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Introduction: Due to an upsurge in antibiotic-resistant infections and lack of therapeutic options, new approaches are needed for treatment. Honey may be one such potential therapeutic option. We investigated the susceptibility of hospital acquired pathogens to four honeys from Wisconsin, United States, and then determined if the antibacterial effect of each honey against these pathogens is primarily due to the high sugar content. Methods: Thirteen pathogens including: four Clostridium difficile, two Methicillin-resistant Staphylococcus aureus, two Pseudomonas aeruginosa, one Methicillin-Susceptible Staphylococcus aureus, two Vancomycin-resistance Enterococcus, one Enterococcus faecalis and one Klebsiella pneumoniae were exposed to 1-50% (w/v) four Wisconsin honeys and Artificial honey to determine their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using the broth dilution method. Results: Buckwheat honey predominantly exhibited a bactericidal mode of action against the tested pathogens, and this varied with each pathogen. C. difficile isolates were more sensitive to the Wisconsin buckwheat honey as compared to the other pathogens. Artificial honey at 50% (w/v) failed to kill any of the pathogens. The high sugar content of Wisconsin buckwheat honey is not the only factor responsible for its bactericidal activity. Conclusion: Wisconsin buckwheat honey has the potential to be an important addition to therapeutic armamentarium against resistant pathogens and should be investigated further. [ABSTRACT FROM AUTHOR]

Elizabethkingia Meningoseptica is a gram-negative bacillus that is known to cause meningitis in premature neonates, and a variety of infections in immunocompromised adults. We are presenting the case of a 37 year old patient with an ileal perforation, in whom an ileostomy was done. He was on a ventilator for eleven days in the ICU and he developed septicaemia due to this organism. It is important that clinicians consider Elizabethkingia Meningoseptica also as a rare cause of septicaemia. The patient was successfully treated with levofloxacin, with a good clinical outcome.

Methicillin resistant staphylococcus aureus (MRSA) is a predominantly nosocomial pathogen which its prevalence has increased worldwide over the past three decades."nMaterials and Methods: This is a descriptive cross-sectional study. The following study is designed for determination of prevalence of MRSA infection through measurement of MICs of S. aureus isolates in Imam Khomeini Hospital (a teaching hospital) from November 2001 to January 2003. A total number of 402 specimens were isolated and specified as S. aureus by Imam Khomeini microbiology lab. Demographic and clinical data and results of MIC were analysed by Epilnfo 6 software."nResults: During the study, staphylococcus aureus was isolated from 402 patients that 187 (46.5%) of isolates were MRSA and 215 (53.5%) were MSSA. Of 402 patients, 254 (63.2%) were male and 148 (36.8%) were female. The difference of the prevalence of MRSA between males and females was not statistically significant (p= 0.09). The difference in mean age in MRSA and MSSA groups was not statistically significant (p= 0.55). In the age group of < 1 month, the prevalence of MRSA infection was significantly higher than other groups (P= 0.01)."nConclusion: In this study, the prevalence of MRSA infection was increased, statistically significant in the presence of such factors as sepsis, longer duration of hospitalization, hospital- acquired infection, history of invasive procedure, history of antimicrobials used in the past 3 months and type of administered antimicrobial (s), history of hospitalization in the preceding year, certain underlying diseases, type of admission ward, type of infection, type of specimen and type of administered antimicrobials for treatment. Surprisingly, the prevalence of MRSA infection in IV drug user group was low that was statistically significant (p< 0.0001). In this study, there was no statistically significant difference in outcome between MRSA infected and MSSA infected patients. Based on results of this study, prevalence of infections due MRSA in this hospital are high, therefore addition to diagnosis of risk factors of MRSA infections, application of standard infection control practices should be done for limitation of these infections.

Klebsiella pneumoniae is a nosocomial pathogen which naturally secretes lipopolysaccharide (LPS) and cell envelope associated proteins into the environment through the production of outer membrane vesicles (OMVs). The loss of the LPS O antigen has been demonstrated in other bacterial species to significantly alter the composition of OMVs. Therefore, this study aimed to comprehensively analyze the impact of O antigen loss on the sub-proteomes of both the outer membrane and secreted OMVs from K. pneumoniae . As determined by LC–MS/MS, OMVs were highly enriched with outer membrane proteins involved in cell wall, membrane, and envelope biogenesis as compared to the source cellular outer membrane. Deletion of wbbO , the enzyme responsible for O antigen attachment to LPS, decreased but did not eliminate this enrichment effect. Additionally, loss of O antigen resulted in OMVs with increased numbers of proteins involved in post-translational modification, protein turnover, and chaperones as compared to secreted vesicles from the wild type. This alteration of OMV composition may be a compensatory mechanism to deal with envelope stress. This comprehensive analysis confirms the highly distinct protein composition of OMVs as compared to their source membrane, and provides evidence for a selective sorting mechanism that involves LPS polysaccharides. These data support the hypothesis that modifications to LPS alters both the mechanics of protein sorting and the contents of secreted OMVs and significantly impacts the protein composition of the outer membrane. [ABSTRACT FROM AUTHOR]

