Staphylococcus aureus is an important human pathogen in both hospital and the community that has demonstrated resistance to all currently available antibiotics over the last two decades. Multidrug-resistant isolates of methicillin-resistant S. aureus (MRSA) exhibiting decreased susceptibilities to glycopeptides has also emerged, representing a crucial challenge for antimicrobial therapy and infection control. The availability of complete whole-genome nucleotide sequence data of various strains of S. aureus presents an opportunity to explore novel compounds and their targets to address the challenges presented by antimicrobial drug resistance in this organism. Study compounds α-amyrin [3β-hydroxy-urs-12-en-3-ol (AM)], betulinic acid [3β-hydroxy-20(29)-lupaene-28-oic acid (BA)] and betulinaldehyde [3β-hydroxy-20(29)-lupen-28-al (BE)] belong to pentacyclic triterpenoids and were reported to exhibit antimicrobial activities against bacteria and fungi, including S. aureus. The MIC values of these compounds against a reference strain of methicillin-resistant S. aureus (MRSA) (ATCC 43300) ranged from 64 µg/ml to 512 µg/ml. However, the response mechanisms of S. aureus to these compounds are still poorly understood. The transcription profile of reference strain of MRSA treated with sub-inhibitory concentrations of the three compounds was determined using Affymetrix GeneChips. The findings showed that these compounds regulate multiple desirable targets in cell division, two-component system, ABC transporters, fatty acid biosynthesis, peptidoglycan biosynthesis, aminoacyl-tRNA synthetase, ribosome and β-lactam resistance pathways which could be further explored in the development of therapeutic agents for the treatment of S. aureus infections.
Entero- and exfoliative toxin gene profiling of 237 methicillin-susceptible Staphylococcus aureus (MSSA) isolated
from Universiti Kebangsaan Malaysia Medical Centre (UKMMC) were carried out via PCR amplification. Among
the tested toxin genes, sei was found to be the most prevalent (54.9%).
Skincare industries are growing rapidly around the globe but most products are formulated using synthetic chemicals and organic solvent extracted plant extracts, thus may be hazardous to the users and incur higher cost for purification that eventually leads to phytonutrient degradation. Therefore, this study aimed to formulate a stable natural formulation with antioxidant and antimicrobial activities by using supercritical carbon dioxide (SC-CO 2 ) extracted palm-pressed fiber oil (PPFO) as an active ingredient with virgin coconut oil (VCO) as a formulation base. PPFO was extracted from fresh palm-pressed fiber (PPF) while VCO was from dried grated coconut copra using SC-CO 2 before being subjected to the analyses of physicochemical properties, phytonutrient content and biological activities including antioxidant and antimicrobial. The nanoemulgel formulations were then developed and examined for their stability through accelerated stability study for 3 months by measuring their pH, particle size, polydispersity index and zeta potential. The results showed that PPFO contained a high amount of phytonutrients, especially total carotenoid (1497 ppm) and total tocopherol and tocotrienol (2269 ppm) contents. The newly developed nanoemulgels maintained their particles in nano size and showed good stability with high negative zeta potentials. Sample nanoemulgel formulated with 3% PPFO diluted in VCO as effective concentration showed significantly stronger antioxidant activity than the control which was formulated from 3% tocopheryl acetate diluted in mineral oil, towards DPPH and ABTS radicals, with IC 50 values of 67.41 and 44.28 µL/mL, respectively. For the antibacterial activities, the sample nanoemulgel was found to inhibit Gram positive bacteria S. aureus and S. epidermidis growth but not the Gram negative strain E. coli. Overall, this study revealed the potential of SF-extracted PPFO as an active ingredient in the antioxidant topical formulations thus future study on in vitro skin cell models is highly recommended for validation.
