algorithm was used for TB bacilli image segmentation. We used feature extraction based on shape for bacilli analyzing and then neural network classified TB bacilli or not. The results indicated identification of TB bacilli that we have done worked well and detected TB bacilli accurately in sputum smear slide with normal staining, but not worked well in over staining and less staining tissue slide. However, overall the designed system can help the clinicians in sputum smear observation becomes more easily.

Cross-contamination potential of saliva ejectors used in dentistry.

PubMed

Barbeau, J; ten Bokum, L; Gauthier, C; Prévost, A P

1998-12-01

It has been postulated that evacuation systems used in dentistry could be a source of cross-contamination between patients through backflow of bacteria dislodged from the saliva ejector tubings. The bacterial microflora associated with these systems was characterized using transmission electron microscopy (TEM) and microbiological cultures. The potential for backflow was investigated by a study of pressure differentials in evacuation system tubing and by the presence of bacteria in backflow samples. Evacuation lines were coated with microbial biofilms in which microcolonies of Gram-positive cocci and Gram-negative bacilli predominated, embedded in an extensive polysaccharide matrix. Most bacteria were metabolically active. Occasionally, buccal material such as collagen, fibrin and eukaryotic cell debris was observed. In other experiments, flow reversal was detected several times during saliva ejector use though each of these events was brief (less than 0.1 s). Aspiration of saliva, or occlusion of the mouthpiece opening by the oral mucosa, were the major factors leading to backflow episodes. Bacteria associated with backflow were found in almost 25% assays, with counts ranging from 1-300 cfu/occurrence. The majority of the bacteria isolated from biofilm or backflow samples were staphylococci, micrococci and non-fermentive Gram-negative rods. Pathogens such as Pseudomonas aeruginosa and Staphylococcus aureus were also isolated from backflow fluids. No oral streptococci could be recovered from biofilms in the tubing beyond 15 min from the last saliva ejector use however, suggesting that these species did not survive in the biofilms. These data suggest, although without direct proof of cross-contamination, the possible existence of an infectious risk associated with oral evacuation systems, as potential pathogens may be shed from tubing biofilms following backflow. Even if the risk of cross-contamination between patients is considered to be low, the necessity for regular disinfection of these systems must be stressed, since biofilms can serve as a reservoir for pathogens or harbor potentially infectious material.

Antimicrobial susceptibility pattern in diabetic foot ulcer: a pilot study.

PubMed

Sekhar, Sm; Vyas, N; Unnikrishnan, Mk; Rodrigues, Gs; Mukhopadhyay, C

2014-09-01

Diabetic foot infections (DFIs) are major public health problems and knowledge of microbes that cause infections are helpful to determine proper antibiotic therapy. The aim was to investigate the antimicrobial susceptibility pattern of microbes in DFIs. A cross-sectional study was conducted for a period of 6 months at the Department of General Surgery, KMC hospital, Manipal University, Manipal, India. During this period, 108 patients having DFIs admitted in the general surgery wards were tracked from the hospital data management system. These patients' pus samples were examined as Gram-stained smear and cultured aerobically on blood agar and MacConkey agar plates. Antimicrobial susceptibility test was performed by disc diffusion techniques according to Clinical and Laboratory Standards Institute guidelines. Of the 108 specimens of the diabetic foot lesions, culture showed polymicrobial growth in 44.4% (48/108). Prevalence of Gram-negative organisms (56%, 84/150) was found to be more than Gram-positive organisms (44%, 66/150). However, Staphylococcus aureus was the most frequent pathogen (28%, 42/150). All Gram-positive aerobes were sensitive to doxycycline. All Gram-negative isolates, including extended spectrum beta lactamase producing strains of Proteus mirabilis and Klebsiella oxytoca except Acinetobacter were highly sensitive to amikacin, cefoperazone/sulbactam, and meropenem. Acinetobacter was completely resistant to all the common antibiotics tested. Prevalence showed Gram-negative bacteria was slightly more than Gram-positive bacteria in diabetic foot ulcers. This study recommends doxycycline should be empirical treatment of choice for Gram-positive isolates and amikacin, cefoperazone/sulbactam, and meropenem should be considered for most of the Gram-negatives aerobes.

Diagnosis of nosocomial pneumonia in intubated, intensive care unit patients.

PubMed

Salata, R A; Lederman, M M; Shlaes, D M; Jacobs, M R; Eckstein, E; Tweardy, D; Toossi, Z; Chmielewski, R; Marino, J; King, C H

1987-02-01

The clinical distinction between bacterial colonization of the tracheobronchial tree and nosocomial pneumonia is difficult, especially in intubated patients. We studied 51 intubated, intensive care unit patients prospectively by serial examinations of tracheal aspirates for elastin fibers, graded Gram's stains, and quantitative bacterial cultures in conjunction with clinical and radiologic observations in an attempt to develop criteria for the early detection of pulmonary infection. Patients with infection had new or progressive pulmonary infiltrates plus 1 of the following: positive blood culture results, radiographic evidence of cavitation, or histologic evidence of pneumonia, or 2 or more of the following: new fever, new leukocytosis, or grossly purulent tracheal aspirates. Twenty-one patients developed infection, 22 remained colonized, and 8 had an uncertain status. Infiltrates developed in 34 patients (21 infected, 8 colonized, 5 uncertain status). Gram-negative bacilli were most commonly isolated and were more frequent in infected patients (81 versus 47%, p less than 0.05); Pseudomonas aeruginosa and Serratia marcescens were most often associated with infection. No differences were observed between infected and colonized patients in demographic features, smoking history, underlying disease, previous antibiotic therapy, days in hospital before intubation, preexisting pneumonia upon intubation, or highest temperature or leukocyte count during course. By univariate analysis, infected patients had a longer duration of intubation (p less than 0.05), higher Gram's stain grading for neutrophils (p less than 0.05) or bacteria (p less than 0.005), higher bacterial colony counts (p less than 0.05), and more frequent detection of elastin fibers in tracheal aspirates (p less than 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of antibiotic use on antimicrobial antibiotic resistance and late-onset neonatal infections over 25 years in an Australian tertiary neonatal unit.

PubMed

Carr, David; Barnes, Elizabeth Helen; Gordon, Adrienne; Isaacs, David

2017-05-01

Antibiotic resistance is a worldwide problem. We describe 25 years of responsible antibiotic use in a tertiary neonatal unit. Data on neonatal infections and antibiotic use were collected prospectively from 1990 to 2014 at a single tertiary Sydney neonatal intensive care unit attached to a maternity unit. There are approximately 5500 deliveries and 900 nursery admissions per year. The mean annual rate of late-onset sepsis was 1.64 episodes per 100 admissions. The mean number of late-onset sepsis episodes per admission to the neonatal unit decreased by 4.0% per year (95% CI 2.6% to 5.4%; p<0.0001) and occurred particularly in infants born weighing <1500 g. No infants with negative cultures relapsed with sepsis when antibiotics were stopped after 48-72 hours. Antibiotic use decreased with time. The proportion of colonising methicillin-resistant Staphylococcus aureus isolates decreased by 7.4% per year (95% CI 0.2% to 14.1%; p=0.043). The proportion of colonising Gram-negative bacilli isolates resistant to either third-generation cephalosporins or gentamicin increased by 2.9% per year (95% CI 1.0% to 4.9%; p=0.0035). Most were cephalosporin-resistant; gentamicin resistance was rare. An average of one baby per year died from late-onset sepsis, the rate not varying significantly over time. The mortality from episodes of late-onset sepsis was 25 of 332 (7.5%). Stopping antibiotics after 2-3 days if neonatal systemic cultures are negative is safe. However, it does not prevent the emergence of cephalosporin-resistant Gram-negative organisms. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

A field study evaluation of Petrifilm™ plates as a 24-h rapid diagnostic test for clinical mastitis on a dairy farm.

PubMed

Mansion-de Vries, Elisabeth Maria; Knorr, Nicole; Paduch, Jan-Hendrik; Zinke, Claudia; Hoedemaker, Martina; Krömker, Volker

2014-03-01

Clinical mastitis is one of the most common and expensive diseases of dairy cattle. To make an informed treatment decision, it is important to know the causative pathogen. However, no detection of bacterial growth can be made in approximately 30% of all clinical cases of mastitis. Before selecting the treatment regimen, it is important to know whether the mastitis-causing pathogen (MCP) is Gram-positive or Gram-negative. The aim of this field study was to investigate whether using two 3M Petrifilm™ products on-farm (which conveys a higher degree of sample freshness but also bears a higher risk for contamination than working in a lab) as 24-h rapid diagnostic of clinical mastitis achieved results that were comparable to the conventional microbiological diagnostic method. AerobicCount (AC)-Petrifilm™ and ColiformCount (CC)-Petrifilm™ were used to identify the total bacterial counts and Gram-negative bacteria in samples from clinical mastitis cases, respectively. Missing growth on both plates was classified as no bacterial detection. Growth only on the AC-Petrifilm™ was assessed as Gram-positive, and growth on both Petrifilm™ plates was assessed as Gram-negative bacterial growth. Additionally, milk samples were analysed by conventional microbiological diagnostic method on aesculin blood agar as a reference method. Overall, 616 samples from clinical mastitis cases were analysed. Using the reference method, Gram-positive and Gram-negative bacteria, mixed bacterial growth, contaminated samples and yeast were determined in 32.6%, 20.0%, 2.5%, 14.1% and 1.1% of the samples, respectively. In 29.7% of the samples, microbiological growth could not be identified. Using the Petrifilm™ concept, bacterial growth was detected in 59% of the culture-negative samples. The sensitivity of the Petrifilm™ for Gram-positive and Gram-negative MCP was 85.2% and 89.9%, respectively. The specificity was 75.4% for Gram-positive and 88.4% for Gram-negative MCP. For the culture-negative samples, sensitivity was 41.0% and specificity was 91.0%. The results indicate that the Petrifilm™ concept is suitable for therapeutic decision-making at the farm level or in veterinary practice. As this concept does not allow any statement about the genus or species of microorganisms, relevant MCP should be assessed periodically at the herd level with conventional microbiological diagnostics. Copyright © 2014 Elsevier B.V. All rights reserved.

Gram-negative bacteraemia; a multi-centre prospective evaluation of empiric antibiotic therapy and outcome in English acute hospitals.

PubMed

Fitzpatrick, J M; Biswas, J S; Edgeworth, J D; Islam, J; Jenkins, N; Judge, R; Lavery, A J; Melzer, M; Morris-Jones, S; Nsutebu, E F; Peters, J; Pillay, D G; Pink, F; Price, J R; Scarborough, M; Thwaites, G E; Tilley, R; Walker, A S; Llewelyn, M J

2016-03-01

Increasing antibiotic resistance makes choosing antibiotics for suspected Gram-negative infection challenging. This study set out to identify key determinants of mortality among patients with Gram-negative bacteraemia, focusing particularly on the importance of appropriate empiric antibiotic treatment. We conducted a prospective observational study of 679 unselected adults with Gram-negative bacteraemia at ten acute english hospitals between October 2013 and March 2014. Appropriate empiric antibiotic treatment was defined as intravenous treatment on the day of blood culture collection with an antibiotic to which the cultured organism was sensitive in vitro. Mortality analyses were adjusted for patient demographics, co-morbidities and illness severity. The majority of bacteraemias were community-onset (70%); most were caused by Escherichia coli (65%), Klebsiella spp. (15%) or Pseudomonas spp. (7%). Main foci of infection were urinary tract (51%), abdomen/biliary tract (20%) and lower respiratory tract (14%). The main antibiotics used were co-amoxiclav (32%) and piperacillin-tazobactam (30%) with 34% receiving combination therapy (predominantly aminoglycosides). Empiric treatment was inappropriate in 34%. All-cause mortality was 8% at 7 days and 15% at 30 days. Independent predictors of mortality (p <0.05) included older age, greater burden of co-morbid disease, severity of illness at presentation and inflammatory response. Inappropriate empiric antibiotic therapy was not associated with mortality at either time-point (adjusted OR 0.82; 95% CI 0.35-1.94 and adjusted OR 0.92; 95% CI 0.50-1.66, respectively). Although our study does not exclude an impact of empiric antibiotic choice on survival in Gram-negative bacteraemia, outcome is determined primarily by patient and disease factors. Copyright © 2015 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Aerobiological Stabilities of Different Species of Gram-Negative Bacteria, Including Well-Known Biothreat Simulants, in Single-Cell Particles and Cell Clusters of Different Compositions

PubMed Central

Skogan, Gunnar

2017-01-01

ABSTRACT The ability to perform controlled experiments with bioaerosols is a fundamental enabler of many bioaerosol research disciplines. A practical alternative to using hazardous biothreat agents, e.g., for detection equipment development and testing, involves using appropriate model organisms (simulants). Several species of Gram-negative bacteria have been used or proposed as biothreat simulants. However, the appropriateness of different bacterial genera, species, and strains as simulants is still debated. Here, we report aerobiological stability characteristics of four species of Gram-negative bacteria (Pantoea agglomerans, Serratia marcescens, Escherichia coli, and Xanthomonas arboricola) in single-cell particles and cell clusters produced using four spray liquids (H2O, phosphate-buffered saline[PBS], spent culture medium[SCM], and a SCM-PBS mixture). E. coli showed higher stability in cell clusters from all spray liquids than the other species, but it showed similar or lower stability in single-cell particles. The overall stability was higher in cell clusters than in single-cell particles. The highest overall stability was observed for bioaerosols produced using SCM-containing spray liquids. A key finding was the observation that stability differences caused by particle size or compositional changes frequently followed species-specific patterns. The results highlight how even moderate changes to one experimental parameter, e.g., bacterial species, spray liquid, or particle size, can strongly affect the aerobiological stability of Gram-negative bacteria. Taken together, the results highlight the importance of careful and informed selection of Gram-negative bacterial biothreat simulants and also the accompanying particle size and composition. The outcome of this work contributes to improved selection of simulants, spray liquids, and particle size for use in bioaerosol research. IMPORTANCE The outcome of this work contributes to improved selection of simulants, spray liquids, and particle size for use in bioaerosol research. Taken together, the results highlight the importance of careful and informed selection of Gram-negative bacterial biothreat simulants and also the accompanying particle size and composition. The results highlight how even moderate changes to one experimental parameter, e.g., bacterial species, spray liquid, or particle size, can strongly affect the aerobiological stability of Gram-negative bacteria. A key finding was the observation that stability differences caused by particle size or compositional changes frequently followed species-specific patterns. PMID:28687646

Performance of Gram staining on blood cultures flagged negative by an automated blood culture system.

PubMed

Peretz, A; Isakovich, N; Pastukh, N; Koifman, A; Glyatman, T; Brodsky, D

2015-08-01

Blood is one of the most important specimens sent to a microbiology laboratory for culture. Most blood cultures are incubated for 5-7 days, except in cases where there is a suspicion of infection caused by microorganisms that proliferate slowly, or infections expressed by a small number of bacteria in the bloodstream. Therefore, at the end of incubation, misidentification of positive cultures and false-negative results are a real possibility. The aim of this work was to perform a confirmation by Gram staining of the lack of any microorganisms in blood cultures that were identified as negative by the BACTEC™ FX system at the end of incubation. All bottles defined as negative by the BACTEC FX system were Gram-stained using an automatic device and inoculated on solid growth media. In our work, 15 cultures that were defined as negative by the BACTEC FX system at the end of the incubation were found to contain microorganisms when Gram-stained. The main characteristic of most bacteria and fungi growing in the culture bottles that were defined as negative was slow growth. This finding raises a problematic issue concerning the need to perform Gram staining of all blood cultures, which could overload the routine laboratory work, especially laboratories serving large medical centers and receiving a large number of blood cultures.

Redundancy and modularity in membrane-associated dissimilatory nitrate reduction in Bacillus

PubMed Central

Heylen, Kim; Keltjens, Jan

2012-01-01

The genomes of two phenotypically denitrifying type strains of the genus Bacillus were sequenced and the pathways for dissimilatory nitrate reduction were reconstructed. Results suggest that denitrification proceeds in the periplasmic space and in an analogous fashion as in Gram-negative organisms, yet with the participation of proteins that tend to be membrane-bound or membrane-associated. A considerable degree of functional redundancy was observed with marked differences between B. azotoformans LMG 9581T and B. bataviensis LMG 21833T. In addition to the already characterized menaquinol/cyt c-dependent nitric oxide reductase (Suharti et al., 2001, 2004) of which the encoding genes could be identified now, evidence for another novel nitric oxide reductase (NOR) was found. Also, our analyses confirm earlier findings on branched electron transfer with both menaquinol and cytochrome c as reductants. Quite unexpectedly, both bacilli have the disposal of two parallel pathways for nitrite reduction enabling a life style as a denitrifier and as an ammonifying bacterium. PMID:23087684

In Vitro Studies of Semisynthetic α- (Substituted-Ureido) Penicillins

PubMed Central

Bodey, Gerald P.; Stewart, Dorothy

1971-01-01

The activity of three α-(substituted-ureido) penicillins was evaluated in vitro against 599 clinical isolates of gram-negative bacilli, by use of the broth-dilution technique. At a concentration of 12.5 μg or less/ml, BL-P1597 inhibited 90% of isolates of Pseudomonas sp., 56% of Enterobacter sp., 67% of indole-positive Proteus spp., 72% of Escherichia coli, and 85% of Proteus mirabilis. BL-P1654 had similar activity, whereas BL-P1532 was much less active. At a concentration of 25 μg or less/ml, BL-P1597 also inhibited nearly 60% of isolates of Klebsiella sp. and nearly 40% of Serratia sp. BL-P1597 and BL-P1654 were as active as ampicillin and carbenicillin against E. coli and P. mirabilis. They were less active than carbenicillin against indole-positive Proteus spp. Both drugs were substantially more active than carbenicillin against Pseudomonas sp. A strain of Pseudomonas sp. which developed resistance to carbenicillin also developed resistance to the α-(substituted-ureido) penicillins simultaneously. PMID:4930281

Aerobic vaginal pathogens and their sensitivity pattern.

PubMed

Mumtaz, Shamim; Ahmad, Mumtaz; Aftab, Irum; Akhtar, Naeem; ul Hassan, Masood; Hamid, Abdul

2008-01-01

The vaginal flora is a complicated environment, containing dozens of microbiological species in variable quantities and relative proportions. The frequent cause of vaginal discharge is an infection or colonization with different microorganisms. Some pathologic conditions causing vaginitis are well defined yet, 7-72% of women with vaginitis may remain undiagnosed and such forms of abnormal vaginal flora neither considered as normal, nor can be called bacterial vaginosis have been termed as 'intermediate flora' and its management probably differ from that of bacterial vaginosis. It is of crucial importance in pregnant females at risk of preterm delivery. The present study has been conducted especially to elucidate this type of aerobic vaginal isolates and their culture and sensitivity towards currently used antibiotics. This study was conducted at the Microbiology Department of Fauji Foundation Hospital, Rawalpindi over a period of two years (April 2004-March 2006). One thousand, nine hundred and twenty three high vaginal swabs, both from indoor and outdoor patients were collected, cultured and their susceptibility to various antibiotics was determined. Significant growth was obtained in 731 samples. The highest frequency of infection (39.5%) was observed at 31-40 years followed by 41-50 years (35.8%). About 76% were from outdoor and 24% were from indoor patients. Staphylococcus aureus was the most prevalent vaginal pathogen at 11-60 yrs & with highest prevalence at 31-40 years followed by 41-50 years. It was a predominant pathogen in both indoor (35%) as well as outdoor (41.6%) patients, followed by enteric gram-negative bacilli and other gram-positive cocci. There were very few antibiotics among the conventionally available aminoglycosides, third generation cephalosporins, penicillin, quinolones, sulfonamides and tetracyclines possessing good sensitivity (> 80%) against any one the common aerobic vaginal pathogens. The effective chemotherapeutics agents belong to the groups of carbapenems and beta-lactams beta-lactamase inhibitor combinations. The high prevalence of gynaecological infections demands that the patients with gynaecological symptoms be investigated thoroughly. Culture must invariably be done. Currently the antibiotics showing good sensitivity are very expensive. So there is a need for an effective antimicrobial policy. Effective drug should be reserved for the treatment of serious life threatening situations only.

[Estimation of activity of pharmakopeal disinfectants and antiseptics against Gram-negative and Gram-positive bacteria isolated from clinical specimens, drugs and environment].

