Achromobacter Spp Research Articles

Efficacy of Achromobacter Spp. Isolated from Rhizosphere of Soybean Against Macrophomina phaseolina

Soybean is an important oil seed crop produced in India, Madhya Pradesh is the largest soybean producing state. Charcoal rot caused by Macrophomina phaseolina is a major threat to soybean production leading to 30-50% yield loss. Biological control is an effective measure for management of this disease. Soil rhizosphere is a rich source of beneficial microbes able to suppress plant pathogens and promote plant growth. Present study aimed at isolating and characterizing indigenous antagonistic bacteria against M. phaseolina causing charcoal rot in soybean.

Mobilome Analysis of Achromobacter spp. Isolates from Chronic and Occasional Lung Infection in Cystic Fibrosis Patients.

Achromobacter spp. is an opportunistic pathogen that can cause lung infections in patients with cystic fibrosis (CF). Although a variety of mobile genetic elements (MGEs) carrying antimicrobial resistance genes have been identified in clinical isolates, little is known about the contribution of Achromobacter spp. mobilome to its pathogenicity. To provide new insights, we performed bioinformatic analyses of 54 whole genome sequences and investigated the presence of phages, insertion sequences (ISs), and integrative and conjugative elements (ICEs). Most of the detected phages were previously described in other pathogens and carried type II toxin-antitoxin systems as well as other pathogenic genes. Interestingly, the partial sequence of phage Bcep176 was found in all the analyzed Achromobacter xylosoxidans genome sequences, suggesting the integration of this phage in an ancestor strain. A wide variety of IS was also identified either inside of or in proximity to pathogenicity islands. Finally, ICEs carrying pathogenic genes were found to be widespread among our isolates and seemed to be involved in transfer events within the CF lung. These results highlight the contribution of MGEs to the pathogenicity of Achromobacter species, their potential to become antimicrobial targets, and the need for further studies to better elucidate their clinical impact.

Description of a novel selective medium (abscessus selective agar; asa) for the isolation of mycobacterium abscessus complex

Aims and objectives: Mycobacterium abscessus complex consisting of M. abscessus subsp. massiliense, M. abscessus subsp. bolletii and M. abscessus subsp. abscessus, have now emerged as clinically important NTM pathogens in patients with cystic fibrosis (CF). Culturally, these organisms are difficult to isolate from sputum amongst mixed populations of extremely rapidly growing Gram-negatives, particularly Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Achromobacter spp. Our objective was to develop a highly selective medium, not requiring decontamination of CF sputum but which exploited M. abscessus complex's resistance to several antibiotics. Methods: ASA medium was formulated employing Glucose, 16g/l; Agar, 20g/l; Yeast extract, 30g/l; Peptone, 6.8g/l; Chloramphenicol, 50mg/l; Ceftazidime, 32mg/l; Colistin, 24mg/l; Trimethoprim, 21.33mg/l; Sulfamethoxazole, 106.67mg/l and Novobiocin, 50mg/l. Mycobacterium abscessus complex organisms consisting of M. abscessus subsp. massiliense (n=2), M. abscessus subsp. bolletii (n=2) and M. abscessus subsp. abscessus (n=2) [circa 106 colony forming units (CFU)] were plated onto ASA medium and cultured aerobically for 2 weeks at 30o C. In addition, the following non-NTM organisms were also plated onto ASA medium, as above, Gram-positive: Enterococcus faecalis, Staphylococcus aureus (n=6), Staphylococcus aureus (mucoid), MRSA (Hospital-associated), MRSA (Livestock-associated cc398); Gram-negative: Achromobacter sp. (n=4), Burkholderia gladioli, Burkholderia multivorans, Escherichia coli (n=2), Pseudomonas aeruginosa (n=7), Salmonella enterica subsp. enterica serotype Nottingham, Salmonella enterica subsp. enterica serotype Typhimurium & Stenotrophomonas maltophilia. Following incubation, growth was assessed semiquantitatively, as +++, ++, +, few or no growth. Results: ASA medium supported the prolific growth (+++) of all M. abscessus complex organisms, but inhibited all non NTM organisms from growing (no growth). Conclusions: ASA medium is highly selective, which allowed for the proliferation of all members of the Mycobacterium abscessus complex, but successfully inhibited all other bacterial flora in CF sputum. This medium shows potential in isolating M. abscessus complex organisms from polymicrobial specimens/environments.

