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Sample records for resistant protein isolated

  1. Differentially expressed proteins in fluconazole-susceptible and fluconazole-resistant isolates of Candida glabrata.

    Science.gov (United States)

    Shen, Yinzhong; Zhang, Lijun; Jia, Xiaofang; Zhang, Yongxin; Lu, Hongzhou

    2015-06-01

    The current study aimed to identify the differences presented in the proteome of fluconazole-susceptible isolates of Candida glabrata compared to those with fluconazole-resistant ones. Two-dimensional differential gel electrophoresis was applied to identify proteins that were differentially expressed in fluconazole-susceptible and fluconazole-resistant isolates of C. glabrata. Eight proteins including aspartyl-tRNA synthetase, translation elongation factor 3, 3-phosphoglycerate kinase, ribosomal protein L5, coproporphyrinogen III oxidase, pyruvate kinase, G-beta like protein, and F1F0-ATPase alpha subunit were found to be more abundantly represented, while four proteins including vitamin B12-(cobalamin)-independent isozyme of methionine synthase, microtubule-associated protein, adenylosuccinate synthetase, and aldose reductase were found to be less abundantly represented in fluconazole-resistant strains versus those with fluconazole-susceptible ones. These differentially expressed proteins were primarily associated with energy metabolism, stress response, and macromolecule synthesis. Proteins associated with energy metabolism, stress response, and macromolecule synthesis may play a role in the development of fluconazole resistance in the clinical isolates of C. glabrata. Multiple different mechanisms are involved in the development of fluconazole resistance in C. glabrata. These findings provide a scientific basis for discovering new genes and mechanisms associated with fluconazole resistance in C. glabrata.

  2. Total Protein Profile and Drug Resistance in Candida albicans Isolated from Clinical Samples

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    Kamal Uddin Zaidi

    2016-01-01

    Full Text Available This study was done to assess the antifungal susceptibility of clinical isolates of Candida albicans and to evaluate its total protein profile based on morphological difference on drug resistance. Hundred and twenty clinical isolates of C. albicans from various clinical specimens were tested for susceptibility against four antifungal agents, namely, fluconazole, itraconazole, amphotericin B, and ketoconazole. A significant increase of drug resistance in clinical isolates of C. albicans was observed. The study showed 50% fluconazole and itraconazole resistance at 32 μg mL−1 with a MIC50 and MIC90 values at 34 and 47 and 36 and 49 μg mL−1, respectively. All isolates were sensitive to amphotericin B and ketoconazole. The SDS-PAGE protein profile showed a prevalent band of ~52.5 kDa, indicating overexpression of gene in 72% strains with fluconazole resistance. Since the opportunistic infections of Candida spp. are increasing along with drug resistance, the total protein profile will help in understanding the evolutionary changes in drug resistance and also to characterize them.

  3. [Evaluation of penicillin-binding protein genotypes in penicillin susceptible and resistant Streptococcus pneumoniae isolates].

    Science.gov (United States)

    Aslan, Gönül; Tezcan, Seda; Delialioğlu, Nuran; Aydın, Fatma Esin; Kuyucu, Necdet; Emekdaş, Gürol

    2012-04-01

    Penicillin-binding proteins (PBPs) are the natural targets of beta-lactam antibiotics and mutations in pbp1a, pbp2b, and pbp2x genes, which encode PBPs, are responsible for resistance to beta-lactams in Streptococcus pneumoniae. In the present study, we intended to determine how often the common mutation patterns occurred within the pbp1a, pbp2b, and pbp2x PBP gene regions and evaluate the PBP genotype mutations which were associated with penicillin resistance in several penicillin-susceptible and - resistant S.pneumoniae isolates in Mersin, Turkey. A total of 62 S.pneumoniae strains isolated from different clinical specimens (32 nasopharyngeal swab, 16 sputum, 3 blood, 3 wound, 2 cerebrospinal fluids and one of each urine, abscess, bronchoalveolar lavage, conjunctival swab, tracheal aspirate, middle ear effusion) were included in the study. Penicillin susceptibilities of the isolates were searched by disc diffusion and E-test methods, and 23 of them were identified as susceptible, 31 were intermediate susceptible, and eight were resistant to penicillin. A rapid DNA extraction procedure was performed for the isolation of nucleic acids from the strains. Distribution of PBP gene mutations in pbp1a, pbp2b, and pbp2x gene regions related to penicillin resistance was determined by using a wild-type specific polymerase chain reaction (PCR) based technique. PBP gene alterations of those isolates were also evaluated in relation to penicillin susceptibility and resistance patterns. Twenty two (95.7%) of 23 penicillin-susceptible S.pneumoniae isolates exhibited no mutation in the three PBP genes (pbp1a, pbp2x, and pbp 2b), while 1 (4.3%) of these harbored mutations in all of the three PBP genes. The penicillin-intermediate susceptible S.pneumoniae isolates exhibited various combinations of mutations. One (3.2%) of 31 penicillin-intermediate susceptible isolates exhibited no mutation in the three PBP genes, while 22 (71%) of them yielded mutations in all of the three PBP

  4. Isolation and Identification of Alicyclobacillus with High Dipicolinic Acid and Heat Resistant Proteins from Mango Juice

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    Hamid Reza Akhbariyoon

    2016-10-01

    Full Text Available Background and Objectives: Microbial spoilage of juices and industries related with Alicyclobacillus are considerable international issues. This spore-forming bacterium causes changes in juices odor and taste. The isolation and identification of Alicyclobacillus contamination in juice producing and packaging industries has an essential role in the prevention and control of this type of spoilage bacterium in HACCP (Hazard analysis and critical control points manner.Materials and Methods: A thermo-acidophilic, non-pathogenic and sporeforming bacterium was isolated from mango juice. Preliminary identification of the isolates was based on morphological, biochemical and physiological properties. Identification at species level was made by PCR amplification. The influence of temperature in the range of 25-65°C in the growth of bacterium and in the range of 80-120°C in spore-resistant and heat resistant proteins was investigated and compared with other spore producing bacteria.Results and Conclusion: Phylogenetic analysis of the 16S rRNA gene sequencing indicated that the isolated strain constituted a distinct lineage in the Alicyclobacillus cluster and submitted to NCBI with access number Alicyclobacillus HRM-5 KM983424.1. The spores resisted 110°C for 3 h, and produced 28% dipicolinic acid more comparable to Bacillus licheniformis. Also they could produce 0.69 mg heat resistance protein after 1.5 h treatment in 100°C. The results showed that this strain could have biotechnological applications.Conflict of interests: The authors declare no conflict of interest.

  5. Coat protein-mediated resistance against an Indian isolate of the Cucumber mosaic virus subgroup IB in Nicotiana benthamiana

    Indian Academy of Sciences (India)

    A Srivastava; S K Raj

    2008-06-01

    Coat protein (CP)-mediated resistance against an Indian isolate of the Cucumber mosaic virus (CMV) subgroup IB was demonstrated in transgenic lines of Nicotiana benthamiana through Agrobacterium tumefaciens-mediated transformation. Out of the fourteen independently transformed lines developed, two lines were tested for resistance against CMV by challenge inoculations. The transgenic lines exhibiting complete resistance remained symptomless throughout life and showed reduced or no virus accumulation in their systemic leaves after virus challenge. These lines also showed virus resistance against two closely related strains of CMV. This is the first report of CP-mediated transgenic resistance against a CMV subgroup IB member isolated from India.

  6. Influence of coat protein transgene copy number on resistance in transgenic line 63-1 against Papaya ringspot virus isolates

    NARCIS (Netherlands)

    Souza, M.T.; Níckel, O.; Gonsalves, D.

    2005-01-01

    Line 63-1 is a 'Sunset'-derived transgenic papaya expressing the coat protein (CP) gene from a mild mutant of a Hawaiian isolate of Papaya ringspot virus (PRSV). Previous work showed that line 63-1 R, plants exhibited a range of resistance to severe PRSV isolates from Hawaii (HA), Jamaica (JA),

  7. Influence of coat protein transgene copy number on resistance in transgenic line 63-1 against Papaya ringspot virus isolates

    NARCIS (Netherlands)

    Souza, M.T.; Níckel, O.; Gonsalves, D.

    2005-01-01

    Line 63-1 is a 'Sunset'-derived transgenic papaya expressing the coat protein (CP) gene from a mild mutant of a Hawaiian isolate of Papaya ringspot virus (PRSV). Previous work showed that line 63-1 R, plants exhibited a range of resistance to severe PRSV isolates from Hawaii (HA), Jamaica (JA), Thai

  8. Ribosomal protein L3 mutations are associated with cfr-mediated linezolid resistance in clinical isolates of Staphylococcus cohnii.

    Science.gov (United States)

    Xu, Hongtao; Tian, Rui; Li, Yanming; Chen, Dongke; Liu, Yalin; Hu, Yunjian; Xiao, Fei

    2015-06-01

    From June, 2012 to November, 2013 five linezolid-resistant Staphylococcus cohnii isolates were identified in our hospital in Beijing, China. The investigation of the resistance mechanisms confirmed that the cfr-carrying plasmids were the main cause of linezolid resistance in those clinical isolates. Moreover, all the five isolates had ribosomal protein L3 mutations, which had different coordinate effect on cfr-mediated linezolid resistance directly through the substitution of serine 158 by phenylalanine or tyrosine in L3 protein. In this study, two types of plasmids (p432, p438) (Accession No. KM114207) were found, which share high sequence identity with previously reported cfr-carrying pRM01 and pMHZ plasmids originated from northern and southern China, showing wide regional dissemination in China. The stability of linezolid resistance was studied by passaging single colonies serially on antibiotic-free blood medium, which showed that the susceptible derivatives emerged until the passages 39-42 with the elimination of cfr-carrying plasmid. Thus the high stability of this plasmid may pose a risk for the transmission among patients or even cause an outbreak in clinical settings.

  9. In vitro resistance to human platelet microbicidal protein among urethral staphylococcal and enterococcal isolates with its correlation with prostatitis

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    Ivanov I

    2005-01-01

    Full Text Available The study was carried out to test the in vitro activity of human platelet microbicidal protein (hPMP on most commonly isolated urethral pathogens and compare the same with clinical isolates from cases of chronic prostatitis (CP. Urethral isolates of Staphylococcus aureus (n=19, coagulase negative staphylococci (n=40 and Enterococcus faecalis (n=16 from patients with or without CP were tested. The hPMP susceptibility of bacterial strains was determined by exposing bacterial cells to serial dilutions of hPMP. A significantly higher proportion of CP-strains of coagulase negative staphylococci (91.3% vs 5.88% was resistant to hPMP than was that of non-CP strains (P < 0.001. Among CP-strains of S.aureus studied, 77.8% were considered resistant to the bactericidal action of hPMP. All nine CP-strains of E.faecalis were highly resistant to hPMP. Most non-CP urethral isolates of S.aureus , coagulase negative staphylococci and E.faecalis were susceptible to the bactericidal action of hPMP, while CP isolates of all species were significantly more resistant to hPMP. Data from the present study may have significant implications in understanding the pathogenesis of CP.

  10. Outer Membrane Protein D Gene in Clinical Isolates of Pseudomonas Aeruginosa and its Role in Antibiotic Resistance

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    Neda Motaghi

    2016-03-01

    Full Text Available Background & Objectives: Pseudomonas aeruginosa is a common cause of nosocomial infection. OprD protein is a specific protein regulating the uptake of carbapenem antibiotic. Loss of OprD is the main mechanism of Pseudomonas Aeruginosa resistance to carbapenem. In this study, the presence of OprD gene is investigated in isolated Pseudomonas Aeruginosa in burn patients of Ghotboddin hospital in Shiraz. Material & Methods: 66 Pseudomonas Aeruginosa were isolated from wound specimens of 250 burn patients. Strain characteristics were confirmed by biochemical tests. Antibiogram was done via disc diffusion method. Finally, OprD gene was investigated by PCR. Results: Isolated Pseudomonas Aeruginosa showed more sensitivity to chloramphenicol and colicitin and more resistance  to ciprofloxacin, gentamycin, cefotaxim, ceftazidin, imipenem, meropenem, and erythromycin. 61 percent of isolates were positive for OprD gene by PCR. Conclusion: The findings of this study revealed that Colicitin and chloramphenicol are more effective in treatment of Pseudomonas Aeruginosa infections in burn patients, and deletion and mutation in OprD gene cause bacterium resistance to carbapenem antibiotic.

  11. A novel small acid soluble protein variant is important for spore resistance of most Clostridium perfringens food poisoning isolates.

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    Jihong Li

    2008-05-01

    Full Text Available Clostridium perfringens is a major cause of food poisoning (FP in developed countries. C. perfringens isolates usually induce the gastrointestinal symptoms of this FP by producing an enterotoxin that is encoded by a chromosomal (cpe gene. Those typical FP strains also produce spores that are extremely resistant to food preservation approaches such as heating and chemical preservatives. This resistance favors their survival and subsequent germination in improperly cooked, prepared, or stored foods. The current study identified a novel alpha/beta-type small acid soluble protein, now named Ssp4, and showed that sporulating cultures of FP isolates producing resistant spores consistently express a variant Ssp4 with an Asp substitution at residue 36. In contrast, Gly was detected at Ssp4 residue 36 in C. perfringens strains producing sensitive spores. Studies with isogenic mutants and complementing strains demonstrated the importance of the Asp 36 Ssp4 variant for the exceptional heat and sodium nitrite resistance of spores made by most FP strains carrying a chromosomal cpe gene. Electrophoretic mobility shift assays and DNA binding studies showed that Ssp4 variants with an Asp at residue 36 bind more efficiently and tightly to DNA than do Ssp4 variants with Gly at residue 36. Besides suggesting one possible mechanistic explanation for the highly resistant spore phenotype of most FP strains carrying a chromosomal cpe gene, these findings may facilitate eventual development of targeted strategies to increase killing of the resistant spores in foods. They also provide the first indication that SASP variants can be important contributors to intra-species (and perhaps inter-species variations in bacterial spore resistance phenotypes. Finally, Ssp4 may contribute to spore resistance properties throughout the genus Clostridium since ssp4 genes also exist in the genomes of other clostridial species.

  12. A novel small acid soluble protein variant is important for spore resistance of most Clostridium perfringens food poisoning isolates.

    Science.gov (United States)

    Li, Jihong; McClane, Bruce A

    2008-05-02

    Clostridium perfringens is a major cause of food poisoning (FP) in developed countries. C. perfringens isolates usually induce the gastrointestinal symptoms of this FP by producing an enterotoxin that is encoded by a chromosomal (cpe) gene. Those typical FP strains also produce spores that are extremely resistant to food preservation approaches such as heating and chemical preservatives. This resistance favors their survival and subsequent germination in improperly cooked, prepared, or stored foods. The current study identified a novel alpha/beta-type small acid soluble protein, now named Ssp4, and showed that sporulating cultures of FP isolates producing resistant spores consistently express a variant Ssp4 with an Asp substitution at residue 36. In contrast, Gly was detected at Ssp4 residue 36 in C. perfringens strains producing sensitive spores. Studies with isogenic mutants and complementing strains demonstrated the importance of the Asp 36 Ssp4 variant for the exceptional heat and sodium nitrite resistance of spores made by most FP strains carrying a chromosomal cpe gene. Electrophoretic mobility shift assays and DNA binding studies showed that Ssp4 variants with an Asp at residue 36 bind more efficiently and tightly to DNA than do Ssp4 variants with Gly at residue 36. Besides suggesting one possible mechanistic explanation for the highly resistant spore phenotype of most FP strains carrying a chromosomal cpe gene, these findings may facilitate eventual development of targeted strategies to increase killing of the resistant spores in foods. They also provide the first indication that SASP variants can be important contributors to intra-species (and perhaps inter-species) variations in bacterial spore resistance phenotypes. Finally, Ssp4 may contribute to spore resistance properties throughout the genus Clostridium since ssp4 genes also exist in the genomes of other clostridial species.

  13. Isolation and Cloning of cDNA Fragment of Gene Encoding for Multidrug Resistance Associated Protein from M. affine.

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    Utut Widyastuti Suharsono

    2008-11-01

    Full Text Available Isolation and Cloning of cDNA Fragment of Gene Encoding for Multidrug Resistance Associated Protein from M. affine. M. affine can grow well in acid soil with high level of soluble aluminum. One of the important proteins in the detoxifying xenobiotic stress including acid and Al stresses is a multidrug resistance associated protein (MRP encoded by mrp gene. The objective of this research is to isolate and clone the cDNA fragment of MaMrp encoding MRP from M. affine. By reverse transcription, total cDNA had been synthesized from the total RNA as template. The fragment of cDNA MaMrp had been successfully isolated by PCR by using total cDNA as template and mrp primer designed from A. thaliana, yeast, and human. This fragment was successfully inserted into pGEM-T Easy and the recombinant plasmid was successfully introduced into E. coli DH5α. Nucleotide sequence analysis showed that the lenght of MaMrp fragment is 633 bp encoding 208 amino acids. Local alignment analysis based on nucleotide of mRNA showed that MaMrp fragment is 69% identical to AtMrp1 and 63% to AtMrp from A. thaliana. Based on deduced amino acid sequence, MaMRP is 84% identical to part of AtMRP13, 77% to AtMRP12, and 73% to AtMRP1 from A. thaliana respectively. Alignment analysis with AtMRP1 showed that MaMRP fragment is located in TM1 and NBF1 domains and has a specific amino acid sequence QCKAQLQNMEEE.

  14. Protein Profiling on Meglumine Antimoniate (Glucantime® Sensitive and Resistant L. tropica Isolates by 2- Dimentional Gel Electrophoresis: A Preliminary Study

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    R Hadighi

    2009-02-01

    Full Text Available Background: Glucantime® is the first- line drug for the treatment of all forms of leishmaniasis. Unfortunately, the prevalence of parasites becoming resistant to Glucantime® is increasing in several parts of the world including Iran. As protein is the most important target for drugs in response to a variety of signals including drugs so, it seems expression protein patterns in sensitive and resistant Leishmania parasites could greatly help us about the mechanisms of responses to antileishmanial drugs. In this study, we used 2-dimentional gel electrophoresis (2-DE method to determine protein expression profiles between drug (Glucantime® sensitive and resistant Leishmania tropica isolated from Iranian an­throponotic cutaneous leishmaniasis (ACL patients."nMethods: We used from the two confirmed genetically of Glucantime® sensitive (Mash-4 and resistant (Mash-927 field strains of L. tropica, isolated from ACL patients in north eastern Iran. The two Leishmania isolates were cultured, promastigotes were harvested followed by protein extraction using TCA/Aceton to study protein profiling, 2-DE was done and gels stained with silver nitrate."nResults: At least 2236 distinct protein spots were detected. Twelve spots out of them, showed significant changes in expression in resistant compared to sensitive isolates. Of these, 11 protein spots were up- and one was down-regulated."nConclusions: This preliminary study has showed that a number of proteins differentially expressed in drug (Glucan­time® resistance L. tropica and probably the role of these proteins are increasing the parasite resistance against the drug and delay in cell death.

  15. A new enabling proteomics methodology to investigate membrane associated proteins from parasitic nematodes: case study using ivermectin resistant and ivermectin susceptible isolates of Caenorhabditis elegans and Haemonchus contortus.

    Science.gov (United States)

    Hart, Elizabeth H; Brophy, Peter M; Prescott, Mark; Bartley, David J; Wolf, Basil T; Hamilton, Joanne V

    2015-01-30

    The mechanisms involved in anthelmintic resistance (AR) are complex but a greater understanding of AR management is essential for effective and sustainable control of parasitic helminth worms in livestock. Current tests to measure AR are time consuming and can be technically problematic, gold standard diagnostics are therefore urgently required to assist in combatting the threat from drug resistant parasites. For anthelmintics such as ivermectin (IVM), target proteins may be present in the cellular membrane. As proteins usually act in complexes and not in isolation, AR may develop and be measurable in the target associated proteins present in the parasite membrane. The model nematode Caenorhabditis elegans was used to develop a sub-proteomic assay to measure protein expression differences, between IVM resistant and IVM susceptible isolates in the presence and absence of drug challenge. Evaluation of detergents including CHAPS, ASB-14, C7BzO, Triton ×100 and TBP (tributyl phosphine) determined optimal conditions for the resolution of membrane proteins in Two Dimensional Gel Electrophoresis (2DE). These sub-proteomic methodologies were then translated and evaluated using IVM-susceptible and IVM-resistant Haemonchus contortus; a pathogenic blood feeding parasitic nematode which is of global importance in livestock health, welfare and productivity. We have demonstrated the successful resolution of membrane associated proteins from both C. elegans and H. contortus isolates, using a combination of CHAPS and the zwitterionic amphiphilic surfactant ASB-14 to further support the detection of markers for AR. Copyright © 2014. Published by Elsevier B.V.

  16. Effect of soy isolate protein and resistance exercises on muscle performance and bone health of osteopenic/osteoporotic post-menopausal women.

    Science.gov (United States)

    Shenoy, Shweta; Bedi, Reecha; Sandhu, Jaspal S

    2013-01-01

    There are contradictory reports regarding the effect of soy protein isolate on bone health in menopause. The main objective of this study was to assess the influence of soy isolate protein intake and resistance exercises on isokinetic muscle strength, endurance, power, and bone health parameters in osteopenic/osteoporotic postmenopausal women. Sixty osteoporotic sedentary women (mean age 54.55 years) were randomly assigned to three groups: soy isolate protein (Group A), soy + exercise group (Group B), and control group (Group C). Group B performed supervised progressive resistance exercises 4 times/week for 12 weeks. Muscle performance was measured by isokinetic dynamometry, and bone health was measured by ultrasound densitometry. Analysis of variance showed significant bone and muscle strength gains (p < .05) both in Group A and B, with the improvements more pronounced in Group B. Significant muscle performance changes, after intervention, were evident and bone strength increases may parallel changes in muscle strength.

  17. Isolation of Escherichia coli rpoB mutants resistant to killing by lambda cII protein and altered in pyrE gene attenuation

    DEFF Research Database (Denmark)

    Hammer, Karin; Jensen, Kaj Frank; Poulsen, Peter;

    1987-01-01

    Escherichia coli mutants simultaneously resistant to rifampin and to the lethal effects of bacteriophage lambda cII protein were isolated. The sck mutant strains carry alterations in rpoB that allow them to survive cII killing (thus the name sck), but that do not impair either the expression of c...

  18. Detection of amino acid substitutions in the GyrA protein of fluoroquinolone-resistant typhoidal Salmonella isolates using high-resolution mass spectrometry.

    Science.gov (United States)

    Hassing, Robert-Jan; Goessens, Wil H; Zeneyedpour, Lona; Sultan, Sadaf; van Kampen, Jeroen J; Verbon, Annelies; van Genderen, Perry J; Hays, John P; Luider, Theo M; Dekker, Lennard J

    2016-05-01

    Infections with typhoidal Salmonella isolates that are resistant to fluoroquinolone antibiotics have become very common in several Asian countries. In the majority of these cases, resistance to fluoroquinolone-based antibiotics is associated with genetic mutations in the quinolone resistance-determining region (QRDR) of the bacterial DNA gyrase gene gyrA. The objective of this study was to detect these amino acid substitutions by high-resolution mass spectrometry instead of sequencing of the gyrA gene. A liquid chromatography-mass spectrometry (LC-MS) methodology was developed and evaluated for the detection of amino acid substitutions in the GyrA protein of 23 typhoidal Salmonella isolates. These isolates included typhoidal Salmonella that possessed different antibiotic sensitivities to fluoroquinolone antibiotics. The LC-MS methodology correctly identified peptide sequences associated with phenotypic QRDR mutations of the GyrA protein in all 23 phenotypically diverse typhoidal Salmonella isolates tested. In conclusion, a reliable and rapid LC-MS methodology has been developed that is able to identify GyrA QRDR mutations that are involved in the development of fluoroquinolone resistance in typhoidal Salmonella spp. Furthermore, this 'proof of principle' study indicates the potential usefulness of LC-MS in future detection of antibiotic resistance. Copyright © 2016 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  19. Protein loss during nuclear isolation

    OpenAIRE

    1983-01-01

    Cryomicrodissection makes possible the measurement of the entire in vivo protein content of the amphibian oocyte nucleus and provides a heretofore missing baseline for estimating protein loss during nuclear isolation by other methods. When oocyte nuclei are isolated into an aqueous medium, they lose 95% of their protein with a half-time of 250 s. This result implies an even more rapid loss of protein from aqueously isolated nuclei of ordinary-size cells.

  20. One of two TolC-like proteins is involved in antibiotic resistance and biofilm formation of Actinobacillus pleuropneumoniae clinical isolate SC1516

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    Ying Li

    2016-10-01

    Full Text Available Actinobacillus pleuropneumoniae is the etiologic agent of porcine contagious pleuropneumonia, a significant disease that causes serious economic losses to the swine industry worldwide. Persistent infections caused by bacterial biofilms are recalcitrant to treat because of the particular drug resistance of biofilm-dwelling cells. TolC, a key component of multidrug efflux pumps, are responsible for multidrug resistance in many Gram-negative bacteria. In this study, we identified two TolC-like proteins, TolC1 and TolC2, in A. pleuropneumoniae. Deletion of tolC1, but not tolC2, caused a significant reduction in biofilm formation, as well as increased drug sensitivity of both planktonic and biofilm cells. The genetic-complementation of the tolC1 mutation restored the competent biofilm and drug resistance. Besides, biofilm formation was inhibited and drug sensitivity was increased by the addition of phenylalanine-arginine beta-naphthylamide (PAβN, a well-known efflux pump inhibitor (EPI, suggesting a role for EPI in antibacterial strategies towards drug tolerance of A. pleuropneumoniae. Taken together, TolC1 is required for biofilm formation and is a part of the multidrug resistance machinery of both planktonic and biofilm cells, which could supplement therapeutic strategies for resistant bacteria and biofilm-related infections of A. pleuropneumoniae clinical isolate SC1516.

  1. Multidrug resistance associated proteins in multidrug resistance

    OpenAIRE

    Sodani, Kamlesh; Patel, Atish; Kathawala, Rishil J.; Chen, Zhe-Sheng

    2012-01-01

    Multidrug resistance proteins (MRPs) are members of the C family of a group of proteins named ATP-binding cassette (ABC) transporters. These ABC transporters together form the largest branch of proteins within the human body. The MRP family comprises of 13 members, of which MRP1 to MRP9 are the major transporters indicated to cause multidrug resistance in tumor cells by extruding anticancer drugs out of the cell. They are mainly lipophilic anionic transporters and are reported to transport fr...

  2. Generation of a Vancomycin-Resistant Enterococcus faecium Clinical Isolate Expressing a (FMN)-Based Fluorescent Protein

    OpenAIRE

    PINILLA REDONDO, RAFAEL

    2014-01-01

    [EN] Infections caused by multidrug-resistant (MDR) bacteria, including Vancomycin-Resistant Enterococcus (VRE), have become one of the greatest clinical challenges of the 21st century. Particularly, VRE strains of E. faecium, a natural member of the gastrointestinal consortia, have recently emerged as one of the most problematic cases of MDR nosocomial pathogens. It is widely known that the establishment of high-level intestinal colonization of this bacterium, triggered by antibiotic treatme...

  3. Binding site alteration is responsible for field-isolated resistance to Bacillus thuringiensis Cry2A insecticidal proteins in two Helicoverpa species.

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    Silvia Caccia

    Full Text Available BACKGROUND: Evolution of resistance by target pests is the main threat to the long-term efficacy of crops expressing Bacillus thuringiensis (Bt insecticidal proteins. Cry2 proteins play a pivotal role in current Bt spray formulations and transgenic crops and they complement Cry1A proteins because of their different mode of action. Their presence is critical in the control of those lepidopteran species, such as Helicoverpa spp., which are not highly susceptible to Cry1A proteins. In Australia, a transgenic variety of cotton expressing Cry1Ac and Cry2Ab (Bollgard II comprises at least 80% of the total cotton area. Prior to the widespread adoption of Bollgard II, the frequency of alleles conferring resistance to Cry2Ab in field populations of Helicoverpa armigera and Helicoverpa punctigera was significantly higher than anticipated. Colonies established from survivors of F(2 screens against Cry2Ab are highly resistant to this toxin, but susceptible to Cry1Ac. METHODOLOGY/PRINCIPAL FINDINGS: Bioassays performed with surface-treated artificial diet on neonates of H. armigera and H. punctigera showed that Cry2Ab resistant insects were cross-resistant to Cry2Ae while susceptible to Cry1Ab. Binding analyses with (125I-labeled Cry2Ab were performed with brush border membrane vesicles from midguts of Cry2Ab susceptible and resistant insects. The results of the binding analyses correlated with bioassay data and demonstrated that resistant insects exhibited greatly reduced binding of Cry2Ab toxin to midgut receptors, whereas no change in (125I-labeled-Cry1Ac binding was detected. As previously demonstrated for H. armigera, Cry2Ab binding sites in H. punctigera were shown to be shared by Cry2Ae, which explains why an alteration of the shared binding site would lead to cross-resistance between the two Cry2A toxins. CONCLUSION/SIGNIFICANCE: This is the first time that a mechanism of resistance to the Cry2 class of insecticidal proteins has been reported

  4. methicillin resistance in staphylococcal isolates from

    African Journals Online (AJOL)

    to the cheap, readily available brand-spectrum antibiotics; ampicllin, amoxicillin, chloramphenicol, tetracycline and ... Analysis of the multiple antibiotic resistance (MAR) index of isolates and the production of - ... microorganisms, thus infection.

  5. Tiamulin resistance in porcine Brachyspira pilosicoli isolates.

    Science.gov (United States)

    Pringle, M; Landén, A; Franklin, A

    2006-02-01

    There are few studies on antimicrobial susceptibility of Brachyspira pilosicoli, therefore this study was performed to investigate the situation among isolates from pigs. The tiamulin and tylosin susceptibility was determined by broth dilution for 93 and 86 porcine B. pilosicoli isolates, respectively. The isolates came from clinical samples taken in Swedish pig herds during the years 2002 and 2003. The tylosin minimal inhibitory concentration (MIC) was >16 microg/ml for 50% (n=43) of the isolates tested. A tiamulin MIC >2 microg/ml was obtained for 14% (n=13) of the isolates and these were also tested against doxycycline, salinomycin, valnemulin, lincomycin and aivlosin. For these isolates the susceptibility to salinomycin and doxycycline was high but the MICs for aivlosin varied. The relationship between the 13 tiamulin resistant isolates was analyzed by pulsed-field gel electrophoresis (PFGE). Among the 13 isolates 10 different PFGE patterns were identified.

  6. Yeast ABC proteins involved in multidrug resistance.

    Science.gov (United States)

    Piecuch, Agata; Obłąk, Ewa

    2014-03-01

    Pleiotropic drug resistance is a complex phenomenon that involves many proteins that together create a network. One of the common mechanisms of multidrug resistance in eukaryotic cells is the active efflux of a broad range of xenobiotics through ATP-binding cassette (ABC) transporters. Saccharomyces cerevisiae is often used as a model to study such activity because of the functional and structural similarities of its ABC transporters to mammalian ones. Numerous ABC transporters are found in humans and some are associated with the resistance of tumors to chemotherapeutics. Efflux pump modulators that change the activity of ABC proteins are the most promising candidate drugs to overcome such resistance. These modulators can be chemically synthesized or isolated from natural sources (e.g., plant alkaloids) and might also be used in the treatment of fungal infections. There are several generations of synthetic modulators that differ in specificity, toxicity and effectiveness, and are often used for other clinical effects.

  7. Island cotton Gbve1 gene encoding a receptor-like protein confers resistance to both defoliating and non-defoliating isolates of Verticillium dahliae.

    Directory of Open Access Journals (Sweden)

    Baolong Zhang

    Full Text Available Verticillium wilt caused by soilborne fungus Verticillium dahliae could significantly reduce cotton yield. Here, we cloned a tomato Ve homologous gene, Gbve1, from an island cotton cultivar that is resistant to Verticillium wilt. We found that the Gbve1 gene was induced by V. dahliae and by phytohormones salicylic acid, jasmonic acid, and ethylene, but not by abscisic acid. The induction of Gbve1 in resistant cotton was quicker and stronger than in Verticillium-susceptible upland cotton following V. dahliae inoculation. Gbve1 promoter-driving GUS activity was found exclusively in the vascular bundles of roots and stems of transgenic Arabidopsis. Virus-induced silencing of endogenous genes in resistant cotton via targeting a fragment of the Gbve1 gene compromised cotton resistance to V. dahliae. Furthermore, we transformed the Gbve1 gene into Arabidopsis and upland cotton through Agrobacterium-mediated transformation. Overexpression of the Gbve1 gene endowed transgenic Arabidopsis and upland cotton with resistance to high aggressive defoliating and non-defoliating isolates of V. dahliae. And HR-mimic cell death was observed in the transgenic Arabidopsis. Our results demonstrate that the Gbve1 gene is responsible for resistance to V. dahliae in island cotton and can be used for breeding cotton varieties that are resistant to Verticillium wilt.

  8. Multidrug resistance associated proteins in multidrug resistance

    Institute of Scientific and Technical Information of China (English)

    Kamlesh Sodani; Atish Patel; Rishil J. Kathawala; Zhe-Sheng Chen

    2012-01-01

    Multidrug resistance proteins (MRPs) are members of the C family of a group of proteins named ATP-binding cassette (ABC) transporters.These ABC transporters together form the largest branch of proteins within the human body.The MRP family comprises of 13 members,of which MRP1 to MRP9 are the major transporters indicated to cause multidrug resistance in tumor cells by extruding anticancer drugs out of the cell.They are mainly lipophilic anionic transporters and are reported to transport free or conjugates of glutathione (GSH),glucuronate,or sulphate.In addition,MRP1 to MRP3 can transport neutral organic drugs in free form in the presence of free GSH.Collectively,MRPs can transport drugs that differ structurally and mechanistically,including natural anticancer drugs,nucleoside analogs,antimetabolites,and tyrosine kinase inhibitors.Many of these MRPs transport physiologically important anions such as leukotriene C4,bilirubin glucuronide,and cyclic nucleotides.This review focuses mainly on the physiological functions,cellular resistance characteristics,and probable in vivo role of MRP1 to MRP9.

  9. [Phenotypic and genotypic characterization of imipenem-resistant Pseudomonas aeruginosa isolated in a Buenos Aires hospital].

    Science.gov (United States)

    Cejas, D; Almuzara, M; Santella, G; Tuduri, A; Palombarani, S; Figueroa, S; Gutkind, G; Radice, M

    2008-01-01

    From 129 P. aeruginosa isolated at a health care centre located in Buenos Aires (Hospital "Eva Perón"), 14% produced IMP-13. Although 18 isolates were metallo-beta-lactamases (MBL) producers, only those isolates that displayed altered outer membrane protein profiles correlated with the resistant category according to CLSI or even Subcomisión de Antimicrobianos, SADEBAC, AAM. Phenotypic screening of metallo-beta-lactamases proved to be appropriate for detecting MBL producing isolates. IMP-13 producing isolates corresponded to at least five different clonal types, which not only suggests the dissemination of the resistant strain but also of the resistant marker.

  10. In vitro antibacterial activity of venom protein isolated from sea snake Enhydrina schistosa against drug-resistant human pathogenic bacterial strains

    Institute of Scientific and Technical Information of China (English)

    Palani Damotharan; Anguchamy Veeruraj; Muthuvel Arumugam; Thangavel Balasubramanian

    2015-01-01

    Objective:To evaluate the antibacterial activity of sea snake (Enhydrina schistosa) venom protein against drug-resistant human pathogenic bacterial strains. Methods:The venom was collected by milking process from the live specimens of sea snake are using capillary tubes or glass plates. Venom was purified by ion exchange chromatography and it was tested for in-vitro antibacterial activity against 10 drug-resistant human pathogenic bacterial strains using the standard disc diffusion method. Results:The notable antibacterial activity was observed at 150 µg/mL concentration of purified venom and gave its minimum inhibitory concentrations values exhibited between 200-100 µg/mL against all the tested bacterial strains. The maximum zone of inhibition was observed at 16.4 mm against Salmonella boydii and the minimum activity was observed at 7.5 mm against Pseudomonas aeruginosa. After the sodium-dodecyl-sulfate-polyacrylamide gel electrophoresis there were a clear single band was detected in the gel that corresponding to purified venom protein molecular weight of 44 kDa. Conclusions:These results suggested that the sea snake venom might be a feasible source for searching potential antibiotics agents against human pathogenic diseases.

  11. Understanding the differences in gelling properties between lupin protein isolate and soy protein isolate

    NARCIS (Netherlands)

    Berghout, J.A.M.; Boom, R.M.; Goot, van der A.J.

    2015-01-01

    The gelling properties of lupin protein isolate (LPI) were compared with those of soy protein isolate (SPI). It was found that LPI behaves fundamentally different than SPI, evidenced by the formation of weaker and deformable gels. Further investigation shows that both protein isolates can be conside

  12. Diversity in fosfomycin resistance proteins

    Directory of Open Access Journals (Sweden)

    Matthew K. Thompson

    2015-03-01

    Full Text Available Certain strains of the soil microorganism Streptomyces produce an antibiotic, fosfomycin [(1 R,2 S-epoxypropylphosphonic acid], which is effective against both Gram-positive and Gram-negative pathogens by inhibiting the first committed step in cell-wall biosynthesis. Fosfomycin resistance proteins are metallo-enzymes that are known to inactivate the antibiotic by the addition of nucleophiles such as water, glutathione (GSH, l-cysteine and bacillithiol (BSH to the oxirane ring of the molecule. Progress in the characterisation of FosB-type fosfomycin resistance proteins found in many Gram-positive organisms has been slow. This paper provides a brief description of the diversity of fosfomycin resistance proteins in general and, more specifically, new data characterising the substrate selectivity, structure, mechanism and metal-ion dependence of FosB enzymes from pathogenic strains of Staphylococcus and Bacillus. These new findings include the high-resolution X-ray diffraction structures of FosB enzymes from Staphylococcus aureus and Bacillus cereus in various liganded states and kinetic data that suggest that Mn(II and BSH are the preferred divalent cation and thiol substrate for the reaction, respectively. The discovery of the inhibition of the enzyme by Zn(II led to the determination of a ternary structure of the FosB·Zn(II·fosfomycin·l-Cys complex which reveals both substrates present in a pose prior to reaction.

  13. Isolation of plant cell wall proteins

    OpenAIRE

    Jamet, Elisabeth; Boudart, Georges; Borderies, Gisèle; Charmont, Stéphane; Lafitte, Claude; Rossignol, Michel; Canut, Hervé; Pont-Lezica, Rafael F

    2007-01-01

    The quality of a proteomic analysis of a cell compartment strongly depends on the reliability of the isolation procedure for the cell compartment of interest. Plant cell walls possess specific drawbacks: (i) the lack of a surrounding membrane may result in the loss of cell wall proteins (CWP) during the isolation procedure; (ii) polysaccharide networks of cellulose, hemicelluloses and pectins form potential traps for contaminants such as intracellular proteins; (iii) the presence of proteins ...

  14. Protein distribution in lupin protein isolates from Lupinus angustifolius L. prepared by various isolation techniques.

    Science.gov (United States)

    Muranyi, Isabel S; Volke, Daniela; Hoffmann, Ralf; Eisner, Peter; Herfellner, Thomas; Brunnbauer, Markus; Koehler, Peter; Schweiggert-Weisz, Ute

    2016-09-15

    Differences in the protein distribution of various protein isolates from Lupinus angustifolius L. Vitabor were identified as affected by the isolation procedure (alkaline and/or salt-induced extraction followed by isoelectric and/or dilutive precipitation). Protein isolates extracted in alkaline solution showed higher protein yields (26.4-31.7%) compared to salt-induced extraction (19.8-30.0%) or combined alkaline and salt-induced extraction (23.3-25.6%). Chemical variations among the protein isolates especially occurred within the albumins. Protein isolates precipitated isoelectrically showed the highest contents, whereas protein isolates precipitated by dilutive showed the lowest contents of conglutin δ. Furthermore, the alkaline subunits of conglutin α and conglutin γ decreased during alkaline extraction compared to salt-induced extraction. A decrease in protein-bound polar and basic amino acids was shown after protein isolation. In contrast, the amounts of nonpolar, aliphatic, aromatic, hydroxylated and sulfur-rich amino acids were higher in the lupin protein isolates compared to the lupin flakes. However, the functional side chains could not be related to the specific molecular arrangements of the protein isolates, as a similar amino acid composition was found among the protein isolates. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Stiffening in gels containing whey protein isolate

    NARCIS (Netherlands)

    Purwanti, N.; Veen, van der E.; Goot, van der A.J.; Boom, R.M.

    2013-01-01

    Gels made only from whey protein isolate (WPI) stiffened over the first few days of storage, after which the textural properties remained nearly constant. However, protein gels containing WPI microparticles, at the same total protein content, stiffened over a longer period than those without micropa

  16. RHEOLOGY OF EXTRUDED WHEY PROTEIN ISOLATE

    Science.gov (United States)

    Whey protein isolate (WPI), a high-quality protein used to fortify a number of foods, may be texturized with a twin-screw extruder. Since extrusion of food is commonly performed above 70°C, which causes whey protein to denature, cold extrusion below 70°C was investigated to determine the effects on...

  17. Isolation and Characterization of Nickel Uptake by Nickel Resistant Bacterial Isolate (NiRBI)

    Institute of Scientific and Technical Information of China (English)

    JAGDISH S PATEL; PRERNA C PATEL; KIRAN KALIA

    2006-01-01

    Bioremediation technology has gained importance because microbes could be the convenient source of bio-absorption/bioaccumulation of metals from effluent streams. Methods The nickel-resistant bacterial isolates (NiRBI)were selected from various bacterial isolates from industrial effluent and grown in nutrient broth containing different concentrations of nickel sulfate (0.3-3.0 mmol/L) and their capability of accumulating metal from the medium. Results Well-defined growth of NiRBI was observed in the medium containing up to 2.5 mmol/L of nickel. The isolate was identified using 16S rRNA and closely related to Pseudomonas fragi. Maximum accumulation of nickel (0.59 mg/g dry weight of bacterial cells) was observed when NiRBI was grown in media containing 2 mmol/L of nickel. The protein profile of the NiRBI cellular extract by SDS-PAGE showed two metal stress-induced proteins of molecular weight 48 KD and 18 KD with a simultaneous down regulation of four proteins of 46.7 KD, 42.2 KD, 19.7 KD, and 4.0 KD. Conclusion 48 KD and 18 KD proteins play a role in metal resistance mechanism by NiRBI.

  18. Linezolid resistance in Enterococcus faecium isolated in Ontario, Canada.

    Science.gov (United States)

    Patel, Samir N; Memari, Nader; Shahinas, Dea; Toye, Baldwin; Jamieson, Frances B; Farrell, David J

    2013-12-01

    Recent studies have described linezolid-resistant MRSA and vancomycin-resistant enterococci (VRE) occurring worldwide, including an outbreak of linezolid-resistant MRSA. The objective of this study was to determine if linezolid-resistant enterococci are present in clinical isolates in Ontario, Canada. From January 2010 to June 2012, all enterococcal isolates submitted to the Public Health Ontario Laboratory (PHOL) for confirmation of VRE and susceptibility testing were included in this study. Of 2829 enterococcal isolates tested, 12 Enterococcus faecium were found to be resistant to linezolid. All linezolid-resistant isolates were also resistant to ampicillin, ciprofloxacin, and vancomycin. In addition, 33% of isolates were non-susceptible to daptomycin, whereas 41% were resistant to quinupristin/dalfopristin. Molecular characterization of these isolates showed that 8/12 isolates (66.7%) contained the mutation G2576T in 23S rRNA, which has been associated with linezolid resistance. Amplification and sequencing of L3- and L4-coding genes did not reveal mutations associated with linezolid resistance. One isolate contained the cfr gene, which is associated with linezolid resistance, and has been found in staphylococcal species and E. faecalis. These data show that occurrence of linezolid resistance is still rare among enterococcal isolates referred to PHOL though detection of cfr in E. faecium is concerning as it has the potential to disseminate among other enterococci.

  19. Resistance to moist conditions of whey protein isolate and pea starch biodegradable films and low density polyethylene nondegradable films: a comparative study

    Science.gov (United States)

    Mehyar, G. F.; Bawab, A. Al

    2015-10-01

    Biodegradable packaging materials are degraded under the natural environmental conditions. Therefore using them could alleviate the problem of plastics accumulation in nature. For effective replacement of plastics, with biodegradable materials, biodegradable packages should keep their properties under the high relative humidity (RH) conditions. Therefore the objectives of the study were to develop biodegradable packaging material based on whey protein isolate (WPI) and pea starch (PS). To study their mechanical, oxygen barrier and solubility properties under different RHs compared with those of low density polyethylene (LDPE), the most used plastic in packaging. Films of WPI and PS were prepared separately and conditioned at different RH (30-90%) then their properties were studied. At low RHs ( 40% RH. Oxygen permeability of WPI and LDPE did not adversely affected by increasing RH to 65%. Furthermore, WPI and LDPE films had lower degree of hydration at 50% and 90% RH and total soluble matter than PS films. These results suggest that WPI could be successfully replacing LDPE in packaging of moist products.

