Active Antimicrobial Agents Research Articles

Luminescent Di‐ and Polynuclear Organometallic Gold(I)–Metal (Au2, {Au2Ag}n and {Au2Cu}n) Compounds Containing Bidentate Phosphanes as Active Antimicrobial Agents

The reaction of new dinuclear gold(I) organometallic complexes containing mesityl ligands and bridging bidentate phosphanes [Au(2)(mes)(2)(μ-LL)] (LL=dppe: 1,2-bis(diphenylphosphano)ethane 1a, and water-soluble dppy: 1,2-bis(di-3-pyridylphosphano)ethane 1b) with Ag(+) and Cu(+) lead to the formation of a family of heterometallic clusters with mesityl bridging ligands of the general formula [Au(2)M(μ-mes)(2) (μ-LL)][A] (M=Ag, A=ClO(4)(-), LL=dppe 2a, dppy 2b; M=Ag, A=SO(3)CF(3)(-), LL=dppe 3a, dppy 3b; M=Cu, A=PF(6)(-), LL=dppe 4a, dppy 4b). The new compounds were characterized by different spectroscopic techniques and mass spectrometry The crystal structures of [Au(2)(mes)(2)(μ-dppy)] (1b) and [Au(2)Ag(μ-mes)(2)(μ-dppe)][SO(3)CF(3)] (3a) were determined by a single-crystal X-ray diffraction study. 3a in solid state is not a cyclic trinuclear Au(2)Ag derivative but it gives an open polymeric structure instead, with the {Au(2)(μ-dppe)} fragments "linked" by {Ag(μ-mes)(2)} units. The very short distances of 2.7559(6) Å (Au-Ag) and 2.9229(8) Å (Au-Au) are indicative of gold-silver (metallophilic) and aurophilic interactions. A systematic study of their luminescence properties revealed that all compounds are brightly luminescent in solid state, at room temperature (RT) and at 77 K, or in frozen DMSO solutions with lifetimes in the microsecond range and probably due to the self-aggregation of [Au(2)M(μ-mes)(2)(μ-LL)](+) units (M=Ag or Cu; LL=dppe or dppy) into an extended chain structure, through Au-Au and/or Au-M metallophilic interactions, as that observed for 3a. In solid state the heterometallic Au(2)M complexes with dppe (2a-4a) show a shift of emission maxima (from ca. 430 to the range of 520-540 nm) as compared to the parent dinuclear organometallic product 1a while the complexes with dppy (2b-4b) display a more moderate shift (505 for 1b to a max of 563 nm for 4b). More importantly, compound [Au(2)Ag(μ-mes)(2)(μ-dppy)]ClO(4) (2b) resulted luminescent in diluted DMSO solution at room temperature. Previously reported compound [Au(2)Cl(2)(μ-LL)] (LL dppy 5b) was also studied for comparative purposes. The antimicrobial activity of 1-5 and Ag[A] (A=ClO(4)(-), SO(3)CF(3)(-)) against gram-positive and gram-negative bacteria and yeast was evaluated. Most tested compounds displayed moderate to high antibacterial activity while heteronuclear Au(2)M derivatives with dppe (2a-4a) were the more active (minimum inhibitory concentration 10 to 1 μg mL(-1)). Compounds containing silver were ten times more active to gram-negative bacteria than the parent dinuclear compound 1a or silver salts. Au(2)Ag compounds with dppy (2b, 3b) were also potent against fungi.