Acinetobacter nosocomialis is an important nosocomial pathogen that causes a variety of opportunistic infections; however, pathogenesis of this microorganism has not yet been characterized. The aim of this study was to investigate the secretion of outer membrane vesicles (OMVs) from A. nosocomialis and to determine their cytotoxic effects and their ability to induce inflammatory responses both in vitro and in vivo by using human epithelial HEp-2 cells and a mouse model, respectively. A. nosocomialis ATCC 17903 T secreted spherical OMVs when cultured in vitro . Proteomic analysis revealed that 147 different proteins were associated with A. nosocomialis OMVs and virulence-associated proteins, such as outer membrane protein A (OmpA), CsuA, CsuC, CsuD, PilW, hemolysin, and serine protease, were identified. A. nosocomialis OMVs were cytotoxic to HEp-2 cells. These vesicles also induced the expression of pro-inflammatory cytokine genes in the HEp-2 cells. Early inflammatory responses, such as congestion and focal neutrophilic infiltration, were observed in the lungs of mice injected with A. nosocomialis OMVs. In conclusion, A. nosocomialis OMVs are important secretory nanocomplexes that induce cytotoxicity of epithelial cells and host inflammatory responses, which may contribute to the pathogenesis of A. nosocomialis . [ABSTRACT FROM AUTHOR]

Many antagonistic interactions between hosts and their parasites result in coevolution. Although coevolution can drive diversity and specificity within species, it is not known whether coevolutionary dynamics differ among functionally similar species. We present evidence of coevolution within simple communities of Pseudomonas aeruginosa PAO1 and a panel of bacteriophages. Pathogen identity affected coevolutionary dynamics. For five of six phages tested, time-shift assays revealed temporal peaks in bacterial resistance and phage infectivity, consistent with frequency-dependent selection (Red Queen dynamics). Two of the six phages also imposed additional directional selection, resulting in strongly increased resistance ranges over the entire length of the experiment (ca. 60 generations). Cross-resistance to these two phages was very high, independent of the coevolutionary history of the bacteria. We suggest that coevolutionary dynamics are associated with the nature of the receptor used by the phage for infection. Our results shed light on the coevolutionary process in simple communities and have practical application in the control of bacterial pathogens through the evolutionary training of phages, increasing their virulence and efficacy as therapeutics or disinfectants. [ABSTRACT FROM AUTHOR]

Abstract: We describe three types of Pseudomonas aeruginosa pneumonia. Case 1. P. aeruginosa was isolated from the blood and sputum of a 29-year-old male non-smoker who developed severe community-acquired pneumonia (CAP). Piperacillin was initially effective, but fever and lobular pneumonia with cavities developed seven days after discharge. Intravenous piperacillin/tazobactam and tobramycin were administered for four weeks, followed by oral ciprofloxacin for two weeks. He finally recovered, but developed recurrent CAP due to P. aeruginosa despite appropriate antibiotic therapy and immunocompetent status. Case 2. P. aeruginosa was isolated from the blood and sputum of a 57-year-old woman with renal cancer who developed hospital-acquired pneumonia (HAP) after surgical treatment. She recovered after meropenem administration for four weeks. Case 3. A 67-year-old woman with systemic sclerosis and malignant lymphoma who was followed up on an outpatient basis underwent immunosuppressive therapy. Thereafter, she developed pneumonia and was admitted to our institution where P aeruginosa was isolated from blood and sputum samples. Healthcare-associated pneumonia (HCAP) was diagnosed and effectively treated with tobramycin and ciprofloxacin. P. aeruginosa is not only a causative pathogen of HAP and HCAP, but possibly also of CAP. [Copyright &y& Elsevier]

The article discusses the extracellular enzyme profiling of Stenotrophomonas maltophilia clinical isolates. Stenotrophomonas maltophilia is still defined as an organism of limited pathogenicity. It has risen in prominence as a nosocomial pathogen. Very little is known about the extracellular enzymes profile, pigment production and motility patterns of Stenotrophomonas maltophilia which may have potential roles in pathogenesis.

Methicillin resistantStaphylococcus aureus(MRSA) is a very common nosocomial pathogen and in this study the occurrence of MRSA infections and their susceptibility pattern were tested. A total of 50 MRSA strains were collected from pus samples. All isolates were found to be sensitive to vancomycin and imipenem. Further minimum inhibitory concentration (MIC) of vancomycin was performed by agar dilution method, 24 % of the strains were inhibited at a concentration of 4 µg/ml and other 10 % at 8 µg/ml. It was observed that none of the isolates could tolerate 16 µg/ml and E-test also showed that 64 % of them were sensitive and 36 % were intermediately resistant. All the strains were tested for slime production by Congo red agar method and biofilm formation by microtitre plate method. Out of the total strains tested 58 % exhibited positive result for slime production and 70 % were biofilm positive. All the MRSA strains were tested for MIC with two types of honey which gave MIC of 30 %. Sarvodaya honey and Tulsi honey were chosen for the study. Further silver nanoparticles were synthesized using honey and the antimicrobial activities against the pathogens were checked. The study showed that honey has antibacterial activity against test organisms and may form an alternative therapy against certain bacteria. [ABSTRACT FROM AUTHOR]

