scholarly journals Bordetella holmesii Contamination of Platelet Concentrates: Revisiting the Definition of a Positive Culture

2018 ◽  
Vol 56 (12) ◽  
Author(s):  
Marc Cloutier ◽  
Marie-Ève Nolin ◽  
Hana Daoud ◽  
Annie Jacques ◽  
Marie Joëlle de Grandmont ◽  
...  
Keyword(s):  
Positive Culture ◽  
Blood Products ◽  
Platelet Concentrates ◽  
Bacterial Strains ◽  
Culture Bottle ◽  
Decision Algorithm ◽  
Content Type ◽  
Infectious Risk ◽  
Definition Of

ABSTRACT Bacterial contamination remains the most important infectious risk of platelet transfusion. After an initially positive result, a second test is performed on the blood products and the initial culture bottle to confirm the contamination. Based on the blood center's decision algorithm used, results can be either confirmed negative, positive, or indeterminate, or be unconfirmed or discordant. Here, we report the first cases of platelet concentrates contaminated with Bordetella holmesii. The in vitro growth characteristics of this unusual contaminant in platelet concentrate were investigated. Two B. holmesii strains isolated from platelet concentrates, as well as a control strain (Serratia marcescens), were spiked into platelet concentrates (PCs) at 1 and 10 CFU/ml. PCs were stored at 20 to 24°C under agitation. Samples were collected on days 2, 3, 4, and 7 for colony count and for bacterial screening using the BacT/Alert 3D system. Two PCs were detected as being positive for B. holmesii. However, recultures were negative. In vitro, B. holmesii did not grow but remained detectable in PCs. Its viability diminished rapidly in contact with human plasma. Upon screening using the BacT/Alert 3D system, the majority of products spiked with B. holmesii were negative. This is the first description of PCs contaminated with B. holmesii. This bacterium survives in blood products and remains dormant at low concentrations in blood products stored at room temperature, thus making difficult its detection with the BacT/Alert 3D system. The present definition of a true-positive culture of PCs may be overly restrictive for certain bacterial strains.

scholarly journals Drug Susceptibility Distributions of Mycobacterium chimaera and Other Nontuberculous Mycobacteria

2021 ◽  
Vol 65 (5) ◽  
Author(s):  
Bettina Schulthess ◽  
Daniel Schäfle ◽  
Nicole Kälin ◽  
Tamara Widmer ◽  
Peter Sander
Keyword(s):  
Antibiotic Susceptibility ◽  
Nontuberculous Mycobacteria ◽  
Drug Susceptibility ◽  
Species Differentiation ◽  
Content Type ◽  
Susceptibility Data ◽  
Clinical Breakpoints ◽  
Antibiotic Susceptibility Patterns ◽  
Definition Of

ABSTRACT Recent outbreaks of cardiac surgery-associated Mycobacterium chimaera infections have highlighted the importance of species differentiation within the Mycobacterium avium complex and pointed to a lack of antibiotic susceptibility data for M. chimaera. Using the MGIT 960/EpiCenter TB eXiST platform, we have determined antibiotic susceptibility patterns of 48 clinical M. chimaera isolates and 139 other nontuberculous mycobacteria, including 119 members of the M. avium complex and 20 Mycobacterium kansasii isolates toward clofazimine and other drugs used to treat infections with slow-growing nontuberculous mycobacteria (NTM). MIC50, MIC90, and tentative epidemiological cutoff (ECOFF) values for clofazimine were 0.5 mg/liter, 1 mg/liter, and 2 mg/liter, respectively, for M. chimaera. Comparable values were observed for other M. avium complex members, whereas lower MIC50 (≤0.25 mg/liter), MIC90 (0.5 mg/liter), and ECOFF (1 mg/liter) values were found for M. kansasii. Susceptibility to clarithromycin, ethambutol, rifampin, rifabutin, amikacin, moxifloxacin, and linezolid was in general similar for M. chimaera and other members of the M. avium complex, but increased for M. kansasii. The herein determined MIC distributions, MIC90, and ECOFF values of clofazimine for M. chimaera and other NTM provide the basis for the definition of clinical breakpoints. Further studies are needed to establish correlation of in vitro susceptibility and clinical outcome.

