Sternal wound infection is an infrequent yet potentially devastating complication following sternotomy. Among the standard practices used as preventive measures are the use of prophylactic antibiotics and povidone-iodine as an irrigation agent. A new antiseptic agent, Dermacyn super-oxidized water (Oculus Innovative Sciences), has recently been used as a wound-irrigation agent before the closure of sternotomy wounds.
Acinetobacter spp is a known nosocomial pathogen causing a wide range of clinical diseases such as pneumonia, wound infection and bloodstream infections (BSI). The clinical outcomes of acinetobacter BSI were determined by a 1:1 case control study involving 58 confirmed cases of acinetobacter BSI who were compared to other gram-negative infections. The crude mortality of acinetobacter BSI was 47.2%, which was significantly greater than other gram-negative BSI (OR 1.89, 95% CI 1.10-3.24) but there were no significant differences in attributed mortality between the two groups. We found that patients treated in intensive care units (ICU), who had longer ICU stays, who presented with shock or coagulopathy, had prior exposure to carbapenems, had mechanical ventilation, were on a ventilator for longer periods, had a nasogastric tube, had an arterial catheter or had parenteral nutrition at a significantly greater risk of mortality due to acinetobacter BSI. Patients presenting with septic shock (OR 17.95, 95% CI 3.36-95.84) or having a central venous catheter (OR 12.48, 95% CI 1.09-142.68) were independently at higher risk for mortality. Appropriateness of therapy reduced the mortality attributes of acinetobacter BSI (OR 0.197, 95% CI 0.040-0.967) but did not significantly reduce crude mortality in acinetobacter BSI patients. This study shows the importance of preventing acinetobacter BSI and the appropriate use of antimicrobial agents to reduce mortality.
Hospital-acquired vancomycin-resistant enterococci (VRE) were first reported in the late 1980s and have since been an increasing problem worldwide. Kuala Lumpur Hospital thus far, to the best of our knowledge has been spared from this pathogen. We describe the first confirmed case of Enterococcus faecium exhibiting the van A phenotype in our hospital, in a patient with chronic renal failure who was successfully treated with linezolid. The microbiology laboratory plays an important role in the identification and detection of VRE.
We have previously reported the phenotypic characterization of methicillin-resistant Staphylococcus aureus (MRSA) clinical strains isolated in Malaya University Hospital in the period 1987 to 1989 using antibiogram, coagulase typing, plasmid profiles, and phage typing. Here, we report the analysis of the same strains with three genotyping methods; ribotyping, pulsed-field gel electrophoresis (PFGE) typing, and IS431 typing (a restriction enzyme fragment length polymorphism analysis using an IS431 probe). Ribotyping could discriminate 46 clinical MRSA strains into 5 ribotypes, PFGE typing into 22 types, and IS431 typing into 15 types. Since the differences of the three genotyping patterns from strain to strain were quite independent from one another, the combined use of the three genotyping methods could discriminate 46 strains into 39 genotypes. Thus, the powerful discriminatory ability of the combination was demonstrated.
The pattern of antibiotic resistance amongst gram-negative bacteria (GNB) in paediatric units, which have heavy empirical usage of broad-spectrum antibiotics, was studied prospectively over a 6-month period. A total of 200 consecutive, non-duplicate gram-negative isolates were obtained from 109 patients admitted to intensive care and oncology units in two hospitals. The commonest isolates were Klebsiella spp (36.5 per cent) and Pseudomonas (20.0 per cent). The isolates showed lower susceptibility rates to the third-generation cephalosporins (47-62 per cent) compared with cefepime (91 per cent), imipenem (90 per cent) and ciprofloxacin (99 per cent). Fifty-four (52.8 per cent) Klebsiella and Escherichia coli isolates were determined to be extended-spectrum beta-lactamase (ESBL) producing strains. Antibiotics found to be effective against ESBL-producers were imipenem and ciprofloxacin. The high resistance rate amongst GNB to third-generation cephalosporins is a likely consequence of heavy empirical usage of this group of antibiotics. The carbapenems and quinolones remain useful agents in the management of patients admitted to these units.
Pantoea infections are uncommon in humans. Most reports have involved adults or children after thorn injuries. There are only a few reports of systemic infections with Pantoea. This is the first report of the clinical picture of systemic Pantoea spp. infection in neonates as observed during an outbreak in a neonatal intensive care unit caused by infected parenteral nutrition solutions. Even though detected early, the infections had a fulminant course, causing septicemic shock and respiratory failure. Pulmonary disease was prominent and presented mainly as pulmonary hemorrhage and adult respiratory distress syndrome. The organism was sensitive to most antibiotics used in neonatal intensive care units, but the clinical response to antibiotic therapy was poor. The fatality rate was very high: 7 out of 8 infected infants succumbed to the infection (87.5%).
Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most common nosocomial pathogens, causing mild to severe infections. This study aimed to determine the genotypic and phenotypic characteristics of clinical MRSA isolates collected from a teaching hospital from 2014 - 2015. These isolates were genotyped by multilocus sequence typing, staphylococcal cassette chromosomal mec (SCCmec) typing, virulence genes detection, and pulsed-field gel electrophoresis; they were phenotyped based on their antibiotics susceptibility profiles. The most prevalent sequence type was ST22. ST3547 was identified from a blood isolate from 2015. Three SCCmec types (III in 26.26%, IV in 70.71%, and V in 3.03% isolates) were detected. Agr type I, II, and III were also detected among the isolates. The most prevalent virulence genes found were hemolysin (100%) and intracellular adhesion (91.9%). At least one staphylococcal enterotoxin was detected in 83 (83.8%) isolates. All the isolates were susceptible to vancomycin (minimum inhibitory concentration ≤ 2 μg/mL). Statistical analysis revealed a significant increase in hypertension (p = 0.035), dyslipidemia and obesity (p = 0.046), and previous exposure to any quinolone (p = 0.010) cases over the two-year period. The emergence and circulation of community-associated MRSA variants were observed in our hospital.
The annual prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in Malaysia has been estimated to be 30 % to 40 % of all S. aureus infections. Nevertheless, data on the antimicrobial resistance and genetic diversity of Malaysian MRSAs remain few. In 2009, we collected 318 MRSA strains from various wards of our teaching hospital located in Kuala Lumpur, the capital city of Malaysia, and performed antimicrobial susceptibility testing on these strains. The strains were then molecularly characterized via staphylococcal cassette chromosome (SCC) mec and virulence gene (cna, sea, seb, sec, sed, see, seg, seh, sei, eta, etb, Panton-Valentine leukocidin and toxic shock syndrome toxin-1) typing; a subset of 49 strains isolated from the intensive care unit was also typed using PFGE. Most strains were found to be resistant to ciprofloxacin (92.5 %), erythromycin (93.4 %) and gentamicin (86.8 %). The majority (72.0 %) of strains were found to harbour SCCmec type III-SCCmercury with the presence of ccrC, and carried the sea+cna gene combination (49.3 %), with cna as the most prevalent virulence gene (94.0 %) detected. We identified four PFGE clusters, with pulsotype C (n=19) as the dominant example in the intensive care unit, where this pulsotype was found to be associated with carriage of SCCmec type III and the sea gene (P=0.05 and P=0.02, respectively). In summary, the dominant MRSA circulating in our hospital in 2009 was a clone that was ciprofloxacin, erythromycin and gentamicin resistant, carried SCCmec type III-SCCmercury with ccrC and also harboured the sea+cna virulence genes. This clone also appears to be the dominant MRSA circulating in major hospitals in Kuala Lumpur.
This study investigated 147 multidrug-resistant Enterobacteriaceae and Pseudomonas aeruginosa isolates from hospitalized patients in Malaysia. Class 1 integrons were the most dominant class identified (45.6%). Three isolates were shown to contain class 2 integrons (2.0%), whilst one isolate harboured both class 1 and 2 integrons. No class 3 integrons were detected in this study. In addition, the sul1 gene was amplified in 35% of isolates and was significantly associated with the presence of integrase genes in an integron structure. RFLP and DNA sequencing analyses revealed the presence of 19 different cassette arrays among the detected integrons. The most common gene cassettes were those encoding resistance towards aminoglycosides (aad) and trimethoprim (dfr). As far as is known, this study is the first to identify integron-carrying cassette arrays such as aadA2-linF, aacC3-cmlA5 and aacA4-catB8-aadA1 in the Malaysian population. Patients' age was demonstrated as a significant risk factor for the acquisition of integrons (P=0.028). Epidemiological typing using PFGE also demonstrated a clonal relationship among isolates carrying identical gene cassettes in Klebsiella pneumoniae and P. aeruginosa but not in Escherichia coli isolates.
