Electrical characterization of composition modulated In(1-x)Sb(x) nanowire field effect transistors by scanning gate microscopy.

In this work high quality crystalline In(1_x)Sb(x) nanowires (NWs) are synthesized via a template-based electrochemistry method. Energy dispersive spectroscopy studies show that composition modulated In(1-x)Sb(x) (x approximately 0.5 or 0.7) nanowires can be attained by selectively controlling the deposition potential during growth. Single In(1-x)Sb(x) nanowire field effect transistors (NW-FETs) are fabricated to study the electrical properties of as-grown NWs. Using scanning gate microscopy (SGM) as a local gate the I(ds)-V(ds) characteristics of the fabricated devices are modulated as a function of the applied gate voltage. Electrical transport measurements show n-type semiconducting behavior for the In0.5Sb0.5 NW-FET, while a p-type behavior is observed for the In0.3Sb0.7 NW-FET device. The ability to grow composition modulated In(1-x)Sb(x) NWs can provide new opportunities for utilizing InSb NWs as building blocks for low-power and high speed nanoscale electronics.

DNA barcoding Indian freshwater fishes.

DNA barcoding is a promising technique for species identification using a short mitochondrial DNA sequence of cytochrome c oxidase I (COI) gene. In the present study, DNA barcodes were generated from 72 species of freshwater fish covering the Orders Cypriniformes, Siluriformes, Perciformes, Synbranchiformes, and Osteoglossiformes representing 50 genera and 19 families. All the samples were collected from diverse sites except the species endemic to a particular location. Species were represented by multiple specimens in the great majority of the barcoded species. A total of 284 COI sequences were generated. After amplification and sequencing of 700 base pair fragment of COI, primers were trimmed which invariably generated a 655 base pair barcode sequence. The average Kimura two-parameter (K2P) distances within-species, genera, families, and orders were 0.40%, 9.60%, 13.10%, and 17.16%, respectively. DNA barcode discriminated congeneric species without any confusion. The study strongly validated the efficiency of COI as an ideal marker for DNA barcoding of Indian freshwater fishes.

Revised estimates of enterococcal chromosomal sizes.

We previously reported that the chromosomal sizes of four strains of enterococci ranged from 2,045 to 2,761 kb. Extensive analysis and mapping subsequently confirmed the size of Enterococcus faecalis strain OG1 as 2,825 kb (prior size estimate range, 2,750-2,761 kb) (Murray et al., J. Bacteriol. 175, 5216, 1993). However, using variable conditions of electrophoresis and additional digestions, revised size estimates for the other strains are 2,852-3,093 kb for E. faecalis strain JH2-2 (prior range, 2,008-2,135 kb), 2,910-3,065 kb for E. faecalis strain HH67 (prior range, 2,170-2,288 kb), and 2,334-2,558 for E. faecium strain GE-1 (prior range, 2,045-2,155 kb). The earlier underestimations of the chromosomal sizes were due to the inconsistent presence of a large fragment, likely caused by shearing of the DNA during handling, causing it to be considered a partial digestion product, and failure to resolve multiple fragments of the same approximate size.

Determination of the chromosomal size of three different strains of Enterococcus faecalis and one strain of Enterococcus faecium.

Pulsed-field gel electrophoresis was used to determine the chromosomal size of three different strains of Enterococcus faecalis and one strain of Enterococcus faecium. The size determinations of OG1X, a strain of E. faecalis widely used in many laboratories for genetic studies, using Sma I, Not I, and Sfi I alone or in combination, ranged from 2,750 to 2,761 kb. Using the same enzymes as with OG1X, the size of HH-67, a plasmid-free clinical isolate of E. faecalis, was determined to be 2,170-2,288 kb and the size of JH2-2, an E. faecalis recipient strain, ranged from 2,008 to 2,135 kb. The size range generated for GE-1, a plasmid-free E. faecium strain, with the use of Sma I, Not I, and Apa I was 2,045-2,155 kb. Although OG1X differed in size from the other three enterococci, each individual enterococcal strain generated reproducible results in different experiments. However, for both E. faecalis OG1X and E. faecium GE-1, one of the enzymes used generated a considerably smaller molecular size than that generated by the other two enzymes. The discrepancy was due to visually undiscernible comigrating fragments, and serves to point out a potential source of error if fewer than two enzymes are used to size a genome. The size discrepancies were resolved by digesting individual fragments with a second enzyme. The molecular sizes of these enterococcal strains are larger than that recently reported for Campylobacter, smaller than that of Escherichia coli and Pseudomonas aeruginosa, and similar (OG1X) or smaller (JH2-2, HH67, and GE-1) than the 2,819-kb reported for Streptococcus mutans.

