Bacterial Forms Research Articles

The obligate intracellular bacterium Orientia tsutsugamushi differentiates into a developmentally distinct extracellular state

Orientia tsutsugamushi (Ot) is an obligate intracellular bacterium in the family Rickettsiaceae that causes scrub typhus, a severe mite-borne human disease. Its mechanism of cell exit is unusual amongst Rickettsiaceae, as Ot buds off the surface of infected cells enveloped in plasma membrane. Here, we show that Ot bacteria that have budded out of host cells are in a distinct developmental stage compared with intracellular bacteria. We refer to these two stages as intracellular and extracellular bacteria (IB and EB, respectively). These two forms differ in physical properties: IB is both round and elongated, and EB is round. Additionally, IB has higher levels of peptidoglycan and is physically robust compared with EB. The two bacterial forms differentially express proteins involved in bacterial physiology and host-pathogen interactions, specifically those involved in bacterial dormancy and stress response, and outer membrane autotransporter proteins ScaA and ScaC. Whilst both populations are infectious, entry of IB Ot is sensitive to inhibitors of both clathrin-mediated endocytosis and macropinocytosis, whereas entry of EB Ot is only sensitive to a macropinocytosis inhibitor. Our identification and detailed characterization of two developmental forms of Ot significantly advances our understanding of the intracellular lifecycle of an important human pathogen.

Surface Management in Prevention of Nosocomial Infections in Radiography Practice

Background: For prevention of nosocomial infections in Radiography practice, it is imperative to disinfect surfaces used by all patients with appropriate disinfectants.Objective: To determine the disinfectants that are most active in eliminating micro-organisms identified on hard surfaces of equipment and accessories in the radiology department of a tertiary hospital. Methodology: Efficacy of Savlon (chlorohexidine), Jik (Sodium hydrochlorite), and Dettol (Chloroxylenol) were tested. Each examined surface was divided into three horizontal parts. Three BS medichem swabs were moistened with distilled water and used to collect sample along each surface at different points. The surfaces were cleaned with each undiluted disinfectant and allowed to dry for 20 minutes. Three different moistened swabs were used to collect another fresh sample. Chocolate agar was used to prepare the samples and later incubated in incumaxTM incubator for 24 hours at 370C. Colonies were identified under national light microscope.Results: Colony count before disinfection was one 150. After cleaning, it reduced to 60. The isolated organisms were: Staphycoccus aureus, Candida albicans, Pseudomonas aeruginosa coliform and Escherichia coli. Staphylococcus aureus which had the highest colony count of 51 before disinfection was reduced to 14 after disinfection. Escherichia coli had the least colony count before and after disinfection. Jik (sodium hydrochlorite) reduced the total bacterial load form 51 to 14 counts.Conclusion: Sodium hypochlorite (Jik) is the most effective disinfectant noticed in our centre.

Plant growth promoting rhizobacterial diversity in potato grown soil in the Gwalior region of India

There seems to be meager studies with regards to rhizo and non-rhizo microbial association with potato plant from the central India. Present study was undertaken to evaluate the microbial diversity of rhizospheric and non-rhizospheric isolates from three varieties of potato viz Kufri sindhuri, Kufri lauvkar and Kufri chipsona-3 procured from the Central Potato Research Station, Maharajpura, Gwalior. A total of 130 bacterial forms were isolated, and amongst these forty isolates were further characterized on their morphological basis, and those showing some of PGPR characteristics were identified to species level using VITEK-2 method. Various bacterial populations were found in potato rhizosphere and dominant presence was those of Bacillus subtilis, Bacillus Megaterium and Lysinibacillus sphaericus. The non-rhizospheric soil was dominant in the forms like Aeromonas salmonicida, Morxella group and Bacillus coagulans. Highest bacterial diversity was found in the rhizosphere soil of different potato cultivars than in the non-rhizospheric soil of potato.

The universally conserved nucleotides of the small subunit ribosomal RNAs.