  20. Mutational and acquired carbapenem resistance mechanisms in multidrug resistant Pseudomonas aeruginosa clinical isolates from Recife, Brazil

    Science.gov (United States)

    Cavalcanti, Felipe Lira de Sá; Mirones, Cristina Rodríguez; Paucar, Elena Román; Montes, Laura Álvarez; Leal-Balbino, Tereza Cristina; de Morais, Marcia Maria Camargo; Martínez-Martínez, Luis; Ocampo-Sosa, Alain Antonio

    2015-01-01

    An investigation was carried out into the genetic mechanisms responsible for multidrug resistance in nine carbapenem-resistant Pseudomonas aeruginosaisolates from different hospitals in Recife, Brazil. Susceptibility to antimicrobial agents was determined by broth microdilution. Polymerase chain reaction (PCR) was employed to detect the presence of genes encoding β-lactamases, aminoglycoside-modifying enzymes (AMEs), 16S rRNA methylases, integron-related genes and OprD. Expression of genes coding for efflux pumps and AmpC cephalosporinase were assessed by quantitative PCR. The outer membrane proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The blaSPM-1, blaKPC-2 and blaGES-1 genes were detected in P. aeruginosaisolates in addition to different AME genes. The loss of OprD in nine isolates was mainly due to frameshift mutations, premature stop codons and point mutations. An association of loss of OprD with the overexpression of MexAB-OprM and MexXY-OprM was observed in most isolates. Hyper-production of AmpC was also observed in three isolates. Clonal relationship of the isolates was determined by repetitive element palindromic-PCR and multilocus sequence typing. Our results show that the loss of OprD along with overexpression of efflux pumps and β-lactamase production were responsible for the multidrug resistance in the isolates analysed. PMID:26676375

  1. Isolation and characterization of arsenic resistant bacteria from wastewater

    Directory of Open Access Journals (Sweden)

    Syed Zaghum Abbas

    2014-12-01

    Full Text Available The present study proposed the isolation of arsenic resistant bacteria from wastewater. Only three bacterial isolates (MNZ1, MNZ4 and MNZ6 were able to grow in high concentrations of arsenic. The minimum inhibitory concentrations of arsenic against MNZ1, MNZ4 and MNZ6 were 300 mg/L, 300 mg/L and 370 mg/L respectively. The isolated strains showed maximum growth at 37 ºC and at 7.0 pH in control but in arsenite stress Luria Bertani broth the bacterial growth is lower than control. All strains were arsenite oxidizing. All strains were biochemically characterized and ribotyping (16S rRNA was done for the purpose of identification which confirmed that MNZ1 was homologous to Enterobacter sp. while MNZ4 and MNZ6 showed their maximum homology with Klebsiella pneumoniae. The protein profiling of these strains showed in arsenic stressed and non stressed conditions, so no bands of induced proteins appeared in stressed conditions. The bacterial isolates can be exploited for bioremediation of arsenic containing wastes, since they seem to have the potential to oxidize the arsenite (more toxic into arsenate (less toxic form.

  2. Isolation and characterization of arsenic resistant bacteria from wastewater.

    Science.gov (United States)

    Abbas, Syed Zaghum; Riaz, Mehwish; Ramzan, Naseem; Zahid, M Tariq; Shakoori, Farah R; Rafatullah, Mohd

    2014-01-01

    The present study proposed the isolation of arsenic resistant bacteria from wastewater. Only three bacterial isolates (MNZ1, MNZ4 and MNZ6) were able to grow in high concentrations of arsenic. The minimum inhibitory concentrations of arsenic against MNZ1, MNZ4 and MNZ6 were 300 mg/L, 300 mg/L and 370 mg/L respectively. The isolated strains showed maximum growth at 37 °C and at 7.0 pH in control but in arsenite stress Luria Bertani broth the bacterial growth is lower than control. All strains were arsenite oxidizing. All strains were biochemically characterized and ribotyping (16S rRNA) was done for the purpose of identification which confirmed that MNZ1 was homologous to Enterobacter sp. while MNZ4 and MNZ6 showed their maximum homology with Klebsiella pneumoniae. The protein profiling of these strains showed in arsenic stressed and non stressed conditions, so no bands of induced proteins appeared in stressed conditions. The bacterial isolates can be exploited for bioremediation of arsenic containing wastes, since they seem to have the potential to oxidize the arsenite (more toxic) into arsenate (less toxic) form.

  3. Detection of ST772 Panton-Valentine leukocidin-positive methicillin-resistant Staphylococcus aureus (Bengal Bay clone and ST22 S. aureus isolates with a genetic variant of elastin binding protein in Nepal

    Directory of Open Access Journals (Sweden)

    R.H. Pokhrel

    2016-05-01

    Full Text Available Genetic characteristics were analysed for recent clinical isolates of methicillin-resistant and -susceptible Staphylococcus aureus (MRSA and MSSA respectively in Kathmandu, Nepal. MRSA isolates harbouring Panton-Valentine leukocidin (PVL genes were classified into ST1, ST22 and ST88 with SCCmec-IV and ST772 with SCCmec-V (Bengal Bay clone, while PVL-positive MSSA into ST22, ST30 and ST772. ST22 isolates (PVL-positive MRSA and MSSA, PVL-negative MRSA possessed a variant of elastin binding protein gene (ebpS with an internal deletion of 180 bp, which was similar to that reported for ST121 S. aureus previously outside Nepal. Phylogenetic analysis indicated that the ebpS variant in ST22 might have occurred independently of ST121 strains. This is the first report of ST772 PVL-positive MRSA in Nepal and detection of the deletion variant of ebpS in ST22 S. aureus.

  4. Isolation rate and drug resistance patterns of Shigella species ...

    African Journals Online (AJOL)

    Isolation rate and drug resistance patterns of Shigella species among ... use of water from unprotected sources and absence of latrine were the risk factors that ... to identify changes in the prevalence and antimicrobial resistance patterns of ...

  5. Norfloxacin resistance in a clinical isolate of Escherichia coli.

    OpenAIRE

    Aoyama, H; Sato, K; Kato, T.; Hirai, K; Mitsuhashi, S.

    1987-01-01

    Analysis of DNA gyrase supercoiling and of norfloxacin uptake in Escherichia coli GN14176, a moderately norfloxacin-resistant clinical isolate, indicated that resistance was associated with both an altered drug target and a reduction in drug uptake.

  6. Preparation of Functional Soybean Protein Isolate

    Institute of Scientific and Technical Information of China (English)

    Liuzhitong; LiXiaolian; 等

    2000-01-01

    Soybean protein isolate(SPI) is a high purity soybean protein product.Its protein content is over 90%.A popular processing method is alkali dissolution and acid precipitation.This method can produce various functional SPIs by changing the temperature,pH,types of alkali and acid,and by different pretreatment and post transformation treatment.The properties addressed in this paper would open a big market for the appli cation of SPI.

  7. Preparation of Functional Soybean Protein Isolate

    Institute of Scientific and Technical Information of China (English)

    Liu Zhitong; Li Xiaolian; Zhao Guangming

    2000-01-01

    Soybean protein isolate(SPI)is a high purity soybean protein product. Its protein content is over 90% .A popular processing method is alkali dissolution and acid precipitation. This method can produce various functional SPIs by changing the temperature, pH, types of alkali and acid, and by different pretreatment and post transformation treatment. The properties addressed in this paper would open a big market for the appli cation of SPI.

  8. Drugs resistance and penicillinase activity in skin isolated Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Bhat K

    1990-01-01

    Full Text Available A study was carried out to evaluate the drug resistance pattern and penicillinase production in skin isolated Staphylococcus aurpus. The disk diffusion method showed prevalence of: multidrug resistance among S. aureus, strains, isolated from locafised skin abscesses. method for detection of penicilfinase could detect this enzyme m 98.60/o of the isolates all fo which were resistant to penicillin and ampicillin. C16xacillin resistance as detected by the agar dilution method was found in 1.4% of the isolates. On the whole cloxacillin and gentamy′cin were found to be the most effective ′antistaphylococcal antibotics.

  9. Contribution of β-lactamases and porin proteins OmpK35 and OmpK36 to carbapenem resistance in clinical isolates of KPC-2-producing Klebsiella pneumoniae.

    Science.gov (United States)

    Zhang, Ying; Jiang, Xiaofei; Wang, Yanyan; Li, Gang; Tian, Yueru; Liu, Hong; Ai, Fuqi; Ma, Yiming; Wang, Bei; Ruan, Feiyi; Rajakumar, Kumar

    2014-01-01

    Fifty-seven carbapenem-resistant Klebsiella pneumoniae isolates belonging to ST11 (50 isolates), ST423 (5 isolates), and two other sequence types were studied. All were positive for blaKPC-2, blaTEM-1, and blaCTX-M-14. SDS-PAGE analysis of six representative isolates demonstrated varied porin expression. Nevertheless, when blaKPC-2 was deleted, carbapenem resistance was markedly reduced. Additionally, SHV-12, DHA-1, and/or VIM-1 appeared to contribute to accessory carbapenemase activity. In contrast, OmpK35 and/or OmpK36 deficiency seemed to serve only as a minor cooperative factor.

  10. Fosfomycin susceptibility of canine methicillin-resistant Staphylococcus pseudintermedius isolates.

    Science.gov (United States)

    DiCicco, Matthew; Weese, Scott; Neethirajan, Suresh; Rousseau, Joyce; Singh, Ameet

    2014-04-01

    The effectiveness of fosfomycin was examined across 31 methicillin-resistant Staphylococcus pseudintermedius (MRSP) strains by agar dilution. Prevalence of the fosfomycin-resistance determinant gene, fosB, was assessed by PCR analysis. Results found that 84% of isolates were fosfomycin-susceptible. Interestingly, 87% of isolates possessed fosB, indicating no association between this putative staphylococci resistance gene and phenotypic resistance. Further evaluation of fosfomycin as a potential treatment of MRSP in dogs is warranted.

  11. Determining of antibiotic resistance profile inStaphylococcus aureus isolates

    Institute of Scientific and Technical Information of China (English)

    Hossein Motamedi; Hadis Mirzabeigi; Tahere Shirali

    2010-01-01

    Objective:To determine the pattern of antibiotic resistance amongStaphylococcus aureus (S. aureus) isolates from clinical specimens and to identify community-acquired methicillin-resistantStaphylococcus aureus(CA-MRSA)in specimens that have been collected from patients referring to one of the hospitals of Ahvaz.Methods:S. aureus isolates from a hospital in Ahvaz were screened for resistance to various antibiotics including methicillin. The susceptibility of the isolates was determined by Kirby-Bauer disc diffusion method. TheMRSA was also treated with ethidium bromide to find the origin of resistance.Results: Among the bacterial isolates, all of 11S. aureus were resistant to methicillin and cefixime,2 were resistant to ciprofloxacine,6 were resistant to tetracycline and the reminder were sensitive or intermediate to other antibiotics. The treated isolates were reminded resistant to methicillin and this suggested that the plasmid was not the origin of resistance in these isolates.Conclusions: These results showed that infection due toMRSA is widespread in Ahvaz and with respect to the spread of vancomycin resistance among MRSA and appearance of overwhelming infections. It is necessary to identify continuously the profile of antibiotic resistance amongS. aureus isolates in other regions and finding appropriate antibiotic for infection control and eradication.

  12. Genome-wide transcriptome analysis of fluoroquinolone resistance in clinical isolates of Escherichia coli.

    Science.gov (United States)

    Yamane, Takashi; Enokida, Hideki; Hayami, Hiroshi; Kawahara, Motoshi; Nakagawa, Masayuki

    2012-04-01

    Coincident with their worldwide use, resistance to fluoroquinolones in Escherichia coli has increased. To identify the gene expression profiles underlying fluoroquinolone resistance, we carried out genome-wide transcriptome analysis of fluoroquinolone-sensitive E. coli. Four fluoroquinolone-sensitive E. coli and five fluoroquinolone-resistant E. coli clinical isolates were subjected to complementary deoxyribonucleic acid microarray analysis. Some upregulated genes' expression was verified by real-time polymerase chain reaction using 104 E. coli clinical isolates, and minimum inhibitory concentration tests were carried out by using their transformants. A total of 40 genes were significantly upregulated in fluoroquinolone-resistant E. coli isolates (P fluoroquinolone-resistant E. coli. One of the phage shock protein operons, pspC, was significantly upregulated in 50 fluoroquinolone-resistant E. coli isolates (P fluoroquinolone-resistant E. coli. Deoxyribonucleic acid adenine methyltransferase (dam), which represses type I fimbriae genes, was significantly upregulated in the clinical fluoroquinolone-resistant E. coli isolates (P = 0.007). We established pspC- and dam-expressing E. coli transformants from fluoroquinolone-sensitive E. coli, and the minimum inhibitory concentration tests showed that the transformants acquired fluoroquinolone resistance, suggesting that upregulation of these genes contributes to acquiring fluoroquinolone resistance. Upregulation of psp operones and dam underlying pilus operons downregulation might be associated with fluoroquinolone resistance in E. coli. © 2011 The Japanese Urological Association.

  13. Amikacin resistance in Staphylococcus pseudintermedius isolated from dogs.

    Science.gov (United States)

    Gold, R M; Cohen, N D; Lawhon, S D

    2014-10-01

    Staphylococcus pseudintermedius is the most common microorganism isolated from canine pyoderma and postoperative wound infections. The prevalence of methicillin-resistant S. pseudintermedius (MRSP) has increased, and recently, isolates that are resistant not only to methicillin but also to other classes of antibiotic drugs, including aminoglycosides, have become common. A total of 422 S. pseudintermedius isolates collected from 413 dogs were analyzed for amikacin and methicillin resistance using broth microdilution and disk diffusion testing. Methicillin-resistant isolates were significantly (P Staphylococcus aureus, the most prevalent gene detected was aph(3')-IIIa found in 75% (24/32) of isolates followed by aac(6')/aph(2") and ant(4')-Ia in 12% (4/32) and 3% (1/32), respectively. Understanding the differences in antimicrobial resistance gene carriage between different species of Staphylococcus may improve antimicrobial drug selection for clinical therapy and provide insights into how resistance develops in S. pseudintermedius.

  14. Isolation of plant cell wall proteins.

    Science.gov (United States)

    Jamet, Elisabeth; Boudart, Georges; Borderies, Giséle; Charmont, Stephane; Lafitte, Claude; Rossignol, Michel; Canut, Herve; Pont-Lezica, Rafael

    2008-01-01

    The quality of a proteomic analysis of a cell compartment strongly depends on the reliability of the isolation procedure for the cell compartment of interest. Plant cell walls possess specific drawbacks: (1) the lack of a surrounding membrane may result in the loss of cell wall proteins (CWP) during the isolation procedure; (2) polysaccharide networks of cellulose, hemicelluloses, and pectins form potential traps for contaminants such as intracellular proteins; (3) the presence of proteins interacting in many different ways with the polysaccharide matrix require different procedures to elute them from the cell wall. Three categories of CWP are distinguished: labile proteins that have little or no interactions with cell wall components, weakly bound proteins extractable with salts, and strongly bound proteins. Two alternative protocols are decribed for cell wall proteomics: (1) nondestructive techniques allowing the extraction of labile or weakly bound CWP without damaging the plasma membrane; (2) destructive techniques to isolate cell walls from which weakly or strongly bound CWP can be extracted. These protocols give very low levels of contamination by intracellular proteins. Their application should lead to a realistic view of the cell wall proteome at least for labile and weakly bound CWP extractable by salts.

  15. Isolates of Staphylococcus aureus and saprophyticus resistant to antimicrobials isolated from the Lebanese aquatic environment.

    Science.gov (United States)

    Harakeh, Steve; Yassine, Hadi; Hajjar, Shady; El-Fadel, Mutasem

    2006-08-01

    The indiscriminate use of antimicrobials especially in developing countries has evoked serious bacterial resistance and led to the emergence of new and highly resistant strains of bacteria to commonly used antimicrobials. In Lebanon, pollution levels and bacterial infections are increasing at a high rate as a result of inadequate control measures to limit untreated effluent discharges into the sea or freshwater resources. The aim of this study was to isolate and molecularly characterize various Staphylococcus strains isolated from sea water, fresh water, sediments, and crab samples collected from representative communities along the coast of Lebanon. The results on the antimicrobial resistance indicated that the level of resistance of Staphylococcus aureus varied with various antimicrobials tested. The resistance patterns ranged between 45% in freshwater isolates and 54.8% in seawater ones. Fifty one percent of the tested isolates have shown resistance to at least one of the five tested antimicrobials; with seawater isolates exhibiting the highest rates of antimicrobial resistance.

  16. [Antioxidant activity of cationic whey protein isolate].

    Science.gov (United States)

    titova, M E; Komolov, S A; Tikhomirova, N A

    2012-01-01

    The process of lipid peroxidation (LPO) in biological membranes of cells is carried out by free radical mechanism, a feature of which is the interaction of radicals with other molecules. In this work we investigated the antioxidant activity of cationic whey protein isolate, obtained by the cation-exchange chromatography on KM-cellulose from raw cow's milk, in vitro and in vivo. In biological liquids, which are milk, blood serum, fetal fluids, contains a complex of biologically active substances with a unique multifunctional properties, and which are carrying out a protective, antimicrobial, regenerating, antioxidant, immunomodulatory, regulatory and others functions. Contents of the isolate were determined electrophoretically and by its biological activity. Cationic whey protein isolate included lactoperoxidase, lactoferrin, pancreatic RNase, lysozyme and angeogenin. The given isolate significantly has an antioxidant effect in model experimental systems in vitro and therefore may be considered as a factor that can adjust the intensity of lipid oxidation. In model solutions products of lipid oxidation were obtained by oxidation of phosphatidylcholine by hydrogen peroxide in the presence of a source of iron. The composition of the reaction mixture: 0,4 mM H2O2; 50 mcM of hemin; 2 mg/ml L-alpha-phosphatidylcholine from soybean (Sigma, German). Lipid peroxidation products were formed during the incubation of the reaction mixture for two hours at 37 degrees C. In our studies rats in the adaptation period immediately after isolation from the nest obtained from food given orally native cationic whey protein isolate at the concentration three times higher than in fresh cow's milk. On the manifestation of the antioxidant activity of cationic whey protein isolate in vivo evidence decrease of lipid peroxidation products concentration in the blood of rats from the experimental group receipt whey protein isolate in dos 0,6 mg/g for more than 20% (pwhey protein isolate has an

  17. Molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis clinical isolates

    Directory of Open Access Journals (Sweden)

    Meng Dong-Ya

    2014-01-01

    Full Text Available To evaluate the molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis (MH clinical strains isolated from urogenital specimens. 15 MH clinical isolates with different phenotypes of resistance to fluoroquinolones antibiotics were screened for mutations in the quinolone resistance-determining regions (QRDRs of DNA gyrase (gyrA and gyrB and topoisomerase IV (parC and parE in comparison with the reference strain PG21, which is susceptible to fluoroquinolones antibiotics. 15 MH isolates with three kinds of quinolone resistance phenotypes were obtained. Thirteen out of these quinolone-resistant isolates were found to carry nucleotide substitutions in either gyrA or parC. There were no alterations in gyrB and no mutations were found in the isolates with a phenotype of resistance to Ofloxacin (OFX, intermediate resistant to Levofloxacin (LVX and Sparfloxacin (SFX, and those susceptible to all three tested antibiotics. The molecular mechanism of fluoroquinolone resistance in clinical isolates of MH was reported in this study. The single amino acid mutation in ParC of MH may relate to the resistance to OFX and LVX and the high-level resistance to fluoroquinolones for MH is likely associated with mutations in both DNA gyrase and the ParC subunit of topoisomerase IV.

  18. Molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis clinical isolates.

    Science.gov (United States)

    Meng, Dong-Ya; Sun, Chang-Jian; Yu, Jing-Bo; Ma, Jun; Xue, Wen-Cheng

    2014-01-01

    To evaluate the molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis (MH) clinical strains isolated from urogenital specimens. 15 MH clinical isolates with different phenotypes of resistance to fluoroquinolones antibiotics were screened for mutations in the quinolone resistance-determining regions (QRDRs) of DNA gyrase (gyrA and gyrB) and topoisomerase IV (parC and parE) in comparison with the reference strain PG21, which is susceptible to fluoroquinolones antibiotics. 15 MH isolates with three kinds of quinolone resistance phenotypes were obtained. Thirteen out of these quinolone-resistant isolates were found to carry nucleotide substitutions in either gyrA or parC. There were no alterations in gyrB and no mutations were found in the isolates with a phenotype of resistance to Ofloxacin (OFX), intermediate resistant to Levofloxacin (LVX) and Sparfloxacin (SFX), and those susceptible to all three tested antibiotics. The molecular mechanism of fluoroquinolone resistance in clinical isolates of MH was reported in this study. The single amino acid mutation in ParC of MH may relate to the resistance to OFX and LVX and the high-level resistance to fluoroquinolones for MH is likely associated with mutations in both DNA gyrase and the ParC subunit of topoisomerase IV.

  19. Characterization of clarithromycin resistance in Malaysian isolates of Helicobacterpylori

    Institute of Scientific and Technical Information of China (English)

    Norazah Ahmad; Wan Rasinah Zakaria; Sheikh Anwar Abdullah; Ramelah Mohamed

    2009-01-01

    AIM: To characterize the types of mutations present in the 23S rRNA genes of Malaysian isolates of clarithromycin-resistant Helicobacter pylori (H pylori).METHODS: Clarithromycin susceptibility of H pylori isolates was determined by E test. Analyses for point mutations in the domain V of 23S rRNA genes in clarithromycin-resistant and -sensitive strains were performed by sequence analysis of amplified polymerase chain reaction products. Restriction fragment length polymorphism was performed using BsaⅠ and Mbo Ⅱ enzymes to detect restriction sites that correspond to the mutations in the clarithromycin-resistant strains.RESULTS: Of 187 isolates from 120 patients, four were resistant to clarithromycin, while 183 were sensitive. The MIC of the resistant strains ranged from 1.5 to 24 μg/mL. Two isolates had an A2142G mutation and another two had A2143G mutations. A T2182C mutation was detected in two out of four clarithromycin-resistant isolates and in 13 of 14 clarithromycin-sensitive isolates. Restriction enzyme analyses with Bsa I and Mbo n were able to detect the mutations.CONCLUSION: Clarithromycin resistance is an un-common occurrence among Malaysian isolates of Hpylori strains and the mutations A2142G and A2143G detected were associated with low-level resistance.

  20. Contribution of different mechanisms to the resistance to fluoroquinolones in clinical isolates of Salmonella enterica

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    Abeer Ahmed Rushdy

    Full Text Available OBJECTIVES: To study the potential factors include gene mutation, efflux pump and alteration of permeability associated with quinolone-resistance of Salmonella enterica strains isolated from patients with acute gastroenteritis and to evaluate the degree of synergistic activity of efflux pump inhibitors when combined with ciprofloxacin against resistant isolates. METHODS: Antimicrobial resistance patterns of fifty-eight Salmonella isolates were tested. Five isolates were selected to study the mechanism of resistance associated with quinolone group, including mutation in topoisomerase-encoding gene, altered cell permeability, and expression of an active efflux system. In addition, the combination between antibiotics and efflux pump inhibitors to overcome the microbial resistance was evaluated. RESULTS: Five Salmonella isolates totally resistant to all quinolones were studied. All isolates showed alterations in outer membrane proteins including disappearance of some or all of these proteins (Omp-A, Omp-C, Omp-D and Omp-F. Minimum inhibitory concentration values of ciprofloxacin were determined in the presence/absence of the efflux pump inhibitors: carbonyl cyanide m-chlorophenylhydrazone, norepinephrin and trimethoprim. Minimum inhibitory concentration values for two of the isolates were 2-4 fold lower with the addition of efflux pump inhibitors. All five Salmonella isolates were amplified for gyrA and parC genes and only two isolates were sequenced. S. Enteritidis 22 had double mutations at codon 83 and 87 in addition to three mutations at parC at codons 67, 76 and 80 whereas S. Typhimurium 57 had three mutations at codons 83, 87 and 119, but no mutations at parC. CONCLUSIONS: Efflux pump inhibitors may inhibit the major AcrAB-TolC in Salmonella efflux systems which are the major efflux pumps responsible for multidrug resistance in Gramnegative clinical isolates.

  1. Antimicrobial Resistance of Escherichia fergusonii Isolated from Broiler Chickens.

    Science.gov (United States)

    Simmons, Karen; Islam, M Rashedul; Rempel, Heidi; Block, Glenn; Topp, Edward; Diarra, Moussa S

    2016-06-01

    The objective of this study was to investigate the antibiotic resistance of Escherichia fergusonii isolated from commercial broiler chicken farms. A total of 245 isolates from cloacal and cecal samples of 28- to 36-day-old chickens were collected from 32 farms. Isolates were identified using PCR, and their susceptibility to 16 antibiotics was determined by disk diffusion assay. All isolates were susceptible to meropenem, amikacin, and ciprofloxacin. The most common resistances were against ampicillin (75.1%), streptomycin (62.9%), and tetracycline (57.1%). Of the 184 ampicillin-resistant isolates, 127 were investigated using a DNA microarray carrying 75 probes for antibiotic resistance genetic determinants. Of these 127 isolates, the β-lactamase blaCMY2, blaTEM, blaACT, blaSHV, and blaCTX-M-15 genes were detected in 120 (94.5%), 31 (24.4%), 8 (6.3%), 6 (4.7%), and 4 (3.2%) isolates, respectively. Other detected genes included those conferring resistance to aminoglycosides (aadA1, strA, strB), trimethoprims (dfrV, dfrA1), tetracyclines (tetA, tetB, tetC, tetE), and sulfonamides (sul1, sul2). Class 1 integron was found in 35 (27.6%) of the ampicillin-resistant isolates. However, our data showed that the tested E. fergusonii did not carry any carbapenemase blaOXA genes. Pulsed-field gel electrophoresis revealed that the selected ampicillin-resistant E. fergusonii isolates were genetically diverse. The present study indicates that the monitoring of antimicrobial-resistant bacteria should include enteric bacteria such as E. fergusonii, which could be a reservoir of antibiotic resistance genes. The detection of isolates harboring extended-spectrum β-lactamase genes, particularly blaCTX-M-15, in this work suggests that further investigations on the occurrence of such genes in broilers are warranted.

  2. Breast Cancer Resistance Protein Expression and 5-Fluorouracil Resistance

    Institute of Scientific and Technical Information of China (English)

    JIAN-HUI YUAN; ZHI-XIONG ZHUANG; JIN-QUAN CHENG; LONG-YUAN JIANG; WEI-DONG JI; LIANG-FENG GUO; JIAN-JUN LIU; XING-YUN XU; JING-SONG HE; XIAN-MING WANG

    2008-01-01

    To filtrate breast cancer resistance protein (BCRP)-mediated resistant agents and to investigate clinical relationship between BCRP expression and drug resistance. Methods MTT assay was performed to filtrate BCRP-mediated resistant agents with BCRP expression cell model and to detect chemosensitivity of breast cancer tissue specimens to these agents. A high performance liquid chromatography (HPLC) assay was established, and was used to measure the relative dose of intracellular retention resistant agents. RT-PCR and immununohistochemistry (IHC) were employed to investigate the BCRP expression in breast cancer tissue specimens. Results MTT assay showed that the expression of BCRP increased with the increasing resistance of 5-fluorouracil (5-Fu) (P=0.8124, P<0.01). Condusion Resistance to 5-Fu can be mediated by BCRP. Clinical chemotherapy for breast cancer patients can be optimized based on BCRP-positive expression.

  3. Small Molecule DFPM Derivative-Activated Plant Resistance Protein Signaling in Roots Is Unaffected by EDS1 Subcellular Targeting Signal and Chemical Genetic Isolation of victr R-Protein Mutants.

    Directory of Open Access Journals (Sweden)

    Hans-Henning Kunz

    Full Text Available The small molecule DFPM ([5-(3,4-dichlorophenylfuran-2-yl]-piperidine-1-ylmethanethione was recently shown to trigger signal transduction via early effector-triggered immunity signaling genes including EDS1 and PAD4 in Arabidopsis thaliana accession Col-0. Chemical genetic analyses of A. thaliana natural variants identified the plant Resistance protein-like Toll/Interleukin1 Receptor (TIR-Nucleotide Binding (NB-Leucine-Rich Repeat (LRR protein VICTR as required for DFPM-mediated root growth arrest. Here a chemical genetic screen for mutants which disrupt DFPM-mediated root growth arrest in the Col-0 accession identified new mutant alleles of the TIR-NB-LRR gene VICTR. One allele, victr-6, carries a Gly216-to-Asp mutation in the Walker A domain supporting an important function of the VICTR nucleotide binding domain in DFPM responses consistent with VICTR acting as a canonical Resistance protein. The essential nucleo-cytoplasmic regulator of TIR-NB-LRR-mediated effector-triggered immunity, EDS1, was reported to have both nuclear and cytoplasmic actions in pathogen resistance. DFPM was used to investigate the requirements for subcellular EDS1 localization in DFPM-mediated root growth arrest. EDS1-YFP fusions engineered to localize mainly in the cytoplasm or the nucleus by tagging with a nuclear export signal (NES or a nuclear localization signal (NLS, respectively, were tested. We found that wild-type EDS1-YFP and both the NES and NLS-tagged EDS1 variants were induced by DFPM treatments and fully complemented eds1 mutant plants in root responses to DFPM, suggesting that enrichment of EDS1 in either compartment could confer DFPM-mediated root growth arrest. We further found that a light and O2-dependent modification of DFPM is necessary to mediate DFPM signaling in roots. Chemical analyses including Liquid Chromatography-Mass Spectrometry and High-Resolution Atmospheric Pressure Chemical Ionization Mass Spectrometry identified a DFPM modification

  4. Small Molecule DFPM Derivative-Activated Plant Resistance Protein Signaling in Roots Is Unaffected by EDS1 Subcellular Targeting Signal and Chemical Genetic Isolation of victr R-Protein Mutants.

    Science.gov (United States)

    Kunz, Hans-Henning; Park, Jiyoung; Mevers, Emily; García, Ana V; Highhouse, Samantha; Gerwick, William H; Parker, Jane E; Schroeder, Julian I

    2016-01-01

    The small molecule DFPM ([5-(3,4-dichlorophenyl)furan-2-yl]-piperidine-1-ylmethanethione) was recently shown to trigger signal transduction via early effector-triggered immunity signaling genes including EDS1 and PAD4 in Arabidopsis thaliana accession Col-0. Chemical genetic analyses of A. thaliana natural variants identified the plant Resistance protein-like Toll/Interleukin1 Receptor (TIR)-Nucleotide Binding (NB)-Leucine-Rich Repeat (LRR) protein VICTR as required for DFPM-mediated root growth arrest. Here a chemical genetic screen for mutants which disrupt DFPM-mediated root growth arrest in the Col-0 accession identified new mutant alleles of the TIR-NB-LRR gene VICTR. One allele, victr-6, carries a Gly216-to-Asp mutation in the Walker A domain supporting an important function of the VICTR nucleotide binding domain in DFPM responses consistent with VICTR acting as a canonical Resistance protein. The essential nucleo-cytoplasmic regulator of TIR-NB-LRR-mediated effector-triggered immunity, EDS1, was reported to have both nuclear and cytoplasmic actions in pathogen resistance. DFPM was used to investigate the requirements for subcellular EDS1 localization in DFPM-mediated root growth arrest. EDS1-YFP fusions engineered to localize mainly in the cytoplasm or the nucleus by tagging with a nuclear export signal (NES) or a nuclear localization signal (NLS), respectively, were tested. We found that wild-type EDS1-YFP and both the NES and NLS-tagged EDS1 variants were induced by DFPM treatments and fully complemented eds1 mutant plants in root responses to DFPM, suggesting that enrichment of EDS1 in either compartment could confer DFPM-mediated root growth arrest. We further found that a light and O2-dependent modification of DFPM is necessary to mediate DFPM signaling in roots. Chemical analyses including Liquid Chromatography-Mass Spectrometry and High-Resolution Atmospheric Pressure Chemical Ionization Mass Spectrometry identified a DFPM modification product that is

  5. Biochemical evaluation of resistance responses of potato to different isolates of Alternaria solani.

    Science.gov (United States)

    Shahbazi, Hadis; Aminian, Heshmatollah; Sahebani, Navazollah; Halterman, Dennis A

    2010-05-01

    The resistance phenotypes of nine potato cultivars to five isolates of Alternaria solani, causal agent of early blight, were studied after inoculation and growth under greenhouse conditions. We identified potato cultivars with both susceptible and resistant phenotypes as well as A. solani isolates with varying degrees of aggressiveness. Two potato cultivars and two pathogen isolates were selected for biochemical analysis of phenol production and peroxidase activity after inoculation. Phenol compounds were evaluated 2, 4, 6, and 8 days after inoculation, while peroxidase activities were monitored daily for 10 days. Native polyacrylamide electrophoresis was used to identify one protein with peroxidase activity in extracts taken 6 days after inoculation. Significantly higher peroxidase activity as well as total phenol content in potato was correlated with resistance in the Iranian potato cultivar Diamond. Variability of responses within the same cultivar to different isolates of A. solani suggests genotypic diversity between isolates that results in phenotypic diversity for pathogen aggressiveness.

  6. Mechanisms of resistance to carbapenems in meropenem- resistant Acinetobacter isolates from clinical samples

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    Sinha M

    2007-01-01

    Full Text Available Purpose: To analyze the resistance mechanisms in Acinetobacter species by phenotypic methods. Methods: Antibiotic susceptibility profile for 150 clinical isolates of Acinetobacte r was determined by the standard disk diffusion method. Isolates detected to be meropenem resistant were tested further by broth microdilution minimum inhibitory concentration (MIC for meropenem. The resistant isolates were also tested for metallo β -lactamase (MBL production by the double-disk approximation test, for AmpC beta-lactamase production and efflux pump detection by agar microdilution MIC with and without reserpine. Results: Twenty-one isolates were found resistant to meropenem by the standard disk diffusion method. Nine samples were from patients admitted in intensive care units (ICUs. Broth microdilution MICs of the isolates revealed low-level resistance to meropenem. MBL was not produced by any of these isolates. AmpC β -lactamases were produced by nine (43% isolates. ′Efflux pump′-mediated resistance to meropenem was detected in two out of nine random isolates tested for the same . Conclusions: Carbapenem resistance is not uncommon in Acinetobacter isolates. AmpC production may cause carbapenem resistance. MBL and efflux pump may not be important causes of carbapenem resistance.

  7. Inducible clindamycin resistance among clinical isolates of staphylococci

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    Ciraj A

    2009-01-01

    Full Text Available Introduction: Clinical failure of clindamycin therapy has been reported due to multiple mechanisms that confer resistance to macrolide, lincosamide and streptogramin antibiotics. This study was undertaken to detect the presence of inducible clindamycin resistance among clinical isolates of staphylococci. Materials and Methods: The detection of inducible clindamycin resistance was performed by D-test using erythromycin and clindamycin discs as per CDC guidelines. Results: Among the 244 clinical isolates of staphylococci studied, 32 (13.1% showed inducible clindamycin resistance and belonged to the MLSBi phenotype. Among the MLS B i phenotypes, 10 isolates were methicillin-resistant Staphylococcus aureus (38.4% of the total MRSA, 16 were methicillin-sensitive Staphylococcus aureus (12.9% of the total MSSA and 6 were coagulase-negative staphylococci (6.3% of the total CONS. Conclusion: The test for inducible resistance to clindamycin should be included in the routine antibiotic susceptibility testing, as it will help in guiding therapy.

  8. Antibiotic resistances of intestinal lactobacilli isolated from wild boars.

    Science.gov (United States)

    Klose, Viviana; Bayer, Katharina; Kern, Corinna; Goelß, Florian; Fibi, Silvia; Wegl, Gertrude

    2014-01-10

    Acquired antibiotic resistances have been reported in lactobacilli of various animal and food sources, but there are no data from wild boar. The objective was a preliminary examination of the antibiotic resistance prevalence of intrinsically vancomycin-resistant lactobacilli isolated from wild boar intestines and analysis of the genetic determinants implicated. Out of three wild boars, 121 lactobacilli were recovered and grouped according to their whole cell protein patterns. Initial phenotypic screening revealed that all were susceptible to erythromycin (2 μg/ml), but 30 were resistant to tetracycline (32 μg/ml). Based on Randomly Amplified Polymorphic DNA-PCR clustering, 64 strains were selected as representative genotypes for identification and minimum inhibitory concentration (MIC) determination. Partial 16S rRNA gene sequencing identified four species: (i) L. mucosae (n=57), (ii) L. reuteri (n=47), (iii) L. fermentum (n=12), and (iv) L. murinus (n=5). Most heterofermentative strains displayed low MICs for ampicillin (AMP), chloramphenicol (CHL), streptomycin (STR), kanamycin (KAN), gentamicin (GEN), erythromycin (ERY), quinupristin/dalfopristin (Q/D), and clindamycin (CLI). Atypical MICs were found mainly in L. mucosae and L. reuteri for TET, KAN, STR, AMP and CHL, but except the TET MICs of L. mucosae mostly at low level. L. murinus strains revealed atypical MICs for aminoglycosides, and/or CHL, AMP, CLI. PCR screening detected tet(W) in 12 and tet(M) in one of heterofermentative strains, as well as the aph(3')-III kanamycin gene in L. murinus. This is the first report showing acquired antibiotic resistance determinants in intestinal lactobacilli of wild boar origin.

  9. Antimicrobial resistance in enterococci isolated from Turkey flocks fed virginiamycin.

    Science.gov (United States)

    Welton, L A; Thal, L A; Perri, M B; Donabedian, S; McMahon, J; Chow, J W; Zervos, M J

    1998-03-01

    From 125 separate cloacal cultures from three turkey flocks fed virginiamycin, 104 Enterococcus faecium and 186 Enterococcus faecalis isolates were obtained. As the turkeys aged, there was a higher percentage of quinupristin-dalfopristin-resistant E. faecium isolates, with isolates from the oldest flock being 100% resistant. There were no vancomycin-resistant enterococci. Results of pulsed-field gel electrophoresis (PFGE) indicated there were 11 PFGE types of E. faecalis and 7 PFGE types of E. faecium that were in more than one group of flock cultures.

  10. Microarray Evaluation of Antimicrobial Resistance and Virulence of Escherichia coli Isolates from Portuguese Poultry

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    Nuno Mendonça

    2016-01-01

    Full Text Available The presence of antimicrobial resistance and virulence factors of 174 Escherichia coli strains isolated from healthy Portuguese Gallus gallus was evaluated. Resistance profiles were determined against 33 antimicrobials by microbroth dilution. Resistance was prevalent for tetracycline (70% and ampicillin (63%. Extended-spectrum beta-lactamase (ESBL phenotype was observed in 18% of the isolates. Multidrug resistance was found in 56% of isolates. A subset of 74 isolates were screened by DNA microarrays for the carriage of 88 antibiotic resistance genes and 62 virulence genes. Overall, 37 different resistance genes were detected. The most common were tet(A (72%, blaTEM (68%, and sul1 (47%, while 21% isolates harbored an ESBL gene (blaCTX-M group 1, group 2, or group 9. Of these, 96% carried the increased serum survival (iss virulence gene, while 89% presented the enterobactin siderophore receptor protein (iroN, 70% the temperature-sensitive hemagglutinin (tsh, and 68% the long polar fimbriae (lpfA virulence genes associated with extraintestinal pathogenic E. coli. In conclusion, prevalence of antibiotic resistant E. coli from the microbiota of Portuguese chickens was high, including to extended spectrum cephalosporins. The majority of isolates seems to have the potential to trigger extraintestinal human infection due to the presence of some virulence genes. However, the absence of genes specific for enteropathogenic E. coli reduces the risk for human intestinal infection.

  11. Posttreatment Antifungal Resistance among Colonizing Candida Isolates in Candidemia Patients

    DEFF Research Database (Denmark)

    Jensen, R H; Johansen, H K; Søes, L M

    2015-01-01

    The prevalence of intrinsic and acquired resistance among colonizing Candida isolates from patients after candidemia was investigated systematically in a 1-year nationwide study. Patients were treated at the discretion of the treating physician. Oral swabs were obtained after treatment. Species...... analysis demonstrated a genetic relationship for 90% of all paired blood and oral isolates. Patients exposed to azoles for ≥ 7 days (n = 93) had a significantly larger proportion of species intrinsically less susceptible to azoles (particularly Candida glabrata) among oral isolates than among initial blood...... isolates (36.6% versus 12.9%; P 0.5). Acquired resistance in Candida albicans was rare (

  12. Import of proteins into isolated yeast mitochondria.

    Science.gov (United States)

    Peleh, Valentina; Ramesh, Ajay; Herrmann, Johannes M

    2015-01-01

    Mitochondria are essential organelles of eukaryotic cells. The vast majority of mitochondrial proteins is encoded within the nuclear genome and translocated into various mitochondrial compartments after translation in the cytosol as preproteins. Even in rather primitive eukaryotes like yeasts, there are 700-1,000 different proteins that need to be recognized in the cytosol, directed to the protein translocases in the two mitochondrial membranes and sorted to their appropriate mitochondrial subcompartment. In vitro reconstituted import systems have proved to be important tools to study these processes in detail. Using isolated mitochondria and radioactively labeled precursor proteins, it was possible to identify several import machineries and pathways consisting of a large number of components during the last few decades.

  13. Antibiotic resistance among Helicobacter pylori clinical isolates in Lima, Peru

    Science.gov (United States)

    Boehnke, Kevin F; Valdivieso, Manuel; Bussalleu, Alejandro; Sexton, Rachael; Thompson, Kathryn C; Osorio, Soledad; Reyes, Italo Novoa; Crowley, John J; Baker, Laurence H; Xi, Chuanwu

    2017-01-01

    Objectives Gastric carcinoma is the most common cancer and cause of cancer mortality in Peru. Helicobacter pylori, a bacterium that colonizes the human stomach, is a Group 1 carcinogen due to its causal relationship to gastric carcinoma. While eradication of H. pylori can help prevent gastric cancer, characterizing regional antibiotic resistance patterns is necessary to determine targeted treatment for each region. Thus, we examined primary antibiotic resistance in clinical isolates of H. pylori in Lima, Peru. Materials and methods H. pylori strains were isolated from gastric biopsies of patients with histologically proven H. pylori infection. Primary antibiotic resistance among isolates was examined using E-test strips. Isolates were examined for the presence of the cagA pathogenicity island and the vacA m1/m2 alleles via polymerase chain reaction. Results Seventy-six isolates were recovered from gastric biopsies. Clinical isolates showed evidence of antibiotic resistance to 1 (27.6%, n=21/76), 2 (28.9%, n=22/76), or ≥3 antibiotics (40.8%). Of 76 isolates, eight (10.5%) were resistant to amoxicillin and clarithromycin, which are part of the standard triple therapy for H. pylori infection. No trends were seen between the presence of cagA, vacA m1, or vacA m2 and antibiotic resistance. Conclusion The rate of antibiotic resistance among H. pylori isolates in Lima, Peru, is higher than expected and presents cause for concern. To develop more targeted eradication therapies for H. pylori in Peru, more research is needed to better characterize antibiotic resistance among a larger number of clinical isolates prospectively. PMID:28331349

  14. ANTIMICROBIAL RESISTANCE PATTERN OF STAPHYLOCOCCUS AUREUS ISOLATES FROM DAKSHINA KANNADA

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    Rao Venkatakrishna

    2011-03-01

    Full Text Available Methicillin resistant Staphylococcus aureus (MRSA is an important cause of infections in hospitals and pose a great challenge to the treating clinicians; even emergence of vancomycin resistance has been reported. Therefore the knowledge of prevalence of MRSA and their antimicrobial profile becomes necessary. This study is aimed to determine prevalence of MRSA and their antimicrobial sensitivity pattern in Dakshina Kannada.Clinical specimens and carrier samples were cultured as per standard methods. The isolates were identified by using catalase test, coagulase tube test, mannitol fermentation and DNAase test. Antimicrobial susceptibility test was done for the isolates as per Kirby-Bauer disc diffusion method; the isolates were also tested for methicillin resistance using oxacillin and cefoxitin discs.A total of 250 isolates were tested (200 clinical isolates and 50 from carriers and 67 MRSA isolates were obtained (52 clinical samples and 15 from carriers. The degree of resistance to penicillin, ampicillin, ciprofloxacin, clindamycin and erythromycin were 100%, 100%, 53-56%, 14-16 % and 45-48% respectively. Resistance to vancomycin was not found. As the degree of resistance of MRSA towards antibiotics varies from region to region, in vitro susceptibility testing of every isolate of MRSA in clinical laboratories is inevitable.

  15. Changing patterns of drug-resistant Shigella isolates in egypt.

    Science.gov (United States)

    Abd-Elmeged, Ghada M; Khairy, Rasha M; Abo-Eloyoon, Sahar M; Abdelwahab, Sayed F

    2015-06-01

    The emergence of multidrug resistance (MDR) is a serious problem in treating shigellosis. There are limited existing data examining the change in the antimicrobial resistance profile of Shigella in Egypt. We previously reported that 58% of the Shigella isolates in Egypt were resistant to at least one member of the three different antimicrobial groups. This study was performed to determine the antimicrobial resistance profile of Shigella, determine their possible mechanisms of resistance, and compare their resistance profile to those reported 20 years ago. Stool samples were collected from 500 subjects and processed for the isolation and identification of Shigella. The susceptibility of the isolates to 11 different antimicrobials was determined using the disc diffusion method. Of 500 stool cultures, 24 (4.8%) samples were positive for Shigella. There was a high percentage of resistance to ampicillin (88%), tetracycline (83%), and sulfamethoxazole-trimethoprim (75%). Also, there was a moderate percentage of resistance to chloramphenicol (46%), streptomycin (42%), ceftazidime (33%), and cefotaxime (25%). A lower percentage of resistance was recorded for amikacin, nalidixic acid (17% each), and ofloxacin (7%), while no resistance was found to ciprofloxacin (0%). Twenty-one of the isolates (88%) were resistant to at least three different antimicrobial groups (indicating MDR). The average number of antimicrobial agents to which the Shigella isolates were resistant was 4.3±1.4, while it was 3.4±1.5 in the same locality in 1994. These data demonstrate that there is a marked increase in MDR and change in the resistance patterns of Shigella over the past 20 years.