PubMed

Grzybowska, Wanda; Młynarczyk, Grazyna; Młynarczyk, Andrzej; Bocian, Ewa; Luczak, Mirosław; Tyski, Stefan

2007-01-01

The MIC of nine different disinfectants and antiseptics were determined for the Gram-negative and Gram-positive bacteria. Strains originated from clinical specimens, drugs and environment. A sensitivity was determined against chlorhexidinum digluconate (Gram-negative: 0,625-80 mg/L, Gram-positive: 0,3-10 mg/L), benzalconium chloride (Gram-negative: 2,5-1280 mg/L, Gram-positive: 1,25-20 mg/L), salicilic acid (Gram-negative and Gram-positive: 400-1600 mg/L), benzoic acid (Gram-negative: 800-1600 mg/L, Gram-positive: 400-1 600 mg/L), boric acid (Gram-negative: 800-12 800 mg/L, Gram-positive: 1 600-6400 mg/L), chloramine B (Gram-negative: 1600-6400 mg/L, Gram-positive:800- 6400 mg/L), jodine (Gram-negative: 200-1600 mg/L, Gram-positive: 200-1600 mg/L), etacridine lactate (Gram-negative: 40 do > 20480 mg/L, Gram-positive: 40-1280 mg/L) and resorcine (Gram-negative: 1600-6400 mg/L, Gram-positive: 800-6400 mg/L). Diversified values of MIC for different strains were obtained, especially in the case of benzalconium chloride, etacridine lactate, chlorhexidinum digluconate, boric acid and iodine. Strains isolated from environment were usually more susceptible to examined compounds than clinical strains. The biggest diversification of sensitivity was observed among strains originated from drugs where besides sensitive appeared strains characterizing by very high MIC values of some substances, eg. boric acid.

Comparison of topical mupirocin and gentamicin in the prevention of peritoneal dialysis-related infections: A systematic review and meta-analysis.

PubMed

Tsai, Chia-Chi; Yang, Po-Sheng; Liu, Chien-Liang; Wu, Chih-Jen; Hsu, Yi-Chiung; Cheng, Shih-Ping

2018-01-01

Topical antibiotics have been shown to reduce exit-site infection and peritonitis. The aim of this study was to compare infection rates between mupirocin and gentamicin. Multiple comprehensive databases were searched systematically to include relevant randomized controlled trials and observational studies. Pooled risk ratios (RRs) and 95% confidence intervals were calculated for the incidences of exit-site infection and peritonitis. Seven studies (mupirocin group n = 458, gentamicin group n = 448) were analyzed for exit-site infection. The risk of gram-positive exit-site infection was similar between the groups. Gram-negative exit-site infection rate was higher in the mupirocin group (RR = 2.125, P = 0.037). Six studies were assessed the peritonitis risk. There was no difference in the gram-positive and -negative peritonitis rate. Topical use of gentamicin is associated with fewer exit-site infections caused by gram-negative organisms. Gentamicin has comparable efficacy to mupirocin for peritonitis and gram-positive exit-site infection. Copyright © 2017 Elsevier Inc. All rights reserved.

The clinical efficacy of a clarithromycin-based regimen for Mycobacterium avium complex disease: A nationwide post-marketing study.

PubMed

Kadota, Jun-Ichi; Kurashima, Atsuyuki; Suzuki, Katsuhiro

2017-05-01

The revised 2007 American Thoracic Society/Infectious Diseases Society of America statement recommend clarithromycin-based combination therapy for treatment of Mycobacterium avium complex lung disease and stipulates approximately 1 year of continuous treatment after bacilli negative conversion. However, supporting data are insufficient. Our objective was to obtain data on the clinical outcome of clarithromycin-based daily regimens by conducting a nationwide retrospective post-marketing study of M. avium complex lung disease. In accordance with the Japanese guidelines, patients were enrolled in this survey according to their chest radiographic findings and microbiologic test results. They were treated with a multidrug regimen including clarithromycin, rifampicin, and ethambutol (clarithromycin-based regimen) until bacilli negative conversion, and the treatment was continued for approximately 1 year after the initial conversion. Data were collected before administration, at the time of bacilli negative conversion, at the end of treatment, and at 6 months after the end of treatment. Of the 466 subjects enrolled in the study, 271 patients who received clarithromycin at 800 mg/day underwent evaluation for M. avium complex disease. The final bacilli negative conversion rate in those patients was 94.7%. The bacteriological relapse rate was 5.0% (5/100 patients). Bacteriological relapse was noted in patients treated for less than 15 months after conversion. No life-threatening or serious adverse drug reactions were observed. This study demonstrated that a clarithromycin-based daily regimen can yield a high bacteriological conversion rate in M. avium complex disease. After conversion, treatment for less than 15 months might be insufficient to prevent bacteriological relapse. Copyright © 2017 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Catechol-Functional Chitosan/Silver Nanoparticle Composite as a Highly Effective Antibacterial Agent with Species-Specific Mechanisms.

PubMed

Huang, Xiaofei; Bao, Xiaojiong; Liu, Yalan; Wang, Zhengke; Hu, Qiaoling

2017-05-12

In this study, silver nanoparticles (Ag NPs) coated with catechol-conjugated chitosan (CSS) were prepared using green methods. Interestingly, we uncovered that CSS-coated Ag NPs (CSS-Ag NPs) exhibited a higher toxicity against gram-negative Escherichia coli (E. coli) bacteria than against gram-positive Staphylococcus aureus (S. aureus) bacteria. The differences revealed that the CSS-Ag NPs killed gram bacteria with distinct, species-specific mechanisms. The aim of this study is to further investigate these underlying mechanisms through a series of analyses. The ultrastructure and morphology of the bacteria before and after treatment with CSS-Ag NPs were observed. The results demonstrated the CSS-Ag NPs killed gram-positive bacteria through a disorganization of the cell wall and leakage of cytoplasmic content. In contrast, the primary mechanism of action on gram-negative bacteria was a change in membrane permeability, induced by adsorption of CSS-Ag NPs. The species-specific mechanisms are caused by structural differences in the cell walls of gram bacteria. Gram-positive bacteria are protected from CSS-Ag NPs by a thicker cell wall, while gram-negatives are more easily killed due to an interaction between a special outer membrane and the nanoparticles. Our study offers an in-depth understanding of the antibacterial behaviors of CSS-Ag NPs and provides insights into ultimately optimizing the design of Ag NPs for treatment of bacterial infections.

Corynebacterium species isolated from patients with mastitis.

PubMed

Paviour, Sue; Musaad, Sahar; Roberts, Sally; Taylor, Graeme; Taylor, Susan; Shore, Keith; Lang, Selwyn; Holland, David

2002-12-01

Corynebacteria were isolated from breast tissue, pus, or deep wound swabs of 24 women; the most common species isolated was the newly described Corynebacterium kroppenstedtii, followed by Corynebacterium amycolatum and Corynebacterium tuberculostearicum. Gram-positive bacilli were seen in samples sent for culture or in histological specimens for 12 women, and 9 of the 12 women from whom adequate histological specimens were obtained had conditions that met the criteria for granulomatous lobular mastitis, a chronic inflammatory disease of unknown etiology.

The cost and management of different types of clinical mastitis in dairy cows estimated by dynamic programming.

PubMed

Cha, E; Bar, D; Hertl, J A; Tauer, L W; Bennett, G; González, R N; Schukken, Y H; Welcome, F L; Gröhn, Y T

2011-09-01

The objective of this study was to estimate the cost of 3 different types of clinical mastitis (CM) (caused by gram-positive bacteria, gram-negative bacteria, and other organisms) at the individual cow level and thereby identify the economically optimal management decision for each type of mastitis. We made modifications to an existing dynamic optimization and simulation model, studying the effects of various factors (incidence of CM, milk loss, pregnancy rate, and treatment cost) on the cost of different types of CM. The average costs per case (US$) of gram-positive, gram-negative, and other CM were $133.73, $211.03, and $95.31, respectively. This model provided a more informed decision-making process in CM management for optimal economic profitability and determined that 93.1% of gram-positive CM cases, 93.1% of gram-negative CM cases, and 94.6% of other CM cases should be treated. The main contributor to the total cost per case was treatment cost for gram-positive CM (51.5% of the total cost per case), milk loss for gram-negative CM (72.4%), and treatment cost for other CM (49.2%). The model affords versatility as it allows for parameters such as production costs, economic values, and disease frequencies to be altered. Therefore, cost estimates are the direct outcome of the farm-specific parameters entered into the model. Thus, this model can provide farmers economically optimal guidelines specific to their individual cows suffering from different types of CM. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Discovery of Novel Pyridone-Conjugated Monosulfactams as Potent and Broad-Spectrum Antibiotics for Multidrug-Resistant Gram-Negative Infections.

PubMed

Tan, Liang; Tao, Yunliang; Wang, Ting; Zou, Feng; Zhang, Shuhua; Kou, Qunhuan; Niu, Ao; Chen, Qian; Chu, Wenjing; Chen, Xiaoyan; Wang, Haidong; Yang, Yushe

2017-04-13

Conjugating a siderophore to an antibiotic is a promising strategy to overcome the permeability-mediated resistance of Gram-negative pathogens. On the basis of the structure of BAL30072, novel pyridone-conjugated monosulfactams incorporating diverse substituents into the methylene linker between the 1,3-dihydroxypyridin-4(1H)-one and the aminothiazole oxime were designed and synthesized. Structure-activity relationship studies revealed that a variety of substituents were tolerated, with isopropyl (compound 12c) and methylthiomethyl (compound 16a) showing the best efficacy against multidrug-resistant (MDR) Gram-negative pathogens. In addition, compound 12c exhibits a good free fraction rate in an in vitro human plasma protein binding test, along with a low clearance and favorable plasma exposure in vivo. In a murine systemic infection model with MDR Klebsiella pneumoniae, compound 12c shows an ED 50 of 10.20 mg/kg. Taken together, the results indicate that compound 12c is a promising drug candidate for the treatment of serious infections caused by MDR Gram-negative pathogens.

Bio-fabricated silver nanoparticles preferentially targets Gram positive depending on cell surface charge.

PubMed

Mandal, Debasis; Kumar Dash, Sandeep; Das, Balaram; Chattopadhyay, Sourav; Ghosh, Totan; Das, Debasis; Roy, Somenath

2016-10-01

Recently bio-inspired experimental processes for synthesis of nanoparticles are receiving significant attention in nanobiotechnology. Silver nanoparticles (Ag NPs) have been used very frequently in recent times to the wounds, burns and bacterial infections caused by drug-resistant microorganisms. Though, the antibacterial effects of Ag NPs on some multi drug-resistant bacteria specially against Gram positive bacteria has been established, but further investigation is needed to elicit its effectiveness against Gram negatives and to identify the probable mechanism of action. Thus, the present study was conducted to synthesize Ag NPs using Andrographis paniculata leaf extract and to investigate its antibacterial efficacy. After synthesis process the biosynthesized nanoparticles were purified and characterized with the help of various physical measurement techniques which raveled their purity, stability and small size range. The antimicrobial activity of Ag NPs was determined against both Gram-positive Enterococcus faecalis and Gram-negative Proteus vulgaris. Results showed comparatively higher antibacterial efficacy of Ag NPs against Gram positive Enterococcus faecalis strains. It was found that greater difference in zeta potential values between Gram positive bacteria and Ag NPs triggers better internalization of the particles. Thus the cell surface charge played vital role in cell killing which was confirmed by surface zeta potential study. Finally it may be concluded that green synthesized Ag NPs using Andrographis paniculata leaf extract can be very useful against both multi drug resistant Gram-positive and Gram-negative bacteria. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Effect of extremely low frequency electromagnetic fields on bacterial membrane.

PubMed

Oncul, Sule; Cuce, Esra M; Aksu, Burak; Inhan Garip, Ayse

2016-01-01

The effect of extremely low frequency electromagnetic fields (ELF-EMF) on bacteria has attracted attention due to its potential for beneficial uses. This research aimed to determine the effect of ELF-EMF on bacterial membrane namely the membrane potential, surface potential, hydrophobicity, respiratory activity and growth. Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli were subjected to ELF-EMF, 50 Hz, 1 mT for 2 h. Membrane potential was determined by fluorescence spectroscopy with or without EDTA (Ethylenediaminetetraacetic acid) with DisC3(5) (3,3-dipropylthiacarbocyanine iodide), zeta potential measurements were performed by electrophoretic mobility, hydrophobicity of the membrane was measured with MATH (Microbial Adhesion to Hydrocarbons) test, respiratory activity was determined with CTC (5-Cyano-2,3-ditolyl tetrazolium chloride), colony forming unit (CFU) and DAPI (4',6-diamidino-2-phenylindole, dihydrochloride) was used for growth determinations. ELF-EMF caused changes in physicochemical properties of both Gram-positive and Gram-negative bacteria. Hyperpolarization was seen in S. aureus and EDTA-treated E. coli. Surface potential showed a positive shift in S. aureus contrariwise to the negative shift seen in EDTA-untreated E. coli. Respiratory activity increased in both bacteria. A slight decrease in growth was observed. These results show that ELF-EMF affects the crucial physicochemical processes in both Gram-positive and Gram-negative bacteria which need further research.

Value of PCR in sonication fluid for the diagnosis of orthopedic hardware-associated infections: Has the molecular era arrived?

PubMed

Renz, Nora; Cabric, Sabrina; Morgenstern, Christian; Schuetz, Michael A; Trampuz, Andrej

2018-04-01

Bone healing disturbance following fracture fixation represents a continuing challenge. We evaluated a novel fully automated polymerase chain reaction (PCR) assay using sonication fluid from retrieved orthopedic hardware to diagnose infection. In this prospective diagnostic cohort study, explanted orthopedic hardware materials from consecutive patients were investigated by sonication and the resulting sonication fluid was analyzed by culture (standard procedure) and multiplex PCR (investigational procedure). Hardware-associated infection was defined as visible purulence, presence of a sinus tract, implant on view, inflammation in peri-implant tissue or positive culture. McNemar's chi-squared test was used to compare the performance of diagnostic tests. For the clinical performance all pathogens were considered, whereas for analytical performance only microorganisms were considered for which primers are included in the PCR assay. Among 51 patients, hardware-associated infection was diagnosed in 38 cases (75%) and non-infectious causes in 13 patients (25%). The sensitivity for diagnosing infection was 66% for peri-implant tissue culture, 84% for sonication fluid culture, 71% (clinical performance) and 77% (analytical performance) for sonication fluid PCR, the specificity of all tests was >90%. The analytical sensitivity of PCR was higher for gram-negative bacilli (100%), coagulase-negative staphylococci (89%) and Staphylococcus aureus (75%) than for Cutibacterium (formerly Propionibacterium) acnes (57%), enterococci (50%) and Candida spp. (25%). The performance of sonication fluid PCR for diagnosis of orthopedic hardware-associated infection was comparable to culture tests. The additional advantage of PCR was short processing time (<5 h) and fully automated procedure. With further improvement of the performance, PCR has the potential to complement conventional cultures. Copyright © 2018 Elsevier Ltd. All rights reserved.

Molecular analysis of bacterial communities in raw cow milk and the impact of refrigeration on its structure and dynamics.

PubMed

Raats, Dina; Offek, Maya; Minz, Dror; Halpern, Malka

2011-05-01

The impact of refrigeration on raw cow milk bacterial communities in three farm bulk tanks and three dairy plant silo tanks was studied using two methods: DGGE and cloning. Both methods demonstrated that bacterial taxonomic diversity decreased during refrigeration. Gammaproteobacteria, especially Pseudomonadales, dominated the milk after refrigeration. Farm samples and dairy plant samples differed in their microbial community composition, the former showing prevalence of Gram-positive bacteria affiliated with the classes Bacilli, Clostridia and Actinobacteria, the latter showing prevalence of Gram-negative species belonging to the Gammaproteobacteria class. Actinobacteria prevalence in the farm milk samples immediately after collection stood at about 25% of the clones. A previous study had found that psychrotolerant Actinobacteria identified in raw cow milk demonstrated both lipolytic and proteolytic enzymatic activity. Thus, we conclude that although Pseudomonadales play an important role in milk spoilage after long periods of cold incubation, Actinobacteria occurrence may play an important role when assessing the quality of milk arriving at the dairy plant from different farms. As new cooling technologies reduce the initial bacterial counts of milk to very low levels, more sensitive and efficient methods to evaluate the bacterial quality of raw milk are required. The present findings are an important step towards achieving this goal. Copyright © 2010 Elsevier Ltd. All rights reserved.

Gram-negative, but not Gram-positive, bacteria elicit strong PGE2 production in human monocytes.

PubMed

Hessle, Christina C; Andersson, Bengt; Wold, Agnes E

2003-12-01

Gram-positive and Gram-negative bacteria induce different cytokine patterns in human mononuclear cells. We have seen that Gram-positives preferentially induce IL-12 and TNF-alpha, whereas Gram-negatives induce more IL-10, IL-6, and IL-8. In this study, we compared the capacity of these two groups of bacteria to induce PGE2. Monocytes stimulated with Gram-negative bacterial species induced much more PGE2 than did Gram-positive bacteria (5600 +/- 330 vs. 1700 +/- 670 pg/mL, p < 0.001). Blocking of COX-2 by NS398 abolished PGE2 production, but did not alter the cytokine patterns induced by Gram-positive and Gram-negative bacteria. We suggest that Gram-positive and Gram-negative bacteria may stimulate different innate effector functions; Gram-positive bacteria promoting cell-mediated effector functions whereas Gram-negative bacteria inducing mediators inhibiting the same.

Micro-Raman spectroscopy for identification and classification of UTI bacteria

NASA Astrophysics Data System (ADS)

Yogesha, M.; Chawla, Kiran; Acharya, Mahendra; Chidangil, Santhosh; Bankapur, Aseefhali

2017-07-01

Urinary tract infection (UTI) is one of the major clinical problems known to mankind, especially among adult women. Conventional methods for identification of UTI causing bacteria are time consuming and expensive. Therefore, a rapid and cost-effective method is desired. In the present study, five bacteria (one Gram-positive and four Gram-negative), most commonly known to cause UTI, have been identified and classified using micro-Raman spectroscopy combined with principal component analysis (PCA).

Blackleg in cattle: A case report of fetal infection and a literature review.

PubMed

Abreu, Camila C; Edwards, Erin E; Edwards, John F; Gibbons, Philippa M; Leal de Araújo, Jeann; Rech, Raquel R; Uzal, Francisco A

2017-09-01

Clostridium chauvoei causes blackleg in cattle. The disease has been reported worldwide, and although it can be prevented by vaccination, sporadic cases and occasional outbreaks still occur. We describe a case of blackleg in a 2-y-old, pregnant Gyr cow with in utero transmission to the fetus. The cow had characteristic gross and microscopic lesions of blackleg including widespread necrohemorrhagic and emphysematous skeletal and myocardial myositis, and fibrinous pericarditis. Her uterus contained a near-term, markedly emphysematous fetus with skeletal muscle and myocardial lesions similar to those seen in the dam. Histopathology of dam and fetal tissues revealed numerous gram-positive bacilli, many of them with sub-terminal spores, in multiple tissues. These bacilli were identified as C. chauvoei by immunohistochemistry. Anaerobic culture and fluorescent antibody tests performed on skeletal muscle from both the dam and fetus were positive for C. chauvoei, confirming a diagnosis of blackleg. Blackleg is a so-called endogenous infection, and the currently accepted pathogenesis involves ingestion of spores that are transported to muscle tissues where they lie dormant until anaerobiosis prompts germination. Germinating bacteria are histotoxic, producing severe, local necrosis and ultimately lethal toxemia. This model, however, has not been confirmed experimentally and also fails to explain some cases of the disease. A presumptive diagnosis of blackleg is based on clinical, gross, and histologic findings. Diagnostic confirmation necessitates the detection of C. chauvoei by culture, PCR, or immunodetection methods.

Characterization of fungal antagonistic bacilli isolated from aerial roots of banyan (Ficus benghalensis) using intact-cell MALDI-TOF mass spectrometry (ICMS).

