scholarly journals A multi-country study using MALDI-TOF mass spectrometry for rapid identification of Burkholderia pseudomallei

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Wanitda Watthanaworawit ◽  
Tamalee Roberts ◽  
Jill Hopkins ◽  
Ian Gassiep ◽  
Robert Norton ◽  
...  
Keyword(s):  
Burkholderia Cepacia ◽  
Burkholderia Pseudomallei ◽  
Research Unit ◽  
Rapid Identification ◽  
Geographical Differences ◽  
Isolation And Identification ◽  
Maldi Tof ◽  
The Impact ◽  
Vitek Ms

Abstract Background Burkholderia pseudomallei is the bacterial causative agent of melioidosis, a difficult disease to diagnose clinically with high mortality if not appropriately treated. Definitive diagnosis requires isolation and identification of the organism. With the increased adoption of MALDI-TOF MS for the identification of bacteria, we established a method for rapid identification of B. pseudomallei using the Vitek MS, a system that does not currently have B. pseudomallei in its in-vitro diagnostic database. Results A routine direct spotting method was employed to create spectra and SuperSpectra. An initial B. pseudomallei SuperSpectrum was created at Shoklo Malaria Research Unit (SMRU) from 17 reference isolates (46 spectra). When tested, this initial SMRU SuperSpectrum was able to identify 98.2 % (54/55) of Asian isolates, but just 46.7 % (35/75) of Australian isolates. Using spectra (430) from different reference and clinical isolates, two additional SMRU SuperSpectra were created. Using the combination of all SMRU SuperSpectra with seven existing SuperSpectra from Townsville, Australia 119 (100 %) Asian isolates and 31 (100 %) Australian isolates were correctly identified. In addition, no misidentifications were obtained when using these 11 SuperSpectra when tested with 34 isolates of other bacteria including the closely related species Burkholderia thailandensis and Burkholderia cepacia. Conclusions This study has established a method for identification of B. pseudomallei using Vitek MS, and highlights the impact of geographical differences between strains for identification using this technique.

scholarly journals Biological, botanical and chemical alternatives for the control of blackberry (Rubus glaucus Benth.) diseases

2017 ◽  
Vol 70 (2) ◽  
pp. 8169-8176 ◽  
Author(s):  
Oscar Darío Hincapié Echeverri ◽  
Alegría Saldarriaga Cardona ◽  
Cipriano Díaz Diez
Keyword(s):  
Burkholderia Cepacia ◽  
San Antonio ◽  
Block Design ◽  
Experimental Unit ◽  
Trichoderma Asperellum ◽  
Trichoderma Sp ◽  
Chemical Products ◽  
The Impact ◽  
Absolute Control

In order to control the main diseases that affect blackberries (Rubus glaucus Benth.), a research in which 12 treatments to San Antonio ecotype plants originated in vitro was conducted. These treatments were: 1: Trichoderma harzianum+richoderma koningii (Tropical Fungus), 2: Trichoderma sp. (Bioprotection), 3: Trichoderma koningiopsis (Th003 Trichoderma), 4: Trichoderma asperellum (Th034 Trichoderma), 5: Trichoderma asperellum (T-30 Trichoderma), 6: Trichoderma asperellum (T-98 Trichoderma), 7:Burkholderia cepacia (Botrycid), 8: extract of Swinglea glutinosa (Ecoswin), 9: traditional farming treatments (Mancozeb, Propamocarb), 10: chemical products applications (Mancozeb, Mandipropamida, Carbendazim, Propamocar and Metalaxil+Mancozeb) alternated according to the impact of the disease, 11: chemical products applications alternated with organic products according to the suppliers recommendations and presence of the diseases, 12: absolute control, no treatment was applied to the plants. The applications were carried out every 15 days, each plant was an experimental unit and each treatment was made of five experimental units. 12 treatments were made through a RCBD (randomized complete block design) with three repetitions for a total of 15 experimental units per treatment. The assessments were performed every 8 days, and the variables were: number of healthy and sick fruits/treatment, costs/treatment and gross profit. A variance analysis and a Tukey test 5% were made. The best treatments were T11 (rotation of chemicals with biological products), T10 (rotation of chemical products according to the impact of the disease) and T3 (T. koningiopsis); considering the obtained performance/treatment, treatment cost and profit.

