HISTOLOGICAL STUDY OF LETTUCE AND BASIL BY INFECTION WITH PS. AERUGINOSA AND PS. FLUORESCENS IN VITRO

2020 ◽  
pp. 56-69
Author(s):  
G.V. GODOVA ◽  
◽  
A.A. OVOD ◽  
N.V. ASTAKHOVA ◽  
E.A. KALASHNIKOVA ◽  
...  
Keyword(s):  
Virulent Strain ◽  
Histological Study ◽  
Plant Cells ◽  
Ultrastructural Changes ◽  
Cytotoxic Action ◽  
Biochemical Tests ◽  
Potential Threat ◽  
Gram Staining ◽  
Electronic Microscope

Studies based on the use of an electronic microscope revealed ultrastructural changes in plant cells of lettuce and Basil in interaction with Ps.aeruginosa ATCC B3994, expressed in deformation and violation of the integrity of the cell wall, increasing the size of chloroplasts, sometimes full destruction accompanied by the increased number of plastoglobuli. Ps.fluorescens ATСC948 were localized in the cytoplasm and chloroplasts of cells without a cytotoxic action. Plant isolates were identified with gram staining, biochemical tests and PCR. Contamination of non-heattreated leaf vegetables with virulent strain Ps. aeruginosa is a potential threat to human health and life.

scholarly journals Bacteriocin-mediated inhibition of some common pathogens by wild and mutant Lactobacillus species and in vitro amplification of bacteriocin encoding genes

ADMET & DMPK ◽  
2021 ◽  
Author(s):  
Arooj Ahsan ◽  
Bushra Mazhar ◽  
Muhammad Kamran Khan ◽  
Madiha Mustafa ◽  
Muhammad Hammad ◽  
...  
Keyword(s):  
Gene Sequence ◽  
Diffusion Method ◽  
Lactobacillus Species ◽  
Lactobacillus Curvatus ◽  
Biochemical Tests ◽  
Specific Primers ◽  
Gram Staining ◽  
Salmonella Enteritis ◽  
Encoding Genes

Lactobacilli are the most common probiotics used in food and other industries because of their capability of producing bacteriocins. Bacteriocins are compounds that are used to kill pathogenic microorganisms. As most bacteria have become resistant to synthetic antibacterial tools, the importance of using probiotics as antibacterial agents has increased. This work was done to check the bacteriocin effect on some common pathogens and the influence of mutation on the bacteriocin activity of Lactobacilli was also investigated. Four strains were isolated, identified from meat and pickles samples via culturing methods, staining, biochemical tests, and ribotyping. Preliminary tests, including Gram staining and catalase test, were done for the confirmation of Lactobacillus species. All strains were gram-positive and catalase-negative. Antibacterial activity was checked against Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus thuringiensis, Escherichia coli, and Salmonella enteritis via agar well diffusion method. The mutations were done using ethidium bromide and the influence of wild and mutants were also checked. Interestingly, mutants developed more virulence than wild ones. It was also observed that they all were sensitive to pepsin. Protein estimation was done via Bradford method. Ribotyping of GCU-W-PS1 revealed 99 % homology with Lactobacillus plantarum and GCU-W-MS1 to Lactobacillus curvatus (99 % homology). Curvacin A, sakacin P, and plantaricin A genes were also amplified using specific primers. Gene sequence showed the presence of curvacin A gene in GCU-W-MS1. It was concluded that lactic acid bacteria could be used as antibacterial tools against common pathogens.

scholarly journals Isolation of Lactobacillus species from fermented Parkia biglobosa seed and screening for their probiotic activity

2021 ◽  
Vol 13 (Volume 13, Issue 2) ◽  
pp. 212-216
Author(s):  
K. Kuti ◽  
I.M. Hussaini ◽  
A. Usman ◽  
A. Isa
Keyword(s):  
Antibacterial Activity ◽  
Low Ph ◽  
Lactobacillus Species ◽  
Biochemical Tests ◽  
Antimicrobial Substances ◽  
Probiotic Potential ◽  
Gram Staining ◽  
Colonial Morphology ◽  
Lactobacillus Spp

Abstract. Food fermentation has been a tradition for decades due to its immense benefits. Lactic acid bacteria are known to possess probiotic potential due to various chemical antimicrobial substances they produce. This research was aimed at isolating Lactobacillus spp. from locally fermented locust beans and screening the isolates for their probiotic potential. A total of 20 samples of fermented locust beans (P. biglobosa) were collected and processed. The samples were inoculated onto de-Mann Rogosa Sharpe (MRS) media and incubated anaerobically. The isolates with characteristic colonial morphology of Lactobacillus sp. were characterized using Gram staining and biochemical tests. Lactobacillus spp. isolates were screened in-vitro for probiotic potential using the following parameters, tolerance of low pH, bile tolerance and antimicrobial activity. Four isolates of Lactobacillus species consisting of Lactobacillus fermentum and Lactobacillus plantarum were isolated from different fermented locust beans. All isolates showed tolerance to low pH and bile concentration of 3.04 and 0.3%, respectively. The Lactobacillus species isolates showed varying antibacterial activity against all test bacteria with isolate S06 (L. plantarum) showing the highest degree of antibacterial activity. In conclusion, the Lactobacillus species isolated from fermented locust beans can serve as probiotic candidate.