1620. Minocycline Activity Against Unusual Clinically Significant Gram-Negative Pathogens

BackgroundUnusual non-glucose fermenting Gram-negative (NFGN) pathogens, including Burkholderia cepacia species complex, Achromobacter spp, Alcaligenes spp, Aeromonas spp, and other genera, can cause serious hospital-acquired infections in immunocompromised patients. Some genera are inherently resistant to common drug classes and can acquire other resistance mechanisms, making them difficult to treat. In this study, we analyzed the susceptibility of NFGN isolates to minocycline (MIN). Isolates were collected as part of the SENTRY Antimicrobial Surveillance Program from 2014-2019.MethodsFrom 2014-2019, unusual NFGN isolates were collected from hospitalized patients in 102 hospitals in 35 countries on 4 continents. Hospitals submitted 1 isolate per patient per infection episode that met local criteria for being the likely causative pathogen. Identification was performed by the submitting laboratory and confirmed by JMI Laboratories with matrix-assisted laser desorption ionization-time of flight mass spectrometry or other molecular methods as required. Isolates were tested for MIN susceptibility using the CLSI broth microdilution method at JMI Laboratories. All infection types were included in the susceptibility analysis.ResultsThe most common infection from which the NFGN were isolated was pneumonia. The top 5 NFGN species were Achromobacter xylosoxidans (n=202), Burkholderia cepacia species complex (n=199), unspeciated Achromobacter (n=190), Aeromonas spp (n=127), including Aeromonas hydrophila (n=35), Chryseobacterium spp (n=59), and Alcaligenes faecalis (n=42). The % susceptible and MIC50/90 values of MIN for these species are shown in the table.ConclusionMIN had > 85% susceptible for the most frequently isolated unusual NFGN, including 92% susceptible for Achromobacter spp. and 85.9% for B. cepacia. These data suggest that MIN remains a useful treatment option for infections caused by unusual NFGN.Activities of MIN when tested against NFGN isolates DisclosuresS. J. Ryan Arends, PhD, Allergan (Research Grant or Support)Cipla Ltd. (Research Grant or Support)GlaxoSmithKline (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support) Jennifer M. Streit, BS, A. Menarini Industrie Farmaceutiche Riunite S.R.L. (Research Grant or Support)A. Menarini Industrie Farmaceutiche Riunite S.R.L. (Research Grant or Support)Allergan (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Merck (Research Grant or Support)Paratek Pharma, LLC (Research Grant or Support) Mariana Castanheira, PhD, 1928 Diagnostics (Research Grant or Support)A. Menarini Industrie Farmaceutiche Riunite S.R.L. (Research Grant or Support)Allergan (Research Grant or Support)Allergan (Research Grant or Support)Amplyx Pharmaceuticals (Research Grant or Support)Cidara Therapeutics (Research Grant or Support)Cidara Therapeutics (Research Grant or Support)Cipla Ltd. (Research Grant or Support)Cipla Ltd. (Research Grant or Support)Fox Chase Chemical Diversity Center (Research Grant or Support)GlaxoSmithKline (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Merck (Research Grant or Support)Merck (Research Grant or Support)Merck & Co, Inc. (Research Grant or Support)Merck & Co, Inc. (Research Grant or Support)Paratek Pharma, LLC (Research Grant or Support)Pfizer (Research Grant or Support)Qpex Biopharma (Research Grant or Support) Robert K. Flamm, PhD, A. Menarini Industrie Farmaceutiche Riunite S.R.L. (Research Grant or Support)Amplyx Pharmaceuticals (Research Grant or Support)Basilea Pharmaceutica International, Ltd (Research Grant or Support)Department of Health and Human Services (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)

1694. Successful Gut Decolonization of Extended-Spectrum β-lactamase Producing Klebsiella pneumoniae Using Oral Lyophilized Fecal Microbiota Transplant (FMT) in a Woman with Recurrent Urinary Tract Infections