PubMed

Pathak, K V; Keharia, H

2013-05-01

To characterize fungal antagonistic bacilli isolated from aerial roots of banyan tree and identify the metabolites responsible for their antifungal activity. Seven gram positive, endospore-forming, rod-shaped endophytic bacterial strains exhibiting a broad-spectrum antifungal activity were isolated from the surface-sterilized aerial roots of banyan tree. The isolates designated as K1, A2, A4 and A12 were identified as Bacillus subtilis, whereas isolates A11 and A13 were identified as Bacillus amyloliquefaciens using Biolog Microbial Identification System. The antifungal lipopeptides, surfactins, iturins and fengycins with masses varying in the range from m/z 900 to m/z 1550 could be detected using intact-cell MALDI-TOF mass spectrometry (ICMS). On the basis of mass spectral and carbon source utilization profile, all seven endophytes could be distinguished from each other. Furthermore, ICMS analysis revealed higher extent of heterogeneity among iturins and fengycins produced by B. subtilis K1, correlating well with its higher antifungal activity in comparison with other isolates. Seven fungal antagonistic bacilli were isolated from aerial roots of banyan tree, exhibiting broad spectrum of antifungal activity, among which B. subtilis K1 isolate was found to be most potent. The ICMS analysis revealed that all these isolates produced cyclic lipopeptides belonging to surfactin, iturin and fengycin families and exhibited varying degree of heterogeneity. The endophytes are considered as a potential source of novel bioactive metabolites, and this study describes the potent fungal antagonistic bacilli from aerial roots of banyan tree. The isolates described in this study have a prospective application as biocontrol agents. Also ICMS analysis described in this study for characterization of antifungal metabolites produced by banyan endophytic bacilli may be used as a high throughput tool for screening of microbes producing novel cyclic lipopeptides. © 2013 The Society for Applied Microbiology.

Genomic determinants of sporulation in Bacilli and Clostridia: towards the minimal set of sporulation-specific genes.

PubMed

Galperin, Michael Y; Mekhedov, Sergei L; Puigbo, Pere; Smirnov, Sergey; Wolf, Yuri I; Rigden, Daniel J

2012-11-01

Three classes of low-G+C Gram-positive bacteria (Firmicutes), Bacilli, Clostridia and Negativicutes, include numerous members that are capable of producing heat-resistant endospores. Spore-forming firmicutes include many environmentally important organisms, such as insect pathogens and cellulose-degrading industrial strains, as well as human pathogens responsible for such diseases as anthrax, botulism, gas gangrene and tetanus. In the best-studied model organism Bacillus subtilis, sporulation involves over 500 genes, many of which are conserved among other bacilli and clostridia. This work aimed to define the genomic requirements for sporulation through an analysis of the presence of sporulation genes in various firmicutes, including those with smaller genomes than B. subtilis. Cultivable spore-formers were found to have genomes larger than 2300 kb and encompass over 2150 protein-coding genes of which 60 are orthologues of genes that are apparently essential for sporulation in B. subtilis. Clostridial spore-formers lack, among others, spoIIB, sda, spoVID and safA genes and have non-orthologous displacements of spoIIQ and spoIVFA, suggesting substantial differences between bacilli and clostridia in the engulfment and spore coat formation steps. Many B. subtilis sporulation genes, particularly those encoding small acid-soluble spore proteins and spore coat proteins, were found only in the family Bacillaceae, or even in a subset of Bacillus spp. Phylogenetic profiles of sporulation genes, compiled in this work, confirm the presence of a common sporulation gene core, but also illuminate the diversity of the sporulation processes within various lineages. These profiles should help further experimental studies of uncharacterized widespread sporulation genes, which would ultimately allow delineation of the minimal set(s) of sporulation-specific genes in Bacilli and Clostridia. Published 2012. This article is a U.S. Government work and is in the public domain in the USA.

[Accurate diagnosis of Pseudomonas luteola in routine microbiology laboratory: on the occasion of two isolates].

PubMed

Çiçek, Muharrem; Hasçelik, Gülşen; Müştak, H Kaan; Diker, K Serdar; Şener, Burçin

2016-10-01

Pseudomonas luteola which was previously known as Chryseomonas luteola; is a gram-negative, non-fermentative, aerobic, motile, non-spore-forming bacillus. It is frequently found as a saprophyte in soil, water and other damp environments and is an opportunistic pathogen in patients with underlying medical disorders or with indwelling catheters. It has been reported as an uncommon cause of bacteremia, sepsis, septic arthritis, meningitis, endocarditis, and peritonitis. Thus, early and accurate identification of this rare species is important for the treatment and also to provide information about the epidemiology of P.luteola infections. This report was aimed to draw attention to the accurate identification of P.luteola in clinical samples, upon the isolation and identification in two cases in the medical microbiology laboratory of a university hospital. In February 2011, a 66-year-old man, with chronic obstructive pulmonary disease, coronary artery disease and aplastic anemia, was admitted to our hospital due to progressive dyspnea. A chest tube was inserted on the 20th day of admission by the reason of recurrent pleural effusion. Staphylococcus aureus and a non-fermentative gram-negative bacillus (NFGNB) with wrinkled, sticky yellow colonies were isolated from the pleural fluid sample obtained on the 9th day following the insertion of the chest tube. In February 2012, a 7-year-old male cystic fibrosis patient who had no signs and symptoms of acute pulmonary exacerbation was admitted to the hospital for a routine control. This patient had chronic colonization with Pseudomonas aeruginosa and S.aureus and his sputum sample obtained at this visit revealed isolation of P.aeruginosa, S.aureus, Aspergillus fumigatus and a wrinkled, sticky yellow NFGNB. Both of these NFGNB were identified as P.luteola by the Phoenix automated microbial identification system (BD Diagnostics, USA). To evaluate the microbiological characteristics of these two isolates, the strains were further analysed by VITEK MS (bioMerieux, France) and Microflex LT mass spectrometer (Bruker Daltonics, Germany). Both of the MALDI-TOF-MS systems identified the isolates as P.luteola and 16S rRNA gene sequencing (ABI PRISM 3100, Applied Biosystems, USA) also confirmed the identification. The strains had wrinkled, sticky yellow colonies which were oxidase-negative, catalase-positive and non-fermentative. The Gram stained smears of the colonies revealed clusters of gram-negative bacilli probably embedded into a biofilm matrix. Since there are no accepted standards for testing the antibiotic susceptibility of P.luteola strains, the standards determined by CLSI for "other non-Enterobacteriaceae" (non-fermentative bacteria excluding P.aeruginosa, Acinetobacter spp., Burkholderia cepacia, B.mallei, B.pseudomallei and Stenotrophomonas maltophilia) were used for the susceptibility testing. Gradient MIC method (E-Test, bioMerieux, France) revealed that the isolates were susceptible to gentamicin, piperacillin-tazobactam, ceftazidime, cefepime, meropenem, colistin and levofloxacin. Accurate and prompt identification of P.luteola which is identified as a rare pathogen in serious cases is of critical importance since it has been suggested that this organism is likely to become more frequent as a nosocomial pathogen since the interventional processes increase in current medical practice. This report supported that Phoenix automated phenotypic identification system (BD Diagnostics, USA) and the two MALDI-TOF-MS based systems (VITEK MS and Bruker Microflex LT mass spectrometer) were successfull in the accurate identification of P.luteola.

Novel pharmacotherapy for the treatment of hospital-acquired and ventilator-associated pneumonia caused by resistant gram-negative bacteria.

PubMed

Kidd, James M; Kuti, Joseph L; Nicolau, David P

2018-03-01

Hospital-acquired and ventilator-associated bacterial pneumonia (HABP/VABP) are among the most prevalent infections in hospitalized patients, particularly those in the intensive care unit. Importantly, the frequency of multidrug resistant (MDR) Gram-negative (GN) bacteria as the bacteriologic cause of HABP/VABP is increasing. These include MDR Pseudomonas aeruginosa, Acinetobacter baumannii, and carbapenem resistant Enterobacteriaceae (CRE). Few antibiotics are currently available when such MDR Gram-negatives are encountered and older agents such as polymyxin B, colistin (polymyxin E), and tigecycline have typically performed poorly in HABP/VABP. Areas covered: In this review, the authors summarize novel antibiotics which have reached phase 3 clinical trials including patients with HABP/VABP. For each agent, the spectrum of activity, pertinent pharmacological characteristics, clinical trial data, and potential utility in the treatment of MDR-GN HABP/VABP is discussed. Expert opinion: Novel antibiotics currently available, and those soon to be, will expand opportunities to treat HABP/VABP caused by MDR-GN organisms and minimize the use of more toxic, less effective drugs. However, with sparse clinical data available, defining the appropriate role for each of the new agents is challenging. In order to maximize the utility of these antibiotics, combination therapy and the role of therapeutic drug monitoring should be investigated.

Gram-negative urinary tract infections and increasing isolation of ESBL-producing or ceftazidime-resistant strains in children: results from a single-centre survey.

PubMed

Giardino, Stefano; Bandettini, Roberto; Perotti, Maddalena; Piaggio, Giorgio; Degl'Innocenti, Ludovica; Sacco, Rosanna; Ciucci, Antonella; Caviglia, Ilaria; Barabino, Paola; Ginocchio, Francesca; Losurdo, Giuseppe; Haupt, Riccardo; Castagnola, Elio

2013-03-01

Urinary tract infections (UTIs) are an important cause of morbidity in paediatrics, especially related to urinary tract malformation and neurogenic bladder dysfunction. The infection control team of Istituto Giannina Gaslini, Genova, Italy, performs a prospective survey on the epidemiology of UTI in children admitted in the hospital, and data are expressed as episodes/1000 days of hospital admission. From 2007 to 2011 there was an increase in the rate of Gram-negative UTIs, especially in the Nephrology Unit (from 11.63 to 27.48, r-coefficient 0.95, P minor 0.05), associated with an increase in infections due to ESBL-producing strains (from 0.54 to 2.55, r-coefficient 0.89, P 0.05). This study indicates that there is an increase in the rate of Gram-negative UTIs, also due to resistant strains. The cause may be multifactorial, but it is noteworthy that it has been mainly observed in a ward where low-dose, long-term administration of antibacterial prophylaxis in children with urinary malformations or neurogenic bladder dysfunction is routine. This phenomenon gives cause for concern and should be monitored carefully to avoid the risk of selecting resistant bacteria that have no therapeutic options.

Chemical Composition and Antibacterial Activity of Essential Oil of Cosmos bipinnatus Cav. Leaves from South Africa

PubMed Central

Olajuyigbe, Olufunmiso; Ashafa, Anofi

2014-01-01

The chemical composition of essential oils isolated from the leaves of Cosmos bipinnatus and its antibacterial activity were analyzed by GC-MS and microbroth dilution assay respectively. The essential oil extracted from this plant was predominantly composed of monoterpenes (69.62%) and sesquiterpenes (22.73%). The antibacterial assay showed that the oil had significant inhibitory effects against both Gram-negative and Gram-positive bacteria isolates. The MIC of Gram-positive strains ranged between 0.16 and 0.31 mg/mL while those of Gram-negative bacteria ranged between 0.31 and 0.63 mg/mL. The Gram-positive bacteria were more susceptible to the essential oil than the Gram-negative bacteria. Most of the major components of this oil in other plants have been reported for antimicrobial activities. The antibacterial activity can be attributed to effects of the combination of several components of the oil. The results indicate that the C. bipinnatus might be exploited as natural antibacterial agent and have application in the treatment of several infectious diseases caused by these bacteria. Since this species is endemic to the eastern Free State, the plant could be collected during its bloom and used efficiently in the management of bacterial infections in South Africa. PMID:25587332

Chemical Composition and Antibacterial Activity of Essential Oil of Cosmos bipinnatus Cav. Leaves from South Africa.

PubMed

Olajuyigbe, Olufunmiso; Ashafa, Anofi

2014-01-01

The chemical composition of essential oils isolated from the leaves of Cosmos bipinnatus and its antibacterial activity were analyzed by GC-MS and microbroth dilution assay respectively. The essential oil extracted from this plant was predominantly composed of monoterpenes (69.62%) and sesquiterpenes (22.73%). The antibacterial assay showed that the oil had significant inhibitory effects against both Gram-negative and Gram-positive bacteria isolates. The MIC of Gram-positive strains ranged between 0.16 and 0.31 mg/mL while those of Gram-negative bacteria ranged between 0.31 and 0.63 mg/mL. The Gram-positive bacteria were more susceptible to the essential oil than the Gram-negative bacteria. Most of the major components of this oil in other plants have been reported for antimicrobial activities. The antibacterial activity can be attributed to effects of the combination of several components of the oil. The results indicate that the C. bipinnatus might be exploited as natural antibacterial agent and have application in the treatment of several infectious diseases caused by these bacteria. Since this species is endemic to the eastern Free State, the plant could be collected during its bloom and used efficiently in the management of bacterial infections in South Africa.

A Case Series and Review of Bacillus Cereus Endocarditis from India.

PubMed

Gopinathan, Anusha; Kumar, Anil; Sen, Amitabh C; Sudha, Srisruthy; Varma, Praveen; Gs, Sunil; Eapen, Malini; Dinesh, Kavitha R

2018-01-01

Bacillus cereus is a gram positive bacilli found commonly in the soil and environment. It is a bacteria rarely associated with endocarditis. Intravenous drug abuse, presence of valvular defects, pacemakers, immunodeficiency are some of the known risk factors for B.cereus endocarditis. We present here a case series of two patients with B.cereus endocarditis along with a review of the literature. This is the first report of B.cereus endocarditis from India to the best of our knowledge.

Clostridium perfringens type A fatal acute hemorrhagic gastroenteritis in a dog.

PubMed

Schlegel, Ben J; Van Dreumel, Tony; Slavić, Durda; Prescott, John F

2012-05-01

The morning after participating in a dog show, a 2-year-old Pomeranian dog was found dead in a pool of bloody feces. Necropsy revealed hemorrhagic gastroenteritis of the entire gastrointestinal tract, with many Gram-positive bacilli on the surface and in the lumen and crypts of the intestine. Enterotoxin-positive type A Clostridium perfringens were isolated in large numbers. This dramatic case of fatal C. perfringens gastroenteritis highlights the need to better understand the role of this bacterium in enteric disease of dogs.

Tepidibacillus decaturensis sp. nov., a microaerophilic, moderately thermophilic iron-reducing bacterium isolated from 1.7 km depth groundwater

DOE PAGES

Dong, Yiran; Sanford, Robert A.; Boyanov, Maxim I.; ...

2016-10-01

Here, a Gram-stain-negative, microaerophilic rod-shaped organism designated as strain Z9 T was isolated from groundwater of 1.7 km depth from the Mt. Simon Sandstone of the Illinois Basin, Illinois, USA. Cells of strain Z9 T were rod shaped with dimensions of 0.3×(1–10) µm and stained Gram-negative. Strain Z9 T grew within the temperature range 20–60 °C (optimum at 30–40 °C), between pH 5 and 8 (optimum 5.2–5.8) and under salt concentrations of 1–5 % (w/v) NaCl (optimum 2.5 % NaCl). In addition to growth by fermentation and nitrate reduction, this strain was able to reduce Fe(III), Mn(IV), Co(III) and Cr(VI)more » when H 2 or organic carbon was available as the electron donor, but did not actively reduce oxidized sulfur compounds (e.g. sulfate, thiosulfate or S 0). The G+C content of the DNA from strain Z9 T was 36.1 mol%. Phylogenetic analysis of the 16S rRNA gene from strain Z9 T showed that it belongs to the class Bacilli and shares 97 % sequence similarity with the only currently characterized member of the genus Tepidibacillus, T. fermentans. Based on the physiological distinctness and phylogenetic information, strain Z9 T represents a novel species within the genus Tepidibacillus, for which the name Tepidibacillus decaturensis sp. nov. is proposed. The type strain is Z9 T (=ATCC BAA-2644 T=DSM 103037 T).« less

Tepidibacillus decaturensis sp. nov., a microaerophilic, moderately thermophilic iron-reducing bacterium isolated from 1.7 km depth groundwater

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dong, Yiran; Sanford, Robert A.; Boyanov, Maxim I.

Here, a Gram-stain-negative, microaerophilic rod-shaped organism designated as strain Z9 T was isolated from groundwater of 1.7 km depth from the Mt. Simon Sandstone of the Illinois Basin, Illinois, USA. Cells of strain Z9 T were rod shaped with dimensions of 0.3×(1–10) µm and stained Gram-negative. Strain Z9 T grew within the temperature range 20–60 °C (optimum at 30–40 °C), between pH 5 and 8 (optimum 5.2–5.8) and under salt concentrations of 1–5 % (w/v) NaCl (optimum 2.5 % NaCl). In addition to growth by fermentation and nitrate reduction, this strain was able to reduce Fe(III), Mn(IV), Co(III) and Cr(VI)more » when H 2 or organic carbon was available as the electron donor, but did not actively reduce oxidized sulfur compounds (e.g. sulfate, thiosulfate or S 0). The G+C content of the DNA from strain Z9 T was 36.1 mol%. Phylogenetic analysis of the 16S rRNA gene from strain Z9 T showed that it belongs to the class Bacilli and shares 97 % sequence similarity with the only currently characterized member of the genus Tepidibacillus, T. fermentans. Based on the physiological distinctness and phylogenetic information, strain Z9 T represents a novel species within the genus Tepidibacillus, for which the name Tepidibacillus decaturensis sp. nov. is proposed. The type strain is Z9 T (=ATCC BAA-2644 T=DSM 103037 T).« less

[Isolation of heterotrophic nitrifiers/aerobic denitrifiers and their roles in N2O production for different incubations].

PubMed

Jiang, Jing-Yan; Hu, Zheng-Hua; Huang, Yao

2009-07-15

Soil microorganisms are important sources of N2O for the atmosphere. Peak emissions of N2O are often observed after wetting of soil. The simultaneous heterotrophic nitrifying and aerobic denitrifying bacteria with respect to N2O emission were studied to obtain more information about the microbiologcal aspects of peak emissions. Using acetamide as the C and N source, two strains of nitrifying and denitrifying bacteria were isolated, coded as XM1 and HX2,respectively. XM1 strain was Gram-negative chain-like bacilli, and the HX2 was Gram-negative cocci. In enrichment culture, N2O production of HX2 was 76 times more than XM1. Two strains could grow with glucose, mannitol or sodium tartrate as sole carbon source, respectively. They could nitrify with sodium nitrate or denitrify with ammonium sulfate as unique nitrogen source, and produce intermediate product nitrite. XM1 strain growth velocity and nitrite formation were obviously higher than HX2. The phylogentic analysis based on partial 16S rDNA showed that two isolated strains were the closest relative of Pseudomonas sp.99% sequence similarity. Under different WFPS (water-filled-pore-space) conditions, the aerobic autoclaved soil incubation trial showed that, HX2 strain was suitable for growing in 30% WFPS, and N2O production was (36.01 +/- 2.48) ng/g which was 1.9 times than that in 60% WFPS. But XM1 was suitable for growing in 60% WFPS and almost had no N2O production. To investigate the nitrifying and denitrifying mechanisms of heterotrophic nitrifiers/aerobic denitrifiers should be useful for mastering the mitigation way of soil N2O emission in future.

Validation of the Zürich burn-biofilm model.

PubMed

Guggenheim, Merlin; Thurnheer, Thomas; Gmür, Rudolf; Giovanoli, Pietro; Guggenheim, Bernhard

2011-11-01

Despite advances in the use of topical and parenteral antimicrobial therapy and the practice of early tangential burn-wound excision, bacterial infection remains a major problem in the management of burn victims today. The purpose of this study was to design and evaluate a polyspecies biofilm model with bacteria known to cause severe infections in burn patients. The model is simple to prepare, maintain and analyse, and allows for short-term exposure to antimicrobials. Initial experiments showed that it was impossible to establish balanced polyspecies biofilms with an inoculum of Gram-positive and -negative bacteria. After 64.5 h of incubation, the Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) had suppressed the Gram-positives (Enterococcus faecalis, Staphylococcus aureus and Streptococcus intermedius). However, adding the Gram-negative bacteria after 41.5 h to an established biofilm of Gram-positives resulted in a balanced microbial consortium. After 64.5 h, all species were present in high numbers (10(7) to 10(8) colony forming units (CFU) per biofilm). Multiple repetitions showed high reproducibility of biofilm formation without significant differences between and within experiments. Combined fluorescence in situ hybridisation/confocal laser scanning microscopy (FISH/CLSM) analyses, for which biofilms had to be grown on a different non-flexible substrate (hydroxy apatite), revealed that, by 41.5 h, the biofilm consisted of an almost confluent layer of bacteria firmly adherent to the substratum. After 64.5 h (22 h after the addition of the Gram negatives), the biofilm consisted of a confluent mixture of single cells, an abundance of galaxies of bacteria with small lacunae and large amounts of extracellular matrix polysaccharides. The polyspecies biofilm model contains the most prevalent burn-associated Gram-positive and Gram-negative bacterial pathogens and mimics the Gram-negative shift observed in vivo. It shows excellent reproducibility. It should allow adaptation to the bacterial spectrum prevalent in different burn centres and lead to a much more reliable investigation of the efficiency of topical antimicrobial agents than models operating with planktonic bacteria. The experiments further open up the perspective to create an in vivo model using these biofilms as infectious agents. Copyright © 2011 Elsevier Ltd and ISBI. All rights reserved.

