scholarly journals Antibacterial and Antibiofilm Activities of Crude Extract of Lasiodiplodia pseudotheobromae IBRL OS-64 against Foodborne Bacterium, Yersinia enterocolitica

2020 ◽  
pp. 87-102
Author(s):  
M. M. J. Taufiq ◽  
I. Darah
Keyword(s):  
Ethyl Acetate ◽  
Yersinia Enterocolitica ◽  
Foodborne Pathogens ◽  
Crude Extract ◽  
Extracellular Polysaccharide ◽  
Antibiofilm Activity ◽  
Bacterial Cells ◽  
Transmission Electron ◽  
Inhibition Concentration ◽  
Initial Biofilm

The antibacterial and antibiofilm activities of crude extract of Lasiodiplodia pseudotheobromae IBRL OS-64 was studied and tested against a foodborne pathogenic bacterium, Yersinia enterocolitica. The ethyl acetate extract exhibited favorable antibacterial activity with the zone of inhibition was 20.3±0.6 mm compared to dichloromethane (15.0±0.3 mm) and butanol (9.0±0.3 mm) extracts. Minimum inhibition concentration (MIC) and minimum bactericidal concentration (MBC) values of the extract were 125 and 250 µg/mL, respectively. Structural degeneration studies through scanning electron microscopy (SEM) and transmission electron microscope (TEM) micrographs exhibited major abnormalities that occurred on the bacterial cells after exposure to the extract were complete alterations in their morphology and collapsed of the cells beyond repair. The findings showed that the extract possesses antibiofilm activity against the initial and preformed biofilm of Y. enterocolitica with the highest inhibition value of 69.12% and 58.70%, respectively. The results also revealed the initial biofilm was more susceptible to the extract as compared to pre-formed biofilm. The light microscopy (LM) and SEM photomicrographs proved that the fungal extract significantly eliminates extracellular polysaccharide (EPS) matrices and hinder the attachment of the bacterial cells for biofilm formation. Therefore, the current study suggested the ethyl acetate crude extract from an endophytic fungus, L. pseudotheobromae IBRL OS-64 may be an effective antibacterial and anti-biofilm agent to treat foodborne pathogens.

scholarly journals Optimization of an Extended Microplate Assay for Generating Listeria monocytogenes, E. coli O157:H7, and Salmonella Biofilms and Enzymatic Recovery for Enumeration

2019 ◽  
Author(s):  
Manish Aryal ◽  
Preetty Pranatharthiharan ◽  
Peter M. Muriana
Keyword(s):  
Foodborne Pathogens ◽  
Food Processing ◽  
Hydrolytic Enzymes ◽  
Extracellular Polysaccharide ◽  
Fluorescence Assay ◽  
Bacterial Cells ◽  
Microplate Assay ◽  
Planktonic Cells ◽  
E Coli ◽  
Microplate Fluorescence Assay

Biofilms enable the persistence of pathogens in food processing environments. In order to inactivate these microorganisms, sanitizing agents are needed that are effective against pathogens entrapped in biofilms which are more difficult to inactivate than planktonic cells that are displaced and found on equipment surfaces. We examined conditions to develop, analyze, and enumerate robust biofilms of 3 different foodborne pathogens assisted by fluorescence adherence assay and enzymatic detachment. We compared 3 different isomeric forms of fluorescent substrates that are readily taken up by bacterial cells based on carboxy-fluorescein diacetate (5-CFDA, 5,6-CFDA, 5,6-CFDA,SE). Biofilm-forming strains of L. monocytogenes, E.coli O157:H7, and Salmonella serovars were detected using the chosen substrate in a microplate fluorescence assay define previously for use with Listeria. Adherence levels were determined by differences in relative fluorescence units (RFU). Multiple hydrolytic enzymes were examined for each representative pathogen for the most suitable enzyme for detachment and enumeration to confirm data obtained by fluorescence assay. Cultures were grown overnight in microplates, incubated, washed and replenished with fresh sterile growth medium; this cycle was repeated 7 times before used as ‘biofilms’. All treatments were performed in triplicate and compared by one-way analysis of variance (ANOVA) to determine significant differences (p < 0.05). Analysis of 7-day biofilms by SEM indicated possible extracellular polysaccharide involvement with Salmonella and E. coli.

