American Type Culture Collection Research Articles

Green synthesized silver nanoparticles using carrot extract exhibited strong antibacterial activity against multidrug resistant bacteria

Antimicrobial resistance is a worldwide problem. Pathogenic microorganisms develop antibiotic resistance as a result of exposure to inappropriate quantity of antibiotics and accumulation of mutations. Bio-friendly nanomaterial scan be developed as antimicrobial agents. Present study aimed to investigate the activity of biosynthesized silver nanoparticles (AgNPs) obtained from carrot extract of three different regions namely Gilgit, Haripur and Sheikhupura from Pakistan, having significant difference in climate and altitude. UV–visible spectroscopy, Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analysis were carried out for the morphological characterization of synthesized nanoparticles. Analysis of XRD revealed crystalline nature of AgNPs and average size calculated was about 22, 17 and 10 nm from Gilgit, Haripur and Sheikhupura samples respectively. The antibacterial efficacy of AgNPs was carried out against 7 American type culture collection strains of pathogenic bacteria and three macrolide resistant clinical isolates with promising results. Moreover, antioxidant activity indicated maximum DPPH radical scavenging effects as 47 % using a concentration of 250 μg/mL. Hemolysis assays indicated biocompatibility of AgNPs at lower concentration like 7 μg/mL and 15 μg/mL. Comparative analysis of bioactivity from different sites of sampling had indicated minute differences which may be due to the change in altitude, soil texture and other local environmental changes.

Chemical Compositions and Antimicrobial Activity of Essential Oils From the Leaves of 4 Vietnamese Zingiberaceae Species

Essential oils of the leaves of 4 Vietnamese Zingiberaceae species were first obtained by hydro-distilled, and their chemical compositions were identified by gas chromatography-flame ionization detection/mass spectrometry . β-Pinene (21.7%), sabinene (12.8%), and α-pinene (8.0%) were the main compounds in Wurfbainia tenella leaf oil. The essential oil of Hedydium villosum var. tenuifolium leaf was dominated by β-pinene (23.7%), β-caryophyllene (21.6%), and 1,8-cineole (14.0%). β-Pinene (27.4%), α-pinene (21.2%), limonene (12.1%), and myrcene (8.6%) were the characteristic components of Meistera sudae leaf oil, and β-pinene (32.8%) and ( E)-methyl cinnamate (15.8%) of Alpinia hongiaoensis leaf oil. The 4 oil samples exhibited antimicrobial activity against Bacillus subtilis (American Type Culture Collection [ATCC] 27212), Staphylococcus aureus (ATCC 12222), Escherichia coli (ATCC 8739), Pseudomonas aeruginosa (ATCC 25923), Aspergillus niger (ATCC 9763), Fusarium oxysporum (ATCC 48112), Candida albicans (ATCC 10231), and Saccharomyces cerevisiae (ATCC 2601) at different levels. Especially, the leaf oil of M sudae showed strong activity against B subtilis, S aureus, and S cerevisiae with a minimum inhibitory concentration (MIC) value of 25.0 µg/mL.

Draft Genome Sequence of a Xylella fastidiosa Strain Causing Bacterial Leaf Scorch of American Elm in Washington, DC.

Here, we report the draft genome sequence of Xylella fastidiosa strain ATCC 35873, which was obtained from the American Type Culture Collection and was originally isolated from a symptomatic American elm tree grown in Washington, DC. The ATCC 35873 genome contains 2,454,216 bp and has a GC content of 51.68%.

Effect of antioxidant lycopene on human osteoblasts.

The aim of this in vitro study is to evaluate the effect of antioxidant lycopene on human osteoblasts. The human osteoblast cell line (CRL-11372) was obtained from the American Type Culture Collection (ATCC Manassas, Va) and grown in Dulbecco's Modified Eagle's medium (DMEM) supplemented with 10% fetal calf serum (FCS), penicillin (100 U/ml), and streptomycin (100mg/ ml) at 37°C in a humidified atmosphere of 5% CO2 and 95% air. The effective dose of lycopene was determined by MTT assay and a real-time cell analysis (RTCA) system. Proliferative effects were analyzed by in vitro wound healing model. Gene expressions of type 1 collagen (COL1A1), osteocalcin (OCN), and growth differentiation factor-5 (GDF-5) were measured by quantitative real-time polymerase chain reaction (qRT-PCR) at 72h. Statistical differences between test groups were analyzed with a one-way ANOVA test. MTT assay showed that the doses between 10-5 and 1µmol of lycopene had dose-dependent proliferative effects. The doses between 10-5 and 10-1µmol were most effective at 72h. Lycopene accelerates the healing rate by increasing osteoblast proliferation. Results suggested that lycopene had proliferative effects on human osteoblasts, which may help to increase bone regeneration, and thus, it can be useful in tissue engineering procedures. By the help of antioxidants like lycopene capacity, velocity and quality of new bone forming may be increased in periodontal and dental implant treatments.

