Methicillin-resistant Staphylococcus Aureus Bacteria Research Articles

Antibacterial Activity by Diffusion and Dilution Methods of Liquid Smoke Cocos nucifera L.

Introduction: Liquid smoke is a product that obtained industrially from coconut shell through pyrolysis process at 400°C for 2,5 hours and redestillation to obtain grade 1 liquid smoke. Coconut shell liquid smoke contains chemical compounds, namely phenol, which has been researched to act as a disinfectant that has broad spectrum antibacterial activity against gram positive and gram negative bacteria. Method: This study aims to determine the potential of cocos nucifera L. liquid smoke as an antibacterial against Staphylococcus aureus and Methicillin-resistant staphylococcus aureus (MRSA) bacteria using diffusion and dilution methods. The concentrations used were 100%, 80%, 60%, 40%, 35%, 30%, 15%, 10%, 7%, 5%. As positive control was used Tetracycline 25 ppm. Results: Liquid smoke has the potential to inhibit the growth of both bacteria. The effective concentration to inhibit the growth of S. aureus at 40% concentration and MRSA at 80% concentration. The MIC value of liquid smoke against S. aureus is 0,78%, while on MRSA 3,12%. The MBC of S. aureus and MRSA bacteria at concentrations 1,56% and 6,25%. Staphylococcus aureus has higher sensitivity to Cocos nucifera L. grade 1 liquid smoke than MRSA bacteria. Conclusion: Cocos nucifera L. liquid smoke has antibacterial activity against Staphylococcus aureus and MRSA at effective concentrations of 40% and 80%. The MIC value of S. aureus is 0,78%, while MRSA is 3,12%. The MBC of S. aureus and MRSA bacteria at concentrations 1,56% and 6,25%. Keywords: antibacterial, liquid smoke, phenol, Cocos nucifera L.

Antibacterial activity of Kemloko fruit ethanol extract (Phyllanthus emblica L.) against Staphylococcus aureus and MRSA bacteria

Background: Indonesia is a tropical country, so the prevalence of infectious diseases caused by bacteria remains high. On the other hand, the high use of antibacterial ingredients in Indonesia causes bacterial resistance to existing antibacterial drugs. One of the herbal plants that some people widely use is Kemloko fruit. This study aimed to determine the activity of Kemloko fruit (Phyllanthus emblica L.) ethanol extract against the growth of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA) bacteria. Methods: Simplicia of Kemloko fruit were tested for secondary metabolites and characterization test of the simplicia. The antibacterial activity test was carried out using the diffusion method, where the disc paper was immersed in the test solution with the ratio of the test concentration. Results: The results of phytochemical screening showed that the secondary metabolite compounds of Kemloko fruit are alkaloids, flavonoids, steroids, tannins, saponins, and glycosides. Antibacterial activity of ethanol extract of Kemloko fruit with agar diffusion method effectively inhibits S. aureus bacteria, namely at a concentration of 80 mg/ml, the inhibition zone is 14.1 mm, and at a concentration of 100 mg/ml, MRSA bacteria obtained an inhibition zone of 14.3 mm. Conclusions: The ethanol extract inhibits the growth of S. aureus and MRSA.

Turning Polysaccharides into Injectable and Rapid Self-Healing Antibacterial Hydrogels for Antibacterial Treatment and Bacterial-Infected Wound Healing.

Great efforts have been devoted to the development of novel and multifunctional wound dressing materials to meet the different needs of wound healing. Herein, we covalently grafted quaternary ammonium groups (QAGs) containing 12-carbon straight-chain alkanes to the dextran polymer skeleton. We then oxidized the resulting product into oxidized quaternized dextran (OQD). The obtained OQD polymer is rich in antibacterial QAGs and aldehyde groups. It can react with glycol chitosan (GC) via the Schiff-base reaction to form a multifunctional GC@OQD hydrogel with good self-healing behavior, hemostasis, injectability, inherent superior antibacterial activity, biocompatibility, and excellent promotion of healing of methicillin-resistant Staphylococcus aureus (MRSA)-infected wounds. The biosafe and nontoxic GC@OQD hydrogel with a three-dimensional porous network structure possesses an excellent swelling rate and water retention capacity. It can be used for hemostasis and treating irregular wounds. The designed GC@OQD hydrogel with inherent antibacterial activity possesses good antibacterial efficacy on both S. aureus (Gram-positive bacteria) and Escherichia coli (Gram-negative bacteria), as well as MRSA bacteria, with antibacterial activity greater than 99%. It can be used for the treatment of wounds infected by MRSA and significantly promotes the healing of wounds. Thus, the multifunctional antibacterial GC@OQD hydrogel has the potential to be applied in clinical practice as a wound dressing.