Ultrastructural Changes in Tumor Cells During Human Peripheral Blood Monocyte-Mediated Cytotoxicity

1981 ◽  
Vol 39 ◽  
pp. 452-453
Author(s):  
K. E. Muse ◽  
D. G. Fischer ◽  
H. S. Koren
Keyword(s):  
Target Cell ◽  
Human Peripheral Blood ◽  
Mononuclear Phagocytes ◽  
Ultrastructural Changes ◽  
Target Cells ◽  
Limited Information ◽  
Blood Monocytes ◽  
Tumoricidal Activity ◽  
Activated State

Mononuclear phagocytes, a pluripotential cell line, manifest an array of basic extracellular functions. Among these physiological regulatory functions is the expression of spontaneous cytolytic potential against tumor cell targets.The limited observations on human cells, almost exclusively blood monocytes, initially reported limited or a lack of tumoricidal activity in the absence of antibody. More recently, freshly obtained monocytes have been reported to spontaneously impair the biability of tumor target cells in vitro (Harowitz et al., 1979; Montavani et al., 1979; Hammerstrom, 1979). Although the mechanism by which effector cells express cytotoxicity is poorly understood, discrete steps can be distinguished in the process of cell mediated cytotoxicity: recognition and binding of effector to target cells,a lethal-hit stage, and subsequent lysis of the target cell. Other important parameters in monocyte-mediated cytotoxicity include, activated state of the monocyte, effector cell concentrations, and target cell suseptibility. However, limited information is available with regard to the ultrastructural changes accompanying monocyte-mediated cytotoxicity.

Ultrastructural Changes in Three Different Mammalian Cells Following Ultraviolet Laser Irradiation

1972 ◽  
Vol 30 ◽  
pp. 350-351
Author(s):  
K. Shankar Narayan ◽  
Kailash C. Gupta ◽  
Tohru Okigaki
Keyword(s):  
Ultraviolet Light ◽  
Mammalian Cells ◽  
Biological Effects ◽  
Survival Rates ◽  
Chinese Hamster ◽  
Cell Types ◽  
Differential Sensitivity ◽  
Ultrastructural Changes ◽  
Ultraviolet Laser

The biological effects of short-wave ultraviolet light has generally been described in terms of changes in cell growth or survival rates and production of chromosomal aberrations. Ultrastructural changes following exposure of cells to ultraviolet light, particularly at 265 nm, have not been reported.We have developed a means of irradiating populations of cells grown in vitro to a monochromatic ultraviolet laser beam at a wavelength of 265 nm based on the method of Johnson. The cell types studies were: i) WI-38, a human diploid fibroblast; ii) CMP, a human adenocarcinoma cell line; and iii) Don C-II, a Chinese hamster fibroblast cell strain. The cells were exposed either in situ or in suspension to the ultraviolet laser (UVL) beam. Irradiated cell populations were studied either "immediately" or following growth for 1-8 days after irradiation.Differential sensitivity, as measured by survival rates were observed in the three cell types studied. Pattern of ultrastructural changes were also different in the three cell types.

In Vitro, In Silico and Ex Vivo Studies of Dihydroartemisinin Derivatives as Antitubercular Agents

2019 ◽  
Vol 19 (8) ◽  
pp. 633-644 ◽  
Author(s):  
Komal Kalani ◽  
Sarfaraz Alam ◽  
Vinita Chaturvedi ◽  
Shyam Singh ◽  
Feroz Khan ◽  
...  
Keyword(s):  
Bone Marrow ◽  
In Silico ◽  
Ex Vivo ◽  
Virulent Strain ◽  
Infection Model ◽  
Mouse Bone Marrow ◽  
Significant Activity ◽  
Macrophage Infection ◽  
In Silico Studies