The synthesized titanium dioxide nanotubes (TiO2-NTs) were emerged as wound healing enhancer as well as exhibited significant wound healing activity on Sprague Dawley rats. In our present study, the blends of GG and TiO2-NTs bio-nanocomposite film was characterised by fourier transform infrared (FTIR), x-ray diffraction (XRD), scanning electron microscopy (SEM), thermogravimetric analysis, atomic force microscopy (AFM). The morphology of TiO2-NTs was investigated using transmission electron microscopy (TEM). The mechanical properties study shows that the GG + TiO2-NTs (20 w/w %) bio-nanocomposite film possessed the highest tensile strength and young modulus which are (4.56 ± 0.15) MPa and (68 ± 1.63) MPa, respectively. GG + TiO2-NTs (20 w/w %) also displays the highest antibacterial activity with (16 ± 0.06) mm, (16 ± 0.06) mm, (14 ± 0.06) mm, and (12 ± 0.25) mm inhibition zone were recorded against Staphylococcus aureus, Streptococcus, Escherichia coli, and Pseudomonas aeruginosa. The prepared bio-nanocomposite films have good biocompatibility against 3T3 mouse fibroblast cells and caused accelerated healing of open excision type wounds on Sprague Dawley rat model. The synergistic effects of bio-nanocomposite film like good swelling and WVTR properties, excellent hydrophilic nature, biocompatibility, wound appearance and wound closure rate through in vivo test makes it a suitable candidate for wound healing applications.
The spread of multidrug-resistant Staphylococcus aureus is a public health concern. The inducible macrolide-lincosamide-streptogrammin B (iMLSB ) phenotype (or inducible clindamycin resistance) is associated with false clindamycin susceptibility in routine laboratory testing and may lead to treatment failure. Tigecycline resistance remains rare in S. aureus worldwide. This study aims to determine the antimicrobial susceptibility profiles of clinical isolates of S. aureus obtained from the main tertiary hospital in Terengganu state, Malaysia, from July 2016 to June 2017. The antimicrobial susceptibilities of 90 methicillin-resistant S. aureus (MRSA) and 109 methicillin-susceptible S. aureus (MSSA) isolates were determined by disc diffusion with the iMLSB phenotype determined by D-test. Multidrug resistance (MDR) and the iMLSB phenotype were more prevalent in MRSA (84.4 and 46.7 %, respectively) compared to MSSA isolates. All five tigecycline-resistant isolates were MRSA. The high incidence of MDR and the iMLSB phenotype and the emergence of tigecycline resistance in the Terengganu S. aureus isolates warrants continuous vigilance.
Three new cembranoid diterpenes, 10-hydroxy-nephthenol acetate (1), 7,8-epoxy-10-hydroxy-nephthenol acetate (2), and 6-acetoxy-7,8-epoxy-10-hydroxy-nephthenol acetate (3), along with a known compound, 6-acetoxy-7,8-epoxy-nephthenol acetate (4), were isolated from the Bornean soft coral Nephthea sp. Antibacterial and anticancer activities were exhibited by compounds 1 and 2 against Staphylococcus aureus (ATCC 6538)/Escherichia coli (ATCC 13311) and Hela/MCF-7, respectively.
A thermostabilized, multiplex polymerase chain reaction (mPCR) assay was developed in this study for the detection of six respiratory bacterial pathogens. Specific primers were designed for an internal amplification control (IAC) and six target sequences from Klebsiella pneumoniae, Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa, Mycobacterium tuberculosis, and Haemophilus influenzae. The resultant seven-band positive amplification control (PAC) of this heptaplex PCR assay corresponded to 105 base pairs (bp) of IAC, 202 bp of K. pneumoniae, 293 bp of S. aureus, 349 bp of S. pneumoniae, 444 bp of P. aeruginosa, 505 bp of M. tuberculosis, and 582 bp of H. influenzae. Results found that 6% (w/v) of the stabilizer was optimum to preserve the functional conformation of Taq DNA polymerase enzyme. This assay was stable at ambient temperature for at least 6 months. The sensitivity and specificity of this assay were both 100% when testing on the intended target organisms (n = 119) and non-intended species (n = 57). The mPCR assay developed in this study enabled accurate, rapid, and simple detection of six respiratory bacteria.