The ribosomal RNAs, along with their substrates the transfer RNAs, contain the most highly conserved nucleotides in all of biology. We have assembled a database containing structure-based alignments of sequences of the small-subunit rRNAs from organisms that span the entire phylogenetic spectrum, to identify the nucleotides that are universally conserved. In its simplest (bacterial and archaeal) forms, the small-subunit rRNA has ∼1500 nt, of which we identify 140 that are absolutely invariant among the 1961 species in our alignment. We examine the positions and detailed structural and functional interactions of these universal nucleotides in the context of a half century of biochemical and genetic studies and high-resolution structures of ribosome functional complexes. The vast majority of these nucleotides are exposed on the subunit interface surface of the small subunit, where the functional processes of the ribosome take place. However, only 40 of them have been directly implicated in specific ribosomal functions, such as contacting the tRNAs, mRNA, or translation factors. The roles of many other invariant nucleotides may serve to constrain the positions and orientations of those nucleotides that are directly involved in function. Yet others can be rationalized by participation in unusual noncanonical tertiary structures that may uniquely allow correct folding of the rRNA to form a functional ribosome. However, there remain at least 50 nt whose universal conservation is not obvious, serving as a metric for the incompleteness of our understanding of ribosome structure and function.

Comparing the Effect of Tea Tree Oil and Lavender on Bacterial Samples of Nurses’ Hands

Objective: One of the most typical ways of transmitting the infection is by hand; thus, much emphasis has been placed on hand hygiene. In this regard, this study aimed to determine the effect of tea tree oil and lavender on bacterial samples found in nurses’ hands. Methods: This quasi-experimental study was performed on all nurses working in ICU wards. The samples were selected by an easy non-probability sampling method. The research tools were demographic information questionnaires and a bacterial culture registration form. In the intervention phase, on the first day in the first experimental group, 2 drops of lavender oil, on the second day, 2 drops of tea tree oil, and on the third day, 2 drops of distilled water were poured on the nurse’s palm, and their hands were massaged for 10 seconds. Before and after the intervention, culture samples were taken from each group. The obtained data were analyzed with the Chi-square test, paired t test, and ANOVA in SPSS software v. 20 at a significant level of less than 0.05. Results: The most common organisms isolated at the beginning of the study were Staphylococcus epidermidis (76.8%), Staphylococcus aureus (8.8%), Klebsiella spp. (7.9%), and Escherichia coli (6.5%). The use of lavender oil and tea tree has been effective in reducing the microbial load of the palm. Hand massage with lavender oil had less effect on reducing germs (P=0.003), but tea tree oil had a more significant reduction of pathogens (P=0.001). Conclusion: Lavender and tea tree oil reduced the microbial load of nurses. Both methods have been effective, but the effect of tea tree oil was more significant. Therefore, their use in handwashing is recommended to eliminate palm bacteria in ICU staff.

Determination of Bacterial Concrete Strength Using Bacillus Subtilis and Lightweight Expandable Clay Aggregate

This study examines the impact of bacterial concrete on strength and self-healing. Bacterial concrete has better compressive strength, permeability, corrosion resistance, chemical precursors, alkalinity resistance, and mechanical stress. Bacillus subtilis calcium lactate and spore powder effects are explored in this study, and the influence of this bacterial form on strength and self-healing capacity to crack repair. The Bacillus subtilis concentration 105 cells/mL is used in concrete with calcium lactate 0.3% of cement. In another trial, calcium lactates 0.3% and spore powder 0.5% of cement with Bacillus subtilis concentration of 105 cells/mL and lightweight expandable clay aggregate (LECA) is 30% replaced to the coarse aggregate used in concrete respectively. The conventional concrete and bacterial concrete cubes were molded with dimensions of 150 mm x 150 mm x 150 mm, cylinders with dimensions of 100 mm x 200 mm, and a beam with dimensions of 100 mm x 100 mm x 500 mm. These specimens were evaluated after 7 and 28 days of cure. The compressive, split tensile, and flexural strength of bacterial concrete was raised by 23%, 8%, and 7%, respectively when compared to conventional concrete. Thus, the experimental findings reveal that Bacillus subtilis at 105 cells/ml cells with 0.3% calcium lactate has a substantial impact on the strength and self-healing of bacterial concrete.