Introduction: As a part of our drug discovery program for anti-tubercular agents, dihydroartemisinin (DHA-1) was screened against Mtb H37Rv, which showed moderate anti-tubercular activity (>25.0 µg/mL). These results prompted us to carry out the chemical transformation of DHA-1 into various derivatives and study their antitubercular potential. Materials and Methods: DHA-1 was semi-synthetically converted into four new acyl derivatives (DHA-1A – DHA-1D) and in-vitro evaluated for their anti-tubercular potential against Mycobacterium tuberculosis H37Rv virulent strain. The derivatives, DHA-1C (12-O-(4-nitro) benzoyl; MIC 12.5 µg/mL) and DHA-1D (12-O-chloro acetyl; MIC 3.12µg/mL) showed significant activity against the pathogen. Results: In silico studies of the most active derivative (DHA-1D) showed interaction with ARG448 inhibiting the mycobacterium enzymes. Additionally, it showed no cytotoxicity towards the Vero C1008 cells and Mouse bone marrow derived macrophages. Conclusion: DHA-1D killed 62% intracellular M. tuberculosis in Mouse bone marrow macrophage infection model. To the best of our knowledge, this is the first-ever report on the antitubercular potential of dihydroartemisinin and its derivatives. Since dihydroartemisinin is widely used as an antimalarial drug; these results may be of great help in anti-tubercular drug development from a very common, inexpensive, and non-toxic natural product.

scholarly journals Identification of Putative Virulence Genes by DNA Methylation Studies in the Cereal Pathogen Fusarium graminearum

Cells ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 1192
Author(s):  
Francesco Tini ◽  
Giovanni Beccari ◽  
Gianpiero Marconi ◽  
Andrea Porceddu ◽  
Micheal Sulyok ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Fusarium Graminearum ◽  
Environmental Changes ◽  
Virulent Strain ◽  
Fungal Virulence ◽  
Wheat Seedlings ◽  
Wide Range ◽  
Conidia Production ◽  
Natural Hosts

DNA methylation mediates organisms’ adaptations to environmental changes in a wide range of species. We investigated if a such a strategy is also adopted by Fusarium graminearum in regulating virulence toward its natural hosts. A virulent strain of this fungus was consecutively sub-cultured for 50 times (once a week) on potato dextrose agar. To assess the effect of subculturing on virulence, wheat seedlings and heads (cv. A416) were inoculated with subcultures (SC) 1, 23, and 50. SC50 was also used to re-infect (three times) wheat heads (SC50×3) to restore virulence. In vitro conidia production, colonies growth and secondary metabolites production were also determined for SC1, SC23, SC50, and SC50×3. Seedling stem base and head assays revealed a virulence decline of all subcultures, whereas virulence was restored in SC50×3. The same trend was observed in conidia production. The DNA isolated from SC50 and SC50×3 was subject to a methylation content-sensitive enzyme and double-digest, restriction-site-associated DNA technique (ddRAD-MCSeEd). DNA methylation analysis indicated 1024 genes, whose methylation levels changed in response to the inoculation on a healthy host after subculturing. Several of these genes are already known to be involved in virulence by functional analysis. These results demonstrate that the physiological shifts following sub-culturing have an impact on genomic DNA methylation levels and suggest that the ddRAD-MCSeEd approach can be an important tool for detecting genes potentially related to fungal virulence.

scholarly journals Isolation and characterization of Pseudomonas syringae isolates affecting stone fruits and almond in Montenegro

2021 ◽  
Author(s):  
Tamara Popović ◽  
Jelena Menković ◽  
Anđelka Prokić ◽  
Nevena Zlatković ◽  
Aleksa Obradović
Keyword(s):  
Pseudomonas Syringae ◽  
Housekeeping Genes ◽  
Molecular Characteristics ◽  
Biochemical Tests ◽  
High Genetic Diversity ◽  
Leaf Petiole ◽  
Molecular Features ◽  
Isolation And Characterization ◽  
Genomic Species

AbstractIn Montenegro, stone fruit species are grown on intensive and semi-intensive commercial plantations. However, almond production is mainly organized on family gardens and for household consumption. During two seasons (2017–2018), we surveyed apricot, peach, nectarine, sweet cherry, Japanese plum, and almond orchards for the presence of bacterial diseases at different geographical locations in Montenegro. From leaf, petiole and fruit lesions, branch or twig cankers, and necrotizing buds, a total of 29 isolates were obtained and subjected to identification based on their morphological, pathogenic, biochemical, and molecular characteristics. Pathogenicity of the isolates was confirmed by reproducing the symptoms on leaves, fruits, and twigs of the corresponding host plants. The biochemical tests indicated that the isolates belong to Pseudomonas syringae. However, isolates’ characterization showed variation in their phenotypic and molecular features. The presence of the syrB gene and ice nucleation activity grouped most of the isolates within pathovar syringae. The results of rep-PCR using the BOX primer revealed high genetic diversity of isolates. Multilocus sequence analysis (MLSA), using four housekeeping genes, showed that 27 isolates belong to the genomic species 1, P. syringae sensu stricto, corresponding to P. syringae phylogroup 2. However, isolates from the same phylogroup 2 did not form a monophyletic group. One strain isolated from apricot was most distinct and similar to members of genomic species 2, phylogroup 3. All tested isolates showed significant levels of resistance to copper sulfate and high level of sensitivity to streptomycin sulfate in vitro.

