Evaluation of moisturizing property and antimicrobial activity of alcohol-based hand sanitizer formulations using coconut oil as a moisturizing agent against Staphylococcus aureus and Escherichia coli

During COVID-19 outbreak, alcohol-based hand sanitizer (ABHS) has been widely used for hand cleaning and removing pathogens including human pathogens and zoonosis pathogens. High concentration of alcohol induces dehydrated skin in the users. Therefore, the objective of this study was to investigate moisturizing property and antimicrobial activity of alcohol-based hand sanitizer formulations using coconut oil as a moisturizing agent against pathogens including Escherichia coli and Staphylococcus aureus. The properties including antimicrobial activity, stability of the ABHS, and satisfaction levels of the coconut oil with two existing formulations by WHO and the Ministry of Public Health of Thailand (MOPH) were determined. The formulation containing the coconut oil demonstrated antibacterial activity against both E. coli and S. aureus with the minimal inhibitory concentration (MIC) of 8.75% V/V as same as MOPH. However, the WHO-modified formulation has the highest antimicrobial activity with the MIC value of 2.19% V/V. The stability result of 3 ABHSs showed that the preserved had the same efficacy compared to that of the fresh formulations. Hand washing with coconut formulation produced an average score of 7.19±1.71 on color, 7.06±1.56 on appearance, 5.82±2.10 on odor, 6.77±1.68 on moisture, 6.88±1.42 on overall acceptance out of 9. However, the sensory analysis of these three formulations results showed no significant difference in all parameters. The data suggest that moisturizing agents tested in our study do not affect the efficacy of ethanol. The coconut oil formulation is another good option for people searching for an effective hand sanitizer for germs protection and moisturizing

Antibacterial, antibiofilm, and anti-adhesion activities of Piper betle leaf extract against Avian pathogenic Escherichia coli

Piper betle leaves have traditionally been used to treat many diseases, including bacterial infections. The present studyaimed to investigate the antibacterial, antibiofilm, and anti-adhesion activities of P. betle extract against avian pathogenicEscherichia coli (APEC). The ethanol extract of P. betle leaves demonstrated strong antibacterial activity against clinicalisolates of APEC with MIC and MBC values ranging from 0.5 to 1.0 mg/mL as compared with 1% DMSO, a negative control.Disruption and breakdown of the bacterial cells were detected when the cells were challenged with the extract at 2 × MIC.Bacterial cells treated with the extract demonstrated longer cells without a septum, compared to the control. The extract at1/8, 1/4, and 1/2× MIC significantly inhibited the formation of the bacterial biofilm of all the tested isolates except the iso-late CH10 (P < 0.05) without inhibiting growth. At 1/2 × MIC, 55% of the biofilm inhibition was detected in APEC CH09,a strong biofilm producer. At 32 × MIC, 88% of the inhibition of viable cells embedded in the mature biofilm was detectedin APEC CH09. Reduction in the bacterial adhesion to surfaces was shown when APEC were treated with sub-MICs of theextract as observed by SEM. Hydroxychavicol was found to be the major compound presented in the leaf extract as detectedby GC–MS analysis. The information suggested potential medicinal benefits of P. betle extract to inhibit the growth, biofilm,and adhesion of avian pathogenic E. coli.publishe

Anti-Acanthamoeba activity of a semi-synthetic mangostin derivative and its ability in removal of Acanthamoeba triangularis WU19001 on contact lens

Garcinia mangostana L., also known as the mangosteen tree, is a native medicinal plant in Southeast Asia having a wide variety of pharmacologically active compounds, including xanthonoid mangostin. In this study, we examined the pharmacological activities of the selected semi-synthetic mangostin derivative, namely, amoebicidal activity, encystation inhibition, excystation activity, and removal capacity of adhesive Acanthamoeba from the surface of contact lens (CL). Among the three derivatives, C1 exhibited promising anti-Acanthamoeba activity against Acanthamoeba triangularis WU19001 trophozoites and cysts. SEM images displayed morphological changes in Acanthamoeba trophozoites, including the loss of acanthopodia, pore formation in the cell membrane, and membrane damage. In addition, the treated cyst was shrunken and adopted an irregular flat cyst shape. Under a fluorescence microscope, acridine orange and propidium iodide (AO/PI) staining revealed C1 induced condensation of cytoplasm and chromatin with the loss of cell volume in the treated trophozoites, while calcofluor white staining demonstrated the leakage of cell wall in treated cysts, leading to cell death. Interestingly, at the concentration ranges in which C1 showed the anti-Acanthamoeba effects (IC50 values ranging from 0.035–0.056 mg/mL), they were not toxic to Vero cells. C1 displayed the highest inhibitory effect on A. triangularis encystation at 1/16×MIC value (0.004 mg/mL). While C1 demonstrated the excystation activity at 1/128×MIC value with a high rate of 89.47%. Furthermore, C1 exhibited the removal capacity of adhesive Acanthamoeba from the surface of CL comparable with commercial multipurpose solutions (MPSs). Based on the results obtained, C1 may be a promising lead agent to develop a therapeutic for the treatment of Acanthamoeba infections and disinfectant solutions for CL