scholarly journals A Mouse Model Of Binge Alcohol Consumption andBurkholderiaInfection

2018 ◽  
Author(s):  
Victor Jimenez ◽  
Ryan Moreno ◽  
Erik Settles ◽  
Bart J Currie ◽  
Paul Keim ◽  
...  
Keyword(s):  
Alcohol Consumption ◽  
Binge Drinking ◽  
Burkholderia Pseudomallei ◽  
Alcohol Exposure ◽  
Model Systems ◽  
Phagocytic Cells ◽  
Intracellular Invasion ◽  
The Impact

AbstractBackgroundBinge drinking, a common form of alcohol consumption, is associated with increased mortality and morbidity; yet, its effects on the immune system’s ability to defend against infectious agents are poorly understood.Burkholderia pseudomallei, the causative agent of melioidosis can occur in healthy humans, yet binge alcohol use is progressively being recognized as a major risk factor. Although our previous studies demonstrated that binge alcohol exposure results in reduced alveolar macrophage function and increasedBurkholderiavirulencein vitro, no experimental studies have investigated the outcomes of binge alcohol onBurkholderiaspp. infectionin vivo.Principal FindingsWe used the close genetic relatives ofB. pseudomallei, B. thailandensisE264 andB. vietnamiensis, as useful BSL-2 model systems. Eight-week-old female C57BL/6 mice were administered alcohol comparable to human binge drinking episodes (4.4 g/kg) or PBS intraperitoneally 30 min before a non-lethal intranasal infection. In an initialB. thailandensisinfection (3 x 105), bacteria accumulated in the lungs and disseminated to the spleen in alcohol administered mice only, compared with PBS treated mice at 24 h post-infection (PI). The greatest bacterial load occurred withB. vietnamiensis(1 x 106) in lungs, spleen, and brain tissue by 72 h PI. Pulmonary cytokine expression (TNF-α, GM-CSF) decreased, while splenic cytokine (IL-10) increased in binge drunk mice. Increased lung and brain permeability was observed as early as 2 h post alcohol administrationin vivo.Trans-epithelial electrical resistance (TEER) was significantly decreased, while intracellular invasion of non-phagocytic cells increased with 0.2% v/v alcohol exposurein vitro.ConclusionsOur results indicate that a single binge alcohol dose suppressed innate immune functions and increased the ability of less virulentBurkholderiastrains to disseminate through increased barrier permeability and intracellular invasion of non-phagocytic cells.Author SummaryBurkholderia pseudomalleicauses the disease melioidosis, which occurs in most tropical regions across the globe. Exposure rarely evolves to significant disease in the absence of specific comorbidities, such as binge alcohol intoxication. In susceptible hosts, the disease is primarily manifested as pneumonic melioidosis and can be rapidly fatal if untreated. In this study, we utilizedB. thailandensis, a genetically similar strain toB. pseudomallei, and opportunisticB. vietnamiensis, a known human pathogen that utilizes similar virulence strategies asB. pseudomalleiin immunocompromised and cystic fibrosis patients. The study investigates the impact of a single binge alcohol episode on infectivity and immune responsein vivo. We show that a single binge alcohol episode prior to inhalingBurkholderiaspecies increases bacterial spread to the lungs and brain. We also identify alcohol-induced tissue permeability and epithelial cell invasion as modes of action for greater bacterial spread and survival inside the host. Our results support the public health responses being developed in melioidosis-endemic regions that emphasize the nature of binge drinking as a prime concern, especially around potential times of exposure to environmentalB. pseudomallei.

scholarly journals In Vitro Susceptibilities of Burkholderia mallei in Comparison to Those of Other PathogenicBurkholderia spp

