Collection Of Strains Research Articles

The distribution of fitness effects of plasmid pOXA-48 in clinical enterobacteria.

Antimicrobial resistance (AMR) in bacteria is a major public health problem. The main route for AMR acquisition in clinically important bacteria is the horizontal transfer of plasmids carrying resistance genes. AMR plasmids allow bacteria to survive antibiotics, but they also entail physiological alterations in the host cell. Multiple studies over the last few years have indicated that these alterations can translate into a fitness cost when antibiotics are absent. However, due to technical limitations, most of these studies are based on analysing new associations between plasmids and bacteria generated in vitro, and we know very little about the effects of plasmids in their native bacterial hosts. In this study, we used a CRISPR-Cas9-tool to selectively cure plasmids from clinical enterobacteria to overcome this limitation. Using this approach, we were able to study the fitness effects of the carbapenem resistance plasmid pOXA-48 in 35 pOXA-48-carrying isolates recovered from hospitalized patients. Our results revealed that pOXA-48 produces variable effects across the collection of wild-type enterobacterial strains naturally carrying the plasmid, ranging from fitness costs to fitness benefits. Importantly, the plasmid was only associated with a significant fitness reduction in four out of 35 clones, and produced no significant changes in fitness in the great majority of isolates. Our results suggest that plasmids produce neutral fitness effects in most native bacterial hosts, helping to explain the great prevalence of plasmids in natural microbial communities.

Action of Ethanolic Extract of Holerrhena pubescens on Bacterial Strains

The need to screen efficient, safe, affordable, and readily available antimicrobial agents from various medicinal plants, for their potential antimicrobial activity has become critical in light of the rapid development of microbial resistance against chemotherapeutic drugs (mainly antibiotics). Despite notable advances in the biological and phytochemical evaluation in recent years, comprehensive reviews of Holarrhena pubescens are limited. The main objective of this study is to evaluate the antimicrobial activities of this plant extract. One significant use of plant extracts can be in treating bacterial infections and develop potential antibacterial agents against multi drug resistant (MDR) and susceptible bacteria. If antimicrobial activities are present, these extracts will provide useful information for discovering new compounds with better activity and more effectiveness against resistant and susceptible bacteria responsible for infections than currently available. The ethanolic extract of leaves of Holarrhena pubescens was prepared and tested against multidrug resistant (MDR) and American type culture collection strains of bacteria. We have obtained two types of results in this experiment. The extract showed antimicrobial properties against Staphylococcus aureus (ATCC 23235), Escherichia coli (MDR) and Acinetobacter (MDR), while, it showed no antimicrobial activities along with some growth promoting role against Escherichia coli (ATCC 25922), and Klebsialla pneumoniae (MDR), which may be due to chemicals other than secondary metabolites.

Uncovering Genomic Features and Biosynthetic Gene Clusters in Endophytic Bacteria from Roots of the Medicinal Plant Alkanna tinctoria Tausch as a Strategy To Identify Novel Biocontrol Bacteria.