Reasons for the rising clinical impact of the bacterium Enterococcus faecium include the species’ rapid acquisition of adaptive genetic elements. Here, we focused on the impact of recombination on the evolution of E. faecium. We used the recently developed BratNextGen algorithm to detect recombinant regions in the core genome of 34 E. faecium strains, including three newly sequenced clinical strains. Recombination was found to have a significant impact on the E. faecium genome: of the original 1.2 million positions in the core genome, 0.5 million were predicted to have been affected by recombination in at least one strain. Importantly, strains in one of the two major E. faecium clades (clade B), which contains most of the E. faecium human gut commensals, formed the most important reservoir for donating foreign DNA to the second major E. faecium clade (clade A), which contains most of the clinical isolates. Also, several genomic regions were found to mainly recombine in specific hospital-associated E. faecium strains. One of these regions (the epa-like locus) likely encodes the biosynthesis of cell wall polysaccharides. These findings suggest a crucial role for recombination in the emergence of E. faecium as a successful hospital-associated pathogen. [ABSTRACT FROM PUBLISHER]

Enterococcus faecalis , a human gastrointestinal tract commensal, is known to cause nosocomial infections. Interestingly, the pathogen's host colonization and persistent infections are possibly linked to its lifestyle changes from planktonic to sessile state. Also, the multidrug resistance and survival fitness acquired in the sessile stage of E. faecalis has challenged treatment regimes. This situation exists because of the critical role played by several root genes and their molecular branches, which are part of quorum sensing, aggregation substance, surface adhesions, stress-related response and sex pheromones in the sessile state. It is therefore imperative to decode the hidden agenda of E. faecalis and understand the significant factors influencing biofilm formation. This would, in turn, augment the development of novel strategies to tackle E. faecalis infections.

Abstract: Klebsiella pneumoniae MGH78578 contains ∼500 uncharacterized signaling proteins and in this study, we characterized the biological functions of a novel eukaryotic-like serine/threonine kinase; ESTK (KpnK). Studies demonstrated that KpnK undergoes autophosphorylation within the pH range 7.0–7.5 at 37°C in a time- and concentration- dependent manner, with Mn2+ as its cofactor. The ΔkpnK mutant exhibited higher sensitivity to gastrointestinal and oxidative stresses. Deletion of kpnK resulted in a two to threefold increased susceptibility towards imipenem, cefepime, ceftriaxone and ceftazidime. Our study has provided overall evidence for the involvement of ESTK in regulating bacterial physiology, stress response and drug resistance. This report has unmasked the occurrence of Ser/Thr kinase mediated signaling for the first time in K. pneumoniae. Structured summary of protein interactions: KpnK phosphorylates KpnK by protein kinase assay (View interaction). [Copyright &y& Elsevier]

Acinetobacter baumannii is a nosocomial bacterial pathogen, and infections attributed to this species are further complicated by a remarkable ability to acquire antimicrobial resistance genes and to survive in a desiccated state. While the antibiotic resistance and biofilm formation of A. baumannii is well-documented, less is known about the virulence attributes of this organism. Recent studies reported A. baumannii strains display a motility phenotype, which appears to be partially dependent upon Type IV pili, autoinducer molecules, and the response to blue light. In this study, we wanted to determine the prevalence of this trait in genetically diverse clinical isolates, and any additional required factors, and environmental cues that regulate motility. When strains are subjected to a wide array of stress conditions, A. baumannii motility is significantly reduced. In contrast, when extracellular iron is provided or salinity is reduced, motility is significantly enhanced. We further investigated whether the genes required for the production of lipopolysaccharide ( lpsB) and K1 capsule ( epsA/ ptk) are required for motility as demonstrated in other Gram-negative bacteria. Transposon mutagenesis resulted in reduced motility by the insertion derivatives of each of these genes. The presence of the parental allele provided in trans, in the insertion mutant background, could only restore motility in the lpsB mutant. The production of core LPS directly contributes to the motility phenotype, while capsular polysaccharide may have an indirect effect. Further, the data suggest motility is regulated by extracellular conditions, indicating that A. baumannii is actively sensing the environment and responding accordingly. [ABSTRACT FROM AUTHOR]

