Methicillin-resistant Staphylococcus aureus (MRSA) is known to cause nosocomial infections and is now becoming an emerging problem in veterinary medicine. The objective of the study was to determine the presence of MRSA in 100 cats and dogs sampled between November 2007 and April 2008 at the University Veterinary Hospital, Universiti Putra Malaysia. MRSA was detected in 8% of pets sampled. Ten percent (5/50) and 6% (3/50) of the isolates were from dogs and cats, respectively. All MRSA isolates possessed the mecA gene and were found to be resistant to at least three antimicrobials with a minimum of Oxacillin MIC of 8 μg/mL. One isolate (CT04) had an extremely high MIC of >256 μg/mL. The MLST type ST59 found in this study have been reported earlier from Singapore and other countries as a strain from animal and community-associated MRSA respectively. Pulsed-field gel electrophoresis revealed five pulsotypes. Two isolates from cats (CT27 and CT33) and three isolates from dogs (DG16, DG20, and DG49) were respectively assigned to pulsotypes B and D. The study suggests that cats and dogs in Malaysia are potential reservoirs for MRSA.
Mesosomes of Staphylococcus aureus ATCC 25923 treated with antibiotics were examined morphologically under the electron microscope. The Transmission Electron Microscope Rapid Method was used to eliminate the artifacts due to sample processing. Mesosomes were seen in all the antibiotic treated bacteria and not in the control group. The main factor that contributes to the formation of mesosomes in the bacteria was the mode of action of the antibiotics. The continuous cytoplasmic membrane with infolding (mesosomes) as in the S. aureus ATCC 25923 is therefore confirmed as a definite pattern of membrane organization in gram positive bacteria assaulted by amikacin, gentamicin, ciprofloxacin, vancomycin and oxacillin antibiotics. Our preliminary results show oxacillin and vancomycin treated bacteria seemed to have deeper and more mesosomes than those treated with amikacin, gentamicin and ciprofloxacin. Further research is needed to ascertain whether the deep invagination and the number of mesosomes formed is associated with the types of antibiotic used.
Medicinal plants have many traditional claims including the treatment of ailments of infectious origin. In the evaluation of traditional claims, scientific research is important. The objective of the study was to determine the presence of antibacterial activity in the crude extracts of some of the commonly used medicinal plants in Malaysia, Andrographis paniculata, Vitex negundo, Morinda citrifolia, Piper sarmentosum, and Centella asiatica. In this preliminary investigation, the leaves were used and the crude extracts were subjected to screening against five strains of bacteria species, Methicillin Resistant Staphylococcus aureus (MRSA), Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa and Escherichia coli, using standard protocol of Disc Diffusion Method (DDM). The antibacterial activities were assessed by the presence or absence of inhibition zones and MIC values. M. citrifolia, P. sarmentosum and C. asiatica methanol extract and A. paniculata (water extract) have potential antibacterial activities to both gram positive S. aureus and Methicillin Resistant S. aureus (MRSA). None of the five plant extracts tested showed antibacterial activities to gram negative E. coli and K. pneumoniae, except for A. paniculata and P. sarmentosum which showed activity towards P. aeruginosa. A. paniculata being the most potent at MIC of 2 g/disc. This finding forms a basis for further studies on screening of local medicinal plant extracts for antibacteria properties.
Resistant and enterotoxigenic Staphylococcus aureus strains are considered to be one of the major causes of foodborne diseases due to the consumption of sweet. The present research was done to study the distribution of enterotoxin types, enterotoxigenic genes and antibiotic resistance pattern of S. aureus strains isolated from traditional sweet samples. Eight-hundred and fifteen sweet samples were cultured and S. aureus strains were identified. Antibiotic resistance, enterotoxigenicity and enterotoxigenic gene profile were studied using disk diffusion, Enzyme Link Immunosorbent Assay and PCR, respectively. One-hundred and seven out of 815 (13.12%) sweet samples were positive for S. aureus. Prevalence of S. aureus in dried and semi-dried sweet samples were 15.08% and 11.13%, respectively (P <0.05). Forty-six out of 107 S. aureus strains (42.99%) were determined as enterotoxigenic. A (41.30%) and C (17.39%) were the most commonly detected enterotoxin types. Sea (20.56%), sec (14.95%) and seb (11.21%) were the most commonly detected enterotoxigenic genes. There were no positive sample for the sej enterotoxin gene. S. aureus strains harbored the highest prevalence of resistance against penicillin (88.78%), tetracycline (83.17%), ceftaroline (75.70%) and doxycycline (71.02%). Simultaneous presence of enterotoxins and enterotoxigenic genes in multi-drug resistant S. aureus strains indicates important public health issue regarding the consumption of contaminated traditional sweet samples.