The world's population is increasing at a rate not seen in the past. Agriculture, providing food for this increasing population, is reaching its boundaries of space and natural resources. In addition, changing legislation and increased ecological awareness are forcing agriculture to reduce its environmental impact. This entails the replacement of agrochemicals with nature-based solutions. In this regard, the search for effective biocontrol agents that protect crops from pathogens is in the spotlight. In this study, we have investigated the biocontrol activity of endophytic bacteria isolated from the medicinal plant Alkanna tinctoria Tausch. To do so, an extensive collection of bacterial strains was initially genome sequenced and in silico screened for features related to plant stimulation and biocontrol. Based on this information, a selection of bacteria was tested in vitro for antifungal activity using direct antagonism in a plate assay and in planta with a detached-leaf assay. Bacterial strains were tested individually and in combinations to assess the best-performing treatments. The results revealed that many bacteria could produce metabolites that efficiently inhibit the proliferation of several fungi, especially Fusarium graminearum. Among these, Pseudomonas sp. strain R-71838 showed a strong antifungal effect, in both dual-culture and in planta assays, making it the most promising candidate for biocontrol application. Using microbes from medicinal plants, this study highlights the opportunities of using genomic information to speed up the screening of a taxonomically diverse set of bacteria with biocontrol properties. IMPORTANCE Phytopathogenic fungi are a major threat to global food production. The most common management practice to prevent plant infections involves the intensive use of fungicides. However, with the growing awareness of the ecological and human impacts of chemicals, there is a need for alternative strategies, such as the use of bacterial biocontrol agents. Limitations in the design of bacterial biocontrol included the need for labor-intensive and time-consuming experiments to test a wide diversity of strains and the lack of reproducibility of their activity against pathogens. Here, we show that genomic information is an effective tool to select bacteria of interest quickly. Also, we highlight that the strain Pseudomonas sp. R-71838 produced a reproducible antifungal effect both in vitro and in planta. These findings build a foundation for designing a biocontrol strategy based on Pseudomonas sp. R-71838.

Antifungal Properties of Ethanolic Extract of Holarrhena Pubescens against Candida Species

Antimicrobial resistance is a global problem that jeopardizes the efficient use of modern medicine and the state of the world's health. The effectiveness of traditional antifungals used to treat fungal infections has declined, increasing mortality. Following exposure to antifungal medications, acquired mechanisms can lead to the development of resistance to currently available antifungal medicines. Invasive fungal infections have few treatment choices, and patients who are most at risk frequently have other co-morbid conditions, such as immunosuppression. This problem is made worse by the paucity of novel antifungals now under research. One potential research avenue for tackling the problem of MDR fungal pathogens is plants that are used in traditional medicine. When used to treat infectious diseases, antimicrobials derived from plants have a great deal of therapeutic promise and have less adverse effects than their synthetic counterparts. In this study, the possible antifungal action of ethanolic extract of Holarrhena pubescens was tested against American Type Culture Collection (ATCC) and Multidrug resistant (MDR) strains of pathogenic fungi, Candida species. Antifungal activities of ethanolic extract of Holarrhena pubescens were studied by determining the minimum inhibitory concentration (MIC) value which was done by serial dilution in Mueller Hinton broth.The ethanolic extract of Holarrhena pubescensshowedantifungal activities exhibiting MIC values varying between 1.5625-3.125 mg/mL in both ATCC and MDR fungal pathogenic strains of Candida. Ethanolic extract of Holarrhena pubescenswas found to have effective antifungal activity against Candida species andcould be the cure to invasive candidiasis which is a global health threat.

Founding of the culture collection of antibiotic-resistant strains of zoonotic bacteria in the Russian Federation.

The main purpose of a national bioresource center is to standardize, centralize, preserve, and ensure accessibility of microbial bioresources that accumulate there because of state research programs. The establishment of national bioresource centers for antibiotic-resistant microorganisms allows to solve practical problems in the field of veterinary service, as well as to develop effective chemotherapeutic and disinfectant drugs to overcome the mechanisms of resistance. This study aimed to outline the process of forming a national culture collection of antibiotic-resistant strains of zoonotic bacteria in the Russian Federation using two microbial strains. The object of research was isolates of Salmonella spp., Escherichia coli, Enterococcus spp., Campylobacter spp., Listeria monocytogenes, and Staphylococcus spp., all of which were obtained from biomaterials of farm animals, feed samples, bedding, water from livestock buildings, washouts from environmental objects, and food products. The resistance of bacterial isolates was determined using microbiological and molecular-genetic research methods. During monitoring studies, 1489 bacterial isolates were isolated. In total, 408 bacterial isolates were tested for sensitivity to antimicrobial agents, including E. coli (47.6%), Salmonella spp. (30.4%), Enterococcus spp. (11.3%), and Campylobacter spp. (10.8%). For genetic characterization, 95 isolates of Salmonella enterica, E. coli, Campylobacter spp., L. monocytogenes, Staphylococcus spp., Enterococcus spp. were chosen from the research collection, which was formed as part of the monitoring program for antibiotic resistance. Deposited isolates that underwent whole-genome analysis can be used as positive control samples both in the development and use of methods or test systems for the detection of various resistance genes in zoonotic bacteria. In addition, such isolates can also be used for microbiological studies related to determining the sensitivity of microorganisms to antibacterial drugs, for phenotypic studies in the diagnosis of various bacterial infections in animals and birds, and retrospective analysis of strains from numerous collections.