Bacterial Viability within Dental Calculus: An Untrodden, Inquisitive Clinico-Patho- Microbiological Research.

PubMed

Gupta, Swati; Jain, P K; Kumra, Madhumani; Rehani, Shweta; Mathias, Yulia; Gupta, Ramakant; Mehendiratta, Monica; Chander, Anil

2016-07-01

Chronic inflammatory periodontal diseases i.e. gingivitis and periodontitis are one of the most common afflictions faced by human beings. Dental plaque, which is a pool of pathogenic microorganisms, remains to be current mainstay in etiopathogenesis. Dental calculus, which is a mineralized product of this plaque remains ignored and is considered merely as an ash heap of minor significance. However, the intriguing array in disease etiopathogenesis bulldozed researchers to suspect the role of calculus in disease chrysalis but still the viability of bacteria inside calculus and thus its pathogenicity remains an intricacy; the answer to which lies in the Pandora's Box. The present study was undertaken to investigate the viability of bacteria within dental calculus along with their identification. Also, to classify dental calculus on the basis of mineralization and to observe the variation of viable microflora found in dental calculus with the extent of mineralization and disease severity. A total of 60 samples were obtained, by harvesting two samples of supragingival calculus from each patient having chronic inflammatory periodontal disease. These samples were divided into two groups (Group A and Group B). Samples of Group A were kept non-irradiated and samples of Group B were exposed to UV radiation. The samples were categorized into less, moderately and highly mineralized according to the force required for crushing them. All the crushed calculus samples were then divided into three parts. These were used for dark-field microscopy, gram staining and bacterial cultures. Bacterial identification of the cultures obtained was also carried out by performing various biochemical assays. The present study revealed the presence of motile spirochaetes within the samples under dark-field microscope. Gram staining revealed presence of numerous gram positive cocci and gram negative bacilli. Bacterial cultures showed growth of variety of aerobic and capnophilic microorganisms. The present study concludes the presence of viable aerobic and capnophilic bacteria inside dental calculus which may reside within the lacunae and channels in the calculus.

Microbiology and management of joint and bone infections due to anaerobic bacteria.

PubMed

Brook, Itzhak

2008-03-01

To describes the microbiology, diagnosis, and management of septic arthritis and osteomyelitis due to anaerobic bacteria. The predominant anaerobes in arthritis are anaerobic Gram-negative bacilli (AGNB) including the Bacteroides fragilis group, Fusobacterium spp., Peptostreptococcus spp., and Propionibacterium acnes. Infection with P. acnes is associated with a prosthetic joint, previous surgery, and trauma. B. fragilis group is associated with distant infection, Clostridium spp. with trauma, and Fusobacterium spp. with oropharyngeal infection. Most cases of anaerobic arthritis, in contrast to anaerobic osteomyelitis, involved a single isolate, and most cases are secondary to hematogenous spread. The predominant anaerobes in osteomyelitis are Bacteroides, Peptostreptococcus, Fusobacterium, and Clostridium spp. as well as P. acnes. Conditions predisposing to bone infections are vascular disease, bites, contiguous infection, peripheral neuropathy, hematogenous spread, and trauma. Pigmented Prevotella and Porphyromonas spp. are mostly isolated in skull and bite infections, members of the B. fragilis group in hand and feet infections, and Fusobacterium spp. in skull, bite, and hematogenous long bone infections. Many patients with osteomyelitis due to anaerobic bacteria have evidence of an anaerobic infection elsewhere in the body that is the source of the organisms involved in the osteomyelitis. Treatment of arthritis and osteomyelitis involving anaerobic bacteria includes symptomatic therapy, immobilization in some cases, adequate drainage of purulent material, and antibiotic therapy effective against these organisms. Anaerobic bacteria can cause septic arthritis and osteomyelitis. Correct diagnosis and appropriate therapy are important contributor to successful outcome.

Anaerobic Bacteria in Clinical Specimens – Frequent, But a Neglected Lot: A Five Year Experience at a Tertiary Care Hospital

PubMed Central

Shenoy, Padmaja Ananth; Gawda, Ashwini; Shetty, Seema; Anegundi, Renuka; Varma, Muralidhar; Mukhopadhyay, Chiranjay; Chawla, Kiran

2017-01-01

Introduction Anaerobic bacteria which constitute a significant proportion of the normal microbiota also cause variety of infections involving various anatomic sites. Considering the tedious culture techniques with longer turnaround time, anaerobic cultures are usually neglected by clinicians and microbiologists. Aim To study the frequency of isolation of different anaerobic bacteria from various clinical specimens. Materials and Methods A retrospective study to analyse the frequency of isolation of different anaerobic bacteria, was conducted over a period of five years from 2011 to 2015 including various clinical specimens submitted to anaerobic division of Microbiology laboratory. Anaerobic bacteria were isolated and identified following standard bacteriological techniques. Results Pathogenic anaerobes (n=336) were isolated from 278 (12.48%) of overall 2227 specimens processed with an average yield of 1.2 isolates. Anaerobes were isolated as polymicrobial flora with or without aerobic bacterial pathogens in 159 (57.2%) patients. Anaerobic Gram-negative bacilli (140, 41.7%) were the predominant isolates. B. fragilis group (67, 19.9%) were the most commonly isolated anaerobic pathogens. Anaerobes were predominantly isolated from deep seated abscess (23.9%). Conclusion Pathogenic anaerobes were isolated from various infection sites. Unless culture and susceptibility tests are performed as a routine, true magnitude of antimicrobial resistance among anaerobic pathogens will not be known. Knowledge of the distribution of these organisms may assist in the selection of appropriate empirical therapy for anaerobic infections. PMID:28892897

Skin diseases during floods in Thailand.

PubMed

Vachiramon, Vasanop; Busaracome, Ploysyne; Chongtrakool, Piriyaporn; Puavilai, Siripen

2008-04-01

Floods are natural disasters that occur occasionally in Thailand. The most common form skin diseases due to floods are infectious dermatoses especially superficial fungal infection. However the microbiologic evidences have not been evaluated. To evaluate the most common skin diseases during floods and identify the organism that causes skin maceration at web space(s) of toes (Hong Kong foot). Patients who complained of skin problems were evaluated at the temporary outpatient clinic during October 2006. Skin specimens from all patients who had itches and skin maceration at web space(s) of toes were cultured. Ninety-six patients were evaluated (38 males and 58 females). Eczema was the most prevalent dermatosis, which accounted for 34.5% of the total skin problems and the great majority of these cases were irritant contact dermatitis. Sixteen cases presented with itch and skin maceration at web space(s) of toes. All of them were colonized with various microorganisms. Gram-negative bacilli were the most prevalent ones and were found in 14 out of 16 specimens. Fungal culture was positive in only two specimens. Eczema is the most common dermatosis during floods. Skin maceration at web space(s) of toes, which were thought to be fungal infection, are chronic irritant dermatitis with secondary bacterial colonization. Only a few cases were fungal infection. Microbiologic investigation should be done in these patients. Unfortunately, it is not practical in such a situation. Topical medications that have the combination of antiinflammatory, antibacterial and antifungal properties are the most suitable medications.

Carbapenem-Resistant Enterobacteriaceae Transmission in Health Care Facilities - Wisconsin, February-May 2015.

PubMed

Elbadawi, Lina I; Borlaug, Gwen; Gundlach, Kristin M; Monson, Timothy; Warshauer, David; Walters, Maroya S; Kallen, Alexander; Gulvik, Christopher A; Davis, Jeffrey P

2016-09-02

Carbapenem-resistant Enterobacteriaceae (CRE) are multidrug-resistant gram-negative bacilli that can cause infections associated with high case fatality rates, and are emerging as epidemiologically important health care-associated pathogens in the United States (1). Prevention of CRE transmission in health care settings is dependent on recognition of cases, isolation of colonized and infected patients, effective use of infection control measures, and the correct use of antibiotics. The use of molecular technologies, including polymerase chain reaction (PCR) testing, pulsed-field gel electrophoresis (PFGE), and whole genome sequencing (WGS), can lead to detection of transmission events and interruption of transmission. In Wisconsin, acute care and critical access hospitals report laboratory-identified CRE to the Wisconsin Division of Public Health (WDPH), and clinical laboratories submit CRE isolates to the Wisconsin State Laboratory of Hygiene (WSLH) for molecular testing. During February-May 2015, a total of 49 CRE isolates from 46 patients were submitted to WSLH. On June 8, WSLH informed WDPH of five carbapenemase-producing CRE isolates with closely related PFGE patterns identified among four inpatients at two hospitals in southeastern Wisconsin. An investigation revealed a high degree of genetic relatedness among the patients' isolates, but did not identify the mechanism of transmission between the two facilities. No breaches in recommended practices were identified; after reviewing respiratory care procedures, no further cases were identified. Routine hospital- and laboratory-based surveillance can detect and prevent health care transmission of CRE.

Risk factors for infection with multidrug-resistant bacteria in non-ventilated patients with hospital-acquired pneumonia.

PubMed

Seligman, Renato; Ramos-Lima, Luis Francisco; Oliveira, Vivian do Amaral; Sanvicente, Carina; Sartori, Juliana; Pacheco, Elyara Fiorin

2013-01-01

To identify risk factors for the development of hospital-acquired pneumonia (HAP) caused by multidrug-resistant (MDR) bacteria in non-ventilated patients. This was a retrospective observational cohort study conducted over a three-year period at a tertiary-care teaching hospital. We included only non-ventilated patients diagnosed with HAP and presenting with positive bacterial cultures. Categorical variables were compared with chi-square test. Logistic regression analysis was used to determine risk factors for HAP caused by MDR bacteria. Of the 140 patients diagnosed with HAP, 59 (42.1%) were infected with MDR strains. Among the patients infected with methicillin-resistant Staphylococcus aureus and those infected with methicillin-susceptible S. aureus, mortality was 45.9% and 50.0%, respectively (p = 0.763). Among the patients infected with MDR and those infected with non-MDR gram-negative bacilli, mortality was 45.8% and 38.3%, respectively (p = 0.527). Univariate analysis identified the following risk factors for infection with MDR bacteria: COPD; congestive heart failure; chronic renal failure; dialysis; urinary catheterization; extrapulmonary infection; and use of antimicrobial therapy within the last 10 days before the diagnosis of HAP. Multivariate analysis showed that the use of antibiotics within the last 10 days before the diagnosis of HAP was the only independent predictor of infection with MDR bacteria (OR = 3.45; 95% CI: 1.56-7.61; p = 0.002). In this single-center study, the use of broad-spectrum antibiotics within the last 10 days before the diagnosis of HAP was the only independent predictor of infection with MDR bacteria in non-ventilated patients with HAP.

Risk factors for infection with multidrug-resistant bacteria in non-ventilated patients with hospital-acquired pneumonia*,**

PubMed Central

Seligman, Renato; Ramos-Lima, Luis Francisco; Oliveira, Vivian do Amaral; Sanvicente, Carina; Sartori, Juliana; Pacheco, Elyara Fiorin

2013-01-01

OBJECTIVE: To identify risk factors for the development of hospital-acquired pneumonia (HAP) caused by multidrug-resistant (MDR) bacteria in non-ventilated patients. METHODS: This was a retrospective observational cohort study conducted over a three-year period at a tertiary-care teaching hospital. We included only non-ventilated patients diagnosed with HAP and presenting with positive bacterial cultures. Categorical variables were compared with chi-square test. Logistic regression analysis was used to determine risk factors for HAP caused by MDR bacteria. RESULTS: Of the 140 patients diagnosed with HAP, 59 (42.1%) were infected with MDR strains. Among the patients infected with methicillin-resistant Staphylococcus aureus and those infected with methicillin-susceptible S. aureus, mortality was 45.9% and 50.0%, respectively (p = 0.763). Among the patients infected with MDR and those infected with non-MDR gram-negative bacilli, mortality was 45.8% and 38.3%, respectively (p = 0.527). Univariate analysis identified the following risk factors for infection with MDR bacteria: COPD; congestive heart failure; chronic renal failure; dialysis; urinary catheterization; extrapulmonary infection; and use of antimicrobial therapy within the last 10 days before the diagnosis of HAP. Multivariate analysis showed that the use of antibiotics within the last 10 days before the diagnosis of HAP was the only independent predictor of infection with MDR bacteria (OR = 3.45; 95% CI: 1.56-7.61; p = 0.002). CONCLUSIONS: In this single-center study, the use of broad-spectrum antibiotics within the last 10 days before the diagnosis of HAP was the only independent predictor of infection with MDR bacteria in non-ventilated patients with HAP. PMID:23857697

Burkholderia mallei and Burkholderia pseudomallei stimulate differential inflammatory responses from human alveolar type II cells (ATII) and macrophages.

PubMed

Lu, Richard; Popov, Vsevolod; Patel, Jignesh; Eaves-Pyles, Tonyia

2012-01-01

Alveolar type II pneumocytes (ATII) and alveolar macrophages (AM) play a crucial role in the lung's innate immune response. Burkholderia pseudomallei (BP) and Burkholderia mallei (BM) are facultative Gram-negative bacilli that cause melioidosis and glanders, respectively. The inhalation of these pathogens can cause lethal disease and death in humans. We sought to compare the pathogenesis of and host responses to BP and BM through contact with human primary ATII cells and monocytes-derived macrophages (MDM). We hypothesized that because BP and BM induce different disease outcomes, each pathogen would induce distinct, unique host immune responses from resident pulmonary cells. Our findings showed that BP adhered readily to ATII cells compared to BM. BP, but not BM, was rapidly internalized by macrophages where it replicated to high numbers. Further, BP-induced significantly higher levels of pro-inflammatory cytokine secretion from ATII cells (IL-6, IL-8) and macrophages (IL-6, TNFα) at 6 h post-infection compared to BM (p < 0.05). Interestingly, BM-induced the anti-inflammatory cytokine, IL-10, in ATII cells and macrophages at 6 h post-infection, with delayed induction of inflammatory cytokines at 24 h post-infection. Because BP is flagellated and produces LPS, we confirmed that it stimulated both Toll-like receptor (TLR) 4 and TLR5 via NF-κb activation while the non-flagellated BM stimulated only TLR4. These data show the differences in BP and BM pathogenicity in the lung when infecting human ATII cells and macrophages and demonstrate the ability of these pathogens to elicit distinct immune responses from resident lung cells which may open new targets for therapeutic intervention to fight against these pathogens.

Pharmacokinetic-Pharmacodynamic-Model-Guided Doripenem Dosing in Critically Ill Patients▿ †

PubMed Central

Samtani, Mahesh N.; Flamm, Robert; Kaniga, Koné; Nandy, Partha

2010-01-01

The growing number of infections caused by multidrug-resistant pathogens has prompted a more rational use of available antibiotics given the paucity of new, effective agents. Monte Carlo simulations were utilized to determine the appropriateness of several doripenem dosing regimens based on the probability of attaining the critical drug exposure metric of time that drug concentrations remain above the drug MIC (T>MIC) for 35% (and lower thresholds) of the dosing interval in >80 to 90% of the population (T>MIC 35% target). This exposure level generally correlates with in vivo efficacy for carbapenems. In patients with creatinine clearance of >50 ml/min, a 500-mg dose of doripenem infused over 1 h every 8 h is expected to be effective against bacilli with doripenem MICs of ≤1 μg/ml based on a T>MIC 35% target and MICs of ≤2 μg/ml based on lower targets. A longer, 4-hour infusion time improved target attainment in most cases, such that the T>MIC was adequate for pathogens with doripenem MICs as high as 4 μg/ml. Efficacy is expected for infections caused by pathogens with doripenem MICs of ≤2 μg/ml in patients with moderate renal impairment (creatinine clearance, 30 to 50 ml/min) who receive doripenem at 250 mg infused over 1 h every 8 h and in patients with severe impairment (creatinine clearance between 10 and 29 ml/min) who receive doripenem at 250 mg, infused over 1 h or 4 h, every 12 h. Results of pharmacokinetics/pharmacodynamics (PK/PD) modeling can guide dose optimization, thereby potentially increasing the clinical efficacy of doripenem against serious Gram-negative bacterial infections. PMID:20385857

Health care workers' mobile phones: a potential cause of microbial cross-contamination between hospitals and community.

PubMed

Ustun, Cemal; Cihangiroglu, Mustafa

2012-01-01

This study evaluated the microbial contamination of health care workers' (HCWs) mobile phones. The study was conducted at a secondary referral hospital in July 2010. Samples were taken from all surfaces of the mobile phones using a sterile swab, and incubated on Brain Heart Infusion agar at 37.5°C for 24 hr. Any isolated microorganisms were grown aerobically on 5% sheep blood agar and eosin methylene-blue agar medium at 37.5°C for 24-48 hr. The Sceptor microdilution system was used to identify the microorganisms, together with conventional methods. The oxacillin disc diffusion test and double-disc synergy test were used to identify methicillin-resistant Staphylococcus aureus (MRSA) and expanded-spectrum beta-lactamase (ESBL)-producing Gram-negative bacilli, respectively. The mobile phones were also categorized according to whether the HCWs used them in the intensive care unit (ICU). Overall, 183 mobile phones were screened: 94 (51.4%) from nurses, 32 (17.5%) from laboratory workers, and 57 (31.1%) from health care staff. In total, 179 (97.8%) culture-positive specimens were isolated from the 183 mobile phones, including 17 (9.5%) MRSA and 20 (11.2%) ESBL-producing Escherichia coli, which can cause nosocomial infections. No statistical difference was observed in the recovery of MRSA (p = 0.3) and ESBL-producing E. coli (p = 0.6) between the HCW groups. Forty-four (24.6%) of the 179 specimens were isolated from mobile phones of ICU workers, including two MRSA and nine ESBL-producing E. coli. A significant (p = 0.02) difference was detected in the isolation of ESBL-producing E. coli between ICU workers and non-ICU workers. HCWs' mobile phones are potential vectors for transferring nosocomial pathogens between HCWs, patients, and the community.

Effects of pathogen-specific clinical mastitis on probability of conception in Holstein dairy cows.

PubMed

Hertl, J A; Schukken, Y H; Welcome, F L; Tauer, L W; Gröhn, Y T

2014-11-01

The objective of this study was to estimate the effects of pathogen-specific clinical mastitis (CM), occurring in different weekly intervals before or after artificial insemination (AI), on the probability of conception in Holstein cows. Clinical mastitis occurring in weekly intervals from 6 wk before until 6 wk after AI was modeled. The first 4 AI in a cow's lactation were included. The following categories of pathogens were studied: Streptococcus spp. (comprising Streptococcus dysgalactiae, Streptococcus uberis, and other Streptococcus spp.); Staphylococcus aureus; coagulase-negative staphylococci (CNS); Escherichia coli; Klebsiella spp.; cases with CM signs but no bacterial growth (above the level that can be detected from our microbiological procedures) observed in the culture sample and cases with contamination (≥ 3 pathogens in the sample); and other pathogens [including Citrobacter, yeasts, Trueperella pyogenes, gram-negative bacilli (i.e., gram-negative organisms other than E. coli, Klebsiella spp., Enterobacter, and Citrobacter), Corynebacterium bovis, Corynebacterium spp., Pasteurella, Enterococcus, Pseudomonas, Mycoplasma, Prototheca, and others]. Other factors included in the model were parity (1, 2, 3, 4 and higher), season of AI (winter, spring, summer, autumn), day in lactation of first AI, farm, and other non-CM diseases (retained placenta, metritis, ketosis, displaced abomasum). Data from 90,271 AI in 39,361 lactations in 20,328 cows collected from 2003/2004 to 2011 from 5 New York State dairy farms were analyzed in a generalized linear mixed model with a Poisson distribution. The largest reductions in probability of conception were associated with CM occurring in the week before AI or in the 2 wk following AI. Escherichia coli and Klebsiella spp. had the greatest adverse effects on probability of conception. The probability of conception for a cow with any combination of characteristics may be calculated based on the parameter estimates. These findings may be helpful to farmers in assessing reproduction in their dairy cows for more effective cow management. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Bloodstream infections in patients with hematological malignancies: which is more fatal – cancer or resistant pathogens?