The bioactivity of R. nasutus leaf extracts was assessed on Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Streptococcus pyogenes, Vibrio parahaemolyticus, Enterobacter aerogenes, Proteus mirabilis, and Klebsiella pneumoniae. Crude chloroform, petroleum ether, ethyl acetate, ethanol and methanol extracts were screened by disc diffusion method. Promising crude extract was further subjected to the column fractionation followed by the screening of the antibacterial activity of individual fractions. Biologically active pure fraction was subjected to the advanced analytical studies like HPLC, LC-MS, IR and NMR for characterisation of the bioactive compound. Ethanolic extract exhibited the maximum antibacterial activity against Klebsiella pneumoniae with the maximum of 35±0.42 mm zone of inhibition. The biologically potent column fraction from ethanol extract with 40±0.42 mm zone of inhibition upon subject to the HPLC, LC-MS, IR and NMR revealed that the active compound is rhinacanthin-C, a naphthoquinone.
Hand hygiene is the topmost crucial procedure to prevent hospital-acquired infections. Choosing an effective hand disinfectant is necessary in enforcing good hand hygiene practice especially in hospital settings. The aim of the study was to investigate the efficacy of Aaride AGT-1 as a hand disinfectant for the inhibition of pathogenic microorganisms' transmission among both patients and personnel in the health care system compared to other commercially available disinfectants. In the present study, a new hand disinfectant Aaride AGT-1 was tested against several bacterial and viral pathogens to evaluate its antimicrobial activity profile. The results revealed that Aaride AGT-1 displayed the highest antibacterial activity against five pathogenic bacteria including MRSA when compared to other commercially available hand sanitizers. Aaride AGT-1 showed the lowest percentage needed to inhibit the growth of bacterial pathogens. In addition, results obtained from time killing assay revealed that Aaride AGT-1 demonstrated the best killing kinetics, by eradicating the bacterial cells rapidly within 0.5 min with 6 log reduction (>99.99% killing). Also, Aaride AGT1 was able to reduce 100% plaque formed by three viruses namely HSV-1, HSV-2 and EV-71. In conclusion, Aaride AGT-1 is capable of killing wide-spectrum of pathogens including bacteria and viruses compared to other common disinfectants used in hospital settings. Aaride AGT-1's ability to kill both bacteria and viruses contributes as valuable addition to the hand disinfection portfolio.
The emergence of methicillin-resistant Staphylococcus aureus (MRSA) infection is one of the greatest threats to both animal and human health. Our investigation was aimed to identify and differentiate between MRSA and methicillin-sensitive Staphylococcus aureus (MSSA) recovered from mastitic milk using MALDI-TOF mass spectrometry compared with phenotypic methods and studying their susceptibility to various antibiotics. Four hundred milk samples from mastitic animals (cows, sheep, goats, and dromedary camels) were investigated. Phenotypic identification of S. aureus was made through MASTASAPH Latex test, STAPH ID 32, and Vitek 2 system. The proteomic characterization of S. aureus was done by MBT. The Kirby Bauer method was accomplished to detect the resistance of S. aureus strains to antibiotics. The results of the MASTASAPH Latex test, revealed that 54 (46%) were recognized as S. aureus. All S. aureus isolates were identified by MBT with a score of more or equal 2.00. Several peaks were identified in the mass of 4590 Da, 4863 Da, and 4938 Da for MSSA and in the mass of 2636 Da and 3009 Da for MRSA. The MSP dendrogram demonstrated that the S. aureus isolates were classified into one group with a distance level of less or equal 400. The percentage of S. aureus resistance against carbenicillin, erythromycin and kanamycin was 94.4%, 38.88%, and 33.33%, respectively. In conclusion, S. aureus bacteria are among the key triggers for mastitis in Saudi Arabia. MBT is reported to be not only the rapid tool to identify S. aureus but also able to discriminate MRSA from MSSA.