Investigation of Delafloxacin Resistance in Multidrug-Resistant Escherichia coli Strains and the Detection of E. coli ST43 International High-Risk Clone.

Delafloxacin is a novel fluoroquinolone agent that is approved for clinical application. In this study, we analyzed the antibacterial efficacy of delafloxacin in a collection of 47 Escherichia coli strains. Antimicrobial susceptibility testing was performed by the broth microdilution method and minimum inhibitory concentration (MIC) values were determined for delafloxacin, ciprofloxacin, levofloxacin, moxifloxacin, ceftazidime, cefotaxime, and imipenem. Two multidrug-resistant E. coli strains, which exhibited delafloxacin and ciprofloxacin resistance as well as extended-spectrum beta-lactamase (ESBL) phenotype, were selected for whole-genome sequencing (WGS). In our study, delafloxacin and ciprofloxacin resistance rates were 47% (22/47) and 51% (24/47), respectively. In the strain collection, 46 E. coli were associated with ESBL production. The MIC50 value for delafloxacin was 0.125 mg/L, while all other fluoroquinolones had an MIC50 value of 0.25 mg/L in our collection. Delafloxacin susceptibility was detected in 20 ESBL positive and ciprofloxacin resistant E. coli strains; by contrast, E. coli strains that exhibited a ciprofloxacin MIC value above 1 mg/L were delafloxacin-resistant. WGS analysis on the two selected E. coli strains (920/1 and 951/2) demonstrated that delafloxacin resistance is mediated by multiple chromosomal mutations, namely, five mutations in E. coli 920/1 (gyrA S83L, D87N, parC S80I, E84V, and parE I529L) and four mutations in E. coli 951/2 (gyrA S83L, D87N, parC S80I, and E84V). Both strains carried an ESBL gene, blaCTX-M-1 in E. coli 920/1 and blaCTX-M-15 in E. coli 951/2. Based on multilocus sequence typing, both strains belong to the E. coli sequence type 43 (ST43). In this paper, we report a remarkable high rate (47%) of delafloxacin resistance among multidrug-resistant E. coli as well as the E. coli ST43 international high-risk clone in Hungary.

Antibacterial activity and phytochemical components of leaf extract of Calpurnia aurea

Local Ethiopians use Calpurnia aurea to treat skin infections. However, there is no adequate scientific confirmation. The aim of this study was to evaluate the antibacterial activities of the crude and the fractionated extracts of C. aurea leaves against different bacterial strains. The crude extract was made by maceration. The Soxhlet extraction method was used to obtain fractional extracts. The antibacterial activity against gram positive and gram negative American Type Culture Collection (ATCC) strains was performed using the agar diffusion technique. The minimum inhibitory concentration was determined through the microtiter broth dilution method. Preliminary phytochemical screening was done using standard techniques. The largest yield was obtained from ethanol fractional extract. Except for chloroform, which provided a relatively low yield when compared to petroleum ether, increasing the polarity of the extracting solvent improved the yield. The crude extract, solvent fractions, and the positive control showed inhibitory zone diameter, while the negative control did not. When used at a concentration of 75 mg/ml, the crude extract had similar antibacterial effects as gentamicin (0.1 mg/ml) and the ethanol fraction. The 2.5 mg/ml crude ethanol extract of C. aurea suppressed the growth of Pseudomonas aeruginosa, Streptococcus pneumoniae, and Staphylococcus aureus, according to the MIC values. The extract of C. aurea was more effective in inhibiting P. aeruginosa than the other gram-negative bacteria. Fractionation enhanced the antibacterial effect of the extract. All fractionated extracts showed the highest inhibition zone diameter against S. aureus. Petroleum ether extract had the greatest inhibition zone diameter against all bacterial strains. The non-polar components were more active compared to the more polar fractions. The phytochemical components discovered in the leaves of C. aurea included alkaloids, flavonoids, saponins, and tannins. Among these, the tannin content was remarkably high. The current results could provide a rational support for the traditional use of C. aurea to treat skin infections.