Targeting Acanthamoeba proteins interaction with flavonoids of Propolis extract by in vitro and in silico studies for promising therapeutic effects

Background: Propolis is a natural resinous mixture produced by bees. It provides beneficial effects on human health in the treatment/management of many diseases. The present study was performed to demonstrate the anti-Acanthamoeba activity of ethanolic extracts of Propolis samples from Iran. The interactions of the compounds and essential proteins of Acanthamoeba were also visualized through docking simulation. Methods: The minimal inhibitory concentrations (MICs) of Propolis extract against Acanthamoeba trophozoites and cysts was determined in vitro. In addition, two-fold dilutions of each of agents were tested for encystment, excystment and adhesion inhibitions. Three major compounds of Propolis extract such as chrysin, tectochrysin and pinocembrin have been selected in molecular docking approach to predict the compounds that might be responsible for encystment, excystment and adhesion inhibitions of A. castellanii. Furthermore, to confirm the docking results, molecular dynamics (MD) simulations were also carried out for the most promising two ligand-pocket complexes from docking studies. Results: The minimal inhibitory concentrations (MICs) 62.5 and 125 µg/mL of the most active Propolis extract were assessed in trophozoites stage of Acanthamoeba castellanii ATCC30010 and ATCC50739, respectively. At concentrations lower than their MICs values (1/16 MIC), Propolis extract revealed inhibition of encystation. However, at 1/2 MIC, it showed a potential inhibition of excystation and anti-adhesion. The molecular docking and dynamic simulation revealed the potential capability of Pinocembrin to form hydrogen bonds with A. castellanii Sir2 family protein (AcSir2), an encystation protein of high relevance for this process in Acanthamoeba. Conclusions: The results provided a candidate for the development of therapeutic drugs against Acanthamoeba infection. In vivo experiments and clinical trials are necessary to support this claim

ฤทธิ์การยั้บยั้งแบคทีเรีย Streptococcus pneumoniae ของโรโดไมรโทน โดยการวิเคราะห์ทางโปรติโอมิกส์และเมตาโบโลมิกส์

Thesis (Ph.D., Microbiology)--Prince of Songkla University, 201

First-lactation milk fat-to-protein ratio in tropically-raised dairy cows: environmental and genetic influences

The objective of this research was to determine factors influencing to fat-to-protein ratio (FPR) in the first-lactation of dairy cows raised in tropical climate. The dataset included fat percentage, protein percentage, and FPR values from 160,506 records representing 23,201 first-lactation cows that calved between 1993 and 2017. These cows originated from 508 farms in all regions of Thailand. The data were analyzed using the general linear model procedure in the R program. The models included herd-year-season of records, and breed group as fixed effects, and age at calving and days in milk as covariates. The result showed that the average fat percentage, protein percentage, and FPR were 3.560±0.960, 3.130±0.390 and 1.150±0.330, respectively. Effects of herd-year-season of records, breed group, age at calving, and days in milk were highly significant for all traits (P<0.05). Cows containing less than 87.50% Holstein Friesian (HF) blood gave fat percentage, protein percentage and FPR higher than 93.75%HF and 87.50-93.74%HF. The FPR was highest in the winter and lowest in the summer. In conclusion, present results indicated that fat percentage, protein percentage and FPR are clearly influenced by both genetic and non-genetic factors. Therefore, development of different breed, feeding systems, according to season and herd is needed to produce quality of fat and protein percentage

Molecular characterization and nucleotide substitution of antibiotic resistance genes in multidrug-resistant Escherichia coli isolated from environmental swine farms

An increase of multi-drug resistant Escherichia coli in swine husbandry have been concerned worldwide. This study aimed to determine the multidrug resistance and nucleotide substitution of beta-lactam antibiotic and tetracycline resistant genes in E. coli from swine farms in Southern Thailand. A total of 112 isolates of E. coli was isolated from 50 pig farms, which were confirmed by identified by MALDI-TOF analysis. Seventy-three isolates (65.18%) and 39 isolates (34.82%) were isolated from the feces and waste water samples, respectively. One hundred percent resistance to beta-lactam antibiotics as well as their resistant gene blaTEM was detected in isolates. Furthermore, 81% isolates were tetracycline resistance and both tetA [68.42% (13/19) in feces samples, 72.73% (8/11) in waste water samples] and tetB [10.53% (2/19) in feces samples, 18.18% (2/11) in waste water samples] genes responsible for tetracycline resistance were observed. Furthermore, 54 isolates had multi-drug resistance that presented 11 different patterns. The nucleotide substitution of genes was detected in 3 isolates of E. coli, and may consider as the point mutation. The nucleotide at 859 bp of tetA gene of the isolate WU-WW009-01 was changed from T to A. While, the isolate WU-WW004-02 showed 2 nucleotide substitution sites at the position of 266 (from A to G) and 859 (from T to G) bp. The nucleotide at 36 bp of blaTEM gene of the isolate WU-F003-02 was replaced from G to A. Findings of this study may help to control the spread of E. coli antibiotic resistance genes in the swine farms