Multidrug-resistant (MDR) Acinetobacter baumannii has increasingly emerged as an important nosocomial pathogen. The aim of this study was to determine the resistance profiles and genetic diversity in A. baumannii clinical isolates in a tertiary medical center in Malaysia. The minimum inhibitory concentrations of carbapenems (imipenem and meropenem), cephalosporins (ceftazidime and cefepime), and ciprofloxacin were determined by E-test. PCR and sequencing were carried out for the detection of antibiotic resistance genes and mutations. Clonal relatedness among A. baumannii isolates was determined by REP-PCR. Sequence-based typing of OXA-51 and multilocus sequence typing were performed. One hundred twenty-five of 162 (77.2%) A. baumannii isolates had MDR phenotype. From the 162 A. baumannii isolates, 20 strain types were identified and majority of A. baumannii isolates (66%, n = 107) were classified as strain type 1 and were positive for ISAba1-blaOXA-23and ISAba1-blaADCand had mutations in both gyrA and parC genes at positions, 83 and 80, resulting in serine-to-leucine conversion. REP-PCR analysis showed 129 REP types that generated 31 clones with a 90% similarity cutoff value. OXA-66 variant of the blaOXA-51-likegenes was predominantly detected among our A. baumannii clinical isolates belonging to ST195 (found in six clones: 1, 8, 9, 19, 27, and 30) and ST208 (found in clone 21). The study helps us in understanding the genetic diversity of A. baumannii isolates in our setting and confirms that international clone II is the most widely distributed clone in Universiti Kebangsaan Malaysia Medical Centre, Malaysia.
Microbial resistance to existing antibiotics has led to an increase in the use of medicinal plants that show beneficial effects for various infectious diseases. The study evaluates the susceptibility of multidrug resistant Staphylococcus aureus to Nigella sativa oil. Staphylococcus aureus was isolated from 34 diabetic patient's wounds attending the Renaissance hospital, Nsukka, Southeast Nigeria. The isolates were characterized and identified using standard microbiological techniques. Isolates were cultured and a comparative In vitro antibiotic susceptibility test was carried out using the disk diffusion method. Of the 34 samples collected, 19(56%) showed multidrug resistance to the commonly used antibiotics. Nigella sativa oil was then studied for antibacterial activity against these multidrug resistant isolates of Staphylococcus aureus in varying concentration by well diffusion method. The oil showed pronounced dose dependent antibacterial activity against the isolates. Out of 19 isolates, 8(42%) were sensitive to undiluted oil sample; 4(21%) of these showed sensitivity at 200 mg/ml, 400 mg/ml and 800 mg/ml respectively. Eleven (58%) of the isolates were completely resistant to all the oil concentrations. The present study, reports the isolation of multi-drug resistant S. aureus from diabetic wounds and that more than half of isolates were susceptible to different concentrations N. sativa oil.
Gelatin hydrogels are attractive for wound applications owing to their well-defined structural, physical, and chemical properties as well as good cell adhesion and biocompatibility. This study aimed to develop gelatin hydrogels incorporated with bio-nanosilver functionalized with lactoferrin (Ag-LTF) as a dual-antimicrobial action dressing, to be used in treating infected wounds. The hydrogels were cross-linked using genipin prior to loading with Ag-LTF and characterized for their physical and swelling properties, rheology, polymer and actives interactions, and in vitro release of the actives. The hydrogel's anti-biofilm and antibacterial performances against S. aureus and P. aeruginosa as well as their cytotoxicity effects were assessed in vitro, including primary wound healing gene expression of human dermal fibroblasts (HDFs). The formulated hydrogels showed adequate release of AgNPs and LTF, with promising antimicrobial effects against both bacterial strains. The Ag-LTF-loaded hydrogel did not significantly interfere with the normal cellular functions as no alteration was detected for cell viability, migration rate, and expression of the target genes, suggesting the nontoxicity of Ag-LTF as well as the hydrogels. In conclusion, Ag-LTF-loaded genipin-cross-linked gelatin hydrogel was successfully synthesized as a new approach for fighting biofilms in infected wounds, which may be applied to accelerate healing of chronic wounds.
The humoral immune responses against 46 different staphylococcal antigens in 27 bacteremia patients infected by clonally related methicillin-resistant Staphylococcus aureus (MRSA) strains of a single sequence type (ST) 239 were investigated. A group of non-infected patients (n = 31) hospitalized for different reasons served as controls. All strains were confirmed as ST 239 by S. aureus and mecA-specific PCR, spa, and multi-locus sequence typing (MLST). In each bacteremia patient, a unique pattern of S. aureus antigen-specific immune responses after infection was observed. Antibody levels among bacteremia patients were significantly higher than controls for HlgB (P = 0.001), LukD (P = 0.009), LukF (P = 0.0001), SEA (P = 0.0001), SEB (P = 0.011), SEC (P = 0.010), SEQ (P = 0.049), IsaA (P = 0.043), IsdA (P = 0.038), IsdH (P = 0.01), SdrD (P = 0.001), SdrE (P = 0.046), EsxA (P = 0.0001), and SA0104 (P = 0.0001). On the other hand, the antibody levels were significantly higher among controls for SSL3 (P = 0.009), SSL9 (P = 0.002), and SSL10 (P = 0.007) when the IgG level on the day of infection was compared with that measured on the day of admission. Diversity was observed in the immune response against the antigens. However, a set of antigens (IsaA, IsdA, IsdH, SdrD, and HlgB) triggered a similar type of immune response in different individuals. We suggest that these antigens could be considered when developing a multi-component (passive) vaccine. SEA and/or its specific antibodies seem to play a critical role during ST239 MRSA bacteremia and SEA-targeted therapy may be a strategy to be considered.