The outer membrane protein A (OmpA) plays important roles in anchoring of the outer membrane to the bacterial cell wall. The C-terminal periplasmic domain of OmpA (OmpA-like domain) associates with the peptidoglycan (PGN) layer noncovalently. However, there is a paucity of information on the structural aspects of the mechanism of PGN recognition by OmpA-like domains. To elucidate this molecular recognition process, we solved the high-resolution crystal structure of an OmpA-like domain from Acinetobacter baumannii bound to diaminopimelate (DAP), a unique bacterial amino acid from the PGN. The structure clearly illustrates that two absolutely conserved Asp271 and Arg286 residues are the key to the binding to DAP of PGN. Identification of DAP as the central anchoring site of PGN to OmpA is further supported by isothermal titration calorimetry and a pulldown assay with PGN. An NMR-based computational model for complexation between the PGN and OmpA emerged, and this model is validated by determining the crystal structure in complex with a synthetic PGN fragment. These structural data provide a detailed glimpse of how the anchoring of OmpA to the cell wall of gram-negative bacteria takes place in a DAP-dependent manner. [ABSTRACT FROM AUTHOR]

Background: The aim of this study was to investigate the susceptibility profiles to disinfectants and antimicrobial agents of 283 non-repetitive Acinetobacter clinical isolates obtained in 97 Japanese hospitals in March 2002. Methods: Susceptibility profiles of the above isolates to four disinfectants, six antimicrobial agents and two dyes were investigated. MICs were measured by the agar dilution method recommended by the CLSI (formerly NCCLS). MBC measurements and time-kill assays were performed using a slightly modified quantitative suspension test based on the European Standard EN 1040. Results: No evident resistance to disinfectants was seen among the 283 strains of Acinetobacter spp. isolated in 2002, but the MIC90s of chlorhexidine gluconate, benzalkonium chloride and alkyldiaminoethylglycine hydrochloride were 50, 50 and 400 mg/L, respectively. Interestingly, the MICs of alkyldiaminoethylglycine hydrochloride and benzethonium chloride for four and three clinical isolates, respectively, reached 800 mg/L (approximately half the in-use concentration). The MBCs for the 28 disinfectant reduced susceptibility (DRS) isolates, for which the MICs of at least one of the four disinfectants tested were higher than the MIC90, were comparable to those for susceptible isolates, in general; however, significant differences (P<0.01) were observed between disinfectant-susceptible and DRS isolates in the time-kill assays of chlorhexidine gluconate, benzalkonium chloride and benzethonium chloride. Furthermore, DRS isolates tended to demonstrate multiresistance profiles to ceftazidime, ciprofloxacin and amikacin (P<0.05). Conclusions: Since several Acinetobacter clinical isolates have developed augmented resistance to multiple antimicrobials and disinfectants, it is worth checking the susceptibility to disinfectants if multidrug-resistant Acinetobacter spp. are recurrently isolated clinically. [ABSTRACT FROM AUTHOR]

Background & objectives: Cetrimide is a monoeationic surfactant, commonly used for disinfection of hospital floors, equipments, for cleansing of burns and wounds, hand wash, etc. We evaluated whether antibiotic resistant (AR) Escherichia coli isolates from hospital settings (nosocomial pathogens) show any evidence of significant reduction in their susceptibility to cetrimide. Also the response of AR E. coli (nosoeomial pathogens) to the action of cetrimide was assessed by studying the ultra structural changes induced using transmission electron microscopy (TEM). Methods: A total of 165 clinical samples were screened for isolation of E. coli. Eighty two (49.6%) samples were positive for E. coli Antibiotic susceptibility testing of E. coli isolates was carried out by Kirby Bauer method to isolate AR E. coli. The randomly selected AR E. coli isolate was treated with different concentrations of cetrimide and minimum inhibitory concentration (MIC) of cetrimide was determined by broth micro dilution method. This same isolate was used for performing time kill assay and TEM study. Results: The test E. coli isolate showed resistance to 12 different antibiotics. The MIC of cetrimide against ARE. coli was 312.5 µg/ml. The ultra cellular structural changes in cetrimide treated AR E. coli revealed vacuole formation, disaxilization of nuclear material, loss of cytoplasmic granularity, bleb formation and cell lysis. Conclusion: Ultra structural changes induced by the action of cetrimide revealed cell damaging changes in the AR E. coli to be dose and time dependent. The results showed that antibiotic resistance does not alter any change in susceptibility of E. coli to cetrimide, which was found to be still an effective disinfectant against a nosocomial pathogen E. coli. [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa is well-recognized as a nosocomial pathogen, which exhibits inherent drug resistance. In this study, the antibacterial activity of ethanol extracts of 58 Chinese herbal medicines used in Taiwan were tested against 89 nosocomial antibiotic resistant strains of Pseudomonas aeruginosa. The results gathered by the disc diffusion method showed that 26 out of the 58 herbal extracts exhibited antibacterial activity. Among the 26 herbal extracts, 10 extracts showed broad-spectrum antibacterial activities and were selected for further antibacterial property assay. The minimum inhibitory concentrations (MIC) of the active partition fractions ranged from 0.25 to 11.0 mg/L. The presence of flavonoid compounds in the active fractions of test herbal extracts was observed by the TLC-bioautography. The results from the time-kill assay revealed that most of the herbal extracts completely killed the test organisms within 4 hours. Exposure of the test strains to a sub-MIC level of the herbal extracts for 10 consecutive subcultures did not induce resistance to the active components. A combination of the active herbal fractions with antibiotics showed that one of the herbal medicines, the hexane fraction of Ramulus Cinnamomi, possessed a synergistic effect with tetracycline, gentamycin, and streptomycin. In conclusion, the tested Chinese medical herbs have the potential to be developed into natural antibiotics. This is the first evaluation for screening large amounts of medical plants against nosocomial antibiotic resistant bacteria in Taiwan. [ABSTRACT FROM AUTHOR]