PubMed Central

Gedik, Habip; Şimşek, Funda; Kantürk, Arzu; Yildirmak, Taner; Arica, Deniz; Aydin, Demet; Demirel, Naciye; Yokuş, Osman

2014-01-01

Background The primary objective of this study was to report the incidence of bloodstream infections (BSIs) and clinically or microbiologically proven bacterial or fungal BSIs during neutropenic episodes in patients with hematological malignancies. Methods In this retrospective observational study, all patients in the hematology department older than 14 years who developed febrile neutropenia during chemotherapy for hematological cancers were evaluated. Patients were included if they had experienced at least one neutropenic episode between November 2010 and November 2012 due to chemotherapy in the hematology ward. Results During 282 febrile episodes in 126 patients, 66 (23%) episodes of bacteremia and 24 (8%) episodes of fungemia were recorded in 48 (38%) and 18 (14%) patients, respectively. Gram-negative bacteria caused 74% (n=49) of all bacteremic episodes. Carbapenem-resistant Gram-negative bacteria (n=6) caused 12% and 9% of Gram-negative bacteremia episodes and all bacteremia episodes, respectively. Carbapenem-resistant Gram-negative bacteria included Acinetobacter baumannii (n=4), Pseudomonas aeruginosa (n=1), and Serratia marcescens (n=1). Culture-proven invasive fungal infection occurred in 24 episodes in 18 cases during the study period, with 15 episodes in ten cases occurring in the first study year and nine episodes in eight cases in the second study year. In 13 of 18 cases (72%) with bloodstream yeast infections, previous azole exposure was recorded. Candida parapsilosis, C. glabrata, and C. albicans isolates were resistant to voriconazole and fluconazole. Conclusion BSIs that occur during febrile neutropenic episodes in hematology patients due to Gram-negative bacteria should be treated initially with non-carbapenem-based antipseudomonal therapy taking into consideration antimicrobial stewardship. Non-azole antifungal drugs, including caspofungin and liposomal amphotericin B, should be preferred as empirical antifungal therapy in the events of possible or probable invasive fungal infections with an absence of pulmonary findings due to increase azole resistance. PMID:25258539

Synergistic antibacterial effect of silver and ebselen against multidrug-resistant Gram-negative bacterial infections.

PubMed

Zou, Lili; Lu, Jun; Wang, Jun; Ren, Xiaoyuan; Zhang, Lanlan; Gao, Yu; Rottenberg, Martin E; Holmgren, Arne

2017-08-01

Multidrug-resistant (MDR) Gram-negative bacteria account for a majority of fatal infections, and development of new antibiotic principles and drugs is therefore of outstanding importance. Here, we report that five most clinically difficult-to-treat MDR Gram-negative bacteria are highly sensitive to a synergistic combination of silver and ebselen. In contrast, silver has no synergistic toxicity with ebselen on mammalian cells. The silver and ebselen combination causes a rapid depletion of glutathione and inhibition of the thioredoxin system in bacteria. Silver ions were identified as strong inhibitors of Escherichia coli thioredoxin and thioredoxin reductase, which are required for ribonucleotide reductase and DNA synthesis and defense against oxidative stress. The bactericidal efficacy of silver and ebselen was further verified in the treatment of mild and acute MDR E. coli peritonitis in mice. These results demonstrate that thiol-dependent redox systems in bacteria can be targeted in the design of new antibacterial drugs. The silver and ebselen combination offers a proof of concept in targeting essential bacterial systems and might be developed for novel efficient treatments against MDR Gram-negative bacterial infections. © 2017 The Authors. Published under the terms of the CC BY 4.0 license.

Antibiotic-non-antibiotic combinations for combating extremely drug-resistant Gram-negative 'superbugs'.

PubMed

Schneider, Elena K; Reyes-Ortega, Felisa; Velkov, Tony; Li, Jian

2017-02-28

The emergence of antimicrobial resistance of Gram-negative pathogens has become a worldwide crisis. The status quo for combating resistance is to employ synergistic combinations of antibiotics. Faced with this fast-approaching post-antibiotic era, it is critical that we devise strategies to prolong and maximize the clinical efficacy of existing antibiotics. Unfortunately, reports of extremely drug-resistant (XDR) Gram-negative pathogens have become more common. Combining antibiotics such as polymyxin B or the broad-spectrum tetracycline and minocycline with various FDA-approved non-antibiotic drugs have emerged as a novel combination strategy against otherwise untreatable XDR pathogens. This review surveys the available literature on the potential benefits of employing antibiotic-non-antibiotic drug combination therapy. The apex of this review highlights the clinical utility of this novel therapeutic strategy for combating infections caused by 'superbugs'. © 2017 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Multidrug resistance in enteric and other gram-negative bacteria.

PubMed

George, A M

1996-05-15

In Gram-negative bacteria, multidrug resistance is a term that is used to describe mechanisms of resistance by chromosomal genes that are activated by induction or mutation caused by the stress of exposure to antibiotics in natural and clinical environments. Unlike plasmid-borne resistance genes, there is no alteration or degradation of drugs or need for genetic transfer. Exposure to a single drug leads to cross-resistance to many other structurally and functionally unrelated drugs. The only mechanism identified for multidrug resistance in bacteria is drug efflux by membrane transporters, even though many of these transporters remain to be identified. The enteric bacteria exhibit mostly complex multidrug resistance systems which are often regulated by operons or regulons. The purpose of this review is to survey molecular mechanisms of multidrug resistance in enteric and other Gram-negative bacteria, and to speculate on the origins and natural physiological functions of the genes involved.

Surface-Layer (S-Layer) Proteins Sap and EA1 Govern the Binding of the S-Layer-Associated Protein BslO at the Cell Septa of Bacillus anthracis

PubMed Central

Kern, Valerie J.; Kern, Justin W.; Theriot, Julie A.; Schneewind, Olaf

2012-01-01

The Gram-positive pathogen Bacillus anthracis contains 24 genes whose products harbor the structurally conserved surface-layer (S-layer) homology (SLH) domain. Proteins endowed with the SLH domain associate with the secondary cell wall polysaccharide (SCWP) following secretion. Two such proteins, Sap and EA1, have the unique ability to self-assemble into a paracrystalline layer on the surface of bacilli and form S layers. Other SLH domain proteins can also be found within the S layer and have been designated Bacillus S-layer-associated protein (BSLs). While both S-layer proteins and BSLs bind the same SCWP, their deposition on the cell surface is not random. For example, BslO is targeted to septal peptidoglycan zones, where it catalyzes the separation of daughter cells. Here we show that an insertional lesion in the sap structural gene results in elongated chains of bacilli, as observed with a bslO mutant. The chain length of the sap mutant can be reduced by the addition of purified BslO in the culture medium. This complementation in trans can be explained by an increased deposition of BslO onto the surface of sap mutant bacilli that extends beyond chain septa. Using fluorescence microscopy, we observed that the Sap S layer does not overlap the EA1 S layer and slowly yields to the EA1 S layer in a growth-phase-dependent manner. Although present all over bacilli, Sap S-layer patches are not observed at septa. Thus, we propose that the dynamic Sap/EA1 S-layer coverage of the envelope restricts the deposition of BslO to the SCWP at septal rings. PMID:22609927

[Etiology of pathogen-induced changes between the toes in patients working under moist conditions].

PubMed

Gründer, K; Kroker, A; Heppner, M; Marre, R

1988-11-15

Referring to recent studies, we discuss the epidemiology of tinea pedis and its position within the range of polyetiological foot infections caused by microbial agents. 201 industrial workers used to wear rubber boots because of moist working conditions were examined with regard to their feet. 107 of them showed clinically altered toe web; in 45 cases, we found ringworm infection (22.4%), in correlation to increased daily wearing time of rubber boots. 85 workers showed colonization of pathogenic bacteria (42.4%), especially gram-negative species (25.8%). Frequently, mixed infections were found. The clinical picture not always allows definite conclusions as to the causal agents. The symptom "maceration" is seen in each of the 3 pathogenic groups of germs (fungi, gram-positive pathogenic cocci, gram-negative bacteria), frequently in bacterial foot infections, especially gram-negative infections. Marked clinical symptoms involve increased infection rates. The predisposing factors to foot infection such as interdigital maceration and the promoting role of rubber boots have been confirmed. The final diagnosis is to be reassured by cultural investigations, in order to set up a specific treatment.

Nosocomial pneumonia in the ICU--year 2000 and beyond.

PubMed

Bowton, D L

1999-03-01

Diagnostic and treatment strategies in ICU patients with ventilator-associated pneumonia (VAP) remain controversial, largely because of the paucity of well-controlled comparison trials using clinically important end points. Recent studies indicating that early appropriate antibiotic therapy significantly lowers mortality underscore the urgent need for well-designed comparative trials. When quantitatively cultured, bronchial specimens obtained by noninvasive techniques may provide clinically useful information and avoid the higher costs and risks of invasive bronchoscopic diagnostic techniques. Previous antibiotic use before onset of nosocomial pneumonia raises the likelihood of infection with highly virulent organisms, such as Pseudomonas aeruginosa and Acinetobacter sp. Thus, the empiric antibiotic regimen should be active against these Gram-negative pathogens as well as other common Gram-negative and Gram-positive causative organisms. Promising preventive modalities for nosocomial VAP include use of a semirecumbent position, endotracheal tubes that allow continuous aspiration of secretions, and heat and moisture exchangers. Rotating their standard empiric antibiotic regimens and restricting the use of third-generation cephalosporins as empiric therapy may help hospitals reduce the incidence of nosocomial pneumonia caused by resistant Gram-negative pathogens.

Gram-negative bacteria that produce carbapenemases causing death attributed to recent foreign hospitalization.

PubMed

Ahmed-Bentley, Jasmine; Chandran, A Uma; Joffe, A Mark; French, Desiree; Peirano, Gisele; Pitout, Johann D D

2013-07-01

Overseas travel, as a risk factor for the acquisition of infections due to antimicrobial-resistant organisms, has recently been linked to carbapenemase-producing Gram-negative bacteria. Multiresistant Klebsiella pneumoniae, Escherichia coli, and Acinetobacter baumannii strains were isolated from a wound of a Canadian patient with a recent history of hospitalization in India. This resulted in the initiation of outbreak management that included surveillance cultures. Epidemiological and molecular investigations showed that NDM-1-producing K. pneumoniae ST16 and OXA-23-producing A. baumannii ST10 strains were transmitted to 5 other patients, resulting in the colonization of 4 patients and the death of 1 patient due to septic shock caused by the OXA-23-producing A. baumannii strain. The high rate of false positivity of the screening cultures resulted in additional workloads and increased costs for infection control and clinical laboratory work. We believe that this is the first report of an infection with carbapenemase-producing Gram-negative bacteria resulting in death attributed to a patient with recent foreign hospitalization. We recommend routine rectal and wound screening for colonization with multiresistant bacteria for patients who have recently been admitted to hospitals outside Canada.

Antibiofilm effect of Nocardiopsis sp. GRG 1 (KT235640) compound against biofilm forming Gram negative bacteria on UTIs.

PubMed

Rajivgandhi, Govindan; Vijayan, Ramachandran; Maruthupandy, Muthuchamy; Vaseeharan, Baskaralingam; Manoharan, Natesan

2018-05-01

Urinary tract infections (UTIs) are diverse public health complication and caused by range of pathogens, however mostly Gram negative bacteria cause significant life threatening risks to different populations. The prevalence rate and antimicrobial resistance among the Gram negative uropathogens alarmed significantly heighten the economic burden of these infections. In this study, we investigated the antibiofilm efficiency of Pyrrolo [1,2-a] pyrazine-1,4-dione,hexahydro-3-(2-methylpropyl) extracted from endophytic actinomycetes Nocardiopsis sp. GRG 1 (KT235640) against P. mirabilis and E. coli. The extracted compound was characterized through TLC, HPLC, GC-MS, LC-MS and confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM). The compound, Pyrrolo [1,2-a] pyrazine-1, 4-dione, hexahydro-3-(2-methylpropyl) inhibits both bacterial biofilm formation as well as reduces the viability of preformed biofilms. Furthermore, CLSM image shows cell shrinkage, disorganized cell membrane and loss of viability. The SEM result also confirms the cell wall degradation in treated cells of the bacteria. Hence, the Pyrrolo [1,2-a]pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl) is active against P. mirabilis and E. coli. Copyright © 2018 Elsevier Ltd. All rights reserved.

Bioenergetics and the role of soluble cytochromes C for alkaline adaptation in gram-negative alkaliphilic Pseudomonas.

PubMed

Matsuno, T; Yumoto, I

2015-01-01

Very few studies have been conducted on alkaline adaptation of Gram-negative alkaliphiles. The reversed difference of H(+) concentration across the membrane will make energy production considerably difficult for Gram-negative as well as Gram-positive bacteria. Cells of the alkaliphilic Gram-negative bacterium Pseudomonas alcaliphila AL15-21(T) grown at pH 10 under low-aeration intensity have a soluble cytochrome c content that is 3.6-fold higher than that of the cells grown at pH 7 under high-aeration intensity. Cytochrome c-552 content was higher (64% in all soluble cytochromes c) than those of cytochrome c-554 and cytochrome c-551. In the cytochrome c-552-dificient mutant grown at pH 10 under low-aeration intensity showed a marked decrease in μ max⁡ [h(-1)] (40%) and maximum cell turbidity (25%) relative to those of the wild type. Considering the high electron-retaining abilities of the three soluble cytochromes c, the deteriorations in the growth of the cytochrome c-552-deficient mutant could be caused by the soluble cytochromes c acting as electron storages in the periplasmic space of the bacterium. These electron-retaining cytochromes c may play a role as electron and H(+) condenser, which facilitate terminal oxidation at high pH under air-limited conditions, which is difficult to respire owing to less oxygen and less H(+).

Bioenergetics and the Role of Soluble Cytochromes c for Alkaline Adaptation in Gram-Negative Alkaliphilic Pseudomonas

PubMed Central

Matsuno, T.; Yumoto, I.

2015-01-01

Very few studies have been conducted on alkaline adaptation of Gram-negative alkaliphiles. The reversed difference of H+ concentration across the membrane will make energy production considerably difficult for Gram-negative as well as Gram-positive bacteria. Cells of the alkaliphilic Gram-negative bacterium Pseudomonas alcaliphila AL15-21T grown at pH 10 under low-aeration intensity have a soluble cytochrome c content that is 3.6-fold higher than that of the cells grown at pH 7 under high-aeration intensity. Cytochrome c-552 content was higher (64% in all soluble cytochromes c) than those of cytochrome c-554 and cytochrome c-551. In the cytochrome c-552-dificient mutant grown at pH 10 under low-aeration intensity showed a marked decrease in μ max⁡ [h−1] (40%) and maximum cell turbidity (25%) relative to those of the wild type. Considering the high electron-retaining abilities of the three soluble cytochromes c, the deteriorations in the growth of the cytochrome c-552-deficient mutant could be caused by the soluble cytochromes c acting as electron storages in the periplasmic space of the bacterium. These electron-retaining cytochromes c may play a role as electron and H+ condenser, which facilitate terminal oxidation at high pH under air-limited conditions, which is difficult to respire owing to less oxygen and less H+. PMID:25705691

Automatic detection of mycobacterium tuberculosis using artificial intelligence

PubMed Central

Xiong, Yan; Ba, Xiaojun; Hou, Ao; Zhang, Kaiwen; Chen, Longsen

2018-01-01

Background Tuberculosis (TB) is a global issue that seriously endangers public health. Pathology is one of the most important means for diagnosing TB in clinical practice. To confirm TB as the diagnosis, finding specially stained TB bacilli under a microscope is critical. Because of the very small size and number of bacilli, it is a time-consuming and strenuous work even for experienced pathologists, and this strenuosity often leads to low detection rate and false diagnoses. We investigated the clinical efficacy of an artificial intelligence (AI)-assisted detection method for acid-fast stained TB bacillus. Methods We built a convolutional neural networks (CNN) model, named tuberculosis AI (TB-AI), specifically to recognize TB bacillus. The training set contains 45 samples, including 30 positive cases and 15 negative cases, where bacilli are labeled by human pathologists. Upon training the neural network model, 201 samples (108 positive cases and 93 negative cases) were collected as test set and used to examine TB-AI. We compared the diagnosis of TB-AI to the ground truth result provided by human pathologists, analyzed inconsistencies between AI and human, and adjusted the protocol accordingly. Trained TB-AI were run on the test data twice. Results Examined against the double confirmed diagnosis by pathologists both via microscopes and digital slides, TB-AI achieved 97.94% sensitivity and 83.65% specificity. Conclusions TB-AI can be a promising support system to detect stained TB bacilli and help make clinical decisions. It holds the potential to relieve the heavy workload of pathologists and decrease chances of missed diagnosis. Samples labeled as positive by TB-AI must be confirmed by pathologists, and those labeled as negative should be reviewed to make sure that the digital slides are qualified. PMID:29707349

Automatic detection of mycobacterium tuberculosis using artificial intelligence.

PubMed

Xiong, Yan; Ba, Xiaojun; Hou, Ao; Zhang, Kaiwen; Chen, Longsen; Li, Ting

2018-03-01

Tuberculosis (TB) is a global issue that seriously endangers public health. Pathology is one of the most important means for diagnosing TB in clinical practice. To confirm TB as the diagnosis, finding specially stained TB bacilli under a microscope is critical. Because of the very small size and number of bacilli, it is a time-consuming and strenuous work even for experienced pathologists, and this strenuosity often leads to low detection rate and false diagnoses. We investigated the clinical efficacy of an artificial intelligence (AI)-assisted detection method for acid-fast stained TB bacillus. We built a convolutional neural networks (CNN) model, named tuberculosis AI (TB-AI), specifically to recognize TB bacillus. The training set contains 45 samples, including 30 positive cases and 15 negative cases, where bacilli are labeled by human pathologists. Upon training the neural network model, 201 samples (108 positive cases and 93 negative cases) were collected as test set and used to examine TB-AI. We compared the diagnosis of TB-AI to the ground truth result provided by human pathologists, analyzed inconsistencies between AI and human, and adjusted the protocol accordingly. Trained TB-AI were run on the test data twice. Examined against the double confirmed diagnosis by pathologists both via microscopes and digital slides, TB-AI achieved 97.94% sensitivity and 83.65% specificity. TB-AI can be a promising support system to detect stained TB bacilli and help make clinical decisions. It holds the potential to relieve the heavy workload of pathologists and decrease chances of missed diagnosis. Samples labeled as positive by TB-AI must be confirmed by pathologists, and those labeled as negative should be reviewed to make sure that the digital slides are qualified.

Association of high mortality with extended-spectrum β-lactamase (ESBL) positive cultures in community acquired infections.

PubMed

Ray, Sumit; Anand, Dimple; Purwar, Sankalp; Samanta, Arijit; Upadhye, Kaustubh V; Gupta, Prasoon; Dhar, Debashis

2018-04-01

Infections due to multidrug resistant organisms have become a serious health concern worldwide. The present study was conducted to investigate the spectrum of microbial resistance pattern in the community and their effects on mortality. A retrospective review and analysis of prospectively collected data was done of all patients admitted with diagnosis of sepsis in two tertiary care ICU's for a period of two years. Demographics, culture positivity, microbial spectrum, resistance pattern and outcome data were collected. Out of 5309 patients enrolled; 3822 had suspected clinical infection on admission with 1452 patients growing positive microbial cultures. Among these, 201 bacterial strains were isolated from patients who had community acquired infections. 73% were Gram negative bacilli, commonest being E. coli (63%). 63.4% E. coli and 60.7% Klebsiella isolates were ESBL producers. The mortality in ESBL positive infections was significantly higher as compared to ESBL negative infections (Odds ratio 2.756). Moreover, ESBL positive patients empirically treated with Beta Lactams+Beta Lactamase inhibitors (BL+BLI) had significantly higher mortality as compared to patients treated with carbapenems. More data from multiple centres need to be gathered to formulate appropriate antibiotic policy for critically ill patients admitted from the community. Copyright © 2017 Elsevier Inc. All rights reserved.