Artificial mixed microbial system for polycyclic aromatic hydrocarbons degradation.

Polycyclic aromatic hydrocarbons (PAHs) are environmental pollutants with major risks to human health. Biological degradation is environmentally friendly and the most appealing remediation method for a wide range of persistent pollutants. Meanwhile, due to the large microbial strain collection and multiple metabolic pathways, PAH degradation via an artificial mixed microbial system (MMS) has emerged and is regarded as a promising bioremediation approach. The artificial MMS construction by simplifying the community structure, clarifying the labor division, and streamlining the metabolic flux has shown tremendous efficiency. This review describes the construction principles, influencing factors, and enhancement strategies of artificial MMS for PAH degradation. In addition, we identify the challenges and future opportunities for the development of MMS toward new or upgraded high-performance applications.

Recombinant endopeptidases IdeS and IdeZ and their potential applications

IdeS and IdeZ endopeptidases are streptococcal virulence factors that specifically cleave IgG heavy chains and represent an interest from biotechnological, medical, and veterinary points of view. The genes encoding IdeS and IdeZ endopeptidases (the ideS gene was cloned from a Streptococcus pyogenes collection strain, and S. zooepidemicusideZ gene was synthesized) were cloned and expressed in Escherichia coli heterologous expression system, and 6His-tag affinity domain was introduced into the amino acid sequence of each endopeptidase. IdeS and IdeZ enzymes were isolated and purified by metal affinity chromatography. The resulting IdeS and IdeZ are apparently homogeneous on polyacrylamide gel electrophoresis and are active against human and animal IgG. The specificity of human IgG cleavage by endopeptidases IdeS and IdeZ was confirmed by polyacrylamide gel electrophoresis. Application of recombinant IdeZ was demonstrated for immunological analysis of equine “strangles” (diagnostics and specific antibodies’ titer determination in the blood). Thus, IdeZ can be used in veterinary medicine and sanitary microbiology for the diagnosis of infections caused by S. equi and S. zooepidemicus in addition to their applications in medicine and biotechnology.

Chemoenzymatic approach towards the synthesis of the antitumor and antileishmanial marine metabolite (+)-Harzialactone A via the stereoselective, biocatalyzed reduction of a prochiral ketone

As a rich source of biological active compounds, marine natural products have been increasingly screened as candidates for developing new drugs. Among the several marine products and metabolites, (+)-Harzialactone A has drawn considerable attention for its antitumor and antileishmanial activity. In this work a chemoenzymatic approach has been implemented for the preparation of the marine metabolite (+)-Harzialactone A. The synthesis involved a stereoselective, biocatalyzed reduction of the prochiral ketone 4-oxo-5-phenylpentanoic acid or the corresponding esters, all generated by chemical reactions. A collection of different promiscuous oxidoreductases (both wild-type and engineered) and diverse microorganism strains were investigated to mediate the bioconversions. After co-solvent and co-substrate investigation in order to enhance the bioreduction performance, T. molischiana in presence of NADES (choline hydrochloride-glucose) and ADH442 were identified as the most promising biocatalysts, allowing the obtainment of the (S)-enantiomer with excellent ee (97% to > 99% respectively) and good to excellent conversion (88% to 80% respectively). The successful attempt in this study provides a new chemoenzymatic approach for the synthesis of (+)-Harzialactone A.