BackgroundIn patients with anatomic disruption or long-term indwelling catheters of the urinary tract, recurrent infections can be a problematic complication. Exposure to multiple antibiotics can set the stage for the acquisition of resistant organisms. Restoration of a healthy gut microbiota may help such patients develop colonization resistance and eliminate multi-drug resistant organisms. We report the successful use of an oral FMT to decolonize an ESBL-producing K. pneumoniae (Kpn) from a woman with an ileal conduit with urostomy and recurrent urinary tract infections (UTI).MethodsFMT was performed using PRIM-DJ2727, an oral encapsulated lyophilized stool product under investigation for treatment of Clostridioides difficile infection. Three doses of PRIM-DJ2727 (60 g total fecal matter lyophilized to 1g/dose) were given weekly under an Expanded Access Investigational New Drug Application protocol approved by the US Food and Drug Administration and the local Institutional Review Board. Urine and stool samples were collected prior to treatment, 1 week after the final FMT dose, at transplant day +70, and transplant day +180. Samples underwent nucleic acid extraction using the Qiagen DNeasy PowerSoli Kit and 16S rRNA sequencing on an Illumina MiSeq.ResultsA 50 year old woman with von Willebrand disease presented with recurrent UTIs after complications from a hysterectomy decades prior. She had an ileal conduit with urostomy, and for the prior 2 years had been colonized with an ESBL Kpn with recurrent episodes of pyelonephritis. In the preceding 6 months, she had 5 symptomatic UTIs (Fig 1). FMT was given 1 week after stopping antibiotics from the most recent UTI. In the 6 months subsequent to the FMT, she developed two symptomatic UTIs, with cultures positive for Achromobacter (Axyl), Stenotrophomonas (Steno), and Enterococcus spp. Axyl and Steno were not identified in the FMT product (Fig 2A). Stool α-diversity increased after the transplant, and recovered by 6 months despite oral fluoroquinolone therapy (Fig 2B). Interestingly, there was an inverse relationship between stool and urine α-diversity. ESBL Kpn was not recovered subsequent to the FMT.Figure 1 Figure 2 ConclusionOral FMT was used to successfully decolonize a woman with recurrent UTIs due to ESBL Kpn.Disclosures Zhi-Dong Jiang, MD, DrPH, UT School of Public Health (Other Financial or Material Support, Patent for PRIM-DJ2727) Herbert DuPont, MD, UT School of Public Health (Other Financial or Material Support, Patent for PRIM-DJ2727) Cesar A. Arias, MD, MSc, PhD, FIDSA, Entasis Therapeutics (Scientific Research Study Investigator)MeMed (Scientific Research Study Investigator)Merck (Grant/Research Support) William R. Miller, MD, Entasis Therapeutics (Scientific Research Study Investigator)Merck (Grant/Research Support)Shionogi (Advisor or Review Panel member)

COVID-19 in cystic fibrosis patients

Since the beginning of the COVID-19 epidemic, the European cystic fibrosis society (ECFS) has decided to launch a special ECFS-COVID-19 program to collect information on the of COVID-19 characteristics in the patients with cystic fibrosis (CF). The results of the program should help timely and efficiently provide the patients with CF with the necessary care. Initially, it was assumed that COVID-19 would be severe in CF patients. The aim. Тo assess the prevalence and characteristics of COVID-19 in patients with cystic fibrosis (CF) in the Russian Federation (RF). Methods. 6 cases (4 children and 2 adults) of COVID-19 in Russian CF patients were analyzed. Results. There are 405,843 infected with SARS-CoV-2 in Russia, the incidence of coronavirus infection in Russia was 1.4 cases per 1 thousand people. According to the Ministry of Health of the RF, as of December 2019, there were 3,931 patients with CF (2,823 children and 1,108 adults). The incidence of COVID-19 was 1.5 per 1000 patients with CF (1.4 : 1,000 for children and 1.8 : 1,000 for adults). The incidence was not higher than in the General population. The diagnosis of COVID-19 was confirmed in 4 boys and 2 women, 3 of the patients were infected with Pseudomonas aeruginosa and 2 – with Achromobacter spp. Mild disease was seen in 5 patients including all the children. Pneumonia was registered in 3 patients. One child with COVID-19 had abdominal syndrome. 2 patients – 1 adult and 1 child – needed in-patient care. Additional antibiotics were given to 4 patients, 2 of them received i/v antibiotics. One adult patient was on the lung transplantation waiting list. This woman had long-term oxygen therapy and BiPAP noninvasive respiratory support before the infection with SARS-CoV-2, FEV1 was 24 %pred. Conclusion. Despite the fact that patients with CF are at risk of severe COVID-19, to date, in the described cases, COVID-19 infection has not led to a significant deterioration of the symptoms of CF. Not a single fatal outcome in Russian patients with CF has been recorded.

Bronchoscope-Related Outbreaks and Pseudo-Outbreaks: CDC Consultations—United States, 2014–2019

