Low Frequency Of Resistance Research Articles

Mode of Action and Heterologous Expression of the Natural Product Antibiotic Vancoresmycin

Antibiotics that interfere with the bacterial cytoplasmic membrane have long-term potential for the treatment of infectious diseases as this mode of action is anticipated to result in low resistance frequency. Vancoresmycin is an understudied natural product antibiotic consisting of a terminal tetramic acid moiety fused to a linear, highly oxygenated, stereochemically complex polyketide chain. Vancoresmycin shows minimum inhibitory concentrations (MICs) from 0.125 to 2 μg/mL against a range of clinically relevant, antibiotic-resistant Gram-positive bacteria. Through a comprehensive mode-of-action study, utilizing Bacillus subtilis reporter strains, DiSC3(5) depolarization assays, and fluorescence microscopy, we have shown that vancoresmycin selectively targets the cytoplasmic membrane of Gram-positive bacteria via a non-pore-forming, concentration-dependent depolarization mechanism. Whole genome sequencing of the producing strain allowed identification of the 141 kbp gene cluster encoding for vancoresmycin biosynthesis and a preliminary model for its biosynthesis. The size and complex structure of vancoresmycin could confound attempts to generate synthetic analogues. To overcome this problem and facilitate future studies, we identified, cloned, and expressed the 141 kbp biosynthetic gene cluster in Streptomyces coelicolor M1152. Elucidation of the mode-of-action of vancoresmycin, together with the heterologous expression system, will greatly facilitate further studies of this and related molecules.

EchinochloaResistance to Herbicides Continues to Increase in Arkansas Rice Fields

AbstractHerbicide-resistantEchinochloaspp. pose a significant threat to U.S. rice production. Two surveys were conducted to characterizeEchinochloaresistance to common rice herbicides and provide important demographic information on the populations in Arkansas: one was theEchinochloa Herbicide Resistance Confirmation Surveyconducted annually since 2006; the other was theEchinochloa Herbicide Resistance Demographics Surveyconducted since 2010. TheResistance Confirmation Surveyshowed that resistance to propanil (50%) was most prevalent, followed by quinclorac (23%), imazethapyr (13%), and cyhalofop (3%). Multiple resistance increased with time, with 27% of accessions being multiple-resistant, mostly to propanil+quinclorac (12%). The parallelResistance Demographics Surveytested resistance by species. Of the 264 accessions collected, 73% were junglerice, 14% were rough barnyardgrass, and 11% were barnyardgrass. Overall, this survey also showed resistance to propanil (53%) and quinclorac (28%) being most prevalent, with low frequencies of resistance to cyhalofop (12%) and imazethapyr (6%). Resistance to herbicides was less frequent with barnyardgrass (54%) and rough barnyardgrass (28%) than with junglerice (73%). Multiple resistance was most frequent with junglerice (33%) and least frequent with rough barnyardgrass (8%). Across both surveys, the resistance cases were clustered in the northeast and Grand Prairie regions of the state. Herbicide resistance amongEchinochloapopulations in rice fields is continuing to increase in frequency and complexity. This is a consequence of sequential selection with different major herbicide sites of action, starting with propanil followed by quinclorac and others.

A Hospital-based Case-control Study of Diarrhea in Children in Shanghai.