Abstract: Infection by nosocomial pathogenic bacteria is increasingly becoming a major threat to the patients in the hospital. We have developed a diagnostic DNA microarray for the detection of two important nosocomial pathogens, Pseudomonas aeruginosa and Acinetobacter baumannii. The diagnostic DNA microarray contains the species-specific probes of 15mer oligonucleotides designed based on the sequences of 23S ribosomal DNA. The performance of DNA microarray in diagnosing P. aeruginosa and A. baumannii was evaluated using reference bacteria as well as clinical specimens such as blood, stool, pus, sputum, urine and cerebrospinal fluid. Using this DNA microarray, A. baumannii could be successfully detected in 11 out of 13 clinical specimens, thus giving the sensitivity of 84.6% with the specificity of 100% and the positive predictive value of 100%. P. aeruginosa could also be detected in 25 out of 26 clinical specimens, showing the sensitivity of 96.2%, the specificity of 100%, and the positive predictive value of 100%. These results suggest that two nosocomial pathogens, P. aeruginosa and A. baumannii, can be efficiently diagnosed by using the DNA microarray developed in this study. [Copyright &y& Elsevier]

Abstract: During a 53-month period (March 1994 to August 1998), 48 Corynebacterium striatum isolates recovered from clinical specimens were characterized. The organisms were identified by both phenotypic characteristics and 16S rRNA gene sequence analysis. Thirty-six (75%) were isolated from sputum/bronchial aspirates, 10 (21%) from wound exudates/pus, 1 (2%) from vaginal discharge, and 1 (2%) from an otorrheic specimen. All 48 patients had been hospitalized for treatment of an underlying disease and had received antibiotics previously. The C. striatum isolates were considered pathogenic based on their abundance within polymorphonuclear neutrophils and their dominant growth in culture. Sensitivities of isolates to 11 antibiotics were determined by broth microdilution. MIC90 values of the isolates were 1 μg/mL for vancomycin, 16 μg/mL for penicillin and ampicillin, 32 μg/mL for minocycline, and ≥32 μg/mL for cephalosporins, imipenem, ofloxacin, and macrolides. Restriction fragment-length polymorphism analysis with pulsed-field gel electrophoresis was used to determine the clonal identity. The pulse-field gel electrophoresis profiles revealed 14 distinct patterns with 20 subtypes. The isolates for the nosocomial outbreaks of C. striatum included 3 types (A, D, and E) with 4 subtypes (A1, A2, D2, and E). All 4 genotypes had broad-spectrum resistance to antimicrobial agents. Furthermore, type E strain isolated from 3 patients in the same ward was sensitive only to vancomycin. We conclude that C. striatum should be considered an emerging multidrug-resistant nosocomial pathogen in patients hospitalized for a prolonged period and/or in immunocompromised patients with such underlying conditions as cerebrovascular disease, pulmonary disease, diabetes, or malignancy. [Copyright &y& Elsevier]

Resistance of bacteria to antimicrobial agents is a worldwide concern. In Korea, resistant bacteria are more prevalent than in other industrialized countries. Methicillin-resistant Staphylococcus aureus, erythromycin-resistant Streptococcus pyogenes, penicillin non-susceptible pneumococci, β-lactamase-producing gonococci, extended-spectrum β-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae, class C β-lactamase-producing E. coli, fluoroquinolone-resistant E. coli, and aminoglycoside-resistant Acinetobacter baumannii and Pseudomonas aeruginosa are examples of resistant bacteria prevalent in Korea, and their presence suggests a high level of antimicrobial selective pressure and the nosocomial spread of resistant bacteria. Recently observed rapid increases in the incidence of vancomycin-resistant Enterococcus faecium and carbapenem-resistant P. aeruginosa present new threats in Korea. [ABSTRACT FROM AUTHOR]