Bacterial mediated Silver nanoparticles is considered as an emerging Ecofriendly approach to eradicate human pathogens. This paper aims to provide the biological approach for the synthesis of silver nanoparticles from indigenously isolated bacteria. This study will be beneficial to control the nosocomial infections triggered by MRSA (Methicillin-resistant Staphylococcus aureus). The current study is the extracellular synthesis of silver nanoparticles by using the cell free filtrate of bacterial strains isolated from the soil. The optimization study was also carried out to obtain the maximum production of silver nanoparticles. Nanoparticles were confirmed and characterized by UV-Vis spectroscopy and Transmission Electron Microscopy (TEM) having the plasmon resonance peak between 420-450nm with 10-60nm in size range and most were spherical in shape. Synthesized silver nanoparticles showed a potential antibacterial activity against MRSA (Methicillin Resistant Staphylococcus aureus) in-vitro study. This is the green approach for the production of AgNPs, as there was no previous work done on the synthesis of silver nanoparticles by bacteria in this region of Southern Punjab, Pakistan and these nanoparticles can be used to treat nosocomial infection. These silver nanoparticles can be used in effective disease management as antimicrobial agent.
This study aimed to determine the in vivo effectiveness of the ethanolic extract of Piper betle L. leaves against Staphylococcus aureus-infected wounds in mice and its antimicrobial properties on clinical isolates of multiple drug-resistant bacterial pathogens. Twenty mice were divided into four groups. Wounds were created in all mice under anesthesia by excision from the dorsal skin down to the subcutaneous fat and inoculating with S. aureus. After 24 h, the wound of each mouse was treated once daily by application of the respective cream. Group I was treated with mupirocin antibacterial cream; Group II received a cream base containing no active ingredient; Groups III and IV were treated with 2.5% and 5.0% concentrations of P. betle cream, respectively. Further, an in vitro study was performed by adding undiluted, 1:50 and 1:100 dilutions of the four studied creams in normal saline containing 1.5 × 108 CFU/mL of the following bacteria: antimicrobial-susceptible S. aureus, Escherichia coli, Pseudomonas aeruginosa, methicillin-resistant S. aureus, extended-spectrum β-lactamase-producing Escherichia coli, vancomycin-resistant Enterococcus, metallo-βlactamase-producing P. aeruginosa and carbapenem-resistant Klebsiella pneumoniae. The mice in Groups III and IV had significantly faster wound contraction and significantly shorter reepithelialization time than Group II (p < 0.05), which were not significantly different from Group I (p > 0.05). P. betle creams inhibited all studied bacterial strains at full concentration and at a dilution of 1:50. The inhibitory effect was more significant than Groups I and II (p < 0.05), except on S. aureus. Specifically, S. aureus inhibition was not significantly different for Groups III and IV (p > 0.05) when compared with Group I. Cream formulations derived from P. betle ethanolic extract have great potential as antimicrobial agents for the treatment of wound infection. Further clinical tests are recommended to determine the safety and efficacy of these formulations in other mammalian species.
The present study was conducted to investigate the antimicrobial potential of essential oils of Curcuma longa and Syzygium aromaticum against multidrug-resistant pathogenic bacteria. Four identified bacterial isolates including Methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, Klebsiella pneumoniae, and Acinetobacter baumannii were selected and their antibiotic sensitivity was checked by disc diffusion assay. C. longa and S. aromaticum were subjected to steam distillation to obtain their essential oils. The crude essential oils were fractioned by employing column chromatography. Crude essential oils and their fractions were evaluated for their antibacterial activity by agar well diffusion assay and minimum inhibitory concentrations were calculated. All the selected bacterial isolates showed resistance to three or more than three antibiotic groups and were declared as multidrugresistant (MDRs). Crude essential oils of C. longa and S. aromaticum exhibited antimicrobial activity against all selected isolates but S. aromaticum activity was better than the C. longa with a maximum 19.3±1.50 mm zone of inhibition against A. baumannii at 1.04 µL/mL MIC. GC/MS analysis revealed the abundance of components including eugenol, eugenyl acetate, b- caryophyllene, and a- Humulene in both crude oil and fractions of S. aromaticum. While the main components of C. longa essential oil were Ar-tumerone, a-tumerone, b- Tumerone, I-Phellandrene, a-zingibirene, b- sesquiphellandrene, and p- Cymene. This study highlights that plant-based essential oils could be a promising alternative to antibiotics for which pathogens have developed resistance. C. longa and S. aromaticum carry compounds that have antimicrobial potential against multiple drug-resistant bacteria including MRSA. E. coli, K. pneumoniae and A. baumannii.