Streptogramin resistance and shared pulsed-field gel electrophoresis patterns in vanA-containing Enterococcus faecium and Enterococcus hirae isolated from humans and animals in Spain.

The present study was performed to determine if any of the 45 vanA-containing Enterococcus faecium or 18 vanA-containing E. hirae strains were shared by chickens (32 E. faecium/l7 E. hirae) and humans (13 E. faecium/1 E. hirae) using pulsed field gel electrophoresis (PFGE) and to study quinupristin-dalfopristin (Q-D) resistance. Seven of the 45 E. faecium isolates (from 2 outpatients and from 5 poultry products) were resistant to Q-D (MIC > or = 16 microg/ml); one strain was shown to have satA by PCR and sequencing and, in the other six isolates, the recently described satG gene was demonstrated. Six different PFGE patterns were detected among the 7 Q-D E. faecium-resistant isolates. None of the E. hirae isolates showed Q-D resistance. Among the 45 vanA -containing E. faecium strains, 25 unrelated clones were found by PFGE with highly diverse patterns and an indistinguishable PFGE pattern was observed in vanA-containing E. faecium strains from two humans and two poultry products. A single PFGE pattern was detected in 17 of 18 vanA-containing E. hirae isolates, obtained from one human and 16 chicken samples. Based on the presence of indistinguishable PFGE patterns among VR E. faecium and E. hirae from humans and chickens, we conclude that horizontal transfer of these strains could occur between both groups.

In vitro activity of 19 antimicrobial agents against enterococci from healthy subjects and hospitalized patients and use of an ace gene probe from Enterococcus faecalis for species identification.

We tested 165 enterococcal isolates, biased toward vancomycin resistant (VR) isolates, collected during recent years from fecal samples of healthy subjects and clinical specimens of hospitalized patients (mostly from United States and some from Europe) for susceptibility to 19 antimicrobials. Nosocomial isolates, whether VR or not, were more often highly resistant to aminoglycosides and clindamycin than fecal isolates from healthy community volunteers and more often resistant to erythromycin, chloramphenicol, trimethoprim, levofloxacin and, for E. faecium, ampicillin (93 vs. 0%). Resistance rates were similar between nosocomial and community-fecal isolates for minocycline, rifampin and quinupristin-dalfopristin (Q-D). None of the 165 enterococci tested hybridized with aph(2'')-Ic and aph(2'')-Id probes for recently described gentamicin resistance genes and 37 of the 39 isolates with high level resistance (HLR) to gentamicin hybridized with an intragenic aac(6')-aph(2'') probe. Of the two newer drugs tested, daptomycin MIC90s were 0.25 microg/mL for E. faecalis and 1 microg/mL for E. faecium, regardless of their vancomycin resistance level or source. For Q-D, none of 28 E. faecium from community based healthy subjects in the USA and 7 of 66 E. faecium from hospitalized patients in the United States were resistant. Among these 7 Q-Dr United States isolates and 7 Q-Dr isolates from Europe (MICs of Q-D of 4-8 microg/mL), none hybridized with vat(D) (formerly satA) and vat(E) (formerly satG) DNA probes, indicating the involvement of other mechanism/s of resistance in these isolates. We also demonstrated that an intragenic probe of the gene ace from E. faecalis showed specific hybridizations to all E. faecalis isolates, suggesting the usefulness of this gene for identification of this species.

In vitro activity of moxifloxacin, a new 8-methoxyquinolone, against gram-positive bacteria.

The in vitro activity of moxifloxacin, formerly BAY 12-8039, against gram-positive bacteria was tested by the agar dilution method. A total of 189 isolates that included Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, streptococci, rhodococci, leuconostocs, pediococci, lactobacilli, and diphtheroids were tested. Moxifloxacin showed greater potency than ciprofloxacin against S. aureus, streptococci, and enterococci, having Minimal Inhibitory Concentrations (MICs) lower than those of ciprofloxacin by 2- to 64-fold. This improved activity was most prominent for S. aureus. Moxifloxacin was active against Leuconostoc and Rhodococcus species. Time-kill studies using moxifloxacin at a concentration of 3 micrograms/mL against one isolate each of methicillin-resistant S. aureus (MSSA) (MIC, 0.031 microgram/mL), MRSA (MIC, 1 microgram/mL), two isolates of E. faecalis (MICs, 0.25 and 2 micrograms/mL), and two isolates of vancomycin-resistant E. faecium (MICs, 0.25 and 2 micrograms/mL) revealed an average decrease in colony forming unit (CFU) by 3.8, 0.4, 4.0, 2.0, 4.2, and 1.8 log10 CFU/mL at 24 h, respectively. Moxifloxacin is a new 8-methoxyquinolone with improved in vitro activity against gram-positive bacteria. Further studies of the in vivo activity of this compound appear warranted.