Yogurt fortified with GABA-producing strain and Ganoderma lucidum industrial waste

This study aimed to produce yogurt with Ganoderma lucidum residues and selected probiotic bacteria. To select the most potent GABA-producing strain (GABA – γ-aminobutyric acid), nine probiotic bacteria were subjected to the glutamate decarboxylase (GAD) activity assay. Limosilactobacillus reuteri DSM 17938 was selected and used in preparing fresh yogurts, with and without supplementation of Ganoderma residues obtained after water extraction [GW (Ganoderma waste) 2% (w/v)]. A decrease in pH during fermentation and the occurrence of syneresis were investigated. Lactic acid bacteria (LAB) viability and anti-Escherichia activity were estimated. Further, the cytotoxic effect of yogurt extracts on the human colon cancer cell line (HCT116) was surveyed. Besides functionality, the sensory attributes were evaluated. The pH values did not significantly change with the GW addition while increasing the LAB counts [up to 9.76 ± 0.11 log colony-forming units (CFU) mL<sup>–1</sup>] and the cytotoxic effect on HCT116 cells. Yogurt produced with selected bacteria and GW had the most valuable anti-coli effect against E. coli 0157: H7 and E. coli ATCC 35218 (American Type Culture Collection, Rockville, Maryland) (4.81 ± 0.62 and 5.64 ± 0.29 CFU mL<sup>–1</sup>, respectively). Although the added GW increased the yogurt functionality, it had a slightly negative effect on the taste and texture of the partially modified recipe. Yogurts fortified with GABA-producing strain and GW could potentially reduce relapse rates of depressive disorders.

In vitro biodegradation, cytotoxicity, and biocompatibility of polylactic acid/napier cellulose nanofiber scaffold composites

This study aimed to evaluate the bioactivities and biocompatibilities of porous polylactic acid (PLA) reinforced with cellulose nanofiber (CNF) scaffolds. The in vitro degradation behaviors of the porous PLA/CNF scaffolds were systematically measured for up to 8 weeks in a phosphate-buffered saline medium at 37 °C. The reinforcement of CNF resisted the biodegradation of the scaffolds. The in vitro cytotoxicity and biocompatibility of the scaffolds were determined using the Beas2B American Type Culture Collection cells. The 3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide cytotoxicity and proliferation tests showed that the scaffolds were non-toxic, and epithelial cells grew well on the scaffold after 7 days of culture, whereas the percentage of cell proliferation on the PLA/CNF15 scaffold was the largest, 130 %. A scratch wound-healing assay was performed to evaluate the suitability of the scaffolds for cell migration. The results demonstrated that the scaffolds exhibited good cell migration towards nearly complete wound closure.

Fast and New Microbiological Method for Evaluating the Potency of Marbofloxacin-Based Tablets.

Marbofloxacin (MAR) is an antimicrobial belonging to the fluoroquinolone class and is marketed in the form of tablets.There is no microbiological monograph in official compendia to evaluate the potency of the final product. Furthermore, the literature is scarce in this context. The objective is to develop and validate a microbiological method by turbidimetry to evaluate the potency of MAR-based tablets, based on the principles of green analytical chemistry. Purified water-ethanol (9 + 1, v/v) was used as diluent to prepare the MAR solutions at concentrations of 0.25, 0.8, and 2.56 µg/mL, brain heart infusion (BHI) broth as culture media, E. coli American Type Culture Collection (ATCC) 25922 at 10% and incubation in a shaker at 37°C for 4 h were used in the method. The method was: linear in the range of 0.25 to 2.56 µg/mL; selective against the sample adjuvants; precise (intra-day RSD 2.26%, inter-day RSD 3.49%, and inter-analyst RSD 3.59%); accurate with a recovery of 100.20%; and robust against changes in culture medium volume in the tube, shaker rotation, and incubation temperature in the shaker. The potency of MAR tablets was 96.98% using the proposed method. The turbidimetric method developed is a new, fast, and optimized option to the routine QC of MAR in tablets, since it allows the evaluation of the drug's potency in the final product and can be used to complement the results of the physicochemical analysis, in addition to being a green and sustainable alternative. The work shows an ecological and green alternative to the routine microbiological pharmaceutical analysis of MAR tablets.