  16. Soy protein isolate molecular level contributions to bulk adhesive properties

    Science.gov (United States)

    Shera, Jeanne Norton

    Increasing environmental awareness and the recognized health hazards of formaldehyde-based resins has prompted a strong demand for environmentally-responsible adhesives for wood composites. Soy protein-based adhesives have been shown to be commercially viable with 90-day shelf stability and composite physical properties comparable to those of commercial formaldehyde-based particleboards. The main research focus is to isolate and characterize the molecular level features in soy protein isolate responsible for providing mechanical properties, storage stability, and water resistance during adhesive formulation, processing, and wood composite fabrication. Commercial composite board will be reviewed to enhance our understanding of the individual components and processes required for particleboard production. The levels of protein structure will be defined and an overview of current bio-based technology will be presented. In the process, the logic for utilizing soy protein as a sole binder in the adhesive will be reinforced. Variables such as adhesive components, pH, divalent ions, blend aging, protein molecular weight, formulation solids content, and soy protein functionalization will relate the bulk properties of soy protein adhesives to the molecular configuration of the soybean protein. This work has demonstrated that when intermolecular beta-sheet interactions and protein long-range order is disrupted, viscosity and mechanical properties decrease. Storage stability can be maintained through the stabilization of intermolecular beta-sheet interactions. When molecular weight is reduced through enzymatic digestion, long-range order is disrupted and viscosity and mechanical properties decrease accordingly. Processibility and physical properties must be balanced to increase solids while maintaining low viscosity, desirable mechanical properties, and adequate storage stability. The structure of the soybean protein must be related to the particleboard bulk mechanical

  17. Genomic analyses of DNA transformation and penicillin resistance in Streptococcus pneumoniae clinical isolates.

    Science.gov (United States)

    Fani, Fereshteh; Leprohon, Philippe; Zhanel, George G; Bergeron, Michel G; Ouellette, Marc

    2014-01-01

    Alterations in penicillin-binding proteins, the target enzymes for β-lactam antibiotics, are recognized as primary penicillin resistance mechanisms in Streptococcus pneumoniae. Few studies have analyzed penicillin resistance at the genome scale, however, and we report the sequencing of S. pneumoniae R6 transformants generated while reconstructing the penicillin resistance phenotypes from three penicillin-resistant clinical isolates by serial genome transformation. The genome sequences of the three last-level transformants T2-18209, T5-1983, and T3-55938 revealed that 16.2 kb, 82.7 kb, and 137.2 kb of their genomes had been replaced with 5, 20, and 37 recombinant sequence segments derived from their respective parental clinical isolates, documenting the extent of DNA transformation between strains. A role in penicillin resistance was confirmed for some of the mutations identified in the transformants. Several multiple recombination events were also found to have happened at single loci coding for penicillin-binding proteins (PBPs) that increase resistance. Sequencing of the transformants with MICs for penicillin similar to those of the parent clinical strains confirmed the importance of mosaic PBP2x, -2b, and -1a as a driving force in penicillin resistance. A role in resistance for mosaic PBP2a was also observed for two of the resistant clinical isolates.

  18. Cytosolic Proteome Profiling of Aminoglycosides Resistant Mycobacterium tuberculosis Clinical Isolates Using MALDI-TOF/MS

    Science.gov (United States)

    Sharma, Divakar; Lata, Manju; Singh, Rananjay; Deo, Nirmala; Venkatesan, Krishnamurthy; Bisht, Deepa

    2016-01-01

    Emergence of extensively drug resistant tuberculosis (XDR-TB) is the consequence of the failure of second line TB treatment. Aminoglycosides are the important second line anti-TB drugs used to treat the multi drug resistant tuberculosis (MDR-TB). Main known mechanism of action of aminoglycosides is to inhibit the protein synthesis by inhibiting the normal functioning of ribosome. Primary target of aminoglycosides are the ribosomal RNA and its associated proteins. Various mechanisms have been proposed for aminoglycosides resistance but still some are unsolved. As proteins are involved in most of the biological processes, these act as a potential diagnostic markers and drug targets. In the present study we analyzed the purely cytosolic proteome of amikacin (AK) and kanamycin (KM) resistant Mycobacterium tuberculosis isolates by proteomic and bioinformatic approaches. Twenty protein spots were found to have over expressed in resistant isolates and were identified. Among these Rv3208A, Rv2623, Rv1360, Rv2140c, Rv1636, and Rv2185c are six proteins with unknown functions or undefined role. Docking results showed that AK and KM binds to the conserved domain (DUF, USP-A, Luciferase, PEBP and Polyketidecyclase/dehydrase domain) of these hypothetical proteins and over expression of these proteins might neutralize/modulate the effect of drug molecules. TBPred and GPS-PUP predicted cytoplasmic nature and potential pupylation sites within these identified proteins, respectively. String analysis also suggested that over expressed proteins along with their interactive partners might be involved in aminoglycosides resistance. Cumulative effect of these over expressed proteins could be involved in AK and KM resistance by mitigating the toxicity, repression of drug target and neutralizing affect. These findings need further exploitation for the expansion of newer therapeutics or diagnostic markers against AK and KM resistance so that an extreme condition like XDR-TB can be prevented

  19. Antimicrobial resistance patterns in Danish isolates of Flavobacterium psychrophilum

    DEFF Research Database (Denmark)

    Bruun, Morten Sichlau; Schmidt, A.S.; Madsen, Lone

    2000-01-01

    were tested and the resulting antibiograms were used to predict the theoretical therapeutic efficacy and to evaluate if resistance had changed as a course of time. Antimicrobial agents included in this investigation were oxolinic acid (OXA), amoxicillin (AMX), potentiated sulfadiazine, oxytetracycline......The resistance pattern of Flavobacterium psychrophilum to the antimicrobial agents used in fish farming in Denmark was assessed in vitro using an agar dilution method. After identification of 387 isolates from clinical outbreaks of rainbow trout fry syndrome (RTFS) and the environment, the isolates...... (OTC) and florfenicol (FLO). We found that F. psychrophilum isolates divided in susceptible and resistant clusters reflecting the reduced efficacy in practice when using OTC and AMX. The most recent isolates were less susceptible to AMX and OXA, whereas resistance to OTC seemed stable over the last 5...

  20. Isolation and characterization of multidrug-resistant side population ...

    African Journals Online (AJOL)

    Isolation and characterization of multidrug-resistant side population cells in prostate carcinoma. ... PROMOTING ACCESS TO AFRICAN RESEARCH ... Keywords: Side population cells, ABC transporters, Cancer stem cells, Chemotherapy, ...

  1. Molecular Detection of Inducible Clindamycin Resistance among Staphylococcal Strains Isolated from Hospital Patients

    Directory of Open Access Journals (Sweden)

    Shadiyeh Abdollahi

    2013-04-01

    Full Text Available Background & Objectives: Macrolide, lincosamide and streptogramin B (MLSB antimicrobial agents are used in the treatment of staphylococcal infections. They prevent the microbial protein synthesis system through binding to 23 S rRNA. The aim of this study was to apply molecular methods to detect inducible clindamycin resistance genes among staphylococcal strains isolated from clinical specimens.   Methods : Two hundred staphylococcus strains were isolated from nose and throat swabs of patients in Toohid and Besat hospitals in Sanandaj . Antimicrobial susceptibilities of isolates were determined using disc diffusion method, agar screen test and D-Test. A multiplex PCR was performed using primers specific for erm (A, B, C, TR genes.   Results: Out of 200 isolates, 18.5 % were MRSA and 32% were MRCNS (methicillin resistant coagulase negative staphylococci. Of 80 erythromycin resistant isolates, 48 were coagulase negative and 32 were S. aureus. Among the 48 coagulase negative staphylococci (CONS isolates, 11.63% expressed the MLSB-inducible phenotypes. Using PCR, the frequency of different genes in the collection of isolates were as follows: ermA 5.41 % , erm B 5.41 % , and erm C 3.13%. The ermTR gene was negative in all isolates. Among the 32 S. aureus isolates, 9.38% expressed the MLSB-nducible phenotype. Using PCR, these isolates harbored erm A (2.22%, ermB (2.22%, ermC (2.22% and ermTR (2.22% .   Conclusion: This is the first study to show the rate of inducible clindamycin clinical isolates of staphylococci harboring erm genes in Sananadaj. It also demonstrated the frequency of erm genes was higher among CONS isolates than S. aureus. This data suggested the transfer of resistance gene from nonpathogenic to pathogenic strains is likely to happen. Therefore, screening and control of these resistance genes is recommended at clinical laboratories.

  2. Molecular characterization, fitness and mycotoxin production of benzimidazole-resistant isolates of Penicillium expansum.

    Science.gov (United States)

    Malandrakis, Anastasios A; Markoglou, Anastasios N; Konstantinou, Sotiris; Doukas, Eleftherios G; Kalampokis, John F; Karaoglanidis, George S

    2013-04-01

    Penicillium expansum field-strains resistant to benzimidazole fungicides were isolated in high frequency from decayed apple fruit collected from packinghouses and processing industries located in the region of Imathia, N. Greece. In vitro fungitoxicity tests resulted in the identification of two different resistant phenotypes: highly (BEN-HR) and moderately (BEN-MR) carbendazim-resistant. Thirty seven percent of the isolated P. expansum strains belonged to the BEN-HR phenotype, carried no apparent fitness penalties and exhibited resistance levels higher than 60 based on EC50 values. Cross resistance studies with other benzimidazole fungicides showed that all BEN-HR and BEN-MR isolates were also less sensitive to benomyl and thiabendazole. Fungitoxicity tests on the response of BEN-HR isolates to fungicides belonging to other chemical classes revealed no cross-resistance relationships between benzimidazoles and the phenylpyrrole fludioxonil, the dicarboximide iprodione, the anilinopyrimidine cyprodinil, the QoI pyraclostrobin, the imidazole imazalil and the triazole tebuconazole, indicating that a target-site modification is probably responsible for the BEN-HR phenotype observed. Contrary to the above, some BEN-MR isolates exhibited an increased sensitivity to cyprodinil compared to benzimidazole-sensitive ones. BEN-MR isolates had fitness parameters similar to the benzimidazole-sensitive isolates except for conidia production which appeared significantly decreased. Analysis of mycotoxin production (patulin and citrinin) showed that all benzimidazole-resistant isolates produced mycotoxins at concentrations significantly higher than sensitive isolates both on culture medium and on artificially inoculated apple fruit. Comparison of the β-tubulin gene DNA sequence between resistant and sensitive isolates revealed a point mutation resulting from the E198A substitution of the corresponding protein in most but not all HR isolates tested. Molecular analysis of the

  3. DIRProt: a computational approach for discriminating insecticide resistant proteins from non-resistant proteins.

    Science.gov (United States)

    Meher, Prabina Kumar; Sahu, Tanmaya Kumar; Banchariya, Anjali; Rao, Atmakuri Ramakrishna

    2017-03-24

    Insecticide resistance is a major challenge for the control program of insect pests in the fields of crop protection, human and animal health etc. Resistance to different insecticides is conferred by the proteins encoded from certain class of genes of the insects. To distinguish the insecticide resistant proteins from non-resistant proteins, no computational tool is available till date. Thus, development of such a computational tool will be helpful in predicting the insecticide resistant proteins, which can be targeted for developing appropriate insecticides. Five different sets of feature viz., amino acid composition (AAC), di-peptide composition (DPC), pseudo amino acid composition (PAAC), composition-transition-distribution (CTD) and auto-correlation function (ACF) were used to map the protein sequences into numeric feature vectors. The encoded numeric vectors were then used as input in support vector machine (SVM) for classification of insecticide resistant and non-resistant proteins. Higher accuracies were obtained under RBF kernel than that of other kernels. Further, accuracies were observed to be higher for DPC feature set as compared to others. The proposed approach achieved an overall accuracy of >90% in discriminating resistant from non-resistant proteins. Further, the two classes of resistant proteins i.e., detoxification-based and target-based were discriminated from non-resistant proteins with >95% accuracy. Besides, >95% accuracy was also observed for discrimination of proteins involved in detoxification- and target-based resistance mechanisms. The proposed approach not only outperformed Blastp, PSI-Blast and Delta-Blast algorithms, but also achieved >92% accuracy while assessed using an independent dataset of 75 insecticide resistant proteins. This paper presents the first computational approach for discriminating the insecticide resistant proteins from non-resistant proteins. Based on the proposed approach, an online prediction server DIRProt has

  4. Analysis of natural isolates of Lactobacilli resistant to bacteriocin nisin

    Directory of Open Access Journals (Sweden)

    Strahinić Ivana D.

    2005-01-01

    Full Text Available The collection of lactic acid bacteria (LAB was made by isolation of microorganisms from fermented products traditionally manufactured in different geographical regions (high mountains, river valleys, seaside, etc. Among collected LAB, 51 isolates were identified as Lactobacillus sp. Results showed that all isolated lactobacilli were mesophilic strains, since they grew at 15°C and 30°C but not at 45°C. Testing the ability of isolated lactobacilli to grow in the presence of nisin revealed that Lactobacillus sp. isolates designed BGCGK4, BGHN40, BGBUK2-8, BGBUK2-7 and BGBUK2-16 were resistant to nisin. Determination of the minimal inhibitory concentrations (MIC for nisin revealed that the most resistant isolate was Lactobacillus sp. BGCGK4. Isolate BGBUK2-16, determined as Lactobacillus paracasei subsp. paracasei, produces bacteriocin, named Bac217 and showed a resistance to 8000 IU/ml of nisin. Plasmid curing of BGBUK2-16 resulted in derivatives BGBUK2-16/K2 and BGBUK2-16/K4. Derivative BGBUK2-16/K2 retained resistance to Bac217 and nisin, but lost the ability to synthesise Bac217. Derivative BGBUK2-16/K4 lost concomitantly the resistance to both Bac217 and nisin.

  5. Antimicrobial resistance of Salmonella spp. isolated from food

    Science.gov (United States)

    Mąka, Łukasz; Popowska, Magdalena

    This review summarizes current data on resistance among Salmonella spp. isolates of food origin from countries in different regions of the world. The mechanisms of resistance to different groups of antimicrobial compounds are also considered. Among strains resistant to quinolones and/or fluoroquinolones the most prevalent mechanism is amino acid substitutions in quinolone resistance-determining region (QRDR) of genes gyrA, parC but mechanism of growing importance is plasmid-mediated quinolone resistance (PMQR) associated with genes qnrA, qnrB, qnrC, qnrD, qnrS but frequency of their detection is different. Resistance to sulfonamides is mostly associated with genes sul1 and sul2, while resistance to trimethoprim is associated with various variants of dhfr ( dfr) genes. Taking into account Salmonella spp. strains isolated from food, resistance to β-lactams is commonly associated with β-lactamases encoding by blaTEM genes. However strains ESBL and AmpC – positive are also detected. Resistance to aminoglicosides is commonly result of enzymatic inactivation. Three types of aminoglycoside modifying enzyme are: acetyltransferases (AAC), adenyltransferases (ANT) and phosphotransferases (APH). Resistance to tetracyclines among Salmonella spp. isolated from food is most commonly associated with active efflux. Among numerous genetic determinants encoding efflux pumps tetA, tetB, tetC, tetD, tetE and tetG are reported predominatingly. One of the most common mechanisms of resistance against chloramphenicol is its inactivation by chloramphenicol acetyltrasferases (CATs), but resistance to this compound can be also mediated by chloramphenicol efflux pumps encoded by the genes cmlA and floR. It is important to monitor resistance of Salmonella isolated from food, because the globalization of trade, leading to the long-distance

  6. Antimicrobial Susceptibility and Characterization of Outer Membrane Proteins of Aeromonas hydrophila Isolated in China

    Institute of Scientific and Technical Information of China (English)

    GUO Peng; WANG Na; LIU Yong-jie; LU Cheng-ping

    2014-01-01

    Aeromonas hydrophila isolates from clinical cases (n=43) were tested against 8 antimicrobial agents and typed by outer membrane protein (OMP) pattern by using sodium dodecyl sulfate gel electrophoresis. All isolates were resistant to ampicillin (MICs, 16 mg mL-1) and sulfamonomethoxine (MICs, 64 mg mL-1), but susceptible to norlfoxacin (MICs, 0.5 mg mL-1). There was a high incidence of resistance to erythromycin (90.70%) and tylosin (93.02%), while a low incidences of resistance to ciprofloxacin (2.33%), enrofloxacin (2.33%) and florfenicol (4.65%). Six different outer membrane protein patterns were found among 34 isolates by analyzing proteins in the range of 22 to 50 kDa, other than 9 isolates with their respective proifles. The strains with the similar OMP proifles had similar resistances. Compared with the other strains from the same OMP patterns, NB-1, A.Pun and MR-1 had lacked the proteins in the range of 30 to 45 kDa and their resistance to lforfenicol substantially increased. It is speculated that the outer membrane protein changes might correlate with decreased susceptibility to lforfenicol in the three strains. Some strains which showed completely identical OMP types had a little difference in their resistance to lfuoroquinolones, indicating that there might be other factors that were involved in the antimicrobial resistance of A. hydrophila.

  7. AMINOGLYCOSIDE RESISTANCE GENES IN Pseudomonas aeruginosa ISOLATES FROM CUMANA, VENEZUELA.

    Science.gov (United States)

    Teixeira, Bertinellys; Rodulfo, Hectorina; Carreño, Numirin; Guzmán, Militza; Salazar, Elsa; De Donato, Marcos

    2016-01-01

    The enzymatic modification of aminoglycosides by aminoglycoside-acetyltransferases (AAC), aminoglycoside-adenyltransferases (AAD), and aminoglycoside-phosphotransferases (APH), is the most common resistance mechanism in P. aeruginosa and these enzymes can be coded on mobile genetic elements that contribute to their dispersion. One hundred and thirty seven P. aeruginosa isolates from the University Hospital, Cumana, Venezuela (HUAPA) were evaluated. Antimicrobial susceptibility was determined by the disk diffusion method and theaac, aadB and aph genes were detected by PCR. Most of the P. aeruginosa isolates (33/137) were identified from the Intensive Care Unit (ICU), mainly from discharges (96/137). The frequency of resistant P. aeruginosaisolates was found to be higher for the aminoglycosides tobramycin and amikacin (30.7 and 29.9%, respectively). Phenotype VI, resistant to these antibiotics, was the most frequent (14/49), followed by phenotype I, resistant to all the aminoglycosides tested (12/49). The aac(6´)-Ib,aphA1 and aadB genes were the most frequently detected, and the simultaneous presence of several resistance genes in the same isolate was demonstrated. Aminoglycoside resistance in isolates ofP. aeruginosa at the HUAPA is partly due to the presence of the aac(6´)-Ib, aphA1 andaadB genes, but the high rates of antimicrobial resistance suggest the existence of several mechanisms acting together. This is the first report of aminoglycoside resistance genes in Venezuela and one of the few in Latin America.

  8. Posttreatment Antifungal Resistance among Colonizing Candida Isolates in Candidemia Patients

    DEFF Research Database (Denmark)

    Jensen, R H; Johansen, H K; Søes, L M

    2016-01-01

    The prevalence of intrinsic and acquired resistance among colonizing Candida isolates from patients after candidemia was investigated systematically in a 1-year nationwide study. Patients were treated at the discretion of the treating physician. Oral swabs were obtained after treatment. Species......, especially C. glabrata, following treatment for candidemia. The resistance rates were high, raising concern in general for patients exposed to antifungal drugs....

  9. AMINOGLYCOSIDE RESISTANCE GENES IN Pseudomonas aeruginosa ISOLATES FROM CUMANA, VENEZUELA

    Science.gov (United States)

    TEIXEIRA, Bertinellys; RODULFO, Hectorina; CARREÑO, Numirin; GUZMÁN, Militza; SALAZAR, Elsa; DONATO, Marcos DE

    2016-01-01

    The enzymatic modification of aminoglycosides by aminoglycoside-acetyltransferases (AAC), aminoglycoside-adenyltransferases (AAD), and aminoglycoside-phosphotransferases (APH), is the most common resistance mechanism in P. aeruginosa and these enzymes can be coded on mobile genetic elements that contribute to their dispersion. One hundred and thirty seven P. aeruginosa isolates from the University Hospital, Cumana, Venezuela (HUAPA) were evaluated. Antimicrobial susceptibility was determined by the disk diffusion method and theaac, aadB and aph genes were detected by PCR. Most of the P. aeruginosa isolates (33/137) were identified from the Intensive Care Unit (ICU), mainly from discharges (96/137). The frequency of resistant P. aeruginosaisolates was found to be higher for the aminoglycosides tobramycin and amikacin (30.7 and 29.9%, respectively). Phenotype VI, resistant to these antibiotics, was the most frequent (14/49), followed by phenotype I, resistant to all the aminoglycosides tested (12/49). The aac(6´)-Ib,aphA1 and aadB genes were the most frequently detected, and the simultaneous presence of several resistance genes in the same isolate was demonstrated. Aminoglycoside resistance in isolates ofP. aeruginosa at the HUAPA is partly due to the presence of the aac(6´)-Ib, aphA1 andaadB genes, but the high rates of antimicrobial resistance suggest the existence of several mechanisms acting together. This is the first report of aminoglycoside resistance genes in Venezuela and one of the few in Latin America. PMID:27007556

  10. Norfloxacin resistance in a clinical isolate of Escherichia coli.

    Science.gov (United States)

    Aoyama, H; Sato, K; Kato, T; Hirai, K; Mitsuhashi, S

    1987-01-01

    Analysis of DNA gyrase supercoiling and of norfloxacin uptake in Escherichia coli GN14176, a moderately norfloxacin-resistant clinical isolate, indicated that resistance was associated with both an altered drug target and a reduction in drug uptake. Images PMID:2829712

  11. Drug efflux proteins in multidrug resistant bacteria

    NARCIS (Netherlands)

    vanVeen, HW; Konings, WN

    1997-01-01

    Bacteria contain an array of transport proteins in their cytoplasmic membrane. Many of these proteins play an important role in conferring resistance to toxic compounds. The multidrug efflux systems encountered in prokaryotic cells are very similar to those observed in eukaryotic cells. Therefore, a

  12. Drug efflux proteins in multidrug resistant bacteria

    NARCIS (Netherlands)

    vanVeen, HW; Konings, WN

    Bacteria contain an array of transport proteins in their cytoplasmic membrane. Many of these proteins play an important role in conferring resistance to toxic compounds. The multidrug efflux systems encountered in prokaryotic cells are very similar to those observed in eukaryotic cells. Therefore, a

  13. Selection of tomato plants resistant to a local Polish isolate of tomato spotted wilt virus (TSWV).

    Science.gov (United States)

    Czech, Andrzej S; Szklarczyk, Marek; Gajewski, Zbigniew; Zukowska, Ewa; Michalik, Barbara; Kobyłko, Tadeusz; Strzałka, Kazimierz

    2003-01-01

    We found that the Sw-5 gene confers resistance to one of the Polish isolates of tomato spotted wilt virus (TSWV). A series of tomato breeding accessions was analysed along with standards of resistance and susceptibility to TSWV. The presence of the Sw-5 gene was determined using the available PCR marker. Subsequently plants from these accessions were grown in the presence of the TSWV isolate from Poland. Some of them developed severe symptoms of the TSWV disease. Expression of the virus proteins was also assayed in tissues of the investigated plants. We found general agreement between either lack or presence of the disease symptoms, virus proteins and resistance gene. Some observed discrepancies of these data are also discussed. Our results indicate that marker-assisted selection can be used for breeding of the TSWV-resistant tomato in Poland.

  14. Multiple antibiotic resistance among gram negative bacteria isolated from poultry.

    Science.gov (United States)

    Ansari, F A; Khatoon, H

    1994-03-01

    Gram negative bacteria, including species of Salmonella, Escherichia, Pseudomonas and Klebsiella, isolated from poultry, were screened for their resistance to the commonly used antibiotics: ampicillin, chloramphenicol, gentamycin, kanamycin, neomycin, polymyxin B, streptomycin and tetracycline. Of the 500 bacteria screened, 351 were found to be resistant to one or more antibiotics at the level of 50 micrograms/ml. Various patterns of antibiotic resistance observed during these studies have been reported.

  15. Comparative proteomics study on meglumine antimoniate sensitive and resistant Leishmania tropica isolated from Iranian anthroponotic cutaneous leishmaniasis patients.

    Science.gov (United States)

    Hajjaran, H; Azarian, B; Mohebali, M; Hadighi, R; Assareh, A; Vaziri, B

    2012-02-01

    In order to define the protein expressional changes related to the process of meglumine antimoniate resistance in anthroponotic cutaneous leishmaniasis (CL), we performed a comparative proteomics analysis on sensitive and resistant strains of Leishmania tropica isolated from Iranian CL patients. Cell proteins were analysed with 2-dimensional electrophoresis and differentially expressed proteins were identified by matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry. Image analysis of the matched maps identified 7 proteins that were either over- or down-expressed: activated protein kinase c receptor(LACK), alpha tubulin (x2), prostaglandin f2-alpha synthase, protein disulfide isomerase, vesicular transport protein and a hypothetical protein. The study shows the usefulness of proteomics in identifying proteins that may express differences between sensitive and resistant L. tropica isolates.

  16. Antimicrobial resistance in Enterococcus strains isolated from healthy domestic dogs.

    Science.gov (United States)

    Bertelloni, Fabrizio; Salvadori, Claudia; Lotti, Giulia; Cerri, Domenico; Ebani, Valentina Virginia

    2016-12-15

    Enterococci are opportunistic bacteria that cause severe infections in animals and humans, capable to acquire, express, and transfer antimicrobial resistance. Susceptibility to 21 antimicrobial agents was tested by the disk diffusion method in 222 Enterococcus spp. strains isolated from the fecal samples of 287 healthy domestic dogs. Vancomycin and ampicillin minimum inhibitory concentrations (MICs) and high-level aminoglycoside resistance (HLAR) tests were also performed. Isolates showed resistance mainly to streptomycin (88.7%), neomycin (80.6%), and tetracycline (69.4%). Forty-two (18.9%) isolates showed an HLAR to streptomycin and 15 (6.7%) to gentamicin. Vancomycin and ampicillin MIC values showed 1 and 18 resistant strains, respectively. One hundred and thirty-six (61.2%) strains were classified as multidrug resistant and six (2.7%) strains as possibly extensively drug-resistant bacteria. Enterococcus faecium and Enterococcus faecalis were the most prevalent antimicrobial resistant species. Companion animals, which often live in close contact with their owners and share the same environment, represent a serious source of enterococci resistant to several antibiotics; for this reason, they may be a hazard for public health by providing a conduit for the entrance of resistance genes into the community.

  17. Echinocandin Resistance in Candida Species Isolates from Liver Transplant Recipients.

    Science.gov (United States)

    Prigent, Gwénolé; Aït-Ammar, Nawel; Levesque, Eric; Fekkar, Arnaud; Costa, Jean-Marc; El Anbassi, Sarra; Foulet, Françoise; Duvoux, Christophe; Merle, Jean-Claude; Dannaoui, Eric; Botterel, Françoise

    2017-02-01

    Liver transplant recipients are at risk of invasive fungal infections, especially candidiasis. Echinocandin is recommended as prophylactic treatment but is increasingly associated with resistance. Our aim was to assess echinocandin drug resistance in Candida spp. isolated from liver transplant recipients treated with this antifungal class. For this, all liver-transplanted patients in a University Hospital (Créteil, France) between January and June of 2013 and 2015 were included. Susceptibilities of Candida isolates to echinocandins were tested by Etest and the EUCAST reference method. Isolates were analyzed by FKS sequencing and genotyped based on microsatellites or multilocus sequence typing (MLST) profiles. Ninety-four patients were included, and 39 patients were colonized or infected and treated with echinocandin. Echinocandin resistance appeared in 3 (8%) of the treated patients within 1 month of treatment. One patient was colonized by resistant Candida glabrata, one by resistant Candida dubliniensis, and one by resistant Candida albicans Molecular analysis found three mutations in FKS2 HS1 (F659S, S663A, and D666E) for C. glabrata and one mutation in FKS1 HS1 (S645P) for C. dubliniensis and C. albicans Susceptible and resistant isolates belonged to the same genotype. To our knowledge, this is the first study on echinocandin resistance in Candida spp. in a liver transplant population. Most resistant isolates were found around/in digestive sites, perhaps due to lower diffusion of echinocandin in these sites. This work documents the risk of emergence of resistance to echinocandin, even after short-term treatment. Copyright © 2017 Prigent et al.

  18. Functionality of whey protein isolates and hydrolyzed whey proteins

    OpenAIRE

    Zoran Herceg; Vesna Lelas; Greta Krešić; Anet Režek

    2005-01-01

    The aim of this study was to determine functional properties: solubility, dispersibility, viscosity, emulsifiying, foaming and rheological properties of hydrolyzed whey protein (HWP) and whey protein isolate (WPI). Particle size analysis and specific area of HWP and WPI were performed by «Mie – theory» of «light scatering» using «Malvern Mastersizer X». The results of this analysis have shown that HWP had higher particle size and specific area than WPI.By examing functional properties, it has...

  19. Passive earthquake-resistance through base isolation

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Ting-Shu; Seidensticker, R.W.

    1990-04-01

    Base isolation is an effective approach in mitigating the seismic forces transmitted to the superstructure. It is able to provide reliable protection for the superstructure, its contents and occupants. With base isolation, dynamic characteristics of the superstructure and its contents become more predictable and controllable. In recent years, different base isolation systems have been installed in various new and existing buildings, including office buildings, computer centers, buildings for high technology industries, emergency and communication centers, buildings for high technology industries, emergency and communication centers. The concept of seismic isolation is also under active investigation for nuclear power plants, where safety and reliability of these systems are of utmost importance. One such system uses laminated high-damping rubber and steel shim plates vulcanized into a solid bearing. Under a joint program between Shimizu of Japan and Argonne National Laboratory of the United States, such high-damping rubber bearings were installed in the spring of 1989 in a three-story full size test building at the Tohoku University, Sendai, Japan. Within a period of six months after their installation, this test building and the bearings have experienced more than fifteen earthquakes. Valuable information on the performance of the isolation bearings and the building has been (and is continuing to be) gathered and analyzed. Simulation studies of the effects of these earthquakes using lumped mass systems have been conducted, and are in close agreement with the actual observed responses. 3 refs., 11 figs.

  20. Antibiotic resistance of lactic acid bacteria isolated from Chinese yogurts.

    Science.gov (United States)

    Zhou, N; Zhang, J X; Fan, M T; Wang, J; Guo, G; Wei, X Y

    2012-09-01

    The aim of this study was to evaluate the susceptibility of 43 strains of lactic acid bacteria, isolated from Chinese yogurts made in different geographical areas, to 11 antibiotics (ampicillin, penicillin G, roxithromycin, chloramphenicol, tetracycline, chlortetracycline, lincomycin, kanamycin, streptomycin, neomycin, and gentamycin). The 43 isolates (18 Lactobacillus bulgaricus and 25 Streptococcus thermophilus) were identified at species level and were typed by random amplified polymorphic DNA analysis. Thirty-five genotypically different strains were detected and their antimicrobial resistance to 11 antibiotics was determined using the agar dilution method. Widespread resistance to ampicillin, chloramphenicol, chlortetracycline, tetracyclines, lincomycin, streptomycin, neomycin, and gentamycin was found among the 35 strains tested. All of the Strep. thermophilus strains tested were susceptible to penicillin G and roxithromycin, whereas 23.5 and 64.7% of Lb. bulgaricus strains, respectively, were resistant. All of the Strep. thermophilus and Lb. bulgaricus strains were found to be resistant to kanamycin. The presence of the corresponding resistance genes in the resistant isolates was investigated through PCR, with the following genes detected: tet(M) in 1 Lb. bulgaricus and 2 Strep. thermophilus isolates, ant(6) in 2 Lb. bulgaricus and 2 Strep. thermophilus isolates, and aph(3')-IIIa in 5 Lb. bulgaricus and 2 Strep. thermophilus isolates. The main threat associated with these bacteria is that they may transfer resistance genes to pathogenic bacteria, which has been a major cause of concern to human and animal health. To our knowledge, the aph(3')-IIIa and ant(6) genes were found in Lb. bulgaricus and Strep. thermophilus for the first time. Further investigations are required to analyze whether the genes identified in Lb. bulgaricus and Strep. thermophilus isolates might be horizontally transferred to other species.

  1. [Antibiotic resistance and virulence factors in clinical Salmonella enterica isolates].

    Science.gov (United States)

    de Toro, María; Seral, Cristina; Rojo-Bezares, Beatriz; Torres, Carmen; Castillo, F Javier; Sáenz, Yolanda

    2014-01-01

    The increase of Salmonella enterica isolates multi-resistant to different antibiotics, including β-lactams and fluoroquinolones, is a problem of clinical importance. The dissemination of Salmonella Typhimurium resistant to ampicillin (AMP)-chloramphenicol (CHL)-streptomycin (STR)-sulphonamides and (SUL)-tetracycline (TET), that harbour the Salmonella Genomic Island type 1 (SGI1), and the acquisition of transferable genetic material have favoured the multi-resistance in this genus. A total of 114 clinical S.enterica isolates were studied (period 2009-2010). The susceptibility to 20 antibiotics was determined by disc diffusion and microdilution. The antimicrobial resistance mechanisms and the integrons were analysed by PCR, and sequencing in the AMP(R) isolates. In all the blaPSE-1-positive isolates, the clonal relationship was determined by PFGE, as well as the presence of SGI1 and 29 virulence genes by PCR. Eighteen different serotypes were found among the 114 isolates studied, Typhimurium (61%) and Enteritidis (16%) being the most prevalent. High percentages of resistance to SUL (68%), TET (58%), AMP (55%) and STR (46%) were observed. The great majority (92%) of 63 AMP(R) isolates were multi-resistant, with the AMP-STR-TET-SUL phenotype (19 isolates) being the most frequent one and associated with the blaTEM-1b+strA-strB+tet(B)+sul2 genotype. Class 1 integrons (7 different structures) were observed in 48% AMP(R) isolates, highlighting the blaOXA-1+aadA1 structure (8 isolates), one empty integron and non-classical integrons (5 isolates). The blaPSE-1 gene was detected inside the classical SGI1 structure in 13 clonally-related isolates that showed the same virulence profile. The high percentage of multi-resistant S.enterica isolates, especially associated to S.Typhimurium, to the AMP, STR, TET and SUL phenotype, and to the blaTEM-1b+strA-strB+tet(B)+sul2 genotype, shows an important risk of possible failures in the treatment of serious infections caused by this

  2. Transporter protein and drug resistance of Trypanosoma.

    Science.gov (United States)

    Medina, Noraine P; Mingala, Claro N

    2016-01-01

    Trypanosoma infection is one of the most important infections in livestock and humans. One of the main problems of its therapeutic control and treatment is the resurgence of drug resistance. One of the most studied causes of such resistance is the function of its adenosine transporter gene. A trypanosomal gene TbAT1 from Trypanosoma brucei has been cloned in yeast to demonstrate its function in the transport of adenosine and trypanocidal agents. Drug resistant trypanosomes showed a defective TbAT1 variant; furthermore, deletion of the gene and set point mutations in the transporter gene has been demonstrated from isolates from relapse patients. The molecular understanding of the mechanism of action trypanocidal agents and function of transporter gene can lead to control of drug resistance of Trypanosomes.

  3. Composition and functional properties of Lupinus campestris protein isolates.

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    Rodríguez-Ambriz, S L; Martínez-Ayala, A L; Millán, F; Dávila-Ortíz, G

    2005-09-01

    Protein isolates from L. campestris and soybean seeds were prepared using isoelectric precipitation (PI) and micellization (MI) procedures. The amount of protein recovered was considerably higher with the isoelectric precipitation than with the micellization procedure (60% and 30%, respectively). Protein contents were higher than 90% in protein isolates. Antinutritional factors content (alkaloids, lectins, and tannins) were reduced to innocuous levels after protein isolate preparation. Minimum protein solubility for the precipitated lupin protein isolate (LPI) was at pH 4.0, and between pH 4 and 6 for the micellized lupin protein isolate (LMI), increasing at both extremes of the pH scale. Water absorption for the LMI was 1.3 ml/g of protein and its oil absorption 2.2 ml/g of protein. The LPI had 1.7 ml/g of protein in both water and oil absorption. Foaming capacity and stability was pH-dependent. Foaming capacity was higher at pH 2 and lower near the protein isoelectric points. Minimum protein concentration for gelation in LMI was 8% w/v at pH 4, while for LPI was 6% at pH 4 and 6. Amino acid composition in L. campestris flour and protein isolates was high in lysine and low in methionine. Most of the essential amino acids in lupin protein isolates were at acceptable levels compared to a reference pattern for infants and adults. The electrophoretic pattern of both protein isolates showed three bands with different mobilities, suggesting that the protein fractions belong to alpha-conglutin (11S-like protein), beta-conglutin (7S-like protein) and gamma-conglutin. It is proven that some of the functional properties of L. campestris protein isolates are similar to those soybean protein isolates recovered under equal conditions.

  4. Biofilm Production in Carbapenem Resistant Isolates from Chronic Wound Infections

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    Swarna SR

    2017-02-01

    Full Text Available Biofilms are communities of microorganisms covered with extracellular polymeric substances. Such biofilm phenotype makes the microorganism resistant to antibiotics and plays a role in wound chronicity. This results in prolonged hospital stays in ICU, greater cost, and increased mortality. Methods: Pus swabs (59 were collected from a tertiary care hospital near Chennai were processed and identified using standard procedure followed by antibiotic susceptibility testing and identification of carbapenem resistance by Modified Hodge test as per CLSI guidelines. The biofilm formation was tested using plastic microtiter plate method. Results: Out of 59 pus swabs, 51 yielded growth with 69 isolates and 8 yielded no growth. Among the 69 isolates, 51 were GNB and 18 were GPC. Biofilm detection was noted in 84.31% (43/51 GNB isolates with 0.1% crystal violet whereas 100% (51/51 showed biofilm positive with 0.1% safranin. About 74.50% (38/51 isolates of GNB were carbapenem resistant by screening with disk diffusion method. Only 24% (6/25 of GNB isolates among Enterobacteriaceae were positive by Modified Hodge test method. Conclusion: The result shows the association of biofilm production among carbapenem resistant isolates obtained from chronic wound infections.

  5. Molecular resistance mechanisms of macrolide-resistant invasive Streptococcus pneumoniae isolates from Alaska, 1986 to 2010.

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    Rudolph, Karen; Bulkow, Lisa; Bruce, Michael; Zulz, Tammy; Reasonover, Alisa; Harker-Jones, Marcella; Hurlburt, Debby; Hennessy, Thomas

    2013-11-01

    The rapid emergence of antibiotic-resistant pneumococcal strains has reduced treatment options. The aim of this study was to determine antimicrobial susceptibilities, serotype distributions, and molecular resistance mechanisms among macrolide-resistant invasive pneumococcal isolates in Alaska from 1986 to 2010. We identified cases of invasive pneumococcal disease in Alaska from 1986 to 2010 through statewide population-based laboratory surveillance. All invasive pneumococcal isolates submitted to the Arctic Investigations Program laboratory were confirmed by standard microbiological methods and serotyped by slide agglutination and the Quellung reaction. MICs were determined by the broth microdilution method, and macrolide-resistant genotypes were determined by multiplex PCR. Among 2,923 invasive pneumococcal isolates recovered from 1986 to 2010, 270 (9.2%) were nonsusceptible to erythromycin; 177 (66%) erythromycin-nonsusceptible isolates demonstrated coresistance to penicillin, and 167 (62%) were multidrug resistant. The most frequent serotypes among the macrolide-resistant isolates were serotypes 6B (23.3%), 14 (20.7%), 19A (16.7%), 9V (8.9%), 19F (6.3%), 6A (5.6%), and 23F (4.8%). mef and erm(B) genes were detected in 207 (77%) and 32 (12%) of the isolates, respectively. Nineteen (7%) of the erythromycin-nonsusceptible isolates contained both mef and erm(B) genotypes; 15 were of serotype 19A. There was significant year-to-year variation in the proportion of isolates that were nonsusceptible to erythromycin (P resistance among pneumococcal isolates from Alaska is mediated predominantly by mef genes, and this has not changed significantly over time. However, there was a statistically significant increase in the proportion of isolates that possess both erm(B) and mef, primarily due to serotype 19A isolates.

  6. Antimicrobial resistance of Actinobacillus pleuropneumoniae isolated from swine.

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    Vanni, Michele; Merenda, Marianna; Barigazzi, Giuseppe; Garbarino, Chiara; Luppi, Andrea; Tognetti, Rosalba; Intorre, Luigi

    2012-04-23

    The aim of this retrospective study was to evaluate the antimicrobial resistance rates and the trend in resistance of Actinobacillus pleuropneumoniae isolated from pigs in Italy from 1994 to 2009. A total of 992 A. pleuropneumoniae isolates were tested for their susceptibility to a panel of antimicrobial agents in a disk diffusion method. Resistance to 7 drugs (amoxicillin, amoxicillin/clavulanic acid, ampicillin, cefquinome, cotrimoxazole, penicillin G and tilmicosin) showed a significant increasing trend over the time, while for 2 drugs (gentamycin and marbofloxacin) a significant decrease was observed. Resistance to the remaining 14 antimicrobial agents tested did not change significantly over the study period. Most of the isolates retained high susceptibility to antimicrobials usually effective against A. pleuropneumoniae such as amphenicols, fluoroquinolones and ceftiofur. However, high rates of resistance were observed for potentiated sulfa drugs, tetracyclines and penicillins which are currently recommended antimicrobials for pig pleuropneumonia therapy. Our results suggest the importance of continued monitoring of A. pleuropneumoniae clinical isolates in order to choose the most appropriate treatment of infections and to control the increase of resistance to currently used antimicrobials. Copyright © 2011 Elsevier B.V. All rights reserved.

  7. Antibiotic resistance and virulence genes in coliform water isolates.

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    Stange, C; Sidhu, J P S; Tiehm, A; Toze, S

    2016-11-01

    Widespread fecal pollution of surface water may present a major health risk and a significant pathway for dissemination of antibiotic resistance bacteria. The River Rhine is one of the longest and most important rivers in Europe and an important raw water source for drinking water production. A total of 100 coliform isolates obtained from River Rhine (Germany) were examined for their susceptibility to seven antimicrobial agents. Resistances against amoxicillin, trimethoprim/sulfamethoxazole and tetracycline were detected in 48%, 11% and 9% of isolates respectively. The antibiotic resistance could be traced back to the resistance genes blaTEM, blaSHV, ampC, sul1, sul2, dfrA1, tet(A) and tet(B). Whereby, the ampC gene represents a special case, because its presence is not inevitably linked to a phenotypic antibiotic resistance. Multiple antibiotics resistance was often accompanied by the occurrence of class 1 or 2 integrons. E. coli isolates belonging to phylogenetic groups A and B1 (commensal) were more predominant (57%) compared to B2 and D groups (43%) which are known to carry virulent genes. Additionally, six E. coli virulence genes were also detected. However, the prevalence of virulence genes in the E. coli isolates was low (not exceeding 4.3% per gene) and no diarrheagenic E. coli pathotypes were detected. This study demonstrates that surface water is an important reservoir of ARGs for a number of antibiotic classes such as sulfonamide, trimethoprim, beta-lactam-antibiotics and tetracycline. The occurrence of antibiotic resistance in coliform bacteria isolated from River Rhine provides evidence for the need to develop management strategies to limit the spread of antibiotic resistant bacteria in aquatic environment. Copyright © 2016 Elsevier GmbH. All rights reserved.

  8. Isolation and resistance test of several isolates of Lactobacillus in low pH

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    RIANI HARDININGSIH

    2006-01-01

    Full Text Available Several isolates of Lactobacillus had been isolated from Indonesian fermented foods. Four isolates where isolated from tongcai (TT2, saguer drink (Sg.Mnd.N2, pindang ikan selar (PSl1 and sawi asin (S5 had been used in this research. The aim of the research was to get Lactobacillus isolates as probiotic candidate and to know its resistance to low pH. The treatment were several concentration of low pH of MRS broth medium, i.e. 2; 2.5; 3; and 6.5 (optimal pH. The optical density (OD was measured after 24, 48, 72, and 96 hours respectively, with three replication by using spectrophotometer ( = 600 nm. The results showed that all Lactobacillus isolates (i.e. TT2, Sg.Mnd.N2, PSl1, and S5 could been used as probiotic candidate because they were resistant to all of low pH used.