Background: Foot infection is a major complication of diabetes mellitus (DM) and its agents are usually polymicrobial. This study aims to describe the agent and determine the association between polymicrobial infections and the severity of diabetic foot infections (DFI) and their outcomes.
Methods: This retrospective cohort study was conducted during one year and it involved 104 patients. Their records were reviewed and assessed. The causative agents and its sensitivity pattern were noted. The results were presented as descriptive statistic and analysed.
Results: A total of 133 microorganisms were isolated with 1.28 microorganisms per lesion. The microorganism isolated were 62% (n = 83) GN (Gram-negative) and 38% (n = 50) GP (Gram-positive). GN microorganisms include Pseudomonas spp (28%), Proteus spp (11%), Klebsiella spp (8%) and E. coli (4%). Staphylococcus aureus (54%) was predominant among GP, followed by Group B Streptococci (26%) and Enterococcus spp (6%). Thirty patients (28.8%) had polymicrobial infections. The association between the quantity of microorganisms and severity of DFI was significant. Among severe DFI cases, 77.8% with polymicrobial microorganisms underwent amputation compared to 33.3% with monomicrobial infection.
Conclusion: GN microorganisms were predominantly isolated from DFIs and remained sensitive to widely used agents. Polymicrobial infections were associated with DFI severity.
The incidence of methicillin-resistant Staphylococcus aureus (MRSA) infections in intensive care units in Malaysia is significant. Invasive MRSA infections are commonly treated with vancomycin. In clinical practice, the serum vancomycin trough concentration is used as a surrogate marker of vancomycin efficacy. A low concentration of vancomycin may result in less effective therapy and increase the risk of bacterial resistance. We evaluated the relationship between the resolution of MRSA infections and trough concentrations of vancomycin.
Background: A cross sectional study was carried out to determine the prevalence of microbiological contamination in ready- to-eat food and factors affecting it in Melaka from February 2008 till May 2008 in collaboration with the Food Safety and Quality Unit and District Health Offices in Melaka Tengah, Alor Gajah and Jasin.
Methods: A purposive sampling of 101 food premises was carried out and 202 ready- to-eat food samples were collected and analysed microbiologically using standard methods. A total of 202 food handlers were randomly selected for questionnaire interview while rectal and nasal swabs were taken from them to determine their healthy carrier status of pathogenic food bacteria.
Results: Results showed that the overall prevalence of microbiological contamination in ready- to-eat food was 35.1%, while prevalence by type of food was 42.0% for main dishes and 9.8% for staple food. The majority of the contaminations were due to coliforms (14.9%), total plate counts (12.4%) and Staphylococcus aureus (3.5%). The mean score for food premise inspection was 77.21 ± 10.32 and the prevalence of healthy carrier status of pathogenic food bacteria among the food handlers was 15.8%. The level of food handling practices among food handlers was influenced by sex (p=0.012) and formal training on food handling courses (p=0.009). There was a significant negative correlation between age and level of food handling practices among the food handlers (r = -0.163; p = 0.02). Poor food handling practices (p=0.02) and poor hygiene status among the food handlers (p
This study was conducted to detect the presence of heterogenous vancomycin-intermediate Staphylococcus aureus (heteroVISA) among MRSA isolates in a major hospital. Forty-three MRSA isolates with vancomycin MIC 2 microg/ml collected in 2009 was screened for heteroVISA using Etest Glycopeptide Resistance Detection (GRD) and confirmed by population analysis profile-area under curve method. The genetic relatedness of heteroVISA strains with other MRSA was examined by pulsed-field gel electrophoresis (PFGE) method. Two isolates were shown to be heteroVISA and derived from the same clone. This showed that heteroVISA strains were already present among our local strains since 2009 and were genetically related to other susceptible strains.