Antifungal activity of protein hydrolysates from Thai Phatthalung Sangyod rice (Oryza sativa L.) seeds

Background and Aim: Fungal zoonoses are an economic and public health concern because they can cause various degrees of morbidity and mortality in animals and humans. To combat this issue, alternative natural antifungals, such as products derived from rice protein hydrolysates or rice antifungal protein/peptide are being considered because they are highly bioactive and exhibit various functional properties. Thailand is a leading rice producer and exporter. Among the various cultivated rice varieties, Sangyod rice (Oryza sativa L.) is exclusively indigenous to Thailand’s Phatthalung province; it has a Thai geographical indication tag. Here, we investigated whether the Phatthalung Sangyod rice seeds have bioactive antifungal peptides. Materials and Methods: Antifungal activity in four Sangyod rice seed extracts (SYPs) – namely, (1) the crude lysate, SYP1; (2) the heat-treated lysate, SYP2; (3) the heat- and pepsin digested lysate, SYP3; and (4) the heat- and proteinase K-digested lysate, SYP4 – was analyzed. Protein concentrations in these SYPs were determined using the Bradford assay. The total phenolic compound content was determined using the modified Folin–Ciocalteu method in a 96-well microplate. Then, the SYP protein pattern was determined using the sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Subsequently, using the agar well diffusion method, the antifungal properties of these SYPs were tested against ten medically important pathogenic fungi. The minimal inhibitory concentration (MIC) and minimal fungicidal concentration values were determined for the active SYPs – SYP2-4. Finally, the clinical safety of SYP4 was determined using a hemolytic assay (using canine red blood cells [RBCs]). Results: The crude lysate SYP1 did not show antifungal activity against any of the ten tested pathogenic fungi. Surprisingly, hydrolysates SYP2, SYP3, and SYP4 displayed antifungal properties against the ten tested pathogenic fungi. Thus, heat and enzymatic hydrolysis seem to transform the bioactivity of the crude protein extract – SYP1. Further, SYP4 shows the most effective antifungal activity. It completely inhibited Cryptococcus neoformans, Talaromyces marneffei yeast phase, Trichophyton mentagrophytes, and Trichophyton rubrum. A partial inhibitory action on Candida albicans and Microsporum gypseum was possessed while showing the least activity to C. neoformans. SYP4 was nontoxic to canine RBCs. Hemolysis of canine RBCs was undetectable at 1 × MIC and 2 × MIC concentrations; therefore, it can be safely used in further applications. Conclusion: These results indicate that heat and proteinase K hydrolyzed SYP is a very potent antifungal preparation against animal and human fungal pathogens and it can be used in future pharmaceuticals and functional foods

Lyophilized cell-free supernatants of Lactobacillus isolates exhibited antibiofilm, antioxidant, and reduces nitric oxide activity in lipopolysaccharide-stimulated RAW 264.7 cells

Background Probiotics can release bioactive substances known as postbiotics, which can inhibit pathogenic microorganisms, improve immunomodulation, reduce antioxidant production, and modulate the gut microbiota. Methods In this study, we evaluated the in vitro antimicrobial effects, antioxidant activity, and anti-inflammatory potential of 10 lyophilized cell-free supernatants (LCFS) of Lactobacillus isolates. LCFS was obtained via centrifugation and subsequent lyophilization of the supernatant collected from the culture medium ofeach isolate. The antibacterial and antibiofilm activities of the LCFS were determined using broth microdilution. The antioxidant potential was evaluated by measuring the total phenolic and flavonoid contents and 2,2-Diphennyl-1-picrylhydrazyl (DPPH) and 2,2’-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS+) radical scavenging activities. Results All the isolates were able to inhibit the four tested pathogens. The isolates exhibited strong antibiofilm activity and eradicated the biofilms formed by Acinetobacter buamannii and Escherichia coli. All the prepared Lactobacillus LCFS contained phenols and flavonoids and exhibited antioxidant activities in the DPPH and ABTS+ radical scavenging assays. The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay revealed that LCFS was not cytotoxic to RAW 264.7 cells. In addition, the ten Lactobacillus LCFS decreased the production of nitric oxide. Conclusions All the isolates have beneficial properties. This research sheds light on the role of postbiotics in functional fermented foods and pharmaceutical products. Further research to elucidate the precise molecular mechanisms of action of probiotics is warranted