  9. Fluoroquinolone resistance mechanisms of Shigella flexneri isolated in Bangladesh.

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    Ishrat J Azmi

    Full Text Available To investigate the prevalence and mechanisms of fluoroquinolone resistance in Shigella species isolated in Bangladesh and to compare with similar strains isolated in China.A total of 3789 Shigella isolates collected from Clinical Microbiology Laboratory of icddr,b, during 2004-2010 were analyzed for antibiotic susceptibility. Analysis of plasmids, plasmid-mediated quinolone-resistance genes, PFGE, and sequencing of genes of the quinolone-resistance-determining regions (QRDR were conducted in representative strains isolated in Bangladesh and compared with strains isolated in Zhengding, China. In addition, the role of efflux-pump was studied by using the efflux-pump inhibitor carbonyl cyanide-m-chlorophenylhydrazone (CCCP.Resistance to ciprofloxacin in Shigella species increased from 0% in 2004 to 44% in 2010 and S. flexneri was the predominant species. Of Shigella spp, ciprofloxacin resistant (CipR strains were mostly found among S. flexneri (8.3%, followed by S. sonnei (1.5%. Within S. flexneri (n = 2181, 14.5% were resistance to ciprofloxacin of which serotype 2a was predominant (96%. MIC of ciprofloxacin, norfloxacin, and ofloxacin were 6-32 mg/L, 8-32 mg/L, and 8-24 mg/L, respectively in S. flexneri 2a isolates. Sequencing of QRDR genes of resistant isolates showed double mutations in gyrA gene (Ser83Leu, Asp87Asn/Gly and single mutation in parC gene (Ser80Ile. A difference in amino acid substitution at position 87 was found between strains isolated in Bangladesh (Asp87Asn and China (Asp87Gly except for one. A novel mutation at position 211 (His→Tyr in gyrA gene was detected only in the Bangladeshi strains. Susceptibility to ciprofloxacin was increased by the presence of CCCP indicating the involvement of energy dependent active efflux pumps. A single PFGE type was found in isolates from Bangladesh and China suggesting their genetic relatedness.Emergence of fluoroquinolone resistance in Shigella undermines a major challenge in current

  10. Fluoroquinolone Resistance Mechanisms of Shigella flexneri Isolated in Bangladesh

    Science.gov (United States)

    Azmi, Ishrat J.; Khajanchi, Bijay K.; Akter, Fatema; Hasan, Trisheeta N.; Shahnaij, Mohammad; Akter, Mahmuda; Banik, Atanu; Sultana, Halima; Hossain, Mohammad A.; Ahmed, Mohammad K.; Faruque, Shah M.; Talukder, Kaisar A.

    2014-01-01

    Objective To investigate the prevalence and mechanisms of fluoroquinolone resistance in Shigella species isolated in Bangladesh and to compare with similar strains isolated in China. Methods A total of 3789 Shigella isolates collected from Clinical Microbiology Laboratory of icddr,b, during 2004–2010 were analyzed for antibiotic susceptibility. Analysis of plasmids, plasmid-mediated quinolone-resistance genes, PFGE, and sequencing of genes of the quinolone-resistance-determining regions (QRDR) were conducted in representative strains isolated in Bangladesh and compared with strains isolated in Zhengding, China. In addition, the role of efflux-pump was studied by using the efflux-pump inhibitor carbonyl cyanide-m-chlorophenylhydrazone (CCCP). Results Resistance to ciprofloxacin in Shigella species increased from 0% in 2004 to 44% in 2010 and S. flexneri was the predominant species. Of Shigella spp, ciprofloxacin resistant (CipR) strains were mostly found among S. flexneri (8.3%), followed by S. sonnei (1.5%). Within S. flexneri (n = 2181), 14.5% were resistance to ciprofloxacin of which serotype 2a was predominant (96%). MIC of ciprofloxacin, norfloxacin, and ofloxacin were 6–32 mg/L, 8–32 mg/L, and 8–24 mg/L, respectively in S. flexneri 2a isolates. Sequencing of QRDR genes of resistant isolates showed double mutations in gyrA gene (Ser83Leu, Asp87Asn/Gly) and single mutation in parC gene (Ser80Ile). A difference in amino acid substitution at position 87 was found between strains isolated in Bangladesh (Asp87Asn) and China (Asp87Gly) except for one. A novel mutation at position 211 (His→Tyr) in gyrA gene was detected only in the Bangladeshi strains. Susceptibility to ciprofloxacin was increased by the presence of CCCP indicating the involvement of energy dependent active efflux pumps. A single PFGE type was found in isolates from Bangladesh and China suggesting their genetic relatedness. Conclusions Emergence of fluoroquinolone resistance in Shigella

  11. Antibiotic resistance is prevalent in an isolated cave microbiome.

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    Kirandeep Bhullar

    Full Text Available Antibiotic resistance is a global challenge that impacts all pharmaceutically used antibiotics. The origin of the genes associated with this resistance is of significant importance to our understanding of the evolution and dissemination of antibiotic resistance in pathogens. A growing body of evidence implicates environmental organisms as reservoirs of these resistance genes; however, the role of anthropogenic use of antibiotics in the emergence of these genes is controversial. We report a screen of a sample of the culturable microbiome of Lechuguilla Cave, New Mexico, in a region of the cave that has been isolated for over 4 million years. We report that, like surface microbes, these bacteria were highly resistant to antibiotics; some strains were resistant to 14 different commercially available antibiotics. Resistance was detected to a wide range of structurally different antibiotics including daptomycin, an antibiotic of last resort in the treatment of drug resistant Gram-positive pathogens. Enzyme-mediated mechanisms of resistance were also discovered for natural and semi-synthetic macrolide antibiotics via glycosylation and through a kinase-mediated phosphorylation mechanism. Sequencing of the genome of one of the resistant bacteria identified a macrolide kinase encoding gene and characterization of its product revealed it to be related to a known family of kinases circulating in modern drug resistant pathogens. The implications of this study are significant to our understanding of the prevalence of resistance, even in microbiomes isolated from human use of antibiotics. This supports a growing understanding that antibiotic resistance is natural, ancient, and hard wired in the microbial pangenome.

  12. Isolation and characterization of bacteria resistant to metallic copper surfaces.

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    Santo, Christophe Espírito; Morais, Paula Vasconcelos; Grass, Gregor

    2010-03-01

    Metallic copper alloys have recently attracted attention as a new antimicrobial weapon for areas where surface hygiene is paramount. Currently it is not understood on a molecular level how metallic copper kills microbes, but previous studies have demonstrated that a wide variety of bacteria, including Escherichia coli, Staphylococcus aureus, and Clostridium difficile, are inactivated within minutes or a few hours of exposure. In this study, we show that bacteria isolated from copper alloy coins comprise strains that are especially resistant against the toxic properties exerted by dry metallic copper surfaces. The most resistant of 294 isolates were Gram-positive staphylococci and micrococci, Kocuria palustris, and Brachybacterium conglomeratum but also included the proteobacterial species Sphingomonas panni and Pseudomonas oleovorans. Cells of some of these bacterial strains survived on copper surfaces for 48 h or more. Remarkably, when these dry-surface-resistant strains were exposed to moist copper surfaces, resistance levels were close to those of control strains and MICs for copper ions were at or below control strain levels. This suggests that mechanisms conferring resistance against dry metallic copper surfaces in these newly isolated bacterial strains are different from well-characterized copper ion detoxification systems. Furthermore, staphylococci on coins did not exhibit increased levels of resistance to antibiotics, arguing against coselection with copper surface resistance traits.

  13. Isolation and characterization of cidofovir resistant vaccinia viruses

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    Prichard Mark N

    2008-05-01

    Full Text Available Abstract Background The emergence of drug resistant viruses, together with the possibility of increased virulence, is an important concern in the development of new antiviral compounds. Cidofovir (CDV is a phosphonate nucleotide that is approved for use against cytomegalovirus retinitis and for the emergency treatment of smallpox or complications following vaccination. One mode of action for CDV has been demonstrated to be the inhibition of the viral DNA polymerase. Results We have isolated several CDV resistant (CDVR vaccinia viruses through a one step process, two of which have unique single mutations within the DNA polymerase. An additional resistant virus isolate provides evidence of a second site mutation within the genome involved in CDV resistance. The CDVR viruses were 3–7 fold more resistant to the drug than the parental viruses. The virulence of the CDVR viruses was tested in mice inoculated intranasally and all were found to be attenuated. Conclusion Resistance to CDV in vaccinia virus can be conferred individually by at least two different mutations within the DNA polymerase gene. Additional genes may be involved. This one step approach for isolating resistant viruses without serial passage and in the presence of low doses of drug minimizes unintended secondary mutations and is applicable to other potential antiviral agents.

  14. PHENOTYPIC AND MOLECULAR CHARACTERIZATION OF ANTIBIOTICS RESISTANCE E. CLOACAE ISOLATES

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    Abdalnabi J Abid

    2015-12-01

    Full Text Available The present study aimed detecting and characterizing of β-lactamases producing E.cloacae isolated from different clinical sources in Hilla hospitals using phenotypic and molecular methods. A total of 308 samples were collected from two major hospitals at Hilla Province from October 2013 to April 2014. All isolates were tested biochemically, it was found that only 15 isolates from all isolates were belonging to Enterobactercloacae. All E. cloacae isolates were primarily screened for β-lactams resistance. Antibiotic susceptibility and minimum inhibitory concentration tests were performed using disk diffusion and agar dilution methods, respectively. The molecular study documented a widespread of Amp C genes among isolates of E. cloacae isolatesrepresented by 6/15(40% positive isolates for Amp C primers. PCR assay revealed that prevalence rate of bla-TEM gene among tested isolates was 9(60%. followed by the bla-OXA gene was detected only in 3(20%.While bla-VEB gene and bla-SHV gene was not detected in any of the isolates. Some virulence factors of bacteria were also studied, and the results showed that all bacterial strains have capsule ,the results also also detected biofilm formation among isolates and the results revealed that 13(86%of the isolates are biofilm former.

  15. Characterization of mannitol-fermenting methicillin-resistant staphylococci isolated from pigs in Nigeria

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    Clifford C. Ugwu

    2015-09-01

    Full Text Available This study was conducted to determine the species distribution, antimicrobial resistance pheno- and genotypes and virulence traits of mannitol-positive methicillin-resistant staphylococci (MRS isolated from pigs in Nsukka agricultural zone, Nigeria. Twenty mannitol-positive methicillin-resistant coagulase-negative staphylococcal (MRCoNS strains harboring the mecA gene were detected among the 64 Staphylococcus isolates from 291 pigs. A total of 4 species were identified among the MRCoNS isolates, namely, Staphylococcus sciuri (10 strains, Staphylococcus lentus (6 strains, Staphylococcus cohnii (3 strains and Staphylococcus haemolyticus (one strain. All MRCoNS isolates were multidrug-resistant. In addition to β-lactams, the strains were resistant to fusidic acid (85%, tetracycline (75%, streptomycin (65%, ciprofloxacin (65%, and trimethoprim/sulphamethoxazole (60%. In addition to the mecA and blaZ genes, other antimicrobial resistance genes detected were tet(K, tet(M, tet(L, erm(B, erm(C, aacA-aphD, aphA3, str, dfrK, dfrG, catpC221, and catpC223. Thirteen isolates were found to be ciprofloxacin-resistant, and all harbored a Ser84Leu mutation within the QRDR of the GyrA protein, with 3 isolates showing 2 extra substitutions, Ser98Ile and Arg100Lys (one strain and Glu88Asp and Asp96Thr (2 strains. A phylogenetic tree of the QRDR nucleotide sequences in the gyrA gene revealed a high nucleotide diversity, with several major clusters not associated with the bacterial species. Our study highlights the possibility of transfer of mecA and other antimicrobial resistance genes from MRCoNS to pathogenic bacteria, which is a serious public health and veterinary concern.

  16. Antimicrobial resistance and virulence markers in methicillin sensitive Staphylococcus aureus isolates associated with nasal colonization.

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    Sarkar, Abdullah; Raji, Adeola; Garaween, Ghada; Soge, Olusegun; Rey-Ladino, Jose; Al-Kattan, Wael; Shibl, Atef; Senok, Abiola

    2016-04-01

    Most Staphylococcus aureus infections occur in previously colonized persons who also act as reservoirs for continued dissemination. This study aimed to investigate the carriage of antimicrobial resistance and virulence markers in S. aureus isolates associated with nasal colonization. The study was conducted from December 2013-April 2014. Nasal swabs were collected and questionnaires administered to 97 medical students in Riyadh Saudi Arabia. Bacterial culture, identification and antimicrobial susceptibility testing were performed by conventional methods and chromogenic agar was used for methicillin resistant S. aureus (MRSA) screening. Molecular characterization of isolates was carried out using the StaphyType DNA microarray. Thirty two students (43%) had S. aureus nasal carriage (MSSA = 31; MRSA = 1). Seventeen clonal complexes (CC) were identified namely: CC15-MSSA (n = 5), CC1-MSSA-SCCfus (n = 4), CC8-MSSA (n = 3), CC22-MSSA (n = 3), CC25-MSSA (n = 3), CC101-MSSA (n = 2). Other CC found as single isolates were CC5-MSSA, CC6-MSSA, CC30-MSSA, CC45-MSSA, CC96-MSSA, CC188-MSSA, CC398-MSSA, CC942-MSSA/PVL+, CC1290-MSSA, ST2482-MSSA, CC80-MRSA-IV/PVL+. The CC1-SCCfus isolates harbored the Staphylococcal cassette chromosome (SCC) with ccrA-1; ccrB-1 and ccrB-3 genes plus the putative fusidic acid resistance marker Q6GD50. One MSSA isolate was genotyped as coagulase negative Staphylococcus spp with an irregular composite SCCmec element. Majority of the isolates harbored various virulence genes including the hemolysin, enterotoxin, and exfoliative genes as well as various adhesive protein producing genes. Although there was low carriage of MRSA, the MSSA isolates harbored various resistance and virulence genes including those usually seen in MRSA isolates. The presence of isolates with incomplete SCCmec elements plus putative resistance and virulence genes is of concern.

  17. Genotypes and antibiotic resistance of canine Campylobacter jejuni isolates.

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    Amar, Chantal; Kittl, Sonja; Spreng, David; Thomann, Andreas; Korczak, Bożena M; Burnens, André P; Kuhnert, Peter

    2014-01-10

    Campylobacter jejuni is the most important cause of bacterial gastroenteritis in humans. It is a commensal in many wild and domestic animals, including dogs. Whereas genotypes of human and chicken C. jejuni isolates have been described in some detail, only little information on canine C. jejuni genotypes is available. To gain more information on genotypes of canine C. jejuni and their zoonotic potential, isolates from routine diagnostics of diarrheic dogs as well as isolates of a prevalence study in non-diarrheic dogs were analyzed. Prevalence of thermophilic Campylobacter among non-diarrheic dogs was 6.3% for C. jejuni, 5.9% for Campylobacter upsaliensis and 0.7% for Campylobacter coli. The C. jejuni isolates were genotyped by multi locus sequence typing (MLST) and flaB typing. Resistance to macrolides and quinolones was genetically determined in parallel. Within the 134 genotyped C. jejuni isolates 57 different sequence types (ST) were found. Five STs were previously unrecognized. The most common STs were ST-48 (11.2%), ST-45 (10.5%) and ST-21 (6.0%). Whereas no macrolide resistance was found, 28 isolates (20.9%) were resistant to quinolones. ST-45 was significantly more prevalent in diarrheic than in non-diarrheic dogs. Within the common time frame of isolation 94% of the canine isolates had a ST that was also found in human clinical isolates. In conclusion, prevalence of C. jejuni in Swiss dogs is low but there is a large genetic overlap between dog and human isolates. Given the close contact between human and dogs, the latter should not be ignored as a potential source of human campylobacteriosis. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Multidrug resistant bacteria isolated from septic arthritis in horses

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    Rodrigo G. Motta

    Full Text Available ABSTRACT: Septic arthritis is a debilitating joint infectious disease of equines that requires early diagnosis and immediate therapeutic intervention to prevent degenerative effects on the articular cartilage, as well as loss of athletic ability and work performance of the animals. Few studies have investigated the etiological complexity of this disease, as well as multidrug resistance of isolates. In this study, 60 horses with arthritis had synovial fluid samples aseptically collected, and tested by microbiological culture and in vitro susceptibility test (disk diffusion using nine antimicrobials belonging to six different pharmacological groups. Bacteria were isolated in 45 (75.0% samples, as follows: Streptococcus equi subsp. equi (11=18.3%, Escherichia coli (9=15.0%, Staphylococcus aureus (6=10.0%, Streptococcus equi subsp. zooepidemicus (5=8.3%, Staphylococcus intermedius (2=3.3%, Proteus vulgaris (2=3.3%, Trueperella pyogenes (2=3.3%, Pseudomonas aeruginosa (2=3.3%, Klebsiella pneumoniae (1=1.7%, Rhodococcus equi (1=1.7%, Staphylococcus epidermidis (1=1.7%, Klebsiella oxytoca (1=1.7%, Nocardia asteroides (1=1.7%, and Enterobacter cloacae (1=1.7%. Ceftiofur was the most effective drug (>70% efficacy against the pathogens in the disk diffusion test. In contrast, high resistance rate (>70% resistance was observed to penicillin (42.2%, enrofloxacin (33.3%, and amikacin (31.2%. Eleven (24.4% isolates were resistant to three or more different pharmacological groups and were considered multidrug resistant strains. The present study emphasizes the etiological complexity of equine septic arthritis, and highlights the need to institute treatment based on the in vitro susceptibility pattern, due to the multidrug resistance of isolates. According to the available literature, this is the first report in Brazil on the investigation of the etiology. of the septic arthritis in a great number of horses associated with multidrug resistance of the isolates.

  19. CHARACTERISATION OF METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS ISOLATES FROM SHINGLES PATIENTS

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    Jasmine R. et al.

    2011-11-01

    Full Text Available Even after treating Shingles patients with antiviral drugs, they are found to suffer from secondary bacterial infections. With this background as a guide, we undertook an investigation to isolate the bacterial pathogens from the pus of Shingles patients. Among the isolates obtained during the one year study period, Staphylococcus aureus sp. was found to be multi drug resistant and hence it was chosen for the study. The antibiogram pattern of the methicillin resistant S. aureus was obtained, since this could serve as a tool for suggesting useful drugs.

  20. Selection and characterisation of monepantel resistance in Teladorsagia circumcincta isolates

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    D.J. Bartley

    2015-08-01

    To address these questions three MPTL-resistant Teladorsagia circumcincta isolates (MTci2-11, MTci5-13 and MTci7-12 have been artificially selected in vivo from phenotypically characterised parent isolates (MTci2, MTci5, MTci7 respectively. The selection process involved collecting and culturing eggs from surviving worms from sheep administered sub-optimal dosages of MPTL (Zolvix® to provide infective larvae to infect further sheep until resistant isolates were generated (between 9 and 13 rounds of selection. A controlled efficacy test was conducted using the original parental isolates and the newly generated MPTL resistant isolates (n = 5 per group. Selected isolates were assessed both under anthelmintic stress (Zolvix®, 2.5 mg/kg bodyweight; MTci-MPTL and at rest (untreated, MTci-CON. A number of life-history traits were assessed, namely, worm establishment rates, time to patency, faecal egg output, body length of adults and eggs in utero. The estimated resistance status of the selected isolates was confirmed with 48%, 28% and 9% reductions in worm burden at 7-days post Zolvix® administration for MTci2-11-MPTL, MTci5-13-MPTL and MTci7-12-MPTL, respectively, compared with untreated controls. One of the selected isolates MTci7-12-CON showed significantly greater total worm burden (p = 0.025, greater establishment rate (p = 0.033, decreased time to patency (p = 0.048, higher cumulative egg outputs (p = 0.002 compared with its parental derivative MTci7. The trial results suggest that anthelmintic selection in T. circumcincta, albeit under experimental conditions, can select for more prolific/fecund and quicker maturing populations. These data provide an insight into how parasites evolve in response to anthelmintic pressure.

  1. Plasmid mediated antibiotic resistance in isolated bacteria from burned patients.

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    Beige, Fahimeh; Baseri Salehi, Majid; Bahador, Nima; Mobasherzadeh, Sina

    2015-01-01

    Nowadays, the treatment of burned patients is difficult because of the high frequency of infection with antibiotic resistance bacteria. This study was conducted to evaluate the level of antibiotic resistance in Gram-negative bacteria and its relation with the existence of plasmid. The samples were collected from two hundred twenty hospitalized burned patients in Isfahan burn hospital during a three-month period (March 2012 to June 2012). The samples were isolated and the Gram-negative bacteria were identified using phenotypic method and API 20E System. Antibiotic susceptibility and plasmid profile were determined by standard Agar disc diffusion and plasmid spin column extraction methods. Totally 117 Gram-negative bacteria were isolated, the most common were Pseudomonas aerugionsa (37.6%), P. fluorescens (25.6%), Acinetobacter baumanii (20/5%) and Klebsiella pneumoniae (7.6%), respectively. The isolates showed high frequency of antibiotic resistance against ceftazidime and co-amoxiclave (100%) and low frequency of antibiotic resistance against amikacin with (70%).The results indicated that 60% of the isolates harboured plasmid. On the other hand, the patients infected with A. baumanii and P. aeruginosa were cured (with 60% frequency) whereas, those infected with P. fluorescens were not cured. Hence, probably antibiotic resistance markers of A. baumanii and P. aeruginosa are plasmid mediated; however, P. fluorescens is chromosomally mediated. Based on our findings, P. aerugionsa is a major causative agent of wound infections and amikacin could be considered as a more effective antibiotic for treatment of the burned patients.

  2. Antimicrobial Resistance of Staphylococcal Strains Isolated from Various Pathological Products

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    Laura-Mihaela SIMON

    2010-12-01

    Full Text Available Background: The optimal choice of antimicrobial therapy is an important problem in hospital environment in which the selection of resistant and virulent strains easy occurs. S. aureus and especially MRSA(methicillin-resistant S. aureus creates difficulties in both treatment and prevention of nosocomial infections. Aim: The purpose of this study is to determine the sensitivity and the resistance to chemotherapy of staphylococci strains isolated from various pathological products. Material and Method: We identified Staphylococccus species after morphological appearance, culture properties, the production of coagulase, hemolisines and the enzyme activity. The susceptibility tests were performed on Mueller-Hinton medium according to CLSI (Clinical and Laboratory Standards Institute. Results: The strains were: MSSA (methicillin-susceptible S. aureus (74%, MRSA (8%, MLS B (macrolides, lincosamides and type B streptogramines resistance (12% and MRSA and MLS B (6%. MRSA strains were more frequently isolated from sputum. MRSA associated with the MLS B strains were more frequently isolated from pus. MLS B strains were more frequently isolated from sputum and throat secretions. All S. aureus strains were susceptible to vancomycin and teicoplanin. Conclusions: All staphylococcal infections require resistance testing before treatment. MLS B shows a high prevalence among strains of S. aureus. The association between MLS B and MRSA remains a major problem in Romania.

  3. Macrolides resistance of common bacteria isolated from Taiwan.

    Science.gov (United States)

    Chang, S C; Chen, Y C; Luh, K T; Hsieh, W C

    1995-12-01

    To determine the susceptibility to macrolides of common pathogenic bacteria isolated from Taiwan, the in vitro activities of erythromycin, roxithromycin, azithromycin, clarithromycin, and dirithromycin were tested against 492 clinical isolates of eight different bacteria, collected from the National Taiwan University Hospital. The results showed high minimum inhibitory concentrations (MICs) against most of the tested bacteria. The MIC90s for Staphylococcus aureus (both methicillin-resistant and -sensitive strains), coagulase-negative staphylococci (both methicillin-resistant and -sensitive strains), Streptococcus pyogenes, Streptococcus pneumoniae, enterococci, peptostreptococci, and Bacteroides fragilis were all > or = 256 micrograms/ml. The MIC50s for methicillin-resistant strains of S. aureus and coagulase-negative staphylococci, and enterococci were > or = 256 micrograms/ml. For S. pneumoniae, peptostreptococci, and B. fragilis, the MIC50s were > 8 micrograms/ml. The resistance rates to macrolides were 80% or more in methicillin-resistant staphylococci and about 30% in methicillin-sensitive staphylococci. Around 55% of S. pneumoniae strains and 37 approximately 42% of S. pyogenes strains were resistant to macrolides. Cross-resistance to different macrolides was clearly demonstrated in most of the resistant strains.

  4. AMINOGLYCOSIDE RESISTANCE GENES IN Pseudomonas aeruginosa ISOLATES FROM CUMANA, VENEZUELA

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    Bertinellys TEIXEIRA

    2016-01-01

    Full Text Available The enzymatic modification of aminoglycosides by aminoglycoside-acetyltransferases (AAC, aminoglycoside-adenyltransferases (AAD, and aminoglycoside-phosphotransferases (APH, is the most common resistance mechanism in P. aeruginosa and these enzymes can be coded on mobile genetic elements that contribute to their dispersion. One hundred and thirty seven P. aeruginosa isolates from the University Hospital, Cumana, Venezuela (HUAPA were evaluated. Antimicrobial susceptibility was determined by the disk diffusion method and theaac, aadB and aph genes were detected by PCR. Most of the P. aeruginosa isolates (33/137 were identified from the Intensive Care Unit (ICU, mainly from discharges (96/137. The frequency of resistant P. aeruginosaisolates was found to be higher for the aminoglycosides tobramycin and amikacin (30.7 and 29.9%, respectively. Phenotype VI, resistant to these antibiotics, was the most frequent (14/49, followed by phenotype I, resistant to all the aminoglycosides tested (12/49. The aac(6´-Ib,aphA1 and aadB genes were the most frequently detected, and the simultaneous presence of several resistance genes in the same isolate was demonstrated. Aminoglycoside resistance in isolates ofP. aeruginosa at the HUAPA is partly due to the presence of the aac(6´-Ib, aphA1 andaadB genes, but the high rates of antimicrobial resistance suggest the existence of several mechanisms acting together. This is the first report of aminoglycoside resistance genes in Venezuela and one of the few in Latin America.

  5. "DRUG RESISTANCE PATTERN IN ISOLATED BACTERIA FROM BLOOD CULTURES"

    OpenAIRE

    A Sobhani; H. Shodjai S. Javanbakht

    2004-01-01

    Bacteremia is an important infectious disease which may lead to death. Common bacteria and pattern of antibiotic resistance in different communities are different and understanding these differences is important. In the present study, relative frequency and pattern of drug resistance have been examined in bacteria isolated from blood cultures in Razi Hospital laboratory. The method of the study was descriptive. Data collection was carried out retrospectively. Total sample consisted of 311 pos...

  6. Prevalence of Aminoglycoside Resistance Genes in Acinetobacter baumannii Isolates

    OpenAIRE

    Aliakbarzade, Katayun; Farajnia, Safar; Karimi Nik, Ashraf; Zarei, Farzaneh; Tanomand, Asghar

    2014-01-01

    Background: Acinetobacter baumannii is one of the major causes of nosocomial infections and is resistant to most available antibiotics. Aminoglycosides remain as drugs of choice for treatment of Acinetobacter infections yet resistance to aminoglycosides has increased in the recent years. Objectives: The present study investigated the prevalence of genes encoding aminoglycoside-modifying enzymes in A. baumannii strains isolated from patients of Tabriz city, northwest of Iran. Materials and Met...

  7. Expression of anti -apoptotic and multidrug resistance- associated protein genes in glioma stem cell isolated from primary glioma cells%胶质瘤干细胞抗凋亡和多重耐药基因的表达

    Institute of Scientific and Technical Information of China (English)

    陈德勤; 赵洪洋; 靳峰; 赵万巨; 魏宇佳; 张浩; 张军臣; 高超; 郭强; 邵彤

    2010-01-01

    Objective To isolate and culture glioma stem cells from primary glioma cells and detect the expression of anti - apoptotic and multidrug resistance - associated protein (MRP) genes. Methods Glioma stem cells were isolated from human glioma cell by physics technique and nestin, β- tubulin and GFAP expression were examined by Immunofluoresence staining. Expression of livin, livinα, livinβ, survivin, MRPI and MRP3 were assayed by real - time quantitative RT-PCR. Results Glioma stem cells were induced successfully from two glioma specimens from one glioblastoma (No. 912110) and one grade 2 -3 astrocytoma subject (No. 916718). The glioma stem cells which were cultured in NSC medium grew in suspension and presented the features of stem cells: sphere - like shape, self - renewal and ability to differentiate. Nestin immunofluoresence staining of the stem cells presented positive, but the glioma did not show this feature. These cells could conditionally differentiate into tubulin -β~+ and GFAP~+ cells, which provided these cells in culture could produce neurons as well as glial cells. The mRNA expression of cancer stem cells on livin, livinα, survivin, MRP1 was significantly up - regulated than that of glioma primary cells. However, the mRNA expression of MRP3 was down - regulated. Conclusion Primary glioma cell contains cancer stem cells. The expression levels of anti - apeptotic and MRP1 genes are higher than that of glioma primary cells, which show glioma stem cells are the factor to maintain tumor growth and resist apoptosis and to pump the anti - tumor drugs out of cells, and this is what we need further research on.%目的 从人脑胶质瘤组织中分离、培养胶质瘤干细胞,并探讨其生物学特性及其抗凋亡和多重耐药基因的表达差异.方法 人脑胶质瘤组织经过原代细胞培养后,用无血清培养方法获得胶质瘤干细胞球,用10%的胎牛血清培养诱导分化,分化前后分别做nestin、tubulin-β、GFAP免疫细

  8. Ex vivo piperaquine resistance developed rapidly in Plasmodium falciparum isolates in northern Cambodia compared to Thailand

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    Suwanna Chaorattanakawee

    2016-10-01

    Full Text Available Abstract Background The recent dramatic decline in dihydroartemisinin-piperaquine (DHA-PPQ efficacy in northwestern Cambodia has raised concerns about the rapid spread of piperaquine resistance just as DHA-PPQ is being introduced as first-line therapy in neighbouring countries. Methods Ex vivo parasite susceptibilities were tracked to determine the rate of progression of DHA, PPQ and mefloquine (MQ resistance from sentinel sites on the Thai–Cambodian and Thai–Myanmar borders from 2010 to 2015. Immediate ex vivo (IEV histidine-rich protein 2 (HRP-2 assays were used on fresh patient Plasmodium falciparum isolates to determine drug susceptibility profiles. Results IEV HRP-2 assays detected the precipitous emergence of PPQ resistance in Cambodia beginning in 2013 when 40 % of isolates had an IC90 greater than the upper limit of prior years, and this rate doubled to 80 % by 2015. In contrast, Thai–Myanmar isolates from 2013 to 14 remained PPQ-sensitive, while northeastern Thai isolates appeared to have an intermediate resistance profile. The opposite trend was observed for MQ where Cambodian isolates appeared to have a modest increase in overall sensitivity during the same period, with IC50 declining to median levels comparable to those found in Thailand. A significant association between increased PPQ IC50 and IC90 among Cambodian isolates with DHA-PPQ treatment failure was observed. Nearly all Cambodian and Thai isolates were deemed artemisinin resistant with a >1 % survival rate for DHA in the ring-stage assay (RSA, though there was no correlation among isolates to indicate cross-resistance between PPQ and artemisinins. Conclusions Clinical DHA-PPQ failures appear to be associated with declines in the long-acting partner drug PPQ, though sensitivity appears to remain largely intact for now in western Thailand. Rapid progression of PPQ resistance associated with DHA-PPQ treatment failures in northern Cambodia limits drugs of choice in

  9. Isolation, Identification of Heat Resistant Moulds in Margarine and Determination of Their Heat Resistance

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    A. S. Demirci

    2006-09-01

    Full Text Available In this study, moulds that cause problems in a margarine production plant which is located in Trakyaregion have been isolated and identified. In addition to, their heat resistance and lipolytic activity werestudied. For this purpose, margarine samples from various production lots and process water samples fromproduction plant were taken aseptically, transported immediately to the laboratory and analyzed. In thisresearch, two different heat resistant mould strains have been isolated from margarines and process water.After identification of this moulds, their heat resistances at different temperatures have been investigated.Mould isolates were identified as heat resistant Aspergillus fumigatus, Paecilomyces variotii based onmacroscopic and microscopic features. To this analyses results about thermal resistance, Aspergillusfumigatus and Paecilomyces variotii were ability to survive heat treatment at 95oC 10 minutes and 90oC 10minutes, respectively.

  10. Isolation of enamelinlike proteins from blue shark (Prionace glauca) enameloid.

    Science.gov (United States)

    Graham, E E

    1985-05-01

    A sequential dissociative extraction scheme was used to extract proteins from developing Blue Shark enameloid. The first extraction solution (4 M guanidine HC1) solubilized the polypeptides, mainly collagenous, not closely associated with the hydroxyapatite. The next extraction solution (4 M guanidine HC1, 0.5 M ethylenediaminetetraacedic acid (EDTA] solubilized the proteins more closely associated with the tooth mineral component. After extraction, the proteins were separated and isolated with gel electrophoresis. Protein molecular weights were determined and selected proteins were isolated for amino acid composition analysis. The two proteins isolated were tested for mammalian enamel protein antigenic determinants by a "Dot" immunobinding assay. The isolated proteins were enamelinlike by extraction criteria and amino acid composition. Further, the two proteins share antigenic determinants with mammalian enamel proteins.

  11. Emulsion properties of algae soluble protein isolate from Tetraselmis sp.

    NARCIS (Netherlands)

    Schwenzfeier, A.; Helbig, A.; Wierenga, P.A.; Gruppen, H.

    2013-01-01

    To study possible applications of microalgae proteins in foods, a colourless, protein-rich fraction was isolated from Tetraselmis sp. In the present study the emulsion properties of this algae soluble protein isolate (ASPI) were investigated. Droplet size and droplet aggregation of ASPI stabilized

  12. Emulsion properties of algae soluble protein isolate from Tetraselmis sp.

    NARCIS (Netherlands)

    Schwenzfeier, A.; Helbig, A.; Wierenga, P.A.; Gruppen, H.

    2013-01-01

    To study possible applications of microalgae proteins in foods, a colourless, protein-rich fraction was isolated from Tetraselmis sp. In the present study the emulsion properties of this algae soluble protein isolate (ASPI) were investigated. Droplet size and droplet aggregation of ASPI stabilized o

  13. Hydration Properties of Soybean Protein Isolates

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    G. Remondetto

    2001-12-01

    Full Text Available Hydration properties of soybean isolates with different processing conditions (heat treatments, pH and protein concentrations were studied. Degree of denaturation,, solubility in water, in 0.2mol/L NaCl, and in media of different sodium dodecyl sulfate concentrations, viscosity and water imbibing capacity of the different samples were determinated and correlated. Treatments at temperatures higher than 80ºC denatured 11S and 7S proteins, leading to exposure of hydrophobic groups, which produced insoluble aggregates either in water or in high ionic strength media. These isolates possessed high water imbibing capacities and gave rise to viscous dispersions. Significant correlations were obtained between hydration properties and the "m" coefficient as calculated by a power law equation relating viscosity with the protein concentration of the dispersion. This "m" coefficient also correlated with the denaturation enthalpy of the protein isolates. On the basis of these results, it might be suggested that the "m" coefficient - dependent of the hydrodynamic behaviour of the particles - was a good estimator of the degree of protein denaturation.Estudaram-se as propriedades de hidratação de isolados de soja com diferentes condições de procesamento (tratamentos térmicos, pH e concentração de proteínas. Para diferentes amostras determinaram-se e correlacionaram-se o gráu de desnaturalização, a solubilidade em água, em 0,2mol/L NaCl e em diferentes concentrações de dodecil sulfato de sódio, viscosidade e capacidade de absorção de água. Os tratamentos a temperaturas superiores aos 80ºC desnaturaram as fracções 11S e 7S, provocando a exposição de grupos hidrofóbicos os que produziram agregados insolúveis, em água como em solução com alta força iónica. Estes isolados posuiam alta capacidade de absorção de água e dispersões com alta viscosidade. Achou-se uma correlação significativa entre as propriedades de hidratação e o

  14. Functionality of whey protein isolates and hydrolyzed whey proteins

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    Zoran Herceg

    2005-07-01

    Full Text Available The aim of this study was to determine functional properties: solubility, dispersibility, viscosity, emulsifiying, foaming and rheological properties of hydrolyzed whey protein (HWP and whey protein isolate (WPI. Particle size analysis and specific area of HWP and WPI were performed by «Mie – theory» of «light scatering» using «Malvern Mastersizer X». The results of this analysis have shown that HWP had higher particle size and specific area than WPI.By examing functional properties, it has been established that HWP has higher solubility, dispersibility, emulsifiying properties as well as foam stability (FSI and MFS compared to WPI. Rheological properties of protein suspensions were determined in 10, 15, and 18 % suspension of proteins (HWP and WPI by rotational viscosimeter, Brookfiel DV-III at 25°C. Apparent viscosity at 200 s-1 was calculated, using Newtonian law. On the basis of measured shear rate and shear stress, using least squere method, rheological parameters, flow behavior indeks (n and consistency coefficient (k were determined according to the Ostwald de Waele model. The highest viscosity was observed in 18% HWP model system while the least viscosity was found in a model system prepared with 10% WPI.

  15. Antimicrobial resistance of bacterial enteropathogens isolated from stools in Madagascar.

    Science.gov (United States)

    Randrianirina, Frederique; Ratsima, Elisoa Hariniana; Ramparany, Lova; Randremanana, Rindra; Rakotonirina, Hanitra Clara; Andriamanantena, Tahiry; Rakotomanana, Fanjasoa; Rajatonirina, Soatiana; Richard, Vincent; Talarmin, Antoine

    2014-02-25

    Diarrheal diseases are a major public health problem in developing countries, and are one of the main causes of hospital admissions in Madagascar. The Pasteur Institute of Madagascar undertook a study to determine the prevalence and the pathogenicity of bacterial, viral and protozoal enteropathogens in diarrheal and non-diarrheal stools of children aged less than 5 years in Madagascar. We present here the results of the analysis of antimicrobial susceptibility of the bacteria isolated during this study. The study was conducted in the community setting in 14 districts of Madagascar from October 2008 to May 2009. Conventional methods and PCR were used to identify the bacteria; antimicrobial susceptibility was determined using an agar diffusion method for enterobacteriaceae and MICs were measured by an agar dilution method for Campylobacter sp. In addition to the strains isolated during this study, Salmonella sp and Shigella sp isolated at the Pasteur Institute of Madagascar from 2005 to 2009 were included in the analysis to increase the power of the study. Twenty-nine strains of Salmonella sp, 35 strains of Shigella sp, 195 strains of diarrheagenic E. coli, 203 strains of C. jejuni and 71 strains of C. coli isolated in the community setting were tested for antibiotic resistance. Fifty-five strains of Salmonella sp and 129 strains of Shigella sp isolated from patients referred to the Pasteur Institute of Madagascar were also included in the study. Many E. coli and Shigella isolates (around 80%) but fewer Salmonella isolates were resistant to ampicillin and trimethoprim/sulfamethoxazole. A small proportion of strains of each species were resistant to ciprofloxacin and only 3% of E. coli strains presented a resistance to third generation cephalosporins due to the production of extended-spectrum beta-lactamases. The resistance of Campylobacter sp to ampicillin was the most prevalent, whereas less than 5% of isolates were resistant to each of the other antibiotics. The

  16. Molecular characterization of multidrug-resistant Klebsiella pneumoniae isolates.

    Science.gov (United States)

    Hou, Xiang-hua; Song, Xiu-yu; Ma, Xiao-bo; Zhang, Shi-yang; Zhang, Jia-qin

    2015-01-01

    Klebsiella pneumoniae is an important cause of healthcare-associated infections worldwide. Selective pressure, the extensive use of antibiotics, and the conjugational transmission of antibiotic resistance genes across bacterial species and genera facilitate the emergence of multidrug-resistant (MDR) K. pneumoniae. Here, we examined the occurrence, phenotypes and genetic features of MDR K. pneumoniae isolated from patients in intensive care units (ICUs) at the First Affiliated Hospital of Xiamen University in Xiamen, China, from January to December 2011. Thirty-eight MDR K. pneumoniae strains were collected. These MDR K. pneumoniae isolates possessed at least seven antibiotic resistance determinants, which contribute to the high-level resistance of these bacteria to aminoglycosides, macrolides, quinolones and β-lactams. Among these isolates, 24 strains were extended-spectrum β-lactamase (ESBL) producers, 2 strains were AmpC producers, and 12 strains were both ESBL and AmpC producers. The 38 MDR isolates also contained class I (28/38) and class II integrons (10/38). All 28 class I-positive isolates contained aacC1, aacC4, orfX, orfX' and aadA1 genes. β-lactam resistance was conferred through bla SHV (22/38), bla TEM (10/38), and bla CTX-M (7/38). The highly conserved bla KPC-2 (37/38) and bla OXA-23(1/38) alleles were responsible for carbapenem resistance, and a gyrAsite mutation (27/38) and the plasmid-mediated qnrB gene (13/38) were responsible for quinolone resistance. Repetitive-sequence-based PCR (REP-PCR) fingerprinting of these MDR strains revealed the presence of five groups and sixteen patterns. The MDR strains from unrelated groups showed different drug resistance patterns; however, some homologous strains also showed different drug resistance profiles. Therefore, REP-PCR-based analyses can provide information to evaluate the epidemic status of nosocomial infection caused by MDR K. pneumoniae; however, this test lacks the power to discriminate some

  17. Molecular characterization of multidrug-resistant Klebsiella pneumoniae isolates

    Directory of Open Access Journals (Sweden)

    Xiang-hua Hou

    2015-09-01

    Full Text Available Klebsiella pneumoniae is an important cause of healthcare-associated infections worldwide. Selective pressure, the extensive use of antibiotics, and the conjugational transmission of antibiotic resistance genes across bacterial species and genera facilitate the emergence of multidrug-resistant (MDR K. pneumoniae. Here, we examined the occurrence, phenotypes and genetic features of MDR K. pneumoniae isolated from patients in intensive care units (ICUs at the First Affiliated Hospital of Xiamen University in Xiamen, China, from January to December 2011. Thirty-eight MDR K. pneumoniae strains were collected. These MDR K. pneumoniae isolates possessed at least seven antibiotic resistance determinants, which contribute to the high-level resistance of these bacteria to aminoglycosides, macrolides, quinolones and β-lactams. Among these isolates, 24 strains were extended-spectrum β-lactamase (ESBL producers, 2 strains were AmpC producers, and 12 strains were both ESBL and AmpC producers. The 38 MDR isolates also contained class I (28/38 and class II integrons (10/38. All 28 class I-positive isolates contained aacC1, aacC4, orfX, orfX’ and aadA1 genes. β-lactam resistance was conferred through blaSHV (22/38, blaTEM (10/38, and blaCTX-M (7/38. The highly conserved blaKPC-2 (37/38 and blaOXA-23(1/38 alleles were responsible for carbapenem resistance, and a gyrAsite mutation (27/38 and the plasmid-mediated qnrB gene (13/38 were responsible for quinolone resistance. Repetitive-sequence-based PCR (REP-PCR fingerprinting of these MDR strains revealed the presence of five groups and sixteen patterns. The MDR strains from unrelated groups showed different drug resistance patterns; however, some homologous strains also showed different drug resistance profiles. Therefore, REP-PCR-based analyses can provide information to evaluate the epidemic status of nosocomial infection caused by MDR K. pneumoniae; however, this test lacks the power to discriminate some

  18. Identification of Functional Regulatory Residues of the β-Lactam Inducible Penicillin Binding Protein in Methicillin-Resistant Staphylococcus aureus

    OpenAIRE

    Mbah, Andreas N.; Isokpehi, Raphael D

    2013-01-01

    Resistance to methicillin by Staphylococcus aureus is a persistent clinical problem worldwide. A mechanism for resistance has been proposed in which methicillin resistant Staphylococcus aureus (MRSA) isolates acquired a new protein called β -lactam inducible penicillin binding protein (PBP-2′). The PBP-2′ functions by substituting other penicillin binding proteins which have been inhibited by β -lactam antibiotics. Presently, there is no structural and regulatory information on PBP-2′ protein...

  19. New Ceftriaxone- and Multidrug-Resistant Neisseria gonorrhoeae Strain with a Novel Mosaic penA Gene Isolated in Japan.

    Science.gov (United States)

    Nakayama, Shu-Ichi; Shimuta, Ken; Furubayashi, Kei-Ichi; Kawahata, Takuya; Unemo, Magnus; Ohnishi, Makoto

    2016-07-01

    We have characterized in detail a new ceftriaxone- and multidrug-resistant Neisseria gonorrhoeae strain (FC428) isolated in Japan in 2015. FC428 differed from previous ceftriaxone-resistant strains and contained a novel mosaic penA allele encoding a new mosaic penicillin-binding protein 2 (PBP 2). However, the resistance-determining 3'-terminal region of penA was almost identical to the regions of two previously reported ceftriaxone-resistant strains from Australia and Japan, indicating that both ceftriaxone-resistant strains and conserved ceftriaxone resistance-determining PBP 2 regions might spread.

  20. Antibiotic Resistance Patterns in Invasive Group B Streptococcal Isolates

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    Mei L. Castor

    2008-01-01

    Full Text Available Antibiotics are used for both group B streptococcal (GBS prevention and treatment. Active population-based surveillance for invasive GBS disease was conducted in four states during 1996—2003. Of 3813 case-isolates, 91.0% (3471 were serotyped, 77.1% (2937 had susceptibility testing, and 46.6% (3471 had both. All were sensitive to penicillin, ampicillin, cefazolin, cefotaxime, and vancomycin. Clindamycin and erythromycin resistance was 12.7% and 25.6%, respectively, and associated with serotype V (P<.001. Clindamycin resistance increased from 10.5% to 15.0% (X2 for trend 12.70; P<.001; inducible clindamycin resistance was associated with the erm genotype. Erythromycin resistance increased from 15.8% to 32.8% (X2 for trend 55.46; P<.001. While GBS remains susceptible to beta-lactams, resistance to alternative agents such as erythromycin and clindamycin is an increasing concern.