Microbiological Aetiology, Epidemiology, and Clinical Profile of Prosthetic Joint Infections: Are Current Antibiotic Prophylaxis Guidelines Effective?

PubMed Central

Cheng, Allen C.; Buising, Kirsty L.; Choong, Peter F. M.

2012-01-01

Prosthetic joint infections remain a major complication of arthroplasty. At present, local and international guidelines recommend cefazolin as a surgical antibiotic prophylaxis at the time of arthroplasty. This retrospective cohort study conducted across 10 hospitals over a 3-year period (January 2006 to December 2008) investigated the epidemiology and microbiological etiology of prosthetic joint infections. There were 163 cases of prosthetic joint infection identified. From a review of the microbiological culture results, methicillin-resistant Staphylococcus aureus (MRSA) and coagulase-negative staphylococci were isolated in 45% of infections. In addition, polymicrobial infections, particularly those involving Gram-negative bacilli and enterococcal species, were common (36%). The majority (88%) of patients received cefazolin as an antibiotic prophylaxis at the time of arthroplasty. In 63% of patients in this cohort, the microorganisms subsequently obtained were not susceptible to the antibiotic prophylaxis administered. The results of this study highlight the importance of ongoing reviews of the local ecology of prosthetic joint infection, demonstrating that the spectrum of pathogens involved is broad. The results should inform empirical antibiotic therapy. This report also provokes discussion about infection control strategies, including changing surgical antibiotic prophylaxis to a combination of glycopeptide and cefazolin, to reduce the incidence of infections due to methicillin-resistant staphylococci. PMID:22314530

Use of Positive Blood Cultures for Direct Identification and Susceptibility Testing with the Vitek 2 System

PubMed Central

de Cueto, Marina; Ceballos, Esther; Martinez-Martinez, Luis; Perea, Evelio J.; Pascual, Alvaro

2004-01-01

In order to further decrease the time lapse between initial inoculation of blood culture media and the reporting of results of identification and antimicrobial susceptibility tests for microorganisms causing bacteremia, we performed a prospective study in which specially processed fluid from positive blood culture bottles from Bactec 9240 (Becton Dickinson, Cockeysville, Md.) containing aerobic media were directly inoculated into Vitek 2 system cards (bio-Mérieux, France). Organism identification and susceptibility results were compared with those obtained from cards inoculated with a standardized bacterial suspension obtained following subculture to agar; 100 consecutive positive monomicrobic blood cultures, consisting of 50 gram-negative rods and 50 gram-positive cocci, were included in the study. For gram-negative organisms, 31 of the 50 (62%) showed complete agreement with the standard method for species identification, while none of the 50 gram-positive cocci were correctly identified by the direct method. For gram-negative rods, there were 50% categorical agreements between the direct and standard methods for all drugs tested. The very major error rate was 2.4%, and the major error rate was 0.6%. The overall error rate for gram-negatives was 6.6%. Complete agreement in clinical categories of all antimicrobial agents evaluated was obtained for 19 of 50 (38%) gram-positive cocci evaluated; the overall error rate was 8.4%, with 2.8% minor errors, 2.4% major errors, and 3.2% very major errors. These findings suggest that the Vitek 2 cards inoculated directly from positive Bactec 9240 bottles do not provide acceptable bacterial identification or susceptibility testing in comparison with corresponding cards tested by a standard method. PMID:15297523

Intestinal volvulus with coagulative hepatic necrosis in a chicken.

PubMed

Haridy, Mohie; Goryo, Masanobu; Sasaki, Jun; Okada, Kosuke

2010-04-01

A 7-week-old SPF chicken inoculated at 4 weeks of age with chicken anemia virus was puffed up depressed and had ruffled feathers and a good body condition. Intestinal volvulus involving the jejunum and part of the duodenum forming two loops with one knob was observed. Microscopically, venous infarction of the obstructed loops, periportal and sublobular multifocal coagulative hepatic necrosis and granulomatous inflammation of the cecal tonsils were observed. Gram staining revealed no bacteria in hepatic tissue; however, gram-positive bacilli were detected in the necrotic debris in the intestinal lumen. Immunosuppression might have predisposed the chicken to intestinal and cecal tonsil infection that then progressed to volvulus. Loss of the mucosal barrier in infarction might allow bacterial toxins and vasoactive factors to escape into the systemic circulation (toxemia) and be responsible for the hepatic necrosis.

[Non-species-specific effects of unacylated homoserine lactone and hexylresorcinol, low molecular weight autoregulators, on the growth and development of bacteria].

PubMed

Muliukin, A L; Filippova, S N; Kozlova, A N; Surgucheva, N A; Bogdanova, T I; Tsaplina, I A; El'-Registan, G I

2006-01-01

We conducted a comparative study of the effects of alpha-amino-gamma-butyrolactone, the common structural element of extracellular microbial regulators of the homoserine lactone (HSL) group, and of 4-n-hexylresorcinol, an autoregulator of the alkylhydroxybenzene (AHB) group, on the growth and development of gram-positive and gram-negative bacteria. We revealed non-species-specific effects of HSL and AHB and characterized their concentration dependencies. The addition of 10(-5)-10(-3) M HSL or 10(-5)-10(-4) M AHB during the exponential growth phase of the cultures grown on balanced media resulted in cell division arrest and accelerated the transition to the stationary phase that culminated in endospore formation in Bacillus cereus, Alicyclobacillus tolerans, and Sulfobacillus thermosulfidooxidans. When bacilli grew under the cultivation conditions that resulted in a low-zero spore percentage, 10(-4)-10(-3) M HSL cancelled the inhibition of spore formation. In the gram-negative bacteria Pseudomonas aurantiaca and Azotobacter vinelandii, AHB at concentrations of 10(-4) to (1.5-2.5) 10(-4) M induced the formation of dormant cells. Studies with the actinobacterium Streptomyces avermitilis revealed that the HSL effect varied depending on the age of the test cultures. The addition of 10(-4) M HSL during the lag phase of a submerged streptomycete culture accelerated its transition to the stationary phase and induced the formation of endospores, the dormant cells that are regarded as alternatives to exospores (conidia). If HSL (3.64 and 4.55 mg per 1cm2 disc) was locally added to a surface S. avermitilis culture, the growing mycelium formed rings that differed in their density, in the extent of the development of aerial mycelium, and in the presence/absence of exospores. Ring-shaped growth of streptomycete mycelia was also induced by 0.075-0.75 mg of AHB; however, unlike HSL, AHB repressed exospore formation. The data on non-species-specific effects of HSL and AHB suggest that they may perform regulatory functions on the microbial community level.

Carbapenem stewardship: positive impact on hospital ecology.

PubMed

Lima, Ana Lucia Lei Munhoz; Oliveira, Priscila Rosalba Domingos de; Paula, Adriana Pereira de; Dal-Paz, Karine; Almeida, João Nóbrega de; Félix, Cássia da Silva; Rossi, Flávia

2011-01-01

Excessive group 2 carbapenem use may result in decreased bacterial susceptibility. We evaluated the impact of a carbapenem stewardship program, restricting imipenem and meropenem use. Ertapenem was mandated for ESBL-producing Enterobacteriaceae infections in the absence of non-fermenting Gram-negative bacilli (GNB) from April 2006 to March 2008. Group 2 carbapenems were restricted for use against GNB infections susceptible only to carbapenems and suspected GNB infections in unstable patients. Cumulative susceptibility tests were done for nosocomial pathogens before and after restriction using Clinical and Laboratory Standards Institute (CLSI) guide-lines.Vitek System or conventional identification methods were performed and susceptibility testing done by disk diffusion according to CLSI.Antibiotic consumption (t-test) and susceptibilities (McNemar's test) were determined. The defined daily doses (DDD) of group 2 carbapenems declined from 61.1 to 48.7 DDD/1,000 patient-days two years after ertapenem introduction (p = 0.027). Mean ertapenem consumption after restriction was 31.5 DDD/1,000 patient-days. Following ertapenem introduction no significant susceptibility changes were noticed among Gram-positive cocci. The most prevalent GNB were P. aeruginosa, Klebsiella pneumoniae, and Acinetobacter spp. There was no change in P. aeruginosa susceptibility to carbapenems. Significantly improved P. aeruginosa and K. pneumoniae ciprofloxacin susceptibilities were observed, perhaps due to decreased group 2 carbapenem use. K. pneumoniae susceptibility to trimethoprim-sulfamethoxazole improved. Preferential use of ertapenem resulted in reduced group 2 carbapenem use, with a positive impact on P. aeruginosa and K. pneumoniae susceptibility.

Bacterial profile in primary teeth with necrotic pulp and periapical lesions.

PubMed

da Silva, Léa Assed Bezerra; Nelson-Filho, Paulo; Faria, Gisele; de Souza-Gugelmin, Maria Cristina Monteiro; Ito, Izabel Yoko

2006-01-01

The objective of this study was to evaluate the bacterial profile in root canals of human primary teeth with necrotic pulp and periapical lesions using bacterial culture. A total of 20 primary teeth with necrotic pulp and radiographically visible radiolucent areas in the region of the bone furcation and/or the periapical region were selected. After crown access, 4 sterile absorbent paper points were introduced sequentially into the root canal for collection of material. After 30 s, the paper points were removed and placed in a test tube containing reduced transport fluid (RTF) and were sent for microbiological evaluation. Anaerobic microorganisms were found in 100% of the samples, black-pigmented bacilli in 30%, aerobic microorganisms in 60%, streptococci in 85%, gram-negative aerobic rods in 15% and staphylococci were not quantified. Mutans streptococci were found in 6 root canals (30%), 5 canals with Streptococcus mutans and 1 canal with Streptococcus mutans and Streptococcus sobrinus. It was concluded that in root canals of human primary teeth with necrotic pulp and periapical lesions, the infection is polymicrobial with predominance of anaerobic microorganisms.

[Complicated urinary tract infections--from the perspective of the medical technologist].

PubMed

Nagasawa, Zenzo

2002-07-01

We would like to propose re-establishment of the protocol for ordering a clinical microbiology laboratory test after a bedside screening test using urine reagent strip when urinary tract infection is suspected. Media for isolation shall be chosen by the clinical microbiology laboratory after checking turbidity and microscopic examination of the urine specimen. In cases of complicated urinary tract infections, quantitative culture should be performed to investigate changes in the number of microorganism to grasp condition of super infection. In such infections, there are many cases in which multiple microorganism growth including glucose non-fermenting gram-negative bacilli can be recognized. Therefore, it is necessary to inspect colonies on media as long as possible (24 hrs culture may be short in some cases). The protocol for microorganism identification and susceptibility test for such specimen varies in each laboratory, considering the Health Insurance Point System (reimbursement system by MHW). It is necessary to communicate with physicians and to refer to past results to proceed with the laboratory test properly. Therefore, a Certified Clinical Microbiology Medical Technologist is needed and the role played by such staff is important.

The importance of soap selection for routine hand hygiene in hospital.

PubMed Central

Ojajärvi, J.

1981-01-01

Five different types of liquid soap were studied in hospital wards, each during two months' use. Altogether 1306 finger print samples were taken from the hands of the staff by sampling twice a week and the acceptability of the soaps was measured by a questionnaire. During the use of different soaps only slight differences were found in the numbers of total bacteria or in the occurrence of Staph. aureus and gram-negative bacilli on the hands. During the use of the emulsion-type product studied, several persons who had dermatological problems had lower mean bacterial counts of the fingers than during the use of the other soaps. This soap was also favourably accepted by the staff. After over one year's use of pine oil soap and alcohol, the staff of the hospital was satisfied with the method. However, several persons with skin problems admitted to not using soap or alcohol. The considerable differences found in the acceptability of soaps imply that for use in hospital the choice of a soap acceptable to the nursing staff is important in promoting proper hand hygiene. PMID:7240731

The effect of a changed environment on bacterial colonization rates in an established burns centre

PubMed Central

Wormald, P. J.

1970-01-01

In an established burns centre which moved from an old building to new purpose-designed premises, colonization rates of patients' burns with Staphylococcus aureus, Pseudomonas aeruginosa and other Gram-negative bacilli were not reduced. Colonization rates with Streptococcus pyogenes increased but the increase was mainly due to multiple importations in the new premises of a strain of higher communicability than any seen in the old. In the first 32 months in the new environment 10 patients were found colonized with pseudomonas on admission and 20 became colonized in the unit. A much higher proportion of patients with burns of more than 30% body surface became colonized than of patients with less. About one-third of the above 20 patients became colonized with strains already isolated from another patient; all but one of them had small area burns. Cross-infection was not observed from numerous heavily colonized patients with high percentage burns. This paradox is discussed in detail. Basin outflows in the new premises became colonized with P. aeruginosa of two serotypes not found on patients in this unit. PMID:4993291

The effect of a changed environment on bacterial colonization rates in an established burns centre.

PubMed

Wormald, P J

1970-12-01

In an established burns centre which moved from an old building to new purpose-designed premises, colonization rates of patients' burns with Staphylococcus aureus, Pseudomonas aeruginosa and other Gram-negative bacilli were not reduced. Colonization rates with Streptococcus pyogenes increased but the increase was mainly due to multiple importations in the new premises of a strain of higher communicability than any seen in the old.In the first 32 months in the new environment 10 patients were found colonized with pseudomonas on admission and 20 became colonized in the unit. A much higher proportion of patients with burns of more than 30% body surface became colonized than of patients with less. About one-third of the above 20 patients became colonized with strains already isolated from another patient; all but one of them had small area burns. Cross-infection was not observed from numerous heavily colonized patients with high percentage burns. This paradox is discussed in detail. Basin outflows in the new premises became colonized with P. aeruginosa of two serotypes not found on patients in this unit.

Infectious crystalline keratopathy associated with Klebsiella oxytoca.

PubMed

Chou, Timothy Y; Adyanthaya, Rohit

2012-12-01

The purpose of this study is to report a novel case of a Klebsiella oxytoca-associated infectious crystalline keratopathy This is a case report study. An 80-year-old woman presented with complaint of noticing a white spot in the left eye for 2 to 3 days, as well as mild soreness and discharge. Past ocular history was notable for a failed left corneal transplant for which she was taking prednisolone acetate 1 % twice per day. On slit-lamp examination, there was an extensive stromal ulcer and infiltrate in the inferior half of the transplant. Extending superiorly in the graft were branching, needle-like deep stromal opacities, characteristic of infectious crystalline keratopathy. Diagnostic scrapings revealed Gram-negative bacilli, subsequently identified on culture as K. oxytoca. There was also light growth of Staphylococcus species. The patient was placed on double topical antibiotic therapy with moxifloxacin and fortified tobramycin. After 2 months of treatment there was gradual resolution of the infection. K. oxytoca is a microorganism which can be associated with clinical infectious crystalline keratopathy, presenting as a mixed infection along with Staphylococcus species.

Gram staining for the treatment of peritonsillar abscess.

PubMed

Takenaka, Yukinori; Takeda, Kazuya; Yoshii, Tadashi; Hashimoto, Michiko; Inohara, Hidenori

2012-01-01

Objective. To examine whether Gram staining can influence the choice of antibiotic for the treatment of peritonsillar abscess. Methods. Between 2005 and 2009, a total of 57 cases of peritonsillar abscess were analyzed with regard to cultured bacteria and Gram staining. Results. Only aerobes were cultured in 16% of cases, and only anaerobes were cultured in 51% of cases. Mixed growth of aerobes and anaerobes was observed in 21% of cases. The cultured bacteria were mainly aerobic Streptococcus, anaerobic Gram-positive cocci, and anaerobic Gram-negative rods. Phagocytosis of bacteria on Gram staining was observed in 9 cases. The bacteria cultured from these cases were aerobic Streptococcus, anaerobic Gram-positive cocci, and anaerobic Gram-negative rods. The sensitivity of Gram staining for the Gram-positive cocci and Gram-negative rods was 90% and 64%, respectively. The specificity of Gram staining for the Gram-positive cocci and Gram-negative rods was 62% and 76%, respectively. Most of the Gram-positive cocci were sensitive to penicillin, but some of anaerobic Gram-negative rods were resistant to penicillin. Conclusion. When Gram staining shows only Gram-positive cocci, penicillin is the treatment of choice. In other cases, antibiotics effective for the penicillin-resistant organisms should be used.

Emphysematous endocarditis caused by AmpC beta-lactamase-producing Escherichia coli: A case report.

PubMed

Kim, Chung-Jong; Yi, Jeong-Eun; Kim, Yookyung; Choi, Hee Jung

2018-02-01

Infective endocarditis (IE) is a life-threatening disease, mostly caused by gram-positive bacteria. Gram-negative bacteria were identified as a causative organism in relatively small number of cases. Although, antibiotic-resistant Escherichia coli is common cause of gram-negative endocarditis, AmpC beta-lactamase (BL)-harboring E coli is very rare cause of IE. Furthermore, emphysematous endocarditis is also a very rare manifestation of E coli infection. We report a case of 80-year-old female patient presenting with dizziness, fever, and altered mental status, who was finally diagnosed with emphysematous endocarditis caused by E coli harboring an AmpC BL gene. Her chest computed tomography revealed air bubbles surrounding the annulus of a mitral valve and a transesophageal echocardiogram revealed a hyperechogenic mass fixed on the posteromedial side of the mitral annulus with 2 eccentric mitral regurgitation jets. Blood cultures grew E coli which harbored the DHA-type AmpC BL. The organism belonged to a B2 phylogenic group, and multilocus sequence typing analyses revealed that the strains were of ST-95. She was treated with meropenem following the resistant profiles, and surgery was recommended by the healthcare professional, but denied by the patient's guardians. She was transferred to another hospital due to a refusal for further treatment. Emphysematous endocarditis is an uncommon complication of E coli bacteremia. Certain phylogenetic groups may be associated with development of E coli endocarditis.

Clostridium perfringens type A fatal acute hemorrhagic gastroenteritis in a dog

PubMed Central

Schlegel, Ben J.; Van Dreumel, Tony; Slavić, Durda; Prescott, John F.

2012-01-01

The morning after participating in a dog show, a 2-year-old Pomeranian dog was found dead in a pool of bloody feces. Necropsy revealed hemorrhagic gastroenteritis of the entire gastrointestinal tract, with many Gram-positive bacilli on the surface and in the lumen and crypts of the intestine. Enterotoxin-positive type A Clostridium perfringens were isolated in large numbers. This dramatic case of fatal C. perfringens gastroenteritis highlights the need to better understand the role of this bacterium in enteric disease of dogs. PMID:23115371

Imaging Enterobacteriaceae infection in vivo with 18F-fluorodeoxysorbitol positron emission tomography

PubMed Central

Weinstein, Edward A.; Ordonez, Alvaro A.; DeMarco, Vincent P.; Murawski, Allison M.; Pokkali, Supriya; MacDonald, Elizabeth M.; Klunk, Mariah; Mease, Ronnie C.; Pomper, Martin G.; Jain, Sanjay K.

2015-01-01

The Enterobacteriaceae are a family of rod-shaped Gram-negative bacteria that normally inhabit the gastrointestinal tract and are the most common cause of Gram-negative bacterial infections in humans. In addition to causing serious multidrug-resistant, hospital-acquired infections, a number of Enterobacteriaceae species are also recognized as biothreat pathogens. As a consequence, new tools are urgently needed to specifically identify and localize infections due to Enterobacteriaceae and to monitor antimicrobial efficacy. In this report, we used commercially available 2-[18F]-fluorodeoxyglucose (18F-FDG) to produce 2-[18F]-fluorodeoxysorbitol (18F-FDS), a radioactive probe for Enterobacteriaceae, in 30 min. 18F-FDS selectively accumulated in Enterobacteriaceae, but not in Gram-positive bacteria or healthy mammalian or cancer cells in vitro. In a murine myositis model, 18F-FDS positron emission tomography (PET) rapidly differentiated true infection from sterile inflammation with a limit of detection of 6.2 ± 0.2 log10 colony-forming units (CFU) for Escherichia coli. Our findings were extended to models of mixed Gram-positive and Gram-negative thigh co-infections, brain infection, Klebsiella pneumonia, and mice undergoing immunosuppressive chemotherapy. This technique rapidly and specifically localized infections due to Enterobacteriaceae, providing a three-dimensional holistic view within the animal. Last, 18F-FDS PET monitored the efficacy of antimicrobial treatment, demonstrating a PET signal proportionate to the bacterial burden. Therapeutic failures associated with multidrug-resistant, extended-spectrum β-lactamase (ESBL)–producing E. coli infections were detected in real time. Together, these data show that 18F-FDS is a candidate imaging probe for translation to human clinical cases of known or suspected infections owing to Enterobacteriaceae. PMID:25338757

Factors influencing antimicrobial resistance and outcome of Gram-negative bloodstream infections in children.