Background: Recently, Acinetobacter has emerged as significant hospital pathogen, notoriously known to acquire antibiotic resistance to most of the commonly prescribed antimicrobials. Many risk factors are associated with Acinetobacter infections, especially in patients in intensive care unit (ICU). This study aims to isolate Acinetobacter from various clinical specimens and to determine its antimicrobial sensitivity pattern. Materials and Methods: Identification, speciation and antimicrobial sensitivity testing were performed using the standard microbiological techniques. Slime production was also tested by microtiter plate and tube method. Results: From the processed clinical specimens, 107 Acinetobacter strains (1.02%) were isolated of which 76 (0.74%) isolates were from general wards and 31 (11.96%) were from ICU. Significantly higher percentage of Acinetobacter strains was found in ICU compared with general wards (P < 0.05). Most common Acinetobacter infection was abscess. Infections were more common in males and were associated with major risk factors such as post-surgical, diabetes mellitus, catheterization, extended hospital stay and prolonged antibiotic usage. Acinetobacter baumanii was the most common species isolated to cause abscess, wound infection, etc. 62.61% and 28.97% isolates produced slime by microtiter plate and tube method. Imipenem was most sensitive drug followed by amikacin. Ceftazidime, cefotaxime, piperacillin were most resistant. 43.00% isolates were IPM resistant. A. baumanii was more resistant to commonly used antimicrobials. Conclusion: Acinetobacter nosocomial infections resistant to most antimicrobials have emerged, especially in ICU. Early identification and continued surveillance of prevalent organism will help prevent the spread of Acinetobacter in hospital environment.

Introduction: Ralstonia mannitolilytica is a rare opportunistic pathogen capable of causing a serious infection in immunocompromised patients. Our objective was to describe all cases of R. mannitolilytica bloodstream infection identified within 2 years at our tertiary care centre, focusing on clinical characteristics, risk factors, antibiotic sensitivity patterns, management and outcomes., Case Series: We compiled a descriptive case series including 14 non-duplicate R. mannitolilytica isolates obtained from bloodstream infection samples from the microbiology laboratory of a tertiary care centre from June 2019 to June 2021. All isolates were initially identified based on their morphological properties and biochemical reactions, and then underwent matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) examination for confirmation of identity. Antibiotic susceptibility testing was performed using the Kirby-Bauer disc diffusion method and Vitek 2. All 14 patients presented with symptoms of fever and/or chills, and a positive blood culture for R. mannitolilytica . After 48 h of incubation, no Ralstonia growth was reported from any of the current environmental or pharmaceutical water samples. Chemotherapy (9/14), mechanical ventilation (4/14), steroid use (2/14) and diabetes mellitus (1/14) were associated risk factors in our patients. The antibiotic sensitivity panel showed maximum resistance to aminoglycosides (64.3%) and no resistance to cefoperazone/sulbactum. Patients received treatment with cefoperazone/sulbactum and meropenem or ceftazidime. Thirteen patients recovered with antibiotic therapy and one patient succumbed to his illness., Conclusion: R. mannitolilytica can cause bloodstream infections in immunocompromised patients. It is likely to be missed or underreported due to lack of clinical awareness. MALDI-TOF MS is helpful in rapid identification. R. mannitolilytica is resistant to many routinely used antibiotics, including carbapenems., Competing Interests: The authors declare that there are no conflicts of interest., (© 2022 The Authors.)

Background: Acinetobacter baumannii is an important nosocomial pathogen causing high morbidity and mortality in immunocompromised patients with prolonged hospitalization. Multidrug-resistant A. baumannii infections are on the rise worldwide. Therefore, the discovery of an effective vaccine against this bacterium seems necessary as a cost-effective and preventive strategy., Methods: In this present study, 35 genomes of A. baumannii strains were considered, and the extracellular proteins were selected, maximally having one transmembrane helix with high adhesion probability and no similarity to host proteins, as immunogenic candidates using the web tool Vaxign. Subsequently, the role of these selected proteins in bacterial pathogenesis was investigated using VICMpred. Then, the major histocompatibility complex class II, linear B-cell epitopes, and conservation of epitopes were identified using the Immune Epitope Database, BepiPred, and Epitope Conservancy Analysis, respectively. Finally, the B-cell discontinuous epitopes of each protein were predicted using ElliPro and plotted on the three-dimensional structure (3D) of the proteins. The role of the unknown proteins was predicted using the STRING database., Results: In this study, eight acceptable immunogenic candidates, including FilF, FimA, putative acid phosphatase, putative exported protein, subtilisin-like serine protease, and three uncharacterized proteins, were identified in A. baumannii ., Conclusions: The results of the STRING database showed that these three uncharacterized proteins play a role in nutrition (heme utilization), peptide bond cleavage (serine peptidases), and cellular processes (MlaD protein). Extracellular proteins might play a catalyst role in the outer membrane protein-based vaccine of A. baumannii . Furthermore, this study proposed a list of potent immunogenic candidates of extracellular proteins., Competing Interests: Conflict of Interests: The authors have no conflict of interest to declare., (Copyright © 2022, Author(s).)