Combination of curcuminoid and collagen marine peptides for healing diabetic wounds infected by methicillin-resistant Staphylococcus aureus

Background and Aim: The high prevalence of diabetes mellitus in Indonesia indirectly reflects the high risk of developing chronic wounds that are susceptible to infection. Methicillin-resistant Staphylococcus aureus (MRSA) is an infectious pathogen that is resistant to various antibiotics. Therefore, antibiotic therapy is ineffective enough to treat chronic hyperglycemic wounds caused by MRSA infection. Curcuminoids have anti-inflammatory and antibacterial effects by inhibiting the enzymatic pathways involved in the pathogenesis of inflammation. Collagen is a tissue regeneration inducer. The combination of these two ingredients is expected to be an alternative therapy for MRSA-infected hyperglycemic chronic wounds without the risk of antibiotic resistance. The aim of this study was to investigate the efficacy of hydrolacin-gel in wound healing and inhibiting the growth of MRSA bacteria, as well as to determine the optimal concentration of curcuminoids combined with collagen marine peptides (CMPs). Materials and Methods: Hydrolacin-gels were prepared by homogenizing curcuminoid nanoemulsions and CMPs. The evaluation of preparation includes stability tests and antibacterial activity tests. Wound diabetic mice were treated with various combinations of curcuminoid and CMPs. Wound healing was observed based on malondialdehyde levels as a marker of oxidative stress and histopathological changes in the skin wound. Results: Hydrolacin-gel was formulated by combining curcuminoid nanoemulsion (more water soluble) and CMPs, with the ratio of formula 1 (1:2, curcuminoid 43.3 mg and CMPs 5.58 mg), formula 2 (1:1, curcuminoid 86.8 mg and CMPs 3.72), and formula 3 (2:1, curcuminoid 130.2 mg and CMPs 1.87 mg) calculated based on the effective dose of curcuminoid 200 mg/kg body weight (BW) and CAMPs 0.9 g/kg BW. Hydrolacin-gel had a potential antibacterial activity against MRSA. Hydrolacin-gel induced wound tissue repair and reduced oxidative stress caused by inflammation in diabetic-infected MRSA. Hydrolacin-gel could be used for healing MRSA-infected diabetic wounds, especially formula 3 with the ratio of curcuminoid: CMPs = 2:1. Conclusion: Hydrolacin-gel combining curcuminoid nanoemulsion and CMPs effectively inhibited the inflammatory process and increased re-epithelialization in MRSA-infected diabetic wound healing. Hydrolacin-gel with curcuminoid (130.2 mg) and CMPs (1.87 mg) at a concentration ratio of 2:1 appeared to be the best formula against MRSA infection in diabetic wounds. Keywords: collagen marine peptides, curcuminoids, diabetic, methicillin-resistant Staphylococcus aureus, wound.

Effectiveness of Lemongrass Leaf Ethanol Extract Spray Preparation in Inhibiting the Growth of Diabetic Ulcer Isolate MRSA Bacteria

Lemongrass (Cymbopogon nardus) is a type of grass from the order Graminales which is typical of tropical areas of Asia. Compounds contained in lemongrass leaves can inhibit the growth of Methicillin Resistant Staphylococcus aureus (MRSA) bacteria. This research aims to determine the effectiveness of spray preparations of lemongrass leaf extract in inhibiting the growth of MRSA bacterial isolates from diabetic ulcers. The research design of this study is quasi experimental. Data obtained from primary data by testing the inhibitory power of spray preparations against MRSA bacteria. The results obtained were analyzed using the One Way Anova statistical test. The diameter of the inhibition zone for MRSA bacteria in the 25% concentration ethanol extract spray preparation was 4 mm, the 35% concentration was 7 mm, the 45% concentration was 11 mm and the antibiotic Vancomycin was 12 mm. There are differences in the inhibition zone of spray preparations of ethanol extract of lemongrass leaves with concentrations of 25%, 35%, 45% and the antibiotic disc vancomycin with p value = 0.00 < α 0.05. Spray preparations of ethanol extract of lemongrass leaves with concentrations of 25%, 35% and 45% are not effective in inhibiting the growth of Methicillin Resistant Staphylococcus aureus (MRSA) bacteria, however, the average zone of inhibition for spray preparations with ethanol extract of lemongrass leaves with a concentration of 45% is against MRSA bacteria. 11 mm is included in the strong category. Keywords: Lemongrass Leaves, MRSA Bacteria, Spray preparation.