Susceptibility of beta-lactamase-producing enterococci to piperacillin with tazobactam.

The in vitro activity of piperacillin with and without tazobactam was evaluated against different inocula of 12 clinical isolates of beta-lactamase-producing Enterococcus faecalis obtained from different geographic areas. Minimum inhibitory concentrations (MICs) of piperacillin alone at approximately 10(3) colony-forming units (CFU)/spot ranged from 4 to 8 and from 4 to 8 micrograms/ml with piperacillin plus tazobactam. When approximately 10(7) CFU/spot was used, MICs increased to a range of 128-1024 micrograms/ml piperacillin. This inoculum effect was reversed by the addition of tazobactam to piperacillin at a fixed concentration of 1 microgram/ml or at a ratio of 8 : 1 (piperacillin relative to tazobactam) with an MIC90 of 16/2 micrograms/ml for the combination drug. In time-kill studies, four beta-lactamase-producing (Bla+) isolates were tested and demonstrated a decrease of > or = 2 log10 with 8 or 16 micrograms/ml of piperacillin in combination with 4 micrograms of tazobactam, but not with piperacillin alone. A non-beta-lactamase-producing isolate was equally inhibited by piperacillin alone and piperacillin plus tazobactam. Against a Bla+ isolate, the combination of piperacillin with tazobactam with streptomycin resulted in a synergistic effect relative to that of piperacillin with tazobactam; piperacillin plus streptomycin did not show synergism. Piperacillin in combination with tazobactam is active against enterococci that produce beta-lactamase and, in combination with an appropriate aminoglycoside, could be a viable choice for therapy of enterococci that do not have high-level resistance to all aminoglycosides.

In vivo testing of an Enterococcus faecalis efaA mutant and use of efaA homologs for species identification.

Disruption of the previously described efaA (from Enterococcus faecalis antigen A) gene was generated in E. faecalis strain OG1RF and loss of an 37-kDa immunoreactive band from the mutant was demonstrated in Western blots. In a mouse peritonitis model, mice infected with the efaAfs (fs=from Enterococcus faecalis) mutant showed more prolonged survival than mice infected with the parent strain OG1RF. These results suggest that efaAfs encodes a function important for infection of mice by enterococci. An efaA-like gene was also identified in E. faecium DNA libraries and its deduced amino acid sequence showed 73% similarity to EfaA of E. faecalis and 42-63% similarities to a group of streptococcal virulence and adhesion associated proteins that are components of ATP-binding cassette transport systems. Intragenic probes representing efaAfs, recAfs, efaAfm (fm=from E. faecium) and gyrAfm were tested for their ability to identify E. faecalis and E. faecium using colony lysates of 133 enterococci and one Streptococcus sp. Probes of E. faecium and E. faecalis origin hybridized to all isolates of E. faecium and E. faecalis, respectively, regardless of their clinical source but not to any of 29 other enterococci. These results suggest that the use of gene probes may prove helpful in identification of isolates of E. faecium and E. faecalis.

Specific antibody promotes opsonization and PMN-mediated killing of phagocytosis-resistant Enterococcus faecium.

Many clinical isolates of Enterococcus faecium are resistant to neutrophil (PMN)-mediated phagocytosis and killing in the presence of normal human serum. We have now examined the ability of specific polyclonal rabbit antibodies to promote opsonization and killing of phagocytosis-resistant E. faecium. Immune rabbit serum generated against formalin-killed E. faecium TX0016, a phagocytosis-resistant strain, markedly promoted binding of TX0016 organisms to PMNs and PMN-mediated killing. These effects were dramatically reduced by (a) adsorption of immune serum with E. faecium TX0016, but not by adsorption with a strain of E. faecium susceptible to phagocytosis, and (b) incubation of immune serum with carbohydrate purified from TX0016, but not by incubation with a surface protein extract from TX0016. IgG purified from immune serum was unable by itself to promote bacterial binding to PMNs. However, specific IgG was able to promote binding to PMNs and PMN-mediated killing in the presence of normal human serum as a complement source, as were F(ab')(2) and Fab fragments produced from it, and the alternative pathway of complement was sufficient to promote IgG- and F(ab')(2)-mediated opsonization. PMN complement receptor type 3, but not complement receptor type 1, was involved in bacterial binding to PMNs induced by the combination of F(ab')(2) fragments and normal human serum. These results suggest that opsonization by antibodies potentially directed against bacterial carbohydrate, in conjunction with complement activation, has an important role in the host defense against phagocytosis-resistant E. faecium.