Multidrug-resistant (MDR) and extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae associated with nosocomial infections have caused serious problems in antibiotic management with limited therapeutic choices. This study aimed to determine the genotypic and phenotypic characteristics of K. pneumoniae strains isolated from a tertiary hospital in Malaysia. Ninety-seven clinical K. pneumoniae strains were analyzed for antimicrobial susceptibility, all of which were sensitive to amikacin and colistin (except one strain), while 31.9 % and 27.8 % were MDR and ESBL producers, respectively. PCR and DNA sequencing of the amplicons indicated that the majority of MDR strains (26/27) were positive for blaTEM, followed by blaSHV (24/27), blaCTX-M-1 group (23/27), blaCTX-M-9 group (2/27), and mcr-1 (1/27). Thirty-seven strains were hypervirulent and PCR detection of virulence genes showed 38.1 %, 22.7 %, and 16.5 % of the strains were positive for K1, wabG, and uge genes, respectively. Genotyping by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) showed that these strains were genetically diverse and heterogeneous. Sequence types, ST23, ST22, and ST412 were the predominant genotypes. This is the first report of colistin-resistant K. pneumoniae among clinical strains associated with mcr-1 plasmid in Malaysia. The findings in this study have contributed to the effort in combating the increase in antimicrobial resistance by providing better understanding of genotypic characteristics and resistance mechanisms of the organisms.
Lyophilised wafers have been shown to have potential as a modern dressing for mucosal wound healing. The wafer absorbs wound exudates and transforms into a gel, thus providing a moist environment which is essential for wound healing. The objective of this study was to develop a carboxymethyl cellulose wafer containing antimicrobials to promote wound healing and treat wound infection. The pre-formulation studies began with four polymers, sodium carboxymethyl cellulose (NaCMC), methylcellulose (MC), sodium alginate and xanthan gum, but only NaCMC and MC were chosen for further investigation. The wafers were characterised by physical assessments, solvent loss, microscopic examination, swelling and hydration properties, drug content uniformity, drug release and efficacy of antimicrobials. Three of the antimicrobials, neomycin trisulphate salt hydrate, sulphacetamide sodium and silver nitrate, were selected as model drugs. Among the formulations, NaCMC wafer containing neomycin trisulphate exhibited the most desirable wound dressing characteristics (i.e., flexibility, sponginess, uniform wafer texture, high content drug uniformity) with the highest in vitro drug release and the greatest inhibition against both Gram positive and Gram negative bacteria. In conclusion, we successfully developed a NaCMC lyophilised wafer containing antimicrobials, and this formulation has potential for use in mucosal wounds infected with bacteria.
Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) continue to be a problem for clinicians worldwide. The objective of this study was to determine the changes in antibiograms of MRSA and their genotypic characteristics.
Methicillin-resistant Staphylococcus aureus (MRSA) emerged as a nosocomial pathogen in early 1960s, causing Increasing number of outbreaks in 19708, first reported in a teaching hospital in Malaysia in 1972, causing increased mortality, morbidity, and healthcare costs. Aim of this study is to screen out MRSA from various clinical samples and to see their antibiotic susceptibility pattern. From May 2008 to May 2009, 204 S aureus strains were isolated, out of which 114 (55.8%) were MRSA, and rest methicillin-sensitive Staphylococcus aureus (MSSA). Most of the MRSA strains were obtained from pus (45%) followed by urine (20.5%). Frequency of isolating MRSA were maximum in catheter tip (80%) followed by blood (66.7%) and pus (58.7%). MRSA strains were showing 100% sensitivity to vancomycin and Iinezolid, whereas 92.9% to teicoplanin. Therefore it is concluded that antibiotics other than vancomycin can be used as anti-MRSA agents after sensitivity test, as well as irrational and indiscriminate use of antibiotics can be avoided.