Incidence and Risk Factors for Pulmonary Exacerbation Treatment Failures in Patients With Cystic Fibrosis Chronically Infected With Pseudomonas aeruginosa

Pulmonary exacerbations (PEx) are responsible for much of the morbidity and mortality associated with cystic fibrosis (CF). However, there is a paucity of data on outcomes in CF PEx and factors influencing outcomes. We reviewed all PEx in patients infected with Pseudomonas aeruginosa treated with parenteral antibiotics over 4 years at our center. Treatment failures were categorized a priori as those PEx requiring antibiotic regimen change, prolongation of therapy > 20 days because of failure to respond, an early recurrent event within < 45 days, or failure to recover lung function to > 90% of baseline FEV(1). A total of 101 patients were followed for 452 PEx. Treatment failures were observed in 125 (28%) of PEx; antibiotic regimen change was observed in 27 (6%), prolongation of therapy in 29 (6%), early recurrent events in 63 (14%), and failure to recover lung function to > 90% of baseline FEV(1) in 66 (15%). Demographic factors associated with one or more treatment failures per year included advanced airways disease, use of enteric feeds, CF-related diabetes, and CF liver disease but did not include female sex or F508del homozygosity. Increased treatment failure risk was associated with lower admission FEV(1) and increased markers of inflammation. At therapeutic completion, increased inflammatory markers correlated with treatment failure. Failure rates decreased with increasing number of active antimicrobial agents used based on in vitro susceptibility (zero, 28/65 [43%]; one, 38/140 [27%]; two, 59/245 [24%]; three, 0/2 [0%]; P = .02). One-fourth of PEx fail to respond adequately to initial management. Patient demographic and episode-specific clinical information can be used to identify individuals at increased risk of initial management failure.

Novel biologically active glycosides from the aerial parts of Cephalaria gazipashensis

Two new triterpene glycosides, 3-O-\beta-D-glucopyranosyl- (1\to 4)-\beta-D-xylopyranosyl-(1\to 3)-\alpha-L-rhamnopyranosyl- (1\to 3)-\alpha-L-arabinopyranosyl olean-12-ene 28-O-\beta -D- glucopyranosyl-(1\to 6)-\beta-D-glucopyranosyl ester (gazipashoside A, 1) and 3-O-\beta-D-glucopyranosyl-(1\to 4)-\beta-D-xylopyranosyl- (1\to 3)-\alpha-L-rhamnopyranosyl-(1\to 2)-\alpha-L-arabinopyranosyl hederagenin 28-O-\beta -D-glucopyranosyl-(1\to 6)-\beta-D-glucopyranosyl ester (gazipashoside B, 2), were isolated together with 11 known compounds including 7 triterpene, 1 flavonoid, and 3 iridoidal metabolites from the aerial parts of Cephalaria gazipashensis (Dipsacaceae). The structures of all compounds were elucidated by extensive evaluation of the spectroscopic data (1D- and 2D- NMR, HRESI-MS), as well as chemical examinations. The antimicrobial effects of all pure compounds were tested against gram-positive and gram-negative bacteria. The results indicated that the 2 new triterpene glycosides might be acceptable as antimicrobial active agents.

Nosocomial bloodstream infections due to Acinetobacter baumannii, Acinetobacter pittii and Acinetobacter nosocomialis in the United States

To compare the clinical features and antimicrobial susceptibilities of the clinically most important Acinetobacter species Acinetobacter baumannii, Acinetobacter pittii (formerly Acinetobacter genomic species 3) and Acinetobacter nosocomialis (formerly Acinetobacter genomic species 13TU). 295 Acinetobacter isolates collected prospectively from patients with bloodstream infections (BSI) in 52 US hospitals were identified to species level. Clinical and microbiological features were compared between species. A. baumannii (63%) was the most prevalent species, followed by A. nosocomialis (21%), and A. pittii (8%). Intravascular catheters (15.3%) and the respiratory tract (12.9%) were the most frequent sources of BSI. A higher overall mortality was observed in patients with A. baumannii BSI than in patients with BSI caused by A. nosocomialis and A. pittii (36.9% vs. 16.4% and 13.0%, resp., p<0.001). The most active antimicrobial agents as determined by broth microdilution were tigecycline (99.6% of isolates susceptible), colistin (99.3%), amikacin (98.5%), and imipenem (95.2%). 27 isolates (10.0%) were multi-drug resistant, all but one of these were A. baumannii. About one third of Acinetobacter BSI in our study were caused by A. nosocomialis or A. pittii. Patients with A. baumannii BSI had a less favorable outcome.

