Evaluation of a Rhodomyrtus tomentosa ethanolic extract for its therapeutic potential on Staphylococcus aureus infections using in vitro and in vivo models of mastitis.

An ethanolic extract from Rhodomyrtus tomentosa leaves (RTL) was studied as a natural alternative to control Staphylococcus aureus, which is an important pathogen responsible for bovine mastitis. The minimal inhibitory concentrations (MICs) of the RTL extract and of rhodomyrtone, a pure compound isolated from the plant, were determined by a microdilution method. Rhodomyrtone and the RTL extract exhibited antibacterial activity against S. aureus, including its persistent phenotype (SCV: small-colony variant) and a biofilm hyperproducer strain, with MICs of 0.25-0.5 and 8-16 µg/mL, respectively. Time-kill kinetics showed a strong bactericidal activity for both the RTL extract- and rhodomyrtone-treated bacteria at 2 × MIC as early as 4 h post-exposure. An additive effect of the extract at 0.5 × MIC was observed in a combination with oxytetracycline or pirlimycin against S. aureus by showing a 64- to 128-fold reduction in antibiotic MICs. Moreover, the RTL extract significantly decreased the number of intracellular SCVs inside bovine mammary epithelial cells. However, the extract or its combination with pirlimycin only slightly improved the activity of pirlimycin against the bacterial colonization of mouse mammary glands. In vitro MICs determined in the presence of casein indicated that the limited activity of the RTL extract in the murine model of mastitis could be linked to neutralization of active components by milk proteins. While the RTL extract showed interesting antibacterial properties in vitro, to be considered as an alternative to antibiotics in dairy farms, formulation studies are needed to cope with the observed reduction of activity in vivo.

Detection of Microplastics in Human Breast Milk and Its Association with Changes in Human Milk Bacterial Microbiota.

Background: Presently, there is increasing public consciousness regarding the contamination and detection of microplastics (MPs) within the human body, and studies on the detection and characterization of MPs in human breast milk are limited. Objectives: This study aims to investigate the prevalence and characteristics of MPs found in human breast milk and examine the relationship between maternal hygiene practices, complications that may arise during breastfeeding, and the composition of the bacterial microbiota. Methods: Postpartum breast milk was analyzed for MPs using Raman micro-spectroscopy. The relationship between MP detection, maternal hygiene, breastfeeding complications, and bacterial microbiota was examined. In order to identify correlations and differences between groups that had detected and non-detected MPs, statistical analyses were performed, which involved demographic comparisons and correlation network analysis. Results: The mean age of the 59 postpartum women was 28.13 years. We found MPs in 38.98% of breast milk samples (23 of 59), exhibiting diverse morphological and chemical characteristics. Most MP polymers were polypropylene, polyethylene, polystyrene, and polyvinyl chloride. Maternal hygiene and breastfeeding complications differed between the MPs-detected and non-detected groups. Maternal behaviors may influence the presence of microplastics in breast milk, which were associated with these differences. Bacterial microbiota analysis revealed significant taxonomic differences between the MPs-detected and non-detected groups. Staphylococcus and Streptococcus dominated the MPs-detected group, while Enterobacter, Escherichia, Pseudomonas, and Acinetobacter dominated the non-detected group. The MPs-detected group had a more even bacterial distribution, especially Bacteroides. Conclusions: This study found MPs in 38.98% of breast milk samples using Raman micro-spectrometry, with PP, PE, and PVC being the most common. Significant differences in maternal hygiene and breastfeeding complications were found between the groups with and without MPs. Breast milk microbiota may be linked to MP detection. Further study should be conducted to identify the possible maternal-child health.

Potential Activities of Centella asiatica Leaf Extract against Pathogenic Bacteria-Associated Biofilms and Its Anti-Inflammatory Effects.

The medicinal value of Centella asiatica leaf extract was evaluated as an alternative treatment. The chemical composition of the leaf extract was analyzed, and the biological activities were determined. High-performance liquid chromatography coupled with a photodiode array detector (HPLC-PDA) was used to identify the asiatic acid, madasiatic acid, and madecassic acid/Brahmic acid isolated from the ethanolic extract. The plant extract at 25 mg/disk was found to inhibit both Gram-positive and Gram-negative pathogenic bacteria by the agar disk diffusion test. The MIC and MBC of the ethanolic extracts were better than those of the aqueous extracts. The ethanolic extracts showed antibacterial activity against Gram-positive bacteria with MICs and MBCs ranging from 1.024 to 2.048 mg/mL and 2.048 to 4.096 mg/mL, respectively. The remarkable antibacterial activities were observed against S. mutans. The ethanolic extract at a concentration of 1/2 × MIC exhibited the inhibition effect on S. mutans biofilm formation like the activity of 0.2% chlorhexidine and significantly modified hydrophobicity of the bacterial cell surface. The effects were confirmed via molecular docking analysis. The binding affinities of asiatic acid, madecassic acid, and madasiatic acid with glucosyltransferase C (GtfC) of S. mutans exhibited superior strength in comparison with alpha-acarbose and chlorhexidine. Moreover, the nitric oxide (NO) secretion of RAW247.6 cells was determined after treating the cells with concentrations of the extract. The C. asiatica ethanolic extract can inhibit the secretion of NO, which can inhibit the inflammatory process. The findings indicate the applications of the C. asiatica ethanolic extract as the alternative anti-S. mutans agent and could be used for further formulation for the treatment and prevention of dental diseases and inflammatory injury in the oral cavity.