scholarly journals Tagging Frogs with Passive Integrated Transponders Causes Disruption of the Cutaneous Bacterial Community and Proliferation of Opportunistic Fungi

2014 ◽  
Vol 80 (15) ◽  
pp. 4779-4784 ◽  
Author(s):  
Rachael E. Antwis ◽  
Gerardo Garcia ◽  
Andrea L. Fidgett ◽  
Richard F. Preziosi
Keyword(s):  
Bacterial Community ◽  
Microbial Communities ◽  
Bacterial Communities ◽  
Pathogenic Fungus ◽  
Fold Increase ◽  
Bacterial Strains ◽  
Opportunistic Fungi ◽  
Content Type ◽  
Pit Tagging

ABSTRACTSymbiotic bacterial communities play a key role in protecting amphibians from infectious diseases including chytridiomycosis, caused by the pathogenic fungusBatrachochytrium dendrobatidis. Events that lead to the disruption of the bacterial community may have implications for the susceptibility of amphibians to such diseases. Amphibians are often marked both in the wild and in captivity for a variety of reasons, and although existing literature indicates that marking techniques have few negative effects, the response of cutaneous microbial communities has not yet been investigated. Here we determine the effects of passive integrated transponder (PIT) tagging on culturable cutaneous microbial communities of captive Morelet's tree frogs (Agalychnis moreletii) and assess the isolated bacterial strains for anti-B. dendrobatidisactivityin vitro. We find that PIT tagging causes a major disruption to the bacterial community associated with the skin of frogs (∼12-fold increase in abundance), as well as a concurrent proliferation in resident fungi (up to ∼200-fold increase). Handling also caused a disruption the bacterial community, although to a lesser extent than PIT tagging. However, the effects of both tagging and handling were temporary, and after 2 weeks, the bacterial communities were similar to their original compositions. We also identify two bacterial strains that inhibitB. dendrobatidis, one of which increased in abundance on PIT-tagged frogs at 1 day postmarking, while the other was unaffected. These results show that PIT tagging has previously unobserved consequences for cutaneous microbial communities of frogs and may be particularly relevant for studies that intend to use PIT tagging to identify individuals involved in trials to develop probiotic treatments.

scholarly journals Legionella steelei sp. nov., isolated from human respiratory specimens in California, USA, and South Australia

2012 ◽  
Vol 62 (Pt_8) ◽  
pp. 1766-1771 ◽  
Author(s):  
Paul H. Edelstein ◽  
Martha A. Edelstein ◽  
Lisa J. Shephard ◽  
Kevin W. Ward ◽  
Rodney M. Ratcliff
Keyword(s):  
A549 Cells ◽  
South Australia ◽  
Bacterial Strains ◽  
Poor Growth ◽  
Content Type ◽  
Link Type ◽  
Fluorescent Pigment ◽  
Solid Media ◽  
Yeast Extract Medium

Legionella -like bacteria were isolated from the respiratory tract of two patients in California, USA, and South Australia, but were not thought to cause disease. These bacteria, strains F2632 and IMVS-3376T, were found to have identical Legionella macrophage infectivity potentiator (mip) gene sequences and were therefore further characterized to determine their genetic and phenotypic relatedness and properties. Both of these Gram-negative-staining bacterial strains grew on buffered charcoal yeast extract medium, were cysteine auxotrophs and made a characteristic diffusible bright yellow fluorescent pigment, with one strain making a late appearing colony-bound blue–white fluorescent pigment. The optimal in vitro growth temperature was 35 °C, with very poor growth at 37 °C in broth or on solid media. There was no growth in human A549 cells at either 35 or 37 °C, but excellent growth in Acanthamoeba castellani at 30 °C and poorer growth at 35 °C. Phylogenetic analysis of these bacteria was performed by sequence analysis of 16S rRNA, mip, ribonuclease P, ribosomal polymerase B and zinc metalloprotease genes. These studies confirmed that the new strains represented a single novel species of the genus Legionella for which the name Legionella steelei sp. nov. is proposed. The type strain is IMVS-3376T ( = IMVS 3113T = ATCC BAA-2169T).