Background: Exposure to medical devices can be a risk factor for the development of healthcare-associated infections; bronchoscopes are a leading cause of device-associated outbreaks. We describe bronchoscope-related outbreaks and pseudo-outbreaks reported to the Centers for Disease Control and Prevention’s Division of Healthcare Quality Promotion (DHQP), and we summarize investigation steps and control measures. Methods: We identified bronchoscope-related consultations with state and local health departments between July 1, 2014, and September 30, 2019, in the DHQP database. We abstracted data on patient symptoms, clinical culture results, investigation findings, and subsequent infection prevention and control interventions. Results: We identified 15 consultations involving 150 patients (range, 3–31 patients per consultation). Each consultation involved at least 1 cluster of the same organism. Organisms associated with bronchoscope-associated clusters were nontuberculous mycobacteria (n = 7), Candida spp (n = 3), Exophiala spp (n = 2), Pseudomonas aeruginosa (n = 2), Enterobacter spp (n = 2), and Raoultella planticola, Stenotrophomonas maltophilia, Achromobacter spp, Mycobacterium tuberculosis, and Aspergillus spp (1 each; 2 consultations involved multiple pathogens). Procedures from which these patient specimens were collected included bronchoalveolar lavage, bronchial wash, bronchial brushing, sputum swab, and lymph node biopsy. For the 7 outbreaks in which clinical data were available, 5 did not have patients with clinical infections related to the pathogen recovered. Two consultations involved pseudo-outbreaks: one involved contamination of specimen collection tubes and the other involved contamination of cultures within the laboratory. Potential underlying pathogen sources included contaminated bronchoscopes (inadequate reprocessing or device damage) (n = 10, 67%), use of nonsterile ice, water, or saline during the procedure (n = 4, 27%), contaminated specimen collection tubes (n = 1, 7%), contaminated bronchoscope suite (n = 1, 7%), and clinical laboratory contamination (n = 1, 7%). The most common interventions included improvement of reprocessing procedures (n = 5), removal of possibly damaged bronchoscopes (n = 4), and eliminating nonsterile ice and water exposures in bronchoscopy (n = 3). Conclusions: Water-related organisms were the most commonly identified pathogens in bronchoscope-related consultations, highlighting the important role that exposure to contaminated water during bronchoscopy and bronchoscope reprocessing might play in bronchoscopy-associated outbreaks and pseudo-outbreaks. During bronchoscope-related outbreaks identifying a common pathogen could indicate problems in bronchoscope handling or reprocessing, device damage, or exposure to nonsterile water.Funding: NoneDisclosures: None

A Cluster of Corneal Donor Rim Cultures Positive for Achromobacter Species Associated With Contaminated Eye Solution.

To investigate a cluster of corneoscleral rim cultures positive for Achromobacter species over a 6-month period at Massachusetts Eye and Ear. An increased rate of positive corneal donor rim cultures was noted at Massachusetts Eye and Ear between July and December 2017. Positive cultures were subjected to identification and antimicrobial susceptibility testing by phenotypic (MicroScan WalkAway) and genotypic (16S rDNA sequencing) methods. Samples of the eye wash solution (GeriCare) used in the eye bank were also evaluated. Antimicrobial activity of Optical-GS against Achromobacter spp. at 4°C and 37°C was assessed by time-kill kinetics assay. Of 99 donor rims cultured, 14 (14.1%) grew bacteria with 11 (78.6%) due to uncommon nonfermenting Gram-negative bacilli. These had been identified by standard automated methods as Achromobacter (n = 3), Alcaligenes (n = 3), Ralstonia (n = 2), Pseudomonas (n = 2), and Stenotrophomonas (n = 1). Eight of these 11 isolates were subsequently available for molecular identification, and all were identified as Achromobacter spp. Six bottles of eyewash solution were evaluated and were positive for abundant Achromobacter spp. (3.4 × 105 ± 1.1 CFU/mL). Optisol-GS had no bactericidal activity against Achromobacter spp. at 4°C after 24-hour incubation but was bactericidal at 37°C. None of the patients who had received the contaminated corneas developed postoperative infection. An eyewash solution arising from a single lot was implicated in the contamination of donor rims by Achromobacter spp. The isolates were able to survive in the Optisol-GS medium at the recommended storage temperature. This highlights the need to continue improving protocols for tissue preparation and storage.

Effect of temperature on microcystin-LR removal and lysis activity on Microcystis aeruginosa (cyanobacteria) by an indigenous bacterium belonging to the genus Achromobacter.

Microcystis is a frequent cyanobacterium bloom-forming with cosmopolitan distribution which can produce a hepatotoxin group called microcystins (MCs). These MCs are resistant to the traditional processes employed in the water treatment plants and they are often detected after conventional treatments. Because of this, the bio-removal studies have obtained a great interest in the last decades. In this work, a bacterial strain namely LG1 with the ability to remove microcystin-LR (MC-LR) under laboratory conditions was isolated from Rio de la Plata River and it was identified as Achromobacter spp. This ubiquitous bacterium was able to remove 79.5% MC-LR in 7days with average removal time of 3.33 ± 0.08, 3.06 ± 0.05, and 2.77 ± 0.05days at 28, 32, and 36 ± 1°C, being higher at high temperature (36°C) with an activation energy = 16.79 ± 1.99kJmol-1. LG1 grew better at higher temperature (from 28 to 36 ± 1°C) increasing the specific growth rate (μ) and reducing 2-fold the lag phase duration (LPD) without significant differences (p > 0.05) between maximum population density (MPD). In addition, LG1 showed a lysis activity on two M. aeruginosa native strains in 7days measured as chlorophyll a (Chl-a) concentration. The lysis activity increased around 2-fold when increasing the temperature from 28 to 36 ± 1°C. This is the first report of an indigenous bacterium belonging to the genus Achromobacter spp. isolated from the Rio de la Plata River with the capacity to remove MC-LR and lysis activity onM. aeruginosa.

Duplex real-time PCR assay for the simultaneous detection of Achromobacter xylosoxidans and Achromobacter spp.