Plants, Cells, Algae, and Cyanobacteria In Vitro and Cryobank Collections at the Institute of Plant Physiology, Russian Academy of Sciences-A Platform for Research and Production Center.

Ex situ collections of algae, cyanobacteria, and plant materials (cell cultures, hairy and adventitious root cultures, shoots, etc.) maintained in vitro or in liquid nitrogen (-196 °C, LN) are valuable sources of strains with unique ecological and biotechnological traits. Such collections play a vital role in bioresource conservation, science, and industry development but are rarely covered in publications. Here, we provide an overview of five genetic collections maintained at the Institute of Plant Physiology of the Russian Academy of Sciences (IPPRAS) since the 1950-1970s using in vitro and cryopreservation approaches. These collections represent different levels of plant organization, from individual cells (cell culture collection) to organs (hairy and adventitious root cultures, shoot apices) to in vitro plants. The total collection holdings comprise more than 430 strains of algae and cyanobacteria, over 200 potato clones, 117 cell cultures, and 50 strains of hairy and adventitious root cultures of medicinal and model plant species. The IPPRAS plant cryobank preserves in LN over 1000 specimens of in vitro cultures and seeds of wild and cultivated plants belonging to 457 species and 74 families. Several algae and plant cell culture strains have been adapted for cultivation in bioreactors from laboratory (5-20-L) to pilot (75-L) to semi-industrial (150-630-L) scale for the production of biomass with high nutritive or pharmacological value. Some of the strains with proven biological activities are currently used to produce cosmetics and food supplements. Here, we provide an overview of the current collections' composition and major activities, their use in research, biotechnology, and commercial application. We also highlight the most interesting studies performed with collection strains and discuss strategies for the collections' future development and exploitation in view of current trends in biotechnology and genetic resources conservation.

Evaluation of the In Vitro Antifungal Activity of Novel Arylsulfonamides against Candida spp.

The antifungal activity of molecules belonging to the arylsulfonamide chemotype has previously been demonstrated. Here, we screened arylsulfonamide-type compounds against a range of Candida spp. and further established the structure-activity relationship based on a "hit compound". A series of four sulfonamide-based compounds, N-(4-sulfamoylbenzyl) biphenyl-4-carboxamide (3), 2,2-diphenyl-N-(4-sulfamoylbenzyl) acetamide (4), N-(4-sulfamoylphenethyl) biphenyl-4-carboxamide (5) and 2,2-diphenyl-N-(4-sulfamoylphenethyl) acetamide (6), were tested against the American Type Culture Collection (ATCC) and clinical strains of C. albicans, C. parapsilosis and C. glabrata. Based on the fungistatic potential of prototype 3, a further subset of compounds, structurally related to hit compound 3, was synthesized and tested: two benzamides (10-11), the related amine 4-[[(4-4-((biphenyl-4-ylmethylamino)methyl) benzenesulfonamide (13) and the corresponding hydrochloride, 13.HCl. Both amine 13 and its hydrochloride salt had fungicidal effects against Candida glabrata strain 33 (MFC of 1.000 mg/mL). An indifferent effect was detected in the association of the compounds with amphotericin B and fluconazole. The cytotoxicity of the active compounds was also evaluated. This data could be useful to develop novel therapeutics for topical use against fungal infections.

From genotype to phenotype: computational approaches for inferring microbial traits relevant to the food industry.