Aspiration pneumonia complicated by disseminated fungal sepsis in the setting of incidental esophagitis cystica: A rare autopsy case report with review of the literature

Abstract Introduction/Objective Esophageal retention cysts are presumed to arise from obstruction of the excretory ducts of the submucosal glands of the esophagus. Since the first description by Kuhne in 1899, different terminologies have been used to denote these lesions including retention cysts, mucocele, esophagitis cystica, and cyst of esophageal submucosal gland duct. Esophageal retention cysts are generally benign, asymptomatic and discovered incidentally. However, a few studies have reported symptomatic dysphagia and to date, only one autopsy case report described esophageal retention cysts contributing to aspiration pneumonia and cause of death. Methods/Case Report We present a case of aspiration pneumonia, associated with both reduced oropharyngeal tone secondary to alcohol consumption and esophageal dysphagia due to esophagitis cystica in a 69 year old female with alcohol use disorder complicated by alcoholic liver disease. The patient was found unresponsive at home with bloody oral secretions and evidence of head trauma. On autopsy all lung lobes demonstrated that large and small airways and alveoli were filled with vegetable matter, associated with prominent bacterial and fungal forms. EVG stain demonstrated vascular disruption with red blood cell extravasation, next to a focus of aspiration pneumonia. This explained the patient’s “bloody oral secretions”, originating from the respiratory tract. Demonstration of fungal hyphae in sections from lungs as well as necrotic colon was consistent with disseminated fungal sepsis. While sections from esophagus showed no varices which could explain the patient’s “bloody oral secretions” in the setting of alcoholic liver disease, there were numerous dilated esophageal submucosal cysts. Cysts were lined by a single to double layer of cuboidal epithelium with eosinophilic cytoplasm and basally-located, bland-appearing nuclei. Conclusion In summary, we present a rare case of esophageal retention cyst associated aspiration pneumonia complicated by pulmonary hemorrhage, adding to the growing body of knowledge regarding fatal complications of these usually incidental lesions.

Cadaverine biosynthesis contributes to decreased Escherichia coli susceptibility to antibiotics

Some bacterial stress responses are involved in survival under antibiotic treatment and contribute to less susceptible microbial forms selection. Here, we tested the role of cadaverine, one of the biogenic polyamines considered as universal adaptogens, in the processes. The expression of ldcC and cadA genes, encoding cadaverine-producing lysine decarboxylase, increased in Escherichia coli cells exposed to β-lactams and fluoroquinolones but not aminoglycosides. The transcriptional regulators RpoS and SoxS controlled the expression of ldcC and cadA, respectively, in response to antibiotics. Exogenous cadaverine had little effect on E. coli antibiotic susceptibility, whereas non-antibiotic-induced endogenous cadaverine contributed to its tolerance to β-lactams, fluoroquinolones, and aminoglycosides. Antibiotic-induced cadaverine synthesis promoted bacterial survival under fluoroquinolone exposure, as well as could contribute to low-resistant bacterial forms development. Selection under the fluoroquinolone levofloxacin exposure toward bacteria with an increased ability to synthesize cadaverine and negative correlation between LdcC activity and fluoroquinolone susceptibility in the selected forms were demonstrated. The same correlation in a special group of low-level resistant clinical E. coli isolates was revealed. So, cadaverine biosynthesis appeared to be a significant player in decreased E. coli antibiotic susceptibility development.

Seven-day Oral Intake of Orthosiphon stamineus Leaves Infusion Exerts Antiadhesive Ex Vivo Activity Against Uropathogenic E. coli in Urine Samples.