Several members of the Gram-negative environmental bacterial genus Achromobacter are associated with serious infections, with Achromobacter xylosoxidans being the most common. Despite their pathogenic potential, little is understood about these intrinsically drug-resistant bacteria and their role in disease, leading to suboptimal diagnosis and management. Here, we performed comparative genomics for 158 Achromobacter spp. genomes to robustly identify species boundaries, reassign several incorrectly speciated taxa and identify genetic sequences specific for the genus Achromobacter and for A. xylosoxidans . Next, we developed a Black Hole Quencher probe-based duplex real-time PCR assay, Ac-Ax, for the rapid and simultaneous detection of Achromobacter spp. and A. xylosoxidans from both purified colonies and polymicrobial clinical specimens. Ac-Ax was tested on 119 isolates identified as Achromobacter spp. using phenotypic or genotypic methods. In comparison to these routine diagnostic methods, the duplex assay showed superior identification of Achromobacter spp. and A. xylosoxidans , with five Achromobacter isolates failing to amplify with Ac-Ax confirmed to be different genera according to 16S rRNA gene sequencing. Ac-Ax quantified both Achromobacter spp. and A. xylosoxidans down to ~110 genome equivalents and detected down to ~12 and ~1 genome equivalent(s), respectively. Extensive in silico analysis, and laboratory testing of 34 non- Achromobacter isolates and 38 adult cystic fibrosis sputa, confirmed duplex assay specificity and sensitivity. We demonstrate that the Ac-Ax duplex assay provides a robust, sensitive and cost-effective method for the simultaneous detection of all Achromobacter spp. and A. xylosoxidans and will facilitate the rapid and accurate diagnosis of this important group of pathogens.

Characterization of microbial species in the biodegradation of explosives, military shooting range, Kaduna, Nigeria

Kachia military firing range since 1965 in situ characterization of microbes present in explosives contaminated soils was investigated. Bacteria gram stain morphological and biochemical characterization of the different microbial isolates. Bacterial DNA extracted from soil samples was achieved using the 16SrRNA is amplified using Polymerase Chain Reaction with the following microbes (Lysini bacillus, Escherichia coli, Enterobacter spp, Bacillus subtilis, Klebsiella pneumomia, Achromobacter spp and Arcobacter spp) was confirmed and results compared with the sequence obtained from the nucleotide database of National Centre for Biotechnology Information (NCBI). Fungal species isolates are: Rhizopus spp, Aspergillus niger, Penicillium spp, Trametes versicolourand Phanorochate chrysoporium may adapted to metabolise the explosives and heavy metals contaminant xenobiotic by biodegradation. Percentage isolates occurrence: 75% Enterobacterspp (highest) and 33% Escherichia coli (lowest); 67% Aspergillus niger (highest); and 17% for Penicillium sppand Trametes versicolor (lowest) respectively. Microbial biodegradation of explosives is considered to be most favourable under co-metabolic conditions. Site study explosives treatment by bioremediation will requires bioaugmentation of isolated microbes for xenobiotic biodegradation. Explosives impacts on biodiversity was illuminated and treatments protocol

Needs assessment for novel Gram-negative antibiotics in US hospitals: a retrospective cohort study

BackgroundEvidence-based needs assessments for novel antibiotics against highly-resistant Gram-negative infections (GNIs) are scarce. We aimed to use real-world data from an electronic health record repository to identify treatment opportunities in US hospitals for GNIs resistant to all first-line drugs.MethodsFor this retrospective cohort study, population estimates with an unmet need for novel Gram-negative antibiotics were quantified using the Cerner Health Facts database (2009–15), aggregating episodes of infection in US hospitals with pathogens displaying difficult-to-treat resistance (DTR; resistance to carbapenems, other β-lactams, and fluoroquinolones) and episodes involving empirical coverage with reserve drugs (colistin or polymyxin B and aminoglycosides). Episodes displaying extended-spectrum cephalosporin resistance (ECR) were also estimated. Episodes were multiplied by site-specific and fixed 14-day treatment durations for conservative and liberal days-of-therapy (DOT) estimates and stratified by site and taxon. Hospital type-specific DOT rates were reliability adjusted to account for random variation; cluster analyses quantified contribution from outbreaks.FindingsAcross 2 996 271 inpatient encounters and 134 hospitals, there were 1352 DTR-GNI episodes, 1765 episodes involving empirical therapy with colistin or polymyxin B, and 16 632 episodes involving aminoglycosides. Collectively, these yielded 39·0 (conservative estimate) to 138·2 (liberal estimate) DOT per 10 000 encounters for a novel DTR-GNI-targeted drug, whereas greater treatment opportunities were identified for ECR (six times greater) and β-lactam susceptible GNIs (70 times greater). The most common DTR-GNI site and pathogen was lower respiratory (14·3 [43·3%] of 33 DOT per 10 000 encounters) and Pseudomonas aeruginosa (522 [38·1%] of 1371 episodes), whereas Enterobacteriaceae urinary-tract infections dominated the ECR or carbapenem-sparing niche (59·0% [5589 of 9535 episodes]) equating to 210·7 DOT per 10 000 encounters. DTR Stenotrophomonas maltophilia, Burkholderia spp, and Achromobacter spp represented less than 1 DOT per 10 000 encounters each. The estimated need for DTR-GNI-targeted antibiotics saw minor contributions by outbreaks and varied from 0·5 to 73·1 DOT per 10 000 encounters by hospital type.InterpretationSuspected or documented GNIs with no or suboptimal treatment options are relatively infrequent. Non-revenue-based strategies and innovative trial designs are probably essential to the development of antibiotics with improved effectiveness for these GNIs.FundingCenter for Drug Evaluation and Research, US Food and Drug Administration; Intramural Research Program, National Institutes of Health Clinical Center and the National Institute of Allergy and Infectious Diseases and the National Cancer Institute.