In vitro activity of tigecycline and comparators against Gram-negative pathogens isolated from blood in Europe (2004–2009)

Here we report on the antimicrobial resistance amongst Gram-negative isolates (excluding Acinetobacter spp.) collected from blood culture sources at European study sites as part of the global Tigecycline Evaluation and Surveillance Trial (T.E.S.T.) from the study start in 2004 until August 2009. All isolates were collected and tested for minimum inhibitory concentrations using Clinical and Laboratory Standards Institute methodology. Over the collection period, extended-spectrum β-lactamase (ESBL) production was recorded in 21.1% of Klebsiella pneumoniae, 2.6% of Klebsiella oxytoca and 11.3% of Escherichia coli, primarily in Croatia, Greece, Hungary, Italy, Poland, Romania and the Slovak Republic. ESBL rates stabilised amongst K. pneumoniae over 2006–2009, but doubled amongst E. coli in 2008–2009. The patterns of antimicrobial resistance changed accordingly for both organisms. Generally, Greece had the highest antimicrobial resistance for K. pneumoniae, Italy for E. coli, Serratia marcescens and Enterobacter spp., and Croatia for Pseudomonas aeruginosa. High resistance rates amongst K. pneumoniae were also seen in Croatia and Italy. Imipenem resistance amongst K. pneumoniae was reported exclusively in Greece (13.8%); amongst other Enterobacteriaceae, imipenem resistance was absent or low. Similarly, meropenem resistance was low amongst the Enterobacteriaceae except K. pneumoniae from Greece (42.6%). Across Europe, the most active antimicrobial agents against the Enterobacteriaceae were tigecycline, amikacin and the carbapenems, each with <10% resistance each year. Against the other antimicrobials, significant increases in non-susceptibility were reported for K. pneumoniae and E. coli, both important causative pathogens of bacteraemia.

Characterization of Fluoroquinolone-ResistantEscherichia coliCausing Septicemic Colibacillosis in Calves in Italy: Emergence of a Multiresistant O78 Clonal Group

An increased incidence of enrofloxacin-resistant Escherichia coli associated with septicemic colibacillosis in calves was observed recently in northern Italy. The aim of this study was to investigate this phenomenon. A total of 47 consecutive E. coli isolates exhibiting reduced susceptibility to enrofloxacin (intermediately resistant or resistant) causing septicemic colibacillosis in calves from 45 large-scale farms during 2006-2008, were studied. Phylogenetic group, antimicrobial agents susceptibility, and O serogroup were determined with randomly amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) typing, providing additional discrimination. All of the microorganisms carried resistance to two or more additional drugs, with the pattern fluoroquinolone-ampicillin-co-trimoxazole-tetracycline-gentamicin-thiamphenicol being the most represented (18/47; 38.3%). Plasmid-mediated extended-spectrum and AmpC β-lactamases and plasmid-mediated fluoroquinolone resistance genetic determinants were not detected. Third-generation cephalosporins emerged as the most active antimicrobial agents tested (97.9% of susceptible strains). Overall, 37 different RAPD profiles and 18 different O serogroups could be distinguished among the typeable strains, indicating a substantial heterogeneity and suggesting the occurrence of several independent selection events. However, approximately one-fourth (11/47) of the strains belonged to serogroup O78, and PFGE revealed that the great majority (7/11) of these were clonally related, indicating the selection of a O78 clonal group. This is the first report investigating the molecular epidemiology of fluoroquinolone-resistant E. coli in calves and describing the emergence of a fluoroquinolone-resistant E. coli clonal group in these animals.