Mec Genes in Methicillin Resistant Staphylococcus aureus

Methicillin-resistant Staphylococcus aureus (MRSA) manifests antibiotic resistance, especially for the β-lactams antibiotic group. MRSA bacteria are a common cause of infection in humans. The antibiotic resistance characteristic comes from the mecA, mecB, and mecC genes in the bacterial chromosome. mecA is the most common gene found in MRSA. Therefore, it is essential to know the role of the mecA gene in antibiotic resistance. This paper searched the literature about MRSA bacteria, the mec gene, and their relationship to cause resistance. The results showed that the mec gene found in MRSA bacteria causes antibiotic resistance in penicillin groups. Methicillin-resistant Staphylococcus aureus (MRSA) manifests antibiotic resistance, especially for the β-lactams antibiotic group. MRSA bacteria are a common cause of infection in humans. The antibiotic resistance characteristic comes from the mecA, mecB, and mecC genes in the bacterial chromosome. mecA is the most common gene found in MRSA. Therefore, it is essential to know the role of the mecA gene in antibiotic resistance. This paper searched literature about MRSA bacteria, the mec gene, and their relationship to cause resistance. The results showed that the mec gene found in MRSA bacteria causes antibiotic resistance in penicillin groups.

COMBATTING METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS (MRSA) IN THE FOOD INDUSTRY BY HARNESSING THE POWER OF NATURE: A SYSTEMATIC REVIEW

Antibiotic resistance is a critical global health concern, with Methicillin-resistant Staphylococcus aureus (MRSA) posing a significant challenge due to its resistance to commonly used antibiotics. Recent research has revealed the potential of natural compounds and microorganisms in combatting MRSA and other antibiotic-resistant bacteria. In this systematic review, we studied the effect of essential oils, bacteriophages, bacteriocins, and probiotics on S. aureus, including MRSA in particular, in the food industry. Essential oils (EOs) have gained significant attention because of their antimicrobial properties, inhibiting MRSA growth by damaging bacterial cells and inhibiting essential enzymes and compounds. Cinnamon oil liposomes caused the most significant decrease in MRSA populations among our reviewed essential oils. Bacteriophages can lyse the bacterial host. They encode peptidoglycan hydrolases called endolysins that target the bacterial cell wall. In our study, S. aureus phage (containing CHAPLysGH15 and LysGH15), and phage SA11 endolysin (LysSA11) were the most effective against S. aureus. Bacteriocins, antimicrobial peptides produced by bacteria, also show potential in combatting MRSA, mainly by generating organic acids that interfere with bacterial metabolism. According to our review, the most effective bacteriocins against S. aureus were Enterocin AS-48 with phenolic compounds or with 2NPOH, Bacteriocin isolated from Lactobacillus pentosus - Pentocin JL-1, and bacteriocin produced by S. pasteuri RSP-1, respectively. Probiotics can compete with pathogens by producing antimicrobial compounds that disrupt MRSA cell production and ultimately lead to bacterial death. In our review, the most effective probiotics were Streptomyces griseus, Pediococcus acidilactici strains A11 and C12, Lactococcus lactis, and Lactobionic acid respectively. A multi-hurdle approach combining these natural agents has shown promising results in targeting and eliminating MRSA cells. By harnessing the power of nature, we can potentially overcome the challenges posed by MRSA and other antibiotic-resistant bacteria.

Antimicrobial Effects of Propolis Ethanol Extract Tetragonula laeviceps Against Bacteria Methicillin Resistant Staphylococcus aureus

Staphylococcus aureus bacteria, especially Methicillin Resistant Staphylococcus aureus (MRSA), is one of the main causes of antibiotic resistance. MRSA is a bacteria that is resistant to the ?-lactam class of penicillin antibiotics. MRSA infections can cause a variety of health problems, including skin infections, pneumonia, and bacteremia. This problem requires alternative resources that can be an option. Propolis from the stingless bee species Tetragonula laeviceps can be a potential alternative for treating MRSA infections. This research is classified as a true experiment with a posttest control group design. The concentration of propolis ethanol extract tested was 20%; 10%; 5%; 2.5%; 1.25%; and 0.625% in the diffusion and dilution methods. These two methods provide assessment output in the form of inhibition zone diameter, minimum inhibition concentration, and minimum bactericidal concentration. The data obtained will be tested statistically using One-way Anova and Fisher's exact tests. Test results show that the ethanol extract of Tetragonula laeviceps propolis has an antimicrobial effect against MRSA bacteria. The diffusion test showed that the average diameter of the inhibition zone formed by the ethanol extract of propolis successively from a concentration of 20% to 1,25% with six repetitions was 12.7 ± 1.23 mm; 11.36 ± 0.67 mm; 10.24 ± 0.61 mm; 7.59 ± 1.69 mm; and 6.5 ± 1.14 mm. From these averages, the One-Way Anova test revealed a statistically significant difference in the diameter of the inhibition zone (F(7,40) = [ 192.79], p = 0.000).