PubMed

Ivády, Balázs; Kenesei, Éva; Tóth-Heyn, Péter; Kertész, Gabriella; Tárkányi, Klára; Kassa, Csaba; Ujhelyi, Enikő; Mikos, Borbála; Sápi, Erzsébet; Varga-Heier, Krisztina; Guóth, Gábor; Szabó, Dóra

2016-06-01

The aim of this study was to collect data about pediatric Gram-negative bloodstream infections (BSI) to determine the factors that influence multidrug resistance (MDR), clinical course and outcome of children affected by Gram-negative sepsis. In this observational, prospective, multicenter study we collected cases of pediatric Gram-negative BSI during a 2-year period. We analyzed epidemiological, microbiological and clinical factors that associated with acquisition of MDR infections and outcome. One-hundred and thirty-five BSI episodes were analyzed. Median age of children was 0.5 years (IQR 0.1-6.17, range 0-17 years). Predominant bacteria were Enterobacteriaceae (68.3 %), and Pseudomonas spp. (17.9 %). Multidrug resistance was detected in 45/134 cases (33.6 %), with the highest rates in Escherichia coli, Enterobacter and Pseudomonas spp. Acquisition of MDR pathogens was significantly associated with prior cephalosporin treatment, older age, admission to hemato-oncology unit, polymicrobial infections, higher rate of development of septic shock, and multiple organ failures. All-cause mortality was 17.9 %. Presence of septic shock at presentation and parenteral nutrition were associated with higher mortality. Pseudomonas spp., and Enterobacter spp. BSIs had the highest rate of mortality. Inappropriate empiric antibiotic therapy was more frequent in MDR patients, although not significantly associated with poor outcome. Rates of multidrug resistance and mortality in children with Gram-negative bloodstream infections remain high in our settings. Empiric broad-spectrum antibiotics and combination therapy could be recommended, especially in children with malignant diseases, patients admitted to the PICU, and for cases with septic shock, who have higher mortality risk.

Comparative activity of ceftobiprole against Gram-positive and Gram-negative isolates from Europe and the Middle East: the CLASS study.

PubMed

Rossolini, Gian M; Dryden, Matthew S; Kozlov, Roman S; Quintana, Alvaro; Flamm, Robert K; Läuffer, Jörg M; Lee, Emma; Morrissey, Ian; CLASS Study Group

2011-01-01

to assess the in vitro activity of ceftobiprole and comparators against a recent collection of Gram-positive and Gram-negative pathogens, in order to detect potential changes in susceptibility patterns, and to evaluate the Etest assay for ceftobiprole susceptibility testing. contemporary Gram-positive and Gram-negative isolates (excluding extended-spectrum β-lactamase-producing isolates) from across Europe and the Middle East were collected, and their susceptibility to ceftobiprole, vancomycin, teicoplanin, linezolid, ceftazidime and cefepime was assessed using the Etest method. Quality testing [using Etest and broth microdilution (BMD)] was conducted at a central reference laboratory. some 5041 Gram-positive and 4026 Gram-negative isolates were included. Against Gram-positive isolates overall, ceftobiprole had the lowest MIC50 (0.5 mg/L), compared with 1 mg/L for its comparators (vancomycin, teicoplanin and linezolid). Against methicillin-resistant Staphylococcus aureus, all four agents had a similar MIC90 (2 mg/L), but ceftobiprole had a 4-fold better MIC90 (0.5 mg/L) against methicillin-susceptible strains. Only 38 Gram-positive isolates were confirmed as ceftobiprole resistant. Among Gram-negative strains, 86.9%, 91.7% and 95.2% were susceptible to ceftobiprole, ceftazidime and cefepime, respectively. Pseudomonas aeruginosa was less susceptible to all three antimicrobials than any other Gram-negative pathogen. There was generally good agreement between local Etest results and those obtained at the reference laboratory (for ceftobiprole: 86.8% with Gram-negatives; and 94.7% with Gram-positives), as well as between results obtained by BMD and Etest methods (for ceftobiprole: 98.2% with Gram-negatives; and 98.4% with Gram-positives). ceftobiprole exhibits in vitro activity against a wide range of Gram-positive and Gram-negative pathogens, including multidrug-resistant strains. No changes in its known susceptibility profile were identified.

In vitro activity of tigecycline and comparators against a European collection of anaerobes collected as part of the Tigecycline Evaluation and Surveillance Trial (T.E.S.T.) 2010-2016.

PubMed

Rodloff, Arne C; Dowzicky, Michael J

2018-04-19

The Tigecycline Evaluation and Surveillance Trial (T.E.S.T.) is a global program that aims to monitor the in vitro antimicrobial activities of current therapeutic agents against clinical isolates. This study presents surveillance data for Gram-positive and Gram-negative anaerobic isolates (N = 7008) collected from nine European countries between 2010 and 2016. Presented in this study are antimicrobial susceptibility data, according to the European Committee for Antimicrobial Susceptibility Testing (EUCAST) breakpoints, and minimum inhibitory concentration (MIC) distributions. The antimicrobial agents tested were cefoxitin (Gram-negative isolates only), clindamycin, meropenem, metronidazole, penicillin (Gram-positive isolates only), piperacillin-tazobactam and tigecycline. Among all Gram-positive and Gram-negative anaerobes, the lowest rates of resistance were to meropenem and metronidazole (0.0%-1.7% and 0.0%-1.9%, respectively). High rates of resistance were reported to clindamycin, in particular among isolates of the Bacteroides fragilis group (22.1%-48.1%) and Prevotella spp. (10.9%-32.2%). The majority of MIC distributions were unimodal, with the exception of clindamycin, which were mostly bimodal. Fifty percent of Gram-negative isolates gave tigecycline MICs between 0.06 and 1 mg/L, and 50% of Gram-positive isolates exhibited tigecycline MICs between 0.06 and 0.25 mg/L. The findings of this study suggest that the majority of anaerobic isolates were susceptible to meropenem and metronidazole, and that tigecycline remained active, but clindamycin resistance is a cause for concern in Europe. Surveillance studies, such as T.E.S.T., provide information on changes in the susceptibility of clinically important pathogens to commonly prescribed antimicrobial agents, and can highlight problems of antimicrobial resistance that need to be addressed. Copyright © 2018 Pfizer Inc. Published by Elsevier Ltd.. All rights reserved.

TRIF Licenses Caspase-11-Dependent NLRP3 Inflammasome Activation by Gram-Negative Bacteria

PubMed Central

Rathinam, Vijay A.K.; Vanaja, Sivapriya Kailasan; Waggoner, Lisa; Sokolovska, Anna; Becker, Christine; Stuart, Lynda M.; Leong, John M.; Fitzgerald, Katherine A.

2013-01-01

SUMMARY Systemic infections with Gram-negative bacteria are characterized by high mortality rates due to the “sepsis syndrome,” a widespread and uncontrolled inflammatory response. Though it is well recognized that the immune response during Gram-negative bacterial infection is initiated after the recognition of endotoxin by Toll-like receptor 4, the molecular mechanisms underlying the detrimental inflammatory response during Gram-negative bacteremia remain poorly defined. Here, we identify a TRIF pathway that licenses NLRP3 inflammasome activation by all Gram-negative bacteria. By engaging TRIF, Gram-negative bacteria activate caspase-11. TRIF activates caspase-11 via type I IFN signaling, an event that is both necessary and sufficient for caspase-11 induction and autoactivation. Caspase-11 subsequently synergizes with the assembled NLRP3 inflammasome to regulate caspase-1 activation and leads to caspase-1-independent cell death. These events occur specifically during infection with Gram-negative, but not Gram-positive, bacteria. The identification of TRIF as a regulator of caspase-11 underscores the importance of TLRs as master regulators of inflammasomes during Gram-negative bacterial infection. PMID:22819539

Presence and destruction of tubercle bacilli in sewage*

PubMed Central

Jensen, K. Erik

1954-01-01

The author examined the sewage from 5 towns with tuberculosis sanatoria and from one institution for the care of the feeble-minded, which had a tuberculosis ward, for the presence of tubercle bacilli. The 6 effluents were treated in biological-purification plants and average samples taken. These were centrifuged, and the sediment treated for 1 hour at 37°C with 4% NaOH before inoculation into guinea-pigs. Tubercle bacilli were demonstrated in the influent to all the plants and in the digested sludge of all those operating on sewage where the ratio of infective patients to all persons connected with the plant was up to 1:600. Experiments with cultivated tubercle bacilli showed that centrifuging of sewage resulted in only an insignificant loss of bacilli, but that NaOH treatment caused a loss of over 99%. After consideration of the risk of infection to both man and cattle from the sewage of tuberculosis institutions, the author reports on his own studies on the killing of tubercle bacilli in sewage. It took about 11½-15 months before tubercle bacilli could no longer be demonstrated in sludge that had been kept on the drying beds. The addition of 10 mg of chlorine per litre of biologically purified effluent from an activated-sludge plant was found effectively to destroy tubercle bacilli. Disinfection of sludge was also carried out with 0.5% lysol and 0.1%-0.2% formol; 3.1% copper sulfate proved ineffective. The author concludes that the disinfection of sewage from tuberculosis institutions presents no special difficulties, but that work on this subject in different countries should be co-ordinated in an effort to improve plant and reduce costs. PMID:13160757

Examination of key intermediates in the catalytic cycle of aspartate-beta-semialdehyde dehydrogenase from a gram-positive infectious bacteria.

PubMed

Faehnle, Christopher R; Le Coq, Johanne; Liu, Xuying; Viola, Ronald E

2006-10-13

Aspartate-beta-semialdehyde dehydrogenase (ASADH) catalyzes a critical branch point transformation in amino acid bio-synthesis. The products of the aspartate pathway are essential in microorganisms, and this entire pathway is absent in mammals, making this enzyme an attractive target for antibiotic development. The first structure of an ASADH from a Gram-positive bacterium, Streptococcus pneumoniae, has now been determined. The overall structure of the apoenzyme has a similar fold to those of the Gram-negative and archaeal ASADHs but contains some interesting structural variations that can be exploited for inhibitor design. Binding of the coenzyme NADP, as well as a truncated nucleotide analogue, into an alternative conformation from that observed in Gram-negative ASADHs causes an enzyme domain closure that precedes catalysis. The covalent acyl-enzyme intermediate was trapped by soaking the substrate into crystals of the coenzyme complex, and the structure of this elusive intermediate provides detailed insights into the catalytic mechanism.

Polymicrobial and microsporidial keratitis in a patient using Boston scleral contact lens for Sjogren's syndrome and ocular cicatricial pemphigoid.

PubMed

Fernandes, Merle; Sharma, Savitri

2013-04-01

To report a rare case of microsporidial and polymicrobial keratitis in a patient with Sjogren's syndrome and ocular cicatricial pemphigoid. This is a descriptive case report. A 66-year-old lady diagnosed with Sjogren's syndrome (SS) and ocular cicatricial pemphigoid (OCP) presented to us with microbial keratitis after using a Boston sclera contact lens for a painful epithelial defect. After 9 days of medical treatment, she underwent therapeutic penetrating keratoplasty. 10% potassium hydroxide and calcofluor white wet mount revealed microsporidial spores. Gram positive cocci and Gram variable bacilli on Gram stain were identified as Staphylococcus epidermidis and Corynebacterium accolens in culture. Histopathological examination of the corneal tissue confirmed the presence of microsporidial spores. Microsporidal keratitis can occur in patients with severe ocular surface disease due to SS and OCP. Predisposing factors include dry eye, local and systemic immunosuppression and Boston scleral contact lens. Early surgical intervention may be needed to eradicate the infection. Copyright © 2012 British Contact Lens Association. Published by Elsevier Ltd. All rights reserved.

A New Class of Quorum Quenching Molecules from Staphylococcus Species Affects Communication and Growth of Gram-Negative Bacteria

PubMed Central

Chu, Ya-Yun; Nega, Mulugeta; Wölfle, Martina; Plener, Laure; Grond, Stephanie; Jung, Kirsten; Götz, Friedrich

2013-01-01

The knowledge that many pathogens rely on cell-to-cell communication mechanisms known as quorum sensing, opens a new disease control strategy: quorum quenching. Here we report on one of the rare examples where Gram-positive bacteria, the ‘Staphylococcus intermedius group’ of zoonotic pathogens, excrete two compounds in millimolar concentrations that suppress the quorum sensing signaling and inhibit the growth of a broad spectrum of Gram-negative beta- and gamma-proteobacteria. These compounds were isolated from Staphylococcus delphini. They represent a new class of quorum quenchers with the chemical formula N-[2-(1H-indol-3-yl)ethyl]-urea and N-(2-phenethyl)-urea, which we named yayurea A and B, respectively. In vitro studies with the N-acyl homoserine lactone (AHL) responding receptor LuxN of V. harveyi indicated that both compounds caused opposite effects on phosphorylation to those caused by AHL. This explains the quorum quenching activity. Staphylococcal strains producing yayurea A and B clearly benefit from an increased competitiveness in a mixed community. PMID:24098134

Brevundimonas spp: Emerging global opportunistic pathogens

PubMed Central

2018-01-01

ABSTRACT Non-fermenting Gram-negative bacteria are problematic in clinical locations, being one of the most prevalent causes of nosocomial infections. Many of these non-fermenting Gram-negative bacteria are opportunistic pathogens that affect patients that are suffering with underlying medical conditions and diseases. Brevundimonas spp., in particular Brevundimonas diminuta and Brevundimonas vesicularis, are a genus of non-fermenting Gram-negative bacteria considered of minor clinical importance. Forty-nine separate instances of infection relating to Brevundimonas spp were found in the scientific literature along with two pseudo-infections. The majority of these instances were infection with Brevundimonas vesicularis (thirty-five cases – 71%). The major condition associated with Brevundimonas spp infection was bacteraemia with seventeen individual cases/outbreaks (35%). This review identified forty-nine examples of Brevundimonas spp. infections have been discussed in the literature. These findings indicate that infection review programs should consider investigation of possible Brevundimonas spp outbreaks if these bacteria are clinically isolated in more than one patient. PMID:29484917

Caspase-11 cleaves gasdermin D for non-canonical inflammasome signalling.

PubMed

Kayagaki, Nobuhiko; Stowe, Irma B; Lee, Bettina L; O'Rourke, Karen; Anderson, Keith; Warming, Søren; Cuellar, Trinna; Haley, Benjamin; Roose-Girma, Merone; Phung, Qui T; Liu, Peter S; Lill, Jennie R; Li, Hong; Wu, Jiansheng; Kummerfeld, Sarah; Zhang, Juan; Lee, Wyne P; Snipas, Scott J; Salvesen, Guy S; Morris, Lucy X; Fitzgerald, Linda; Zhang, Yafei; Bertram, Edward M; Goodnow, Christopher C; Dixit, Vishva M

2015-10-29

Intracellular lipopolysaccharide from Gram-negative bacteria including Escherichia coli, Salmonella typhimurium, Shigella flexneri, and Burkholderia thailandensis activates mouse caspase-11, causing pyroptotic cell death, interleukin-1β processing, and lethal septic shock. How caspase-11 executes these downstream signalling events is largely unknown. Here we show that gasdermin D is essential for caspase-11-dependent pyroptosis and interleukin-1β maturation. A forward genetic screen with ethyl-N-nitrosourea-mutagenized mice links Gsdmd to the intracellular lipopolysaccharide response. Macrophages from Gsdmd(-/-) mice generated by gene targeting also exhibit defective pyroptosis and interleukin-1β secretion induced by cytoplasmic lipopolysaccharide or Gram-negative bacteria. In addition, Gsdmd(-/-) mice are protected from a lethal dose of lipopolysaccharide. Mechanistically, caspase-11 cleaves gasdermin D, and the resulting amino-terminal fragment promotes both pyroptosis and NLRP3-dependent activation of caspase-1 in a cell-intrinsic manner. Our data identify gasdermin D as a critical target of caspase-11 and a key mediator of the host response against Gram-negative bacteria.

VIP as a potential therapeutic agent in gram negative sepsis.

PubMed

Ibrahim, Hiba; Barrow, Paul; Foster, Neil

2012-12-01

Gram negative sepsis remains a high cause of mortality and places a great burden on public health finance in both the developed and developing world. Treatment of sepsis, using antibiotics, is often ineffective since pathology associated with the disease occurs due to dysregulation of the immune system (failure to return to steady state conditions) which continues after the bacteria, which induced the immune response, have been cleared. Immune modulation is therefore a rational approach to the treatment of sepsis but to date no drug has been developed which is highly effective, cheap and completely safe to use. One potential therapeutic agent is VIP, which is a natural peptide and is highly homologous in all vertebrates. In this review we will discuss the effect of VIP on components of the immune system, relevant to gram negative sepsis, and present data from animal models. Furthermore we will hypothesise on how these studies could be improved in future and speculate on the possible different ways in which VIP could be used in clinical medicine.

Discovery and Optimization of Indolyl-Containing 4-Hydroxy-2-Pyridone Type II DNA Topoisomerase Inhibitors Active against Multidrug Resistant Gram-negative Bacteria.

PubMed

Gerasyuto, Aleksey I; Arnold, Michael A; Wang, Jiashi; Chen, Guangming; Zhang, Xiaoyan; Smith, Sean; Woll, Matthew G; Baird, John; Zhang, Nanjing; Almstead, Neil G; Narasimhan, Jana; Peddi, Srinivasa; Dumble, Melissa; Sheedy, Josephine; Weetall, Marla; Branstrom, Arthur A; Prasad, J V N; Karp, Gary M

2018-05-14

There exists an urgent medical need to identify new chemical entities (NCEs) targeting multidrug resistant (MDR) bacterial infections, particularly those caused by Gram-negative pathogens. 4-Hydroxy-2-pyridones represent a novel class of nonfluoroquinolone inhibitors of bacterial type II topoisomerases active against MDR Gram-negative bacteria. Herein, we report on the discovery and structure-activity relationships of a series of fused indolyl-containing 4-hydroxy-2-pyridones with improved in vitro antibacterial activity against fluoroquinolone resistant strains. Compounds 6o and 6v are representative of this class, targeting both bacterial DNA gyrase and topoisomerase IV (Topo IV). In an abbreviated susceptibility screen, compounds 6o and 6v showed improved MIC 90 values against Escherichia coli (0.5-1 μg/mL) and Acinetobacter baumannii (8-16 μg/mL) compared to the precursor compounds. In a murine septicemia model, both compounds showed complete protection in mice infected with a lethal dose of E. coli.

The Importance of Mean Neutrophil Volume (MNV) Values in Differential Diagnosis of Bacterial Sepsis.

PubMed

Şafak, Birol; Baykan, Özgür; Kılınç, Osman; Özer Yıldırım, Diğdem

2017-01-01

Bloodstream infections are a significant cause of morbidity and mortality in hospitalized patients. Blood cultures and other laboratory tests are used for diagnosis. Among these tests, the mean neutrophil volume (MNV) value is reported as a potential indicator that supports the diagnosis of sepsis. Our study identified the MNV values of patients via microorganisms cultivated from blood cultures and examined the role of these MNV values in the early diagnosis of bloodstream infections. Our study surveyed retrospectively 148 adult patient blood culture samples that had been sent to our laboratory. BACTEC 9050 (Becton Dickinson, USA) and BACTEC FX 40 (Becton Dickinson, USA) devices were used in the blood culture isolation procedures. The average MNV value was found to be 159.0 (+11.3) in patients whose sepsis originated from Gram-negative bacteria, and the average MNV value was measured as 152.4 (+14.5) among patients whose sepsis originated from Gram-positive bacteria. When comparing groups of patients having Gram-negative bacteria and patients having Gram-positive bacteria, a statistically significant difference (p = 0.041) in the MNV values was observed. The MNV value was found to be statistically significant in discrimination of Gram-negative and Gram-positive sepsis. Considering these findings, measuring the MNV values can help initiate proper antibiotic treatment more quickly, and we think that this will help lower the mortality rate. However, these findings should be supported with further studies. Copyright © 2017 National Medical Association. Published by Elsevier Inc. All rights reserved.