1999 ◽  
Vol 43 (11) ◽  
pp. 2773-2775 ◽  
Author(s):  
D. J. Kenny ◽  
P. Russell ◽  
D. Rogers ◽  
S. M. Eley ◽  
R. W. Titball
Keyword(s):  
Antibiotic Susceptibility ◽  
Burkholderia Cepacia ◽  
Burkholderia Pseudomallei ◽  
Burkholderia Mallei ◽  
Antimicrobial Susceptibilities

ABSTRACT The in vitro antimicrobial susceptibilities of isolates ofBurkholderia mallei to 16 antibiotics were assessed and compared with the susceptibilities of Burkholderia pseudomallei and Burkholderia cepacia. The antibiotic susceptibility profile of B. mallei resembled that ofB. pseudomallei more closely than that of B. cepacia, which corresponds to their similarities in terms of biochemistry, antigenicity, and pathogenicity. Ceftazidime, imipenem, doxycycline, and ciprofloxacin were active against both B. mallei and B. pseudomallei. Gentamicin was active against B. mallei but not against B. pseudomallei. Antibiotics clinically proven to be effective in the treatment of melioidosis may therefore be effective for treating glanders.

scholarly journals Activity of Cysteamine against the Cystic Fibrosis Pathogen Burkholderia cepacia Complex

2016 ◽  
Vol 60 (10) ◽  
pp. 6200-6206 ◽  
Author(s):  
Douglas Fraser-Pitt ◽  
Derry Mercer ◽  
Emma Lovie ◽  
Jennifer Robertson ◽  
Deborah O'Neil
Keyword(s):  
Cystic Fibrosis ◽  
Biofilm Formation ◽  
Burkholderia Cepacia ◽  
Standard Of Care ◽  
Burkholderia Cepacia Complex ◽  
Content Type ◽  
Folate Pathway ◽  
The Impact ◽  
Slow Growing

ABSTRACTThere are no wholly successful chemotherapeutic strategies againstBurkholderia cepaciacomplex (BCC) colonization in cystic fibrosis (CF). We assessed the impact of cysteamine (Lynovex) in combination with standard-of-care CF antibioticsin vitroagainst BCC CF isolates by the concentration at which 100% of bacteria were killed (MIC100) and checkerboard assays under CLSI standard conditions. Cysteamine facilitated the aminoglycoside-, fluoroquinolone- and folate pathway inhibitor-mediated killing of BCC organisms that were otherwise resistant or intermediately sensitive to these antibiotic classes. Slow-growing BCC strains are often recalcitrant to treatment and form biofilms. In assessing the impact of cysteamine on biofilms, we demonstrated inhibition of BCC biofilm formation at sub-MIC100s of cysteamine.

scholarly journals Pseudomonas aeruginosa-Derived Rhamnolipids and Other Detergents Modulate Colony Morphotype and Motility in the Burkholderia cepacia Complex

2017 ◽  
Vol 199 (13) ◽  
Author(s):  
Steve P. Bernier ◽  
Courtney Hum ◽  
Xiang Li ◽  
George A. O'Toole ◽  
Nathan A. Magarvey ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Burkholderia Cepacia ◽  
Burkholderia Cepacia Complex ◽  
Interspecies Interactions ◽  
Content Type ◽  
Low Concentrations ◽  
Subinhibitory Concentrations ◽  
The Impact ◽  
Polymicrobial Infections