Detection of Methicillin-Resistant Staphylococcus Aureus using vancomycin conjugated silica-based fluorescent nanoprobe

Methicillin-Resistant Staphylococcus Aureus (MRSA) is a worldwide major pathogenic bacteria that has emerged over the past three decades as the leading cause of nosocomial and community-acquired infections. Biosensors can provide rapid, sensitive, and selective detection of the presence and number of bacteria in various environments. Herein, a novel fluorescence nanoprobe was designed as a biosensor for MRSA detection using dye-incorporated silica nanoparticles (FSiNP). Based on the results of specific surface area analysis using the Brauner Emmett-Teller (BET) method, the surface area of the nanoparticles was obtained at 377.127 m2/g, and the X-ray diffraction (XRD) analysis confirmed that it was in the amorphous phase. Vancomycin, as the bioreceptor, was immobilized on the silica surface through a hydrosilylation reaction, generating the biosensing platform FSiNP-Van. Each modification step was corroborated by the Fourier Transform Infra-Red (FTIR) spectroscopy. The sensing principle was based on the fluorescence-quenching mechanism of FSiNP-Van at 515 nm obtaining a rapid response time of 20 min. The FSiNP-Van nanoprobe provided a wide linear concentration range of 10–106 CFU/mL with a limit of MRSA detection calculated at 1 CFU/mL. The fluorescent nanoprobe demonstrated here is expected to find applications in point-of-care (POC) diagnostics to detect the presence of MRSA bacteria.

Bimetallic oxide Cu–Fe3O4 nanoclusters with multiple enzymatic activities for wound infection treatment and wound healing

Infections and oxidative stress complicate wound healing. In recent years, nanomaterials with natural enzymatic activities have enabled the development of new antibacterial pathways. In this study, Cu–Fe3O4 nanoclusters with multienzyme properties were synthesized. Interestingly, they exhibited activity similar to that of horseradish peroxidase (POD) in acidic environments but their functions resembled superoxide dismutase and catalase in neutral or weakly alkaline environments. In vitro studies have demonstrated the good free-radical scavenging activity of Cu–Fe3O4 nanoclusters in a neutral environment. Under acidic conditions, Cu–Fe3O4 nanoclusters combined with H2O2 demonstrated good antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli). The combination of Cu–Fe3O4 and H2O2 was found to be effective in preventing MRSA infections and promoting wound healing in animal models. RNA sequencing (RNA-seq) technology revealed that chemodynamic therapy (CDT) using nanoparticles can interfere with metabolic processes such as galactose metabolism in MRSA bacteria, destroy the transport system on the surface of MRSA, and affect quorum sensing to hinder the formation of biofilms, thus achieving effective antibacterial efficacy. The use of Cu–Fe3O4 nanoclusters as a novel class of multi-catalytically active nanozymes in the anti-infection of disease-causing pathogens and wound healing has significant potential. Statement of significanceCu–Fe3O4 nanoclusters with multienzyme properties were successfully prepared by a solvothermal method. Cu–Fe3O4 nanoclusters exhibited horseradish peroxidase (POD)-like activity in acidic environments and also showed synergistic effects similar to superoxide dismutase peroxidase in neutral or weakly basic environments. More importantly, these Cu–Fe3O4 nanoclusters showed high biosafety with no apparent in vivo toxicity. Chemodynamic therapy (CDT) using Cu–Fe3O4 nanoclusters was revealed by RNA sequencing (RNA-Seq) technology to interfere with the metabolic processes of MRSA bacteria, such as galactose metabolism, disrupt the MRSA surface transport system, and impede biofilm formation, resulting in effective antibacterial efficacy. The use of Cu–Fe3O4 nanoclusters for anti-infection and wound healing of pathogenic pathogens has significant potential as a novel class of multi-catalytic active nanoclusters.

Antibacterial and antibiofilm activity of Abroma augusta stabilized silver (Ag) nanoparticles against drug-resistant clinical pathogens.