The prevalence of multidrug-resistant Gram-negative bacteria is a public health concern. Bacteriophages and bacteriophage-derived lytic enzymes have been studied in response to the emergence of multidrug-resistant bacteria. The availability of tRNAs and endolysin toxicity during recombinant protein expression is circumvented by codon optimization and lower expression levels using inducible pET-type plasmids and controlled cultivation conditions, respectively. The use of polyhistidine tags facilitates endolysin purification and alters antimicrobial activity. Outer membrane permeabilizers, such as organic acids, act synergistically with endolysins, but some endolysins permeate the outer membrane of Gram-negative bacteria per se. However, the outer membrane permeation mechanisms of endolysins remain unclear. Other strategies, such as the co-administration of endolysins with polymyxins, silver nanoparticles, and liposomes confer additional outer membrane permeation. Engineered endolysins comprising domains for outer membrane permeation is also a strategy used to overcome the current challenges on the control of multidrug-resistant Gram-negative bacteria. Metagenomics is a new strategy for screening endolysins with interesting antimicrobial properties from uncultured phage genomes. Here, we review the current state of the art on the heterologous expression of endolysin, showing the potential of bacteriophage endolysins in controlling bacterial infections. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii, a Gram-negative bacterium, is an important nosocomial pathogen. Colistin-resistant A. baumannii is becoming a new concern, since colistin is one of the last-line antibiotics for infections by carbapenem-resistant A. baumannii. From 452 carbapenem-resistant isolates collected in a teaching hospital in Taipei, Taiwan, we identified seven that were resistant to colistin. Carbapenem resistance in these isolates is attributed to the presence of carbapenemase gene blaOXA-23 in their genomes. Colistin resistance is presumably conferred by mutations in the sensor kinase domain of PmrB found in these isolates, which are known to result in modification of colistin target lipid A via the PmrB–PmrA–PmrC signal transduction pathway. Overexpression of pmrC, eptA, and naxD was observed in all seven isolates. Colistin resistance mediated by pmrB mutations has never been reported in Taiwan. One of the seven isolates contained three mutations in lpxD and exhibited an altered lipopolysaccharide profile, which may contribute to its colistin resistance. No significant difference in growth rates was observed between the isolates and the reference strain, suggesting no fitness cost of colistin resistance. Biofilm formation abilities of the isolates were lower than that of the reference. Interestingly, one of the isolates was heteroresistant to colistin. Four of the isolates were significantly more virulent to wax moth larvae than the reference. [ABSTRACT FROM AUTHOR]

Treatment of infections caused by multidrug-resistant (MDR) Gram-negative bacteria is challenging, a potential solution for which is the use of bacteriophage-derived lytic enzymes. However, the exogenous action of bacteriophage lysins against Gram-negative bacteria is hindered due to the presence of an impermeable outer membrane in these bacteria. Nevertheless, recent research has demonstrated that some lysins are capable of permeating the outer membrane of Gram-negative bacteria with the help of signal peptides. In the present study, we investigated the genomes of 309 bacteriophages that infect Gram-negative pathogens of clinical interest in order to determine the evolutionary markers of signal peptide-containing lysins. Complete genomes displayed 265 putative lysins, of which 17 (6.41%) contained signal-arrest-release motifs and 41 (15.47%) contained cleavable signal peptides. There was no apparent relationship between host specificity and lysin diversity. Nevertheless, the evolution of lysin genes might not be independent of the rest of the bacteriophage genome once pan-genome clustering and lysin diversity appear to be correlated. In addition, signal peptide- and signal-arrest-release-containing lysins were monophyletically distributed in the protein cladogram, suggesting that the natural selection of holin-independent lysins is divergent. Our study screened 58 (21.89%) out of 265 potential candidates for in vitro experimentation against MDR bacteria. [ABSTRACT FROM AUTHOR]

Background: Enterococci form a part of the normal flora of the intestinal tract, the oral cavity, and the vagina, but in recent times, they have become emerging nosocomial pathogens. Their increasing importance is largely due to their resistance to antimicrobials. The therapeutic failures in enterococcal infections are mainly due to the intrinsic as well as transferable drug resistance. The main aim of our study was to estimate the prevalence of the Enterococcus infection and to determine the antibiogram in a tertiary care hospital . Method : Enterococcus was isolated from a total of 5555 clinical samples like urine , pus, tissue, blood and body fluids during the period from January to December 2008.The isolates were speciated by using conventional biochemical tests (Facklam and Collins). The antibiotic susceptibility testing was done by the Kirby Bauer disc diffusion method. Confirmation of vancomycin susceptibility was done by the Epsilometer test (E test) to determine the Minimum Inhibitory Concentration (MIC). Result: From various clinical samples, 128 Enterococcus species were isolated in a period of one year and the rate of the infection was estimated to be 2.3%. Among the isolates, those of Enterococcus faecalis (E.faecalis) were 97(76%) and the remaining 31(24%) were of Enterococcus faecium (E.faecium). The maximum number of isolates were from pus 55(43%), followed by the isolates from urine 40(31%). The sensitivity pattern of these isolates showed an increased resistance to penicillin, ampicillin and ciprofloxacin. A High Level of Gentamicin Resistance (HLGR) was present in 60 (47% ) isolates of Enterococcus and 35(27%) isolates were intermediately sensitive to vancomycin by the Kirby Bauer disc diffusion method. All the intermediately sensitive isolates to vancomycin were further tested by the E test and they were found to be vancomycin sensitive. Conclusion: Various studies have shown an increase in the rate of infection and the antibiotic resistance in the Enterococcus species. There is also a change in the pattern of the Enterococcus infection, with an increase in the isolation rate of E. faecium and other non faecalis Enterococcus species. The Kirby Bauer disc diffusion method is not an accurate method for detecting the Vancomycin Resistant Enterococci (VRE). [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa high-risk clones are disseminated worldwide and they are common causative agents of hospital-acquired infections. In this review, we will summarize available data of high-risk P. aeruginosa clones from confirmed outbreaks and based on whole-genome sequence data. Common feature of high-risk clones is the production of beta-lactamases and among metallo-beta-lactamases NDM, VIM and IMP types are widely disseminated in different sequence types (STs), by contrast FIM type has been reported in ST235 in Italy, whereas GIM type in ST111 in Germany. In the case of ST277, it is most frequently detected in Brazil and it carries a resistome linked to blaSPM. Colistin resistance develops among P. aeruginosa clones in a lesser extent compared to other resistance mechanisms, as ST235 strains remain mainly susceptible to colistin however, some reports described mcr positive P. aeurigonsa ST235. Transferable quinolone resistance determinants are detected in P. aeruginosa high-risk clones and aac(6′)-Ib-cr variant is the most frequently reported as this determinant is incorporated in integrons. Additionally, qnrVC1 was recently detected in ST773 in Hungary and in ST175 in Spain. Continuous monitoring and surveillance programs are mandatory to track high-risk clones and to analyze emergence of novel clones as well as novel resistance determinants. [ABSTRACT FROM AUTHOR]