  1. Isolated cell behavior drives the evolution of antibiotic resistance.

    Science.gov (United States)

    Artemova, Tatiana; Gerardin, Ylaine; Dudley, Carmel; Vega, Nicole M; Gore, Jeff

    2015-07-29

    Bacterial antibiotic resistance is typically quantified by the minimum inhibitory concentration (MIC), which is defined as the minimal concentration of antibiotic that inhibits bacterial growth starting from a standard cell density. However, when antibiotic resistance is mediated by degradation, the collective inactivation of antibiotic by the bacterial population can cause the measured MIC to depend strongly on the initial cell density. In cases where this inoculum effect is strong, the relationship between MIC and bacterial fitness in the antibiotic is not well defined. Here, we demonstrate that the resistance of a single, isolated cell-which we call the single-cell MIC (scMIC)-provides a superior metric for quantifying antibiotic resistance. Unlike the MIC, we find that the scMIC predicts the direction of selection and also specifies the antibiotic concentration at which selection begins to favor new mutants. Understanding the cooperative nature of bacterial growth in antibiotics is therefore essential in predicting the evolution of antibiotic resistance.

  2. Bipolar resistive switching in different plant and animal proteins

    KAUST Repository

    Bag, A.

    2014-06-01

    We report bipolar resistive switching phenomena observed in different types of plant and animal proteins. Using protein as the switching medium, resistive switching devices have been fabricated with conducting indium tin oxide (ITO) and Al as bottom and top electrodes, respectively. A clockwise bipolar resistive switching phenomenon is observed in all proteins. It is shown that the resistive switching phenomena originate from the local redox process in the protein and the ion exchange from the top electrode/protein interface.

  3. Isolation, identification and antibiotic resistance of Campylobacter strains isolated from domestic and free-living pigeons.

    Science.gov (United States)

    Dudzic, A; Urban-Chmiel, R; Stępień-Pyśniak, D; Dec, M; Puchalski, A; Wernicki, A

    2016-04-01

    1. The aim of this study was to evaluate the occurrence of Campylobacter spp. in domestic and free-living pigeons and to evaluate the antibiotic resistance profiles. 2. The material consisted of cloacal swabs obtained from 108 homing pigeons and fresh faeces from 72 wild birds from Lublin and its vicinity. The identification of strains isolated on differential/selective media for Campylobacter spp. was carried out by MALDI-TOF and PCR. The susceptibility to antibiotics was evaluated by minimum inhibitory concentration (MIC) in Mueller-Hinton broth. 3. A total of 35 strains of Campylobacter spp. were isolated; 27 were identified as Campylobacter jejuni and 8 as Campylobacter coli. Over half of the isolates were resistant to erythromycin and streptomycin, 40% of strains were resistant to tetracycline and ampicillin and 37% isolates were resistant to amoxicillin. Resistance to two or more antibiotics was observed in all strains tested. 4. The results indicate that both domestic and free-living pigeons are reservoirs for bacteria of the genus Campylobacter, which are characterised by varied and growing resistance to commonly used antibiotics.

  4. Predicting Resistance Mutations Using Protein Design Algorithms

    Energy Technology Data Exchange (ETDEWEB)

    Frey, K.; Georgiev, I; Donald, B; Anderson, A

    2010-01-01

    Drug resistance resulting from mutations to the target is an unfortunate common phenomenon that limits the lifetime of many of the most successful drugs. In contrast to the investigation of mutations after clinical exposure, it would be powerful to be able to incorporate strategies early in the development process to predict and overcome the effects of possible resistance mutations. Here we present a unique prospective application of an ensemble-based protein design algorithm, K*, to predict potential resistance mutations in dihydrofolate reductase from Staphylococcus aureus using positive design to maintain catalytic function and negative design to interfere with binding of a lead inhibitor. Enzyme inhibition assays show that three of the four highly-ranked predicted mutants are active yet display lower affinity (18-, 9-, and 13-fold) for the inhibitor. A crystal structure of the top-ranked mutant enzyme validates the predicted conformations of the mutated residues and the structural basis of the loss of potency. The use of protein design algorithms to predict resistance mutations could be incorporated in a lead design strategy against any target that is susceptible to mutational resistance.

  5. Mutations in transcription factor Mrr2p contribute to fluconazole resistance in clinical isolates of Candida albicans.

    Science.gov (United States)

    Wang, Ying; Liu, Jin-Yan; Shi, Ce; Li, Wen-Jing; Zhao, Yue; Yan, Lan; Xiang, Ming-Jie

    2015-11-01

    The Candida albicans zinc cluster proteins are a family of transcription factors (TFs) that play essential roles in the development of antifungal drug resistance. Gain-of-function mutations in several TFs, such as Tac1p, Mrr1p and Upc2p, have been previously well documented in azole-resistant clinical C. albicans isolates. Mrr2p (multidrug resistance regulator 2) is a novel TF controlling expression of the ABC transporter gene CDR1 and mediating fluconazole resistance. In this study, the relationship between naturally occurring mutations in MRR2 and fluconazole resistance in clinical C. albicans isolates was investigated. Among a group of 20 fluconazole-resistant clinical C. albicans and 10 fluconazole-susceptible C. albicans, 12 fluconazole-resistant isolates overexpressed CDR1 by at least two-fold compared with the fluconazole-susceptible isolates. Of these 12 resistant isolates, three (C7, C9, C15) contained 11 identical missense mutations, 6 of which occurred only in the azole-resistant isolates. The contribution of these mutations to CDR1 overexpression and therefore to fluconazole resistance was further verified by generating recombinant strains containing the mutated MRR2 gene. The mutated MRR2 alleles from isolate C9 contributed to an almost six-fold increase in CDR1 expression and an eight-fold increase in fluconazole resistance; the missense mutations S466L and T470N resulted in an increase in CDR1 expression of more than two-fold and a four-fold increase in fluconazole resistance. In contrast, the other four missense mutations conferred only two- to four-fold increases in fluconazole resistance, with no significant increase in CDR1 expression. These findings provide some insight into the mechanism by which MRR2 regulates C. albicans multidrug resistance.

  6. A novel multidrug resistance plasmid isolated from an Escherichia coli strain resistant to aminoglycosides.

    Science.gov (United States)

    Sun, Hui; Li, Shasha; Xie, Zhijing; Yang, Fangfang; Sun, Yani; Zhu, Yanli; Zhao, Xiaomin; Jiang, Shijin

    2012-07-01

    Previous studies have reported several different plasmids that confer multidrug resistance (MDR) including resistance to aminoglycosides. In this study, we investigated the aminoglycoside resistance patterns for 224 Escherichia coli isolates from diseased chickens and ducks in China, characterized a novel MDR plasmid, and collected prevalence data on similar resistance plasmids. Antibiotic susceptibilities were determined using disc diffusion and the microdilution method. The plasmid pXZ was analysed by restriction fragment length polymorphism (RFLP) with EcoRI and SalI, and sequenced. The prevalence of similar resistance plasmids was assessed by multiplex PCR and by RFLP analysis. Among the 224 E. coli isolates, 189 (84.4%) were resistant to streptomycin, 125 (55.8%) were resistant to kanamycin, 116 (51.8%) were resistant to gentamicin, 106 (47.3%) were resistant to neomycin and 98 (43.8%) were resistant to amikacin. Among the 224 E. coli isolates, 17 contained a plasmid with the MDR-encoding region of pXZ, which showed high-level resistance to aminoglycosides (MICs of gentamicin and amikacin ≥ 512 mg/L). The plasmid pXZ was digested into five fragments by EcoRI and six fragments by SalI. The plasmid pXZ was a circular DNA molecule of 76635 bp with a 51.65% guanine + cytosine content and included four resistance genes (rmtB, fosA3, bla(TEM-1) and bla(CTX-M-24)). A novel MDR plasmid, pXZ, harbouring four resistance genes (rmtB, fosA3, bla(TEM-1) and bla(CTX-M)) was identified. To our knowledge, this is the first report of an aminoglycoside resistance plasmid harbouring the fosA3 gene.

  7. Antimicrobial resistance determinant microarray for analysis of multi-drug resistant isolates

    Science.gov (United States)

    Taitt, Chris Rowe; Leski, Tomasz; Stenger, David; Vora, Gary J.; House, Brent; Nicklasson, Matilda; Pimentel, Guillermo; Zurawski, Daniel V.; Kirkup, Benjamin C.; Craft, David; Waterman, Paige E.; Lesho, Emil P.; Bangurae, Umaru; Ansumana, Rashid

    2012-06-01

    The prevalence of multidrug-resistant infections in personnel wounded in Iraq and Afghanistan has made it challenging for physicians to choose effective therapeutics in a timely fashion. To address the challenge of identifying the potential for drug resistance, we have developed the Antimicrobial Resistance Determinant Microarray (ARDM) to provide DNAbased analysis for over 250 resistance genes covering 12 classes of antibiotics. Over 70 drug-resistant bacteria from different geographic regions have been analyzed on ARDM, with significant differences in patterns of resistance identified: genes for resistance to sulfonamides, trimethoprim, chloramphenicol, rifampin, and macrolide-lincosamidesulfonamide drugs were more frequently identified in isolates from sources in Iraq/Afghanistan. Of particular concern was the presence of genes responsible for resistance to many of the last-resort antibiotics used to treat war traumaassociated infections.

  8. Antibiotic Resistance of Vibrio cholerae Isolates from Kashan, Iran

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    Afzali H.MD,

    2016-03-01

    Full Text Available Abstract Aims: Cholera is an acute diarrheal disease that can lead to severe dehydration and death. Antibiotic resistance is a big challenge in infective disease like Cholera. The present study aimed to understand the characteristics and trends of antibiotic resistance of V. cholerae isolations in and around Kashan, Iran. Instrument & Methods: In this descriptive cross-sectional study, samples were gathered using census method from 1998 to 2013 in Kashan, Iran. 1132 fecal samples of patients with acute diarrhea and 237 samples of suspected water samples were taken. The serotypes and biotypes were determined by an enzymatic method. Antibiotic susceptibility test was performed by using Disk Diffusion Method. Data were analyzed using SPSS 23 software. Fisher-exact and Chi-square tests were used to compare the statistical parameters. Findings: 96 fecal samples (8.5% and 18 water samples (7.6% were positive for Vibrio cholerae. Non-agglutinating (Nag isolates (75.4% were more common than serotype Inaba (13.2% and Ogawa (11.4%. Nag serotypes were mostly resistant to cefixime (44% and ampicillin (33%. In contaminated water samples also the most frequent cases were Nag serotype (50%. Nag serotype showed 22.2% of resistance to ampicillin and nitrofurantoin. Conclusion: Vibrio cholerae isolates in Kashan, Iran, are highly resistant to antibiotics, especially Nag serotypes.

  9. METRONIDAZOLE RESISTANCE IN ANAEROBES ISOLATED FROM CHRONIC PERIODONTITIS CASES

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    Sushma Narayan

    2016-01-01

    Full Text Available Periodontitis is the most frequent oral health problem in the world. The infection is primarily caused by anaerobic microorganisms. Metronidazole is the most commonly used drug to treat the infection but recently the anaerobes have shown the resistance to this drug. Therefore, the present study was undertaken to isolate and identify the anaerobes associated with periodontitis and study their susceptibility pattern to the Metronidazole. Total 90 samples were collected from chronic periodontitis cases. Anaerobes were isolated in 71% of periodontitis cases. Gram positive organisms were more predominantly isolated than Gram negative organisms. The Gram negative anaerobes were found to be 100% sensitive to Metronidazole while Gram positive anaerobes showed 8% resistance to Metronidazole.

  10. Resisting protein adsorption on biodegradable polyester brushes.

    Science.gov (United States)

    Hu, Xinfang; Gorman, Christopher B

    2014-08-01

    The protein adsorption and degradation behaviors of poly(lactic acid), poly(glycolic acid) (PGA) and poly(ε-caprolactone) (PCL) brushes and their co-polymer brushes with oligo(ethylene glycol) (OEG) were studied. Both brush structure and relative amount of OEG and polyester were found to be important to the protein resistance of the brushes. A protein-resisting surface can be fabricated either by using OEG as the top layer of a copolymer brush or by increasing the amount of OEG relative to polyester when using a hydroxyl terminated OEG (OEG-OH) and a methoxy terminated OEG (OEG-OMe) mixture as the substrate layer. The degradation of single polyester brushes and their co-polymer brushes using OEG-OH as a substrate layer or using OEG as a top layer was hindered. This phenomenon was rationalized by the inhibition of the proposed back-biting process as the hydroxy end groups of polyester were blocked by OEG molecules. Among these brushes tested, PGA co-polymer brushes using the methoxy/hydroxyl OEG mixture as the substrate layer proved to be both protein-resistant and degradable due to the relatively large amount of OEG moieties and the good biodegradability of PGA.

  11. Ampicillin-resistant Haemophilus influenzae isolates in Geneva: serotype, antimicrobial susceptibility, and β-lactam resistance mechanisms.

    Science.gov (United States)

    Cherkaoui, A; Diene, S M; Emonet, S; Renzi, G; Francois, P; Schrenzel, J

    2015-10-01

    The purpose of this study was to analyze the molecular mechanisms of ampicillin-resistant Haemophilus influenzae isolated in Geneva, Switzerland. We investigated the association between specific patterns of amino acid substitutions in penicillin-binding protein 3 (with or without β-lactamase production) and β-lactam susceptibility. Another main focus for this study was to compare the accuracy of disk diffusion and Etest methods to detect resistance to ampicillin and amoxicillin/clavulanic acid. The antibiotic susceptibility to β-lactam antibiotics of 124 H. influenzae isolates was determined by disk diffusion and Etest methods, and interpreted by European Committee on Antimicrobial Susceptibility Testing (EUCAST) and Clinical and Laboratory Standards Institute (CLSI) breakpoints. Alterations in PBP3 were investigated by sequencing the ftsI gene. Of the 124 clinical isolates analyzed, ampicillin resistance was found in 36% (45 out of 124). The rate of resistance to amoxicillin/clavulanic acid was 9% and 0.8%, using EUCAST and CLSI breakpoints respectively. For the 78 β-lactamase negative ampicillin-susceptible (BLNAS) isolates for which the Etest method indicated a high degree of susceptibility (MIC ≤ 1 mg/L), the disk diffusion method revealed resistance to ampicillin and amoxicillin/clavulanic acid in 33 cases (42%). Most common amino acid substitutions were Asn526Lys and Val547Ile, followed by Asp569Ser, Ala502Val, Asp350Asn, Met377Ile, Ile449Val, and Arg517His. The patterns observed were classified into six groups (IIa, IIb, IIc, IId, III-like, and miscellaneous). Continued characterization of both invasive and respiratory H. influenzae isolates is necessary in order to observe changes in the microbiology and epidemiology of this pathogen that could lead to clinical failure when treated by empirical antibiotic therapy.

  12. Exploring the contribution of efflux on the resistance to fluoroquinolones in clinical isolates of Escherichia coli.

    Science.gov (United States)

    Paltansing, Sunita; Tengeler, Anouk C; Kraakman, Margriet E M; Claas, Eric C J; Bernards, Alexandra T

    2013-12-01

    Resistance to ciprofloxacin in Escherichia coli is increasing parallel to increased use of fluoroquinolones both in The Netherlands and in other European countries. The objective was to investigate the contribution of active efflux and expression of outer membrane proteins (OMPs) in a collection of clinical E. coli isolates collected at a clinical microbiology department in a Dutch hospital. Forty-seven E. coli isolates a wide range of ciprofloxacin minimum inhibitory concentrations and known mutations in the quinolone resistance determining region were included. A fluorometric determination of bisbenzimide efflux was used two different efflux pump inhibitors and compared to quantitative reverse transcription-polymerase chain reaction (qRT-PCR) for the expression levels of acrA, acrB, tolC, yhiV, and mdfA efflux pump genes and the OMPs ompF and ompX. Six isolates (12.7%) showed increased efflux. Although in 35 isolates (76%), overexpression of ≥1 efflux pump genes using qRT-PCR was present. Only the combined overexpression of acrAB-TolC and mdfA correlated with the phenotypic efflux assay using glucose/carbonyl cyanide m-chlorophenylhydrazone with glucose. Thus, efflux was involved in ciprofloxacin resistance in a limited number of E. coli isolates collected at a clinical microbiology department in a Dutch hospital complementing other resistance mechanisms.

  13. Proteomic analysis of a NAP1 Clostridium difficile clinical isolate resistant to metronidazole.

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    Patrick M Chong

    Full Text Available BACKGROUND: Clostridium difficile is an anaerobic, Gram-positive bacterium that has been implicated as the leading cause of antibiotic-associated diarrhea. Metronidazole is currently the first-line treatment for mild to moderate C. difficile infections. Our laboratory isolated a strain of C. difficile with a stable resistance phenotype to metronidazole. A shotgun proteomics approach was used to compare differences in the proteomes of metronidazole-resistant and -susceptible isolates. METHODOLOGY/PRINCIPAL FINDINGS: NAP1 C. difficile strains CD26A54_R (Met-resistant, CD26A54_S (reduced- susceptibility, and VLOO13 (Met-susceptible were grown to mid-log phase, and spiked with metronidazole at concentrations 2 doubling dilutions below the MIC. Peptides from each sample were labeled with iTRAQ and subjected to 2D-LC-MS/MS analysis. In the absence of metronidazole, higher expression was observed of some proteins in C. difficile strains CD26A54_S and CD26A54_R that may be involved with reduced susceptibility or resistance to metronidazole, including DNA repair proteins, putative nitroreductases, and the ferric uptake regulator (Fur. After treatment with metronidazole, moderate increases were seen in the expression of stress-related proteins in all strains. A moderate increase was also observed in the expression of the DNA repair protein RecA in CD26A54_R. CONCLUSIONS/SIGNIFICANCE: This study provided an in-depth proteomic analysis of a stable, metronidazole-resistant C. difficile isolate. The results suggested that a multi-factorial response may be associated with high level metronidazole-resistance in C. difficile, including the possible roles of altered iron metabolism and/or DNA repair.

  14. Isolation and characterization of a new Cu-Fe protein from Desulfovibrio aminophilus DSM12254.

    Science.gov (United States)

    Rivas, Maria G; Mota, Cristiano S; Pauleta, Sofia R; Carepo, Marta S P; Folgosa, Filipe; Andrade, Susana L A; Fauque, Guy; Pereira, Alice S; Tavares, Pedro; Calvete, Juan J; Moura, Isabel; Moura, José J G

    2009-10-01

    The isolation and characterization of a new metalloprotein containing Cu and Fe atoms is reported. The as-isolated Cu-Fe protein shows an UV-visible spectrum with absorption bands at 320 nm, 409 nm and 615 nm. Molecular mass of the native protein along with denaturating electrophoresis and mass spectrometry data show that this protein is a multimer consisting of 14+/-1 subunits of 15254.3+/-7.6 Da. Mössbauer spectroscopy data of the as-isolated Cu-Fe protein is consistent with the presence of [2Fe-2S](2+) centers. Data interpretation of the dithionite reduced protein suggest that the metallic cluster could be constituted by two ferromagnetically coupled [2Fe-2S](+) spin delocalized pairs. The biochemical properties of the Cu-Fe protein are similar to the recently reported molybdenum resistance associated protein from Desulfovibrio, D. alaskensis. Furthermore, a BLAST search from the DNA deduced amino acid sequence shows that the Cu-Fe protein has homology with proteins annotated as zinc resistance associated proteins from Desulfovibrio, D. alaskensis, D. vulgaris Hildenborough, D. piger ATCC 29098. These facts suggest a possible role of the Cu-Fe protein in metal tolerance.

  15. Cdr2p contributes to fluconazole resistance in Candida dubliniensis clinical isolates.

    LENUS (Irish Health Repository)

    2011-05-01

    The development of resistance to azole antifungals used in the treatment of fungal infections can be a serious medical problem. Here, we investigate the molecular mechanisms associated with reduced susceptibility to fluconazole in clinical isolates of Candida dubliniensis , showing evidence of the trailing growth phenomenon. The changes in membrane sterol composition were studied in the presence of subinhibitory fluconazole concentrations. Despite lanosterol and eburicol accumulating as the most prevalent sterols after fluconazole treatment, these ergosterol precursors still support growth of Candida isolates. The overexpression of ABC transporters was demonstrated by immunoblotting employing specific antibodies against Cdr1p and Cdr2p. The presence of a full-length 170 kDa protein Cdr1p was detected in two isolates, while a truncated form of Cdr1p with the molecular mass of 85 kDa was observed in isolate 966\\/3(2). Notably, Cdr2p was detected in this isolate, and the expression of this transporter was modulated by subinhibitory concentrations of fluconazole. These results suggest that C. dubliniensis can display the trailing growth phenomenon, and such isolates express similar molecular mechanisms like that of fluconazole-resistant isolates and can therefore be associated with recurrent infections.

  16. 21 CFR 172.340 - Fish protein isolate.

    Science.gov (United States)

    2010-04-01

    ..., as determined by the method described in section 2.057, Improved Kjeldahl Method for Nitrate-Free... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Fish protein isolate. 172.340 Section 172.340 Food... Special Dietary and Nutritional Additives § 172.340 Fish protein isolate. (a) The food additive...

  17. "DRUG RESISTANCE PATTERN IN ISOLATED BACTERIA FROM BLOOD CULTURES"

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    A. Sobhani

    2004-05-01

    Full Text Available Bacteremia is an important infectious disease which may lead to death. Common bacteria and pattern of antibiotic resistance in different communities are different and understanding these differences is important. In the present study, relative frequency and pattern of drug resistance have been examined in bacteria isolated from blood cultures in Razi Hospital laboratory. The method of the study was descriptive. Data collection was carried out retrospectively. Total sample consisted of 311 positive blood cultures from 1999 to 2001. Variables under study were bacterial strains, antibiotics examined in antibiogram, microbial resistance, and patients' age and sex. The most common isolated bacteria were Salmonella typhi (22.2% and the least common ones were Citrobacter (1.6%. The highest antibiotic resistance was seen against amoxicillin (88.4%. The proportion of males to females was1: 1/1 and the most common age group was 15-44 (47.3%. Common bacteria and pattern of antibiotic resistance were different in some areas and this subject requires further studies in the future.

  18. Activity of ceftobiprole against Streptococcus pneumoniae isolates exhibiting high-level resistance to ceftriaxone.

    Science.gov (United States)

    Davies, Todd A; Flamm, Robert K; Lynch, A Simon

    2012-06-01

    Tracking Resistance in the US Today (TRUST) 2008 surveillance data showed that 6% of Streptococcus pneumoniae were non-susceptible to ceftriaxone [minimum inhibitory concentration (MIC) ≥ 2 μg/mL] and that 8% of the ceftriaxone-non-susceptible isolates exhibited high-level resistance (MIC ≥ 8 μg/mL). Here we describe the activity of ceftobiprole against ceftriaxone-resistant isolates and characterise the genotypic traits associated with resistance. Thirty isolates with ceftriaxone MICs ≥ 8 μg/mL were analysed by sequencing of penicillin-binding protein (PBP) and murM genes. Sequencing of pbp1a, pbp2b and pbp2x showed nine PBP patterns, with the most common (n=17) being: PBP1a T371S (STMK motif), P432T (SRNVP motif); PBP2b T446A (SSNT motif), A619G (KTGTA motif); and PBP2x T338A and M339F (STMK motif), L364F, I371T, R384G, M400T, L546V (LKSGT motif); six isolates had the same pattern without the PBP2b A619G change. For these 23 isolates, MICs were 8 μg/mL for ceftriaxone, 4-8 μg/mL for penicillin and 0.5-2 μg/mL for ceftobiprole. The remaining seven isolates with higher MICs (ceftriaxone 8-32 μg/mL, penicillin 4-32 μg/mL and ceftobiprole 2-4 μg/mL) had fewer PBP active-site motif substitutions. The majority of isolates (17/30) had murM alleles similar to the wild-type, whilst the rest had alleles reflecting a mosaic structure. No murM alleles were associated with higher MICs. Against these 30 isolates, ceftobiprole was 4-16-fold more active than ceftriaxone. Widely described PBP and MurM substitutions probably account for the high ceftriaxone MICs (8 μg/mL) in the majority of isolates. However, seven isolates with ceftriaxone MICs of 8-32 μg/mL had fewer PBP substitutions in active-site motifs, suggesting either that there is another resistance mechanism or that unique PBP mutations may contribute to high-level β-lactam resistance.

  19. Screening for streptomycin resistance-conferring mutations in Mycobacterium tuberculosis clinical isolates from Poland.

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    Tomasz Jagielski

    Full Text Available Currently, mutations in three genes, namely rrs, rpsL, and gidB, encoding 16S rRNA, ribosomal protein S12, and 16S rRNA-specific methyltransferase, respectively, are considered to be involved in conferring resistance to streptomycin (STR in Mycobacterium tuberculosis. The aim of this study was to investigate the spectrum and frequency of these mutations in M. tuberculosis clinical isolates, both resistant and susceptible to STR. Sixty-four M. tuberculosis isolates recovered from as many TB patients from Poland in 2004 were included in the study. Within the sample were 50 multidrug-resistant (32 STR-resistant and 18 STR-susceptible and 14 pan-susceptible isolates. Preliminary testing for STR resistance was performed with the 1% proportion method. The MICs of STR were determined by the Etest method. Mutation profiling was carried out by amplifying and sequencing the entire rrs, rpsL, and gidB genes. Non-synonymous mutations in either rrs or rpsL gene were detected in 23 (71.9% of the STR-resistant and none of the STR-susceptible isolates. Mutations in the gidB gene were distributed among 12 (37.5% STR-resistant and 13 (40.6% STR-susceptible isolates. Four (12.5% STR-resistant isolates were wild-type at all three loci examined. None of the rrs, rpsL or gidB mutations could be linked to low, intermediate or high level of STR resistance. In accordance with previous findings, the gidB 47T→G (L16R mutation was associated with the Latin American-Mediterranean genotype family, whereas 276A→C (E92D and 615A→G (A205A mutations of the gidB gene were associated with the Beijing lineage. The study underlines the usefulness of rrs and rpsL mutations as molecular markers for STR resistance yet not indicative of its level. The gidB polymorphisms can serve as phylogenetic markers.

  20. Nutritional and functional properties of whey proteins concentrate and isolate

    Directory of Open Access Journals (Sweden)

    Zoran Herceg

    2006-12-01

    Full Text Available Whey protein fractions represent 18 - 20 % of total milk nitrogen content. Nutritional value in addition to diverse physico - chemical and functional properties make whey proteins highly suitable for application in foodstuffs. In the most cases, whey proteins are used because of their functional properties. Whey proteins possess favourable functional characteristics such as gelling, water binding, emulsification and foaming ability. Due to application of new process techniques (membrane fractionation techniques, it is possible to produce various whey - protein based products. The most important products based on the whey proteins are whey protein concentrates (WPC and whey protein isolates (WPI. The aim of this paper was to give comprehensive review of nutritional and functional properties of the most common used whey proteins (whey protein concentrate - WPC and whey protein isolate - WPI in the food industry.

  1. A cfr-positive clinical staphylococcal isolate from India with multiple mechanisms of linezolid-resistance

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    Vineeth Rajan

    2014-01-01

    Full Text Available Background & objectives: Linezolid, a member of the oxazolidinone class of antibiotics, has been an effective therapeutic option to treat severe infections caused by multidrug resistant Gram positive bacteria. Emergence of linezolid resistant clinical strains is a serious issue in the healthcare settings worldwide. We report here the molecular characterization of a linezolid resistant clinical isolate of Staphylococcus haemolyticus from India. Methods: The species of the clinical isolate was identified by 16S rRNA gene sequencing. The minimum inhibitory concentrations (MICs of linezolid, clindamycin, chloramphenicol and oxacillin were determined by E-test method. To elucidate the mechanism of linezolid-resistance, presence of cfr gene (chloramphenicol florfenicol resistance and mutations in 23S rRNA and ribosomal proteins (L3, L4 and L22 were investigated. Staphylococcal Cassette Chromosome mec (SCCmec typing was performed by multiplex PCR. Results: The study documented a rare clinical S. haemolyticus strain with three independent mechanisms of linezolid-resistance. The strain carried cfr gene, the only known transmissible mechanism of linezolid-resistance. The strain also possessed resistance-conferring mutations such as G 2576 T in domain V of 23S rRNA gene and Met 156 Thr in L3 ribosomal protein. The other ribosomal proteins (L4 and L22 did not exhibit mutations accountable for linezolid-resistance. Restriction digestion by NheI revealed that all the alleles of 23S rRNA gene were mutated. The isolate showed elevated MIC values (>256 ΅g ml -[1] of linezolid, clindamycin, chloramphenicol and oxacillin. Methicillin resistance was conferred by type I SCCmec element. The strain also harboured lsa(B gene which encodes an ABC transporter that can efflux clindamycin. Interpretation & conclusions: The present study reports the first clinical strain from India with transmissible and multiple mechanisms of linezolid-resistance. Judicious use of

  2. Triclosan-resistant bacteria isolated from feedlot and residential soils.

    Science.gov (United States)

    Welsch, Tanner T; Gillock, Eric T

    2011-01-01

    Triclosan is an antimicrobial agent that is currently incorporated into hundreds of consumer and medical products. It can be either a bacteriostatic or bactericidal agent, depending on its formulation. It has activity against Gram-positive and Gram-negative bacteria, as well as some viruses and protists. The purpose of this study was to determine whether triclosan-resistant bacteria could be isolated from the soil. Soils from cattle feedlots and residential lawns were collected and assayed for the presence of these organisms by plating samples on growth media containing triclosan. Organisms were subsequently identified by partial 16S rRNA sequencing analysis. All the organisms isolated in this study were Gram-negative rods, with members of genus Pseudomonas being particularly well represented. This result may not be surprising because Gram-negative organisms are generally more resistant to triclosan, and since Pseudomonas bacteria are known to have numerous efflux mechanisms for dealing with harmful substances.

  3. Targeted Interactomics in Plants Through Protein Complex Isolation

    Institute of Scientific and Technical Information of China (English)

    Geert De Jaeger

    2012-01-01

    TAPtag technology is the most widely applied tool to pick up in situ protein interactions in a proteome wide setting.Our research team has developed a versatile TAP technology platform for protein complex isolation from plants.We isolated complexes for hundreds of proteins and extensively demonstrated the power of our technology for protein discovery,functional analysis of proteins and protein complexes,and the modelling of protein networks.Complexes are purified from Arabidopsis cell suspension cultures or seedlings and we are currently translating the technology towards crop plants to bring complex purification in a developmental context.Besides protein complexes,we are deriving protocol variations to isolate chromatin complexes.

  4. Drug resistance. K13-propeller mutations confer artemisinin resistance in Plasmodium falciparum clinical isolates.

    Science.gov (United States)

    Straimer, Judith; Gnädig, Nina F; Witkowski, Benoit; Amaratunga, Chanaki; Duru, Valentine; Ramadani, Arba Pramundita; Dacheux, Mélanie; Khim, Nimol; Zhang, Lei; Lam, Stephen; Gregory, Philip D; Urnov, Fyodor D; Mercereau-Puijalon, Odile; Benoit-Vical, Françoise; Fairhurst, Rick M; Ménard, Didier; Fidock, David A

    2015-01-23

    The emergence of artemisinin resistance in Southeast Asia imperils efforts to reduce the global malaria burden. We genetically modified the Plasmodium falciparum K13 locus using zinc-finger nucleases and measured ring-stage survival rates after drug exposure in vitro; these rates correlate with parasite clearance half-lives in artemisinin-treated patients. With isolates from Cambodia, where resistance first emerged, survival rates decreased from 13 to 49% to 0.3 to 2.4% after the removal of K13 mutations. Conversely, survival rates in wild-type parasites increased from ≤0.6% to 2 to 29% after the insertion of K13 mutations. These mutations conferred elevated resistance to recent Cambodian isolates compared with that of reference lines, suggesting a contemporary contribution of additional genetic factors. Our data provide a conclusive rationale for worldwide K13-propeller sequencing to identify and eliminate artemisinin-resistant parasites.

  5. Antibiotic resistance of Neisseria gonorrhoeae isolated from gonorrhoeae patients

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    Mohammad Bokaeian

    2010-06-01

    Full Text Available Background: The objective of this study was to determine antibiotic susceptibility and penicillinase production by Neisseria gonorrhoeae strains isolated from gonorrhoeae patients in Zahedan.Materials and methods: In a descriptive study during 2005-2008 years, 400 suspected patients were studied by history review, medical examination, gram staining and culture in Thayer-Martin medium. Antibiotic susceptibility and penicillinase tests of isolated strains were done by disk diffusion method and aciodometric method, respectively.Results: Cultures were positive in 77(19.2% patients. The resistanat rate against antibiotics were as follow: penicillin (79.2%, ciprofloxacin (53.2%, ceftriaxone(3.8%, spectinomycin(2.5%, cefixime(12.9%, co-trimoxazole(93.5%, tetracycline(88.3% and gentamicin(29.8%. In the meanwhile, 83.1 percent of penicillin resistant isolates produced penicillinase enzyme. Discussion: Ceftriaxone, spectinomycin and cefixime are the sole antibiotics that could be considered as selective drugs. Quinolones which were regarded as an effective group of antibiotics recently have lost their importance. Resistance against other antibiotics is rapidly developing, thus, conducting experimental tests and determination of minimum inhibitory concentration and clinical trial studies at fixed intervals can contribute to diagnosis of resistance of gonococci and rapid and successful treatment of their infections in Zahedan region.

  6. Analysis of Romanian Bacteroides isolates for antibiotic resistance levels and the corresponding antibiotic resistance genes.

    Science.gov (United States)

    Székely, Edit; Eitel, Zsuzsa; Molnár, Szabolcs; Szász, Izabella Éva; Bilca, Doina; Sóki, József

    2015-02-01

    As part of an ESCMID Study Group on Anaerobic Infections (ESGAI) project, a study was conducted to measure the antibiotic susceptibilities and corresponding gene contents of 53 Bacteroides fragilis group strains isolated in Romania. The antibiotic resistance data was comparable with the data found for other East-European countries. Here, no resistant isolate was found for imipenem, metronidazole and tigecycline. An increasing role of the cepA, cfxA and cfiA genes was observed in their corresponding antibiotic resistances. Moreover, no isolate was found that harbored the cfiA gene with a possible activating IS element. Clindamycin resistance was low, similarly to that the rate for the ermF gene. However, we did find some isolates with nimB, ermB, msrSA, linA, satG, tetX, tetM and bexA genes. This study was the first to provide antibiotic resistance data for clinical Bacteroides strains from Romania.

  7. Antimicrobial drug resistance of Salmonella isolates from meat and humans, Denmark

    DEFF Research Database (Denmark)

    Skov, Marianne Nielsine; Andersen, Jens Strodl; Aabo, Søren

    2007-01-01

    We compared 8,144 Salmonella isolates collected from meat imported to or produced in Denmark, as well as from Danish patients. Isolates from imported meat showed a higher rate of antimicrobial drug resistance, including multidrug resistance, than did isolates from domestic meat. Isolates from...... humans showed resistance rates lower than those found in imported meat but higher than in domestic meat. These findings indicate that programs for controlling resistant Salmonella spp. are a global issue...

  8. Novel Aminoglycoside Resistance Transposons and Transposon-Derived Circular Forms Detected in Carbapenem-Resistant Acinetobacter baumannii Clinical Isolates.

    Science.gov (United States)

    Karah, Nabil; Dwibedi, Chinmay Kumar; Sjöström, Karin; Edquist, Petra; Johansson, Anders; Wai, Sun Nyunt; Uhlin, Bernt Eric

    2016-01-11

    Acinetobacter baumannii has emerged as an important opportunistic pathogen equipped with a growing number of antibiotic resistance genes. Our study investigated the molecular epidemiology and antibiotic resistance features of 28 consecutive carbapenem-resistant clinical isolates of A. baumannii collected throughout Sweden in 2012 and 2013. The isolates mainly belonged to clonal complexes (CCs) with an extensive international distribution, such as CC2 (n = 16) and CC25 (n = 7). Resistance to carbapenems was related to blaOXA-23 (20 isolates), blaOXA-24/40-like (6 isolates), blaOXA-467 (1 isolate), and ISAba1-blaOXA-69 (1 isolate). Ceftazidime resistance was associated with blaPER-7 in the CC25 isolates. Two classical point mutations were responsible for resistance to quinolones in all the isolates. Isolates with high levels of resistance to aminoglycosides carried the 16S rRNA methylase armA gene. The isolates also carried a variety of genes encoding aminoglycoside-modifying enzymes. Several novel structures involved in aminoglycoside resistance were identified, including Tn6279, ΔTn6279, Ab-ST3-aadB, and different assemblies of Tn6020 and TnaphA6. Importantly, a number of circular forms related to the IS26 or ISAba125 composite transposons were detected. The frequent occurrence of these circular forms in the populations of several isolates indicates a potential role of these circular forms in the dissemination of antibiotic resistance genes.

  9. Molecular characterization of multidrug resistant hospital isolates using the antimicrobial resistance determinant microarray.

    Directory of Open Access Journals (Sweden)

    Tomasz A Leski

    Full Text Available Molecular methods that enable the detection of antimicrobial resistance determinants are critical surveillance tools that are necessary to aid in curbing the spread of antibiotic resistance. In this study, we describe the use of the Antimicrobial Resistance Determinant Microarray (ARDM that targets 239 unique genes that confer resistance to 12 classes of antimicrobial compounds, quaternary amines and streptothricin for the determination of multidrug resistance (MDR gene profiles. Fourteen reference MDR strains, which either were genome, sequenced or possessed well characterized drug resistance profiles were used to optimize detection algorithms and threshold criteria to ensure the microarray's effectiveness for unbiased characterization of antimicrobial resistance determinants in MDR strains. The subsequent testing of Acinetobacter baumannii, Escherichia coli and Klebsiella pneumoniae hospital isolates revealed the presence of several antibiotic resistance genes [e.g. belonging to TEM, SHV, OXA and CTX-M classes (and OXA and CTX-M subfamilies of β-lactamases] and their assemblages which were confirmed by PCR and DNA sequence analysis. When combined with results from the reference strains, ~25% of the ARDM content was confirmed as effective for representing allelic content from both Gram-positive and -negative species. Taken together, the ARDM identified MDR assemblages containing six to 18 unique resistance genes in each strain tested, demonstrating its utility as a powerful tool for molecular epidemiological investigations of antimicrobial resistance in clinically relevant bacterial pathogens.

  10. Immunochemical studies of 22S protein from isolated mitotic apparatus.

    Science.gov (United States)

    Bibring, T; Baxandall, J

    1969-05-01

    Evidence is presented that the "22S protein" of mitotic apparatus isolated from sea urchin eggs is not microtubule protein. An antibody preparation active against 22S protein is described, and immunochemical studies of the distribution of 22S protein in various cellular fractions and among morphological features of mitotic apparatus are reported. The protein is ubiquitous in the metaphase egg fractions that were tested but is not found in sperm flagella. It is immunologically distinct from proposed microtubule protein isolated from mitotic apparatus by the method of Sakai, and from proposed microtubule protein obtained after extraction with mild acid. It exists in nontubule material of isolated mitotic apparatus but is not detectable in microtubules.

  11. Antimicrobial drug resistance of Salmonella isolates from meat and humans, Denmark

    DEFF Research Database (Denmark)

    Skov, Marianne; Andersen, Jens Strodl; Aabo, Søren

    2007-01-01

    We compared 8,144 Salmonella isolates collected from meat imported to or produced in Denmark, as well as from Danish patients. Isolates from imported meat showed a higher rate of antimicrobial drug resistance, including multidrug resistance, than did isolates from domestic meat. Isolates from hum...

  12. Efflux pump-mediated benzalkonium chloride resistance in Listeria monocytogenes isolated from retail food.

    Science.gov (United States)

    Jiang, Xiaobing; Yu, Tao; Liang, Yu; Ji, Shengdong; Guo, Xiaowei; Ma, Jianmin; Zhou, Lijun

    2016-01-18

    In this study, efflux pump-mediated benzalkonium chloride (BC) resistance, including plasmid-encoded (Qac protein family and BcrABC) and chromosome-borne efflux pumps, was investigated in Listeria monocytogenes from retail food in China. Among the 59 L. monocytogenes strains, 13 (22.0%) strains were resistant to BC. The PCR results showed that bcrABC was harbored by 2 of 13 BC resistant strains. However, none of the qac genes were detected among the 59 strains. The bcrABC was absent in both of the plasmid cured strains, indicating that this BC resistance determinant was plasmid-encoded in the two bcrABC-positive strains. In the presence of reserpine, most of the bcrABC-negative strains had decreases in the MICs of BC, suggesting the existence of other efflux pumps and their role in BC resistance. After exposed to reserpine, the reduction in BC MICs was observed in the two cured strains, indicating that efflux pumps located on chromosome was also involved in BC resistance. Our findings suggest that food products may act as reservoirs for BC resistant isolates of L. monocytogenes and plasmid- and chromosome-encoded efflux pumps could mediate the BC resistance of L. monocytogenes, which is especially relevant to the adaption of this organism in food-related environments with frequent BC use.

  13. Characterization of recombinant fluoroquinolone-resistant pneumococcus-like isolates.

    Science.gov (United States)

    Balsalobre, Luz; Ortega, Montserrat; de la Campa, Adela G

    2013-01-01

    Fourteen fluoroquinolone-resistant streptococcal isolates with recombinant DNA topoisomerase genes, preliminarily identified as pneumococci, were further characterized using phenotypic and genotypic approaches. Phenotypic tests classified them as atypical pneumococci. Phylogenetic relationships were analyzed by using the sequences of seven housekeeping alleles from these isolates and from isolates of Streptococcus pneumoniae, Streptococcus mitis, Streptococcus oralis, and Streptococcus pseudopneumoniae. Four isolates grouped with S. pneumoniae, seven grouped with S. pseudopneumoniae, and three grouped with S. mitis. These results generally agreed with those obtained with an optochin susceptibility test and with the organization of the atp operon chromosomal region, encoding the F(o)F(1) H(+)-ATPase (the target of optochin). All seven isolates grouping with S. pseudopneumoniae share the same spr1368-atpC-atpA gene order; all four grouping with S. pneumoniae share the spr1368-IS1239-atpC-atpA order, and two out of the three grouping with S. mitis share the spr1284-atpC-atpA order. In addition, evidence for recombination within the seven housekeeping alleles of the S. pseudopneumoniae population was provided by several methods: the index of association (0.4598, P < 0.001), the pairwise homoplasy index, and the split-decomposition method. This study confirms the existence of pneumococci among the alpha-hemolytic streptococci with DNA topoisomerase genes showing a mosaic structure and reveals a close relationship between atypical pneumococci and S. pseudopneumoniae.

  14. Isolated cell behavior drives the evolution of antibiotic resistance

    Science.gov (United States)

    Artemova, Tatiana; Gerardin, Ylaine; Dudley, Carmel; Vega, Nicole M; Gore, Jeff

    2015-01-01

    Bacterial antibiotic resistance is typically quantified by the minimum inhibitory concentration (MIC), which is defined as the minimal concentration of antibiotic that inhibits bacterial growth starting from a standard cell density. However, when antibiotic resistance is mediated by degradation, the collective inactivation of antibiotic by the bacterial population can cause the measured MIC to depend strongly on the initial cell density. In cases where this inoculum effect is strong, the relationship between MIC and bacterial fitness in the antibiotic is not well defined. Here, we demonstrate that the resistance of a single, isolated cell—which we call the single-cell MIC (scMIC)—provides a superior metric for quantifying antibiotic resistance. Unlike the MIC, we find that the scMIC predicts the direction of selection and also specifies the antibiotic concentration at which selection begins to favor new mutants. Understanding the cooperative nature of bacterial growth in antibiotics is therefore essential in predicting the evolution of antibiotic resistance. PMID:26227664

  15. Gentamicin resistance among Escherichia coli strains isolated in neonatal sepsis.

    Science.gov (United States)

    Hasvold, J; Bradford, L; Nelson, C; Harrison, C; Attar, M; Stillwell, T

    2013-01-01

    Neonatal sepsis is a significant cause of morbidity and mortality among term and preterm infants. Ampicillin and gentamicin are standard empiric therapy for early onset sepsis. Four cases of neonatal sepsis secondary to Escherichia coli (E. coli) found to be gentamicin resistant occurred within a five week period in one neonatal intensive care unit (NICU). To determine whether these cases could be tied to a single vector of transmission, and to more broadly evaluate the incidence of gentamicin resistant strains of E. coli in the neonatal population at our institution compared to other centers, we reviewed the charts of the four neonates (Infants A through D) and their mothers. The E. coli isolates were sent for Pulse Field Gel Electrophoresis (PFGE) to evaluate for genetic similarity between strains. We also reviewed all positive E. coli cultures from one NICU over a two year period. Infants A and B had genetically indistinguishable strains which matched that of urine and placental cultures of Infant B's mother. Infant C had a genetically distinct organism. Infant D, the identical twin of Infant C, did not have typing performed. Review of all cultures positive for E. coli at our institution showed a 12.9 percent incidence of gentamicin-resistance. A review of other studies showed that rates of resistance vary considerably by institution. We conclude that gentamicin-resistant E. coli is a relatively uncommon cause of neonatal sepsis, but should remain a consideration in patients who deteriorate despite initiation of empiric antibiotics.

  16. Overexpression of ubiquitin and amino acid permease genes in association with antimony resistance in Leishmania tropica field isolates.