SCREENING OF BIOSURFACTANT PRODUCTION BY BACTERIAL STRAINS ISOLATED FROM {SUDHANSHU SHEKHAR, 2015 #18}OIL CONTAMINATED SITES NEAR GAS STATIONS IN EGYPT.

Biosurfactant producing bacteria (28 isolates) were isolated from 30 oil contaminated soils and tested for production of biosurfactant by different screening methods. About 75% of the isolated bacteria showed no blood hemolysis and their emulsification index for hexane and xylene ranges from 28-35% and 30-45% respectively. Biosurfactant producing ability was confirmed by other tests where about 40% of isolates showed positive oil spreading activity, hydrocarbon overlay and 67% showed to be positive for drop collapsing test. 16s RNA sequencing of the most active isolates revealed one Stenotrophomonas maltophila, two Bacillus spp. and two Achromobacter spp. Biosurfactants extracted from these isolates showed variable antimicrobial activity against Staphylococcus aureus, E. coli and Pseudomonas aeruginosa. This study demonstrated that the biosurfactants produced by these bacteria could be used in combination with antibiotics for treating bacterial infections. Further study is required for enhancing biosurfactant production by bacterial isolates to be used environmentally for bioremediation of oil contaminated soils.

Rhizobacteria producing ACC deaminase mitigate water-stress response in finger millet (Eleusine coracana (L.) Gaertn.).

The aim of the study was to examine the influence of single and consortia treatments of drought tolerant rhizobacteria producing ACC deaminase together with additional plant growth promoting (PGP) characteristics on finger millet growth, antioxidant and nutrient concentration under water-stressed and irrigated (no stress) conditions. These rhizobacteria belong to the Variovorax sp. Achromobacter spp. Pseudomonas spp. and Ochrobactrum sp. The single inoculant of RAA3 (Variovorax paradoxus) and a consortium inoculant of four bacteria, i.e., DPC9 (Ochrobactrum anthropi), DPB13 (Pseudomonas palleroniana), DPB15 (Pseudomonas fluorescens) and DPB16 (Pseudomonas palleroniana), significantly boosted the overall growth parameters and nutrient concentrations in leaves of finger millet. Moreover, elevated levels of the reactive oxygen species scavenging enzymes-superoxide dismutase (17.3%, 11.6%), guaiacol peroxidase (38.7%, 22.2%), catalase (33.7%, 21.3%) and ascorbate peroxidase (18.2%, 10.0%); cellular osmolytes-proline (41.5%, 25.0%), phenol (44.5%, 37.5%); higher leaf chlorophyll (64.4%, 30.8%) and a reduced level of hydrogen peroxide (50.7%, 59.5%) and malondialdehyde (48.4%,72.5%) were noted, respectively, after single inoculation of RAA3 and a consortium treatment by strains DPC9 + DPB13 + DPB15 + DPB16, in contrast with non-treated plants mainly under water-stressed conditions. This finding clearly illustrates that PGPB that express ACC deaminase along with additional PGP traits could be an efficient approach for improving plant health in environments, where agricultural practices are reliant on rain for water.

