Methicillin-resistant Staphylococcus aureus (MRSA) is a serious threat to the global healthcare system. Ineffective and resistance to antibiotic treatments have increased morbidity and mortality rates worldwide.New and effective antibiotics are needed to combat against bacterial resistance. Endophytic fungi are crucial reservoirs of novel bioactive metabolites. In particular, the secondary metabolites show promising therapeutic potential, notably, antibacterial. This review discussed the emerging potential of endophytic fungi as anti-MRSA agents. The ecological sources of endophytic fungi were discussed with the synthesis of bioactive metabolites.The mode of antibacterial actions was elucidated to give a better understanding of the mechanisms involved. This review may serve as an important reference for future discovery and developments of anti-MRSA agents from endophytic fungi.

Aims: To evaluate the antibacterial efficacy of ethyl acetate extract of Aspergillus flavus IBRL-C8 against Gram-positive and Gram-negative bacteria. Methodology and results: In this experiment, an endophytic fungus which identified as A. flavus IBRL-C8 was extracted using ethyl acetate and methanol, from Senna siamea, prior to in vitro antibacterial test on eight Gram-bacteria. The results were significantly more enunciated to the ethyl acetate extract since the Gram-bacteria signified 9.0 to 20.0 mm of inhibition zones on Muller Hinton Agar (MHA) during disc diffusion assay. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the extract were ranged from 125-1000 µg/mL and 125-2000 µg/mL, respectively. Time-kill assay depicted the ethyl acetate extract of A. flavus IBRL-C8 exceptionally retarded methicillin-resistant Staphylococcus aureus (MRSA) and also manifested extended antibacterial activity. The maximum reduction in cell numbers occurred at 2MIC concentration (250 µg/mL) during the interval time of 16 h. The malformations noticed from microscopic observations where the transformation of structural annihilation from regular spherical morphology to non-spherical shape with an irregular surface and also disruption around the cell membrane when the MRSA treated with ethyl acetate extract of A. flavus IBRL-C8. Conclusion, significance and impact of study This study proposed the ethyl acetate extract of A. flavus IBRL-C8 as a potential antibacterial agent against MRSA infection, which can be useful in pharmaceutical application.
Aspergillus flavus ; Anti-Bacterial Agents

Aims: This research was conducted to develop and characterize polyvinyl alcohol (PVA)/montmorillonite (MONT) clay incorporated with carvacrol (Carva) nanocomposite film as a potential material in wound dressing. Methodology and results: Organophilic MONT clay, which was initially modified from commercial MONT clay by cetyltrimethylammonium bromide (CTAB), was used in the polymerization process using PVA. The synthesized nanocomposites were visualized via transmission electron microscopy (TEM). The developed film (PVA/MONT/Carva nanocomposite film) was characterized via Fourier transform infrared (FTIR). The investigation on mechanical property and antimicrobial activity of the film was also performed. All nanocomposites are spherical, with a size of 92.8 ± 22.1 nm. The -OH stretch, C-H stretch, aromatic group, SiO stretch, and C-O from acetyl group were identified in the PVA/MONT/Carva nanocomposite films. During the chemical release test, carvacrol attained a plateau at 24 h, with a total release of 62.3%. This nanocomposite exhibited a severe detrimental influence on the growth of Gram-bacteria and yeasts, which represented a broad spectrum of antimicrobial agents. All test microorganisms showed approximately up to 82% reduction of microbial growth during the Hohenstein challenge test. Physically, the nanocomposite films were yellowish and apparent. The film was sturdy, flexible, elastic and consisted of excellent water holding capacity. Conclusion, significance and impact of study PVA/MONT/Carva nanocomposite film may have a useful potential to be merged in the pharmaceutical application, especially in wound dressing production.
Polyvinyl Alcohol ; Bentonite ; Wound Healing

Aims: This present study focused on purification of fungal β-mannanase produced by Aspergillus niger USM F4 and also physicochemical characterisation of the purified enzyme. Methodology and results: The purified β-mannanase with a molecular mass of ~47.4 kDa was demonstrated on SDSPAGE gel. The enzyme signified a purification degree of 4-fold, with final specific activity of 196.42 U/mg. It reached an optimum catalytic activity at pH 4.0 and 60 °C. The thermal stability of the enzyme was up to 70 °C and maintained the 50% activity after 30 min at 80 °C. Meanwhile, the pH stability was in the range of pH 3.0-9.0 and a 30 min half-life at pH 10.0. All chemical substances manifested an inhibitory effect on purified β-mannanase, with SDS (28.16 ± 0.05% residual activity) as the strongest inhibitor, followed by cupric ion (Cu2+) (49.51 ± 0.09% residual activity). As a whole, the enzyme displayed a substrate specificity in the order of locust bean gum (LBG) > carboxymethylcellulose > soluble starch > xylan from oat spelt > α-cellulose. Its preference for LBG has generated the Km and Vmax values of 0.20 mg/mL and 9.82 U/mL, respectively. Conclusion, significance and impact of study The outcomes of our study offer potential for use at industrial scales, particularly in the oligosaccharides production that involve acid-related activity, wide-ranging temperature and pH stability.
Aspergillus niger ; beta-Mannosidase