Orthosiphon stamineus leaves (Java tea) extract is traditionally used for the treatment of urinary tract infections. According to recent in vitro data, animal infection studies, and transcriptomic investigations, polymethoxylated flavones from Java tea exert antiadhesive activity against uropathogenic Escherichia coli (UPEC). This antiadhesive activity has been shown to reduce bladder and kidney lesion in a mice infection model. As no data on the antivirulent activity of Java tea intake on humans are available, a biomedical study was performed on 20 healthy volunteers who self-administered Orthosiphon infusion (4 × 3 g per day, orally) for 7 days. The herbal material used for the study conformed to the specification of the European Pharmacopoeia, and ultra high-performance liquid chromatography (UHPLC) of the infusion showed rosmarinic acid, caffeic acid, and cichoric acid to be the main compounds aside from polymethoxylated flavones. Rosmarinic acid was quantified in the tea preparations with 243 ± 22 µg/mL, indicating sufficient reproducibility of the preparation of the infusion. Urine samples were obtained during the biomedical study on day 1 (control urine, prior to Java tea intake), 3, 6 and 8. Antiadhesive activity of the urine samples was quantified by flowcytometric assay using pre-treated UPEC NU14 and human T24 bladder cells. Pooled urine samples indicated significant inhibition of bacterial adhesion on day 3, 6 and 8. The urine samples had no influence on the invasion of UPEC into host cells. Bacterial proliferation was slightly reduced after 24 h incubation with the urine samples. Gene expression analysis (qPCR) revealed strong induction of fitness and motility gene fliC and downregulation of hemin uptake system chuT. These data correlate with previously reported datasets from in vitro transcriptomic analysis. Increased bacterial motility was monitored using a motility assay in soft agar with UPEC UTI89. The intake of Java tea had no effect on the concentration of Tamm-Horsfall Protein in the urine samples. The present study explains the antiadhesive and anti-infective effect of the plant extract by triggering UPEC from a sessile lifestyle into a motile bacterial form, with reduced adhesive capacity.

Biomineralization processes in modern calcareous tufa: Possible roles of viruses, vesicles and extracellular polymeric substances (Corvino Valley – Southern Italy)

AbstractA modern lithifying biofilm, associated with fluvial calcareous tufa in southern Italy, is composed of a complex community of micro‐organisms including autotrophic and heterotrophic bacteria, together with likely viruses (bacteriophages) and bacterial vesicles. Produced by most of the bacterial forms, extracellular polymeric substances are widespread and are the main sites of mineral precipitation, rather than the microbial cells themselves, but viruses and vesicles also appear to be mineralized. Neoformed biominerals replace these organic substrates starting with an amorphous phase rich in Ca, Si and several other cations followed by two different types of proto‐crystal: a granular−irregular Ca‐rich type and a lamellar/fibrous Si‐Al‐Mg‐rich type. Their growth leads to the formation of Ca‐carbonate and Mg‐silicate crystals that preserve their initial different habits. The coeval precipitation of silicate and carbonate at the initial stage of bio‐mediated mineral formation is a process common within freshwater and saline to hypersaline‐water calcareous microbialites. This is most probably related to the initial formation of poorly‐crystallized Mg‐Si phases that precipitate within the extracellular polymeric substances, related to the rise of pH in the zone of active oxygenic photosynthesis, and the mineralization of viruses, associated with the successive nucleation of Ca‐carbonate in close proximity to bacterial degradation of organic matter.