Mastitis is the inflammation of the mammary gland due to microbial infiltration causing a reduced mammary function. This study aims at developing a vaccine using Malaysian local isolate of Staphylococcus aureus and evaluating serum amyloid A, Interleukin-10, IgM and IgG responses periodically. Four bacterin concentrations (106, 107, 108 and 109 cfu/ml of the local isolate of S. aureus) were adjuvanted with aluminium potassium sulphate. Thirty cows grouped into 4 treatment groups (G-) were vaccinated (2 ml) intramuscularly, with a fifth G-A as control. The mean concentration (MC) of serum amyloid A (SAA) was significantly different (sig-d) (p ˂ 0.05) in G-D at 0 h post vaccination (PV), 3 h PV, 24 h PV, weeks 1, 2, 3 and 4 PV (6-, 15-, 5-, 12-, 11-, 4- and 11-fold increased (FI) respectively). The MC of serum amyloid A was also sig-d in G-E at 0 h PV, weeks 1, 2 and 4 PV (3, 8, 5 and 8 FI respectively). The MC of IL-10 was sig-d in G-D and C at 3 h PV and week 2 PV (5 and 2 FI respectively). The IgM MC was sig-d in G-B and C at 3 h PV (5 and 6 FI respectively), at 24 h PV (5 and 9 FI respectively), at week 3 PV(2 and 2 FI respectively) and week 4 PV (3 and 4 FI respectively). The MC of IgG was sig-d in G-E at 0 h, 3 h and week 3 PV(5, 6 and 2 FI respectively) and in G-D at weeks 1-4 (3, 3, 3 and 5 FI respectively). In conclusion, elevated levels of SAA, IgG and IL-10 in G-D(108) informed our choice of best dosage which can be used to evoke immunity in cows.
The epidermal mucus of fish contains antimicrobial agents that act as biological defence against disease. This study aims to identify antibacterial activity and protein concentration of epidermal mucus of Barbodes everetti, a Bornean endemic freshwater fish. The epidermal mucus was extracted with 3% acetic acid, 0.85% sodium chloride and crude solvents. The mucus activity against eight strains of human pathogenic bacteria, including Bacillus cereus ATCC 33019, Escherichia coli O157:H7, Listeria monocytogenes ATCC 7644, Pseudomonas aeruginosa ATCC 27853, Salmonella braenderup ATCC BAA 664, Salmonella typhimurium, Staphylococcus aureus ATCC 25933, and Vibrio cholerae, were tested. The acetic acid mucus extract of B. everetti was able to inhibit five strains of bacteria and show no activity toward E. coli O157:H7, B. cereus ATCC 33019 and L. monocytogenes ATCC 7644. Moreover, the highest protein concentration was quantified in crude extract, followed by aqueous and acetic acid extracts. This study provides a preliminary knowledge on the activity of epidermal mucus of B. everetti towards five out of the eight human pathogens tested, therefore it may contain potential sources of novel antibacterial components which could be further extracted for the production of natural antibiotics towards human-related pathogenic bacteria.
The prevalence of resistance to rifampicin and fusidic acid among Malaysian strains of methicillin-resistant Staphylococcus aureus (MRSA) is increasing. This study aimed to determine the mechanisms of rifampicin and fusidic acid resistance and the genetic diversity of MRSA strains from a Malaysian tertiary hospital.
Abstract. Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) is a pathogen recognized to be distinct in both phenotype and genotype from hospital-acquired MRSA. We have identified CA-MRSA cases in Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia, including their antibiotic susceptibility patterns and genotypic characteristics. Cases were identified during January to December 2009 from routine clinical specimens, where culture and antibiotic susceptibility results yielded pauci-resistant MRSA isolates suspected as being CA-MRSA. The patients' clinical data were collected and their specimens were sent for molecular confirmation and analysis. Five cases of CA-MRSA were identified, which had a multi-sensitive pattern on antibiotic susceptibility tests and were resistant to only penicillin and oxacillin. All cases were skin and soft-tissue infections, including diabetic foot with gangrene, infected scalp hematoma, philtrum abscess in a healthcare worker, thrombophlebitis complicated with abscess and infected bedsore. All five cases were confirmed MRSA by detection of mecA. SCCmec typing (ccr and mec complex) revealed SCCmec type IV for all cases except the infected bedsore case. Panton-Valentine leukocidin gene was positive in all isolates. As clinical features among methicillin-sensitive Staphylococcus aureus, CA-MRSA and "nosocomial CA-MRSA" are indistinct, early recognition is necessary in order to initiate appropriate antibiotics and infection control measures. Continual surveillance of pauci-resistant MRSA and molecular analysis are necessary in order to identify emerging strains as well as their epidemiology and transmission, both in the community and in healthcare setting.