Antioxidant, antimicrobial and anticancer activity of the lichens Cladonia furcata, Lecanora atra and Lecanora muralis

BackgroundThe aim of this study is to investigate in vitro antioxidant, antimicrobial and anticancer activity of the acetone extracts of the lichens Cladonia furcata, Lecanora atra and Lecanora muralis.MethodsAntioxidant activity was evaluated by five separate methods: free radical scavenging, superoxide anion radical scavenging, reducing power, determination of total phenolic compounds and determination of total flavonoid content. The antimicrobial activity was estimated by determination of the minimal inhibitory concentration by the broth microdilution method against six species of bacteria and ten species of fungi. Anticancer activity was tested against FemX (human melanoma) and LS174 (human colon carcinoma) cell lines using MTT method.ResultsOf the lichens tested, Lecanora atra had largest free radical scavenging activity (94.7% inhibition), which was greater than the standard antioxidants. Moreover, the tested extracts had effective reducing power and superoxide anion radical scavenging. The strong relationships between total phenolic and flavonoid contents and the antioxidant effect of tested extracts were observed. Extract of Cladonia furcata was the most active antimicrobial agent with minimum inhibitory concentration values ranging from 0.78 to 25 mg/mL. All extracts were found to be strong anticancer activity toward both cell lines with IC50 values ranging from 8.51 to 40.22 μg/mL.ConclusionsThe present study shows that tested lichen extracts demonstrated a strong antioxidant, antimicrobial and anticancer effects. That suggest that lichens may be used as as possible natural antioxidant, antimicrobial and anticancer agents to control various human, animal and plant diseases.

Cumulative clinical experience from over a decade of use of levofloxacin in urinary tract infections: critical appraisal and role in therapy

The treatment of urinary tract infections (UTIs) continues to evolve as common uropathogens increasingly become resistant to previously active antimicrobial agents. In addition, bacterial isolates, which were once considered to be either colonizers or contaminants, have emerged as true pathogens, likely related to the more complex array of settings where health care is now delivered. Even though the reliability of many antimicrobial agents has become less predictable, the fluoroquinolone group of agents has remained a frequent, if not the most often prescribed, antimicrobial therapy for almost all types of UTIs. Levofloxacin has taken its position at the top of the list as one of the most regularly administered fluoroquinolone agents given to patients with a suspected or proven UTI. The authors review the clinical experience of the use of levofloxacin over the past decade and suggest that the use of levofloxacin for the treatment of UTIs, although still fairly dependable, is perhaps not the best use of this important antimicrobial agent.

Risk factors and clinical characteristics of Stenotrophomonas maltophilia infections in neonates

The aim of this study was to review the risk factors and clinical, bacteriological, and epidemiological characteristics of Stenotrophomonas maltophilia infections in our neonatal intensive care unit. A retrospective matched case-control study was performed by comparing 23 cases of S maltophilia with 45 controls to identify the potential risk factors. To identify the case patients, the admission and medical records of patients in the neonatal intensive care unit and records from the Microbiology Department were reviewed between 2003 and 2008. Sepsis in two neonates (9%), conjunctivitis in two neonates (9%), and ventilator-associated pneumonia in 19 (82%) neonates were determined. Invasive-procedures, exposure to aminoglycoside and carbapenem, total parenteral nutrition, histamine 2 blockers, exposure to steroids, cholestasis, and duration of hospitalization were significantly associated with S maltophilia infections (p<0.05). On multivariate analysis, invasive procedures (odds ratio, 18.81) and duration of hospitalization (odds ratio, 1.06) were determined to be the risk factors for S maltophilia infection. The most active antimicrobial agent was trimethoprim/sulfamethoxazole (87%) for S maltophilia infection, and the mortality rate was 17%. Neonatologists should avoid from unnecessary invasive procedures and broad-spectrum antibiotics to reduce S maltophilia infections. Invasive procedures should be finished in the shortest time possible. Agent/factor-specific antibacterial treatment should be administered. Patients being discharged as early as possible will also reduce infection frequency. Stenotrophomonas maltophilia should be considered in patients with high Stenotrophomonas infection risk factors.