Infectious diseases remain among the most pressing concerns for human health. This issue has grown even more complex with the emergence of multidrug-resistant (MDR) bacteria. To address bacterial infections, nanoparticles have emerged as a promising avenue, offering the potential to target bacteria at multiple levels and effectively eliminate them. In this study, silver nanoparticles (AA-AgNPs) were synthesized using the leaf extract of a medicinal plant, Abroma augusta. The synthesis method is straightforward, safe, cost-effective, and environment friendly, utilizing the leaf extract of this Ayurvedic herb. The UV-vis absorbance peak at 424nm indicated the formation of AA-AgNPs, with the involvement of numerous functional groups in the synthesis and stabilization of the particles. AA-AgNPs exhibited robust antibacterial and antibiofilm activities against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE). The MIC values of AA-AgNPs ranged from 8 to 32μg/mL. Electron microscopic examination of the interaction of AA-AgNPs with the test bacterial pathogens showed a deleterious impact on bacterial morphology, resulting from membrane rupture and leakage of intracellular components. AA-AgNPs also demonstrated a dose-dependent effect in curtailing biofilm formation below inhibitory doses. Overall, this study highlights the potential of AA-AgNPs in the successful inhibition of both the growth and biofilms of MRSA and VRE bacteria. Following studies on toxicity and dose optimization, such AgNPs could be developed into effective medical remedies against infections.

Synergistic effect of commercial mangosteen extract (Garcinia mangostana L.) and amoxicillin against methicillin-resistant Staphylococcus aureus (MRSA)

Antibiotic resistance occurs worldwide, and has become a threat to humankind. Previous data have shown that antimicrobial resistance is a global issue demanding immediate resolution because it threatens the environment and society. The present work thus investigated the synergistic effects of commercial Garcinia mangostana L. (GML) extract and amoxicillin on the growth of methicillin-resistant Staphylococcus aureus (MRSA) bacterial cells. A commercial GML extract was screened for phytochemical properties, and the presence of α-mangostin was detected using high-performance liquid chromatography (HPLC). The antibacterial activity of the commercial GML extract with amoxicillin was analysed by minimum inhibitory concentration (MIC) and checkerboard assays. The morphology ultrastructure of bacteria was observed using transmission electron microscopy (TEM), after treatment with commercial GML extract, either single or in combination with amoxicillin. The MICs of amoxicillin and commercial GML extract against MRSA bacteria were 250.00 and 137.50 μg/mL, respectively. The checkerboard assay showed synergistic activity in the combination of commercial GML extract (34.38 µg/mL) and amoxicillin (62.50 µg/mL) at fractional inhibitory concentration (FIC) index of < 0.5. Damage to the structure of bacteria occurred due to the commercial GML extract plus amoxicillin. It was observed that the loss of bacterial cell membranes led to an irregular bacterial structure. These findings provided evidence that the combination of commercial GML extract and amoxicillin could reverse bacterial resistance in order to determine the susceptibility of traditional drugs.

Detection of Methicillin-Resistant Staphylococcus Aureus Bacteria Using Liquid Crystals

Methicillin-resistant Staphylococcus aureus (MRSA) is an essential pathogen for public health and this bacteria commonly cause serious infectious in humans. In recent years, MRSA bacteria are detected by the bacterial culture and nucleic acid-based methods which are time-consuming and labor-intensive. In this study, a novel liquid crystal (LC)-based biosensing system was developed to overcome these limitations. The objective of this study was to detect the presence of MRSA bacteria which prepared within the isotonic water and phosphate buffer saline (PBS). In this system, the binding of MRSA bacteria to the dimethyloctadecyl [3-(trimethoxysilyl) propyl] ammonium chloride (DMOAP)-coated surface disrupted the orientation of LCs, triggering a transition from a homeotropic orientation to a random one. This transition in the orientation of the LCs was observed as a change from a dark optical LC image to a bright one under a polarized light microscope (POM) and the reflection values of LC molecules were determined by using a spectrometer. Through this sensing mechanism, MRSA bacteria prepared within the isotonic water was detected ranging from the 9.2x103 CFU/mL to 9.2x107 CFU/mL concentration. Furthermore, MRSA bacteria prepared in PBS was detected in the concentration range of 7.1x104 CFU/mL to 7.1x108 CFU/mL by using this system.