Comparative efficiency of various molecular genetic methods in diagnosing tuberculosis

Objective – to compare the efficacy of various molecular genetic methods for diagnosing tuberculosis and determining drug susceptibility to rifampicin (RIF). Materials and Methods. We conducted a retrospective study of the sputum analysis results on 1,992 patients with pulmonary tuberculosis treated at Saratov Oblast Clinical Tuberculosis (TB) Dispensary from 2014 through 2018. The following methods were used: real-time polymerase chain reaction (PCR), biological microarrays, automated Xpert® MTB/RIF technology. Statistical processing of the research results was carried out using the Bayes formula based on contingency tables (four-field table) and the χ2 test. When evaluating the significance of differences between relative values, we employed the critical significance level of 0.05. Results. In terms of etiological diagnosis of TB, higher diagnostic sensitivities of the real-time PCR and biological microarray methods (73.9% and 70.3%, correspondingly) were established, as compared with the Xpert® MTB/RIF method (34.2%) (p<0.001). The sensitivity of all methods depended on the massiveness of bacterial excretion and clinical form of TB. Conclusions: The Xpert MTB/RIF method exhibited lower diagnostic sensitivity in verifying the diagnosis of TB, whereas its operational characteristics in terms of determining RIF-resistance were sufficiently high (sensitivity at 89.7%, specificity at 89.1%, and efficacy at 89.4%), which was comparable with the characteristics of biological microarray method (93.9%, 71.8%, 82.9%; p=0.127, p<0.001, p=0.139, respectively).

Post-treatment bacterial endocarditis mimicking fungal organisms: a morphologic comparison and tips for avoiding this diagnostic pitfall

BackgroundHistopathologic differentiation of bacterial endocarditis from yeast-like fungal endocarditis is usually straightforward; however, an underappreciated phenomenon is the effect of antimicrobial therapy on bacterial size, shape and septa (cross-wall) formation resulting in bacterial forms that mimic yeast-like fungi. In this article we illustrate the alterations that occur in antibiotic-treated Staphylococcus aureus endocarditis and compare these changes to histopathologic findings in unaltered S. aureus and Histoplasma endocarditis, respectively. MethodsResected valves from three cases of endocarditis were compared based on the type ofinflammatory reaction, organism morphology and culture results. Case 1 was S. aureus endocarditis initially misclassified as Histoplasma due to its atypical morphologic and histopathologic features. The two cases included for comparison were an S. aureus endocarditis with more classic features and an Histoplasma capsulatum endocarditis. Hematoxylin and eosin (H&E), Gram, periodic acid Schiff (PAS), Gomori-Grocott methenamine silver stains (GMS), and culture results were compared in all cases. Molecular and immunohistochemistry tests were used for confirmation of first case. High power oil-immersion was used to visualize organisms’ characteristics in all three cases. ResultsCase 1 and Case 3 (Histoplasma-infected valves) had fibrinous exudates with scattered macrophages. The microorganisms observed in the first case of methicillin-sensitive S. aureus (MSSA) were ∼ 2-3 μm by GMS stain and had prominent septations. Histoplasma yeast were round to oval, ∼ 3-4 μm in size and demonstrated budding. S. aureus without alterations were round, ∼ 1 μm in size, and lacked prominent septations. Necrotizing purulent inflammation was present in the unaltered case of MSSA. The MSSA case with alterations from antibiotic treatment did not stain well with the Gram stain and organisms were best visualized with the PAS and GMS stains. ConclusionsAntibiotic therapy for bacterial endocarditis can alter the inflammatory reaction to infection, bacterial size, septa formation, and staining characteristics. Knowledge of these therapy-related effects and use of high-power magnification helps to avoid misclassification as yeast-like fungi.