Aims: Endophytic fungi are organisms that colonize living plant tissues without causing significant symptoms of diseases. They are known as the repository of bioactive compounds. This study was aimed to isolate endophytic fungi associated with Aloe vera and to assess their antimicrobial activity against the diabetic wound pathogens that result to lower limb amputations. Methodology and results: In this study, 84 endophytic fungi were isolated from A. vera by using 5% sodium hypochlorite and 70% ethanol as surface sterilants. On preliminary antimicrobial screening by agar plug assay, 12 (14.3%) isolates showed significant antagonistic activity on the test microorganisms. Two of the endophytes were identified as Aspergillus flavus and Colletotrichum gloeosporioides through morphological and molecular techniques. The fungal cultures fermented in shake flask were extracted subsequently with hexane, dichloromethane, ethyl acetate, and n-butanol. Ethyl acetate and dichloromethane extracts of A. flavus and C. gloeosporioides inhibited all test bacteria. The compounds in the extracts were seemed to be semi-polar in nature. On broth microdilution assay, minimal inhibitory and minimal lethality concentration of ethyl acetate extracts of the fungi were found in the range of 0.63−2.50 and 1.25−5.00 mg/mL, respectively. Conclusion, significance and impact of study This study reveals that A. vera harbours a wide diversity of endophytic fungi. A. flavus and C. gloeosporioides showing broad spectrum antimicrobial activity on wound pathogens may be potential sources of bioactive compounds. Further investigations should be conducted to isolate and identify the antimicrobial compound produced by these fungal isolates.
Endophytes--isolation & ; purification ; Aloe

Aims: Diabetic patients with foot ulcer showed 150-fold increased risk of amputation, which is primarily caused by microbial infection. Silver ions are commonly incorporated into wound dressing to enhance the antimicrobial property. However, concerns have been expressed about the development of bacterial resistance to heavy metals. In this study, we extracted the nano-cellulose from medical cotton and reinforced with gelatin to develop a film for wound dressing. Methodology and results: Garcinia mangostana L pericarp extract was incorporated into the nano-cellulose film as antimicrobial finishing. The efficacy of the developed nano-cellulose film was evaluated on diabetic wound microorganisms. We observed cellulose nano crystals with an average length of 133.71 nm under transmission electron microscope. The developed film showed gradual release of the extract over a period of 48 h and no burst effect was observed. The film exhibited significant inhibitory activity on three Gram positive bacteria, three Gram negative and all filamentous fungi tested. On Hohenstein challenge test, all test microorganisms showed significant growth reduction, with the treatment of the film. We also noticed that the antimicrobial activity of the film sustained even after 20 washes. Conclusion, significance and impact of study Our results indicate that the G. mangostana L pericarp extract loaded nano-cellulose films exhibited significant inhibitory activity on diabetic wound microorganisms. The developed film can be potentially used to prevent foot ulcer infection among diabetic patients.

Microbial wound infection prolonged the hospitalization and increase the cost for wound management. Silver is commonly used as antimicrobial wound dressing. However, it causes several adverse side effects. Hence, this study was aimed to evaluate the antimicrobial efficiency of Swietenia macrophylla seed extract on clinical wound pathogens. Besides, the bioactive constituents of the seed extract were also determined. S. macrophylla seeds were extracted with methanol by maceration method. The seed extract inhibited 5 test bacteria and 1 yeast on disc diffusion assay. The antibacterial activity was broad spectrum, as the extract inhibited both Gram positive and Gram negative bacteria. On kill curve analysis, the antibacterial activity of the seed extract was concentration-dependent, the increase of extract concentration resulted in more reduction of bacterial growth. The extract also caused 99.9% growth reduction of Bacillus subtilis relative to control. A total of 21 compounds were detected in gas chromatography-mass spectrometry analysis. The predominant compounds present in the extract were oleic acid (18.56%) and linoleic acid (17.72%). In conclusion, the methanolic extract of S. macrophylla seeds exhibited significant antimicrobial activity on clinical wound pathogens. Further investigations should be conducted to purify other bioactive compounds from the seeds of S. macrophylla.
Bacillus subtilis ; Bacteria ; Bandages ; Diffusion ; Gas Chromatography-Mass Spectrometry ; Gram-Negative Bacteria ; Hospitalization ; Linoleic Acid ; Meliaceae ; Methanol ; Methods ; Oleic Acid ; Silver ; Wound Infection ; Wounds and Injuries ; Yeasts