ABSTRACT Competitive interactions mediated by released chemicals (e.g., toxins) are prominent in multispecies communities, but the effects of these chemicals at subinhibitory concentrations on susceptible bacteria are poorly understood. Although Pseudomonas aeruginosa and species of the Burkholderia cepacia complex (Bcc) can exist together as a coinfection in cystic fibrosis airways, P. aeruginosa toxins can kill Bcc species in vitro. Consequently, these bacteria become an ideal in vitro model system to study the impact of sublethal levels of toxins on the biology of typical susceptible bacteria, such as the Bcc, when exposed to P. aeruginosa toxins. Using P. aeruginosa spent medium as a source of toxins, we showed that a small window of subinhibitory concentrations modulated the colony morphotype and swarming motility of some but not all tested Bcc strains, for which rhamnolipids were identified as the active molecule. Using a random transposon mutagenesis approach, we identified several genes required by the Bcc to respond to low concentrations of rhamnolipids and consequently affect the ability of this microbe to change its morphotype and swarm over surfaces. Among those genes identified were those coding for type IVb-Tad pili, which are often required for virulence in various bacterial pathogens. Our study demonstrates that manipulating chemical gradients in vitro can lead to the identification of bacterial behaviors relevant to polymicrobial infections. IMPORTANCE Interspecies interactions can have profound effects on the development and outcomes of polymicrobial infections. Consequently, improving the molecular understanding of these interactions could provide us with new insights on the possible long-term consequences of these chronic infections. In this study, we show that P. aeruginosa-derived rhamnolipids, which participate in Bcc killing at high concentrations, can also trigger biological responses in Burkholderia spp. at low concentrations. The modulation of potential virulence phenotypes in the Bcc by P. aeruginosa suggests that these interactions contribute to pathogenesis and disease severity in the context of polymicrobial infections.

scholarly journals Is Melioidosis a One Health-Neglected Disease in Iran?

2020 ◽  
Vol 8 (3) ◽  
pp. 93-95
Author(s):  
Zahra Baradaran-Seyed
Keyword(s):  
Burkholderia Cepacia ◽  
Burkholderia Pseudomallei ◽  
Soil Structure ◽  
One Health ◽  
Health Surveillance ◽  
Burkholderia Cepacia Complex ◽  
Burkholderia Mallei ◽  
Isolation And Identification ◽  
History Of ◽  
The One

According to the several scientific resources, Iran is considered to be among the melioidosis-endemic regions of the world; this is in stark contrast to the domestic stance in Iran, where the risk of melioidosis is speculated only as an emerging infectious disease in a non-endemic area. Recently, we have had devastating flash floods in the most provinces of Iran; the changing of soil structure may facilitate the cultivation of bacteria in the upper layers of the ground, consequently causes exposure and re-emergence of the disease. Considering previous isolation and identification of Burkholderia mallei, Burkholderia Pseudomallei, and Burkholderia cepacia complex from human, animal, and environment samples in Iran, it is urgently necessary to upgrade and equip infrastructures to identify and distinguish between pathogenic Burkholderia and commensals in the context of the One Health surveillance system. In the present perspective, the history of melioidosis in the country is presented to attract attention to the negligence of disease over the last four decades.

scholarly journals Cost-effectiveness of rapid laboratory-based mass-spectrometry diagnosis of bloodstream infection: evidence from the RAPIDO randomised controlled trial

BMJ Open ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. e044623
Author(s):  
Padraig Dixon ◽  
William Hollingworth ◽  
Katie Pike ◽  
Rosy Reynolds ◽  
Margaret Stoddart ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cost Effectiveness ◽  
Bloodstream Infection ◽  
Controlled Trial ◽  
Rapid Identification ◽  
Clinical Event ◽  
Primary Analysis ◽  
Microbial Identification ◽  
Maldi Tof ◽  
The Impact