When selecting microbial strains for the production of fermented foods, various microbial phenotypes need to be taken into account to achieve target product characteristics, such as biosafety, flavor, texture, and health-promoting effects. Through continuous advances in sequencing technologies, microbial whole-genome sequences of increasing quality can now be obtained both cheaper and faster, which increases the relevance of genome-based characterization of microbial phenotypes. Prediction of microbial phenotypes from genome sequences makes it possible to quickly screen large strain collections in silico to identify candidates with desirable traits. Several microbial phenotypes relevant to the production of fermented foods can be predicted using knowledge-based approaches, leveraging our existing understanding of the genetic and molecular mechanisms underlying those phenotypes. In the absence of this knowledge, data-driven approaches can be applied to estimate genotype-phenotype relationships based on large experimental datasets. Here, we review computational methods that implement knowledge- and data-driven approaches for phenotype prediction, as well as methods that combine elements from both approaches. Furthermore, we provide examples of how these methods have been applied in industrial biotechnology, with special focus on the fermented food industry.

Characterization of Silver Nanoparticle Systems from Microalgae Acclimated to Different CO2 Atmospheres.

Green synthesis of metallic nanoparticles using microalgae exposed to high CO2 atmospheres has not been studied in detail; this is of relevance in biological CO2 mitigation systems where considerable biomass is produced. In this study, we further characterized the potential of an environmental isolate Desmodesmus abundans acclimated to low and high CO2 atmospheres [low carbon acclimation (LCA) and high carbon acclimation (HCA) strains, respectively] as a platform for silver nanoparticle (AgNP) synthesis. As previously characterized, cell pellets at pH 11 were selected from the biological components tested of the different microalgae, which included the culture collection strain Spirulina platensis. AgNP characterization showed superior performance of strain HCA components as preserving the supernatant resulted in synthesis in all pH conditions. Size distribution analysis evidenced strain HCA cell pellet platform (pH 11) as the most homogeneous AgNP population (14.9 ± 6.4 nm diameter, -32.7 ± 5.3 mV) followed by S. platensis (18.3 ± 7.5 nm, -33.9 ± 2.4 mV). In contrast, strain LCA presented a broader population where the size was above 100 nm (127.8 ± 14.8 nm, -26.7 ± 2.4 mV). Fourier-transform infrared and Raman spectroscopies showed that the reducing power of microalgae might be attributed to functional groups in the cell pellet from proteins, carbohydrates, and fatty acids and, in the supernatant, from amino acids, monosaccharides, disaccharides, and polysaccharides. Microalgae AgNPs exhibited similar antimicrobial properties in the agar diffusion test against Escherichia coli. However, they were not effective against Gram (+) Lactobacillus plantarum. It is suggested that a high CO2 atmosphere potentiates components in the D. abundans strain HCA for nanotechnology applications.

Detection of bacteria in dental samples using the Periodic acid-Schiff (PAS) histological stain

The bacterial cell wall mainly consists of glycoproteins and polysaccharides, which could be detected in dental tissue with specific stain protocols. The present study aimed to investigate bacteria stainability in dental histological samples of human teeth by a histochemical method. Eight extracted teeth, because severely decayed, were decalcified, dehydrated, paraffin-embedded, and serially sectioned at 4 µm thickness each. The serial sections were then stained with Periodic acid-Schiff (PAS). Furthermore, SEM analysis was performed on the same slide of one previously histologically investigated tooth to acquire more details on the structures stained by the PAS method obtained from the histological procedures. Afterward, some American Type Culture Collection (ATCC) strains, smeared on glass slides, were stained following the staining method used in histological samples. Stained rod and cocci forms by PAS stain, observed under light microscopy, were predominantly detected inside dentinal tubules and root canal space of histologically examined specimens, suggesting their bacterial origin. Additional SEM analysis on the identical histological stained slide showed the precise nature of these forms (bacteria) and supplemental information regarding their vitality status. In addition, ATCC smeared strain samples showed variable PAS stainability of microorganisms investigated. Due to its properties, the PAS histochemical stain could be a valid and helpful aid for non- or weakly stainable microorganisms in infected tissues to be associated with other methods of investigation.