Supramolecular assemblies of rifampicin and cationic bilayers: preparation, characterization and micobactericidal activity

BackgroundCationic bilayers based on the inexpensive synthetic lipid dioctadecyldimethylammonium bromide (DODAB) have been useful as carriers for drug delivery, immunoadjuvants for vaccines and active antimicrobial agents.MethodsRifampicin (RIF) or isoniazid (ISO) interacted with DODAB bilayer fragments (BF) or large vesicles (LV). Dispersions were evaluated by dynamic light-scattering for zeta-average diameter (Dz) and zeta-potential (ζ) analysis; dialysis for determination of drug entrapment efficiency; plating and CFU counting for determination of cell viability of Mycobacterium smegmatis or tuberculosis, minimal bactericidal concentration (MBC) and synergism index for DODAB/drug combinations.ResultsDODAB alone killed micobacteria over a range of micromolar concentrations. RIF aggregates in water solution were solubilised by DODAB BF. RIF was incorporated in DODAB bilayers at high percentiles in contrast to the leaky behavior of ISO. Combination DODAB/RIF yielded MBCs of 2/2 and 4/0.007 μg/mL against Mycobacterium smegmatis or Mycobacterium tuberculosis, respectively. Synergism indexes equal to 0.5 or 1.0, indicated synergism against the former and independent action, against the latter species.ConclusionsIn vitro, DODAB acted effectively both as micobactericidal agent and carrier for rifampicin. The novel assemblies at reduced doses may become valuable against tuberculosis.

Detection of OXA‐type carbapenemases and integrons among carbapenem‐resistant Acinetobactor baumannii in a Teaching Hospital in China

The increasing trend of carbapenem-resistance (CR) and multi-drug resistance (MDR) in A. baumannii worldwide has limited the therapeutic effectiveness of antibiotic therapy. The study was conducted to determine the prevalence of carbapenemases and integrons among the isolates of imipenem-resistant A. baumannii (IRAB). A total of 71 non-repetitive imipenem- resistant A. baumannii isolates were collected and tested for susceptibility to 17 antimicrobials. The modified Hodge test and EDTA-disc synergy test were performed for the screening of carbapenemases and metallo-β -lactamases (MBLs) production, respectively. Isolates were then subjected to multiplex-PCR targeting genes encoding for OXA-type carbapenemase, MBLs and integrases. Random amplified polymorphic DNA (RAPD) genotyping was performed to assess genetic relatedness. All isolates exhibited multi-drug resistant phenotype. Colistin was the most active antimicrobial agent tested. Seventy-one isolates (100%) demonstrated positive in the modified Hodge test. Thirty-nine isolates showed positive in the EDTA-disc synergy test, however, no MBL genes were detected. All strains possessed a bla(OXA-51) -like gene. The co-exis-tence of bla(OXA-51) -like/bla(OXA-23) -like/intI1, bla(OXA-51) -like/bla(OXA-23) -like, bla(OXA-51) -like/bla(OXA-24) -like was detected in 91.6% (n = 65), 5.6% (n = 4), 2.8% (n = 2), respectively. Analysis of the genetic con-text of bla(OXA-23) showed the presence of ISAba1 upstream of bla(OXA-23) . No ISAba1 was detected upstream of bla(OXA-51) . Two different gene cassettes were found in these strains, and a high prevalence of aacA4, aadA1 and catB8 genes was observed. RAPD of 71 isolates showed 7 genotypes. The strains were mainly recovered from patients in intensive care unit, neurosurgery and department of respiratory disease. These findings show that multi-drug resistance in A. baumannii is a common problem. This study also shows a high distribution of bla(OXA-23) -like and intI1 genes in imipenem-resistant A. baumannii isolates. The clonal spread played an important role in the increase of OXA-23 producing IRABs in the hospital environment.