    Science.gov (United States)

    Kazemi-Rad, Elham; Mohebali, Mehdi; Khadem-Erfan, Mohammad Bagher; Hajjaran, Homa; Hadighi, Ramtin; Khamesipour, Ali; Rezaie, Sassan; Saffari, Mojtaba; Raoofian, Reza; Heidari, Mansour

    2013-08-01

    The mainstay therapy against leishmaniasis is still pentavalent antimonial drugs; however, the rate of antimony resistance is increasing in endemic regions such as Iran. Understanding the molecular basis of resistance to antimonials could be helpful to improve treatment strategies. This study aimed to recognize genes involved in antimony resistance of Leishmania tropica field isolates. Sensitive and resistant L. tropica parasites were isolated from anthroponotic cutaneous leishmaniasis patients and drug susceptibility of parasites to meglumine antimoniate (Glucantime®) was confirmed using in vitro assay. Then, complementary DNA-amplified fragment length polymorphism (cDNA-AFLP) and real-time reverse transcriptase-PCR (RT-PCR) approaches were utilized on mRNAs from resistant and sensitive L. tropica isolates. We identified 2 known genes, ubiquitin implicated in protein degradation and amino acid permease (AAP3) involved in arginine uptake. Also, we identified 1 gene encoding hypothetical protein. Real-time RT-PCR revealed a significant upregulation of ubiquitin (2.54-fold), and AAP3 (2.86-fold) (P<0.05) in a resistant isolate compared to a sensitive one. Our results suggest that overexpression of ubiquitin and AAP3 could potentially implicated in natural antimony resistance.

  17. Heavy metal and disinfectant resistance genes among livestock-associated methicillin-resistant Staphylococcus aureus isolates.

    Science.gov (United States)

    Argudín, M Angeles; Lauzat, Birgit; Kraushaar, Britta; Alba, Patricia; Agerso, Yvonne; Cavaco, Lina; Butaye, Patrick; Porrero, M Concepción; Battisti, Antonio; Tenhagen, Bernd-Alois; Fetsch, Alexandra; Guerra, Beatriz

    2016-08-15

    Livestock associated methicillin-resistant Staphylococcus aureus (LA-MRSA) has emerged in animal production worldwide. Most LA-MRSA in Europe belong to the clonal complex (CC) 398. The reason for the LA-MRSA emergence is not fully understood. Besides antimicrobial agents used for therapy, other substances with antimicrobial activity applied in animal feed, including metal-containing compounds might contribute to their selection. Some of these genes have been found in various novel SCCmec cassettes. The aim of this study was to assess the occurrence of metal-resistance genes among a LA-S. aureus collection [n=554, including 542 MRSA and 12 methicillin-susceptible S. aureus (MSSA)] isolated from livestock and food thereof. Most LA-MRSA isolates (76%) carried at least one metal-resistance gene. Among the LA-MRSA CC398 isolates (n=456), 4.8%, 0.2%, 24.3% and 71.5% were positive for arsA (arsenic compounds), cadD (cadmium), copB (copper) and czrC (zinc/cadmium) resistance genes, respectively. In contrast, among the LA-MRSA non-CC398 isolates (n=86), 1.2%, 18.6% and 16.3% were positive for the cadD, copB and czrC genes, respectively, and none were positive for arsA. Of the LA-MRSA CC398 isolates, 72% carried one metal-resistance gene, and the remaining harboured two or more in different combinations. Differences between LA-MRSA CC398 and non-CC398 were statistically significant for arsA and czrC. The czrC gene was almost exclusively found (98%) in the presence of SCCmec V in both CC398 and non-CC398 LA-MRSA isolates from different sources. Regarding the LA-MSSA isolates (n=12), some (n=4) were also positive for metal-resistance genes. This study shows that genes potentially conferring metal-resistance are frequently present in LA-MRSA.

  18. Diversity of polymyxin resistance mechanisms among Acinetobacter baumannii clinical isolates.

    Science.gov (United States)

    Girardello, Raquel; Visconde, Marina; Cayô, Rodrigo; Figueiredo, Regina Célia Bressan Queiroz de; Mori, Marcelo Alves da Silva; Lincopan, Nilton; Gales, Ana Cristina

    2017-01-01

    Polymyxins have become drugs of last resort for treatment of multi-drug resistant (MDR) Gram-negative infections. However, the mechanisms of resistance to this compound have not been completely elucidated. In this study, we evaluated the mechanisms of resistance to this antimicrobial in two A. baumannii clinical isolates, respectively, susceptible (A027) and resistant (A009) to polymyxin B before and after polymyxin B exposure (A027(ind) and A009(ind)). The pmrAB and lpxACD were sequenced and their transcriptional levels were analyzed by qRT-PCR. The bacterial cell morphology was evaluated by transmission electronic microscopy (TEM) and the membrane potential was measured using Zeta-potential analyzer. The virulence of strains was studied using a Caenorhabditis elegans model. Both clinical isolates exhibited an elevation of the polymyxin B MIC after exposure to this compound. On the other hand, A027(ind) showed decreased values of MIC for β-lactams, aminoglycosides, vancomycin, teicoplanin, oxacillin and erythromycin. A027(ind) harbored two mutations in pmrB and the ISAba125 disrupting the lpxA. In contrast, A009(ind) strain exhibited increase of pmrB transcriptional level, after polymyxin B exposure, despite the absence of mutations in the pmrAB genes. The TEM images revealed a thicker and more electron-dense peptidoglycan layer for A009 than that of A027. The exposure to polymyxin B induced a strong condensation and darkening of intracellular material, mainly in A009(ind). In addition, the surface charge of A009 was significantly less negative than the one of A027. Using the C. elegans model, only A027(ind) strain showed a reduction on virulence. The diversity of polymyxin B resistance mechanisms among A. baumannii strains evaluated in this study confirms the complexity of these mechanisms, which may vary depending of the background of each strain.

  19. Antimicrobial resistance of bacterial strains isolated from avian cellulitis

    Directory of Open Access Journals (Sweden)

    MM Santos

    2014-03-01

    Full Text Available Avian cellulitis is an inflammatory process in the subcutaneous tissue, mainly located in the abdomen and thighs. This problem is commonly observed in poultry at slaughter and it is considered one of the major causes of condemnation of carcasses in Brazil. The aim of this study was to perform the microbial isolation of lesions of avian cellulitis from a processing plant located in the State of Goiás in order to analyze antimicrobial resistance by antibiogram test and to detect resistance genes by polymerase chain reaction. A total of 25 samples of avian cellulitis lesions were analyzed, from which 30 bacterial strains were isolated. There were eleven (44% strains of Escherichia coli, nine (36% strains of Staphylococcus epidermidis, seven (28% strains of Proteus mirabilis and three (12% strains of Manheimiahaemolytica. The antibiogram test showed that all strains were resistant to at least one antimicrobial. The gene of antimicrobial resistance tetB was detected in E. coli, S. epidermidis and P. mirabilis strains, and was the most frequently observed gene. The gene of antimicrobial resistance Sul1 was detected in all bacterial species, while tetA was found in E. coli and S. epidermidis strains, SHV in E. coli strains, S. epidermidis and P. mirabilis,and cat1 in one P. mirabilis strain. The results suggest a potential public health hazard due to the ability of these microorganisms to transmit antimicrobial resistancegenes to other microorganisms present in the intestinal tract of humans and animals, which may affect clinical-medical usage of these drugs.

  20. Identification of lead- resistant endophytic bacteria isolated from rice.

    Directory of Open Access Journals (Sweden)

    Alexander Pérez-Cordero

    2015-06-01

    Full Text Available   The objective of this study was to evaluate in vitro the endophytic bacteria resistance to different lead concentrations. The sampling was undertaken in the first half of 2013, when tissue samples of commercial varieties of rice at tillering stage were collected in Montería, Cordoba, Colombia. Each tissue was subjected to surface cleaning. Endophytic bacteria in agar R2A medium were isolated. Population density (CFU/g tissue was determined from each tissue, by direct counting of R2A medium surface. morphotypes were classified by shape, color, size, and appearance. A total of 168 morphotypes were isolated from root, tillers, and leaf of different commercial varieties of rice. The lead resistance test was performed in vitro, to do that, suspensions of endophytic bacteria in log phase were prepared and inoculated in minimal medium with five concentrations of lead as Pb(NO32. The experiment was incubated at 32 °C and agitated at 150 rpm, for five days. Every hour afterstarting the test, turbidimetry measuring at 600 nm was conducted. Results showed the ability of endophytic bacteria to grow at concentrations of 100% of Pb as Pb(NO32. The results of the identification with kit API20E confirmed the presence of Burkholderia cepacia and Pseudomonas putida, which showed resistance to different lead concentrations.

  1. PROFILE OF RESISTANCE OF Escherichia coli ISOLATED FROM CANINE PYOMETRA

    Directory of Open Access Journals (Sweden)

    Fernanda Santana Oliveira

    2016-10-01

    Full Text Available The endothelial pyometra is a disease that affects more frequently reproductively active adult females. Characterized by inflammation and accumulation of exudate in the uterine cavity, generally associated with bacterial infections. The present study aimed to evaluate the resistance profile of Escherichia coli isolates from 42 female dogs diagnosed with pyometra, seen at the Department of Small Animal Surgery, Hospital of Veterinary Medicine, Federal University of Bahia. To perform the bacteriological analysis, a sample of the contents of the uterus was obtained immediately after surgery of ovariosalpingohisterectomy therapy (OSH and sent to the laboratory. Microbiological analysis showed a predominance of the bacterium Escherichia coli in 40.5% (15/37. Strains of Escherichia coli isolates showed higher rates of resistance to antimicrobial erythromycin (93.3 %, azithromycin (80 %, ampicillin, amoxicillin, and cephalothin (40% each. This study reinforces the need to perform the microbiological examination for epidemiological purposes and the correct therapeutic application, thereby avoiding the indiscriminate use of antimicrobials and the potential emergence of multidrug-resistant  strains. Keywords: bacteria; multiresistant;  uterus.

  2. Genetic Lineages and Antimicrobial Resistance in Pseudomonas spp. Isolates Recovered from Food Samples.

    Science.gov (United States)

    Estepa, Vanesa; Rojo-Bezares, Beatriz; Torres, Carmen; Sáenz, Yolanda

    2015-06-01

    Raw food is a reservoir of Pseudomonas isolates that could be disseminated to consumers. The presence of Pseudomonas spp. was studied in food samples, and the phenotypic and genotypic characterizations of the recovered isolates were analyzed. Two samples of meat (3%, turkey and beef) and 13 of vegetables (22%, 7 green peppers and 6 tomatoes) contained Pseudomonas spp. A total of 20 isolates were identified, and were classified as follows (number of isolates): P. aeruginosa (5), P. putida (5), P. nitroreducens (4), P. fulva (2), P. mosselli (1), P. mendocina (1), P. monteilii (1), and Pseudomonas sp. (1). These 20 Pseudomonas isolates were clonally different by pulsed-field-gel-electrophoresis, and were resistant to the following antibiotics: ticarcillin (85%), aztreonam (30%), cefepime (10%), imipenem (10%), and meropenem (5%), but were susceptible to ceftazidime, piperacillin, piperacillin-tazobactam, doripenem, gentamicin, tobramycin, amikacin, ciprofloxacin, norfloxacin, and colistin. Only one strain (Ps158) presented a class 1 integron lacking the 3' conserved segment. The five P. aeruginosa strains were typed by multilocus sequence typing in five different sequence-types (ST17, ST270, ST800, ST1455, and ST1456), and different mutations were detected in protein OprD that were classified in three groups. One strain (Ps159) showed a new insertion sequence (ISPa47) truncating the oprD gene, and conferring resistance to imipenem.

  3. Resistance to fluoroquinolones and methicillin in ophthalmic isolates of Staphylococcus pseudintermedius from companion animals

    Science.gov (United States)

    Kang, Min-Hee; Chae, Min-Joo; Yoon, Jang-Won; Lee, So-Young; Yoo, Jong-Hyun; Park, Hee-Myung

    2014-01-01

    Resistance to fluoroquinolones and methicillin was determined for 49 ophthalmic isolates of Staphylococcus pseudintermedius from dogs with and without ophthalmic disease. Resistance was observed for ciprofloxacin (40.8%), ofloxacin (38.8%), enrofloxacin (38.8%), levofloxacin (34.7%), and moxifloxacin (4.1%). Eighteen isolates, 16 of which were resistant to oxacillin, were mecA-positive. Nine of the 16 oxacillin-resistant mecA-positive S. pseudintermedius isolates were resistant to more than one fluoroquinolone and 2 isolates were resistant to 5 fluoroquinolones. The frequency of mecA gene occurrence and fluoroquinolone resistance was twice as high among S. pseudintermedius isolates derived from dogs with ophthalmic disease compared with isolates for dogs without ophthalmic disease. The high prevalence of methicillin and fluoroquinolone resistance in S. pseudintermedius from dogs with ophthalmic disease is a concern. PMID:24982521

  4. Resistance to fluoroquinolones and methicillin in ophthalmic isolates of Staphylococcus pseudintermedius from companion animals.

    Science.gov (United States)

    Kang, Min-Hee; Chae, Min-Joo; Yoon, Jang-Won; Lee, So-Young; Yoo, Jong-Hyun; Park, Hee-Myung

    2014-07-01

    Resistance to fluoroquinolones and methicillin was determined for 49 ophthalmic isolates of Staphylococcus pseudintermedius from dogs with and without ophthalmic disease. Resistance was observed for ciprofloxacin (40.8%), ofloxacin (38.8%), enrofloxacin (38.8%), levofloxacin (34.7%), and moxifloxacin (4.1%). Eighteen isolates, 16 of which were resistant to oxacillin, were mecA-positive. Nine of the 16 oxacillin-resistant mecA-positive S. pseudintermedius isolates were resistant to more than one fluoroquinolone and 2 isolates were resistant to 5 fluoroquinolones. The frequency of mecA gene occurrence and fluoroquinolone resistance was twice as high among S. pseudintermedius isolates derived from dogs with ophthalmic disease compared with isolates for dogs without ophthalmic disease. The high prevalence of methicillin and fluoroquinolone resistance in S. pseudintermedius from dogs with ophthalmic disease is a concern.

  5. [Environmental fitness of metalaxyl-resistant isolate of Phytophthora capsici].

    Science.gov (United States)

    Wang, Guangfei; Ma, Yan

    2015-05-04

    The environmental fitness of metalaxyl-resistant isolate of Phytophthora capsici was studied for assessing the risk of metalaxyl-resistant P. capsici. We studied the main biological characteristics, competitive ability on plate, pathogenicity on pepper plant and adaptability in soil of the laboratory-induced metalaxyl-resistant isolate of P. capsici (Pc2-3 strain), with the metalaxyl-sensitive isolate (Pc2 strain, the wild-type) as the control. The zoosporangia production, releasing rate of zoosporangia and germination rate of zoospores of Pc2-3 were less than that of Pc2. The temperature range, optimum temperature range and initial pH range for mycelia growth of Pc2-3 were consistent with that of Pc2, but mycelia growth rate of Pc2-3 was lower than that of Pc2. Pc2-3 exhibited significantly weak competitive ability compared with Pc2 on carrots plate. Disease incidence of pepper inoculated with Pc2-3 (14.3%) was significantly lower than that of Pc2 (88. 6% ). When pepper plant was inoculated by mixtures of zoospore suspension of Pc2-3 and Pc2 at same ratio, the disease incidence, closing to that by Pc2 strain, was 75.7% . And all the strains isolated from diseased plants in the treatment were metalaxyl-sensitive. The density of P. capsis Pc2-3 was 0.28 times of Pc2 after the soil inoculated with Pc2-3 and Pc2 respectively at same zoospores density was incubated for 20 days. Otherwise, the ratio of Pc2-3 to Pc2 was 0.42 if the metalaxyl concentration in the soil was 300 mg/kg dry soil. No matter the soil temperature and humidity were beneficial to survival of P. capsici or not, Pc2-3 showed lower soil adaptability than Pc2. The environmental fitness of metalaxyl-resistant P. capsis Pc2-3 was weaker than the metalaxyl- sensitive strain Pc2 (the wild-type).

  6. [Drug resistance of Escherichia coli strains isolated from poultry].

    Science.gov (United States)

    Giurov, B; Korudzhiĭski, N; Bineva, I

    1981-01-01

    Studied was the sensitivity of a total of 143 strains of Escherichia coli, isolated from young birds and broilers died from coli septicaemia, to antibiotics and chemotherapeutics. The following descending order was established: gentamycin, carbenicillin, ampicillin, furazolidon, borgal, kanamycin, strep tomycin, chloramphenicol, neomycin sulphathiazole, and tetracycline. Markers of resistance were established with all strains with regard to the therapeutic agents in current and prospective use in industrial poultry farming. It is stated that a preliminary antibiogram is indispensable in order to obtain dependable results in the treatment of animals affected with colibacteriosis. An alternative is to apply directly those drugs to which the strains have shown highest sensitivity.

  7. Chloramphenicol resistant Salmonella species isolated between 1978 and 1987.

    Directory of Open Access Journals (Sweden)

    Talawadekar N

    1989-04-01

    Full Text Available During the ten year study i.e. 1978 to 1987, out of 53,251 cases of suspected enteric fever, S.typhi was isolated from 4005 cases (7% and S.para typhi ′A′ from 1108 cases (2.5%. A total of 52 strains of S. typhi and 4 strains of S.para typhi ′A′ were found to be resistant to chloramphenicol. Minimal inhibitory concentration was greater than 256 micrograms/ml of all the strains of S.typhi and S.para typhi ′A′.

  8. Molecular analysis of antimicrobial resistance mechanisms in Neisseria gonorrhoeae isolates from Ontario, Canada.

    Science.gov (United States)

    Allen, Vanessa G; Farrell, David J; Rebbapragada, Anuradha; Tan, Jingyuan; Tijet, Nathalie; Perusini, Stephen J; Towns, Lynn; Lo, Stephen; Low, Donald E; Melano, Roberto G

    2011-02-01

    Surveillance of gonococcal antimicrobial resistance and the molecular characterization of the mechanisms underlying these resistance phenotypes are essential in order to establish correct empirical therapies, as well as to describe the emergence of new mechanisms in local bacterial populations. To address these goals, 149 isolates were collected over a 1-month period (October-November 2008) at the Ontario Public Health Laboratory, Toronto, Canada, and susceptibility profiles (8 antibiotics) were examined. Mutations in previously identified targets or the presence of some enzymes related to resistance (r), nonsusceptibility (ns) (resistant plus intermediate categories), or reduced susceptibility (rs) to the antibiotics tested were also studied. A significant proportion of nonsusceptibility to penicillin (PEN) (89.2%), tetracycline (TET) (72.3%), ciprofloxacin (CIP) (29%), and macrolides (erythromycin [ERY] and azithromycin; 22.3%) was found in these strains. Multidrug resistance was observed in 18.8% of the collection. Although all the strains were susceptible to spectinomycin and extended-spectrum cephalosporins (ESC) (ceftriaxone and cefixime), 9.4% of them displayed reduced susceptibility to extended-spectrum cephalosporins. PBP 2 mosaic structures were found in all of these ESC(rs) isolates. Alterations in the mtrR promoter, MtrR repressor (TET(r), PEN(ns), ESC(rs), and ERY(ns)), porin PIB (TET(r) and PEN(ns)), and ribosomal protein S10 (TET(r)) and double mutations in gyrA and parC quinolone resistance-determining regions (QRDRs) (CIP(r)) were associated with and presumably responsible for the resistance phenotypes observed. This is the first description of ESC(rs) in Canada. The detection of this phenotype indicates a change in the epidemiology of this resistance and highlights the importance of continued surveillance to preserve the last antimicrobial options available.

  9. Protein profile of Chlamydophila abortus isolates from Kerala, India

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    Binu K Mani

    Full Text Available Chlamydiae are of microbiological interest because of their mode of interaction with eukaryotic host cells and their specialized life cycle with unique features of parasitism. Reports regarding prevalence of infections of Chlamydophila abortus, the causative organism for chlamydial abortions in livestock, was the basis of the study. Two isolates, one each from cattle and goat abortion along with a reference isolate, were used for characterization with Sodium Dodecyl Sulphate-Poly Acrylamide Gel Electrophoresis (SDS-PAGE. Elementary bodies infected Mc Coy cells, harvested from bottle cultures were disrupted by Teflon coated magnetic pellet. Urografin-76 diluted with Tris-Potassium hydrochloride was used for purification of Elementary bodies of Chlamydophila abortus organism. On protein estimation of Elementary bodies by Biuret method, all the three isolates revealed protein concentration between 500-1000 mg/100ml, which were sufficient for electrophoresis. Ten percent of resolving gel and five percent of stacking gel of polyacrylamide in which 10g of processed isolate samples along with standard protein marker and Mc Coy cell protein (control were electrophoresed. Using Alpha Imager Gel Documentation System, the protein bands were analyzed. Twelve bands each for local bovine isolate and reference isolate were noticed while only 10 bands were there in the caprine isolate. Additional bands of 148 kDa and 135 kDa were present in bovine isolate, compared to the reference isolate, while 152 kDa and 137 kDa bands were unique for caprine isolate. [Vet. World 2011; 4(10.000: 470-472

  10. Isolation and Determination of Antibiotic Resistance Patterns in Nontyphoid Salmonella spp isolated from chicken

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    Seyyedeh Hoorieh Fallah

    2013-01-01

    Full Text Available Background: Salmonellosis is one of the most common food borne diseases in industrial and developing countries. In recent years, an increase in antimicrobial drug resistance, among non-typhoid Salmonella spp has been observed. Objectives: The aim of this study was to isolate and determine antibiotic resistance pattern in non-typhoid Salmonella spp. Materials and Methods: This descriptive study was done on 100 samples of chickens collected from 196 retail markets and was examined for the presence of Salmonella using standard bacteriological procedures and stereotyping kit. Antimicrobial susceptibility testing was performed by disk diffusion methods according to the National Committee for Clinical Laboratory Standards (CLSI. The data were analyzed by using the SPSS software version 18. Result: Forty- four percent of samples were contaminated with Salmonella infection and 56% didn’t have any contamination. The stereotyping results showed that 34 of 44 isolates of Salmonella belonged to Salmonella infantis (79.5 %, one strain (2.3% of group C and 8 strain (18.2% of group D. However, all these strains were sensitive to Cefotaxime and Ciprofloxacin, and 100% were resistant to Nalidixic acid, Tetracyclin and Sterptomycin. The most common resistance pattern (34.1% was towards six antibiotics, and 6.8% of strains were resistant to at least three antibiotics. Conclusion: High levels of resistance to antibiotics that are used commonly for human and poultry can be a warning for our community health and this information must be used to form important strategies for improvement of infection control.

  11. Variation in the Coat Protein Gene of Papaya ringspot Virus Isolates from Multiple Locations of China

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The potyvirus Papaya ringspot virus (PRSV) is an important pathogen of papaya that causes severe losses in economic crops for papaya production globally. The coat protein (CP) genes of five PRSV isolates originating from different locations in China were cloned and sequenced. The CP-coding region varied in size from 864-873 nucleotides, encoding proteins of 288-291 amino acids. The five Chinese isolates of PRSV have been characterized as papaya-infecting (PRSV-P). The CP sequences of the Chinese isolates were compared with those of previously published PRSV isolates originating from different countries at amino acid levels. A number of KE repeat boxes in the N terminus of the PRSV-CP were found in all Chinese isolates. The phylogenetic branching pattern revealed that there was certain extended grouping between geographic locations, and the Asian type probably represents the oldest population of PRSV. The information of CP genes will be useful in designing and developing durable virus resistant-PRSV transgenic papaya in China. Meanwhile broad-spectrum-virus resistant, strongly resistant-PRSV and good safe papaya lines are required.

  12. Coagulase-negative staphylococci (CoNS) isolated from ready-to-eat food of animal origin--phenotypic and genotypic antibiotic resistance.

    Science.gov (United States)

    Chajęcka-Wierzchowska, Wioleta; Zadernowska, Anna; Nalepa, Beata; Sierpińska, Magda; Łaniewska-Trokenheim, Łucja

    2015-04-01

    The aim of this work was to study the pheno- and genotypical antimicrobial resistance profile of coagulase negative staphylococci (CoNS) isolated from 146 ready-to-eat food of animal origin (cheeses, cured meats, sausages, smoked fishes). 58 strains were isolated, they were classified as Staphylococcus xylosus (n = 29), Staphylococcus epidermidis (n = 16); Staphylococcus lentus (n = 7); Staphylococcus saprophyticus (n = 4); Staphylococcus hyicus (n = 1) and Staphylococcus simulans (n = 1) by phenotypic and genotypic methods. Isolates were tested for resistance to erythromycin, clindamycin, gentamicin, cefoxitin, norfloxacin, ciprofloxacin, tetracycline, tigecycline, rifampicin, nitrofurantoin, linezolid, trimetoprim, sulphamethoxazole/trimethoprim, chloramphenicol, quinupristin/dalfopristin by the disk diffusion method. PCR was used for the detection of antibiotic resistance genes encoding: methicillin resistance--mecA; macrolide resistance--erm(A), erm(B), erm(C), mrs(A/B); efflux proteins tet(K) and tet(L) and ribosomal protection proteins tet(M). For all the tet(M)-positive isolates the presence of conjugative transposons of the Tn916-Tn1545 family was determined. Most of the isolates were resistant to cefoxitin (41.3%) followed by clindamycin (36.2%), tigecycline (24.1%), rifampicin (17.2%) and erythromycin (13.8%). 32.2% staphylococcal isolates were multidrug resistant (MDR). All methicillin resistant staphylococci harboured mecA gene. Isolates, phenotypic resistant to tetracycline, harboured at least one tetracycline resistance determinant on which tet(M) was most frequent. All of the isolates positive for tet(M) genes were positive for the Tn916-Tn1545 -like integrase family gene. In the erythromycin-resistant isolates, the macrolide resistance genes erm(C) or msr(A/B) were present. Although coagulase-negative staphylococci are not classical food poisoning bacteria, its presence in food could be of public health significance due to the possible spread of

  13. Resistance breaking tomato spotted wilt virus isolates on resistant pepper varieties in Italy.

    Science.gov (United States)

    Crescenzi, A; Viggiano, A; Fanigliulo, A

    2013-01-01

    In spring 2012, resistance breaking (RB) isolates of tomato spotted wilt virus (TSWV) that overcome the resistance conferred by the Tsw gene in different pepper hybrids have been recovered in different locations in southern Italy (Campania and Apulia regions) in protected cultivation, about one month after transplant. The percentage of symptomatic plants was 5-10% and, only in particular cases of advanced stage of cultivation, it reached 30-50% at the end of cycle. All TSWV isolates induced similar systemic symptoms in all resistant infected pepper hybrids: yellowing or browning of apical leaves, which later become necrotic, long necrotic streakson stems, extending to the terminal shoots, complete necrosis of younger fruits and large necrotic streaks and spots on fruits formed after infection. On ripe fruits, yellow spots with concentric rings or necrotic streaks could be observed. Leaf extracts of these samples were tested in ELISA for the detection of TSWV, Cucumber mosaic virus (CMV), Tobacco mosaic virus (TMV), Tomato mosaic virus (ToMV), Impatiens necrotic spot virus (INSV), Potato virus Y (PVY), Alfalfa mosaic virus (AMV), Pepper mild mottle virus (PMMoV) and Pepper Mottle Virus (PepMoV). Only TSWV was detected in all the field samples tested. The correspondent virus isolates were inoculated mechanically and by Frankliniella occidentalis on to a set of different pepper and tomato hybrids, as well as on some herbaceous test plants, in order to investigate for their ability to overcome the resistance genes Tsw and Sw5, respectively. Tomato hybrids carrying the Sw5 gene were uninfected by all RB isolates, whereas all resistant pepper hybrids became systemically infected. RB isolates did not differ noticeably in transmission efficiency when they were tested with the thrips F. occidentalis. Obtained results demonstrate that evolved strains of TSWV have emerged, that they are able to overcome the Tsw resistance gene in pepper plants experimentally inoculated both

  14. Emergence of fluoroquinolones-resistant strains of Salmonella typhi: Watch on multidrug-resistant isolates

    Directory of Open Access Journals (Sweden)

    Subhash C Arya

    2010-05-01

    Full Text Available Subhash C Arya, Nirmala Agarwal, Shekhar Agarwal, Dolly WadhwaSant Parmanand Hospital, Delhi, IndiaEmergence of multidrug-resistant Salmonella typhi has been responsible for clinical challenges for clinicians. Recently, frequent isolation and dissemination of fluoroquinolones-resistant strains of S. enterica in Surabaya, Indonesia was in the news. Subsequently, Yangai and colleagues1 recommended regular communications between laboratory professionals and clinicians. Collaboration between laboratory personnel and clinicians would be essential to offer a rational empiric antibiotic recipe while awaiting antibiotic susceptibility test results (AST for any patient.

  15. Rheological properties of soybean protein isolate gels containing emulsion droplets

    NARCIS (Netherlands)

    Kim, K.H.; Renkema, J.M.S.; Vliet, van T.

    2001-01-01

    Rheological properties of soybean protein gels containing various volume fractions oil droplets have been studied at small and large deformations. Dynamic viscoelastic properties of soybean protein isolate gels were determined as a function of the volume fraction of oil droplets stabilised by the sa

  16. Comparative Proteomic Analysis of Aminoglycosides Resistant and Susceptible Mycobacterium tuberculosis Clinical Isolates for Exploring Potential Drug Targets.

    Science.gov (United States)

    Sharma, Divakar; Kumar, Bhavnesh; Lata, Manju; Joshi, Beenu; Venkatesan, Krishnamurthy; Shukla, Sangeeta; Bisht, Deepa

    2015-01-01

    Aminoglycosides, amikacin (AK) and kanamycin (KM) are second line anti-tuberculosis drugs used to treat tuberculosis (TB) and resistance to them affects the treatment. Membrane and membrane associated proteins have an anticipated role in biological processes and pathogenesis and are potential targets for the development of new diagnostics/vaccine/therapeutics. In this study we compared membrane and membrane associated proteins of AK and KM resistant and susceptible Mycobacterium tuberculosis isolates by 2DE coupled with MALDI-TOF/TOF-MS and bioinformatic tools. Twelve proteins were found to have increased intensities (PDQuest Advanced Software) in resistant isolates and were identified as ATP synthase subunit alpha (Rv1308), Trigger factor (Rv2462c), Dihydrolipoyl dehydrogenase (Rv0462), Elongation factor Tu (Rv0685), Transcriptional regulator MoxR1(Rv1479), Universal stress protein (Rv2005c), 35kDa hypothetical protein (Rv2744c), Proteasome subunit alpha (Rv2109c), Putative short-chain type dehydrogenase/reductase (Rv0148), Bacterioferritin (Rv1876), Ferritin (Rv3841) and Alpha-crystallin/HspX (Rv2031c). Among these Rv2005c, Rv2744c and Rv0148 are proteins with unknown functions. Docking showed that both drugs bind to the conserved domain (Usp, PspA and SDR domain) of these hypothetical proteins and GPS-PUP predicted potential pupylation sites within them. Increased intensities of these proteins and proteasome subunit alpha might not only be neutralized/modulated the drug molecules but also involved in protein turnover to overcome the AK and KM resistance. Besides that Rv1876, Rv3841 and Rv0685 were found to be associated with iron regulation signifying the role of iron in resistance. Further research is needed to explore how these potential protein targets contribute to resistance of AK and KM.

  17. Novel mechanism of bacteriocin secretion and immunity carried out by lactococcal multidrug resistance proteins

    NARCIS (Netherlands)

    Gajic, O; Buist, G; Kojic, M; Topisirovic, L; Kuipers, OP; Kok, J

    2003-01-01

    A natural isolate of Lactococcus lactis was shown to produce two narrow spectrum class II bacteriocins, designated LsbA and LsbB. The cognate genes are located on a 5.6-kb plasmid within a gene cluster specifying LmrB, an ATP-binding cassette-type multidrug resistance transporter protein. LsbA is a

  18. Novel mechanism of bacteriocin secretion and immunity carried out by lactococcal multidrug resistance proteins

    NARCIS (Netherlands)

    Gajic, O; Buist, G; Kojic, M; Topisirovic, L; Kuipers, OP; Kok, J

    2003-01-01

    A natural isolate of Lactococcus lactis was shown to produce two narrow spectrum class II bacteriocins, designated LsbA and LsbB. The cognate genes are located on a 5.6-kb plasmid within a gene cluster specifying LmrB, an ATP-binding cassette-type multidrug resistance transporter protein. LsbA is a

  19. Multiple Antimicrobial Resistance and Novel Point Mutation in Fluoroquinolone-Resistant Escherichia coli Isolates from Mangalore, India.

    Science.gov (United States)

    Kogaluru Shivakumaraswamy, Santhosh; Vijaya Kumar, Deekshit; Moleyuru Nagarajappa, Venugopal; Karunasagar, Iddya; Karunasagar, Indrani

    2017-04-26

    Fluoroquinolone resistance in bacteria is usually associated with mutations in the topoisomerase regions. We report a novel point mutation in fluoroquinolone-resistant Escherichia coli strains. E. coli isolated from the environment in and around Mangalore, India, were examined for their antimicrobial resistance profile to 12 antibiotics and for the antibiotic resistance genes by polymerase chain reaction. Of the 67 E. coli isolated, 24 (35.8%) were sensitive to all antibiotics and 43 (64.2%) showed resistance to at least one of the 12 antibiotics used in the study. One isolate (EC10) was resistant to nine of the 12 antibiotics used. Resistance to nalidixic acid was the most common (34.32%), followed by nitrofurantoin (26.86%), tetracycline (22.38%), ampicillin (20.89%), cotrimoxazole (13.43%), ciprofloxacin (11.94%), gentamicin (10.44%), piperacillin/tazobactam (7.46%), chloramphenicol (7.46%), and cefotaxime (4.47%). Least resistance was observed for meropenem (1.49%) and none of the isolates showed resistance to imipenem. All the isolates harbored resistance genes corresponding to their antimicrobial resistance. Few quinolone-resistant isolates carried single point mutation (ser83Leu) and some had double point mutation (Ser83Leu and Asp87Asn) in gyrA. A third novel point mutation was also observed at position 50 with the change in the amino acid from tyrosine to cysteine (Tyr50Cys) in gyrA region. The study throws light on a novel point mutation in fluoroquinolone-resistant isolates. While the study helps to understand the risk and occurrence of antibiotic resistance among gram-negative bacteria from the environment, the alarming rate of antibiotic-resistant bacteria is a cause of concern in addressing infections.

  20. Quantitative proteomic analysis of ofloxacin resistant and sensitive clinical isolates of Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Xiang-yu HUANG

    2014-10-01

    Full Text Available Objective To identify the proteins related to ofloxacin (OFX resistance of Mycobacterium tuberculosis (MTB. Methods Standard MTB H37Rv strain, clinical isolates of OFX resistant strain (OFXR and sensitive strain (OFXS were obtained from the Chinese Center for Disease Control and Prevention, and they were cultured in Sauton's medium, and then inactivated by 60Co. Whole cellular proteins were extracted from OFXR, OFXS and H37Rv strain of MTB, respectively. The peptides were labeled, separated and identified by isobaric tags of relative and absolute quantitation (iTRAQ combined with Nano LCMS/MS technology. Results One hundred and seventy-five and 134 differential expression proteins were identified in MTB OFXR compared with MTB OFXS and H37Rv, respectively. One hundred and four common differential expression proteins were identified in MTB OFXR compared with both MTB OFXS and H37Rv. The isoelectric point and theoretic relative molecular mass of differential expression proteins were widely distributed. The majority of the common differential expression proteins were involved in intermediary metabolism, respiration, and lipid metabolism. Twelve common differential expression proteins showed significant differences (the ratio>1.2 or <0.55 in MTB OFXR, including Rv0106, Rv0895, Rv2185c, Rv3248c and Rv3841 up-regulation and Rv2524c, Rv2986c, Rv3118 and Rv3597c down-regulation. Conclusion iTRAQ has been used to identify the common differential expression proteins in MTB OFXR compared with both MTB OFXS and H37Rv, which provides a basis for further study of the mechanism of OFX-resistance. DOI: 10.11855/j.issn.0577-7402.2014.09.06

  1. Studies of acid resistance characteristics in multiple drug resistant Salmonella species isolated from tomatoes.

    Science.gov (United States)

    Naushad, Z; Mishra, S H; Musaddiq, M; Ali, Y A

    2013-04-01

    Salmonella species found to have a great potential of causing a variety of diseases ranging from gastroenteritis to enteric fever. Salmonella have been isolated from all food, animals and also found in the vegetables such as tomatoes, spinach etc. Several out breaks of Salmonellosis have been associated with the consumption of raw tomatoes. This is because of the fact that Salmonella attaches to the surface of tomatoes and also present in the interior part due to geotropic transmission via contaminated soil irrigated with contaminated water. .During the life cycle, Salmonella encounters the various environments such as acidic environment (low pH). To overcome such factors, Salmonella has certain adaptable mechanisms. In present 'study total 200 samples of tomatoes were analyzed out of which 10 samples were found to contain Salmonella. All the 10 isolates were then subjected to the antibiotic susceptibility testing and were found to be resistant against several antibiotics. These were subjected to acid resistant tolerance study.

  2. Proteomic analysis of streptomycin resistant and sensitive clinical isolates of Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Venkatesan Krishnamurthy

    2010-11-01

    Full Text Available Abstract Background Streptomycin (SM is a broad spectrum antibiotic and is an important component of any anti-tuberculosis therapy regimen. Several mechanisms have been proposed to explain the emergence of resistance but still our knowledge is inadequate. Proteins form a very complex network and drugs are countered by their modification/efflux or over expression/modification of targets. As proteins manifest most of the biological processes, these are attractive targets for developing drugs, immunodiagnostics or therapeutics. The aim of present study was to analyze and compare the protein profile of whole cell extracts from Mycobacterium tuberculosis clinical isolates susceptible and resistant to SM. Results Two-dimensional gel electrophoresis (2DE and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF mass spectrometry was employed for analyzing the protein profiles. Homology and in silico characterization for identified proteins was assessed using BLAST, InterProScan and KEGG database searches. Computational studies on the possible interactions between SM and identified proteins were carried out by a battery of online servers and softwares, namely, CLUSTALW (KEGG, I-TASSER, VMD, PatchDock and FireDock. On comparing 2DE patterns, nine proteins were found consistently overexpressed in SM resistant isolates and were identified as Rv0350, Rv0440, Rv1240, Rv3075c, Rv2971, Rv3028c, Rv2145c, Rv2031c and Rv0569. In silico docking analysis showed significant interactions of SM with essential (Rv0350, Rv0440 and Rv2971 and non essential (Rv1240, Rv3075c and Rv2031c genes. Conclusions The computational results suggest high protein binding affinity of SM and suggested many possible interactions between identified proteins and the drug. Bioinformatic analysis proves attributive for analysis of diversity of proteins identified by whole proteome analysis. In-depth study of the these proteins will give an insight into probable sites of drug

  3. Serovars of Salmonella isolated from Danish turkeys between 1995 and 2000 and their antimicrobial resistance

    DEFF Research Database (Denmark)

    Pedersen, Karl; Hansen, H.C.; Jørgensen, J.C.

    2002-01-01

    , florfenicol, or amoxycillin with clavulanic acid, only 24 isolates were resistant to two or more compounds in various combinations of up to six compounds; one Salmonella Havana isolate was resistant to six compounds. Six isolates were serovar Typhimurium, but none of them belonged to phage type DT104....

  4. Synergistic activity of chloroquine with fluconazole against fluconazole-resistant isolates of Candida species.

    Science.gov (United States)

    Li, Yali; Wan, Zhe; Liu, Wei; Li, Ruoyu

    2015-02-01

    The in vitro activity of chloroquine and the interactions of chloroquine combined with fluconazole against 37 Candida isolates were tested using the broth microdilution, disk diffusion, and Etest susceptibility tests. Synergistic effect was detected with 6 of 9 fluconazole-resistant Candida albicans isolates, with Candida krusei ATCC 6258, and with all 12 fluconazole-resistant Candida tropicalis isolates.

  5. Antimicrobial resistance trends in blood culture positive Salmonella Paratyphi A isolates from Pondicherry, India

    NARCIS (Netherlands)

    G.A. Menezes (Godfred A.); B.N. Harish (Belgode Narasimha); M.A. Khan (M.); W.H.F. Goessens (Wil); J.P. Hays (John)

    2016-01-01

    textabstractEnteric fever is a public health problem with the upsurge in the occurrence of Salmonella isolates that are resistant to ciprofloxacin. In this study, a total of 284 blood culture isolates of S. Paratyphi A were investigated. Of these isolates, 281 (98.9%) were nalidixic acid resistant.

  6. Molecular Genetics of para-Aminosalicylic Acid Resistance in Clinical Isolates and Spontaneous Mutants of Mycobacterium tuberculosis▿ †

    Science.gov (United States)

    Mathys, Vanessa; Wintjens, René; Lefevre, Philippe; Bertout, Julie; Singhal, Amit; Kiass, Mehdi; Kurepina, Natalia; Wang, Xiao-Ming; Mathema, Barun; Baulard, Alain; Kreiswirth, Barry N.; Bifani, Pablo

    2009-01-01

    The emergence of Mycobacterium tuberculosis resistant to first-line antibiotics has renewed interest in second-line antitubercular agents. Here, we aimed to extend our understanding of the mechanisms underlying para-aminosalicylic acid (PAS) resistance by analysis of six genes of the folate metabolic pathway and biosynthesis of thymine nucleotides (thyA, dfrA, folC, folP1, folP2, and thyX) and three N-acetyltransferase genes [nhoA, aac(1), and aac(2)] among PAS-resistant clinical isolates and spontaneous mutants. Mutations in thyA were identified in only 37% of the clinical isolates and spontaneous mutants. Overall, 24 distinct mutations were identified in the thyA gene and 3 in the dfrA coding region. Based on structural bioinformatics techniques, the altered ThyA proteins were predicted to generate an unfolded or dysfunctional polypeptide. The MIC was determined by Bactec/Alert and dilution assay. Sixty-three percent of the PAS-resistant isolates had no mutations in the nine genes considered in this study, revealing that PAS resistance in M. tuberculosis involves mechanisms or targets other than those pertaining to the biosynthesis of thymine nucleotides. The alternative mechanism(s) or pathway(s) associated with PAS resistance appears to be PAS concentration dependent, in marked contrast to thyA-mutated PAS-resistant isolates. PMID:19237648

  7. Inhibition of neuraminidase inhibitor-resistant influenza virus by DAS181, a novel sialidase fusion protein.

    Directory of Open Access Journals (Sweden)

    Gallen B Triana-Baltzer

    Full Text Available Antiviral drug resistance for influenza therapies remains a concern due to the high prevalence of H1N1 2009 seasonal influenza isolates which display H274Y associated oseltamivir-resistance. Furthermore, the emergence of novel H1N1 raises the potential that additional reassortments can occur, resulting in drug resistant virus. Thus, additional antiviral approaches are urgently needed. DAS181 (Fludase, a sialidase fusion protein, has been shown to have inhibitory activity against a large number of seasonal influenza strains and a highly pathogenic avian influenza (HPAI strain (H5N1. Here, we examine the in vitro activity of DAS181 against a panel of 2009 oseltamivir-resistant seasonal H1N1 clinical isolates. The activity of DAS181 against nine 2009, two 2007, and two 2004 clinical isolates of seasonal IFV H1N1 was examined using plaque number reduction assay on MDCK cells. DAS181 strongly inhibited all tested isolates. EC50 values remained constant against isolates from 2004, 2007, and 2009, suggesting that there was no change in DAS181 sensitivity over time. As expected, all 2007 and 2009 isolates were resistant to oseltamivir, consistent with the identification of the H274Y mutation in the NA gene of all these isolates. Interestingly, several of the 2007 and 2009 isolates also exhibited reduced sensitivity to zanamivir, and accompanying HA mutations near the sialic acid binding site were observed. DAS181 inhibits IFV that is resistant to NAIs. Thus, DAS181 may offer an alternative therapeutic option for seasonal or pandemic IFVs that become resistant to currently available antiviral drugs.

  8. Streptogramin resistance among Enterococcus faecium isolated from production animals in Denmark in 1997

    DEFF Research Database (Denmark)

    Jensen, Lars Bogø; Hammerum, Anette Marie; Bager, Flemming;

    2002-01-01

    The genetic background for streptogramin resistance was examined in Enterococcus faecium isolated from pigs (n = 55) and broilers (n = 207) in 1997 in Denmark. Fifty-one percent and 67%, respectively, of the isolates were resistant to streptogramins. Among streptogramin-resistant E. faecium (SREF...

  9. Prevalence and antibiotic resistance of Enterococcus strains isolated from poultry.

    Science.gov (United States)

    Stępień-Pyśniak, Dagmara; Marek, Agnieszka; Banach, Tomasz; Adaszek, Łukasz; Pyzik, Ewelina; Wilczyński, Jarosław; Winiarczyk, Stanisław

    2016-06-01

    The aim of this study was to evaluate the frequency of occurrence of bacteria of the genus Enterococcus in poultry, to identify them by means of matrixassisted laser desorption/ionisation time-of-flight mass spectrometry (MALDITOF MS), and to analyse the antimicrobial susceptibility of the isolated strains to the drugs most frequently used in poultry. The material for the bacteriological tests was obtained mainly from the heart (97%) of the birds investigated. Of a total of 2,970 samples tested, 911 (30.7%) tested positive for Enterococcus spp. Enterococci were detected in broilers (88.1%), laying hens (5.3%), turkeys (3.9%), breeding hens (2.2%), and geese (0.4%). The most commonly identified species were Enterococcus (E.) faecalis (74.7%), E. faecium (10.1%), E. gallinarum (5.5%), E. hirae (4.6%), and E. cecorum (4.1%). The most frequent resistance properties were resistance to sulphamethoxazole/trimethoprim (88%), tylosin (71.4%), enrofloxacin (69.4%), doxycycline (67.3%), and lincomycin/spectinomycin (56.1%). Only one vancomycin-resistant Enterococcus, E. cecorum from a broiler, was found.