IN VITRO ANTIBACTERIAL ACTIVITY OF EUCALYPTUS (Melaleuca leucadendra) OIL AGAINST METHICILLIN-RESISTANT Staphylococcus aureus (MRSA)

Highlights The use of eucalyptus oil is natural and risk-free. It has a lengthy history of use as a traditional medicine in Asia, including Indonesia. It has been demonstrated that eucalyptus oil possesses antibacterial activity against MRSA, and this activity was controlled by the oil's concentration. Abstract Background: Eucalyptus (Melaleuca leucadendra) oil is used by Indonesians as an herbal medicine. Eucalyptus containing 1,8-cineol at 72.30% is expected to be used as an antibacterial. Objective: The study aimed to assess the antibacterial activity of eucalyptus oil against Methicillin-resistant Staphylococcus aureus (MRSA) bacteria in vitro. Material and Method: The materials used were eucalyptus oil (M. leucadendra) in various concentrations with ethyl acetate as solvent. The research method used was the agar-well diffusion assay. The MRSA was suspended to 0.5 McFarland turbidity. The MRSA suspension was thoroughly swabbed onto the surface of the Mueller-Hinton agar plate. The wells were made with a diameter of 0.6 mm on Muller-Hinton agar aseptically and 100 µl of eucalyptus oil was put into the well using a micropipette and incubated for 24 hours at 37°C. The diameter of the inhibition zone was measured with a caliper. Statistical analysis using the SPSS software edition 23. Result: Eucalyptus oil solution starting from a concentration of 10% to 100% had an antibacterial response, which could be seen by the formation of a bacterial inhibition zone around the eucalyptus oil (M. leucadendra) wells. The inhibition ability of eucalyptus oil against MRSA bacteria was greatest at a concentration of 90% with an inhibition zone of 31.26 mm. It was found that the concentration of eucalyptus oil affected the diameter of the bacterial inhibition area. Conclusion: Eucalyptus oil has been shown to have antibacterial activity against MRSA and was influenced by the concentration of the oil.

Antimicrobial effect of areca nut ethanol extract against methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA)

Background: Developing herbal medicines for antibacterial activity has been challenging in Indonesia. Of the alternative antimicrobials, areca nut (Areca catechu L) has antimicrobial activity since it contains flavonoids, tannins, phenols, alkaloids, saponins, terpenoids, and glycosides. The use of antibiotics in underdeveloped and developing countries is still irrational, and bacterial infection in Indonesia is still a problem to resolve, so searching for new herbal medicines is very important. Staphylococcus aureus is the most common normal flora that causes infection. The two types of Staphylococcus aureus are Methicillin-resistant Staphylococcus aureus (MRSA) and Methicillin-sensitive Staphylococcus aureus (MSSA). The study aimed to determine the efficacy of the antibacterial activity of areca nut ethanol extract against MRSA and MSSA isolates.
 Methods: The research was conducted at the Biomedical Laboratory, Faculty of Medicine and Health Sciences, University of Jambi. Young and old areca nut ethanol extracts were macerated using 96% ethanol. The antibacterial activity test of the ethanol extract was carried out using the Kirby-Bauer disc diffusion method against the isolates of MRSA (clinical isolates) and MSSA (SA 25923/K+, SA 29213/K-).
 Results: The results showed that the phytochemicals of young areca nut extracts contained alkaloids, flavonoids, phenols, and steroids, and old areca nut extracts contained alkaloids, flavonoids, and phenols. The MRSA isolates did not show the activity of the two areca nut ethanol extracts and were also resistant to Ceftadizime, Gentamicin, Ceftriaxone, and Cefoxitin. The MSSA isolates showed strong activity against old areca nut ethanol extract, moderate activity against young areca nut ethanol extract, and resistance to gentamicin. 
 Conclusion: MRSA bacteria are resistant to the ethanol extract of young and old areca nuts, while the ethanol extract of old areca nuts has strong antibacterial activity against MSSA. 
 Keywords: antibacterial, areca seed ethanol extract, MRSA, MSSA

Antibacterial Activity of Honey Pineapple Peel Extract (Ananas comosus [L] Merr.) against MethicillinResistant Staphylococcus aureus

Methicillin-resistant Staphylococcus aureus (MRSA) infection is a global health problem that needs to be answered by developing a synthetic or natural antibacterial to overcome the problem. The honey pineapple peel contains potentially antibacterial compounds such as bromelain and flavonoid. This study aims to determine the antibacterial activity of honey pineapple peel extract (Ananas comosus [L] Merr.) against MRSA. The design of this study was a true experimental study with a post-test control group design. This study examined the antibacterial activity of honey pineapple peel extract at concentrations of 100%, 50%, 25%, 12.5%, 6.25%, 3.125%, and 1.56% against MRSA bacteria by measuring minimum inhibitory concentration (MIC) with dilution method and minimum bactericidal concentration (MBC). The results of this study show that MIC was obtained at concentrations of 50 % and MBC at extract with a concentration of 100 %. There is an antibacterial activity of honey pineapple peel extract (Ananas comosus [L] Merr.) against MRSA.
 Keywords: Antibacterial Activity, Honey Pineapple Peel, MRSA