Phytochemical screening for identification of bioactive compounds of Lantana camara Linn. (Verbenaceae) responsible for larvicidal action against Aedes triseriatus Say, 1823 (Diptera : Culicidae)

ENTOMON ◽  
2019 ◽  
Vol 44 (2) ◽  
pp. 117-126
Author(s):  
Rhitayu Chakraborti ◽  
Probir Kumar Bandyopadhyay
Keyword(s):  
Ethyl Acetate ◽  
Bioactive Compounds ◽  
Active Ingredient ◽  
Crude Extract ◽  
Ethyl Acetate Extract ◽  
Lantana Camara ◽  
Mosquito Larvae ◽  
Aedes Triseriatus ◽  
Phytochemical Screening ◽  
Lc50 Value

Study to assess the larvicidal property of Lantana camara leaves against Aedes triseriatus larvae found that the ethyl acetate extract had profound larvicidal action with the crude extract having a LC50 value of 409.831ppm. GC-MS analysis of the ethyl acetate extract confirmed the presence of twenty-one compounds out of which beta-caryophyllene covered the highest percentage of the chromatogram area. Further tests with beta-caryophyllene against the mosquito larvae proved it to be the active ingredient of L. Camara with a LC50 value of 104.243ppm.

scholarly journals CdSe QD Biosynthesis in Yeast Using Tryptone-Enriched Media and Their Conjugation with a Peptide Hecate for Bacterial Detection and Killing

Nanomaterials ◽  
2019 ◽  
Vol 9 (10) ◽  
pp. 1463 ◽  
Author(s):  
Vishma Pratap Sur ◽  
Marketa Kominkova ◽  
Zaneta Buchtova ◽  
Kristyna Dolezelikova ◽  
Ondrej Zitka ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Synthesis Process ◽  
Synthesis Methods ◽  
Bacterial Cells ◽  
Cdse Qds ◽  
Yeast Saccharomyces Cerevisiae ◽  
Shape Distribution ◽  
Transmission Electron ◽  
A Cell ◽  
Physical And Chemical

The physical and chemical synthesis methods of quantum dots (QDs) are generally unfavorable for biological applications. To overcome this limitation, the development of a novel “green” route to produce highly-fluorescent CdSe QDs constitutes a promising substitute approach. In the present work, CdSe QDs were biosynthesized in yeast Saccharomyces cerevisiae using a novel method, where we showed for the first time that the concentration of tryptone highly affects the synthesis process. The optimum concentration of tryptone was found to be 25 g/L for the highest yield. Different methods were used to optimize the QD extraction from yeast, and the best method was found to be by denaturation at 80 °C along with an ultrasound needle. Multiple physical characterizations including transmission electron microscopy (TEM), dynamic light scattering (DLS), energy-dispersive X-ray spectroscopy (EDX), and spectrophotometry confirmed the optical features size and shape distribution of the QDs. We showed that the novel conjugate of the CdSe QDs and a cell-penetrating peptide (hecate) can detect bacterial cells very efficiently under a fluorescent microscope. The conjugate also showed strong antibacterial activity against vancomycin-resistant Staphylococcus aureus (VRSA), methicillin-resistant Staphylococcus aureus (MRSA), and Escherichia coli, which may help us to cope with the problem of rising antibiotic resistance.

scholarly journals Zinc Oxide and Silver Nanoparticle Effects on Intestinal Bacteria

Materials ◽  
2021 ◽  
Vol 14 (10) ◽  
pp. 2489
Author(s):  
Ami Yoo ◽  
Mengshi Lin ◽  
Azlin Mustapha
Keyword(s):  
Electron Microscopy ◽  
Zinc Oxide ◽  
Morphological Changes ◽  
Intestinal Bacteria ◽  
Bacterial Cells ◽  
Ag Nps ◽  
Zno Nps ◽  
Bifidobacterium Animalis ◽  
Transmission Electron