Methicillin-resistant Staphylococcus aureus (MRSA), an established nosocomial and emerging community pathogen associated with many fatalities due to its hyper-virulence and multiple drug resistant properties, is on the continuous rise. To update the current status on the susceptibility of local MRSA isolates to various classes of antibiotics and to identify the most potent antibiotics, thirty-two clinical isolates comprised of hospital acquired (HA) and community acquired (CA) infections were investigated by disk diffusion test. Of the 32 MRSA isolates, 14 (43.75%) and 18 (56.25%) were community and hospital acquired MRSA, respectively. All isolates were multiple drug resistant to more than 3 classes of antibiotics despite the source or specimen from which it was isolated. The oxacillin MICs for all isolates ranged from 2 to > or = 256 microg/ml. Twenty-five of 26 erythromycin-resistant MRSA isolates exhibited an inducible MLS(B) resistance phenotype while one showed an MS phenotype. More than half the isolates (68.75%) were resistant to at least one of the six aminoglycosides tested, with netilmicin as the most susceptible. The most effective antistaphylococcal agents were linezolid, vancomycin, teicoplanin and quinupristin/dalfopristin exhibited 100% susceptibility. Since MRSA is under continuous pressure of acquiring multiple drug resistance, it is imperative to focus routine surveillance on HA and CA-MRSA strains to monitor and limit the spread of this organism.
Neonatal septic arthritis has always been considered as separate from its counterpart in older children. The condition is uncommon but serious. Affected neonates usually survive, but with permanent skeletal deformities. Ten cases of neonatal septic arthritis were diagnosed between January 1989 and December 1993 in the neonatal intensive care units of two referral hospitals in the state of Kelantan, Malaysia. All except one neonate was born prematurely. The mean age of presentation was 15.6 days. Joint swelling (10/10), increased warmth (7/10) and erythema of the overlying skin (7/10) were the common presenting signs. Vague constitutional symptoms preceded the definitive signs of septic arthritis in all cases. The total white cell counts were raised with shift to the left. The knee (60%) was not commonly affected, followed by the hip (13%) and ankle (13%). Three neonates had multiple joint involvement. Coexistence of arthritis with osteomyelitis was observed in seven neonates. The commonest organism isolated was methicillin resistant Staphylococcus aureus (9/10). Needle aspiration was performed in nine neonates and one had incision with drainage. Follow up data was available for five neonates and two of these had skeletal morbidity. Early diagnosis by frequent examination of the joints, prompt treatment and control of nosocomial infection are important for management.
Enterotoxin production by strains of Staphylococcus aureus isolated from human, food and animal sources was investigated. Of the 130 isolates studied, 27 (20.8%) were found to be enterotoxigenic. The most common enterotoxin detected from human sources was enterotoxin C whereas enterotoxin B occurred more frequently in staphylococcal strains of food origin. The 2 enterotoxigenic strains, from animals isolated from a dog and a goat, produced enterotoxins A and C, respectively. Enterotoxin E was not detected alone from any of the enterotoxigenic strains studied, but occurred together with other enterotoxins. The need to detect enterotoxin in staphylococcal strains and in suspected foods for the confirmation of staphylococcal food poisoning is discussed.
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
We report a case study on a patient who presented with low back and thigh pain of one month duration. He was eventually diagnosed with left thigh pyomyositis. Tissue from thigh grew Staphylococcus aureus. With commencement of antibiotics and surgical drainage, patient made recovery despite prolonged hospital stay. The underlying mechanism of the extensive abscess accompanied by lack of systemic symptoms; is related to relative immunocompromised state of having underlying diabetes mellitus.