Objectives and interventionBloodstream infection, the presence of viable micro-organisms in the blood, is a prevalent clinical event associated with substantial mortality. Patient outcomes may be improved when the causative micro-organism is identified quickly. We assessed the cost-effectiveness of rapid microbial identification by matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometry.DesignEconomic evaluation alongside a randomised multicentre trial (RAPIDO: RAPId Diagnosis on Outcome) assessing the impact of rapid identification by MALDI-TOF spectrometry.SettingAdult inpatients with bloodstream infections at seven National Health Service hospital trusts in England and Wales.Primary outcomeNet monetary benefit, estimated as incremental costs compared with incremental 28-day survival, of rapid identification by MALDI-TOF spectrometry compared with conventional identification.MethodsPatients were randomised (1:1) to receive diagnosis by conventional methods of microbial identification (conventional arm) only or by MALDI-TOF spectrometry in addition to conventional identification (RAPIDO arm).ResultsData from 5550 patients were included in primary analysis. Mean imputed costs in 2018/2019 prices per patient were lower by £126 in the RAPIDO arm (95% CI −£784 to £532) but the proportion of patients alive at day 28 was lower (81.4% vs 82.3%). The probability of cost-effectiveness of MALDI-TOF was <0.5 at cost-effectiveness thresholds between £20 000 and £50 000.ConclusionsAdjunctive MALDI-TOF diagnosis was unlikely to be cost-effective when measured as cost per death avoided at 28 days. However, the differences between arms in cost and effect were modest, associated with uncertainty and may not accurately reflect ‘real-world’ routine use of MALDI-TOF technology in this patient group.Trial registration numbersISRCTN97107018/UKCRN 11978.

scholarly journals Discovery of a Novel Lineage Burkholderia cepacia ST 1870 Endophytically Isolated from Medicinal Polygala paniculata Which Shows Potent In Vitro Antileishmanial and Antimicrobial Effects

2021 ◽  
Vol 2021 ◽  
pp. 1-17
Author(s):  
Felipe de Paula Nogueira Cruz ◽  
Ailton Ferreira de Paula ◽  
Camila Tita Nogueira ◽  
Paulo Henrique Marques de Andrade ◽  
Leonardo Maurici Borges ◽  
...  
Keyword(s):  
Burkholderia Cepacia ◽  
Plant Pathogens ◽  
Leishmania Major ◽  
Micrococcus Luteus ◽  
Neglected Diseases ◽  
Isolation And Identification ◽  
Moniliophthora Perniciosa ◽  
Fungal Plant Pathogens ◽  
A Genome

In this study, we report the isolation and identification of an endophytic strain of Burkholderia cepacia (COPS strain) associated with Polygala paniculata roots. Polygala plants are rich sources of promising microbiomes, of which the literature reports several pharmacological effects, such as trypanocidal, antinociceptive, anesthetic, anxiolytics, and anticonvulsant activities. B. cepacia COPS belongs to a new sequence type (ST 1870) and harbors a genome estimated in 8.3 Mbp which exhibits the aminoglycosides and beta-lactams resistance genes aph(3′)-IIa and blaTEM-116, respectively. Analysis performed using MLST, average nucleotide identity, and digital DNA-DNA hybridization support its species-level identification and reveals its novel housekeeping genes alleles gyrB, lepA, and phaC. The root endophyte B. cepacia COPS drew our attention from a group of 14 bacterial isolates during the primary screening for being potentially active against Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212, Micrococcus luteus ATCC 9341, Escherichia coli ATCC 25922, and Candida albicans ATCC 10231 and exhibited the broad-spectrum activity against phytopathogenic fungi. In addition, COPS strain showed production of protease, lipase, and esterase in solid media, and its natural product extract showed potent inhibition against fungal plant pathogens, such as Moniliophthora perniciosa, whose antagonism index (89.32%) exceeded the positive control (74.17%), whereas Sclerotinia sclerotiorum and Ceratocystis paradoxa showed high percentages of inhibition (85.53% and 82.69%, respectively). COPS crude extract also significantly inhibited S. epidermidis ATCC 35984, E. faecium ATCC 700221 (MIC values of 32 μg/mL for both), E. faecalis ATCC 29212 (64 μg/mL), and S. aureus ATCC 25923 (128 μg/mL). We observed moderate antagonistic activity against A. baumannii ATCC 19606 and E. coli ATCC 25922 (both at 512 μg/mL), as well as potent cytotoxic effects on Leishmania infantum and Leishmania major promastigote forms with 78.25% and 57.30% inhibition. In conclusion, this study presents for the first time the isolation of an endophytic B. cepacia strain associated with P. paniculata and enough evidence that these plants may be considered a rich source of microbes for the fight against neglected diseases.

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