Identification and distribution of antimicrobial- active marine bacteria along the heavily polluted coastal area of Alexandria, Egypt

The main objective of the present study was to screen, identify and to study distribution of marine bacteria with antimicrobial activity in the heavily polluted coast of Alexandria, Egypt. The highest bacterial density positively correlated with pollution strength and localized in the central part of the coast in both water and sediments. Thirty four active bacterial isolates dominating the water and sediments of six highly polluted sites during winter and spring were identified. The most promising active bacterial strain was molecularly characterized as Enterococcus S29. Results confirmed the superiority and richness of the local coastal environment of Alexandria with active bacteria consisting up to 30.43% of sea water and 36.36% of sediments populations respectively at specific sites. These results provide an excellent potential for manipulating such natural and renewable source in the production of new and active antimicrobial agents for pharmaceutical applications.

Phytochemical properties and in-vitro antifungal activity of Senna alata Linn. crude stem bark extract

This study reports on the phytochemical properties and in-vitro antifungal activity of Senna alata Linn. crude stem bark extract on clinical test dermatophytes. The studies on the antifungal activities of crude extracts of Senna alata L. stem bark were carried out. The test was conducted on dermatophytes, which included clinical dermatophytes of the genera Trichophyton, Microsporum and Epidemophyton. These fungi are the causative agents of various types of dermotophytosis that attack various parts of the body and tend to the following conditions, Tinea capilis, Tinea cruris, Tinea corporis and Tinea pedis. The results obtained showed that the crude extract of the stem bark of Senna alata L. exhibited marked antifungal effects on Microsporum canslaslomyces, Trichophyton verrucosum,Trichophyton mentagrophytes, and Epidermophyton floccosum. The crude extract showed the highest inhibition on T. verrucosum and E. floccosum with 21.00 and 20.05 mm zones of inhibition respectively. The Minimum Inhibitory Concentration (MIC) was also performed and the result showed that the MIC of Senna alata L. crude stem bark extract on all the tested dermatophytes was 5.0 mg/mL. The result showed that the crude extract ofS. alata L stem bark was fungicidal for all tested dermatophytes at concentration of 10.00 and 5.00 mg/mL except E. floccosum which was only fungicidal at concentration of 10.00 mg/mL. The crude extract of S. alata L. stem bark screening showed vary degree of activities against all the tested dermatophytes. Phytochemical analysis revealed the presence of important secondary metabolites (tannins, steroids, alkaloids, anthraquinones, terpenes, carbohydrates and saponins) in the plant thus indicating the therapeutic potentials of the plant. It is concluded that S. alata L. stem bark could be a potential source of active antimicrobial agents, and a detailed assessment of its in vivo potencies and toxicological profile is therefore advocated. Key words: Antifungal activity, crude extract, dermatophytes, phytochemicals, Senna alataL., stem bark, MIC.

Purification and Characterization of an Antimicrobial Peptide, Insect Defensin, From Immunized House Fly (Diptera: Muscidae)

The house fly, Musca domestica L. (Diptera: Muscidae), is involved in phoretic movement of pathogenic agents, but it has a very efficient defense mechanism against infection. It is believed that antimicrobial peptides play a significant role in the defense system of the house fly. Here, we isolated a peptide from the immunized house fly pupae, measured its molecular mass (3987.6 Da) by matrix-assisted laser desorption ionization/time of flight-mass spectrometry, and determined its amino acid sequence by using the Procise Protein Sequencing System (Applied Biosystems, Foster City, CA). The peptide was confirmed as a member of the insect defensin family. It displayed high activity against gram-positive bacteria but lower activity against gram-negative bacteria and fungi. Reverse transcription-polymerase chain reaction showed that the house fly defensin gene was constitutively expressed in naive pupae and strongly up-regulated after immunization. House fly defensin is an amphiphilic peptide with a structure similar to that of the CSalphabeta scaffold of insect defensin A from the flesh fly, Phormia terraenovae Robineau-Desvoidy. To our knowledge, this is the first isolated and characterized house fly antimicrobial peptide, and our work may provide useful information for developing pharmacologically active antimicrobial agents.