The application of nanoparticles (NPs) for food safety is increasingly being explored. Zinc oxide (ZnO) and silver (Ag) NPs are inorganic chemicals with antimicrobial and bioactive characteristics and have been widely used in the food industry. However, not much is known about the behavior of these NPs upon ingestion and whether they inhibit natural gut microflora. The objective of this study was to investigate the effects of ZnO and Ag NPs on the intestinal bacteria, namely Escherichia coli, Lactobacillus acidophilus, and Bifidobacterium animalis. Cells were inoculated into tryptic soy broth or Lactobacilli MRS broth containing 1% of NP-free solution, 0, 12, 16, 20 mM of ZnO NPs or 0, 1.8, 2.7, 4.6 mM Ag NPs, and incubated at 37 °C for 24 h. The presence and characterization of the NPs on bacterial cells were investigated by scanning electron microscopy (SEM), transmission electron microscopy (TEM), and energy-dispersive X-ray spectroscopy (EDS). Membrane leakage and cell viability were assessed using a UV-visible spectrophotometer and confocal electron microscope, respectively. Numbers of treated cells were within 1 log CFU/mL less than those of the controls for up to 12 h of incubation. Cellular morphological changes were observed, but many cells remained in normal shapes. Only a small amount of internal cellular contents was leaked due to the NP treatments, and more live than dead cells were observed after exposure to the NPs. Based on these results, we conclude that ZnO and Ag NPs have mild inhibitory effects on intestinal bacteria.

HSCCC Separation of Three Main Compounds from the Crude Extract of Dracocephalum Tanguticum by Using Dimethyl Sulfoxide as Cosolvent

2020 ◽  
Author(s):  
Xue Yang ◽  
Yongling Liu ◽  
Tao Chen ◽  
Nana Wang ◽  
Hongmei Li ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Ethyl Acetate ◽  
Dimethyl Sulfoxide ◽  
Efficient Method ◽  
Crude Extract ◽  
High Speed ◽  
Glacial Acetic Acid ◽  
Butyl Alcohol ◽  
Preparative Hplc ◽  
Counter Current

Abstract Separation of natural compounds directly from the crude extract is a challenging work for traditional column chromatography. In the present study, an efficient method for separation of three main compounds from the crude extract of Dracocephalum tanguticum has been successfully established by high-speed counter-current chromatography (HSCCC). The crude extract was directly introduced into HSCCC by using dimethyl sulfoxide as cosolvent. Ethyl acetate/n-butyl alcohol/0.3% glacial acetic acid (4: 1: 5, v/v) system was used and three target compounds with purity higher than 80% were obtained. Preparative HPLC was used for further purification and three target compounds with purity higher than 98% were obtained. The compounds were identified as chlorogenic acid, pedaliin and pedaliin-6″-acetate.

scholarly journals Isolation and characterization of pathogenic Escherichia coli bacteriophages from chicken and beef offal

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Celosia Lukman ◽  
Christopher Yonathan ◽  
Stella Magdalena ◽  
Diana Elizabeth Waturangi
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogens ◽  
High Efficiency ◽  
Multiplicity Of Infection ◽  
Isolation And Characterization ◽  
Transmission Electron ◽  
Pathogenic Escherichia Coli ◽  
Family Based

