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Potential of Piper Germplasm Against Pathogenic Bacteria: Tropical Bay Islands in India
Published in Megh R. Goyal, Durgesh Nandini Chauhan, Assessment of Medicinal Plants for Human Health, 2020
Chinthamani Jayavel, Ajit Arun Waman, Saravanan Kandasamy, Pooja Bohra
Antibacterial activity of the ethanolic extracts of Piper samples was determined using agar well diffusion method.16 For this, Mueller Hinton agar (HiMedia, Mumbai, India) plates were prepared and 50 µL of test culture was transferred aseptically to each plate and spread using a sterile L-rod followed by 30 min incubation. Using sterile cork borer (5.5 mm in diameter), four wells were made in each plate and 80 or 100 µL of prepared extract, as per the treatment, was inoculated in each well. Streptomycin (1 mg/mL) and chloramphenicol (1 mg/mL) were used as positive controls, whereas 80% ethanol was used as the negative control. Plates were then incubated at 37°C in an orbital shaking incubator (Remi, India). Each sample was maintained in triplicate to minimize the experimental error. Zone of inhibition in response to the challenging was measured after 24 h.
UV/V is Spectrophotometric Characterization of the Leaf Polyphenolics Content in Elaeocarpus tectorius and its Therapeutic Potential against Selected Urinary Tract Infection Pathogens
Published in Parimelazhagan Thangaraj, Phytomedicine, 2020
M. Ashwini Lydia, Suman Thamburaj, Gayathri Jagadeesan, Gayathri Nataraj, Kasipandi Muniyandi, Saikumar Sathyanarayanan, Parimelazhagan Thangaraj
The modified disk diffusion agar method (Olurinola 1996) was followed to verify the anti-microbial activity of E. tectorius leaf extracts against selected bacterial and fungal pathogens. The sterile swabs were used to swab the bacterial cultures from the broth onto the plates containing Mueller Hinton agar medium, similarly the fungal yeast pathogen ‘Candida’ were swabbed in the plate containing Sabouraud dextrose agar medium. Wells were made on the seeded plates with the help of a sterilized cork borer (6 mm Hi-Media). The collected different samples were further dissolved in 4% Dimethyl sulfoxide (DMSO). The diluted samples (100 μL of 10 mg/mL concentration) were dispended into the wells, and the plates were incubated aerobically at 37°C for 18–24 hours for bacterial pathogens and 28°C for 48 hours for fungal pathogens. In the same way, positive and negative control wells were made with Ampicillin-15 μg (bacteria), Miconazole-30 µg (Candida), and 4% DMSO, respectively. The entire microbial assay was carried out under strict aseptic conditions. The zones of inhibition (mm) of the samples were measured after incubation, and the activity index was also calculated. Triplicates were maintained for each pathogen, and the readings were taken in three different fixed directions, and the average values were recorded.
Antimicrobial Properties of Traditional Medicinal Plants: Status and Potential
Published in Megh R. Goyal, Durgesh Nandini Chauhan, Plant- and Marine-Based Phytochemicals for Human Health, 2018
V. Duraipandiyan, T. William Raja, Naif Abdullah Al-Dhabi, Ignacimuthu Savarimuthu
The agar plug diffusion method is similar to that of the disc diffusion method and includes making an agar culture of the microbial strain and applying tight streaks in a suitable culture medium. Microbial cells express their own molecules, which diffuse into the agar medium. After incubating, a sterile cork-borer is used to cut an agar plot, which is then deposited on another agar plate inoculated with test microorganisms. The substances then diffuse from the plug to the agar medium, forming zones of inhibition.
Repurposing of atorvastatin emulsomes as a topical antifungal agent
Published in Drug Delivery, 2022
Alaa S. Eita, Amna M. A. Makky, Asem Anter, Islam A. Khalil
Anticandidal activity using a well diffusion assay of free ATO suspension, ATO-EMLs, and blank EMLs formulae was conducted. First, SDA media was prepared and autoclaved. About 15–20 mL of the media was poured into sterile Petri dishes, spread homogeneously, and remained solidified for 30 min. Thereafter, 0.2 mL of inoculum (fungal strain in saline) was spread on an agar plate, and the excess was discarded via draining. The plates were then incubated at ambient temperature for 10 min. Subsequently, a ditch of 4 mm was shaped in each plate, using a sterile cork borer. Each well was filled with 50–100 μL of the diluted formulae. Commercial Fungistate 1% gel was used as positive control while saline was used as a negative control. The plates were incubated for 24 h at 37 °C. The zone of inhibition was measured in millimeters (Senyiğit et al., 2014). The assay was triplicated for confirmation.
Hydroxypropyl chitosan nail lacquer of ciclopirox-PLGA nanocapsules for augmented in vitro nail plate absorption and onychomycosis treatment
Published in Drug Delivery, 2022
Eman Yahya Gaballah, Thanaa Mohammed Borg, Elham Abdelmonem Mohamed
The selected NCs lacquer containing 1%w/v CIX, its blank and aqueous drug (1%w/v) solution containing 1%v/v tween 20 as a positive control were evaluated for their antifungal activity against Trichophyton rubrum applying the agar diffusion technique (Bseiso et al., 2016). Saline was included as a negative control. Sabouraud’s dextrose agar growth medium fortified with levofloxacin to guard against bacterial growth was added to a sterile petri dish (El-Emam et al., 2020). After agar solidification, 1 mL of the fungal suspension (105 CFU/mL) was added and the agar plate was moved roundly in a clockwise and anticlockwise directions. Four wells of 6 mm internal diameter were made in the agar using a sterile cork-borer. Each well was filled with 0.10 mL of the above-mentioned tested samples. The plates were then incubated at 28° C for three days to enhance the fungal growth (Bseiso et al., 2016). The antifungal activity was assessed by measuring the diameter (mm) of the formed inhibition zones surrounding the formulations using a scale.
Single-cell genomics for resolution of conserved bacterial genes and mobile genetic elements of the human intestinal microbiota using flow cytometry
Published in Gut Microbes, 2022
Dylan Lawrence, Danielle E. Campbell, Lawrence A. Schriefer, Rachel Rodgers, Forrest C. Walker, Marissa Turkin, Lindsay Droit, Miles Parkes, Scott A. Handley, Megan T. Baldridge
Human stool samples were obtained from a cohort of longitudinally sampled inflammatory bowel disease patients and their healthy household controls, collected from Addenbrooke’s Hospital, University of Cambridge, UK, and described previously.42 Patients 1, 4, and 6 were individuals with inflammatory bowel diseases: respectively, a female with Ulcerative Colitis, a male with Crohn’s disease, and another male with Crohn’s disease. Patients 2, 3, and 5 were household controls, and were, respectively, a healthy male, a healthy female paired with Patient 6, and a healthy female paired with Patient 1. Stool samples were removed from −80°C storage and immediately placed on dry ice. Samples were then cored using a small cork borer (Humboldt Manufacturing Company H9661). Core samples were transferred to 2 mL Eppendorf tubes containing homogenization beads. After homogenization, DNA for community profiling was extracted from these samples by phenol:chloroform extraction and cleaned with the DNeasy Blood and Tissue Kit (Qiagen 69504). DNA was prepared as outlined in the “16S rRNA gene sequencing and analysis” section below.