This study analyzed the genotype, antibiotic resistance, and biofilm formation of Acinetobacter baumannii strains and assessed the correlation between biofilm formation, antibiotic resistance, and biofilm-related risk factors. A total of 207 non-replicate multi-drug-resistant A. baumannii strains were prospectively isolated. Phenotypic identification and antimicrobial susceptibility testing were carried out. Isolate biofilm formation ability was evaluated using the tissue culture plate (TCP), Congo red agar, and tube methods. Clonal relatedness between the strains was assessed by enterobacterial repetitive intergenic consensus-PCR genotyping. Of the 207 isolates, 52.5% originated from an intensive care unit setting, and pan resistance was observed against ceftazidime and cefepime, with elevated resistance (99–94%) to piperacillin/tazobactam, imipenem, levofloxacin, and ciprofloxacin. alongside high susceptibility to tigecycline (97.8%). The Tissue culture plate, Tube method, and Congo red agar methods revealed that 53.6%, 20.8%, and 2.7% of the strains were strong biofilm producers, respectively, while a significant correlation was observed between biofilm formation and device-originating respiratory isolates (p = 0.0009) and between biofilm formation in colonized vs. true infection isolates (p = 0.0001). No correlation was detected between antibiotic resistance and biofilm formation capacity, and the majority of isolates were clonally unrelated. These findings highlight the urgent need for implementing strict infection control measures in clinical settings. [ABSTRACT FROM AUTHOR]

Prophages are often involved in host survival strategies and contribute toward increasing the genetic diversity of the host genome. Prophages also drive horizontal propagation of various genes as vehicles. However, there are few retrospective studies contributing to the propagation of antimicrobial resistance (AMR) and virulence factor (VF) genes by prophage. We extracted the complete genome sequences of seven pathogens, including ESKAPE bacteria and Escherichia coli from a public database, and examined the distribution of both the AMR and VF genes in prophage-like regions. We found that the ratios of AMR and VF genes greatly varied among the seven species. More than 70% of Enterobacter cloacae strains had VF genes, but only 1.2% of Klebsiella pneumoniae strains had VF genes from prophages. AMR and VF genes are unlikely to exist together in the same prophage region except in E. coli and Staphylococcus aureus , and the distribution patterns of prophage types containing AMR genes are distinct from those of VF gene-carrying prophage types. AMR genes in the prophage were located near transposase and/or integrase. The prophage containing class 1 integrase possessed a significantly greater number of AMR genes than did prophages with no class 1 integrase. The results of this study present a comprehensive picture of AMR and VF genes present within, or close to, prophage-like elements and different prophage patterns between AMR- or VF-encoding prophage-like elements. IMPORTANCE Although we believe phages play an important role in horizontal gene transfer in exchanging genetic material, we do not know the distribution of the antimicrobial resistance (AMR) and/or virulence factor (VF) genes in prophages. We collected different prophage elements from the complete genome sequences of seven species-Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter cloacae, and Escherichia coli-and characterized the distribution of antimicrobial resistance and virulence genes located in the prophage region. While virulence genes in prophage were species specific, antimicrobial resistance genes in prophages were highly conserved in various species. An integron structure was detected within specific prophage regions such as P1-like prophage element. Maximum of 10 antimicrobial resistance genes were found in a single prophage region, suggesting that prophages act as a reservoir for antimicrobial resistance genes. The results of this study show the different characteristic structures between AMR- or VF-encoding prophages.