Abstract Objective This study was conducted to isolate and characterize lytic bacteriophages for pathogenic Escherichia coli from chicken and beef offal, and analyze their capability as biocontrol for several foodborne pathogens. Methods done in this research are bacteriophage isolation, purification, titer determination, application, determination of host range and minimum multiplicity of infection (miMOI), and bacteriophage morphology. Results Six bacteriophages successfully isolated from chicken and beef offal using EPEC and EHEC as host strain. Bacteriophage titers observed between 109 and 1010 PFU mL−1. CS EPEC and BL EHEC bacteriophage showed high efficiency in reduction of EPEC or EHEC contamination in meat about 99.20% and 99.04%. The lowest miMOI was 0.01 showed by CS EPEC bacteriophage. CI EPEC and BL EPEC bacteriophage suspected as Myoviridae family based on its micrograph from Transmission Electron Microscopy (TEM). Refers to their activity, bacteriophages isolated in this study have a great potential to be used as biocontrol against several foodborne pathogens.

scholarly journals Microbial Photoinactivation by Visible Light Results in Limited Loss of Membrane Integrity

Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 341
Author(s):  
Katharina Hoenes ◽  
Richard Bauer ◽  
Barbara Spellerberg ◽  
Martin Hessling
Keyword(s):  
Visible Light ◽  
Pseudomonas Fluorescens ◽  
Bacterial Cell ◽  
Bacterial Species ◽  
Membrane Integrity ◽  
Cell Lysis ◽  
Microbial Inactivation ◽  
Bacterial Cells ◽  
Resistance Development ◽  
Transmission Electron

Interest in visible light irradiation as a microbial inactivation method has widely increased due to multiple possible applications. Resistance development is considered unlikely, because of the multi-target mechanism, based on the induction of reactive oxygen species by wavelength specific photosensitizers. However, the affected targets are still not completely identified. We investigated membrane integrity with the fluorescence staining kit LIVE/DEAD® BacLight™ on a Gram positive and a Gram negative bacterial species, irradiating Staphylococcus carnosus and Pseudomonas fluorescens with 405 nm and 450 nm. To exclude the generation of viable but nonculturable (VBNC) bacterial cells, we applied an ATP test, measuring the loss of vitality. Pronounced uptake of propidium iodide was only observed in Pseudomonas fluorescens at 405 nm. Transmission electron micrographs revealed no obvious differences between irradiated samples and controls, especially no indication of an increased bacterial cell lysis could be observed. Based on our results and previous literature, we suggest that visible light photoinactivation does not lead to rapid bacterial cell lysis or disruption. However, functional loss of membrane integrity due to depolarization or inactivation of membrane proteins may occur. Decomposition of the bacterial envelope following cell death might be responsible for observations of intracellular component leakage.

Pseudomonas aeruginosa Oligoribonuclease Controls Susceptibility to Polymyxin B by Regulating Pel Exopolysaccharide Production

2022 ◽  
Author(s):  
Baopeng Yang ◽  
Yujun Jiang ◽  
Yongxin Jin ◽  
Fang Bai ◽  
Zhihui Cheng ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Bacterial Resistance ◽  
Defense Mechanism ◽  
Extracellular Polysaccharide ◽  
Opportunistic Pathogen ◽  
Polymyxin B ◽  
Guanosine Monophosphate ◽  
Extracellular Dna ◽  
Bacterial Cells ◽  
Bacterial Survival

Polymyxins are considered as the last resort antibiotics to treat infections caused by multidrug-resistant Gram negative pathogens. Pseudomonas aeruginosa is an opportunistic pathogen that causes various infections in humans. Proteins involved in lipopolysaccharide modification and maintaining inner and outer membrane integrities have been found to contribute to the bacterial resistance to polymyxins. Oligoribonuclease (Orn) is an exonuclease that regulates the homeostasis of intracellular (3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP), thereby regulating the production of extracellular polysaccharide in P. aeruginosa . Previously, we demonstrated that Orn affects the bacterial resistance to fluoroquinolone, β-lactam and aminoglycoside antibiotics. In this study, we found that mutation of orn increased the bacterial survival following polymyxin B treatment in a wild type P. aeruginosa strain PA14. Overexpression of c-di-GMP degradation enzymes in the orn mutant reduced the bacterial survival. By using a fluorescence labeled polymyxin B, we found that mutation of orn increased the bacterial surface bound polymyxin B. Deletion of the Pel synthesis genes or treatment with a Pel hydrolase reduced the surface bound polymyxin B and bacterial survival. We further demonstrated that Pel binds to extracellular DNA (eDNA), which traps polymyxin B and thus protects the bacterial cells. Collectively, our results revealed a novel defense mechanism against polymyxin in P. aeruginosa .