scholarly journals Atypical Bacterial Growth within Units of Platelets Challenges Transfusion Medicine Dogma

2018 ◽  
Vol 56 (12) ◽  
Author(s):  
Eric A. Gehrie
Keyword(s):  
Bacterial Contamination ◽  
Current Issue ◽  
Safety Concern ◽  
Positive Culture ◽  
Storage Period ◽  
Transfusion Medicine ◽  
Platelet Concentrates ◽  
Content Type ◽  
Platelet Storage ◽  
Positive Culture Result

ABSTRACT Although transfusion-transmitted bacterial infection is relatively rare, mitigation of bacterial contamination of platelet units is arguably the top current transfusion-related safety concern. Several different technologies have been employed to detect or neutralize bacteria in platelet concentrates. However, studies of the efficacy of these systems have been hampered by problematic definitions of what represents a “true-positive” versus a “false-positive” culture result. In the current issue of the Journal of Clinical Microbiology (M. Cloutier, M.-È. Nolin, H. Daoud, A. Jacques, M. J. de Grandmont, É Ducas, G. Delage, and L. Thibault, J Clin Microbiol 56:e01105-18, 2018, https://doi.org/10.1128/JCM.01105-18), it was demonstrated that the growth of Bordetella holmesii is inhibited by the platelet storage environment, which may explain why the results of initial positive platelet cultures are not always confirmed by subsequent cultures later during the storage period. This important finding is at odds with the generally held belief within the field of transfusion medicine that initially positive platelet cultures that are not confirmed on repeat testing are instrumentation-based false positives. The clinical risk profile of organisms demonstrating storage-related low viability is worthy of further study.

scholarly journals Amikacin-Fosfomycin at a Five-to-Two Ratio: Characterization of Mutation Rates in Microbial Strains Causing Ventilator-Associated Pneumonia and Interactions with Commonly Used Antibiotics

2014 ◽  
Vol 58 (7) ◽  
pp. 3708-3713 ◽  
Author(s):  
A. Bruce Montgomery ◽  
Paul R. Rhomberg ◽  
Tammy Abuan ◽  
Kathie-Anne Walters ◽  
Robert K. Flamm
Keyword(s):  
Fold Increase ◽  
Serial Passage ◽  
Multidrug Resistant ◽  
Adjunctive Treatment ◽  
Ventilator Associated Pneumonia ◽  
Bacterial Strains ◽  
Content Type ◽  
Development Of Resistance