Prevention and control of childhood diarrhea is a global priority. We conducted a case-control study of childhood diarrhea in Shanghai. We prospectively recruited diarrheal children in an outpatient setting. Nondiarrheal controls were individually matched to cases. Rotavirus, norovirus and bacterial pathogens were examined. Clinical and epidemiologic data were obtained at enrollment and follow-up. Potential pathogens identified in 680 diarrheal cases and 680 controls were rotavirus (19.0% vs. 1.3%), norovirus (13.4% vs. 4.7%), nontyphoidal Salmonella (9.3% vs. 1.9%), enteropathogenic Escherichia coli (8.4% vs. 6.9%) and enteroaggregative Escherichia coli (7.2% vs. 6.2%) and Campylobacter (5.1% vs. 1.2%), enterotoxigenic Escherichia coli (1.2% vs. 0.6%), enterohemorrhagic Escherichia coli (0.3% vs. 0%) and Shigella (0.15% vs. 0%), respectively. The specificity and sensitivity of fecal leukocytes >5 per high-power field for the diagnosis of bacterial diarrhea were 94.2% and 22.8%. Salmonella were susceptible to ciprofloxacin, ceftriaxone and amoxicillin-clavulanate and showed low frequency of resistance to azithromycin. Campylobacter showed low frequency of resistance to azithromycin and high frequency of resistance to ciprofloxacin. Diarrheagenic Escherichia coli was highly susceptible to ciprofloxacin and amoxicillin-clavulanate. Contact with diarrheal patients was a risk factor for rotavirus [adjusted odds ratio (aOR): 11.7], norovirus (aOR: 7.5) and Campylobacter (aOR: 27.1) infections. Mother's education was positively associated with Salmonella infection (aOR: 2.1). Good hand hygiene was protective against rotavirus (aOR: 0.6), norovirus (aOR: 0.5) and Salmonella (aOR: 0.3) infections. Rotavirus, norovirus, nontyphoidal Salmonella and Campylobacter are significantly associated with diarrhea in Chinese children. Fecal leukocytes >5 per high-power field can predict bacterial diarrhea. Target prevention and appropriate treatment of diarrhea should consider the potential pathogen and resistance pattern.

Mechanisms of resistance against NITD-916, a direct inhibitor of Mycobacterium tuberculosis InhA

Isoniazid inhibits Mycobacterium tuberculosis InhA and is a key component of drug regimens that treat tuberculosis. However, the high rate of resistance against isoniazid is a contributing factor to the emergence of multi-drug resistance strains of M. tuberculosis. The 4-hydroxy-2-pyridine NITD-916 is a direct inhibitor of M. tuberculosis InhA that has comparable efficacy to isoniazid in mouse models of TB infection but a lower frequency of resistance. To characterize resistance mechanisms against NITD-916 we isolated resistant mutants in H37Rv (Euro-American lineage) and HN878 (East-Asian lineage) strains of M. tuberculosis. The resistance frequency was similar in both strains. Mutations were identified in residues within or near to the active of InhA or in the fabG1inhA promoter region. All mutants were resistant to NITD-916 but were not cross resistant to isoniazid, despite homology to SNPs identified in isoniazid resistant clinical isolates.

Phenotypic Characteristics and Genetic Diversity of Salmonella enterica Serotype Derby Isolated from Human Patients and Foods of Animal Origin.

Salmonella enterica serotype Derby is among the three most common serotypes of nontyphoidal Salmonella isolated from patients with diarrhea in China. In this study, 133 Salmonella Derby isolates from human patients (n = 74) and foods of animal origin (n = 59) in Shanghai, China, between September 2013 and December 2014, were selected to study its phenotypic characteristics and genetic diversity. The isolates were subjected to antimicrobial susceptibility testing, plasmid replicon typing, virulence profile determination, and molecular subtyping by pulsed-field gel electrophoresis (PFGE). Isolates were frequently resistant to tetracycline (87.22%), sulfisoxazole (74.44%), and streptomycin (62.41%), and a low frequency of resistance was found toward ofloxacin (3.01%), ceftazidime (2.26%), and cefepime (1.50%); in addition, 93 (69.92%) isolates were multidrug resistant. The most common plasmid incompatibility replicon types were the IncF family (FIA, 51.31%; FIC, 27.82%; and FIB, 21.80%) and IncP types (35.34%): these plasmid types may be associated with the spread of antibiotic resistance and virulence genes. All isolates were positive for the Salmonella pathogenicity island (SPI) gene avrA and the fimbrial gene bcfC from among the 10 virulence genes detected, and most of them carried ssaQ (99.25%), mgtC (97.74%), siiD (98.50%), sopB (97.74%), and sopE (96.99%). PFGE showed 68 patterns in nine main clusters at an 85% similarity threshold. Most of the isolates from different sources possessed the same fingerprints or molecular profiles in each cluster, which strongly suggests the possibility that foods of animal origin, especially pork, serve as an important source for human infection. Moreover, this diversity may suggest strains originating from multiple clones. Therefore, surveillance on this serotype should be strengthened to prevent transmission of Salmonella Derby from foods of animal origin, especially pork, to humans.

Antimicrobial susceptibility of bacterial isolates from patients presenting with ear, nose and throat (ENT) infections in the German community healthcare setting.