Commercial low-density polyethylene (LDPE) films were UV/ozone treated and coated using a layer-by-layer (LbL) technique by alternating the deposition of polyethyleneimine (PEI) and poly(acrylic acid) (PAA) polymer solutions and antimicrobial silver (Ag). The effects of the initial pH of the PEI/PAA polymer solutions alternating layers (pH 10.5/4 or 9/6.5) on the antimicrobial activity of the developed LbL coatings combined with Ag against Gram-negative and Gram-positive bacteria were investigated. The results from fourier transform infrared spectroscopy and toluidine blue O assay showed that LDPE LbL coated using PEI/PAA polymer solutions with initial pH of 10.5/4 significantly increased the presence of carboxylic acid groups and after Ag attachment the coating had higher antimicrobial activity against both Gram-negative and Gram-positive bacteria compared to the LDPE LbL coated using PEI/PAA polymer solutions with initial pH of 9/6.5. The LDPE LbL coated films using non-modified pH PEI/PAA polymer solutions decreased the water contact-angle indicating an increased hydrophilicity of the film, also increased the tensile strength and roughness of LDPE LbL coated films compared to uncoated LbL samples. The LDPE LbL coated films attached with Ag(+) were UV/ozone treated for 20 min to oxidise Ag(+) to Ag(0). The presence of Ag(0) (Ag nanoparticles (NPs)) on the LDPE LbL coated films was confirmed by XRD, UV-vis spectrophotometer and colour changes. The overall results demonstrated that the LbL technique has the potential to be used as a coating method containing antimicrobial Ag NPs and that the manufactured films could potentially be applied as antimicrobial packaging.
Methicillin-susceptible Staphylococcus aureus (MSSA) is an important nosocomial pathogen worldwide. This study aims to investigate the in vitro biofilm-forming ability of clinical MSSA isolated from various sources in the main public tertiary referral hospital in Terengganu, Malaysia and to detect the presence of biofilm-associated and regulatory genes among these isolates. A total of 104 MSSA isolates [pus (n = 75), blood (n = 24), respiratory secretions (n = 2), eye (n = 2), and urine (n = 1)] were investigated for slime production and biofilm formation using Congo red agar and crystal violet microtitre plate, respectively. Fifteen MSSA isolates with varying degrees of biofilm formation were selected for validation via a real-time cell analyser. All isolates were screened for microbial surface components recognising adhesive matrix molecules (MSCRAMM) and accessory gene regulator (agr) using polymerase chain reaction assay. A total of 76.0% (79/104) isolates produced slime layer, while all isolates developed biofilm as follows: 52.8% (55/104) strong biofilm producers, 40.4% (42/104) intermediate biofilm producers, and 6.7% (7/104) weak biofilm producers. A total of 98.1% (102/104) isolates carried at least one of the screened MSCRAMM gene(s) with the eno gene detected at the highest rate (87.5%, 91/104), while the sasG gene was significantly associated with strong biofilm production (p = 0.015). Three agr groups, 1, 2, and 3, were detected among the MSSA isolates with a predominance of agr-3 (32.7%, 34/104). In conclusion, biofilm formation varied greatly among clinical MSSA isolates, and the presence of sasG gene and agr-1 may play important role in initiating MSSA infections via biofilm formation.
Enterotoxin production by strains of Staphylococcus aureus isolated from foods unconnected with outbreaks offood poisoning was investigated. Twenty-three percent of 217 strains examined produced enterotoxins A, B, C, D or E. Enterotoxin C was found to occur most frequently. Enterotoxin A was not detected alone from any of the strains examined, but occurred together with other enterotoxins. The overall number of strains isolated from raw foods which produced one or more enterotoxins was higher than that for cooked foods. Antibiotic sensitivities were unrelated to enterotoxin production and no correlation could be found between methicillin resistance and enterotoxigenicity.