A novel rodent model of chronic spinal implant-associated infection

BACKGROUND CONTEXTBacterial infection of spinal instrumentation is a significant challenge in spinal fusion surgery. Although the intraoperative local application of powdered vancomycin is common practice for mitigating infection, the antimicrobial effects of this route of administration are short-lived. Therefore, novel antibiotic-loaded bone grafts as well as a reliable animal model to permit the testing of such therapies are needed to improve the efficacy of infection reduction practices in spinal fusion surgery. PURPOSEThis study aims to establish a clinically relevant rat model of spinal implant-associated infection to permit the evaluation of antimicrobial bone graft materials used in spinal fusion. STUDY DESIGNRodent study of chronic spinal implant-associated infection. METHODSInstrumentation anchored in and spanning the vertebral bodies of L4 and L5 was inoculated with bioluminescent methicillin-resistant Staphylococcus aureus bacteria (MRSA). Infection was monitored using an in vivo imaging system (IVIS) for 8 weeks. Spines were harvested and evaluated histologically, and colony-forming units (CFUs) were quantified in harvested implants and spinal tissue. RESULTSPostsurgical analysis of bacterial infection in vivo demonstrated stratification between MRSA and phosphate-buffered saline (PBS) control groups during the first 4 weeks of the 8-week infection period, indicating the successful establishment of acute infection. Over the 8-week chronic infection period, groups inoculated with 1 × 105 MRSA CFU and 1 × 106 MRSA CFU demonstrated significantly higher bioluminescence than groups inoculated with PBS control (p = 0.009 and p = 0.041 respectively). Histological examination at 8 weeks postimplantation revealed the presence of abscesses localized to implant placement in all MRSA inoculation groups, with the most pervasive abscess formation in samples inoculated with 1 × 105 MRSA CFU and 1 × 106 MRSA CFU. Quantification of CFU plated from harvested spinal tissue at 8 weeks post-implantation revealed the 1 × 105 MRSA CFU inoculation group as the only group with a significantly greater average CFU count compared to PBS control (p = 0.017). Further, CFU quantification from harvested spinal tissue was greater than CFU quantification from harvested implants across all inoculation groups. CONCLUSIONOur model demonstrated that the inoculation dosage of 1 × 105 MRSA CFU exhibited the most robust chronic infection within instrumented vertebral bodies. This dosage had the greatest difference in bioluminescence signal from control (p < 0.01), the lowest mortality (0% compared to 50% for samples inoculated with 1 × 106 MRSA CFU), and a significantly higher amount of CFUs from harvested spine samples than CFUs from control harvested spine samples. Further, histological analysis confirmed the reliability of this novel rodent model of implanted-associated infection to establish infection and biofilm formation of MRSA for all inoculation groups. CLINICAL SIGNIFICANCEThis model is intended to simulate the infection of instrumentation used in spinal fusion surgeries concerning implant locality and material. This model may evaluate potential antimicrobial and osteogenic biomaterials and investigate the relationship between implant-associated infection and failed fusion.

Synergistic Inhibition of Methicillin-Resistant Staphylococcus aureus (MRSA) by Melaleuca alternifolia Chell (Tea Tree) and Eucalyptus globulus Labill. Essential Oils in Association with Oxacillin.

The presence of antibiotic-resistant bacteria has become a major therapeutic priority. This trend indicates the need for alternative agents to antibiotics, such as natural compounds of plant origin. By assessing membrane permeability, we investigated the antimicrobial activity of Melaleuca alternifolia and Eucalyptus globulus essential oils (EOs) against three strains of methicillin-resistant Staphylococcus aureus (MRSA). Using the checkerboard method, the efficacy of single EOs, in association with each other or in combination with oxacillin, was quantified by calculating the fractional inhibitory concentrations (FIC Index). All EOs showed a reduction in bacterial load, an alteration of membrane permeability which leads to an increase in its function, resulting in the release of nucleic acids and proteins. The treatment with EO-oxacillin combinations and associated EO-EO resulted in a synergistic effect in most of the tests performed. EO-EO association showed a high activity in the alteration of the membrane, increasing the permeability to about 80% in all the MRSA strains treated. In conclusion, the combination of EOs and antibiotics represents a valid therapeutic support against MRSA bacteria, allowing for a decrease in the antibiotic concentration needed for therapeutic use.