Vancomycin resistance factor of Lactobacillus rhamnosus GG in relation to enterococcal vancomycin resistance (van) genes.

Lactobacillus rhamnosus GG (ATCC 53103) is a probiotic strain used in fermented dairy products in many countries and is also used as a food supplement in the form of freeze-dried powder. The relationship of the vancomycin resistance factor in L. rhamnosus GG and the vancomycin resistance (van) genes of Enterococcus faecalis and E. faecium were studied using polymerase chain reaction (PCR), Southern hybridization and conjugation methods. Our results show that the vancomycin resistance determinant in L. rhamnosus GG is not closely related to enterococcal van genes, since no PCR product was amplified in L. rhamnosus GG with any of the three sets of vanA primers used, and enterococcal vanA, vanB, vnH, vanX, vanZ, vanY, vanS and vanR genes did not hybridize with DNA of L. rhamnosus GG. This strain does not contain plasmids and transfer of chromosomal vancomycin resistance determinant from L. rhamnosus GG to enterococcal species was not detected. Our results are in accordance with previous findings of intrinsically vancomycin-resistant lactic acid bacteria.

Vancomycin-resistant enterococci isolated from animals and food.

One hundred and one chicken products, boiled ham and turkey cold meat were acquired from 18 different supermarkets in Spain during October 1997 to June 1998 and were analyzed for vancomycin-resistant enterococci (VRE). In the same way, 50 intestinal chicken samples from a slaughterhouse were also studied. VRE were detected in 25 of 92 samples of food of chicken origin (27.2%), but no VRE were found in cooked pork or turkey products. VRE were also detected in 8 of 50 intestinal chicken samples from the slaughterhouse (16%). VRE were identified as Enterococcus durans (n = 11), Enterococcus faecalis (n = 10), Enterococcus faecium (n = 10) and Enterococcus hirae (n = 2). All these strains were characterized as belonging to the vanA genotype by polymerase chain reaction. Ampicillin, quinupristin/dalfopristin and high level aminoglycoside resistance were frequently found among these strains. Heterogeneity was observed in susceptibility patterns among VRE strains, even in those of the same species. The high rate of colonization of chicken products by vanA containing enterococci detected 6 months to 1 year after the banning of avoparcin as a growth promoter, supports other studies suggesting that the food chain could be a source of VRE colonization in humans and thus a source of VRE infections.

Application of molecular techniques to the study of nosocomial infections caused by enterococci.

Enterococci are components of the normal bowel flora of humans and other animals, and have traditionally been considered to be of relatively low virulence in healthy individuals. However, they are increasingly important nosocomial pathogens and have been cited as the leading organism isolated from hospital-acquired infections, and the third leading cause of nosocomial bacteremia in the United States in a recent National Nosocomial Infection Surveillance (NNIS) system report of the Centers for Disease Control (1). The increase in enterococcal infections has been associated with the emergence of resistance to multiple antibiotics, in particular resistance to B-lactams, high-level aminoglycoside resistance, and resistance to glycopeptides. Concern that antibiotic resistance will continue to spread and will increasingly render conventional antimicrobial chemotherapy inadequate for serious enterococcal infections has stimulated interest in methods to improve the diagnosis and epi-demiologic investigation of infections caused by enterococci.

Susceptibility status of two species of Japanese encephalitis vectors to insecticides in the Thar desert, district Bikaner (Rajasthan).

Susceptibility tests were conducted on the adults of two species of Japanese encephalitis vectors viz., Culex pseudovishnui and C. tritaeniorhynchus against diagnostic doses of DDT, dieldrin, malathion, fenitrothion, propoxur and permethrin at different exposure duration. C. pseudovishnui was found susceptible to permethrin and resistant to dieldrin and propoxur while C. tritaeniorhynchus was found susceptible to permethrin and resistant to DDT, dieldrin, fenitrothion and propoxur. A verification, however, was required with other insecticides for both the species.

Insecticide susceptibility of Phlebotomus papatasi to organochlorine, organophosphate & carbamate compounds in some arid areas of western Rajasthan.