P251 Susceptibility to cefiderocol of an accurately identified collection of 110 Achromobacter strains from cystic fibrosis patients

Achromobacter spp. are emerging pathogens in CF. Based on intrinsic resistance and the report of multidrug resistant strains, Achromobacter spp. are considered as difficult-to-treat pathogens. Cefiderocol is a novel siderophore cephalosporin that has been infrequently used in the management of Achromobacter infections and for which susceptibility data are lacking. We evaluated the cefiderocol susceptibility of a collection of CF Achromobacter strains encompassing different airway colonization types, species and antimicrobial susceptibility patterns.

Recombinant Endopeptidases IdeS and IdeZ and Their Potential Application.

Endopeptidases IdeS and IdeZ (streptococcal virulence factors that specifically cleave IgG heavy chains) are of particular interest because of their potential use in biotechnology, medicine, and veterinary. Genes encoding these enzymes were cloned and expressed in Escherichia coli heterologous expression system (ideS was cloned from a Streptococcus pyogenes collection strain; ideZ from Streptococcus zooepidemicus was synthesized). The 6His-tag was introduced into the amino acid sequence of each endopeptidase, and IdeS and IdeZ were purified by metal affinity chromatography to an apparent homogeneity (according to polyacrylamide gel electrophoresis). Purified enzymes were active against human and animal IgGs; their specificity toward human IgGs was confirmed by polyacrylamide gel electrophoresis. Recombinant IdeZ was used for immunological analysis of equine strangles infection (diagnostics and determination of the titer of specific antibodies in blood). Hence, IdeZ can be used in veterinary and sanitary microbiology to diagnose infections caused by Streptococcus equi and S. zooepidemicus in addition to its application in medicine and biotechnology.

NanJ Is the Major Sialidase for Clostridium perfringens Type F Food Poisoning Strain 01E809.

Clostridium perfringens type F strains cause food poisoning (FP) when they sporulate and produce C. perfringens enterotoxin (CPE) in the intestines. Most type F FP strains carry a chromosomal cpe gene (c-cpe strains). C. perfringens produces up to three different sialidases, named NanH, NanI, and NanJ, but some c-cpe FP strains carry only nanJ and nanH genes. This study surveyed a collection of such strains and showed that they produce sialidase activity when cultured in Todd-Hewitt broth (TH) (vegetative cultures) or modified Duncan-Strong (MDS) medium (sporulating cultures). Sialidase null mutants were constructed in 01E809, a type F c-cpe FP strain carrying the nanJ and nanH genes. Characterization of those mutants identified NanJ as the major sialidase of 01E809 and showed that, in vegetative and sporulating cultures, nanH expression affects nanJ expression and vice versa; those regulatory effects may involve media-dependent changes in transcription of the codY or ccpA genes but not nanR. Additional characterization of these mutants demonstrated the following: (i) NanJ contributions to growth and vegetative cell survival are media dependent, with this sialidase increasing 01E809 growth in MDS but not TH; (ii) NanJ enhances 24-h vegetative cell viability in both TH and MDS cultures; and (iii) NanJ is important for 01E809 sporulation and, together with NanH, CPE production in MDS cultures. Lastly, NanJ was shown to increase CPE-induced cytotoxicity and CH-1 pore formation in Caco-2 cells. Collectively, these results suggest that NanJ may have a contributory role in FP caused by type F c-cpe strains that carry the nanH and nanJ genes.