Сравнительная характеристика и особенности клинической картины заболевания взрослых больных муковисцидозом с хроническим инфицированием нижних дыхательных путей Pseudomonas aeruginosa и другой грамотрицательной неферментирующей флорой

Background: in Russia, the life expectancy of cystic fibrosis (CF) patients increased by 10 years in 2011–2017 being 55.49 years in 2017. However, the number of patients with the chronic infection caused by non-fermenting gram-negative bacilli (NFGNB), e.g., Burkholderia cepacia, Achromobacter spp. etc., increased as well. Aim: to evaluate the differences in the nutritional and functional status and the severity of mutations in CF patients with chronic Pseudomonas infection or NFGNB infection and to assess the sensitivity of P. aeruginosa to tobramycin in CF patients in the Northwest region of Russia. Patients and Methods: 31 patients with CF aged 18–43 years (18 men and 13 women) were examined. The duration of the study was 12 months. Spirometry, anthropometry, and sputum culture were performed. Results: P. aeruginosa alone was isolated in 18 patients (58%), Achromobacter spp. in 9 patients (29%), and Burkholderia spp. in 4 patients (13%). The patients were divided into two groups, i.e., patients with chronic Pseudomonas infection (group 1, n=18, 10 out of 18 patients with mucoid strains of P. aeruginosa) or chronic NFGNB infection (group 2, n=13). The median age and the mode age were 27 years and 27 years, respectively, in group 1 and 24 years and 22 years, respectively, in group 2. It was demonstrated that CF patients with chronic NFGNB infection are characterized by poorer nutritional status (p<0.05) but similar functional status and the severity of CFTR gene mutation compared to CF patients with chronic Pseudomonas infection. It was also shown that Р. aeruginosa is highly sensitive to tobramycin (94.4%). Conclusions: in CF patients, chronic lower respiratory tract infections with Burkholderia cepacia and Achromobacter spp. account for 41.9% of gram-negative rod infections. Further studies and drug sensitivity monitoring are needed. KEYWORDS: cystic fibrosis, DNA test, chronic infection with Pseudomonas aeruginosa, Burkholderia cepacia, Achromobacter spp, non-fermenting gram-negative bacilli, CFTR mutation, nutritional status, pulmonary function tests, inhaled antibiotic therapy. FOR CITATION: Makhmutova V.R., Gembitskaya T.E., Chermenskiy A.G. et al. Comparative characteristics and clinical presentation of cystic fibrosis in adults with chronic lower respiratory tract infections with Pseudomonas aeruginosa and other non-fermenting gram-negative bacilli. Russian Medical Inquiry. 2020;4(4):186–191. DOI: 10.32364/2587-6821-2020-4-4-186-191.

Achromobacter spp. healthcare associated infections in the French West Indies: a longitudinal study from 2006 to 2016

BackgroundBacteria of the Achromobacter genus, more particularly xylosoxidans species, are responsible for various healthcare associated infections (HAI) which are increasingly described since the last decade. Cystic fibrosis (CF) patients are considered as potential reservoirs in hospitals. We performed a retrospective study to estimate the frequencies of Achromobacter spp. HAI among patients from French West Indies, to determine characteristics of infected patients and establish a possible link between CF and infections.MethodsAll adults with at least one Achromobacter spp. positive sample and infection criteria in accordance with European official definitions of HAI, hospitalized in University Hospital of Martinique from 2006 to 2016 for more than 48 h, were included. Patient clinical features, immune status and underlying diseases were obtained from medical files. A list of CF patients was given by clinicians.Antibiotic-susceptibility profiles of the strains were determined using an automated method.ResultsMean incidence density was 0.038/1000 days of hospitalization. Achromobacter spp. HAI evolved as an endemic situation with a low but pretty much stable incidence rate over the 11-year observation period. An epidemic peak was noticed in 2013. Among the 66 included patients, 56.1% were immunocompetent and no one had CF. Pneumonia and bacteraemia were the two main HAI. Among the 79 isolated strains, 92.4% were resistant to at least 1 major antibiotic and 16.4% met the definition of multidrug-resistant bacteria.ConclusionsThis microorganism, little known in our country because of the scarcity of CF patients, represents a threat for both immunosuppressed and immunocompetent patients and a therapeutic challenge because of its high resistance.

Features of the treatment of the cystic fibrosis patient with mixed microbial respiratory infection, including Pandoraea Pnomenusa

Respiratory tract infection is a major cause of complications and death in patients with cystic fibrosis (CF). Transmissible strains of Gram-negative non-fermenting Burkholderiales bacteria: Burkholderia cepacia complex, Achromobacter spp, Pandoraea spp. are quite alarming. In the paperfirstly there is presented a clinical case with mixed microbial infection of the CF patient with involvement of Pandoraea pnomenusa. There are reportedfeatures of diagnosis and treatment of a patient throughout 20 years, described the data of the examination of the patient and his airway microbiota. Timely identification of P. pnomenusa with the use of mass spectrometry MALDI-TOF and the molecular genetic techniques was shown to contribute to the isolation of the patient in the hospital, his transfer to the inpatient substitution therapy, that prevented the cross-infection of other CF patients. Permanent microbiological control revealed an increase of the antibiotic resistance of P. pnomenusa. The congruence of the antibiotic therapy with resistome of all identified microorganisms has allowed to develop an effective treatment schedule, which promoted to the elimination of P. aeruginosa and P. pnomenusa in the sputum of CF patients to minor amounts and preservation of this balance of microorganisms in the next six months.