scholarly journals Establishment of an in vitro model for analyzing mitochondrial ultrastructure in PRKN-mutated patient iPSC-derived dopaminergic neurons

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Mutsumi Yokota ◽  
Soichiro Kakuta ◽  
Takahiro Shiga ◽  
Kei-ichi Ishikawa ◽  
Hideyuki Okano ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Dopaminergic Neurons ◽  
Structural Changes ◽  
In Vitro Model ◽  
Induced Pluripotent Stem Cell ◽  
Substantia Nigra Pars Compacta ◽  
Ultrastructural Changes ◽  
Mitochondrial Morphology ◽  
Vitro Model

AbstractMitochondrial structural changes are associated with the regulation of mitochondrial function, apoptosis, and neurodegenerative diseases. PRKN is known to be involved with various mechanisms of mitochondrial quality control including mitochondrial structural changes. Parkinson’s disease (PD) with PRKN mutations is characterized by the preferential degeneration of dopaminergic neurons in the substantia nigra pars compacta, which has been suggested to result from the accumulation of damaged mitochondria. However, ultrastructural changes of mitochondria specifically in dopaminergic neurons derived from iPSC have rarely been analyzed. The main reason for this would be that the dopaminergic neurons cannot be distinguished directly among a mixture of iPSC-derived differentiated cells under electron microscopy. To selectively label dopaminergic neurons and analyze mitochondrial morphology at the ultrastructural level, we generated control and PRKN-mutated patient tyrosine hydroxylase reporter (TH-GFP) induced pluripotent stem cell (iPSC) lines. Correlative light-electron microscopy analysis and live cell imaging of GFP-expressing dopaminergic neurons indicated that iPSC-derived dopaminergic neurons had smaller and less functional mitochondria than those in non-dopaminergic neurons. Furthermore, the formation of spheroid-shaped mitochondria, which was induced in control dopaminergic neurons by a mitochondrial uncoupler, was inhibited in the PRKN-mutated dopaminergic neurons. These results indicate that our established TH-GFP iPSC lines are useful for characterizing mitochondrial morphology, such as spheroid-shaped mitochondria, in dopaminergic neurons among a mixture of various cell types. Our in vitro model would provide insights into the vulnerability of dopaminergic neurons and the processes leading to the preferential loss of dopaminergic neurons in patients with PRKN mutations.

scholarly journals Antibiotic sensitivity and in vitro antimicrobial activity of plant extracts to pseudomonas fluorescens isolates collected from diseased fish

1970 ◽  
Vol 1 (4) ◽  
pp. 82-88 ◽  
Author(s):  
MJ Foysal ◽  
MM Rahman ◽  
M Alam
Keyword(s):  
Antibacterial Activity ◽  
Pseudomonas Fluorescens ◽  
Plant Extracts ◽  
Antibiotic Sensitivity ◽  
Diffusion Method ◽  
Biochemical Tests ◽  
Leaf Extracts ◽  
Sensitivity Pattern ◽  
Medicinal Plant Extracts

Studies were conducted to identify Pseudomonas fluorescens isolates from a collection of bacteria isolated from bacterial haemorrhagic septicaemia infected carp and catfish, evaluate their antibiotic sensitivity pattern and screen the antibacterial activity of some medicinal plant extracts against the isolates.. A total of 10 isolates were identified as P. fluorescens by morphological, physiological and biochemical tests. In vitro antibiotic sensitivity test of the P. fluorescens isolates were conducted by disc diffusion method for seven antibiotics where, all of the isolates were found to be sensitive only against streptomycin and gentamycin but, most of the isolates (80%) were found resistant to chloramphenicol (C). Moreover, eighty percent of the isolates showed resistance to multiple antibiotics. A total of 118 plant extracts were screened for their antibacterial activity against the P. fluorescens isolates where the isolates exhibited sensitivity to 30 samples. Leaf extracts of Tamarindus indicus, Terminalia chebula, Citrus aurantifolia, Eugenia caryophyllata and Spondias pinnata were found to inhibit the growth of all of the P. fluorescens isolates. DOI: http://dx.doi.org/10.3329/ijns.v1i4.9733 IJNS 2011 1(4): 82-88

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