“On Water” Effect for Green Click Reaction: Spontaneous Polymerization of Activated Alkyne with “Inert” Aromatic Amine in Aqueous Media

“On Water” Effect for Green Click Reaction: Spontaneous Polymerization of Activated Alkyne with “Inert” Aromatic Amine in Aqueous Media

A Genome Sequence Assembly of the Phototactic and Optogenetic Model Fungus Blastocladiella emersonii Reveals a Diversified Nucleotide-Cyclase Repertoire.

The chytrid fungus Blastocladiella emersonii produces spores with swimming tails (zoospores); these cells can sense and swim toward light. Interest in this species stems from ongoing efforts to develop B. emersonii as a model for understanding the evolution of phototaxis and the molecular cell biology of the associated optogenetic circuits. Here, we report a highly contiguous genome assembly and gene annotation of the B. emersonii American Type Culture Collection 22665 strain. We integrate a PacBio long-read library with an Illumina paired-end genomic sequence survey leading to an assembly of 21 contigs totaling 34.27 Mb. Using these data, we assess the diversity of sensory system encoding genes. These analyses identify a rich complement of G-protein-coupled receptors, ion transporters, and nucleotide cyclases, all of which have been diversified by domain recombination and tandem duplication. In many cases, these domain combinations have led to the fusion of a protein domain to a transmembrane domain, tying a putative signaling function to the cell membrane. This pattern is consistent with the diversification of the B. emersonii sensory-signaling systems, which likely plays a varied role in the complex life cycle of this fungus.

Étude in vitro de la cinétique d’élution et de l’activité biologique de la pipéracilline/tazobactam et de la gentamicine dans du ciment osseux acrylique

BackgroundAntibiotics differ in their elution characteristics from bone cement. But no such data is available on piperacillin and tazobactam. Therefore, we performed an in vitro observational study to examine: (1) in-vitro elution characteristics of piperacillin and tazobactam from bone cement, (2) their biological activity using minimum inhibitory concentration and (3) elution characteristics and biological activity when combined with gentamicin in bone cement. HypothesisThe null hypothesis was that piperacillin and tazobactam after elution from bone cement can achieve concentrations higher than minimum inhibitory concentration. Material and methodsForty milligrams bone cement was mixed with the following combination of antibiotics: without any antibiotic (Sample A, Control), 4g/0.50g piperacillin/tazobactam (Sample B), 6g/0.75g piperacillin/tazobactam (Sample C), 8g/1.0g piperacillin/tazobactam (Sample D) and 4g/0.50g piperacillin/tazobactam and 400mg gentamicin (Sample E). Samples were analysed on reverse-phase ultra-high-performance liquid chromatography. Antibacterial activity in the elute were tested against standard ATCC (American Type Culture Collection) strains. ResultsDetectable drug elution for piperacillin and tazobactam was seen till 21 days. Peak drug levels for all formulations were seen at 48hours (140.8 and 297.5μg/ml for Samples B of piperacillin and tazobactam respectively). About 0.83–1.24% of piperacillin and 23.17-29.17% of tazobactam were released from the samples. Gentamicin improved elution of piperacillin and tazobactam: 140.8 vs 919.9μg/ml (p = 0.000) for Samples B and E of piperacillin respectively and 297.5 and 1138.4μg/ml (p = 0.001) for Samples B and E of tazobactam respectively at 2 days. Sample E showed complete inhibition of tested microorganisms, while B sample was microbiologically less active compared to E on day 5. ConclusionsPiperacillin and tazobactam eluted successfully from bone cement and also retained antimicrobial activity after elution. Maximum elution was seen up to day 2 after which it reduced drastically. Antimicrobial action was seen up to 7 days. Level of evidenceIII; comparative study.

Influence of melatonin associated with the Bio-Gide® membrane on osteoblast activity: an in vitro Study.