Effectiveness of ozone-laser photodynamic combination therapy for healing wounds infected with methicillin-resistant Staphylococcus aureus in mice.

According to 2013 data from the Ministry of Health of the Republic of Indonesia, there were 8.2% more wounds than typical in Indonesia; 25.4% were open wounds, 70.9% were abrasions and bruises, and 23.2% were lacerations. A wound is defined as damage or loss of body tissue. This study aimed to determine the effectiveness of wound healing using red-laser therapy (650 nm, 3.5 J/cm2), blue-laser therapy (405 nm, 3.5 J/cm2), ozone therapy, red-laser therapy (650 nm, 3.5 J/cm2) with ozone, and blue-laser therapy (405 nm, 3.5 J/cm2) with ozone. One hundred and twelve mice were given incision wounds and infected with methicillin-resistant Staphylococcus aureus (MRSA). The study used a factorial design with two factors: The type of therapy (n = 7) and irradiation time (days 1, 2, 4, and 6). The mice were divided into seven therapy groups: Control group with NaCl, control with Sofra-tulle® treatment, red-laser therapy (650 nm, 3.5 J/cm2), blue-laser therapy (405 nm, 3.5 J/cm2), ozone therapy, red-laser therapy (650 nm, 3.5 J/cm2) with ozone, and blue-laser therapy (405 nm, 3.5 J/cm2) with ozone. This therapy was performed using irradiation perpendicular to the wound area. The photosensitizer used was curcumin 10 mg/mL, which was applied to the wound area before exposure to a laser and ozone. The ozone concentration was 0.011 mg/L with a flow time of 80 s. The test parameters were the number of collagens, bacterial colonies, lymphocytes, monocytes, and wound length measurement to determine their acceleration effects on wound healing. Data were analyzed by a two-way (factorial) analysis of variance test. Acceleration of wound healing was significantly different between treatments with a laser or a laser-ozone combination and treatment using 95% sodium chloride (NaCl) and Sofra-tulle®. On day 6, the blue-laser with ozone treatment group had efficiently increased the number of bacteria and reduced the wound length, and the red-laser treatment with ozone increased the amount of collagen. In addition, the red-laser also reduced the number of lymphocytes and monocytes, which can have an impact on accelerating wound healing. Blue-laser therapy was very effective for increasing the number of epithelia. The blue- and red-laser combined with ozone treatments effectively accelerated the healing of incisional wounds infected with MRSA bacteria.

Growth Inhibitory Effects of Red and Yellow Passion Fruits against MRSA and ESBL-producing Bacteria

Background: Red passion fruit (Passiflora edulis Sims) and yellow passion fruit (Passiflora edulis f. flavicarpa) are native Indonesian fruits with numerous health benefits. This study used de Man, Rogosa, and Sharpe (MRS) medium for fermentation. Probiotics are beneficial microorganisms obtained from fermented food or drink (Hamid et al., 2020). Probiotics and lactic acid bacteria, which are known to have various benefits, particularly as antibacterial, are among the active components identified in passion fruit pulp. Objective: This study examinedthe antibacterial activity of red and yellow passion fruits. Methods: Freshly collected passion fruit pulps were fermented in MRS medium in a shaker incubator for 48 hours at 24°C. The filtrates from the fermented broth were tested against clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and extended-spectrum beta-lactamase (ESBL)-producing bacteria. The analyses applied the streak plate method and the total plate count method. Results: The Minimum Inhibitory Concentrations (MICs) of red passion fruit ferment filtrate against MRSA 10 and MRSA 11 were 50% and 60%, respectively, and of yellow passion fruit ferment filtrate against MRSA 10 and MRSA 11 were both 30%. Meanwhile, the MICs of red passion fruit ferment filtrates were 35% against ESBL 41 and ESBL 43 and 25% against ESBL 45 and ESBL 47, whereas the MICs of yellow passion fruit (Passiflora edulis f. favicarpa) ferment filtrates were 25% against ESBL 41 and ESBL 43, and 12.5% against ESBL 45 and ESBL 47. Red passion fruit (Passiflora edulis Sims) and yellow passion fruit (Passiflora edulis f. flavicarpa) ferment filtrates became growth inhibitors against the clinical isolates of MRSA and ESBL-producing bacteria with an optimal fermentation time of 24 hours and an optimal concentration of 75%. Conclusion: The results of this study found that the fermented filtrates of red and yellow passion fruits in MRS media could be developed as an antibacterial against MRSA and ESBL-producing bacteria.