scholarly journals Adhesive properties of an outer structure of Clostridium perfringens type A isolated from piglets with with catarrhal enteritis

1999 ◽  
Vol 30 (3) ◽  
pp. 242-248 ◽  
Author(s):  
Elizabeth Pelosi Teixeira ◽  
Marlene Braide Serafim ◽  
Maria Alice Cruz Höfling ◽  
Aureo T. Yamada ◽  
Antonio Fernando Pestana de Castro
Keyword(s):  
Clostridium Perfringens ◽  
Type A ◽  
Negative Control ◽  
Bacterial Cells ◽  
Intestinal Epithelial ◽  
Adhesive Properties ◽  
Ileal Loop ◽  
Fibrillar Material ◽  
Gram Staining ◽  
Transmission Electron

One strain (S32) of Clostridium perfringens type A was isolated from a case of catarrhal enteritis of piglets. This strain was able to adhere to HeLa cells showing an adherence index (AI) of 25.15 ± 1.26 (mean ± 1 standard error of the mean). Treatment of the bacterial cells with trypsin (0.25mg/ml) decreased in 70%-80% the AI and metaperiodate (10mg/ml) abolished completely the adherence, suggesting that the structure responsible for this phenomenon was probably a glycoprotein. Heating of bacterial suspensions (100ºC/5 min) before carrying out the adhesion test decreased the AI rendering it equal to the negative controls. Rabbit homologous S32 antiserum inhibited the adherence up to dilutions of 1: 640, at least. The piglet ileal loop assay, carried out with strains S32 and Jab-1 (negative control) demonstrated that the strain S32 was able to adhere to the intestinal epithelial cells when examined after Gram staining. Transmission electron microcopy (TEM) demonstrated that S32 strain displayed a loose fibrillar material not seen with Jab-1. Stabilization of the bacterial cells with homologous antiserum of strain S32, followed by staining with rhuteniun red, revealed loose long fibrillar material on the outer surface of the cells, that sometimes could be seen spreading out from the cells and linking bacterial cells. The question whether this structure might be an adhesin for this strain of Cl. perfringes type A, perhaps playing a role in the pathogenesis of the catarrhal enteritis of piglets, is dependent on further studies.

scholarly journals Chromate Reduction in Serratia marcescens Isolated from Tannery Effluent and Potential Application for Bioremediation of Chromate Pollution

2002 ◽  
Vol 2 ◽  
pp. 972-977 ◽  
Author(s):  
M.A. Mondaca ◽  
V. Campos ◽  
R. Moraga ◽  
C.A. Zaror
Keyword(s):  
Natural Waters ◽  
Waste Treatment ◽  
Environmental Concern ◽  
Electron Microscopic Examination ◽  
Tannery Effluent ◽  
Chromate Reduction ◽  
Bacterial Cells ◽  
Electron Microscopic ◽  
Treatment Processes ◽  
Transmission Electron

Pollution of aquatic systems by heavy metals has resulted in increasing environmental concern because they cannot be biodegraded. One metal that gives reason for concern due to its toxicity is chromium. Cr(VI) and Cr(III) are the principal forms of chromium found in natural waters. A chromate-resistant strain of the bacterium S. marcescens was isolated from tannery effluent. The strain was able to reduce Cr(VI) to Cr(III), and about 80% of chromate was removed from the medium. The reduction seems to occur on the cell surface. Transmission electron microscopic examination of cells revealed that particles were deposited on the outside of bacterial cells. A stable biofilm was formed in less than 10 h, reaching around 1010cfu attached per milligram of activated carbon. These findings demonstrate that immobilizedS. marcescensmight be used in industrial waste treatment processes.

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