Agro-ecological Characterization of Vermicomposted Sewage Sludge from Municipal and Poultry Enterprise Wastewater Treatment Plants

The purpose of this study was to make an agro-ecological characterization of vermicompost (VC) produced from sewage sludge (SS). As a substrate, SS from municipal and poultry meat processing enterprise wastewater treatment plants (MTP and PTP, respectively) was utilized. The substrates were vermicomposted by Red Californian earthworm (Lumbricus rubellus) for 120 days. For VC quality assessment, 19 physicochemical and 6 microbiological parameters were used. The evaluation of physicochemical parameters was done according to ISO standard methods and microbiological analysis-by plating 1 mL of sample dilutions on selective, chromogenic culture medium sheets. It was found that the vermicompost from MTP (VC-M) had higher levels of EC, mineral elements (N, P and K compounds in forms available to plants), heavy metals (Mn, Cu, Zn, Cr, Ni, Pb, Cd) and coliforms, and lower levels of pH, TOC, C/N ratio, Fe, total plate count (TPC), Enterobacteriaceae and Salmonella spp. counts compared to VC from PTP (VC-P). During the vermicomposting process, the substrates from both wastewater treatment plants (TPs) showed similar trends towards decrease in pH, TOC, N-NH4+, C/N ratio, TPC, coliforms, E. coli, Enterobacteriaceae and Salmonella spp. counts, whereas the opposite trends were established for EC, TKN, N-NO3-, TP, P2O5, TK, and K2O values. The vermicomposting had a negligible effect on heavy metal concentrations. In the final substrates E. coli were not detected, while the bacterial spore forms (Clostridium perfringens) were not eliminated. The final substrates cannot be used as fertilizers or soil amendments because of the presence of Salmonella spp. and C. perfringens over the permissible limits according to EU and Bulgarian regulations.

Differential Early in vivo Dynamics and Functionality of Recruited Polymorphonuclear Neutrophils After Infection by Planktonic or Biofilm Staphylococcus aureus.

Staphylococcus aureus is a human pathogen known for its capacity to shift between the planktonic and biofilm lifestyles. In vivo, the antimicrobial immune response is characterized by the recruitment of inflammatory phagocytes, namely polymorphonuclear neutrophils (PMNs) and monocytes/macrophages. Immune responses to planktonic bacteria have been extensively studied, but many questions remain about how biofilms can modulate inflammatory responses and cause recurrent infections in live vertebrates. Thus, the use of biologically sound experimental models is essential to study the specific immune signatures elicited by biofilms. Here, a mouse ear pinna model of infection was used to compare early innate immune responses toward S. aureus planktonic or biofilm bacteria. Flow cytometry and cytokine assays were carried out to study the inflammatory responses in infected tissues. These data were complemented with intravital confocal imaging analyses, allowing the real-time observation of the dynamic interactions between EGFP + phagocytes and bacteria in the ear pinna tissue of LysM-EGFP transgenic mice. Both bacterial forms induced an early and considerable recruitment of phagocytes in the ear tissue, associated with a predominantly pro-inflammatory cytokine profile. The inflammatory response was mostly composed of PMNs in the skin and the auricular lymph node. However, the kinetics of PMN recruitment were different between the 2 forms in the first 2 days post-infection (pi). Two hours pi, biofilm inocula recruited more PMNs than planktonic bacteria, but with decreased motility parameters and capacity to emit pseudopods. Inversely, biofilm inocula recruited less PMNs 2 days pi, but with an “over-activated” status, illustrated by an increased phagocytic activity, CD11b level of expression and ROS production. Thus, the mouse ear pinna model allowed us to reveal specific differences in the dynamics of recruitment and functional properties of phagocytes against biofilms. These differences would influence the specific adaptive immune responses to biofilms elicited in the lymphoid tissues.

Ability of Procalcitonin and C-Reactive Protein for Discriminating between Bacterial and Enteroviral Meningitis in Children Using Decision Tree