Melatonin (MLT) is a hormone responsible for regulating several physiological processes. It has been shown that MLT can be an important mediator in bone formation and stimulation, promoting osteoblast differentiation. In clinical practice, in tissue regeneration procedures, it is necessary to use membranes or barriers, associated with biomaterials, or not. The aim of this in vitro study was to assess the effect of melatonin on the activity of osteoblastic cells, associated, or not, with a resorbable collagen membrane (Bio-Gideä). For this, mice-derived pre-osteoblastic cells MC3T3 obtained from the ATCC (American Type Culture Collection) were used. Cultured cells were subject to the following treatments: MLT with a concentration of 1mM, a Bio-Gideä membrane and a membrane associated with MLT (Bio-Gideä + MLT). Proliferation and cell viability assays and protein lysate (ELISA test) quantification for the BMP-2 protein were carried out, in periods of 72 hours, 7 days and 10 days. After analyzing the data (one-way ANOVA, alpha=5%) it was observed that when MLT was used in isolation, there was an increase in cell proliferation and viability in osteoblastic cells (p<0.05). But, when MLT was associated with resorbable membranes, there was an inverse behavior, both in terms of proliferation and viability (p<0.05). In the case of the ELISA test, no secretion of BMP-2 was detected in any of the analyzed groups. It is concluded that MLT has a stimulatory effect on osteoblasts, but, when associated with Bio-Gideä resorbable membranes, it does not show any viable action in osteoblastic cell stimulation.

Glycosylation of Ganoderic Acid F by Bacillus Glycosyltransferase

Ganoderma lucidum is a medicinal fungus and has been used for improvements of health or prevention of certain diseases in Asia for thousands of years. Despite numerous kinds of triterpenoids having been identified from G. lucidum, few natural Ganoderma triterpenoids exist in the form of glycosides (saponins). To expand the diversity of Ganoderma triterpenoids and find rare Ganoderma saponins, ganoderic acid F (GAF), a Ganoderma triterpenoid, was biotransformed by a glycosyltransferase (BsGT110) from Bacillus subtilis ATCC (American type culture collection) 6633. The results showed that BsGT110 catalyzed biotransformation of GAF to produce a metabolite, which was confirmed as a GAF glucoside by mass–mass spectroscopy. The GAF glucoside showed 89-fold higher aqueous solubility than that of GAF. The present study highlights the utility of BsGT110 in the production of novel Ganoderma triterpenoid saponins, and the newly identified and highly soluble GAF glucoside can be studied for its bioactivity in the future.

Establishment and characterization of a new mantle cell lymphoma cell line with a NOTCH2 mutation, Arbo.

Cell lines represent an essential tool used in preclinical research. Most hematologic malignancies have a wide array of cell lines representing their respective molecular and pathologic spectra. In mantle cell lymphoma (MCL), cell lines become specifically valuable in view of the heterogeneity of this disease. Unfortunately, the number of MCL cell lines that are available for the research community remains small, with only nine cell lines available for purchase through the American Type Culture Collection (ATCC). We have established a novel blastoid MCL cell line, isolated from the malignant pleural effusion of a 69-year-old male with refractory MCL. Arbo was fully characterized with cytogenetics, immunophenotyping, whole exome sequencing and drug sensitivity assays. One of the most notable mutations identified in Arbo (but not in normal tissue) was the missense mutation NOTCH2 R2400*, which has been proposed as a clinically significant mutation in MCL seen in 5% of cases. NOTCH2 R2400* results in a truncated Notch2 protein, leading to a more stable and active protein. Using pharmacologic inhibition of Notch2, we showed a dependence of Arbo on NOTCH2 signaling, as well as a link between CD23 expression on Arbo and NOTCH2 activity. Arbo represents a NOTCH2 mutated model that is useful in MCL as well as other lymphomas with such mutation. We plan to deposit Arbo at the ATCC to be available for the research community.