Empiric initial antibiotic therapy of bacterial infections is based primarily upon the susceptibility of the most common causative pathogens. The purpose of this study was to provide susceptibility data on six bacterial species known to cause ear, nose and throat (ENT) infections. A total of 1066 isolates collected during a nationwide laboratory-based surveillance study were analysed. All Streptococcus pyogenes isolates were penicillin (PEN)-susceptible, indicating that natural penicillins can still be recommended as the first-line treatment for group A streptococcal tonsillopharyngitis. Of the S. pneumoniae isolates, 92.9% were PEN-susceptible and of the Haemophilus influenzae isolates, 89.7% were amoxicillin-susceptible, retaining aminopenicillins as the first-line treatment for acute otitis media (AOM) and acute rhinosinusitis (ARS), in case antibiotic therapy is considered. In contrast, cefuroxime axetil seems less likely to be suitable for the treatment of AOM or ARS, as all Moraxella catarrhalis and >99% of the H. influenzae isolates were categorised as intermediate or resistant. The susceptibility rates of Pseudomonas aeruginosa were 97-100% for the drugs tested, except for the fluoroquinolones (87.6%). Overall, bacterial isolates from outpatients presenting with ENT infections showed low frequencies of resistance in Germany. However, given the emergence of multidrug resistance to standard antibiotics in Escherichia coli and other pathogens, inappropriate use of broad-spectrum antibiotics for the treatment of ENT infections has to be avoided.

Current and future antimicrobial resistance issues for the Australian pig industry

Antimicrobial use and antimicrobial resistance (AMR) in intensive pig production and its potential impacts to human and animal health are very much under the spotlight, both internationally, and within Australia. While the majority of AMR of medical importance is associated with the exclusive use of antimicrobials in humans, resistance in zoonotic foodborne pathogens such as Salmonella and Campylobacter, and livestock commensal bacteria such as Escherichia coli and Enterococcus spp., is under increased scrutiny. This is primarily due to the current reliance on many of the same drug classes as used in human medicine for treatment and control of bacterial diseases of livestock. Furthermore, the development of multidrug resistance in pathogens such as enterotoxigenic E. coli may drive off-label use of critically important drug classes such as 3rd-generation cephalosporins. This could lead to the emergence and amplification of resistance genes of potential public health significance in both pathogens and commensal bacteria. Livestock-associated and community-associated methicillin-resistant Staphylococcus aureus has also recently been detected in Australian pigs as a result of human-to-animal transmission and are a potential public health issue for in-contact piggery workers. Australia is in a unique position compared with many of its international trading partners due to its isolation, ban on importation of livestock and conservative approach to antimicrobial registration, including reservation of the fluoroquinolone class for use in humans and companion animals only. Cross-sectional AMR surveys of pathogens and commensals in healthy pigs have identified only low frequency of resistance to critically important drug classes. Nevertheless, resistance to critically important antimicrobials has emerged and careful antimicrobial stewardship is required to ensure that these low levels do not increase. In this report, we review AMR of significance to the Australian pig industry and identify potential prevention and control measures.

Diaryltriazenes as antibacterial agents against methicillin resistant Staphylococcus aureus (MRSA) and Mycobacterium smegmatis

Diaryltriazene derivatives were synthesized and evaluated for their antimicrobial properties. Initial experiments showed some of these compounds to have activity against both methicillin-resistant strains of Staphylococus aureus (MRSA) and Mycobacterium smegmatis, with MICs of 0.02 and 0.03 μg/mL respectively. Those compounds with potent anti-staphylococcal and anti-mycobacterial activity were not found to act as growth inhibitors of mammalian cell lines or yeast. Furthermore, we demonstrated that one of the most active anti-MRSA diaryltriazene derivatives was subject to very low frequencies of resistance at <10−9. Whole genome sequencing of resistant isolates identified mutations in the enzyme that lysylates phospholipids. This could result in the modification of phospholipid metabolism and consequently the characteristics of the staphylococcal cell membrane, ultimately modifying the sensitivity of these pathogens to triazene challenge. Our work has therefore extended the potential range of triazenes, which could yield novel antimicrobials with low levels of resistance.