  10. [The review of carbapenem resistance in clinical isolates of Acinetobacter baumannii].

    Science.gov (United States)

    Goić-Barisić, Ivana; Tonkić, Marija

    2009-10-01

    Increasing reports of Acinetobacter infections that cause pneumonia, meningitis, endocarditis, and bacteriaemia underline the clinical importance of this pathogen. Members of the genus Acinetobacter, particularly Acinetobacter baumannii, are now recognized as significant nosocomial pathogens, particularly for the subset of critically-ill patients requiring mechanical ventilation in hospital intensive care units. A. baumannii has itself a quite high level of naturally-occurring antibiotic resistance. The organism can survive for long periods in the hospital environment including dry and humid areas. One of the most worrying antibiotic resistance problems in A. baumannii is the increasing trend of carbapenem resistance, present also in few Croatian hospitals. Infections caused by this Gram-negative bacillus are common in the intensive care units anticipated by colonized patients. The increasing trend of carbapenem resistance in A. baumannii could be mediated from metallo-beta-lactamases (VIM, IMP, and SIM), carbapenem-hydrolyzing oxacillinases (OXA), porin modifications for influx of carbapenems (33-kDa CarO protein) and/or often combined mechanisms of resistance. The investigation of the background of carbapenem resistance in relevant clinical isolates of A. baumannii from Split University Hospital confirmed present of carbapenem-hydrolyzing oxacillinases OXA-107 representing a more recent evolutionary adaptation OXA-51-like enzyme to antibiotic challenge with carbapenems.

  11. Molecular epidemiology and antimicrobial resistance of methicillin-resistant Staphylococcus aureus bloodstream isolates in Taiwan, 2010.

    Directory of Open Access Journals (Sweden)

    Chih-Jung Chen

    Full Text Available The information of molecular characteristics and antimicrobial susceptibility pattern of methicillin-resistant Staphylococcus aureus (MRSA is essential for control and treatment of diseases caused by this medically important pathogen. A total of 577 clinical MRSA bloodstream isolates from six major hospitals in Taiwan were determined for molecular types, carriage of Panton-Valentine leukocidin (PVL and sasX genes and susceptibilities to 9 non-beta-lactam antimicrobial agents. A total of 17 genotypes were identified in 577 strains by pulsotyping. Five major pulsotypes, which included type A (26.2%, belonging to sequence type (ST 239, carrying type III staphylococcal chromosomal cassette mec (SCCmec, type F (18.9%, ST5-SCCmecII, type C (18.5%, ST59-SCCmecIV, type B (12.0%, ST239-SCCmecIII and type D (10.9%, ST59-SCCmecVT/IV, prevailed in each of the six sampled hospitals. PVL and sasX genes were respectively carried by ST59-type D strains and ST239 strains with high frequencies (93.7% and 99.1%, respectively but rarely detected in strains of other genotypes. Isolates of different genotypes and from different hospitals exhibited distinct antibiograms. Multi-resistance to ≥3 non-beta-lactams was more common in ST239 isolates (100% than in ST5 isolates (97.2%, P = 0.0347 and ST59 isolates (8.2%, P<0.0001. Multivariate analysis further indicated that the genotype, but not the hospital, was an independent factor associated with muti-resistance of the MRSA strains. In conclusion, five common MRSA clones with distinct antibiograms prevailed in the major hospitals in Taiwan in 2010. The antimicrobial susceptibility pattern of invasive MRSA was mainly determined by the clonal distribution.

  12. Comparison of the Resistance of Diabetic Foot Isolates with Community and Intensive Care Unit Resistance Rates

    Directory of Open Access Journals (Sweden)

    Filiz Pehlivanoğlu

    2011-12-01

    Full Text Available Objective: Using proper antibiotics in diabetic foot infections can save an extremity. The goal of this study was to determine the antibiotic susceptibility of the most frequent isolates from diabetic foot infections, to compare these results with those for species obtained from both intensive care unit and community-acquired infections, and to re-evaluate the empirical antimicrobial therapy in diabetic foot infections. Material and Methods: Antibiotic susceptibility testing was performed on bacteria from diabetic foot cultures, ICU and community-acquired infections. Skin and soft tissue samples of a total of 181 patients have been studied. Sensitivity to ampicillin/ sulbactam (SAM, ciprofloxacin (CIP, piperacillin/tazobactam (TZP, cefoperazone/sulbactam (SCF, amikacin (AN, meropenem (MEM and cefepime (FEP has been investigated. Results: Among the samples, reproduction was detected in 154 (85%, 132 of which (79.5% contained gram-negative bacteria (46 Enterobacter spp, 36 Pseudomonas spp, 30 Acinetobacter spp, 20 others. Resistance of bacteria isolated from diabetic foot cultures was less than the one of bacteria isolated from ICU and more than the one from community-acquired infections. However, an increased resistance was detected to ciprofloxacin, frequently used in urinary infections, in agents from community-acquired infections. Conclusion: In conclusion, due to the risk of possible loss of tissue/extremity and possible antibiotic resistance, treatment should be started empirically and continued considering the culture results. (The Me di cal Bul le tin of Ha se ki 2011; 49: 137-40

  13. Molecular typing and resistance mechanisms of carbapenem resistant Pseudomonas aeruginosa isolated from a Chinese surgical intensive care unit

    Institute of Scientific and Technical Information of China (English)

    Yi Meiying; Wang Pengyuan; Liu Yucun

    2014-01-01

    Background Carbapenems are an important class of drugs for the treatment of Pseudomonas aeruginosa (P.aeruginosa) infections.However,carbapenem resistance has been commonly observed in nonfermenter species of bacteria.The purpose of this study was to investigate the molecular epidemiology and carbapenem resistant mechanisms of P.aeruginosa isolated from a surgical intensive care unit (SICU) in China.Methods The molecular typing was analyzed by REP-PCR.Enzyme activity was measured with a 260 nm wavelength spectrophotometer.The levels of outer membrane proteins OprD and OprN were measured by Western blotting.The levels of mexA gene transcriptional expression were measured by quantitative real-time PCR.The metallo-beta-lactamase genes IMP,VIM,SPM,GES,and GIM were amplified by PCR.DNA fragments were sequenced by an automated ABI PRISM 3700.Results Forty-two strains resistant to carbapenems isolated from a SICU were analyzed.REP-PCR revealed 34 belonging to type A,a predominant strain in this SICU.But we did not find metallo-beta-lactamases IMP,VIM,SPM,GES,or GIM genes by PCR.With a three-dimensional extract test,we found 34 strains producing high levels of AmpC enzymes.We also observed the activity of beta-lactamases enzymes in the imipenem resistant group,which was statistically different from the sensitive group.Western blotting revealed that 23 strains showed loss of OprD,18 strains had decreased OprD expression,and 14 strains expressed OprN.We discovered 27 strains that overexpressed mexA by quantitative real-time PCR,and the resistance rate to meropenem was statistically different between the overexpressing group and the lowexpressing group.Nucleotide sequences and deduced amino acid sequence analysis revealed that eight strains carried mutations in the mexR gene operon down regulating MexAB-OprM.The nucleotide sequences of mexR genes from PA36,PA41 and PA48 were submitted to the Genebank with accession numbers of AY899299,AY899300,and AY899301.Conclusions There

  14. Characterization of antimicrobial resistance in Salmonella enterica strains isolated from Brazilian poultry production.

    Science.gov (United States)

    Mattiello, Samara P; Drescher, Guilherme; Barth, Valdir C; Ferreira, Carlos A S; Oliveira, Sílvia D

    2015-11-01

    Antimicrobial resistance profiles and presence of resistance determinants and integrons were evaluated in Salmonella enterica strains from Brazilian poultry. The analysis of 203 isolates showed that those from the poultry environment (88 isolates) were significantly more resistant to antimicrobials than isolates from other sources, particularly those isolated from poultry by-product meal (106 isolates). Thirty-seven isolates were resistant to at least three antimicrobial classes. Class 1 integrons were detected in 26 isolates, and the analysis of the variable region between the 5' conserved segment (CS) and 3' CS of each class 1 integron-positive isolate showed that 13 contained a typical 3' CS and 14 contained an atypical 3' CS. One Salmonella Senftenberg isolate harbored two class 1 integrons, showing both typical and atypical 3' CSs. The highest percentage of resistance was found to sulfonamides, and sul genes were detected in the majority of the resistant isolates. Aminoglycoside resistance was detected in 50 isolates, and aadA and aadB were present in 28 and 32 isolates, respectively. In addition, strA and strB were detected in 78.1 and 65.6% isolates resistant to streptomycin, respectively. Twenty-one isolates presented reduced susceptibility to β-lactams and harbored bla(TEM), bla(CMY), and/or bla(CTX-M). Forty isolates showed reduced susceptibility to tetracycline, and most presented tet genes. These results highlight the importance of the environment as a reservoir of resistant Salmonella, which may enable the persistence of resistance determinants in the poultry production chain, contributing, therefore, to the debate regarding the impacts that antimicrobial use in animal production may exert in human health.

  15. Identification and characterization of acyclovir-resistant clinical HSV-1 isolates from children.

    Science.gov (United States)

    Wang, Yi; Wang, Qi; Zhu, Qinchang; Zhou, Rong; Liu, Jinsong; Peng, Tao

    2011-10-01

    The occurrence of herpes simplex virus (HSV) with acyclovir (ACV) resistance is a cause for concern due to the frequent use of ACV for treatment, suppressive therapy, and prophylaxis of HSV infection. Although HSV infection is prevalent among children, very little is known about the drug susceptibility of HSV circulating in this patient population. To determine the status of ACV resistant HSV-1 among children. A reporter cell-based HSV infection assay (mVILA) was developed to conveniently evaluate the ACV susceptibility of HSV-1 clinical strains and used to analyze 68 HSV-1 primary isolates from oral lesions in children. Compared with PRA, mVILA is easier to perform. Using mVILA, HSV-1 isolates C106, C153, and C174 were found completely resistant to ACV, with a greater than 100-fold increase in IC50s. Sequence analysis of thymidine kinase (TK) and DNA polymerase (DNA POL) genes identified 11 new mutations. Structural modeling of the TK and DNA POL proteins suggested structural changes that might alter their interactions with ACV and ACV triphosphate, respectively. The insertion of a single G in a seven-guanine homopolymeric repeat sequence generated a truncated TK protein in C106. This study provides preliminary data on the ACV susceptibility status of HSV-1 in children. The prevalence rate of ACV-resistant HSV-1 in children was higher than predicted. Moreover, multiple mechanisms leading to the resistance were identified. These results suggest that new anti-herpetics with different working mechanisms should be valuable. Copyright © 2011 Elsevier B.V. All rights reserved.

  16. [Antimicrobial Susceptibility and Resistance Mutations in Campylobacter jejuni and C. coli Isolates from Human and Meat Sources].

    Science.gov (United States)

    Oishi, Akira; Murakami, Koichi; Etoh, Yoshiki; Sera, Nobuyuki; Horikawa, Kazumi

    2015-03-01

    Recently, there has been a marked increase in the number of reports of fluoroquinolone-resistant Campylobacter jejuni and Campylobacter coli. The aim of this study was to evaluate the prevalence of antimicrobial resistance and its genetic determinants in Campylobacter species isolated from meat and human subjects in Fukuoka Prefecture, Japan. Between 2011 and 2013, 55 and 64 isolates were collected from meat (chicken meat and beef liver) and humans, respectively, in this prefecture. Antimicrobial susceptibility tests were conducted using the agar dilution method in accordance with the Clinical and Laboratory Standards Institute guidelines, using the following 11 antimicrobial agents : cephalexin, cefoxitin, nalidixic acid, ciprofloxacin, levofloxacin, tetracycline, minocycline, ampicillin, streptomycin, kanamycin and erythromycin. The susceptibility rates of the isolates to three quinolones (nalidixic acid, ciprofloxacin, levofloxacin) were 43.7%, 41.2%, 40.3%, respectively. All the isolates were multidrug resistant. Whereas 46.9%-51.6% of the human isolates were resistant to one or more of the quinolones, only 32.7%-34.5% of the meat isolates were resistant to one or more of the drugs. DNA sequencing showed that of the 50 quinolone resistant isolates 44 had position 86 isoleucine (Ile) substituted for threonine (Thr) in the GyrA protein (Thr86Ile). This amino acid substitution resulted from ACA to ATA and ACT to ATT mutations of codon 86 in C. jejuni and C. coli, respectively. Furthermore, two of the four C. jejuni isolates lacking the Thr86Ile mutation had combined Ser22Gly-Asn203Ser substitutions, while the remaining two isolates had combined Ser22Gly-Asn203Ser-Ala 206Val substitutions. These four isolates also had cmeABC sequences that differed from the quinolone sensitive C. jejuni ATCC33560(T) strain. In conclusion, C. jejuni and C. coli have relatively high quinolone resistance, and are resistant to other antibiotics. The new combination of amino acid

  17. Label-free SRM-based relative quantification of antibiotic resistance mechanisms in Pseudomonas aeruginosa clinical isolates

    Directory of Open Access Journals (Sweden)

    Yannick eCharretier

    2015-02-01

    Full Text Available Both acquired and intrinsic mechanisms play a crucial role in Pseudomonas aeruginosa antibiotic resistance. Many clinically relevant resistance mechanisms result from changes in gene expression, namely multidrug efflux pump overproduction, AmpC beta-lactamase induction or derepression, and inactivation or repression of the carbapenem-specific porin OprD. Changes in gene expression are usually assessed using reverse-transcription quantitative real-time PCR (RT-qPCR assays. Here, we evaluated label-free Selected Reaction Monitoring (SRM-based mass spectrometry to directly quantify proteins involved in antibiotic resistance. We evaluated the label-free SRM using a defined set of P. aeruginosa isolates with known resistance mechanisms and compared it against RT-qPCR. Referring to efflux systems, we found a more robust relative quantification of antibiotic resistance mechanisms by SRM than RT-qPCR. The SRM-based approach was applied to a set of clinical P. aeruginosa isolates to detect antibiotic resistance proteins. This multiplexed SRM-based approach is a rapid and reliable method for the simultaneous detection and quantification of resistance mechanisms and we demonstrate its relevance for antibiotic resistance prediction.

  18. Antibiotic resistant coliform bacilli, isolated from freshly slaughtered poultry and from chilled poultry at retail outlets.

    Science.gov (United States)

    Bensink, J C; Botham, F P

    1983-03-01

    Antibiotic resistance in coliforms isolated from poultry was investigated. Poultry carcases were examined immediately after slaughter or at retail outlets; the carcases were from the same processing plant and 100 were examined from each source. Approximately 85% of the total of 13,858 isolates examined were found to be resistant to at least one antibiotic. Highly significant differences were found in the levels of antibiotic resistance from the 2 sources; ampicillin, chloramphenicol, and sulphonamide resistance was found more frequently in isolates from poultry at retail, while resistance against streptomycin and neomycin occurred more frequently in isolates from poultry examined at slaughter. The data were insufficient to explain these changes. Transfer of resistance occurred less frequently in isolates from poultry at retail; in particular the transfer of resistance from coliforms other than Escherichia coli was found to be greatly reduced.

  19. The tetracycline resistance determinant Tet 39 and the sulphonamide resistance gene sulII are common among resistant Acinetobacter spp. isolated from integrated fish farms in Thailand

    DEFF Research Database (Denmark)

    Agersø, Yvonne; Petersen, Andreas

    2007-01-01

    Objectives: To determine the genetic basis for tetracycline and sulphonamide resistance and the prevalence of class I and II integrons in oxytetracycline-resistant Acinetobacter spp. from integrated fish farms in Thailand. Methods: A total of 222 isolates were screened for tetracycline resistance...... genes [tet(A), tet(B), tet(H), tet(M) and tet(39)] and class II integrons by PCR. One hundred and thirty-four of these isolates were also sulphonamide resistant and these isolates were screened for sulphonamide resistance genes (sulII and sulIII) as well as class I integrons. Plasmid extraction...

  20. Antimicrobial susceptibility of Neisseria gonorrhoeae isolates from Hefei (2014?2015): genetic characteristics of antimicrobial resistance

    OpenAIRE

    Jiang, Fa-Xing; Lan, Qian; Le, Wen-Jing; Su, Xiao-Hong

    2017-01-01

    Background Antimicrobial resistance (AMR) and genetic determinants of resistance of N. gonorrhoeae isolates from Hefei, China, were characterized adding a breadth of information to the molecular epidemiology of gonococcal resistance in China. Methods 126?N. gonorrhoeae isolates from a hospital clinic in Hefei, were collected between January, 2014, and November, 2015. The minimum inhibitory concentration (MIC) of N. gonorrhoeae isolates for seven antimicrobials were determined by the agar dilu...

  1. EMERGENCE OF MULTI DRUG RESISTANT STRAINS OF E. COLI ISOLATED FROM URINARY TRACT INFECTION IN NAMAKKAL

    OpenAIRE

    2012-01-01

    In this study we evaluated the multi drug resistant strains of Escherichia coli isolated from urinary tract infection in different age groups in Namakkal. Totally 153 isolates of E.coli were obtained from 700 urine samples. Present study indicates that there is a high prevalence of multidrug resistant E. coli in female 24.34% followed by male 18.14%. After confirmation, isolates were performed to resistance patterns of different antibiotics were determined by standard disk diffusion method. T...

  2. Fluoroquinolone-Resistant Haemophilus parasuis Isolates Exhibit More Putative Virulence Factors than Their Susceptible Counterparts

    OpenAIRE

    Zhang, Qiang; Liu, Jiantao; Yan, Shuxian; Yang, Yujie; Zhang, Anding; Jin, Meilin

    2013-01-01

    The prevalence of 23 putative virulence factors among fluoroquinolone-susceptible and -resistant Haemophilus parasuis isolates was analyzed. Putative hemolysin precursor, fimbrial assembly chaperone, and type I site-specific restriction modification system R subunit genes were more prevalent among fluoroquinolone-resistant H. parasuis isolates than among fluoroquinolone-susceptible H. parasuis isolates. Fluoroquinolone resistance may be associated with an increase in the presence of some viru...

  3. Isolation of vancomycin resistant Enterococcus faecium from food

    DEFF Research Database (Denmark)

    Wegener, Henrik Caspar; Madsen, Mogens; Nielsen, Niels

    1997-01-01

    In a survey of vancomycin resistant Enterococcus faecium (VREF) in Danish meat products, VREF could be detected in 16% of 160 samples of broilers collected at slaughterhouses and in 15% of 26 samples of pork collected from the retail trade. VREF were isolated by enrichment for 24 h in nutrient...... was not detected in 124 samples of pork and 128 samples of beef from retail outlets by the direct plating method. An additional enrichment step in nutrient broth supplemented with vancomycin enhanced the detection rate of VREF by approximately three times compared to the direct plating method when investigating...... the same 160 samples of broilers by the two methods. The implications and public health aspects of VREF in food is discussed....

  4. Diversity of mercury resistance determinants among Bacillus strains isolated from sediment of Minamata Bay.

    Science.gov (United States)

    Narita, Masaru; Chiba, Kazuyuki; Nishizawa, Hiroshi; Ishii, Hidenori; Huang, Chieh-Chen; Kawabata, Zen'ichiro; Silver, Simon; Endo, Ginro

    2003-06-06

    Thirty mercury-resistant (Hg R) Bacillus strains were isolated from mercury-polluted sediment of Minamata Bay, Japan. Mercury resistance phenotypes were classified into broad-spectrum (resistant to inorganic Hg(2+) and organomercurials) and narrow-spectrum (resistant to inorganic Hg(2+) and sensitive to organomercurials) groups. Polymerase chain reaction (PCR) product sizes and the restriction nuclease site maps of mer operon regions from all broad-spectrum Hg R Bacillus were identical to that of Bacillus megaterium MB1. On the other hand, the PCR products of the targeted merP (extracellular mercury-binding protein gene) and merA (intracellular mercury reductase protein gene) regions from the narrow-spectrum Hg R Bacillus were generally smaller than those of the B. megaterium MB1 mer determinant. Diversity of gene structure configurations was also observed by restriction fragment length polymorphism (RFLP) profiles of the merA PCR products from the narrow-spectrum Hg R Bacillus. The genetic diversity of narrow-spectrum mer operons was greater than that of broad-spectrum ones.

  5. Identification of antimony resistance markers in Leishmania tropica field isolates through a cDNA-AFLP approach.

    Science.gov (United States)

    Kazemi-Rad, Elham; Mohebali, Mehdi; Khadem-Erfan, Mohammad Bagher; Saffari, Mojtaba; Raoofian, Reza; Hajjaran, Homa; Hadighi, Ramtin; Khamesipour, Ali; Rezaie, Sassan; Abedkhojasteh, Hoda; Heidari, Mansour

    2013-10-01

    Pentavalent antimonial compounds have been the first line therapy for leishmaniasis; unfortunately the rate of treatment failure of anthroponotic cutaneous leishmaniasis (ACL) is increasing due to emerging of drug resistance. Elucidation of the molecular mechanisms operating in antimony resistance is critical for development of new strategies for treatment. Here, we used a cDNA-AFLP approach to identify gene(s) which are differentially expressed in resistant and sensitive Leishmania tropica field isolates. We identified five genes, aquaglyceroporin (AQP1) acts in drug uptake, ATP-binding cassette (ABC) transporter (MRPA) involved in sequestration of drug, phosphoglycerate kinase (PGK) implicated in glycolysis metabolism, mitogen activated protein kinase (MAPK) and protein tyrosine phosphatase (PTP) responsible for phosphorylation pathway. The results were confirmed using real time RT-PCR which revealed an upregulation of MRPA, PTP and PGK genes and downregulation of AQP1 and MAPK genes in resistant isolate. To our knowledge, this is the first report of identification of PTP and PGK genes potentially implicated in resistance to antimonials. Our findings support the idea that distinct biomolecules might be involved in antimony resistance in L. tropica field isolates.

  6. Insertion sequence IS256 in canine pyoderma isolates of Staphylococcus pseudintermedius associated with antibiotic resistance.

    Science.gov (United States)

    Casagrande Proietti, P; Bietta, A; Coletti, M; Marenzoni, M L; Scorza, A V; Passamonti, F

    2012-06-15

    Staphylococcus pseudintermedius is the most frequent staphylococcal species isolated from canine pyoderma. The control of S. pseudintermedius infection is often difficult due to the expanded antimicrobial resistance phenotypes. Antibiotic resistance in staphylococcal pathogens is often associated to mobile genetic elements such as the insertion sequence IS256 that was first described as a part of the transposon Tn4001, which confers aminoglycoside resistance in Staphylococcus aureus and in Staphylococcus epidermidis. In this study a collection of 70 S. pseudintermedius isolates from canine pyoderma was used to investigate antimicrobial susceptibility to 15 antibiotics and the presence of IS256, not revealed in S. pseudintermedius yet. Antibiotic resistance profiling demonstrated that all S. pseudintermedius isolates had a multi-drug resistance phenotype, exhibiting simultaneous resistance to at least five antibiotics; indeed methicillin resistant S. pseudintermedius isolates were simultaneously resistant to at least nine antibiotics and all were also gentamicin resistant. PCR analyses revealed the presence of IS256 in 43/70 S. pseudintemedius isolates. The association between the presence of IS256 and the resistance was particularly significant for certain antibiotics: cefovecin, amikacin, gentamicin and oxacillin (χ(2)p-valuepseudintermedius isolates and its association with antibiotic resistance. Our findings suggest that S. pseudintermedius may acquire antibiotic resistance genes through mobile genetic elements which may play a predominant role in the dissemination of multi-drug resistance.

  7. Detection of antibiotic resistance and tetracycline resistance genes in Enterobacteriaceae isolated from the Pearl rivers in South China

    Energy Technology Data Exchange (ETDEWEB)

    Tao Ran [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, 511 Kehua Street, Tianhe District, Guangzhou 510640 (China); Ying Guangguo, E-mail: guangguo.ying@gmail.co [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, 511 Kehua Street, Tianhe District, Guangzhou 510640 (China); Su Haochang [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, 511 Kehua Street, Tianhe District, Guangzhou 510640 (China); Zhou Hongwei [Department of Environmental Health, School of Public Health and Tropical Medicine, Southern Medical University, 1838 North Guangzhou Street, Baiyun District, Guangzhou 510515 (China); Sidhu, Jatinder P.S. [CSIRO Land and Water, Queensland Bioscience Precinct, 306 Carmody Road, St Lucia QLD 4067 (Australia)

    2010-06-15

    This study investigated antibiotic resistance profiles and tetracycline resistance genes in Enterobacteriaceae family isolates from the Pearl rivers. The Enterobacteriaceae isolates were tested for susceptibility to seven antibiotics ampicillin, chloramphenicol, ciprofloxacin, levofloxacin, sulphamethoxazole/trimethoprim, tetracycline and trimethoprim. In Liuxi reservoir, with an exception to ampicillin resistant strains (11%) no other antibiotic resistance bacterial strains were detected. However, multiple drug resistance in bacterial isolates from the other sites of Pearl rivers was observed which is possibly due to sewage discharge and input from other anthropogenic sources along the rivers. Four tetracycline resistance genes tet A, tet B, tet C and tet D were detected in the isolates from the rivers. The genes tet A and tet B were widely detected with the detection frequencies of 43% and 40% respectively. Ciprofloxacin and levofloxacin resistant enteric bacteria were also isolated from the pig and duck manures which suggest a wider distribution of human specific drugs in the environment. This investigation provided a baseline data on antibiotic resistance profiles and tetracycline resistance genes in the Pearl rivers delta. - High rates of antibiotic resistance in Enterobacteriaceae from river water are attributed to wastewater contamination.

  8. Comparison of protein profiles of beech bark disease-resistant or beech bark disease-susceptible American beech

    Science.gov (United States)

    Mary E. Mason; Marek Krasowski; Judy Loo; Jennifer. Koch

    2011-01-01

    Proteomic analysis of beech bark proteins from trees resistant and susceptible to beech bark disease (BBD) was conducted. Sixteen trees from eight geographically isolated stands, 10 resistant (healthy) and 6 susceptible (diseased/infested) trees, were studied. The genetic complexity of the sample unit, the sampling across a wide geographic area, and the complexity of...

  9. Plasmid-mediated quinolone resistance among non-typhi Salmonella enterica isolates, USA

    Science.gov (United States)

    We determined the prevalence of plasmid-mediated quinolone resistance mechanisms among non-Typhi Salmonella (NTS) spp. isolates from humans, food animals, and retail meat in the United States in 2007. Fifty-one (2.4%) of human isolates (n=2165), 5 (1.6%) of isolates from animal isolates (n=1915) an...

  10. Antibiotic Resistance in Staphylococcus aureus and Coagulase Negative Staphylococci Isolated from Goats with Subclinical Mastitis

    Directory of Open Access Journals (Sweden)

    Salvatore Virdis

    2010-01-01

    Full Text Available Antimicrobial resistance patterns and gene coding for methicillin resistance (mecA were determined in 25 S. aureus and 75 Coagulase Negative Staphylococci (CNS strains isolates from half-udder milk samples collected from goats with subclinical mastitis. Fourteen (56.0% S. aureus and thirty-one (41.3% CNS isolates were resistant to one or more antimicrobial agents. S. aureus showed the highest resistance rate against kanamycin (28.0%, oxytetracycline (16.0%, and ampicillin (12.0%. The CNS tested were more frequently resistant to ampicillin (36.0% and kanamycin (6.7%. Multiple antimicrobial resistance was observed in eight isolates, and one Staphylococcus epidermidis was found to be resistant to six antibiotics. The mecA gene was not found in any of the tested isolates. Single resistance against β-lactamics or aminoglicosides is the most common trait observed while multiresistance is less frequent.

  11. Isolation and identification of antibiotic resistance genes in Staphylococcus aureus isolates from respiratory system infections in shahrekord, Iran

    Directory of Open Access Journals (Sweden)

    Maryam Reisi

    2014-07-01

    Full Text Available   Introduction : Staphylococcus aureus is considered as one of pathogenic agents in humans, that engages different body parts including respiratory system and causes to spend lots of costs and extending patient’s treatment period. This study which is performed to separate and investigate the pattern of antibiotic resistance in Staphylococcus aureus isolates from upper respiratory system infections in Shahrekord.   Materials and methods: This study was done by sectional-descriptive method On 200 suspicious persons to the upper respiratory system infections who were referred to the Imam Ali clinic in Shahrekord in 2012. After isolation of Staphylococcus aureus from cultured nose discharges, antibiotic resistance genes were identified by polymerase chain reaction (PCR by using defined primer pairs .   Results : Among 200 investigated samples in 60 cases (30% Staphylococcus aureus infection (by culturing and PCR method was determined. Isolates showed the lowest amount of antibiotic resistance to vancomycin (0.5% and the highest amount of resistance to the penicillin G and cefotaxime (100%. mecA gene (encoding methicillin resistance with frequency of 85.18% and aacA-D gene (encoding resistance to aminoglycosides with frequency of 28.33% showed the highest and lowest frequency of antibiotic resistance genes coding in Staphylococcus aureus isolates respectively .   Discussion and conclusion : Notable prevalence of resistant Staphylococcus aureus isolates in community acquired respiratory infections, recommend continuous control necessity to impede the spreading of these bacteria and their infections.  

  12. Occurrence Of Virulence Factors And Antimicrobial Resistance In Pasteurella Multocida Strains Isolated From Slaughter Cattle In Iran

    Directory of Open Access Journals (Sweden)

    Faham eKhamesipour

    2014-10-01

    Full Text Available A total of 30 Pasteurella multocida strains isolated from 333 pneumonic and apparently health slaughter cattle were examined for capsule biosynthesis genes and 23 virulence associated genes by polymerase chain reaction. The disc diffusion technique was used to determine antimicrobial resistance profiles among the isolates. Of the isolates, 23 belonged to capsular type A, 5 to capsular type D and two isolates were untypeable. The distribution of the capsular types in pneumonic lungs and in apparently health lungs was statistically similar. All virulence genes tested were detected among the isolates derived from pneumonic lungs; whereas isolates derived from apparently health lungs carried 16 of the 23 genes. The frequently detected genes among isolates from pneumonic lungs were exbD, hgbA, hgbB, ompA, ompH, oma87 and sodC; whereas tadD, toxA and pmHAS genes occurred less frequently. Most of the adhesins and superoxide dismutases; and all of the iron acquisition and protectin proteins occurred at significantly (p≤0.05 higher frequencies in isolates from pneumonic lungs. Isolates from apparently healthy lungs didn’t carry the following genes; hsf-1, hsf-2, tadD, toxA, nanB, nanH and pmHAS. One adhesion (hsf-1 and two iron acquisition (exbD and tonB genes occurred at significantly (p≤0.05 higher frequencies among capA isolates. All the P. multocida isolates were susceptible to ciprofloxacin, co-trimoxazole, doxycycline, enrofloxacin, nitrofurantoin and tetracyclines. Different proportions of the isolates were however resistant to ampicillin, amoxicillin, erythromycin, lincomycin, penicillin, rifampin, streptomycin and florfenicol. Our results reveal presence of virulence factors in P. multocida strains isolated from symptomatic and asymptomatic bovids. A higher frequency of the factors among isolates from symptomatic study animals may suggest their role in pathogenesis of P. multocida-associated bovine respiratory disease. The results further

  13. Antimicrobial Resistance among Salmonella and Shigella Isolates in Five Canadian Provinces (1997 to 2000

    Directory of Open Access Journals (Sweden)

    Leah J Martin

    2006-01-01

    Full Text Available OBJECTIVE: To describe rates of antimicrobial resistance (AMR among Salmonella and Shigella isolates reported in five Canadian provinces, focusing on clinically important antimicrobials.

  14. Determination of Ampicillin Resistant Enterococci (ARE Isolated From Canine and Feline Rectal Swabs

    Directory of Open Access Journals (Sweden)

    Baran CELIK

    2017-01-01

    Full Text Available Enterococci species, which are normal inhabitants of the gut flora of healthy animals and human, began to be recognized as an important pathogen in both human and veterinary medicine due to the acquired resistance profiles. The aim of the study is to examine the diversity of ampicillin resistance enterococci (ARE species in cats and dogs, their antimicrobial susceptibility profiles and to determine some of the virulence related genes; ace, gelE, efaA, agg and esp. For this purpose, rectal swabs from companion animals were collected and processed for ampicillin resistant enterococci isolation. One hundred fifty seven swab samples (86 canine and 71 feline were examined. ARE were isolated from 18 canine and 18 feline samples. All isolates identified as E. faecium by PCR. Antimicrobial susceptibilities of the isolates were determined by disk diffusion method. The isolates were resistant to ampicillin, penicillin, tetracycline (100%, followed by rifampicin and erythromycin (97%, streptomycin (92%, gentamicin (81%, ciprofloxacin (61%, nitrofurantoin (19%. Only two of E. faecium isolates were resistant to vancomycin and one to chloramphenicol. Multidrug resistance (resistance ≥4 antimicrobials observed in all isolates. Virulence genes ace, agg and esp were not detected in any of the tested isolates. The efaA and gelE genes detection rates were, 13.8% and 11.1% respectively. The ARE isolation rate among pet animals was 22.9%. Screening of antimicrobial resistant enterococci among companion animals would be useful to detect any emerging antimicrobial resistance problem related with public health.

  15. Functional properties of proteins isolated from industrially produced sunflower meal

    Directory of Open Access Journals (Sweden)

    Petia Ivanova

    2014-10-01

    Full Text Available Protein isolate 1 (PI1 and protein isolate 2 (PI2 were prepared from industrially produced sunflower meal by using isoelectric and ethanol precipitation respectively. The water absorption capacity of PI1 was 6 times higher than that of PI2 and was significantly reduced by the presence of 0.03 M and 0.25 M NaCl. Oil absorption capacity of both protein isolates was not influenced by NaCl supplementation. Foam capacity of PI1 and PI2 was pH-dependent. While the foam capacity of both isolates was improved by either 0.03 M or 0.25 M NaCl, the foam stability was negatively influenced by the addition of NaCl at all pH values with except for pH 4. Emulsifying activity of PI1 and PI2 was lowest at pH 4. The emulsions exhibited relatively high stability (> 90% under all studied conditions. Knowledge of the influence of pH and boundary concentrations of NaCl on the functionality of sunflower meal protein isolates could be beneficial for their future potential application in food industry.

  16. Draft Genome Sequences of Pandrug-Resistant Serratia marcescens Clinical Isolates Harboring blaNDM-1

    Science.gov (United States)

    Yao, Yancheng; Falgenhauer, Linda; Kempf, Volkhard A. J.; Hogardt, Michael; Göttig, Stephan; Imirzalioglu, Can

    2017-01-01

    ABSTRACT The draft genome sequences of two clonal, pandrug-resistant Serratia marcescens clinical isolates were determined. The resistance phenotype was plasmid driven, as 14 of 17 resistance genes were present on large IncFIB(K), IncHI2, and IncA/C2 plasmids indicating a large pool of transmissible antibiotic resistance genes. PMID:28104656

  17. Genome Sequences of Two Copper-Resistant Escherichia coli Strains Isolated from Copper-Fed Pigs

    DEFF Research Database (Denmark)

    Lüthje, Freja L.; Hasman, Henrik; Aarestrup, Frank Møller;

    2014-01-01

    The draft genome sequences of two copper-resistant Escherichia coli strains were determined. These had been isolated from copper-fed pigs and contained additional putative operons conferring copper and other metal and metalloid resistances.......The draft genome sequences of two copper-resistant Escherichia coli strains were determined. These had been isolated from copper-fed pigs and contained additional putative operons conferring copper and other metal and metalloid resistances....

  18. Antimicrobial resistance of Escherichia coli isolated from chickens with colibacillosis in and around Harare, Zimbabwe.

    Science.gov (United States)

    Saidi, Bamusi; Mafirakureva, Prettimore; Mbanga, Joshua

    2013-03-01

    Colibacillosis, a disease caused by avian pathogenic Escherichia coli (APEC), can lead to great economic losses in the poultry industry. The aim of this study was to determine the prevalence of antibiotic resistance and antibiotic resistance patterns in APEC in Zimbabwe. From 503 chickens diagnosed with colibacillosis, 103 E. coli isolates were obtained. Isolation and identification of E. coli were carried out using microscopy and biochemical tests. The disc diffusion method was used to determine antibiotic susceptibility of the isolates to 8 commercial antibiotics. Many isolates exhibited resistance to more than one antibiotic. Antibiogram profiles indicated maximum resistance to tetracycline (100%), bacitracin (100%), and cloxacillin (100%) and a high prevalence of resistance to ampicillin (94.1%). However; there were high prevalences of sensitivity to ciprofloxacin (100%) and gentamycin (97.1%). The isolates showed moderate rates of sensitivity to chloramphenicol and neomycin. All isolates in this study showed multidrug resistance because they were all resistant to 3 or more antibiotics. Seven multidrug resistance patterns were observed. The most common pattern (resistance to ampicillin, bacitracin, cloxacillin, and tetracycline) was exhibited by 30 isolates. Our findings show that there is emerging drug resistance in APEC associated with colibacillosis in Zimbabwe. The observed high level of multidrug resistance could hamper the treatment of colibacillosis in Zimbabwe.

  19. Towards the Understanding of Resistance Mechanisms in Clinically Isolated Trimethoprim-resistant, Methicillin-resistant Staphylococcus aureus Dihydrofolate Reductase

    Energy Technology Data Exchange (ETDEWEB)

    Frey, K.; Lombardo, M; Wright, D; Anderson, A

    2010-01-01

    Resistance to therapeutics such as trimethoprim-sulfamethoxazole has become an increasing problem in strains of methicillin-resistant Staphylococcus aureus (MRSA). Clinically isolated trimethoprim-resistant strains reveal a double mutation, H30N/F98Y, in dihydrofolate reductase (DHFR). In order to develop novel and effective therapeutics against these resistant strains, we evaluated a series of propargyl-linked antifolate lead compounds for inhibition of the mutant enzyme. For the propargyl-linked antifolates, the F98Y mutation generates minimal (between 1.2- and 6-fold) losses of affinity and the H30N mutation generates greater losses (between 2.4- and 48-fold). Conversely, trimethoprim affinity is largely diminished by the F98Y mutation (36-fold) and is not affected by the H30N mutation. In order to elucidate a mechanism of resistance, we determined a crystal structure of a complex of this double mutant with a lead propargyl-linked antifolate. This structure suggests a resistance mechanism consistent both for the propargyl-linked class of antifolates and for trimethoprim that is based on the loss of a conserved water-mediated hydrogen bond.

  20. Increasing antibiotic resistance in preservative-tolerant bacterial strains isolated from cosmetic products.

    Science.gov (United States)

    Orús, Pilar; Gomez-Perez, Laura; Leranoz, Sonia; Berlanga, Mercedes

    2015-03-01

    To ensure the microbiological quality, consumer safety and organoleptic properties of cosmetic products, manufacturers need to comply with defined standards using several preservatives and disinfectants. A drawback regarding the use of these preservatives is the possibility of generating cross-insusceptibility to other disinfectants or preservatives, as well as cross resistance to antibiotics. Therefore, the objective of this study was to understand the adaptive mechanisms of Enterobacter gergoviae, Pseudomonas putida and Burkholderia cepacia that are involved in recurrent contamination in cosmetic products containing preservatives. Diminished susceptibility to formaldehyde-donors was detected in isolates but not to other preservatives commonly used in the cosmetics industry, although increasing resistance to different antibiotics (β-lactams, quinolones, rifampicin, and tetracycline) was demonstrated in these strains when compared with the wild-type strain. The outer membrane protein modifications and efflux mechanism activities responsible for the resistance trait were evaluated. The development of antibiotic-resistant microorganisms due to the selective pressure from preservatives included in cosmetic products could be a risk for the emergence and spread of bacterial resistance in the environment. Nevertheless, the large contribution of disinfection and preservation cannot be denied in cosmetic products. Copyright© by the Spanish Society for Microbiology and Institute for Catalan Studies.

  1. Vancomycin modifies the expression of the agr system in multidrug-resistant Staphylococcus aureus clinical isolates

    Directory of Open Access Journals (Sweden)

    Vicenta eCázares-Domínguez

    2015-05-01

    Full Text Available Staphylococcus aureus is an opportunistic pathogen that colonizes human hosts and causes a wide variety of diseases. Two interacting regulatory systems called agr (accessory gene regulator and sar (staphylococcal accessory regulator are involved in the regulation of virulence factors. The aim of this study was to evaluate the effect of vancomycin on hld and spa gene expression during the exponential and post-exponential growth phases in multidrug resistant (MDR S. aureus. Methods. Antibiotic susceptibility was evaluated by the standard microdilution method. The phylogenetic profile was obtained by pulsed-field gel electrophoresis. Polymorphisms of agr and SCCmec were analyzed by multiplex polymerase chain reaction. The expression levels of hld and spa were analyzed by reverse transcription-PCR. An enzyme-linked immunosorbent assay assay was performed to detect protein A, and biofilm formation was analyzed via crystal violet staining. Results. In total, 60.60% (20/33 of S. aureus clinical isolates were MDR. Half (10/20 of the MDR S. aureus isolates were distributed in subcluster 10, with > 90% similarity among them. In the isolates of this subcluster, a high prevalence (100% for the agrII and the cassette SCCmec II polymorphisms was found. Our data showed significant increases in hld expression during the post-exponential phase in the presence and absence of vancomycin. Significant increases in spa expression, protein A production and biofilm formation were observed during the post-exponential phase when the MDR S. aureus isolates were challenged with vancomycin. Conclusion. The polymorphism agrII, which is associated with nosocomial isolates, was the most prevalent polymorphism in MDR S. aureus. Additionally, under our study conditions, vancomycin modified hld and spa expression in these clinical isolates. Therefore, vancomycin may regulate alternative systems that jointly participate in the regulation of these virulence factors.

  2. The spa typing of methicillin-resistant Staphylococcus aureus isolates by High Resolution Melting (HRM) analysis.

    Science.gov (United States)

    Fasihi, Yasser; Fooladi, Saba; Mohammadi, Mohammad Ali; Emaneini, Mohammad; Kalantar-Neyestanaki, Davood

    2017-09-06

    Molecular typing is an important tool for control and prevention of infection. A suitable molecular typing method for epidemiological investigation must be easy to perform, highly reproducible, inexpensive, rapid and easy to interpret. In this study, two molecular typing methods including the conventional PCR-sequencing method and high resolution melting (HRM) analysis were used for staphylococcal protein A (spa) typing of 30 Methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered from clinical samples. Based on PCR-sequencing method results, 16 different spa types were identified among the 30 MRSA isolates. Among the 16 different spa types, 14 spa types separated by HRM method. Two spa types including t4718 and t2894 were not separated from each other. According to our results, spa typing based on HRM analysis method is very rapid, easy to perform and cost-effective, but this method must be standardized for different regions, spa types, and real-time machinery.

  3. Antibiotic resistance patterns of Pseudomonas spp. isolated from the River Danube

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    Clemens eKittinger

    2016-05-01

    Full Text Available Spread and persistence of antibiotic resistance pose a severe threat to human health, yet there is still lack of knowledge about reservoirs of antibiotic resistant bacteria in the environment. We took the opportunity of the Joint Danube Survey 3 (JDS3, the world's biggest river research expedition of its kind in 2013, to analyse samples originating from different sampling points along the whole length of the river. Due to its high clinical relevance, we concentrated on the characterization of Pseudomonas spp. and evaluated the resistance profiles of Pseudomonas spp. which were isolated from eight sampling points. In total, 520 Pseudomonas isolates were found, 344 (66.0% isolates were identified as Pseudomonas putida, and 141 (27.1% as Pseudomonas fluorescens, all other Pseudomonas species were represented by less than five isolates, among those two P. aeruginosa isolates. Thirty seven percent (37% of all isolated Pseudomonas species showed resistance to at least one out of eleven tested antibiotics. The most common resistance was against meropenem (30.4% / 158 isolates piperacillin/tazobactam (10.6% / 55 isolates and ceftazidime (4.2% / 22 isolates. 16 isolates (3.1% / 16 isolates were multi-resistant. For each tested antibiotic at least one resistant isolate could be detected. Sampling points from the upper stretch of the River Danube showed more resistant isolates than downriver. Our results suggest that antibiotic resistance can be acquired by and persists even in Pseudomonas species that are normally not in direct contact with humans. A possible scenario is that these bacteria provide a reservoir of antibiotic resistance genes that can spread to related human pathogens by horizontal gene transfer.