P042 Identification, clinical profile, antifungal susceptibility pattern of candida auris from a tertiary care center in india

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PMObjectivesTo identify the phenotypic characteristics of Candida auris.To analyze the clinical profile of Candida auris infection.To describe the antifungal susceptibility pattern of Candida auris.MethodsThe study was conducted in the Department of Microbiology in Mycology division at Sri Ramachandra Institute of Higher Education and Research from December 2019 to November 2021. The study protocol was approved by Institutional Ethics Committee. Candida species isolated from various specimens sent to the laboratory were identified by Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight mass spectrometry (MALDI-TOF). The growth characteristics of C. auris were investigated on various media including Selective Auris Medium (SAM), HiChrome agar Candida and Tetrazolium reduction agar.Antifungal susceptibility testing was performed by using the Clinical and Laboratory Standards Institute broth microdilution method M27-A3. Antifungals tested were fluconazole, itraconazole, voriconazole, posaconazole, micafungin, anidulafungin, caspofungin and amphotericin B. Candida albicans American Type Culture Collection (ATCC) 22 019 was used as quality control strains.Data were collected for demographics, risk factors for candidemia, treatment, and outcome from the respective wards and ICUs.ResultsA total of 37 C. auris isolates were collected. Both adult and pediatric cases were included. The majority (23.3%) of the C. auris cases were seen in the age group of 55-64. Median age was 54 years for the adults. Among the 7 children, 6 were neonates and 1 was an infant. The most common source of isolation is urine and blood.A total of 35/37 isolates showed moderate to heavy growth on the SAM, while 2 isolates showed mild growth after 72 h. But all the other Candida species and other yeasts tested were inhibited on this medium. All the isolates of C. auris grew as cream to pinkish purple colonies on Hichrome agar Candida. On Tetrazolium reduction agar, all of them formed maroon colonies.The average duration of hospital stay was 25 days (range 4-65). A total of 35 of the patients were admitted to ICU, 8 had undergone mechanical ventilation and intubation. Central venous catheter was inserted in 9 patients and post-operative catheter placed in 6 patients; 4 patients had undergone tracheostomy and 25 of them had undergone some other invasive procedure. Total parenteral nutrition was received by 3 patients, 16 were diabetics and 11 were hypertensives. Prior antifungal exposure was present in 9 patients and 26 had received broad-spectrum antibiotics.The crude mortality rate with C. auris infection in patients was 32.43% and the attributable mortality rate, as considered by the treating physician was 10.81%.Antifungal resistance was noted to be amphotericin B (n = 15, 40.5%), fluconazole (n = 30, 81.1%), voriconazole (n = 4, 10.81%), itraconazole (n = 6, 16.21%), posaconazole (n = 5, 13.51%), caspofungin (n = 4, 10.81%). Multidrug resistance was noted in 15 (40.54%) isolates and 3 isolates (5.4%) were resistant to a drug from all three groups.Conclusion C. auris poses a great threat to immunocompromised individuals and those admitted in ICUs for long term.

Raman spectroscopy as a novel technique for the identification of pathogens in a clinical microbiology laboratory

Raman spectroscopy is a vibrational spectroscopic technique with which the molecular fingerprint of a microorganism can be elucidated in 1 hr or less. In our study, the utility of this method as a rapid technique for the detection of different pathogens in a clinical microbiology laboratory was tested based on their unique spectra. The reference strains from American Type Culture Collection for frequently encountered pathogens such as Staphylococcus aureus, Escherichia coli, Acinetobacter baumannii, and Pseudomonas aeruginosa were used to capture the signature spectra of these pathogenic species using confocal Raman microscopy. Furthermore, a representative reference library was constructed using the spectra of these standard strains as well as the clinical isolates. The study revealed unique spectral outputs for each bacterial species studied. Importantly, the entire procedure could be performed in an hour. With the development of an extensive database, the spectra of unknown samples can be matched with those in the library, and thus, a new identification system can be developed.

Chemical profile of Nepeta cataria L. var. citriodora (Becker) essential oil and in vitro evaluation of biological activities