VIRULENCE OF PSEUDOMONAS SYRINGAE STRAINS FROM SORIZ (SORGHUM ORYSOIDUM) AND THEIR SPECIALIZATION

Objective. To study the virulence properties of Pseudomonas syringae strains isolated from soriz and various representatives of sorghum crops in Ukraine and to establish the sensitivity range of plants to these pathogens. Methods. Ten strains freshly isolated from soriz (Sorghum orysoidum) were used to study virulence properties, namely: Pseudomonas syringae 210342, 211141a, 211141, 210341, 21034, 210541b, 210541, and Pseudomonas sp. 210521, 21052, 21054, pathogenic strains of sorghum crops from the collection of the Department of Phytopathogenic Bacteria of D. K. Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine and collection strains of phytopathogens that can be potential causative agents of bacterial diseases of soriz or are causative agents of diseases of agricultural crops that can be used in crop rotation. The virulence properties of strains isolated from sorghum were determined on plants of the genus Sorghum. To establish the range of crops sensitive to these pathogens, a number of plants belonging to other genera and families, both cultivated and weed, were tested. Artificial inoculation was carried out in the greenhouse and in the field as injection of bacterial suspension of 1×109 CFU/mL into the leaf surface and stem at all phases of the development of soriz and sorghum cultures. Accounting for artificial contamination was carried out in 7 to 14 days using a locally modified 6-point scale. Aggressiveness of bacteria was assessed by the size of necrosis that formed at the site of injection of the bacterial suspension. Aggressiveness was calculated as the arithmetic mean of the results. Results. P. syringae strains isolated from soriz were found to be virulent for soriz and sorghum crops, however, heterogeneous in aggressiveness. The most aggressive strains were P. syringae 211141a, 210341, and 21034. Some collection strains, isolated from different cultures, showed higher aggressiveness against sugar and grain sorghum than against soriz and Sudan grass. The studied strains of phytopathogens were found to affect segetal vegetation; the most sensitive to the studied strains were horsetail, field bindweed, goldenrod. They showed high aggressiveness against setaria, ragweed, and common sedge. Strains of P. syringae affecting potato, beet, sunflower and tomato plants were identified. Conclusion. Pseudomonas syringae strains isolated from contaminated sorghum plants are virulent and capable of infecting a wide range of plants, including plants of the sorghum family, other types of agricultural crops, and various types of segetal vegetation. Strains of Pectobacterium carotovorum, Xanthomonas vasicola pv. holcicola, Diskeya chryzanthemy, Pantoea agglomerans can be potential pathogens of sorghum crops. The results indicate that segetal vegetation can serve as a reservoir of bacterial infections and play a role in the transfer of bacterial pathogens.

Markers of epidemiological success of methicillin-resistant Staphylococcus aureus isolates in European populations

ObjectivesMethicillin-resistant Staphylococcus aureus (MRSA) infections impose a considerable burden on health systems, yet there is remarkable variation in the global incidence and epidemiology of MRSA. The MACOTRA consortium aimed to identify bacterial markers of epidemic success of MRSA isolates in Europe using a representative MRSA collection originating from France, the Netherlands and the United Kingdom. MethodsOperational definitions of success were defined in consortium meetings to compose a balanced strain collection of successful and sporadic MRSA isolates. Isolates were subjected to antimicrobial susceptibility testing and whole-genome sequencing; genes were identified and phylogenetic trees constructed. Markers of epidemiological success were identified using genome-based time-scaled haplotypic density analysis and linear regression. Antimicrobial usage data from ESAC-Net was compared with national MRSA incidence data. ResultsHeterogeneity of MRSA isolate collections across countries hampered the use of a unified operational definition of success; therefore, country-specific approaches were used to establish the MACOTRA strain collection. Phenotypic antimicrobial resistance varied within related MRSA populations and across countries. In time-scaled haplotypic density analysis, fluoroquinolone, macrolide and mupirocin resistance were associated with MRSA success, whereas gentamicin, rifampicin and trimethoprim resistance were associated with sporadicity. Usage of antimicrobials across 29 European countries varied substantially, and β-lactam, fluoroquinolone, macrolide and aminoglycoside use correlated with MRSA incidence. DiscussionOur results are the strongest yet to associate MRSA antibiotic resistance profiles and antibiotic usage with the incidence of infection and successful clonal spread, which varied by country. Harmonized isolate collection, typing, resistance profiling and alignment with antimicrobial usage over time will aid comparisons and further support country-specific interventions to reduce MRSA burden.