Diarrheagenic Escherichia Coli (DEC) : Detection by Multiplex PCR and Antibiotic Susceptibility Pattern

Background : Diarrheagenic Escherichia coli (DEC) are among the most important causes of diarrheal diseases in children. O bjectives: This study aimed to detect the virulence loci associated with 4 categories of DEC among Egyptian children with and without diarrhea, and to get an idea about their antibiotic resistance pattern. Methodology: E coli were isolated from 104 diarrheal fecal samples and 104 non-diarrhea fecal samples. Multiplex PCR was performed to characterize target genes of DEC. Drug susceptibility patterns were determined by disc- diffusion method. Results: The target genes identifying DEC were detected in 21 diarrheal specimens (20.2%) including genes for ETEC (8 specimens, 7.7%); EPEC (6 specimens, 5.8%); EAEC (6 specimens, 5.8%) and one strain (1%) positive for genes of both ETEC and EPEC. DEC genes were detected in 3 out of 104 non-diarrheal specimens (2.9%); ETEC, 1% and EAEC, 2% with high significant difference between diarrheal and non-diarrheal groups. Antimicrobial susceptibility testing revealed high frequency of resistance of two groups to ampicillin, tetracycline and trimethoprim-sulphamethoxale (76.2% were multidrug resistant) ,low frequency of resistance to ampicillin-sulbactam, ceftazidim, ceftriaxone and azetronam and no resistance to cefepime, imipenem, amikacin or gentamicin. Conclusion : DEC is a major pathogen in childhood diarrhea in Egypt and the strains being susceptible to multiple antibiotics, although a large proportion (76.2%) of them were multi- drug resistant

Discovery and development of kibdelomycin, a new class of broad-spectrum antibiotics targeting the clinically proven bacterial type II topoisomerase

Kibdelomycin is a complex novel antibiotic, discovered by applying a highly sophisticated chemical-genetic Staphylococcus aureus Fitness Test (SaFT) approach, that inhibits the clinically established bacterial targets, gyrase and topoisomerase IV. It exhibits broad-spectrum antibacterial activity against aerobic bacteria including MRSA and Acinetobacter baumannii. It is slowly bactericidal and has a low frequency of resistance. In an anaerobic environment, it exhibits narrow-spectrum activity and inhibits the growth of gut bacteria Clostridium difficile (MIC 0.125μg/mL) without affecting the growth of commensal Gram-negative organisms particularly, Bacteroides sp. It is highly efficacious in the hamster model of C. difficile infection providing 100% protection at >6mg/kg and 80% protection at 1.56mg/kg by oral dosing without systemic exposure. X-ray co-crystal structures of kibdelomycin bound to GyrB and ParE showed a unique dual arm ‘U shaped’ multisite binding never encountered with any other gyrase inhibitors. Kibdelomycin is poised for preclinical development for C. difficile treatment, and most importantly, the co-crystal structures of kibdelomycin provide unique insight for structure-guided structure modification, which could lead to better broader-spectrum systemic antibiotic potentially covering many ESKAPE pathogens.

Development of near-isogenic lines with different alleles of Piz locus and analysis of their breeding effect under Yangdao 6 background

Rice blast is one of the most destructive diseases of rice. The most effective way of managing this disease is to develop resistant cultivars by introducing resistance genes into elite rice recipients. In this study, the near-isogenic lines (NILs) of six resistance alleles of the Piz locus (Pizt, Pi2, Pigm, Pi40, Pi9 and Piz) were constructed with Yangdao 6 as genetic background. Seedling inoculation tests showed that most of the NILs, namely NIL-Pi2, NIL-Pigm, NIL-Pi9, NIL-Pizt and NIL-Pi40, exhibited good resistance to blast with resistance frequencies (RFs) of over 82.50 %, execpt NIL-Piz which showed lower resistance with a RF of only 36.13 %. Furthermore, the improved-resistance NILs exhibited high similarity of their resistance spectra, with overlapping degrees of resistance spectrum (OD) of more than 75.83 %. However, the RF of panicle blast for all NILs decreased significantly compared with seedling blast in an artificial inoculation test. Although NIL-Pigm showed a higher panicle blast RF of 80 %, other NILs with outstanding performance in seedling blast resistance, namely NIL-Pizt, NIL-Pi2, NIL-Pi9 and NIL-Pi40, exhibited middle or low RFs of panicle blast with values from 56.67 to 33.30 %. Natural induction in a disease nursery showed a consistent trend in artificial inoculation results at seedling and heading stages. While NIL-Pigm was found to exhibit good resistance to leaf blast and panicle blast, NIL-Pi9 and NIL-Pizt were further demonstrated to show excellent resistance in Suichuan, Jiangxi province and Enshi, Hubei province, respectively, because of the race–region specificity. Agronomic traits of NILs were also investigated in order to evaluate the linkage drag effect of different alleles of the Piz locus. The resistance effects of the different alleles of the Piz locus under identical genetic background against seedling blast and panicle blast was first reported in this study, and the above results are expected to provide a theoretical support for the rational utilization of broad-spectrum resistance genes in breeding practice.