Essential oil (EO) obtained by hydrodistillation in a Clevenger-type apparatus from aerial parts of Nepeta cataria L. var. citriodora (Becker), cultivated in Serbia was subjected to gas chromatography-mass spectroscopy (GC-MS) to determine the composition. Furthermore, N. cataria var. citriodora essential oil wastested to determine its antimicrobial, antioxidant, antihyperglycemic and anti-inflammatory activities in vitro.The antimicrobial activity was tested by broth microdilution method against 16 bacterial strains from American Type Culture Collection (ATCC). Four common tests for measuring in vitro antioxidant activity were used: 2, 2-diphenyl-1-picrylhydrazyl assay (DPPH), reducing power (RP), 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and β-carotene bleaching assay (BCB). Antihyperglycemic activity was examined by using α-glucosidase inhibitory potential (AHgA), while anti-inflammatory activity (AIA) was determined by protein denaturation bioassay, using egg albumin. In total, 36 compounds were isolated and detected by GC-MS technique in N. cataria var. citriodora EO. The EO is mainly comprised of oxygenated monoterpenes (93.1%), and the main compounds were two monoterpenoid alcohols, nerol (38.5%) and geraniol (24.9%), followed by two aliphatic aldehyde, geranial (14.6%) and neral (11.0%). Antimicrobial activity of this EO shows growth inhibition of all tested bacteria strains, and exhibited good antioxidant, antihyperglycemic and anti-inflammatory activities. The EO obtained from N. cataria var. citriodora grown in Serbia shows valuable biological activity, indicating its potential for use as a supplement in everyday diet and as a natural preservative in food industry.

Fracture strength of Er,Yag laser treated PMMA denture-based polymer (DBP) colonized with C. albicans, S. aureus, S.mutans, and E.coli

AimTo assess and equate the efficacy of different disinfection protocols autoclave, chlorhexidine (CHX), PDT utilizing Rose Bengal (RB), chitosan, and Er, Yag laser and their effect on fracture strength of PMMA denture-based polymer (DBP) colonized with C. Albicans, S. aureus, S.mutans, and E.coli. Material and methodsA total of 50 (n = 10) PMMA DBP were manufactured and adulterated with the American Type Culture Collection (ATCC) of diverse microbial colonies inhabited by C. Albicans, S. aureus, S.mutans, and E.coli. The specimens were subjected to different denture disinfection approaches by randomly distributing in into five groups i.e., Er, Yag laser, RB, autoclave, CHX, and Chitosan, respectively for appraising antimicrobial effectiveness. PMMA fracture load was also assessed and statistical analysis was performed for CFU/mL (log10) of exposed C. Albicans, S. aureus, S.mutans, and E.coli by one-way ANOVA and Tukey's multiple comparison test at a significance level of p < 0.05. ResultsIntergroup comparison disclosed that denture disinfection with Er, Yag laser, autoclave, Chitosan, and CHX (control) validated comparable antimicrobial efficacy to denture against all inspected CFU/mL (log10) (p>0.05). The intragroup comparison revealed that DBP sanitization with Er, Yag laser, autoclave, Chitosan, RB, and CHX substantiated equivalent effective antimicrobial efficacy in plummeting CFU/mL (log10) of S. mutans and E. coli (p>0.05) but in consideration to S.aureus and C.albicans, all groups resulted in declining their count except 5µm RB activated by PDT(p < 0.05). No significant difference was perceived in fracture load of PMMA denture base among Er, Yag laser, RB, chitosan, and CHX (control) (p > 0.05) except autoclave decontamination procedure that indicated the least fracture strength of DBP when disinfected (p < 0.05). ConclusionEr, Yag laser, and Chitosan activated by PDT have the potential to be used as an alternative to chlorhexidine for disinfecting Polymethyl methacrylate denture base as they demonstrated the highest antimicrobial efficacy against E. coli, C. Albicans, S aureus, and S. mutans with optimal fracture load.

Evaluation of antimicrobial efficacy of different hand sanitizers available in Thimphu, Bhutan

Introduction: Hand washing and the use of alcohol based hand-rub are two effective hand-hygiene procedures. Though the compliance to hand washing was relatively low, the COVID-19 pandemic situation has drastically increased the use of hand sanitizers. Thus, many hand sanitizer brands have entered the market and this study was taken up to assess the microbial efficacy of different types of hand sanitizers available in Thimphu. Methods: Ten gel based and ten liquid hand sanitizers were collected from various retail pharmacy shops in Thimphu during first quarter of 2020. Laboratory based experiment was performed using standard organism; Gram Negative Bacteria (GNB) Escherichia coli (American Type Culture Collection 25922) and Pseudomonas aurigenosa (American Type Culture Collection 27853), Gram Positive Bacteria (GPB) Staphylococcus aureus (American Type Culture Collection 25923) and a cocktail of GNB and GPC. The efficacy of hand sanitizer was assessed with the size of the Zone of Inhibition (ZoI). Results: The hand sanitizers were found to be effective against all strains of micro organism but with differentsize of ZoI. The maximum size of Zone of Inhibition was observed by sample N and Q (ZoI diameter 33.0±0.14mm) for American Type Culture Collection 25923. The Zone of Inhibition of liquid base hand sanitizers was significantly higher than that of gel base (p&lt;0.05). Conclusions: The study presents that liquid base alcohol based hand sanitizer is more efficient than the gel form.