English

Production of polyhydroxyalkanoates (PHA) by bacteria isolated from spent engine oil (SEO) contaminated soils was investigated using nitrogen limitation in the production medium. Out of ten isolates, three were selected as the best based on their ability to metabolize SEO effectively and fluoresce orange when stained with Nile Blue A dye. Fourier Transform Infrared spectrophometer was used as a confirmatory test for PHA detection by tracking the carbonyl bands on the spectra. Optimum growth of the isolates occurred at 1% (v/v) SEO, pH 7, 37°C for K1+VE and Kar5+VE1 while for Kar5+VE2 was at 1% (v/v) SEO, PH 7, 40°C and 150 RPM. Sequencing of 16S rDNA partial genes grouped the isolates into 6 different genera: Ochrobactrum, Pseudoxanthomonas, Bodetella, Achromobacter, Alcaligenes and Acinetobacter species. Isolate K1+VE, identified as an Ochrobactrum produced Poly (3-hydroxyheptanoate) 46% (w/w), isolate Kar5+VE1, identified as Achromobacter spp. produced poly (3-hydroxybutyrate) 20% (w/w)) while isolate Kar5+VE2, identified as an Alcaligenes spp. produced a co-polymer poly-3- (3-hydroxybutyrate-Co-3-hydroxyoctanoate) 45% (w/w). Orthoxylene and ethylbenzene were the major hydrocarbons in spent engine oil before degradation while ethylhexanol was the major degradation product as identified by Gas Chromatography-Mass Spectrophotometry. The isolates were able to degrade hydrocarbons as well as produce polyhydroxyalkanoates. Key words: Polyhydroxyalkanoates, spent engine oil, 16S rDNA, co-polymer.

Evaluation of ACC-deaminase-producing rhizobacteria to alleviate water-stress impacts in wheat (Triticum aestivum L.) plants.

Application of plant-growth-promoting rhizobacteria (PGPR) is an environmentally sustainable option to reduce the effects of abiotic and biotic stresses on plant growth and productivity. Bacteria isolated from rain-fed agriculture field soils in the Central Himalaya Kumaun region, India, were evaluated for the production of 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase. Those producing ACC deaminase in high amounts were evaluated for their potential to improve wheat (Triticum aestivum L.) plant growth under irrigated and water-stress conditions in two glasshouse experiments. Some of the isolates also showed other plant-growth-promoting (PGP) traits, e.g., N2 fixation, siderophore production, and phosphate solubilization; however, strains with higher ACC deaminase activity showed the greatest effects. These were Variovorax paradoxus RAA3; Pseudomonas spp. DPC12, DPB13, DPB15, DPB16; Achromobacter spp. PSA7, PSB8; and Ochrobactrum anthropi DPC9. In both simulated irrigated and water-stress conditions, a single inoculation of RAA3 and a consortium of DPC9 + DPB13 + DPB15 + DPB16 significantly improved wheat plant growth and foliar nutrient concentrations and caused significant positive changes in antioxidant properties compared with noninoculated plants especially under water stress. These findings imply that PGPB having ACC deaminase activity together with other PGP traits could potentially be effective inoculants to improve the growth of wheat plants in water-stressed rain-fed environments.

Effect of respiratory Achromobacter spp. infection on pulmonary function in patients with cystic fibrosis.

Cystic fibrosis (CF) patients are susceptible to infection with Achromobacter spp., although its clinical significance remains controversial. The aim of this study was to investigate the clinical impact of infection with Achromobacter spp. in CF patients. CF outpatients with multiple sputum cultures and follow-up lung function tests were assigned to the case group (infected with Achromobacter spp.) or the control group (never infected with Achromobacter spp.) according to the isolation of Achromobacter spp. The Achromobacter spp. group included two subgroups, taking into consideration whether the isolation of Achromobacter spp. was intermittent or chronic. Baseline lung function tests and longitudinal behaviour were examined in relation to Achromobacter spp. status. A total of 190 CF patients were treated from January 2003 to December 2015 in the CF unit and 21 (11 %) had at least one positive culture for Achromobacter spp. Of these, 11/21 (52.4 %) patients were chronically infected with Achromobacter spp. An analysis of changes during follow-up showed the annual rate of FEV1 decline: -2.3±1.6 % in the Achromobacter spp. group compared to -1.1±0.9 % (P=0.02) in the control group. The chronically infected group also had a significantly greater decline in FEV1 compared to the control group (-2.9±1.9 vs -1.1±0.9; P=0.04). The mean number of annual pulmonary exacerbations during the study period was significantly higher in the case group (1.9±0.9 vs 1.1±0.8; P=0.03). The Achromobacter spp. status in CF shows a trend towards more severe airflow obstruction and an association with accelerated decline in lung function parameters.