Frequencies of heavy metal resistance are associated with land cover type in the Upper Mississippi River

Taxonomic compositions of freshwater bacterial communities have been well-characterized via metagenomic and amplicon-based approaches, especially next-generation sequencing. However, functional diversity of these communities remains less well-studied. Various anthropogenic sources are known to impact the bacterial community composition in freshwater riverine systems and potentially alter functional diversity. In this study, high-throughput functional screening of large (~10,000 clones) fosmid libraries representing communities in the Upper Mississippi River revealed low frequencies of resistance to heavy metals in the following order: Mn2+>Cr3+>Zn2+>Cd2+>Hg2+. No resistance to Cu2+ was detected. Significant, but weak, correlations were observed between resistance frequencies of Cd and Cr with developed land cover (r2=0.08, P=0.016 and r=0.07, P=0.037, respectively). While discriminant function analyses further supported these associations, redundancy analysis further indicated associations with forested land cover and greater resistance to Hg and Zn. Nutrient and metal ion concentrations and abundances of bacterial orders were poorly correlated with heavy metal resistance, except for an association of Pseudomonadales abundance and resistance to Hg and Zn. Taken together, results of this study suggest that allochthonous bacteria contributed from specific land cover types influence the patterns of metal resistance throughout this river.

Molecular mechanisms of sulbactam antibacterial activity and resistance determinants in Acinetobacter baumannii.

Sulbactam is a class A β-lactamase inhibitor with intrinsic whole-cell activity against certain bacterial species, including Acinetobacter baumannii. The clinical use of sulbactam for A. baumannii infections is of interest due to increasing multidrug resistance in this pathogen. However, the molecular drivers of its antibacterial activity and resistance determinants have yet to be precisely defined. Here we show that the antibacterial activities of sulbactam vary widely across contemporary A. baumannii clinical isolates and are mediated through inhibition of the penicillin-binding proteins (PBPs) PBP1 and PBP3, with very low frequency of resistance; the rare pbp3 mutants with high levels of resistance to sulbactam are attenuated in fitness. These results support further investigation of the potential clinical utility of sulbactam.

In vitro antibacterial activity of AZD0914, a new spiropyrimidinetrione DNA gyrase/topoisomerase inhibitor with potent activity against Gram-positive, fastidious Gram-Negative, and atypical bacteria.

AZD0914 is a new spiropyrimidinetrione bacterial DNA gyrase/topoisomerase inhibitor with potent in vitro antibacterial activity against key Gram-positive (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumoniae, Streptococcus pyogenes, and Streptococcus agalactiae), fastidious Gram-negative (Haemophilus influenzae and Neisseria gonorrhoeae), atypical (Legionella pneumophila), and anaerobic (Clostridium difficile) bacterial species, including isolates with known resistance to fluoroquinolones. AZD0914 works via inhibition of DNA biosynthesis and accumulation of double-strand cleavages; this mechanism of inhibition differs from those of other marketed antibacterial compounds. AZD0914 stabilizes and arrests the cleaved covalent complex of gyrase with double-strand broken DNA under permissive conditions and thus blocks religation of the double-strand cleaved DNA to form fused circular DNA. Whereas this mechanism is similar to that seen with fluoroquinolones, it is mechanistically distinct. AZD0914 exhibited low frequencies of spontaneous resistance in S. aureus, and if mutants were obtained, the mutations mapped to gyrB. Additionally, no cross-resistance was observed for AZD0914 against recent bacterial clinical isolates demonstrating resistance to fluoroquinolones or other drug classes, including macrolides, β-lactams, glycopeptides, and oxazolidinones. AZD0914 was bactericidal in both minimum bactericidal concentration and in vitro time-kill studies. In in vitro checkerboard/synergy testing with 17 comparator antibacterials, only additivity/indifference was observed. The potent in vitro antibacterial activity (including activity against fluoroquinolone-resistant isolates), low frequency of resistance, lack of cross-resistance, and bactericidal activity of AZD0914 support its continued development.