An experiment was designed to study the in vivo effect of Pasteurella haemolytica A2 infection on the phagocytosis activity of caprine broncho-alveolar macrophages and the extent of pneumonic lesions. Twelve healthy local Kacang goats, about 7 months of age, were divided into two groups of six. Goats in group 1 were inoculated intratracheally with 4 ml inoculum containing 2.8 x 10(9) colony-forming units (CFU)/ml of Staphylococcus aureus. Goats in group 2 were inoculated intratracheally with 4 ml of inoculum containing 9.5 x 10(8) CFU/ml of Pasteurella haemolytica A2 isolated earlier from pneumonic lungs of goat. At intervals of 3 and 7 days post-challenge five goats from each group were killed and the lungs were washed with sterile phosphate-buffered saline. Smears were prepared from the lung washing fluid and the number of macrophages with phagocytic activity was determined. At day 3 post-infection, goats of both groups showed a similar pattern of pneumonic lesion. The lung washing fluid of goats in group 2 was found to contain numerous neutrophils and macrophages. Goats in group 2 showed significantly (P < 0.05) higher extent of lung lesions than group 1. Similarly, the average extent of lung lesions was significantly (P < 0.05) more severe in group 2 at day 7 post-infection. The lung washing fluid contained mostly macrophages. The phagocytic activity following S. aureus infection was more efficient and significantly (P < 0.01) higher compared with infection by P. haemolytica A2. There were weak correlations between the extent of pneumonic lesion and the phagocytic activity. Thus, goats with poor phagocytic activity were likely to develop more extensive lung lesions.
The shortage of new antibiotics makes infections caused by gram-negative (G-) bacteria a significant clinical problem. The key enzymes involved in folate biosynthesis represent important targets for drug discovery, and new antifolates with novel mechanisms are urgently needed. By targeting to dihydrofolate reductase (DHFR), a series of 1,3-diamino-7H-pyrrol[3,2-f]quinazoline (PQZ) compounds were designed, and exhibited potent antibacterial activities in vitro, especially against multi-drug resistant G- strains. Multiple experiments indicated that PQZ compounds contain a different molecular mechanism against the typical DHFR inhibitor, trimethoprim (TMP), and the thymidylate synthase (TS) was identified as another potential but a relatively weak target. A significant synergism between the representative compound, OYYF-175, and sulfamethoxazole (SMZ) was observed with a strong cumulative and significantly bactericidal effect at extremely low concentrations (2 μg/mL for SMZ and 0.03 pg/mL for OYYF-175), which could be resulted from the simultaneous inhibition of dihydropteroate synthase (DHPS), DHFR and TS. PQZ compounds exhibited therapeutic effects in a mouse model of intraperitoneal infections caused by Escherichia coli (E. coli). The co-crystal structure of OYYF-175-DHFR was solved and the detailed interactions were provided. The inhibitors reported represent innovative chemical structures with novel molecular mechanism of action, which will benefit the generation of new, efficacious bactericidal compounds.
The emerging polymer nanocomposites have received industrial interests in diverse fields because of their added value in metal oxide-based nanocomposites, such as titanium (TiO2) and zinc oxide (ZnO). Linear low-density polyethylene (LLDPE)-based polymer has recently generated a huge market in the healthcare industry. TiO2 and ZnO are well known for their instant photocatalytic killing of hospital-acquired infections, especially multidrug-resistant (MDR) pathogens. This study investigated the actions of LLDPE/TiO2/ZnO (1:3) nanocomposites in different weight% against two representative MDR pathogens, namely, methicillin-resistant Staphylococcus aureus (MRSA) and Klebsiella pneumonia (K.pneumoniae). Antibacterial activities were quantified according to international standard guidelines of CLSI MO2-A11 (static condition) and ASTM E-2149 (dynamic condition). Preliminary observation via a scanning electron microscope revealed that LLDPE matrix with TiO2/ZnO nanocomposites changed the bacterial morphology and reduced the bacterial adherence and biofilm formation. Furthermore, a high ZnO weight ratio killed both types of pathogens. The bactericidal potential of the nanocomposite is highlighted by the enhancements in photocatalytic activity, zinc ion release and reactive species, and bacteriostatic/bactericidal activity against bacterial growth. This study provides new insights into the MDR-bactericidal potential of LLDPE with TiO2/ZnO nanocomposites for targeted healthcare applications.