Efficacy of topical local anesthetic, topical cooling spray, and audiovisual distraction on relief of needle-related pain during blood collection: A randomized controlled trial.

BACKGROUND: Venipuncture is one of the most common invasive procedures in healthcare, often resulting in the experience of pain. While audiovisual distraction, topical anesthesia and cold spray application have been reported as methods to reduce pain, there is a lack of studies that focus on comparing their efficacy and safety. OBJECTIVES: We aimed to compare the efficacy and safety of pain reduction during venipuncture using audiovisual distraction, topical anesthesia and cold spray application. MATERIAL AND METHODS: A randomized controlled study was conducted at Walailak University (Nakhon Si Thammarat, Thailand) from April 2023 to July 2023. Eligible adult participants voluntarily enrolled in the study and were randomly assigned to 1 of 4 groups: group 1 (control), group 2 (topical anesthetic), group 3 (cooling spray), and group 4 (audiovisual distraction). Pain scores and satisfaction levels were assessed following the venipuncture procedure on the upper extremities. RESULTS: Forty-seven participants were included in the final analysis. The participants had a mean age of 42.3 years (standard deviation (±SD): 13.1), with the majority being female (66.0%). The participants in the intervention groups reported lower pain scores than those in group 1. The mean differences were 2.67 points in group 2 (95% confidence interval (95% CI): 1.49-3.84; p < 0.001), 1.56 points in group 3 (95% CI: 0.15-2.98; p = 0.077), and 1.67 points in group 4 (95% CI: 0.37-2.96; p = 0.042). However, the pain reduction did not reach statistical significance when comparing these 3 interventions. All groups reported a median satisfaction level of 3, with no significant difference among them (H(3) = 6.050, p = 0.109). CONCLUSIONS: Pain reduction interventions, including topical anesthetic, cooling spray and audiovisual distraction, are effective methods for alleviating pain during venipuncture. Participants who received a topical anesthetic reported the lowest pain scores and highest levels of satisfaction.

Chicago sky blue gel for better visualization of Demodex in patients with Demodex blepharitis.

Background: Demodex blepharitis is a common chronic disease. The number of mites is associated with ocular discomfort. The accurate number derived from well-stained specimens is, hence, in favor of diagnosing, monitoring, and determining treatment responses. Methods: A cross-sectional study was conducted between April and July 2022 at the dermatology and ophthalmology clinic, Walailak University, Thailand. Adult participants with clinical suspicion of Demodex blepharitis were recruited. We examined eyelashes under light microscopy to quantify the number of Demodex mites before and after adding CSB gel. The mite counts, evaluated by an untrained investigator and an experienced investigator, were recorded and compared. Results: A total of 30 participants were included for final analysis, among which 25 (83.3%) were female. The median age was 64.0 years (IQR, 61.0-68.0). The median Demodex counts evaluated by the experienced investigator before and after adding CSB gel were 1.0 (IQR, 0.0-1.0) and 2.5 (IQR, 2.0-3.0), respectively (p < 0.001). Moreover, the median Demodex counts evaluated by the untrained investigator before and after adding CSB gel were 1.0 (IQR, 0.0-1.0) and 2.0 (IQR, 1.0-3.0), respectively (p < 0.001). The correlation coefficient between Demodex counts after the addition of CSB counted by the experienced investigator and those counted by the untrained investigator was 0.92 (p < 0.001). CSB gel is a promising product to identify and quantify the number of Demodex mites. The findings supported the consideration of CSB gel as one of the diagnostic stains.

The potency of herbal extracts and its green synthesized nanoparticle formulation as antibacterial agents against Streptococcus mutans associated biofilms.