Insecticide susceptibility status of P. papatasi to organochlorine, organophosphate and carbamate compounds has been estimated in Pali and Barmer districts of Rajasthan. Tests revealed that this species was resistant to DDT but susceptible to dieldrin, malathion, fenitrothion and propoxur. Efficacies to these compounds at 50 and 95 per cent levels have been estimated by probit-analysis. The LC50 values for both DDT and dieldrin, were found much lower than those reported from other parts of the country. Heterogeneity of the response was found highly significant [X2 = 43.8(3)] in case of propoxur only.

Current status of Anopheles stephensi response to various insecticides in some areas of the Thar desert.

Investigations on the current response of A. stephensi. to six insecticides viz. DDT, dieldrin, malathion, fenitrothion, propoxur and permethrin, were carried out in 3 districts i.e. Barmer, Jodhpur and Pali, of the Thar desert. The species was found resistant to DDT and dieldrin, partially resistant to malathion and susceptible to fenitrothion, propoxur and permethrin. Dieldrin and malathion resistance has been detected for the first time in the Thar desert. Lethal concentrations (LC50 & LC95) of DDT and dieldrin and lethal exposure times (LT50 & LT95) of malathion, fenitrothion, propoxur and permethrin have been determined. In some areas, the differences in LC50 and LT50 values of tested insecticides, except fenitrothion, were found statistically significant. Chi-square and regression tests have revealed the homogeniety and linear trend respectively in the response of A. stephensi to insecticides. The findings of the study indicate that organochlorine compounds can be used alternately in the spray operations.

Characteristics of a local virulent strain of Newcastle disease virus.

A local virulent strain, VLT, of Newcastle disease virus formed 3- to 4-mm plaques on monolayers of primary chicken embryo cultures on the 4th day after inoculation. It agglutinated chicken and human 0 erythrocytes. Its hemagglutinin was stable at 56 C when compared with those of Komarov (K) and F vaccinal strains of the same virus. The viral titer of infected allantoic fluid dropped from 10(8.1) plaque-forming units to 10(1.0) plaque-forming units/ml within 2 hours when incubated at 56 C. The strain was ether-sensitive; it adsorbed readily on monolayers of chicken embryo cells and did not diffuse through agar. Its intracerebral pathogenicity index, chicken dose LD50, and embryo mean death time (hours) were 1.8, 9.0, and 48, respectively. Two virulent strains isolated in 1974 and 1975 were found to be identical to the VLT strain in terms of certain physical and biological properties. On the basis of plaque morphologic characteristics, hemagglutination spectrum, and hemagglutinin inactivation at 56 C, it was possible to identify readily the field isolate when it was compared with vaccinal strains (K and F) commonly used in Lebanon.

Viral proliferation patterns of a velogenic (VLT), a mesogenic (Komarov), and a lentogenic (F) strain of Newcastle disease virus.

A highly pathogenic and two avirulent vaccine strains of Newcastle disease virus were investigated quantitatively for their proliferation in various tissues of experimentally infected SPF chickens. Virulent strain VLT multiplied extensively in all the tested tissues whereas a mesogenic strain (K) was not detected in the brain during the period of observations. A lentogenic strain (F) was only detected in moderate quantities in trachea. The development of antibodies seemed to correlate with the disappearance of the two avirulent strains from the tissues.

An unusual cause of gastric outlet obstruction: a case report.

We report the case of a 49-year-old male who presented with pain in the upper abdomen and loss of appetite of 1 month's duration. A diagnosis of an unusual amoebic liver abscess was made. The patient recovered well after aspiration of 600 ml of anchovy sauce pus and treatment with metronidazole.

Susceptibility status of Phlebotomus papatasi and Sergentomyia punjabaensis (Diptera:Psychodidae) to some insecticides in district Bikaner (Rajasthan).

Susceptibility tests were carried out on the females of two species of sandflies viz. Phlebotomus papatasi and Sergentomyia punjabaensis against six insecticides viz. DDT, dieldrin (organochlorines); malathion, fenitrothion (organophosphates); propoxur (carbamate) and permethrin (synthetic pyrethroid) in district Bikaner, Rajasthan. A concentration and time dependent effect was observed with insecticides for both the species. P. papatasi was found resistant to DDT, dieldrin and propoxur while susceptible to malathion, fenitrothion and permethrin. However, S. punjabensis was found susceptible to all the insecticides tested. LC50 and LT50 value estimated for DDT and dieldrin for P. papatasi and S. punjabaensis were found to be 2.2 and 0.3% x 1 hr and 1.45 and 0.032% x 15 min. respectively.