The objective of this study is to determine the therapeutic efficacy of allicin against Candida albicans (C. albicans) and Staphylococcus aureus (S. aureus), the common etiological agents for denture stomatitis (DS). The minimum inhibitory concentration (MICs), minimum bactericidal concentrations (MBCs) and minimum fungicidal concentration (MFCs) of allicin were determined by the broth microdilution method followed by checkerboard microdilution method for a synergistic interaction between allicin + nystatin and allicin + CHX. The potential of allicin to eradicate C. albicans and S. aureus biofilms was assessed by treating biofilm formed on self- polymerized acrylic resin with allicin at a sub-MIC concentration for 5 min. The commercial denture cleanser (brand X) was used as a positive control. A Kruskal-Wallis test followed by the post-hoc Mann-Whitney U test was applied (SPSS 20.0), and the level of significance was set at P
Increasing risk of antibiotic resistance of pathogenic bacteria has led to the exploration of alternative antibiotics derived from leaves of medicinal plants such as the fig (Ficus carica L.). The aim of this study was to determine the antimicrobial activity of the methanolic extract of fig leaves grown under Malaysian tropical environment against pathogenic bacteria linked to antibiotic resistance namely the Staphylococcus aureus and Escherechia coli. The methanolic extraction was performed by using soxhlet apparatus. The disc diffusion method was used to measure inhibition zone diameter on the Mueller-Hinton agar plate. Staphylococcus aureus displayed the highest diameter of inhibition zone against the extract at concentration of 900 mg/ml whilst Escherechia coli displayed the highest diameter of inhibition zone against both the 100% crude extract and 700 mg/ml, respectively. This study therefore highlighted the potential of developing alternative antibiotics derived from the methalonic extract of locally grown fig plant.
Introduction:Staphylococcus aureus is a Gram-positive staphylococci that form biofilms. Bacteria that dwell in bio-films tend to be highly resistant towards the action of antibiotics. S. aureus is a main cause of infections in the oral cavity such as angular cheilitis, endodontic infections, osteomyelitis of the jaw, parotitis and oral mucositis. Previous studies reported that S. aureus also spread to the other parts of the body through the circulatory system, which may lead to chronic infections. Hence the search for new antibacterial agents remains high and needs urgent attention to treat this problem. Plants offer a rich source of antimicrobial agents and bioactive compounds. In this study, aque-ous oil palm leaf extracts (OPLE) has been used as an alternative antibacterial agent against oral infections mainly caused by Staphylococcus aureus. Many studies report the potential use of oil palm leaf extracts in treating bacterial infections such as Escherichia coli, Salmonella sp., Staphylococcus aureus (isolated from other part of the body), Pseudomonas aeruginosa and Bacillus sp. Although previous studies have documented the antimicrobial properties of oil palm leaf extracts, to date no study has been reported on the effect of oil palm leaf extract on oral microbes. Methods: The agar diffusion method, minimum inhibitory concentration (MIC) and minimal bactericidal concen-tration (MBC) assay were conducted in order to observe the antibacterial activity of aqueous oil palm leaf extract. The crystal violet assay was used to determine the anti-biofilm activity of the extracts. Chlorhexidine and deionised distilled water were used as the positive and negative control respectively. For agar diffusion method, the diameter of inhibition zone was measured. Results: The inhibition zone of the tested bacteria was observed between 0-20mm. The MIC and MBC assay were used to know the lowest concentrations of the extract that inhibit the growth and killed the tested bacteria respectively. The MIC and MBC values for the tested bacteria were observed between 0-7.813mg/mL. While for anti-biofilm assays, OPLE aqueous extract acts as a potent anti-biofilm agent with dual actions, pre-venting and eradicating the biofilm of the tested bacteria. Conclusion: In conclusion, the tested plant extracts could serve as alternative natural antibacterial and anti-biofilm agent against oral infections.