Clinical Outcome of Mycobacterium abscessus Infection and Antimicrobial Susceptibility Testing

Mycobacterium abscessus is the most resistant and rapidly growing mycobacterium and causes a wide range of clinical infectious diseases. The relationship between antimicrobial susceptibility and clinical outcome needs to be further evaluated. Forty M. abscessus isolates were obtained from clinical specimens of 40 patients at the Taichung Veterans General Hospital from January 2006 to December 2008. Antimicrobial susceptibility testing was performed using the broth microdilution method according to the recommendations of the National Committee for Clinical Laboratory Standards. The clinical manifestations and outcomes were reviewed from medical records. Twenty-two patients were diagnosed with M. abscessus infection. Cough (86.3%), hemoptysis (31.8%) and fever (18.1%) were the most common symptoms. The radiographic findings included reticulonodular opacities (50.0%), consolidation (31.8%) and cavitary lesions (18.1%). The 40 isolates were susceptible to amikacin (95.0%), cefoxitin (32.5%), ciprofloxacin (10.0%), clarithromycin (92.5%), doxycycline (7.5%), imipenem (12.5%), moxifloxacin (22.5%), sulfamethoxazole (7.5%) and tigecycline (100%). The rate of treatment failure was 27.3% at the end of the 12(th) month after the start of treatment, although these patients were treated with a combination of clarithromycin and other antimicrobial agents. M. abscessus is naturally susceptible to clarithromycin and amikacin, variably susceptible to cefoxitin and imipenem, and resistant to most other antimicrobial drugs. Combination therapy with clarithromycin, amikacin and other active antimicrobial agents may lead to clinical improvement; however, the rate of treatment failure is still high.

Unraveling the Hydroxypropionaldehyde (HPA) System: An Active Antimicrobial Agent against Human Pathogens

The hydroxypropionaldehyde (HPA) system is a natural defense system synthesized by the probiotic bacterium Lactobacillus reuteri. To elucidate which of the molecules composing the HPA system (3-hydroxypropionaldehyde (3-HPA), reuterin (HPA dimer), and HPA hydrate) is responsible for the potent antimicrobial activity in biological systems, a combination of biochemical, genetic, and proteomic assays was used. The HPA system reacts with sulfhydryl-containing compounds such as cysteine and reduced glutathione (GSH) in solution. In situ, GSH knock-out Escherichia coli is significantly more susceptible to HPA-mediated cell death than E. coli wild type; GSH supplementation protects either bacteria from HPA attack. Proteomic analysis of HPA-treated bacteria ( Haemophilus influenzae ) revealed induction of redox- and heat shock-related proteins. A new antimicrobial mechanism of HPA is proposed, whereby the activity of HPA leads to depletion of free SH- groups in GSH and proteins through the action of 3-hydroxypropionaldehyde, causing an imbalance of the cellular redox status, ultimately resulting in cell death.

Active antimicrobial agents as components for naturally occurring biodegradable polymers

Schiff base polymer [P-4-HMPIMP] of 4-[(4-hydroxy-2-methylphenyl)imino-methyl]-2,6-dimethylphenol (4-HMPIMP) was synthesized via oxidative poly condensation reaction in an alkaline medium with using NaOCl, H2O2 or air as oxidants. The structures of the synthesized monomer and polymer were confirmed by FT-IR, UV-Vis, NMR and elemental analysis, while their properties were characterized by TG-DTA, size exclusion chromatography (SEC) and solubility tests. The influence of the type of oxidizer on Mn, Mw and PDI values was also studied. Conductivity measurements of the polymer was done by electrometer using four point probe technique and the electrochemical and optical band gap values of both 4-HMPIMP and P-4-HMPIMP were determined by using cyclic voltammetry and UV-Vis measurements. Conductivity and band gap values indicate that this polymer is a typical semiconductor.