ABSTRACTThe amikacin-fosfomycin inhalation system (AFIS), a combination of antibiotics administered with an in-line nebulizer delivery system, is being developed for adjunctive treatment of ventilator-associated pneumonia (VAP). Thein vitrocharacterization of amikacin-fosfomycin (at a 5:2 ratio) described here included determining resistance selection rates for pathogens that are representative of those commonly associated with VAP (including multidrug-resistant strains) and evaluating interactions with antibiotics commonly used intravenously to treat VAP. Spontaneous resistance to amikacin-fosfomycin (5:2) was not observed for most strains tested (n, 10/14). Four strains had spontaneously resistant colonies (frequencies, 4.25 × 10−8to 3.47 × 10−10), for which amikacin-fosfomycin (5:2) MICs were 2- to 8-fold higher than those for the original strains. After 7 days of serial passage, resistance (>4-fold increase over the baseline MIC) occurred in fewer strains (n, 4/14) passaged in the presence of amikacin-fosfomycin (5:2) than with either amikacin (n, 7/14) or fosfomycin (n, 12/14) alone. Interactions between amikacin-fosfomycin (5:2) and 10 comparator antibiotics in checkerboard testing against 30 different Gram-positive or Gram-negative bacterial strains were synergistic (fractional inhibitory concentration [FIC] index, ≤0.5) for 6.7% (n, 10/150) of combinations tested. No antagonism was observed. Synergy was confirmed by time-kill methodology for amikacin-fosfomycin (5:2) plus cefepime (againstEscherichia coli), aztreonam (againstPseudomonas aeruginosa), daptomycin (againstEnterococcus faecalis), and azithromycin (againstStaphylococcus aureus). Amikacin-fosfomycin (5:2) was bactericidal at 4-fold the MIC for 7 strains tested. The reduced incidence of development of resistance to amikacin-fosfomycin (5:2) compared with that for amikacin or fosfomycin alone, and the lack of negative interactions with commonly used intravenous antibiotics, further supports the development of AFIS for the treatment of VAP.

Supernatant of stored platelets causes lung inflammation and coagulopathy in a novel in vivo transfusion model

Blood ◽  
2010 ◽  
Vol 116 (8) ◽  
pp. 1360-1368 ◽  
Author(s):  
Alexander P. J. Vlaar ◽  
Jorrit J. Hofstra ◽  
Wim Kulik ◽  
Henk van Lenthe ◽  
Rienk Nieuwland ◽  
...  
Keyword(s):  
Acute Lung Injury ◽  
Lung Injury ◽  
Lung Inflammation ◽  
Pulmonary Injury ◽  
Blood Products ◽  
Platelet Concentrates ◽  
Priming Activity ◽  
Neutrophil Priming

Abstract Transfusion-related acute lung injury is suggested to be a “2-hit” event resulting from priming and activation of pulmonary neutrophils. Activation may result from infusion of lysophosphatidylcholines (LysoPCs), which accumulate during storage of blood products. In the present study, we developed a syngeneic in vivo transfusion model to test whether storage of platelet concentrates (PLTs) results in lung injury in healthy rats as well as in a “2-hit” model using lipopolysaccharide-pretreated rats. In addition, the effect of washing of platelets was studied. In healthy rats, transfusion of aged PLTs caused mild lung inflammation. In LPS-pretreated rats, transfusion of aged PLTs, but not fresh PLTs, augmented pulmonary systemic coagulopathy. When PLTs components were transfused separately, supernatant of aged PLTs, but not washed aged platelets, induced pulmonary injury in the “2-hit” model. Supernatants of aged PLTs contained increased concentrations of LysoPCs compared with fresh PLTs, which enhanced neutrophil priming activity in vitro. We conclude that transfusion of aged PLTs induces lung inflammation in healthy rats. In a “2-hit” model, aged PLTs contribute to pulmonary and systemic coagulopathy, which may be mediated by LysoPCs, which accumulate in the supernatant of PLTs during storage.

scholarly journals Ecosystem Screening Approach for Pathogen-Associated Microorganisms Affecting Host Disease

2011 ◽  
Vol 77 (17) ◽  
pp. 6069-6075 ◽  
Author(s):  
Eric Galiana ◽  
Antoine Marais ◽  
Catherine Mura ◽  
Benoît Industri ◽  
Gilles Arbiol ◽  
...  
Keyword(s):  
Microbial Community ◽  
Screening Method ◽  
Surface Shape ◽  
Host Disease ◽  
Content Type ◽  
Mutualistic Interaction ◽  
Definition Of ◽  
Associated Species