Polymicrobial Infective Endocarditis: Clinical Features and Prognosis

PubMed Central

García-Granja, Pablo Elpidio; López, Javier; Vilacosta, Isidre; Ortiz-Bautista, Carlos; Sevilla, Teresa; Olmos, Carmen; Sarriá, Cristina; Ferrera, Carlos; Gómez, Itziar; Román, José Alberto San

2015-01-01

Abstract To describe the profile of left-sided polymicrobial endocarditis (PE) and to compare it with monomicrobial endocarditis (ME). Among 1011 episodes of left-sided endocarditis consecutively diagnosed in 3 tertiary centers, between January 1, 1996 and December 31, 2014, 60 were polymicrobial (5.9%), 821 monomicrobial (81.7%), and in 123 no microorganism was detected (12.2%). Seven patients (0.7%) were excluded from the analysis because contamination of biologic tissue could not be discarded. The authors described the clinical, microbiologic, echocardiographic, and outcome of patients with PE and compared it with ME. Mean age was 64 years SD 16 years, 67% were men and 30% nosocomial. Diabetes mellitus (35%) were the most frequent comorbidities, fever (67%) and heart failure (43%) the most common symptoms at admission. Prosthetic valves (50%) were the most frequent infection location and coagulase-negative Staphylococci (48%) and enterococci (37%) the leading etiologies. The most repeated combination was coagulase-negative Staphylococci with enterococci (n = 9). Polymicrobial endocarditis appeared more frequently in patients with underlying disease (70% versus 56%, P = 0.036), mostly diabetics (35% versus 24%, P = 0.044) with previous cardiac surgery (15% versus 8% P = 0.049) and prosthetic valves (50% versus 37%, P = 0.038). Coagulase-negative Staphylococci, enterococci, Gram-negative bacilli, anaerobes, and fungi were more frequent in PE. No differences on age, sex, symptoms, need of surgery, and in-hospital mortality were detected. Polymicrobial endocarditis represents 5.9% of episodes of left-sided endocarditis in our series. Despite relevant demographic and microbiologic differences between PE and ME, short-term outcome is similar. PMID:26656328

Four-year analysis of microbial aetiology and antimicrobial sensitivity patterns of peritoneal-dialysis related peritonitis in a tertiary care facility.

PubMed

Persy, B; Ieven, M

2013-01-01

Peritonitis related to peritoneal dialysis increases morbidity and mortality and is the main reason for switching to haemodialysis. In this study, we analysed the dialysate of 164 peritoneal dialysis patients that was sent to our laboratory between January 2005 and August 2009. There were 196 peritonitis episodes identified in 78 patients. For all episodes, microbial aetiologies and in-vitro antimicrobial sensitivities were determined in addition to parameters such as the leukocyte count (WBC) and the result of the Gram stain. Results in children were analysed separately because of previously described differences in aetiology. In both groups, Gram positives were most commonly isolated, followed by Gram negatives and fungi or yeasts. In children, the proportion of coagulase-negative staphylococci compared to S. aureus is smaller than in adults. Gram stain showed predominant morphotypes concordant with culture results in 28% of episodes. A significant difference in WBC count was found between culture-positive (mean=3117 x 10(9)/L) and culture-negative (mean=981 x 10(9)/L) episodes in adults (p=0.001). The WBC count in episodes caused exclusively by CNS (mean=1502x10(9)/L) was also on average significantly lower (p=0.001) compared to all culture positive episodes. Resistance to methicillin was registered in 33% of cultures, positive for staphylococci. All Gram-positives were sensitive to vancomycin. Coverage of Gram-negatives by ceftazidim and quinolones was excellent (89%). Based on local sensitivity data and known characteristics of antimicrobials, a first-line empirical combination of intraperitoneal vancomycin with orally administered ciprofloxacin seems indicated in our population. Pathogens of positive aerobic cultures were sensitive in-vitro to their combined antimicrobial effect in 90% of cases.

Potential strategies for the eradication of multidrug-resistant Gram-negative bacterial infections.

PubMed

Huwaitat, Rawan; McCloskey, Alice P; Gilmore, Brendan F; Laverty, Garry

2016-07-01

Antimicrobial resistance is one of the leading threats to society. The increasing burden of multidrug-resistant Gram-negative infection is particularly concerning as such bacteria are demonstrating resistance to nearly all currently licensed therapies. Various strategies have been hypothesized to treat multidrug-resistant Gram-negative infections including: targeting the Gram-negative outer membrane; neutralization of lipopolysaccharide; inhibition of bacterial efflux pumps and prevention of protein folding. Silver and silver nanoparticles, fusogenic liposomes and nanotubes are potential strategies for extending the activity of licensed, Gram-positive selective, antibiotics to Gram-negatives. This may serve as a strategy to fill the current void in pharmaceutical development in the short term. This review outlines the most promising strategies that could be implemented to solve the threat of multidrug-resistant Gram-negative infections.

Transient sensitivity to nisin in cold-shocked Gram negatives.

PubMed

Boziaris, I S; Adams, M R

2000-09-01

Rapid chilling in the presence of nisin caused a dose-dependent reduction in the populations of several Gram-negative bacteria, despite the fact that appreciable structural injury to the outer membrane was not detected. Pseudomonas aeruginosa was most affected, followed by Pseudomonas fragi, Salmonella enteritidis PT4, PT7 and Escherichia coli, respectively. Addition of nisin after the chilling treatment had no effect. The results are ascribed to a transient susceptibility caused by phase changes in the lipids associated with the outer membrane, which are rapidly reversed when the cells return to higher temperatures. Combinations of chilling shock, nisin and EDTA gave much lower reductions of Salmonella and Pseudomonas on chicken skin in comparison with broths. This is attributed to a buffering of the temperature shock experienced by adherent bacteria and binding of the nisin by food particles.

Rapid Discrimination of Gram-Positive and Gram-Negative Bacteria in Liquid Samples by Using NaOH-Sodium Dodecyl Sulfate Solution and Flow Cytometry

PubMed Central

Wada, Atsushi; Kono, Mari; Kawauchi, Sawako; Takagi, Yuri; Morikawa, Takashi; Funakoshi, Kunihiro

2012-01-01

Background For precise diagnosis of urinary tract infections (UTI), and selection of the appropriate prescriptions for their treatment, we explored a simple and rapid method of discriminating gram-positive and gram-negative bacteria in liquid samples. Methodology/Principal Findings We employed the NaOH-sodium dodecyl sulfate (SDS) solution conventionally used for plasmid extraction from Escherichia coli and the automated urine particle analyzer UF-1000i (Sysmex Corporation) for our novel method. The NaOH-SDS solution was used to determine differences in the cell wall structures between gram-positive and gram-negative bacteria, since the tolerance to such chemicals reflects the thickness and structural differences of bacterial cell walls. The UF-1000i instrument was used as a quantitative bacterial counter. We found that gram-negative bacteria, including E. coli, in liquid culture could easily be lysed by direct addition of equal volumes of NaOH-SDS solution. In contrast, Enterococcus faecalis, which is a gram-positive bacterium, could not be completely lysed by the solution. We then optimized the reaction time of the NaOH-SDS treatment at room temperature by using 3 gram-positive and 4 gram-negative bacterial strains and determined that the optimum reaction time was 5 min. Finally, in order to evaluate the generalizability of this method, we treated 8 gram-positive strains and 8 gram-negative strains, or 4 gram-positive and 4 gram-negative strains incubated in voluntary urine from healthy volunteers in the same way and demonstrated that all the gram-positive bacteria were discriminated quantitatively from gram negative bacteria using this method. Conclusions/Significance Using our new method, we could easily discriminate gram-positive and gram-negative bacteria in liquid culture media within 10 min. This simple and rapid method may be useful for determining the treatment course of patients with UTIs, especially for those without a prior history of UTIs. The method may be easily applied in order to obtain additional information for clinical prescriptions from bacteriuria. PMID:23077549

Reactivation of Latent Tuberculosis: Variations on the Cornell Murine Model

PubMed Central

Scanga, Charles A.; Mohan, V. P.; Joseph, Heather; Yu, Keming; Chan, John; Flynn, JoAnne L.

1999-01-01

Mycobacterium tuberculosis causes active tuberculosis in only a small percentage of infected persons. In most cases, the infection is clinically latent, although immunosuppression can cause reactivation of a latent M. tuberculosis infection. Surprisingly little is known about the biology of the bacterium or the host during latency, and experimental studies on latent tuberculosis suffer from a lack of appropriate animal models. The Cornell model is a historical murine model of latent tuberculosis, in which mice infected with M. tuberculosis are treated with antibiotics (isoniazid and pyrazinamide), resulting in no detectable bacilli by organ culture. Reactivation of infection during this culture-negative state occurred spontaneously and following immunosuppression. In the present study, three variants of the Cornell model were evaluated for their utility in studies of latent and reactivated tuberculosis. The antibiotic regimen, inoculating dose, and antibiotic-free rest period prior to immunosuppression were varied. A variety of immunosuppressive agents, based on immunologic factors known to be important to control of acute infection, were used in attempts to reactivate the infection. Although reactivation of latent infection was observed in all three variants, these models were associated with characteristics that limit their experimental utility, including spontaneous reactivation, difficulties in inducing reactivation, and the generation of altered bacilli. The results from these studies demonstrate that the outcome of Cornell model-based studies depends critically upon the parameters used to establish the model. PMID:10456896

Are Fluoroquinolones Appropriate for the Treatment of Extended-Spectrum β-Lactamase-Producing Gram-Negative Bacilli?

PubMed Central

Wiener, Emily S.; Heil, Emily L.; Hynicka, Lauren M.; Johnson, J. Kristie

2015-01-01

Objective: To review the data analyzing the role of fluoroquinolones in the treatment of extended-spectrum β-lactamase (ESBL)-producing infections and rates and methods of co-transmission of resistance. Data Sources: A MEDLINE literature search was performed using the search terms extended-spectrum beta-lactamase, fluoroquinolone, ciprofloxacin, levofloxacin, plasmid transmission, and resistance from 1996 to June 2015. Additional references were identified from a review of literature citations. Study Selection and Data Extraction: All English-language retrospective studies, prospective studies, and meta-analyses assessing efficacy of fluoroquinolone use in ESBL infections, assessing methods of resistance transmission, or analyzing patient risk factors were reviewed. Data Synthesis: A total of 18 studies that analyzed fluoroquinolone resistance and association to ESBL producing bacteria from either molecular or clinical perspectives were idenitifed. Four studies evaluated the genetic association between ESBL transmission and fluoroquinolone resistance. Plasmid mediated quinolone resistance was found in higher rates in ESBL-producing bacteria. Numerous studies analyzed the risk factors of co-occurring resistance identifying nosocomial acquired infections, recent hospitalization, long-term care facility residence, and intensive care unit stay as the most common. Conclusive clinical data are lacking; however, a meta-analysis showed fluoroquinolones had higher odds of all-cause mortality when used empirically to treat ESBL bacteremia compared with carbapenems. Conclusions: Fluoroquinolone resistance may be co-transmitted in ESBL-producing Enterobacteriaceae. There are limited data on the efficacy for fluoroquinolones in the treatment of ESBL-producing infections. Additional prospective trials are needed to definitively determine the role of fluoroquinolones in ESBL infections.

Escherichia coli Isolated from Urinary Tract Infections of Lebanese Patients between 2005 and 2012: Epidemiology and Profiles of Resistance.

PubMed

Daoud, Ziad; Salem Sokhn, Elie; Masri, Khalil; Matar, Ghassan M; Doron, Shira

2015-01-01

The early treatment of urinary tract infections (UTIs) is directly related to decrease in morbidity, which makes the empirical treatment of great importance. Recently, beta lactamases of several types have emerged as significant mechanisms of resistance in Gram-negative bacilli, especially Escherichia coli. Our aim was to study the urinary E. coli isolated from Lebanese patients and to characterize their mechanisms of resistance. The study analyzed data between 2005 and 2012 of UTIs caused by E. coli. The mechanisms of resistance were characterized by phenotypic and genotypic methods and the pulsed-field gel electrophoresis (PFGE) was used to determine the different bacterial clusters. As expected, the highest incidence was observed with E. coli (60.53-73.98%) followed by K. pneumoniae (5.32-8.33%). ICU isolates were constantly associated with the lowest rates of susceptibility to extended-spectrum cephalosporins, ciprofloxacin, as well as most of the tested antibiotics. A 100% occurrence of CTX-M in extended-spectrum β-lactamase (ESBL)-producing isolates was recorded, followed by TEM, SHV, and OXA. In addition, 15.9% harbored 4 different ESBL enzymes and only 13 isolates (14.8%) harbored only one enzyme (CTX-M). Over the years, the simultaneous susceptibility of E. coli to ceftazidime and ciprofloxacin decreased from 62.5% in 2006 to 48.7% in 2012. PFGE results demonstrated that 10 clusters were 32 generated, denoting diversity among detected isolates. Understanding the epidemiology of resistance is 33 instrumental for the implementation of recommendations for the management of antimicrobials, infection 34 control measures, as well as active surveillance and antimicrobial stewardship.

Inhibition of Fosfomycin Resistance Protein FosA by Phosphonoformate (Foscarnet) in Multidrug-Resistant Gram-Negative Pathogens

PubMed Central

Ito, Ryota; Tomich, Adam D.; McElheny, Christi L.; Mettus, Roberta T.; Sluis-Cremer, Nicolas

2017-01-01

ABSTRACT FosA proteins confer fosfomycin resistance to Gram-negative pathogens via glutathione-mediated modification of the antibiotic. In this study, we assessed whether inhibition of FosA by sodium phosphonoformate (PPF) (foscarnet), a clinically approved antiviral agent, would reverse fosfomycin resistance in representative Gram-negative pathogens. The inhibitory activity of PPF against purified recombinant FosA from Escherichia coli (FosA3), Klebsiella pneumoniae (FosAKP), Enterobacter cloacae (FosAEC), and Pseudomonas aeruginosa (FosAPA) was determined by steady-state kinetic measurements. The antibacterial activity of PPF against FosA in clinical strains of these species was evaluated by susceptibility testing and time-kill assays. PPF increased the Michaelis constant (Km) for fosfomycin in a dose-dependent manner, without affecting the maximum rate (Vmax) of the reaction, for all four FosA enzymes tested, indicating a competitive mechanism of inhibition. Inhibitory constant (Ki) values were 22.6, 35.8, 24.4, and 56.3 μM for FosAKP, FosAEC, FosAPA, and FosA3, respectively. Addition of clinically achievable concentrations of PPF (∼667 μM) reduced the fosfomycin MICs by ≥4-fold among 52% of the K. pneumoniae, E. cloacae, and P. aeruginosa clinical strains tested and led to a bacteriostatic or bactericidal effect in time-kill assays among representative strains. PPF inhibits FosA activity across Gram-negative species and can potentiate fosfomycin activity against the majority of strains with chromosomally encoded fosA. These data suggest that PPF may be repurposed as an adjuvant for fosfomycin to treat infections caused by some FosA-producing, multidrug-resistant, Gram-negative pathogens. PMID:28993329

Identification and antimicrobial resistance of pathogens in neonatal septicemia in China-A meta-analysis.

PubMed

Li, Jing-Yang; Chen, Shang-Qin; Yan, Yan-Yan; Hu, Ying-Ying; Wei, Jia; Wu, Qiu-Ping; Lin, Zhen-Lang; Lin, Jing

2018-06-01

The purpose of this study was to analyze the distribution and antimicrobial resistance of common bacterial pathogens causing neonatal septicemia based on a systematic review of published studies in China. Articles on neonatal sepsis published in the Chinese literature from 2009 to 2014 were identified according to the inclusion and exclusion criteria. Data were extracted and analyzed using Comprehensive Meta-Analysis software. A total of 71 studies were included, in which a total of 8080 bacterial species were isolated from culture-positive blood samples. The pooled distribution rates of common bacterial pathogens were as follows: Staphylococcus 67.1% (95% confidence interval (CI) 63.3-70.6%), Enterococcus 4.1% (95% CI 3.5-4.8%), Streptococcus 2.3% (95% CI 1.6-3.2%), Escherichia coli 7.4% (95% CI 6.4-8.7%), Klebsiella 6.5% (95% CI 5.2-8.2%), Enterobacterium 2.3% (95% CI 1.9-2.8%), Acinetobacter 1.6% (95% CI 1.3-2.0%), Pseudomonas 1.7% (95% CI 1.3-2.2%). Among the Staphylococcus aureus strains isolated, more than 60% were methicillin-resistant (MRSA). In addition, over 50% of the Gram-negative isolates, including Escherichia and Klebsiella, were resistant to the commonly used third-generation cephalosporins. Most of the Gram-positive and Gram-negative bacteria isolated were sensitive to aminoglycosides, especially amikacin. It is concluded that Staphylococcus, especially coagulase-negative Staphylococcus, continues to be the principal organism responsible for neonatal septicemia in China; Enterobacteriaceae are common among the Gram-negative isolates. Significant numbers of MRSA and multidrug-resistant Gram-negative bacteria are being isolated as pathogens responsible for neonatal septicemia in China. Copyright © 2018 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Dietary Heme Alters Microbiota and Mucosa of Mouse Colon without Functional Changes in Host-Microbe Cross-Talk

PubMed Central

van Doorn, Gerdien M.; Rijnierse, Anneke; van den Bogert, Bartholomeus; Müller, Michael; Dekker, Jan; Kleerebezem, Michiel; van der Meer, Roelof

2012-01-01

Colon cancer is a major cause of cancer deaths in Western countries and is associated with diets high in red meat. Heme, the iron-porphyrin pigment of red meat, induces cytotoxicity of gut contents which injures surface cells leading to compensatory hyperproliferation of crypt cells. This hyperproliferation results in epithelial hyperplasia which increases the risk of colon cancer. In humans, a high red-meat diet increases Bacteroides spp in feces. Therefore, we simultaneously investigated the effects of dietary heme on colonic microbiota and on the host mucosa of mice. Whole genome microarrays showed that heme injured the colonic surface epithelium and induced hyperproliferation by changing the surface to crypt signaling. Using 16S rRNA phylogenetic microarrays, we investigated whether bacteria play a role in this changed signaling. Heme increased Bacteroidetes and decreased Firmicutes in colonic contents. This shift was most likely caused by a selective susceptibility of Gram-positive bacteria to heme cytotoxic fecal water, which is not observed for Gram-negative bacteria, allowing expansion of the Gram-negative community. The increased amount of Gram-negative bacteria most probably increased LPS exposure to colonocytes, however, there is no appreciable immune response detected in the heme-fed mice. There was no functional change in the sensing of the bacteria by the mucosa, as changes in inflammation pathways and Toll- like receptor signaling were not detected. This unaltered host-microbe cross-talk indicates that the changes in microbiota did not play a causal role in the observed hyperproliferation and hyperplasia. PMID:23239972

Corky root of lettuce caused by strains of a gram-negative bacterium from muck soils of Florida, new york, and wisconsin.

PubMed

van Bruggen, A H; Brown, P R; Jochimsen, K N

1989-10-01

Slow-growing bacteria similar to the bacterium causing lettuce corky root (CR) in California (strain CA1) were isolated from muck soils of Florida, New York, and Wisconsin, using lettuce seedlings as bait. All strains were tested for reaction with polyclonal antibodies produced against strain CA1 and for pathogenicity on CR-susceptible (Salinas) and CR-resistant (Green Lake) lettuce cultivars in a greenhouse. Five strains from Florida, three from New York, and three from Wisconsin induced severe CR symptoms on Salinas and mild symptoms on Green Lake. All strains were gram-negative, aerobic, oxidase positive, and catalase positive and reduced nitrate to ammonia. Whole-cell fatty acid compositions were similar for all strains and resembled that of Pseudomonas paucimobilis. Since this fatty acid pattern is unique, it is suggested that CR of lettuce is caused by strains of the same bacterium in Florida, New York, Wisconsin, and California.