Screening of herbal extracts against multi‐drug resistant Acinetobacter baumannii

Antibiotic resistance is increasing resulting in a decreasing number of fully active antimicrobial agents available to treat infections with multi-drug resistant (MDR) bacteria. Herbal medicines may offer alternative treatment options. A direct inoculation method simulating the standard disc diffusion assay was developed to determine in vitro antimicrobial activity of sixty herbal extracts against MDR-Acinetobacter baumannii (A. baumannii). Eighteen herbal extracts inhibited MDR-A. baumannii on agar plates, although the magnitude and quality of bacterial inhibition differed considerably among the antibacterial herbal extracts. Next, minimal inhibitory concentration (MIC) of these antibacterial herbal extracts was calculated using a broth microdilution assay. For most herbal extracts, the larger the zone of inhibition on agar plates, the lower the MIC. In general, hetero-resistance on agar plates correlated with higher MIC. The skip well phenomenon was seen with two herbal extracts. In conclusion, 30% of the screened herbal extracts demonstrated in vitro antibacterial activity against MDR-A. baumannii using similar rigorous testing methods as those commonly employed for assessing antimicrobial activity of synthetic antibacterial agents. Characterization of a specific compound conferring this antibacterial activity of the herbal extracts may help to identify novel antimicrobial agents active against highly resistant bacteria.

The Viriato Study

The Viriato study is a prospective, multicentre laboratory-based surveillance study of antimicrobial susceptibility in which 30 microbiology laboratories throughout Portugal are asked to isolate, identify and submit to a central laboratory for testing Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis responsible for community-acquired lower respiratory tract infections and Streptococcus pyogenes from tonsillitis. To monitor changes in antimicrobial resistance patterns of these frequent respiratory pathogens. Susceptibility was determined by disk diffusion (Kirby-Bauer) or using Etest strips following the Clinical and Laboratory Standards Institute guidelines. From 1999 to 2007 over 13 900 isolates were analysed. Among S. pneumoniae penicillin non-susceptibility decreased from 25% in 1999 to 18% in 2007 (p = 0.002) but resistance to macrolides showed a steady increase, reaching 20% in the last 6 years. Resistance to amoxicillin and the quinolones remained stable and very low (1-2%) throughout the study period. Antimicrobial resistance among H. influenzae and M. catarrhalis remained stable. The most significant resistance was to ampicillin, of 10-12% and greater than 80%, respectively, as a result of the production of beta-lactamases. Macrolide resistance among S. pyogenes was stable during 1999-2003 (20-23%) but after 2003 there was a steady decline in resistance, which in 2007 reached 10%. The Viriato surveillance study showed that penicillin remains the most active antimicrobial agent against S. pyogenes causing tonsillitis, and amoxicillin-clavulanate and the quinolones are the most active in vitro simultaneously against S. pneumoniae, H. influenzae and M. catarrhalis responsible for community-acquired lower respiratory tract infections in Portugal.

Nosocomial imipenem-resistant Acinetobacter baumannii infections: Epidemiology and risk factors

The incidence, clinical characteristics, risk factors, antimicrobial susceptibility, and outcomes of nosocomial imipenem-resistant A. baumannii (IRAB) infections during a 5-y period (2003-2007) were retrospectively analyzed. A total of 720 patients with 925 episodes of A. baumannii infection were included in the study. A. baumannii infections were seen mostly in intensive care units. The incidence was 6.2 per 1000 admissions. The most common infections were pneumonias and bloodstream infections. Imipenem resistance among Acinetobacter strains increased significantly each y of the study (from 43.3% to 72.9%). Mortality was related to the presence of imipenem resistance, stay in intensive care unit, female gender, old age, and pneumonia. Haemodialysis, malignancy, and mechanical ventilation were significant risk factors for IRAB infections. Imipenem resistance was higher in strains isolated from patients with pneumonia. IRAB strains showed higher resistance rates to other antibiotics than imipenem-susceptible strains. The most active antimicrobial agents against A. baumannii were cefoperazone-sulbactam and netilmicin. The incidence of A. baumannii infections and imipenem resistance increased during the study period. IRAB infections should be considered in patients on mechanical ventilation and haemodialysis and in patients with malignancies.