Bacterial meningitis (BM) is a public health burden in developing countries, including Central Asia. This disease is characterized by a high mortality rate and serious neurological complications. Delay with the start of adequate therapy is associated with an increase in mortality for patients with acute bacterial meningitis. Cerebrospinal fluid culture, as a gold standard in bacterial meningitis diagnosis, is time-consuming with modest sensitivity, and this is unsuitable for timely decision-making. It has been shown that bacterial meningitis differentiation from viral meningitis could be done through different parameters such as clinical signs and symptoms, laboratory values, such as PCR, including blood and cerebrospinal fluid (CSF) analysis. In this study, we proposed the method for distinguishing the bacterial form of meningitis from enteroviral one. The method is based on the machine learning process deriving making decision rules. The proposed fast-and-frugal trees (FFTree) decision tree approach showed an ability to determine procalcitonin and C-reactive protein (CRP) with cut-off values for distinguishing between bacterial and enteroviral meningitis (EVM) in children. Such a method demonstrated 100% sensitivity, 96% specificity, and 98% accuracy in the differentiation of all cases of bacterial meningitis in this study. These findings and proposed method may be useful for clinicians to facilitate the decision-making process and optimize the diagnostics of meningitis.

Investigation of biological activity of recultivated iron ore processing wastes of Abagur agglomeration and processing plant, Novokuznetsk

The problem of biotransformation of man-caused disturbed lands is of great importance for the restoration of soil fertility. The leading role in this process is played by soil microorganisms. Toxic industrial waste can make negative adjustments to the species composition of soil microflora (can cause mutation of many microorganisms, as well as the death of some of them). Under certain conditions, in the presence of harmful substances, new microorganisms can arise that decompose precisely those substances that cause soil pollution. Consequently, a new microflora appears, capable of carrying out its trophic functions on an inductive basis on industrially contaminated substrates. As a man-caused control, a mineral substrate of the tailing dump of Abagurskaya agglomeration and processing plant JSC, unfavorable for the vital activity of soil microflora, where, due to the absence of plant residues and other carbon-containing components, an extremely low level of intensity of enzymatic hydrolytic processes was used. The introduction of organic matter containing sewage sludge (WWS) into the substrate as a recultivator creates favorable conditions for the formation of a stable vegetation cover. The presence of an organic component in the mineral substrate of the iron-containing tailings waste creates favorable conditions for the vital activity of microflora. Carbon-containing compounds of technozems create a certain pool of microorganisms. The products of their vital activity create the basis for the formation of an organic-mineral substrate, favorable conditions for the growth of higher plants and the creation of a stable phytocenosis. For the induced manifestation of enzymatic activity under these extreme conditions, a considerable time is required for the bacterial forms introduced from the outside to be able to fix their presence. The maximum activity, comparable both after 1 month and after 3 months with the natural control, was noted on the experimental site with the placement of WWS as a soil conditioner. The presence of WWS and intensive accumulation of leaf litter and other plant residues contribute to the favorable development of cellulose-degrading bacteria, among which rod-shaped forms of the genus Clostridium predominate. The quantitative indicators of the enzymatic activity of technozems are close to the natural level and significantly exceed those of technogenic control.

EVIDENCE BASED MANAGEMENT OF UPAPLUTA YONIVYAPAD – IN VITRO CASE STUDY REPORT

Vaginitis is a common problem during pregnancy. Most commonly Candidial vaginitis & Bacterial vaginitis forms are found. It has hazardous effects on pregnancy like miscarriage, preterm labour etc. Therefore, careful manage- ment of patient is necessary. Ayurveda described this condition under Yonivyapad chapter which labelled as Upapluta Yonivyapad which means overflowed vaginal discharge with vaginal itching & mild abdominal pain. We can correlate Upapluta Yonivyapad with Candidial vaginitis. In this case study we have taken 2 cases of Candidial Vaginitis & local application of herbal drugs kwath was done. Evidence based beneficial results shown through HPE to the patient of Upapluta Yonivyapad & is safe during pregnancy. By this we can conclude that the ayurvedic drugs is the best solution to treat vaginitis during pregnancy. Keywords: Vaginitis, Upapluta Yonivyapad.

The Natural Alkaloid Berberine Can Reduce the Number of Pseudomonas aeruginosa Tolerant Cells.