Structures of kibdelomycin bound to Staphylococcus aureus GyrB and ParE showed a novel U-shaped binding mode.

Bacterial resistance to antibiotics continues to pose serious challenges as the discovery rate for new antibiotics fades. Kibdelomycin is one of the rare, novel, natural product antibiotics discovered recently that inhibits the bacterial DNA synthesis enzymes gyrase and topoisomerase IV. It is a broad-spectrum, Gram-positive antibiotic without cross-resistance to known gyrase inhibitors, including clinically effective quinolones. To understand its mechanism of action, binding mode, and lack of cross-resistance, we have co-crystallized kibdelomycin and novobiocin with the N-terminal domains of Staphylococcus aureus gyrase B (24 kDa) and topo IV (ParE, 24 and 43 kDa). Kibdelomycin shows a unique "dual-arm", U-shaped binding mode in both crystal structures. The pyrrolamide moiety in the lower part of kibdelomycin penetrates deeply into the ATP-binding site pocket, whereas the isopropyl-tetramic acid and sugar moiety of the upper part thoroughly engage in polar interactions with a surface patch of the protein. The isoproramic acid (1,3-dioxopyrrolidine) and a tetrahydropyran acetate group (Sugar A) make polar contact with a surface area consisting of helix α4 and the flexible loop connecting helices α3 and α4. The two arms are connected together by a rigid decalin linker that makes van del Waals contacts with the protein backbone. This "dual-arm", U-shaped, multicontact binding mode of kibdelomycin is unique and distinctively different from binding modes of other known gyrase inhibitors (e.g., coumarins and quinolones), which explains its lack of cross-resistance and low frequency of resistance. The crystal structures reported in this paper should enable design and discovery of analogues with better properties and antibacterial spectrum.

Phosphine Resistance in &lt;I&gt;Tribolium castaneum&lt;/I&gt; and &lt;I&gt;Rhyzopertha dominica&lt;/I&gt; From Stored Wheat in Oklahoma

Phosphine gas, or hydrogen phosphide (PH3), is the most common insecticide applied to durable stored products worldwide and is routinely used in the United States for treatment of bulk-stored cereal grains and other durable stored products. Research from the late 1980s revealed low frequencies of resistance to various residual grain protectant insecticides and to phosphine in grain insect species collected in Oklahoma. The present work, which used the same previously established discriminating dose bioassays for phosphine toxicity as in the earlier study, evaluated adults of nine different populations of red flour beetle, Tribolium castaneum (Herbst), and five populations of lesser grain borer, Rhyzopertha dominica (F.) collected from different geographic locations in Oklahoma. One additional population for each species was a laboratory susceptible strain. Discriminating dose assays determined eight out of the nine T. castaneum populations, and all five populations of R. dominica, contained phosphine-resistant individuals, and highest resistance frequencies were 94 and 98%, respectively. Dose-response bioassays and logit analyses determined that LC99 values were approximately 3 ppm for susceptible and 377 ppm for resistant T. castaneum, and approximately 2 ppm for susceptible and 3,430 ppm for resistant R. dominica. The most resistant T. castaneum population was 119-fold more resistant than the susceptible strain and the most resistant R. dominica population was over 1,500-fold more resistant. Results suggest a substantial increase in phosphine resistance in these major stored-wheat pests in the past 21 yr, and these levels of resistance to phosphine approach those reported for other stored-grain pest species in other countries.