Aims: Food borne infections are routinely observed and frequently reported. Cymbopogon nardus, or commonly known as citronella is a tropical plant widely used for aromatherapy and insect repellant. This study aimed to evaluate the phytochemical compositions of C. nardus ethanol extract and its antimicrobial activity on food borne microorganisms. Methodology and results: The plant materials were extracted with ethanol using maceration technique. The ethanol extract obtained was analysed using GC-MS and the most abundant compounds were 6-methyloctahydrocoumarin (31.5%) and 2,4-di-tert-butylphenol (28.7%). On Kirby Bauer assay, the extract showed wide spectrum inhibitory effect on one Gram-positive bacteria (Bacillus coagulans), two Gram-negative bacteria (Proteus mirabilis and Escherichia coli), and one yeast (Candida albicans). All the test microorganisms showed different susceptibility to the extract, where the minimal inhibitory concentrations were ranged from 63 to 250 μg/mL. The inhibitory effect of the extract was bactericidal. In addition, the time kill analysis of the extract exhibited inhibitory effect on P. mirabilis which was concentration-dependent. The C. nardus extract exhibited rapid bactericidal effect on P. mirabilis and lower bacterial counts were obtained with increased extract concentration. Besides, the extract did not exert significant toxicity effect to Artemia salina on brine shrimp lethality assay. Conclusion, significance and impact of study Therefore C. nardus can be a potential source of safe and effective antimicrobial agent to combat food borne infections.

Aims: Endophytes are microorganisms residing in the living tissues of the host plant and may contribute to their hostplant by producing a plethora of bioactive compounds that provide survival value to the plant. This study aimed toevaluate the antimicrobial activity of Aspergillus sp. IBRL MP15 CCL, an endophytic fungus isolated from Swieteniamacrophylla leaf.Methodology and results: The antimicrobial activity was evaluated with disc diffusion and a colorimetric brothmicrodilution test against 15 organisms comprising of 4 Gram-positive bacteria and 4 Gram-negative bacteria, 4 fungiand 3 yeast. On disc diffusion assay, the fungal extract was shown to inhibit the growth of 7 test bacteria and 3 testyeast. The antibacterial activity was more pronounced with extract from fungal culture with host plant extractsupplementation with significantly larger inhibition zones on all susceptible test microorganisms. The minimal inhibitoryconcentration of the extract ranged from 250 to 4000 μg/mL indicating different level of susceptibility of the testedpathogens against the fungal extract. The killing kinetic study shows that antimicrobial activity of the fungal extract isconcentration dependent and it can act as bactericidal at higher concentration.Conclusion, significance and impact of study: The findings of this study suggest that Aspergillus sp. IBRL MP15CCL can be a promising source of antimicrobial agent to be further studied and developed

Helminthostachys zeylanica is a rare plant grows in lightly shaded areas. The fern was traditionally used as antipyretic and antiphlogistic agents. This study was aimed to evaluate the antibacterial potential of H. zeylanica on foodborne Bacillus cereus. The chemical composition of its ethanolic extract was also determined. The plant samples were collected at Kampung Kebun Relong, Kedah, Malaysia. The ethanolic extract showed significant inhibitory activity on B. cereus with a sizeable clear zone detected on disc diffusion assay. On broth microdilution assay, the MIC of the extract on B. cereus was 6.25 mg/ml and the MBC was 12.5 mg/ml. The inhibitory activity of the extract on B. cereus was bactericidal. In the growth dynamic study, the antibacterial efficacy of the extract was concentration dependent, where a lower colony forming unit count was obtained with increased extract concentration. The SEM micrograph of extract treated B. cereus cells showed invaginations of cell wall. The bacterial cell structure collapsed after 24 h exposure to the extract. The GCMS analysis of the extract showed that the major constituents of the extract were phenol (36.26%) and quercetin (29.70%). This study is important as it shows the potential use of H. zeylanica as an effective agent to control B. cereus related infections.
Bacillus cereus ; Bacillus ; Cell Wall ; Diffusion ; Ethanol ; Ferns ; Gas Chromatography-Mass Spectrometry ; Malaysia ; Phenol ; Plants ; Quercetin ; Stem Cells