This study aims to determine the effects of the extracts of Streblus asper, Cymbopogon citratus, Syzygium aromaticum and its formulation of green synthesized silver nanoparticle (AgNPs) on Streptococcus mutans growth and biofilm formation. The ethanolic extracts of S. asper, C. citratus, S. aromaticum, and a mix of the three herbs demonstrated antibacterial activity against S. mutans isolates by reducing bacterial biofilm formation and decreasing bacterial cell surface hydrophobicity. The formulated AgNPs from the ethanolic extracts could enhance the antibacterial activities of the plant extracts. Molecular docking found the best interaction between luteolin isolated from C. citratus and glucosyltransferase protein (GtfB), assuming the promising anti-biofilm activity. The scanning electron microscopy revealed morphological changes in the biofilm structure and a significant decrease in the biofilm area of the AgNPs treated. The study suggested that the extracts and its application could be used as natural alternative agents with multi-action against S. mutans infections.

Effectiveness of an online educational video intervention to improve the knowledge and behavior of contact lens care during the COVID-19 pandemic: A pre-test/post-test design.

Objective: To assess the effectiveness of an online educational video in improving contact lens (CL) care knowledge and behavior. Methods: Participants completed a 47-item questionnaire on their CL hygiene knowledge and wear and care behavior. A 5-min CL educational video was shown, and participants completed a post-test. After 2 months, the same questionnaire was used to determine knowledge retention and behavioral changes. Descriptive statistics and McNemar's tests were performed. Results: The mean age of the 132 enrolled participants was 24 years, and 61% were female. The knowledge scores significantly improved after watching the educational video (p < 0.001). Two months after the intervention, the participants reported changes in their hygienic behavior (p < 0.001). Conclusions: Online video-based learning is an effective educational tool for improving the knowledge and behavior of CL care. Practice implications: Patient education via online videos is an innovative and successful strategy that raises awareness, increases patient knowledge, and encourages preventative health behavior to avoid CL-related complications.

Bacterial microbiota of the contact lens surface and associated care behaviours.

Introduction: Contact lens (CL) wear has been reported to cause changes to the microbiome of the ocular surface. More insight into the alteration of this microenvironment can help to understand the pathogenesis of CL-related eye infections. Knowledge of the relationship between the CL wearer's behaviours and pathogens would help health care providers focus on each step of proper CL care. This study aims to determine the behaviours that might be associated with the community of bacteria on CL. Methods: A cross-sectional design was performed using anonymous questionnaires to obtain demographic data and assess hygiene practices among volunteering wearers. The CLs used were collected to evaluate the prevalence of pathogenic bacteria associated with ocular infections by PCR and microbiota analysis. Results: The bacterial microbiota study revealed a total of 19 genera and 26 isolated strains from 20 eligible CLs. Enterobacter, Staphylococcus, and Achromobacter were the main genus in this subject population. Staphylococcus pasteuri and Achromobacter agilis were the most common pathogens at 65% and 35%, respectively. Enterobacter mori, a nonpathogenic organism, was found to be the most predominant strain, accounting for 27.51% of the total bacterial constituents. The risk behaviour of CL wear that was significantly associated with A. agilis contamination was cleaning the CL case with tap water (P value = 0.04). Conclusions: This is the first study focusing on the association between the culture selected microbial community on the CL surface and compehensive behavioural characteristics. Environmental contamination was the main source of microbes found on CL surfaces. An emphasis in patient education should be placed on careful handling during the CL care routine and managing the hygiene of the surroundings.

Rub and Rinse Contact Lenses Before Wearing as a Protective Regimen Against Contact Lens-Related Eye Infections.

Purpose: Data on contact lens (CL) users in Thailand are limited, and previous reports have mainly focused on young populations. This study aims to determine demographic data, hygienic behaviour, and complications related to the usage of CLs in the general population. Methods: This was a cross-sectional survey conducted from November 2020 to March 2021 using an anonymous questionnaire. Information on the demographic traits of the participants, characteristics of CL use, and CL care behaviours was collected. The responses were summarised and presented as a total behaviour score. Regression analysis was performed to evaluate the potential risk factors for a history of CL-related eye infections. Results: A total of 134 CL wearers were recruited. Of all the participants, 83.58% were female and 16.42% were male. The average age was 31.14±10.69 years old. The total behaviour score showed that 62.69% of the population had good CL behaviour, while 37.31% exhibited poor CL behaviour. More than 50% of participants practised appropriate CL handling. Common unfavourable behaviours (>50%) reported were not using CL care solution to clean the storage case, not rubbing and rinsing the lens before storage, exposure to water during wear, use of makeup around the ocular area, and missing annual eye exams. Other less common but important instances of mishandling included using tap water to clean the lenses, topping off old CL solution, and exceeding the recommended planned-replacement period. However, no behaviours were significantly associated with a history of CL-related eye infections. Rubbing and rinsing CLs before wearing them was found to be a protective factor against a history of eye infections. Conclusion: Rubbing and rinsing CLs before wearing should be mentioned as an additional recommendation for the user.