The eradication of recurrent Pseudomonas aeruginosa (PA) lung infection in cystic fibrosis (CF) patients may be hampered by the development of persistent bacterial forms, which can tolerate antibiotics through efflux pump overexpression. After demonstrating the efflux pump inhibitory effect of the alkaloid berberine on the PA MexXY-OprM efflux pump, in this study, we tested its ability (80/320 μg/mL) to enhance tobramycin (20xMIC/1000xMIC) activity against PA planktonic/biofilm cultures. Preliminary investigations of the involvement of MexY in PA tolerance to tobramycin treatment, performed on the isogenic pair PA K767 (wild type)/K1525 (ΔmexY) growing in planktonic and biofilm cultures, demonstrated that the ΔmexY mutant K1525 produced a lower (100 and 10 000 times, respectively) amount of tolerant cells than that of the wild type. Next, we grew broth cultures of PAO1, PA14, and 20 PA clinical isolates (of which 13 were from CF patients) in the presence of 20xMIC tobramycin with and without berberine 80 μg/mL. Accordingly, most strains showed a greater (from 10- to 1000-fold) tolerance reduction in the presence of berberine. These findings highlight the involvement of the MexXY-OprM system in the tobramycin tolerance of PA and suggest that berberine may be used in new valuable therapeutic combinations to counteract persister survival.

Genetic individuality and interspecies altruism: modelling symbiogenesis using different types of symbiotic bacteria

In this minireview, we address the trade-off between biological altruism (group adaptation result-ing from the ability of an organism to improve the fitness of an associate at the expense of its own fitness) and symbiogenesis — the evolutionary pathway based on genetic integration of non-related species. We address symbiogenesis as a multi-stage process, which involves for-mation of superspecific hereditary systems — functionally integral symbiogenomes (under the facultative partners’ interactions) reorganized into the structurally integral hologenomes (in the obligatory symbioses). The best studied case of symbiogenesis is represented by the evolution of the eukaryotic cell based on transformation of symbiotic bacteria into cellular organelles. This evolution is associated with the deep reduction of microsymbionts’ genomes and with allocation of their genes into the hosts. As a result, microsymbionts lost their Genetic INdividuality (GIN), characterized by an ability to implement DNA- and RNA-based template syntheses required for genome maintenance and expression. Under facultative symbiotic dependence on hosts, the par-tial loss of GIN is due to a “symbiont → host” altruism which in the N2-fixing microbe–plant symbioses results in formation of non-reproducible bacterial forms (e.g., intracellular bacteroids in rhizobia or multiple heterocysts in Nostoc). If micro-symbionts lose their ability of autonomous existence (e.g., in the vertically transmitted intracellular symbionts), they are switched to the “forced altruism” in which the GIN reduction is required for the stable persistence of symbionts in hosts. Therefore, organellogenesis involves the sequential increase of the symbionts’ de-pendency on hosts: conditional → facultative → obligatory → absolute. It is associated with the reorganization of microbes into semi-autonomous cellular components, which may be completely devoid of their own genomes.

Split conformation of Chaetomium thermophilum Hsp104 disaggregase

Hsp104 and its bacterial homolog ClpB form hexameric ring structures and mediate protein disaggregation. The disaggregated polypeptide is thought to thread through the central channel of the ring. However, the dynamic behavior of Hsp104 during disaggregation remains unclear. Here, we reported the stochastic conformational dynamics and a split conformation of Hsp104 disaggregase from Chaetomium thermophilum (CtHsp104) in the presence of ADP by X-ray crystallography, cryo-electron microscopy (EM), and high-speed atomic force microscopy (AFM). ADP-bound CtHsp104 assembles into a 65 left-handed spiral filament in the crystal structure at a resolution of 2.7Å. The unit of the filament is a hexamer of the split spiral structure. In the cryo-EM images, staggered and split hexameric rings were observed. Further, high-speed AFM observations showed that a substrate addition enhanced the conformational change and increased the split structure's frequency. Our data suggest that split conformation is an off-pathway state of CtHsp104 during disaggregation.