ABSTRACTThe microbial community in which a pathogen evolves is fundamental to disease outcome. Species interacting with a pathogen on the host surface shape the distribution, density, and genetic diversity of the inoculum, but the role of these species is rarely determined. The screening method developed here can be used to characterize pathogen-associated species affecting disease. This strategy involves three steps: (i) constitution of the microbial community, using the pathogen as a trap; (ii) community selection, using extracts from the pathogen as the sole nutrient source; and (iii) molecular identification and the screening of isolates focusing on their effects on the growth of the pathogenin vitroand host disease. This approach was applied to a soilborne plant pathogen,Phytophthora parasitica, structured in a biofilm, for screening the microbial community from the rhizosphere ofNicotiana tabacum(the host). Two of the characterized eukaryotes interfered with the oomycete cycle and may affect the host disease. AVorticellaspecies acted through a mutualistic interaction withP. parasitica, disseminating pathogenic material by leaving the biofilm. APhomaspecies established an amensal interaction withP. parasitica, strongly suppressing disease by inhibitingP. parasiticagermination. This screening method is appropriate for all nonobligate pathogens. It allows the definition of microbial species as promoters or suppressors of a disease for a given biotope. It should also help to identify important microbial relationships for ecology and evolution of pathogens.

scholarly journals Oxygen Affects Gut Bacterial Colonization and Metabolic Activities in a Gnotobiotic Cockroach Model

2015 ◽  
Vol 82 (4) ◽  
pp. 1080-1089 ◽  
Author(s):  
Dorothee Tegtmeier ◽  
Claire L. Thompson ◽  
Christine Schauer ◽  
Andreas Brune
Keyword(s):  
Bacterial Colonization ◽  
Anaerobic Bacteria ◽  
Close Relative ◽  
Bacterial Strains ◽  
Fermentation Products ◽  
Content Type ◽  
Facultatively Anaerobic ◽  
Pure Cultures ◽  
Metabolic Activities

ABSTRACTThe gut microbiota of termites and cockroaches represents complex metabolic networks of many diverse microbial populations. The distinct microenvironmental conditions within the gut and possible interactions among the microorganisms make it essential to investigate how far the metabolic properties of pure cultures reflect their activities in their natural environment. We established the cockroachShelfordella lateralisas a gnotobiotic model and inoculated germfree nymphs with two bacterial strains isolated from the guts of conventional cockroaches. Fluorescence microscopy revealed that both strains specifically colonized the germfree hindgut. In diassociated cockroaches, the facultatively anaerobic strain EbSL (a new species ofEnterobacteriaceae) always outnumbered the obligately anaerobic strain FuSL (a close relative ofFusobacterium varium), irrespective of the sequence of inoculation, which showed that precolonization by facultatively anaerobic bacteria does not necessarily favor colonization by obligate anaerobes. Comparison of the fermentation products of the cultures formedin vitrowith those accumulatedin situindicated that the gut environment strongly affected the metabolic activities of both strains. The pure cultures formed the typical products of mixed-acid or butyrate fermentation, whereas the guts of gnotobiotic cockroaches accumulated mostly lactate and acetate. Similar shifts toward more-oxidized products were observed when the pure cultures were exposed to oxygen, which corroborated the strong effects of oxygen on the metabolic fluxes previously observed in termite guts. Oxygen microsensor profiles of the guts of germfree, gnotobiotic, and conventional cockroaches indicated that both gut tissue and microbiota contribute to oxygen consumption and suggest that the oxygen status influences the colonization success.

scholarly journals In Vitro Gut Modeling as a Tool for Adaptive Evolutionary Engineering of Lactiplantibacillus plantarum

mSystems ◽  
2021 ◽  
Vol 6 (2) ◽  
Author(s):  
Julia Isenring ◽  
Annelies Geirnaert ◽  
Alex R. Hall ◽  
Christoph Jans ◽  
Christophe Lacroix ◽  
...  
Keyword(s):  
Gut Microbiota ◽  
Model Organism ◽  
Strain Improvement ◽  
Evolutionary Engineering ◽  
Bacterial Strains ◽  
Content Type ◽  
Colonic Microbiota ◽  
Gut Colonization ◽  
Fermentation Model