Influence of Contact Lens Materials and Cleaning Procedures on Bacterial Adhesion and Biofilm Formation.

PURPOSE: The present study aimed to compare the existing soft contact lens (CL) materials regarding their influence on bacterial biofilm formation and adhesion susceptibility. Then, the study was designed to investigate the effectiveness of various disinfecting solutions and evaluate the ability of cleaning regimens in terms of anti-bacterial adhesion and biofilm removal on different soft CL materials. METHODS: Bacterial biofilm formation on CLs was evaluated by biomass assay. Adhesion assay and standard plate count were carried out at time-interval periods within 24 h. Various CL disinfecting procedures were assessed for their efficacy to remove biofilm and reduce bacterial adhesion. Scanning electron microscopy (SEM) was performed for the morphological assessment of bacterial biofilm. RESULTS: Printed hydroxyethyl methacrylate (HEMA) CLs significantly demonstrated more biofilm staining and bacterial attachment when compared with non-printed HEMA CLs, while the Filcon II 3 and Nesofilcon A CLs possessed less biofilm biomass and adherent cells. Staphylococcus aureus and Pseudomonas aeruginosa represented the highest biofilm producing bacteria on HEMA-based materials in this study. The disinfecting regimen with the highest efficacy was the two-step system, first using multipurpose disinfecting solution containing edetate disodium and sorbic acid (MPDS+EDSA), followed by soaking in multipurpose solution (MPS). The regimen demonstrated the greatest effect against the pre-formed biofilm and the adhesion activity of S. aureus and P. aeruginosa on the soft CLs. The SEM micrographs confirmed the morphological changes of bacterial biofilm after disinfecting and revealed that the two-step system treated CLs displayed less adherent bacteria. CONCLUSION: HEMA-based soft CLs may facilitate bacterial biofilm formation and adhesion capability. The two-step system was the most effective regimen for biofilm removal, where the soaking period in the disinfecting solution of the no-rub regimen should last more than 6 h to remove pre-formed biofilm.

Rhodomyrtus tomentosa (Aiton) Hassk. leaf extract: An alternative approach for the treatment of staphylococcal bovine mastitis.

Antibiotic residues in dairy products as well as emergence of antimicrobial resistance in foodborne pathogens have been recognized as global public health concerns. The present work was aimed to study a potent antibacterial extract from natural product as an alternative treatment for staphylococcal bovine mastitis. Staphylococcal isolates (n=44) were isolated from milk samples freshly squeezed from individual cows. All staphylococcal isolates were resistant to ampicillin, ciprofloxacin, erythromycin, gentamicin, penicillin, except vancomycin. Rhodomyrtus tomentosa leaf ethanolic extract was accessed for its antibacterial activity and anti-inflammatory potential. The extract exhibited profound antibacterial activity against all of staphylococcal isolates with MIC and MBC values ranged from 16-64 µg/ml and 64->128 µg/ml, respectively. Moreover, the extract also exerted anti-protein denaturation and human red blood cell membrane stabilizing activity. The results support the use of R. tomentosa extract that could be applied to cure bovine mastitis and to reduce inflammatory injury caused by the bacterial infections.

Effects of Rhodomyrtus tomentosa Leaf Extract on Staphylococcal Adhesion and Invasion in Bovine Udder Epidermal Tissue Model.

Bovine mastitis is one of the most important infectious diseases in dairy herds, and staphylococci are the most important etiologic agents of this disease. Antibiotics and chemical agents used in livestock for prevention and cure of the disease can accumulate in milk and give rise to food safety concerns. Rhodomyrtus tomentosa leaf extract was studied as an alternative approach to reduce the bacterial infections. The ethanolic extract of this plant demonstrated antibacterial activity with minimum inhibitory concentration (MIC) values as low as 16-64 µg/mL against staphylococcal isolates. In addition, the extract had an effect on the bacterial cell surface properties by increasing its hydrophobicity in a concentration dependent manner. To further extend the antibacterial efficacy, silver nanoparticles synthesized with the extract, a pure rhodomyrtone, and liposomal encapsulated rhodomyrtone were applied and their inhibitory effects on bacterial adhesion and invasion were determined by ex vivo study in a bovine udder epidermal tissue model. These agents exerted remarkable antibacterial activity against staphylococci and decreased the adhesion of the bacterial cells to the tissues. These results supported that R. tomentosa ethanolic extract could be applied as an alternative agent for bovine udder care in dairy farms.