The use of selected herbal preparations for the disinfection of Japanese quail hatching eggs

The aim of study was to evaluate microbial and hatchability traits as well as chicks quality after hatching eggs disinfection with aqueous solutions of ginger (GR), garlic (GC), oregano (O), and cinnamon (C) extracts. The experiment was divided into 2 stages, at preliminary in vitro stage antimicrobial susceptibility of plant extracts (PEs) was tested against reference strains from the American Type Culture Collection. O and GC extracts had the best antimicrobial properties (P < 0.05). Then in in vivo stage 2,400 Japanese quails hatching eggs were divided into 6 groups, 400 eggs each. Eggs from first group were not disinfected (NC, negative control), eggs from second group were disinfected by formalin fumigation (C, positive control), in other groups 5% aqueous solutions of plant extracts of GR, GC, O, C were applied by spraying respectively. After standard incubation fertility, hatchability and periodical embryonic mortality were calculated as well as the body weight and livability of chickens during 14 d of rearing. Egg disinfection by aqueous solution of PEs led to maintain the hatchability, chick weight at hatch and post hatch body weight and early mortality of birds. Exclusion of any fungal isolates on eggshell surface was induced by GC followed by O and C groups (P < 0.05). In case of the bacteria colonies reduction only GR extract was effective. Chosen plant extracts may be treated as safe and alternative substances to traditional disinfectants of hatching eggs.

PHYTOCHEMICAL AND MICROBIOLOGICAL ASPECTS OF THE &lt;i&gt;NIGELLA SATIVA&lt;/i&gt; L. HERBS STUDY

Nigella sativa L. is a promising plant object, herbal medicinal raw materials of which can be comprehensively used for the development of drugs with an antimicrobial activity.The aim of the study was to screen and compare the antimicrobial activity of water-ethanolic extractions from the Nigella sativa L. herbs with a eucalyptus tincture of as a reference preparation.Materials and methods. Chromatograms of the extracts were obtained by thin layer chromatography in the system of chloroform – ethanol – water (26:16:3). The detection of adsorption zones was carried out in daylight, in the UV light at λ=254 nm and λ=365 nm, as well as by treatment with reagents – a 3% alcohol solution of aluminum chloride and a solution of diazobenzosulfonic acid in a 20% sodium carbonate solution. The next step was to determine the minimum inhibitory concentration by the method of double serial dilutions in Mueller-Hinton nutrient broth (Bio-Rad, USA). As test cultures, the strains of the American Type Culture Collection (ATCC) microorganisms were used: Staphylococcus aureus (ATCC 29213), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), as well as Candida albicans (a clinical strain). Simultaneously, an experiment to establish a “negative” control was carried out. For the comparative evaluation of the studied samples activity, its activity was compared with the reference preparation with a proven antimicrobial activity – a eucalyptus tincture.Results. For all water-ethanolic extractions and the Nigella sativa L. herb tincture, the adsorption zones characteristic of flavonoids with Rf1 = 0.28, Rf2 = 0.15, Rf3 = 0.11 were revealed, and under the action of an alcoholic solution of aluminum chloride, the fluorescence of the adsorption zones was also enhanced, which indicates the phenolic nature of these compounds.In the course of the study, it was found out that all water-ethanolic extractions from the Nigella sativa L. herbs have the greatest antimicrobial effect against the Pseudomonas aeruginosa strain. When compared with the reference preparation – a eucalyptus tincture, it was notified that the specified tincture of the Nigella sativa L. herbs has an advantage in the antimicrobial activity over the strain of Pseudomonas aeruginosa – the action at the 16-fold dilution vs the 4-fold dilution. The action on the Escherichia coli and Candida albicans strains is comparable for the both tinctures.Conclusion. The obtained results of phytochemical and microbiological analyses will be used as a rationale for the introduction of antimicrobial preparations based on the Nigella sativa herbs in medical and pharmaceutical practice.