ABSTRACT Research and marketing of probiotics demand holistic strain improvement considering both the biotic and abiotic gut environment. Here, we aim to establish the continuous in vitro colonic fermentation model PolyFermS as a tool for adaptive evolutionary engineering. Immobilized fecal microbiota from adult donors were steadily cultivated up to 72 days in PolyFermS reactors, providing a long-term compositional and functional stable ecosystem akin to the donor’s gut. Inoculation of the gut microbiota with immobilized or planktonic Lactiplantibacillus plantarum NZ3400, a derivative of the probiotic model strain WCFS1, led to successful colonization. Whole-genome sequencing of 45 recovered strains revealed mutations in 16 genes involved in signaling, metabolism, transport, and cell surface. Remarkably, mutations in LP_RS14990, LP_RS15205, and intergenic region LP_RS05100<LP_RS05095 were found in recovered strains from different adaptation experiments. Combined addition of the reference strain NZ3400 and each of those mutants to the gut microbiota resulted in increased abundance of the corresponding mutant in PolyFermS microbiota after 10 days, showing the beneficial nature of these mutations. Our data show that the PolyFermS system is a suitable technology to generate adapted mutants for colonization under colonic conditions. Analysis thereof will provide knowledge about factors involved in gut microbiota colonization and persistence. IMPORTANCE Improvement of bacterial strains in regard to specific abiotic environmental factors is broadly used to enhance strain characteristics for processing and product quality. However, there is currently no multidimensional probiotic strain improvement approach for both abiotic and biotic factors of a colon microbiota. The continuous PolyFermS fermentation model allows stable and reproducible continuous cultivation of colonic microbiota and provides conditions akin to the host gut with high control and easy sampling. This study investigated the suitability of PolyFermS for adaptive evolutionary engineering of a probiotic model organism for lactobacilli, Lactiplantibacillus plantarum, to an adult human colonic microbiota. The application of PolyFermS controlled gut microbiota environment led to adaptive evolution of L. plantarum strains for enhanced gut colonization characteristics. This novel tool for strain improvement can be used to reveal relevant factors involved in gut microbiota colonization and develop adapted probiotic strains with improved functionality in the gut.

scholarly journals The Role of Antibiotics in Modulating Virulence in Staphylococcus aureus

2017 ◽  
Vol 30 (4) ◽  
pp. 887-917 ◽  
Author(s):  
Elisabeth Hodille ◽  
Warren Rose ◽  
Binh An Diep ◽  
Sylvain Goutelle ◽  
Gerard Lina ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Immune Response ◽  
Virulence Factors ◽  
Growth Models ◽  
Case Reports ◽  
Suppressive Effect ◽  
Bacterial Strains ◽  
Content Type

SUMMARY Staphylococcus aureus is often involved in severe infections, in which the effects of bacterial virulence factors have great importance. Antistaphylococcal regimens should take into account the different effects of antibacterial agents on the expression of virulence factors and on the host's immune response. A PubMed literature search was performed to select relevant articles on the effects of antibiotics on staphylococcal toxin production and on the host immune response. Information was sorted according to the methods used for data acquisition (bacterial strains, growth models, and antibiotic concentrations) and the assays used for readout generation. The reported mechanisms underlying S. aureus virulence modulation by antibiotics were reviewed. The relevance of in vitro observations is discussed in relation to animal model data and to clinical evidence extracted from case reports and recommendations on the management of toxin-related staphylococcal diseases. Most in vitro data point to a decreased level of virulence expression upon treatment with ribosomally active antibiotics (linezolid and clindamycin), while cell wall-active antibiotics (beta-lactams) mainly increase exotoxin production. In vivo studies confirmed the suppressive effect of clindamycin and linezolid on virulence expression, supporting their utilization as a valuable management strategy to improve patient outcomes in cases of toxin-associated staphylococcal disease.

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