Antibiotic resistance and restriction endonucleases in fecal enterococci of chamois (Rupicapra rupicapra Linnaeus, 1758)

Two hundred eighty-four isolates of enterococci from feces of wild living chamois from alpine environments were tested for sensitivity to three antibiotics. Low frequency of resistance was observed in studied enterococcal populations (about 5% for tetracycline and erythromycin and 0% for ampicillin). In six animals, the population of enterococci lacked any detectable resistance. Our data indicated that enterococcal population in feces of the majority of studied animals did not encounter mobile genetic elements encoding antibiotic resistance probably due to spatial separation and/or due to low exposure to the antibiotics. Based on resistance profiles observed, three populations were analyzed for the presence of restriction endonucleases. The restriction enzymes from two isolates-31K and 1K-were further purified and characterized. Restriction endonuclease Efa1KI recognizes CCWGG sequence and is an isoschizomer of BstNI. Endonuclease Efc31KI, a BsmAI isoschizomer, recognizes the sequence GTCTC and it is a first restriction endonuclease identified in Enterococcus faecium. Our data indicate that restriction-modification (R-M) systems do not represent an efficient barrier for antibiotic resistance spreading; enterococcal populations colonized by antibiotics resistance genes were also colonized by the R-M systems.

The oxazolidinones: past, present, and future

The success of linezolid stimulated significant efforts to discover new agents in the oxazolidinone class. Over a dozen oxazolidinones have reached the clinic, but many were discontinued due to lack of differentiated potency, inadequate pharmacokinetics, and safety risks that included myelosuppression. Four oxazolidinones are currently undergoing clinical evaluation. The Trius Therapeutics compound tedizolid phosphate (formerly known as torezolid phosphate, TR-701, DA-7218), the most advanced, is in phase 3 clinical trials for acute bacterial skin and skin structure infections. Rib-X completed two phase 2 studies for radezolid (Rx-01_667, RX-1741) in uncomplicated skin and skin structure infections and community-acquired pneumonia. Pfizer and AstraZeneca have each identified antitubercular compounds that have completed phase 1 studies: sutezolid (PNU-100480, PF-02341272) and AZD5847 (AZD2563), respectively. The oxazolidinones share a relatively low frequency of resistance largely due to the requirement of mutations in 23S ribosomal RNA genes. However, maintaining potency against strains carrying the mobile cfr gene poses a challenge for the oxazolidinone class, as well as other 50S ribosome inhibitors that target the peptidyl transferase center.

Pilot Study of Antimicrobial-Resistant Escherichia coli in Herring Gulls (Larus argentatus) and Wastewater in the Northeastern United States

Wildlife may be an important reservoir of antibiotic-resistant bacteria and resistance genes. In this pilot study, the prevalence and patterns of antimicrobial resistance in Escherichia coli cultured from wild herring gull (Larus argentatus) feces and human wastewater at Cape Cod, Massachusetts, USA, was compared. Antimicrobial susceptibility was tested using Kirby-Bauer disk diffusion with seven antimicrobial agents. A high proportion of antimicrobial agent-resistant E. coli isolates (59.2%) were detected in wastewater samples compared with a lower prevalence of 17.5% in gull feces. In addition, there was a large proportion of isolates with intermediate susceptibility (93.0%) in gull feces. Although similar resistance patterns and shared resistance genes suggest possible wastewater contamination of the local environment, the relatively low frequency of resistance and high prevalence of intermediate susceptibility detected in E. coli cultured from gull feces depict a complex model of antimicrobial resistance among E. coli strains of wildlife origin.

Genetic Diversity and Population Structure in Aegilops sharonensis

Sharon goatgrass (Aegilops sharonensis Eig) is a useful source of genes for wheat (Triticum aestivum L.) improvement, but little is known about its genetic diversity in Israel. We analyzed genetic diversity and population structure of 106 Ae. sharonensis accessions. Twenty‐one microsatellites revealed a high level of diversity that was comparable to Triticeae species with larger geographic distribution. The genetic structure of Ae. sharonensis is characterized by high diversity within location and by a short distance (15 km) similarity between populations. Accessions from the Sharon Plain exhibited the highest level of diversity. Because the Sharon Plain is located in the center of geographic distribution of the species and includes the three most genetically diverse sites, it may be the center of diversity of Ae. sharonensis A wide range of variation was observed in the 11 morphological traits evaluated, and the regional effect on the phenotypic variability was higher than the one observed for genetic diversity. Considerable variability was found in response to leaf rust and stem rust among regions, as a very low frequency of resistance was found for both diseases in the northern regions of Israel. The diversity pattern found in populations of Ae. sharonensis indicate that, to optimize the capture of the greatest diversity, collection and utilization programs should include multiple individuals from as many sites as possible.