  4. Isolation and characterization of selenate resistant mutants of Acremonium chrysogenum

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    Airton Vialta

    1999-01-01

    Full Text Available Mutants unable to convert exogenous sulfate to sulfite were isolated using the toxic analogue selenate. Three of twenty-eight isolated mutants were chromate sensitive. They showed a possible lesion in the gene that codes the ATP sulfurylase. The others were chromate resistant, and probably had a lesion in one or both of the genes that code the sulfate permease. Methionine increased the resistance levels to selenate. In addition, the frequency of spontaneous mutants obtained in a medium containing methionine was higher (between 2.4 x 10-6 and 18.0 x 10-6 than that obtained using a medium without any intentional source of sulfur (between 0.7 x 10-6 and 5.0 x 10-6. The original strain, as well as the mutants, were able to grow in a sulfur-free liquid medium even after 4 consecutive inoculation procedures. These results indicated the existence of sulfur traces in the medium and/or an efficient intracellular storage system. There was no significant difference between cephalosporin C production in mutants and the original strain.Mutantes incapazes de converter o sulfato extracelular em sulfito foram isolados utilizando o análogo tóxico selenato. De 28 mutantes isolados, apenas 3 foram sensíveis ao cromato, provavelmente apresentando lesão no gene que codifica a ATP sulfurilase. Os demais foram resistentes ao cromato e devem conter lesão no gene sB ou também no gene sC. A metionina elevou os níveis de resistência ao selenato e a freqüência de mutantes espontâneos obtida em meio contendo este aminoácido foi maior (entre 2,42 x 10-6 e 18,04 x 10-6 do que a obtida no meio sem a adição de qualquer fonte intencional de enxofre (entre 0,71 x 10-6 e 5,0 x 10-6. A linhagem original e os mutantes foram capazes de crescer, mesmo depois de quatro etapas de inóculo, fato que pode ser explicado pela existência de traços do referido elemento no meio e/ou a presença de um sistema eficiente de estocagem intracelular. A produção de cefalosporina C

  5. Isolation and characterization of a bacteriophage phiEap-2 infecting multidrug resistant Enterobacter aerogenes.

    Science.gov (United States)

    Li, Erna; Wei, Xiao; Ma, Yanyan; Yin, Zhe; Li, Huan; Lin, Weishi; Wang, Xuesong; Li, Chao; Shen, Zhiqiang; Zhao, Ruixiang; Yang, Huiying; Jiang, Aimin; Yang, Wenhui; Yuan, Jing; Zhao, Xiangna

    2016-06-20

    Enterobacter aerogenes (Enterobacteriaceae) is an important opportunistic pathogen that causes hospital-acquired pneumonia, bacteremia, and urinary tract infections. Recently, multidrug-resistant E. aerogenes have been a public health problem. To develop an effective antimicrobial agent, bacteriophage phiEap-2 was isolated from sewage and its genome was sequenced because of its ability to lyse the multidrug-resistant clinical E. aerogenes strain 3-SP. Morphological observations suggested that the phage belongs to the Siphoviridae family. Comparative genome analysis revealed that phage phiEap-2 is related to the Salmonella phage FSL SP-031 (KC139518). All of the structural gene products (except capsid protein) encoded by phiEap-2 had orthologous gene products in FSL SP-031 and Serratia phage Eta (KC460990). Here, we report the complete genome sequence of phiEap-2 and major findings from the genomic analysis. Knowledge of this phage might be helpful for developing therapeutic strategies against E. aerogenes.

  6. Evolutionary analysis of tomato Sw-5 resistance-breaking isolates of Tomato spotted wilt virus.

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    López, Carmelo; Aramburu, José; Galipienso, Luis; Soler, Salvador; Nuez, Fernando; Rubio, Luis

    2011-01-01

    Tomato spotted wilt virus (TSWV) causes severe economic losses in many crops worldwide and often overcomes resistant cultivars used for disease control. Comparison of nucleotide and amino acid sequences suggested that tomato resistance conferred by the gene Sw-5 can be overcome by the amino acid substitution C to Y at position 118 (C118Y) or T120N in the TSWV movement protein, NSm. Phylogenetic analysis revealed that substitution C118Y has occurred independently three times in the studied isolates by convergent evolution, whereas the substitution T120N was a unique event. Analysis of rates of non-synonymous and synonymous changes at individual codons showed that substitution C118Y was positively selected.

  7. Antibiotic Resistance Pattern and Prophage Typing of Methicillin Resistant Staphylococcus aureus Strains Isolated from Chicken Husbandries in Tehran

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    Fateh Rahimi

    2013-10-01

    Full Text Available Abstract Background and objective: Staphylococcus aureus is a common cause of infection among human and animals and known as community-acquired and nosocomial pathogen. Most of the isolates contain lysogenic phages which are responsible for production of various virulence factors. All staphylococcus isolates are classified in 6 groups according to their sensitivity to 27 known lysogenic phages. The aim of this study was to analyze different prophage types, antibiotic resistance pattern and detection of mecA gene in meticilin resistant S. aureus (MRSA strains isolated from a poultry farm in Karaj. Materials and methods: Totally 110 isolates of MRSA from poultry samples were collected from 1 laying flocks in Karaj. All isolates were identified at the species level using specific primers. Susceptibility to 19 antibiotics was determined using disc diffusion method according to guidelines of Clinical Laboratory and Standard institute (CLSI. Minimum inhibitory concentration (MIC of oxacillin and vancomycin in MRSA isolates were also detected using Etest according to CLSI recommendation. Primers for identification of 6 classes of prophages were used in a Multiplex-PCR assay. Plaque and spot assays were used to show the presence of lysogenic prophages. mecA gene was detected using specific primers. Results: Using PCR all isolates were confirmed as MRSA. The highest antibiotic resistance was observed to penicillin (100% and followed by ciprofloxacin (100%, erythromycin (100%, amikacin (100% and kanamycin (100%. None of the isolates were resistant to fusidic acid, nitrofurantoin, vancomycin, linezolid and synercid. Very high (MIC≥128 µg/ml levels resistance to oxacillin was observed in 83.5 % of the isolates. Four different prophage types were found in MRSA isolates. Out of the total MRSA isolates, 81 and 19 percent of the isolates contained 4 and 3 different prophage types, respectively. Conclusion: High diversity of prophage phages was found in MRSA

  8. Prevalence of vancomycin resistant Staphylococcus aureus (VRSA) in methicillin resistant S. aureus (MRSA) strains isolated from burn wound infections.

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    Hasan, Rashedul; Acharjee, Mrityunjoy; Noor, Rashed

    2016-01-01

    The increase in resistance of methicillin resistant Staphylococcus aureus (MRSA) strains to vancomycin has been perceived as a formidable threat in the therapeutic fields. The present study investigated the vancomycin resistance traits of MRSA isolates [vancomycin resistant S. aureus (VRSA)] collected from burn patients. Twenty-nine of 40 isolates of Staphylococcus spp. were identified as S. aureus which were further tested against 20 commercially available antibiotics to determine antibiotic susceptibility patterns. Imipenem was the most potential antibiotic resulting in 90% sensitivity, followed by netilmicin, clindamycin, and nitrofurantoin (80% sensitivity). All isolates were found to be resistant to penicillin. Approximately 75% of them were found to be resistant to methicillin, oxacillin, azithromycin, cipro-floxacin, and tetracycline. Approximately 45% isolates exhibited resistance to amikacin, chloramphenicol, gentamycin, and tobramycin. Twenty-one of the 29 strains of S. aureus were MRSA, of which 11 were resistant to vancomycin when employing the disc diffusion method. However, when the broth micro-dilution procedure was used to measure the minimum inhibitory concentration (MIC) of vancomycin, eight isolates were resistant to vancomycin, six with an MIC of 32 μg/mL and two with an MIC of 64 μg/mL. A significant fraction of VRSA was found among MRSA strains in this study, revealing the necessity for new and effective drugs against MRSA.

  9. Isolation of the intermediate filament protein vimentin by chromatofocusing.

    Science.gov (United States)

    Bloemendal, H; Willemsen, M; Groenewoud, G; Oomen, P

    1985-01-28

    A novel, simple and relatively rapid method is described for the isolation of the intermediate-sized filament protein vimentin from eye lens tissue. Chromatofocusing is applied as the sole purification step. The apparent isoelectric point of the protein in 6 M urea and at 22 degrees C is 4.9. Electrophoretic mobility on one- and two-dimensional polyacrylamide gels, solubility in 6 M urea and amino acid composition were used for identification.

  10. EMERGENCE OF MULTI DRUG RESISTANT STRAINS OF E. COLI ISOLATED FROM URINARY TRACT INFECTION IN NAMAKKAL

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    P. Poongothai

    2012-08-01

    Full Text Available In this study we evaluated the multi drug resistant strains of Escherichia coli isolated from urinary tract infection in different age groups in Namakkal. Totally 153 isolates of E.coli were obtained from 700 urine samples. Present study indicates that there is a high prevalence of multidrug resistant E. coli in female 24.34% followed by male 18.14%. After confirmation, isolates were performed to resistance patterns of different antibiotics were determined by standard disk diffusion method. The antibiogram patterns of the isolates showed a high percentage of multidrug resistance to cephodoxime, Novobiocin and vancomycin which was repeated to most of the isolates. Fifty resistance patterns were recognized among the MDR strains. This study revealed that E. coli was the predominant bacterial pathogen of community acquired UTIs in Aligarh, India. It also demonstrated an increasing resistance to number of antibiotics. This study is useful for clinician in order to improve the empirical treatment.

  11. Isolation and identification of cobalt- and caesium-resistant bacteria from a nuclear fuel storage pond.

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    Dekker, Linda; Osborne, Thomas H; Santini, Joanne M

    2014-10-01

    One of the issues facing the nuclear power industry is how to store spent nuclear fuel which is contaminated with radionuclides produced during nuclear fission, including caesium ((134)Cs(+), (135)Cs(+) and (137)Cs(+)) and cobalt ((60)Co(2+)). In this study, we have isolated Co(2+)- and Cs(+)-resistant bacteria from water collected from a nuclear fuel storage pond. The most resistant Cs(+) and Co(2+) isolates grew in the presence of 500 mM CsCl and 3 mM CoCl2. Strain Cs67-2 is resistant to fourfold more Cs(+) than Cupriavidus metallidurans str. CH34 making it the most Cs(+)-resistant strain identified to date. The Cs(+)-resistant isolates were closely related to bacteria in the Serratia and Yersinia genera, while the Co(2+)-resistant isolates were closely related to the Curvibacter and Tardiphaga genera. These new isolates could be used for bioremediation.

  12. Characterization of integrons and resistance genes in multidrug-resistant Salmonella enterica isolated from meat and dairy products in Egypt.

    Science.gov (United States)

    Ahmed, Ashraf M; Shimamoto, Toshi; Shimamoto, Tadashi

    2014-10-17

    Foodborne pathogens are a leading cause of illness and death, especially in developing countries. The problem is exacerbated if bacteria attain multidrug resistance. Little is currently known about the extent of antibiotic resistance in foodborne pathogens and the molecular mechanisms underlying this resistance in Africa. Therefore, the current study was carried out to characterize, at the molecular level, the mechanism of multidrug resistance in Salmonella enterica isolated from 1600 food samples (800 meat products and 800 dairy products) collected from different street venders, butchers, retail markets and slaughterhouses in Egypt. Forty-seven out of 69 isolates (68.1%) showed multidrug resistance phenotypes to at least three classes of antimicrobials. The incidence of multidrug-resistant isolates was higher in meat products (37, 69.8%) than in dairy products (10, 62.5%). The multidrug-resistant serovars included, S. enterica serovar Typhimurium (24 isolates, 34.8%), S. enterica serovar Enteritidis, (15 isolates, 21.8%), S. enterica serovar Infantis (7 isolates, 10.1%) and S. enterica non-typable serovar (1 isolate, 1.4%). The highest resistance was to ampicillin (95.7%), then to kanamycin (93.6%), spectinomycin (93.6%), streptomycin (91.5%) and sulfamethoxazole/trimethoprim (91.5%). PCR and DNA sequencing were used to screen and characterize integrons and antibiotic resistance genes and 39.1% and 8.7% of isolates were positive for class 1 and class 2 integrons, respectively. β-lactamase-encoding genes were identified in 75.4% of isolates and plasmid-mediated quinolone resistance genes were identified in 27.5% of isolates. Finally, the florphenicol resistance gene, floR, was identified in 18.8% of isolates. PCR screening identified S. enterica serovar Typhimurium DT104 in both meat and dairy products. This is the first study to report many of these resistance genes in dairy products. This study highlights the high incidence of multidrug-resistant S. enterica in

  13. Structure based in silico analysis of quinolone resistance in clinical isolates of Salmonella Typhi from India.

    Science.gov (United States)

    Kumar, Manoj; Dahiya, Sushila; Sharma, Priyanka; Sharma, Sujata; Singh, Tej P; Kapil, Arti; Kaur, Punit

    2015-01-01

    Enteric fever is a major cause of morbidity in several parts of the Indian subcontinent. The treatment for typhoid fever majorly includes the fluoroquinolone group of antibiotics. Excessive and indiscriminate use of these antibiotics has led to development of acquired resistance in the causative organism Salmonella Typhi. The resistance towards fluoroquinolones is associated with mutations in the target gene of DNA Gyrase. We have estimated the Minimum Inhibitory Concentration (MIC) of commonly used fluoroquinolone representatives from three generations, ciprofloxacin, ofloxacin, levofloxacin and moxifloxacin, for 100 clinical isolates of Salmonella Typhi from patients in the Indian subcontinent. The MICs have been found to be in the range of 0.032 to 8 μg/ml. The gene encoding DNA Gyrase was subsequently sequenced and point mutations were observed in DNA Gyrase in the quinolone resistance determining region comprising Ser83Phe/Tyr and Asp87Tyr/Gly. The binding ability of these four fluoroquinolones in the quinolone binding pocket of wild type as well as mutant DNA Gyrase was computationally analyzed by molecular docking to assess their differential binding behaviour. This study has revealed that mutations in DNA Gyrase alter the characteristics of the binding pocket resulting in the loss of crucial molecular interactions and consequently decrease the binding affinity of fluoroquinolones with the target protein. The present study assists in understanding the underlying molecular and structural mechanism for decreased fluoroquinolone susceptibility in clinical isolates as a consequence of mutations in DNA Gyrase.

  14. Structure Based In Silico Analysis of Quinolone Resistance in Clinical Isolates of Salmonella Typhi from India

    Science.gov (United States)

    Sharma, Priyanka; Sharma, Sujata; Singh, Tej P.; Kapil, Arti; Kaur, Punit

    2015-01-01

    Enteric fever is a major cause of morbidity in several parts of the Indian subcontinent. The treatment for typhoid fever majorly includes the fluoroquinolone group of antibiotics. Excessive and indiscriminate use of these antibiotics has led to development of acquired resistance in the causative organism Salmonella Typhi. The resistance towards fluoroquinolones is associated with mutations in the target gene of DNA Gyrase. We have estimated the Minimum Inhibitory Concentration (MIC) of commonly used fluoroquinolone representatives from three generations, ciprofloxacin, ofloxacin, levofloxacin and moxifloxacin, for 100 clinical isolates of Salmonella Typhi from patients in the Indian subcontinent. The MICs have been found to be in the range of 0.032 to 8 μg/ml. The gene encoding DNA Gyrase was subsequently sequenced and point mutations were observed in DNA Gyrase in the quinolone resistance determining region comprising Ser83Phe/Tyr and Asp87Tyr/Gly. The binding ability of these four fluoroquinolones in the quinolone binding pocket of wild type as well as mutant DNA Gyrase was computationally analyzed by molecular docking to assess their differential binding behaviour. This study has revealed that mutations in DNA Gyrase alter the characteristics of the binding pocket resulting in the loss of crucial molecular interactions and consequently decrease the binding affinity of fluoroquinolones with the target protein. The present study assists in understanding the underlying molecular and structural mechanism for decreased fluoroquinolone susceptibility in clinical isolates as a consequence of mutations in DNA Gyrase. PMID:25962113

  15. Structure based in silico analysis of quinolone resistance in clinical isolates of Salmonella Typhi from India.

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    Manoj Kumar

    Full Text Available Enteric fever is a major cause of morbidity in several parts of the Indian subcontinent. The treatment for typhoid fever majorly includes the fluoroquinolone group of antibiotics. Excessive and indiscriminate use of these antibiotics has led to development of acquired resistance in the causative organism Salmonella Typhi. The resistance towards fluoroquinolones is associated with mutations in the target gene of DNA Gyrase. We have estimated the Minimum Inhibitory Concentration (MIC of commonly used fluoroquinolone representatives from three generations, ciprofloxacin, ofloxacin, levofloxacin and moxifloxacin, for 100 clinical isolates of Salmonella Typhi from patients in the Indian subcontinent. The MICs have been found to be in the range of 0.032 to 8 μg/ml. The gene encoding DNA Gyrase was subsequently sequenced and point mutations were observed in DNA Gyrase in the quinolone resistance determining region comprising Ser83Phe/Tyr and Asp87Tyr/Gly. The binding ability of these four fluoroquinolones in the quinolone binding pocket of wild type as well as mutant DNA Gyrase was computationally analyzed by molecular docking to assess their differential binding behaviour. This study has revealed that mutations in DNA Gyrase alter the characteristics of the binding pocket resulting in the loss of crucial molecular interactions and consequently decrease the binding affinity of fluoroquinolones with the target protein. The present study assists in understanding the underlying molecular and structural mechanism for decreased fluoroquinolone susceptibility in clinical isolates as a consequence of mutations in DNA Gyrase.

  16. Serovars of Salmonella isolated from Danish turkeys between 1995 and 2000 and their antimicrobial resistance.

    Science.gov (United States)

    Pedersen, K; Hansen, H C; Jørgensen, J C; Borck, B

    2002-04-13

    The prevalence of Salmonella serovars and their antimicrobial resistance patterns were investigated among Danish turkeys between 1995 and 2000, by sampling the flocks approximately 14 days before they were slaughtered. Within the flocks, the prevalence of salmonella varied from 7.1 per cent to 25 per cent, and 24 different serovars were detected. The five most prevalent, which accounted for 58.5 per cent of the isolates were Salmonella Heidelberg (16.2 per cent of the isolates), Salmonella Agona (15.8 per cent), Salmonella Derby (12.4 per cent), Salmonella Muenster (7.3 per cent) and Salmonella Anatum (6.8 per cent). In addition, a few rough isolates and isolates belonging to the antigenically incomplete formulae 6,7:-:- and 4,12:b:- were found. The level of antimicrobial resistance was low; the highest resistance was recorded to ampicillin (13.7 per cent) and streptomycin (9.0 per cent) followed by tetracycline (8.5 per cent), sulphonamides (7.7 per cent) and spectinomycin (4.7 per cent). Resistance to quinolones was very low: four isolates were resistant to nalidixic acid, and only one was resistant to enrofloxacin. No resistance was recorded to colistin, apramycin, ceftiofur, florfenicol, or amoxycillin with clavulanic acid. Only 24 isolates were resistant to two or more compounds in various combinations of up to six compounds; one Salmonella Havana isolate was resistant to six compounds. Six isolates were serovar Typhimurium, but none of them belonged to phage type DT104.

  17. Membrane permeability, a pivotal function involved in antibiotic resistance and virulence in Enterobacter aerogenes clinical isolates.

    Science.gov (United States)

    Lavigne, J-P; Sotto, A; Nicolas-Chanoine, M-H; Bouziges, N; Bourg, G; Davin-Regli, A; Pagès, J-M

    2012-06-01

    Imipenem-susceptible E. aerogenes isolates exhibiting extended spectrum β-lactamases, target mutations and a basal efflux expression, were identified in five patients. After imipenem treatment, imipenem-intermediate susceptible (IMI-I) or resistant (IMI-R) isolates emerged in these patients. Alteration in porin synthesis and increase in efflux expression were observed in the IMI-I isolates whereas complete loss of the porins, LPS alteration and efflux overexpression were observed in the IMI-R isolates. Bacterial virulence of the strains was investigated by the Caenorhabditis elegans model. The IMI-R isolates were shown to be significantly less virulent than the IMI-susceptible or IMI-I isolates. The pleiotropic membrane alteration and its associated fitness burden exhibited by E. aerogenes isolates influence their antibiotic resistance and their virulence behaviour. These findings highlight the balance between the low permeability-related resistance and virulence and their relationships with the treatment of resistant pathogens.

  18. Genotypic characterization of quinolone resistant-Escherichia coli isolates from retail food in Morocco.

    Science.gov (United States)

    Nayme, Kaotar; Barguigua, Abouddihaj; Bouchrif, Brahim; Karraouan, Bouchra; El Otmani, Fatima; Elmdaghri, Naima; Zerouali, Khalid; Timinouni, Mohammed

    2017-02-01

    This study was conducted to assess the retail food as a possible vehicle for antimicrobial resistant, particularly quinolones resistant and pathogenic Escherichia coli. We determined the prevalence and characteristics of nalidixic acid (Nal) resistant E. coli isolates from diverse retail food samples. In all, 70 (28%) of 250 E. coli isolates studied were Nal-resistant E. coli and 91% of these were multi-drug resistant. Plasmid mediated quinolone resistance genes were identified in 32 isolates, including aac(6')-Ib-cr (n = 16), qnrS1 (n = 11) and qnrB19 (n = 7). Mutations in gyr A and par C genes were detected among 80% of the isolates, and the isolates showed substitution Ser83-Leu and Asp87-Asn in gyrA and Ser80-Ile in parC. In addition, three different gene cassettes were identified (aadA1, aadA7, aac(3)-Id) in 18%. Virulence-associated genes stx1, eae, sfa, hlyA and stx2 were found in six (8%), three (4%), two (3%), three (4%) and three (4%) isolates, respectively. E. coli isolates of phylogenetic group A were dominant (64%, 45/70). Pulsed field gel electrophoresis revealed none epidemiological relationship between these isolates. The results of this work report the higher frequency of Nal-resistant E. coli isolates from Moroccan retail food samples including MDR and pathogenic isolates.

  19. Analysis of embCAB mutations associated with ethambutol resistance in multidrug-resistant mycobacterium tuberculosis isolates from China.

    Science.gov (United States)

    Zhao, Li-Li; Sun, Qing; Liu, Hai-Can; Wu, Xiao-Cui; Xiao, Tong-Yang; Zhao, Xiu-Qin; Li, Gui-Lian; Jiang, Yi; Zeng, Chun-Yan; Wan, Kang-Lin

    2015-04-01

    Ethambutol (EMB) plays a pivotal role in the chemotherapy of drug-resistant tuberculosis (TB), including multidrug-resistant tuberculosis (MDR-TB). Resistance to EMB is considered to be caused by mutations in the embCAB operon (embC, embA, and embB). In this study, we analyzed the embCAB mutations among 139 MDR-TB isolates from China and found a possible association between embCAB operon mutation and EMB resistance. Our data indicate that 56.8% of MDR-TB isolates are resistant to EMB, and 82.2% of EMB-resistant isolates belong to the Beijing family. Overall, 110 (79.1%) MDR-TB isolates had at least one mutation in the embCAB operon. The majority of mutations were present in the embB gene and the embA upstream region, which also displayed significant correlations with EMB resistance. The most common mutations occurred at codon 306 in embB (embB306), followed by embB406, embA(-16), and embB497. Mutations at embB306 were associated with EMB resistance. DNA sequencing of embB306-497 was the best strategy for detecting EMB resistance, with 89.9% sensitivity, 58.3% specificity, and 76.3% accuracy. Additionally, embB306 had limited value as a candidate predictor for EMB resistance among MDR-TB infections in China.

  20. Deletions in a ribosomal protein-coding gene are associated with tigecycline resistance in Enterococcus faecium.

    Science.gov (United States)

    Niebel, Marc; Quick, Joshua; Prieto, Ana Maria Guzman; Hill, Robert L R; Pike, Rachel; Huber, Damon; David, Miruna; Hornsey, Michael; Wareham, David; Oppenheim, Beryl; Woodford, Neil; van Schaik, Willem; Loman, Nicholas

    2015-11-01

    Enterococcus faecium is an emerging nosocomial pathogen associated with antibiotic therapy in the hospital environment. Whole-genome sequences were determined for three pairs of related, consecutively collected E. faecium clinical isolates to determine putative mechanisms of resistance to tigecycline. The first isolates (1S, 2S and 3S) in each of the three pairs were sensitive to tigecycline [minimum inhibitory concentration (MIC) of 0.125 mg/L]. Following tigecycline therapy, the second isolate in each pair demonstrated increased resistance to tigecycline. Two isolates (1R and 2R) were resistant (MIC of 8 mg/L) and one isolate (3I) demonstrated reduced susceptibility (MIC of 0.5 mg/L). Mutations distinguishing each pair of sensitive and resistant isolates were determined through alignment to a reference genome and variant detection. In addition, a de novo assembly of each isolate genome was constructed to confirm mutations. A total of 16 mutations in eleven coding sequences were determined. Mutations in the rpsJ gene, which encodes a structural protein forming part of the 30S ribosomal subunit, were detected in each of the pairs. Mutations were in regions proximal to the predicted tigecycline-binding site. Predicted amino acid substitutions were detected in 1R and 3I. The resistant strains were additionally associated with deletions of 15 nucleotides (2R) and 3 nucleotides (1R). This study confirms that amino acid substitutions in rpsJ contribute towards reduced susceptibility to tigecycline and suggests that deletions may be required for tigecycline resistance in E. faecium.

  1. First antimicrobial resistance data and genetic characteristics of Neisseria gonorrhoeae isolates from Estonia, 2009–2013

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    D. Golparian

    2014-09-01

    Full Text Available Gonorrhoea is a sexually transmitted infection with major public health implications and Neisseria gonorrhoeae has developed resistance to all antimicrobials introduced for treatment. Enhanced surveillance of antimicrobial resistance in N. gonorrhoeae is crucial globally. This is the first internationally reported antimicrobial resistance data for N. gonorrhoeae from Estonia (44 isolates cultured in 2009–2013. A high prevalence of resistance was observed for azithromycin, ciprofloxacin and tetracycline. One and two isolates with resistance and decreased susceptibility to the last remaining first-line treatment option ceftriaxone, respectively, were identified. It is crucial to implement surveillance of gonococcal antimicrobial resistance (ideally also treatment failures in Estonia.

  2. Carbapenem-resistance and pathogenicity of bovine Acinetobacter indicus-like isolates

    Science.gov (United States)

    Leidner, Ursula; Semmler, Torsten; Scheufen, Sandra; Ewers, Christa

    2017-01-01

    The objective of this study was to characterize blaOXA-23 harbouring Acinetobacter indicus-like strains from cattle including genomic and phylogenetic analyses, antimicrobial susceptibility testing and evaluation of pathogenicity in vitro and in vivo. Nasal and rectal swabs (n = 45) from cattle in Germany were screened for carbapenem-non-susceptible Acinetobacter spp. Thereby, two carbapenem resistant Acinetobacter spp. from the nasal cavities of two calves could be isolated. MALDI-TOF mass spectrometry and 16S rDNA sequencing identified these isolates as A. indicus-like. A phylogenetic tree based on partial rpoB sequences indicated closest relation of the two bovine isolates to the A. indicus type strain A648T and human clinical A. indicus isolates, while whole genome comparison revealed considerable intraspecies diversity. High mimimum inhibitory concentrations were observed for carbapenems and other antibiotics including fluoroquinolones and gentamicin. Whole genome sequencing and PCR mapping revealed that both isolates harboured blaOXA-23 localized on the chromosome and surrounded by interrupted Tn2008 transposon structures. Since the pathogenic potential of A. indicus is unknown, pathogenicity was assessed employing the Galleria (G.) mellonella infection model and an in vitro cytotoxicity assay using A549 human lung epithelial cells. Pathogenicity in vivo (G. mellonella killing assay) and in vitro (cytotoxicity assay) of the two A. indicus-like isolates was lower compared to A. baumannii ATCC 17978 and similar to A. lwoffii ATCC 15309. The reduced pathogenicity of A. indicus compared to A. baumannii correlated with the absence of important virulence genes encoding like phospholipase C1+C2, acinetobactin outer membrane protein BauA, RND-type efflux system proteins AdeRS and AdeAB or the trimeric autotransporter adhesin Ata. The emergence of carbapenem-resistant A. indicus-like strains from cattle carrying blaOXA-23 on transposable elements and revealing genetic

  3. Aqueous fractionation yields chemically stable lupin protein isolates

    NARCIS (Netherlands)

    Berghout, J.A.M.; Marmolejo-Garcia, C.; Berton-Carabin, C.C.; Nikiforidis, C.V.; Boom, R.M.; Goot, van der A.J.

    2015-01-01

    The chemical stability of lupin protein isolates (LPIs) obtained through aqueous fractionation (AF, i.e. fractionation without the use of an organic solvent) at 4 °C or 20 °C was assessed. AF of lupin seeds results in LPIs containing 2 wt.% oil. This oil is composed of mono- and poly-unsaturated fat

  4. Occurrence of antibiotic resistance and characterization of resistant genes and integrons in Enterobacteriaceae isolated from integrated fish farms south China

    Science.gov (United States)

    Su, Hao-Chang; Ying, Guang-Guo; Tao, Ran; Zhang, Rui-Quan; Fogarty, Lisa R.; Kolpin, Dana W.

    2011-01-01

    Antibiotics are still widely applied in animal husbandry to prevent diseases and used as feed additives to promote animal growth. This could result in antibiotic resistance to bacteria and antibiotic residues in animals. In this paper, Enterobacteriaceae isolated from four integrated fish farms in Zhongshan, South China were tested for antibiotic resistance, tetracycline resistance genes, sulfonamide resistance genes, and class 1 integrons. The Kirby-Bauer disk diffusion method and polymerase chain reaction (PCR) assays were carried out to test antibiotic susceptibility and resistance genes, respectively. Relatively high antibiotic resistance frequencies were found, especially for ampicillin (80%), tetracycline (52%), and trimethoprim (50%). Out of 203 Enterobacteriaceae isolates, 98.5% were resistant to one or more antibiotics tested. Multiple antibiotic resistance (MAR) was found highest in animal manures with a MAR index of 0.56. Tetracycline resistance genes (tet(A), tet(C)) and sulfonamide resistance genes (sul2) were detected in more than 50% of the isolates. The intI1 gene was found in 170 isolates (83.7%). Both classic and non-classic class 1 integrons were found. Four genes, aadA5, aadA22, dfr2, and dfrA17, were detected. To our knowledge, this is the first report for molecular characterization of antibiotic resistance genes in Enterobacteriaceae isolated from integrated fish farms in China and the first time that gene cassette array dfrA17-aadA5 has been detected in such fish farms. Results of this study indicated that fish farms may be a reservoir of highly diverse and abundant antibiotic resistant genes and gene cassettes. Integrons may play a key role in multiple antibiotic resistances posing potential health risks to the general public and aquaculture.

  5. Enterococcus faecalis as multidrug resistance strains in clinical isolates in Imam Reza Hospital in Kermanshah, Iran.

    Science.gov (United States)

    Mohammadi, F; Ghafourian, S; Mohebi, R; Taherikalani, M; Pakzad, I; Valadbeigi, H; Hatami, V; Sadeghifard, N

    2015-01-01

    The current study aimed to investigate the prevalence of vancomycin-resistant Enterococcus in E. faecalis and E. faecium and antimicrobial susceptibility patterns, then dominant genes responsible for vancomycin resistance were determined. For this propose, 180 clinical isolates of Enterococcus were subjected for identification and antibiotic susceptibility assay. Then, the gene responsible vancomycin resistant strains were determined. The results demonstrated the E. faecalis as a dominant Enterococcus. Resistance to erythromycin was dominant and multidrug resistance strains observed in E. faecalis. vanA was responsible for vancomycin resistance. In conclusion, a high rate of resistance to antibiotics in Enterococcus is clearly problematic, and a novel strategy is needed to decrease resistance in Enterococcus.

  6. Multidrug and vancomycin resistance among clinical isolates of ...

    African Journals Online (AJOL)

    2017-09-03

    Sep 3, 2017 ... Keywords: Bacterial resistance, vancomycin resistant S. aureus, susceptibility studies, agar dilution. ..... therapy, a lack of sufficient knowledge on the danger of ... lin-resistant Staphylococcus aureus as a public-health threat.

  7. Antimicrobial Resistance and Resistance Genes in Aerobic Bacteria Isolated from Pork at Slaughter.

    Science.gov (United States)

    Li, Lili; Heidemann Olsen, Rikke; Ye, Lei; Yan, He; Nie, Qing; Meng, Hecheng; Shi, Lei

    2016-04-01

    The aim of this study was to investigate the phenotypic and genotypic antimicrobial resistance, integrons, and transferability of resistance markers in 243 aerobic bacteria recovered from pork at slaughter in the People's Republic of China. The organisms belonged to 22 genera of gram-negative bacteria (92.2%) and gram-positive bacteria (7.8%). High levels of resistance were detected to tetracycline, trimethoprim-sulfamethoxazole, and ampicillin (36.2 to 54.3%), and lower levels were detected to nitrofurantoin, cefotaxime, gentamicin, ciprofloxacin, and chloramphenicol (7.8 to 29.2%). Across species, genes conferring antimicrobial resistance were observed with the following frequencies: blaTEM, 40.7%; blaCMY-2, 15.2%; blaCTX-M, 11.5%; sul2, 27.2%; sul1, 14.4%; tet(A), 5.4%; tet(L), 5.4%; tet(M), 5.0%; tet(E), 3.7%; tet(C), 3.3%; tet(S), 2.5%; and tet(K), 0.8%. Various antimicrobial resistance genes were found in new carriers: blaTEM in Lactococcus garvieae, Myroides odoratimimus, Aeromonas hydrophila, Staphylococcus sciuri, Raoultella terrigena, Macrococcus caseolyticus, Acinetobacter ursingii, Sphingobacterium sp., and Oceanobacillus sp.; blaCMY-2 in Lactococcus lactis, Klebsiella oxytoca, Serratia marcescens, Acinetobacter baumannii, and Myroides phaeus; tet(L) in M. caseolyticus; sul1 in Vibrio cincinnatiensis; sul2 in Acinetobacter bereziniae, Acinetobacter johnsonii, and V. cincinnatiensis; and the class 1 integron and gene cassette aadA2 in V. cincinnatiensis. Approximately 6.6% of isolates contained class 1 integrons, and one isolate harbored class 2 integrons. Plasmid associated intI1 and androgen receptor- encoding genes were transferred into Escherichia coli J53 and E. coli DH5α by conjugation and transformation experiments, respectively. Our study highlights the importance of aerobic bacteria from pork as reservoirs for antimicrobial resistance genes and mobile genetic elements that can readily be transferred intra- and interspecies.

  8. Molecular serotyping and antimicrobial resistance profiles of Actinobacillus pleuropneumoniae isolated from pigs in South Korea.

    Science.gov (United States)

    Kim, Boram; Hur, Jin; Lee, Ji Yeong; Choi, Yoonyoung; Lee, John Hwa

    2016-09-01

    Actinobacillus pleuropneumoniae (APP) causes porcine pleuropneumonia (PP). Serotypes and antimicrobial resistance patterns in APP isolates from pigs in Korea were examined. Sixty-five APP isolates were genetically serotyped using standard and multiplex PCR (polymerase chain reaction). Antimicrobial susceptibilities were tested using the standardized disk-agar method. PCR was used to detect β-lactam, gentamicin and tetracycline-resistance genes. The random amplified polymorphic DNA (RAPD) patterns were determined by PCR. Korean pigs predominantly carried APP serotypes 1 and 5. Among 65 isolates, one isolate was sensitive to all 12 antimicrobials tested in this study. Sixty-two isolates was resistant to tetracycline and 53 isolates carried one or five genes including tet(B), tet(A), tet(H), tet(M)/tet(O), tet(C), tet(G) and/or tet(L)-1 markers. Among 64 strains, 9% and 26.6% were resistance to 10 and three or more antimicrobials, respectively. Thirteen different antimicrobial resistance patterns were observed and RAPD analysis revealed a separation of the isolates into two clusters: cluster II (6 strains resistant to 10 antimicrobials) and cluster I (the other 59 strains). Results show that APP serotypes 1 and 5 are the most common in Korea, and multi-drug resistant strains are prevalent. RAPD analysis demonstrated that six isolates resistant to 10 antimicrobials belonged to the same cluster.

  9. Effect of Alternaria solani exudates on resistant and susceptible potato cultivars from two different pathogen isolates

    Science.gov (United States)

    The resistance phenotypes of two potato cultivars to two isolates of Alternaria solani, causal agent of early blight, were studied under greenhouse conditions. The two isolates contain varying degrees of aggressiveness on both susceptible and resistant phenotypes of potatoes. A bioassay was used to ...

  10. Clonal origin of aminoglycoside-resistant Citrobacter freundii isolates in a Danish county

    DEFF Research Database (Denmark)

    Norskov-Lauritsen, N.; Sandvang, Dorthe; Hedegaard, J.;

    2001-01-01

    During 1997, attention was drawn to an increased frequency of aminoglycoside-resistant Citrobacter freundii in a Danish county, when a total of 24 resistant C. freundii isolates was detected. In this study, 15 such isolates were typed by pulsed-field gel electrophoresis, riboprinting and partial...

  11. Draft Genome of the Multidrug-Resistant Acinetobacter baumannii Strain A155 Clinical Isolate.

    Science.gov (United States)

    Arivett, Brock A; Fiester, Steven E; Ream, David C; Centrón, Daniela; Ramírez, Maria S; Tolmasky, Marcelo E; Actis, Luis A

    2015-03-26

    Acinetobacter baumannii is a bacterial pathogen with serious implications on human health, due to increasing reports of multidrug-resistant strains isolated from patients. Total DNA from the multidrug-resistant A. baumannii strain A155 clinical isolate was sequenced to greater than 65× coverage, providing high-quality contig assemblies.

  12. Characterization of erythromycin-resistant Streptococcus pneumoniae isolates causing invasive diseases in Chinese children

    Institute of Scientific and Technical Information of China (English)

    MA Xiang; YAO Kai-hu; XIE Gui-lin; ZHENG YUE-jie; WANG Chuan-qing; SHANG Yun-xiao; WANG Hui-yun

    2013-01-01

    Background Erythromycin-resistant Streptococcus pneumoniae isolates that causing invasive pneumococcal diseases (IPD) in Chinese children remain uncharacterized.This study aims to identify the resistance genes associated with erythromycin resistance and to determine the genetic relationships of IPD isolates in Chinese children.Methods A total of 171 S.pneumoniae strains were isolated from 11 medical centers in China from 2006 to 2008.All the isolates were characterized via serotyping and antibiotic susceptibility determination.The erythromycin-resistant isolates were further characterized via ermB and mefA gene detection,multi-locus sequence typing analysis,and pulsed-field gel electrophoresis.Results A total of 164 (95.9%) isolates showed resistance to erythromycin,of which 162 strains with high high-level resistance (MIC ≥ 256 μg/ml).A total of 104 (63.4%) isolates carry the ermB gene alone,whereas 59 (36.0%) harbor both ermB and mefA genes.Of the 59 strains,54 were of serotypes 19A and 19F and were identified as highly clonal and related to the Taiwan19F-14 clone.Conclusions The erythromycin resistance rate in IPD isolates is significantly high and is predominantly mediated by the ermB gene.Isolates that carry both ermB and mefA genes are predominantly of serotypes 19A and 19F.

  13. MLST Genotypes and Antibiotic Resistance of Campylobacter spp. Isolated from Poultry in Grenada

    Science.gov (United States)

    Stone, Diana; Davis, Margaret; Baker, Katherine; Besser, Tom; Roopnarine, Rohini; Sharma, Ravindra

    2013-01-01

    This study determined whether multilocus sequence types (MLST) of Campylobacter from poultry in 2 farms in Grenada, West Indies, differed by farm, antimicrobial resistance and farm antibiotic use. Farm A used fluoroquinolones in the water and Farm B used tetracyclines. The E-test was used to determine resistance of isolates to seven antibiotics. PCR of the IpxA gene confirmed species and MLST was used to characterize 38 isolates. All isolates were either C. jejuni or C. coli. Farm antibiotic use directly correlated with antimicrobial resistance of Campylobacter isolates. Almost 80% of the isolates from Farm A were fluoroquinolone resistant and 17.9% of the isolates from Farm B were fluoroquinolone resistant. All Campylobacter isolates from Farm A were tetracycline sensitive, whereas 35.7% of isolates from Farm B were tetracycline resistant. Six previously recognized sequence types (STs) and 2 novel STs were identified. Previously recognized STs were those overwhelmingly reported from poultry and humans globally. Isolates with the same ST did not always have the same antibiotic resistance profile. There was little ST overlap between the farms suggesting that within-farm transmission of Campylobacter genotypes may dominate. MLST typing was useful for tracking Campylobacter spp. among poultry units and can help elucidate Campylobacter host-species population structure and its relevance to human health. PMID:23555097

  14. MLST Genotypes and Antibiotic Resistance of Campylobacter spp. Isolated from Poultry in Grenada

    Directory of Open Access Journals (Sweden)

    Diana Stone

    2013-01-01

    Full Text Available This study determined whether multilocus sequence types (MLST of Campylobacter from poultry in 2 farms in Grenada, West Indies, differed by farm, antimicrobial resistance and farm antibiotic use. Farm A used fluoroquinolones in the water and Farm B used tetracyclines. The E-test was used to determine resistance of isolates to seven antibiotics. PCR of the IpxA gene confirmed species and MLST was used to characterize 38 isolates. All isolates were either C. jejuni or C. coli. Farm antibiotic use directly correlated with antimicrobial resistance of Campylobacter isolates. Almost 80% of the isolates from Farm A were fluoroquinolone resistant and 17.9% of the isolates from Farm B were fluoroquinolone resistant. All Campylobacter isolates from Farm A were tetracycline sensitive, whereas 35.7% of isolates from Farm B were tetracycline resistant. Six previously recognized sequence types (STs and 2 novel STs were identified. Previously recognized STs were those overwhelmingly reported from poultry and humans globally. Isolates with the same ST did not always have the same antibiotic resistance profile. There was little ST overlap between the farms suggesting that within-farm transmission of Campylobacter genotypes may dominate. MLST typing was useful for tracking Campylobacter spp. among poultry units and can help elucidate Campylobacter host-species population structure and its relevance to human health.

  15. Antibiotic Resistance Pattern and Biofilm Formation Ability of Clinically Isolates of Salmonella enterica Serotype typhimurium

    Directory of Open Access Journals (Sweden)

    Hadi Ghasemmahdi

    2015-05-01

    Full Text Available Background: The emergence of antimicrobial-resistant bacteria with biofilm formation ability may be a major threat to public health and food safety and sanitation. Objectives: The aim of this study was to determine antibiotic resistance patterns and biofilm production characteristics of Salmonella typhimurium isolated from different species of birds. Materials and Methods: The antibiotic resistance patterns of 38 pre-identified isolates were screened by standard Kirby-Bauer disc-diffusion method performed on Mueller–Hinton agar to a panel of 17 antibiotics. The extent of biofilm formation was measured by Microtiter plate (MTP-based systems. Results: The highest antimicrobial resistance was detected against nalidixic acid (97%, followed by doxycycline (86%, colistin (84%, streptomycin (84% and tetracycline (84%. All isolates were sensitive to amikacin (100% and 97% and 95% of the isolates were sensitive to ceftazidime and ceftriaxone, respectively. Twenty one different antibiotic resistance patterns were observed among S. typhimurium isolates. According to the results of the microtitre plate biofilm assay, there was a wide variation in biofilm forming ability among S. typhimurium isolates. Most of the isolates (60.52% were not capable of producing biofilm, while 26.31%, 7.89%, and 5.26% isolates were weak, strong and moderate biofilm producers, respectively. Conclusions: It was concluded that nearly all S. typhimurium isolates revealed a high multiple antibiotic resistant with low biofilm forming capabilities which proposed low association between biofilm formation and antibiotic resistance of a major food important pathogen.

  16. Antimicrobial resistance in equine faecal Escherichia coli isolates from North West England

    Directory of Open Access Journals (Sweden)

    Williams Nicola J

    2010-04-01

    Full Text Available Abstract Background Escherichia coli isolates of equine faecal origin were investigated for antibiotic resistance, resistance genes and their ability to perform horizontal transfer. Methods In total, 264 faecal samples were collected from 138 horses in hospital and community livery premises in northwest England, yielding 296 resistant E. coli isolates. Isolates were tested for susceptibility to antimicrobial drugs by disc diffusion and agar dilution methods in order to determine minimum inhibitory concentrations (MIC. PCR amplification was used to detect genes conferring resistance to: ampicillin (TEM and SHV beta-lactamase, chloramphenicol (catI, catII, catIII and cml, tetracycline (tetA, tetB, tetC, tetD, tet E and tetG, and trimethoprim (dfrA1, dfrA9, dfrA12, dfrA13, dfr7, and dfr17. Results The proportion of antibiotic resistant isolates, and multidrug resistant isolates (MDR was significantly higher in hospital samples compared to livery samples (MDR: 48% of hospital isolates; 12% of livery isolates, p dfr, TEM beta-lactamase, tet and cat, conferring resistance to trimethoprim, ampicillin, tetracycline and chloramphenicol, respectively. Within each antimicrobial resistance group, these genes occurred at frequencies of 93% (260/279, 91%, 86.8% and 73.5%, respectively; with 115/296 (38.8% found to be MDR isolates. Conjugation experiments were performed on selected isolates and MDR phenotypes were readily transferred. Conclusions Our findings demonstrate that E. coli of equine faecal origin are commonly resistant to antibiotics used in human and veterinary medicine. Furthermore, our results suggest that most antibiotic resistance observed in equine E. coli is encoded by well-known and well-characterized resistant genes common to E. coli from man and domestic animals. These data support the ongoing concern about antimicrobial resistance, MDR, antimicrobial use in veterinary medicine and the zoonotic risk that horses could potentially pose to

  17. The importance of inducible clindamycin resistance in enterotoxin positive S. aureus isolated from clinical samples

    Directory of Open Access Journals (Sweden)

    Memariani M

    2009-07-01

    Full Text Available "nBackground: Clindamycin is a suitable antibiotic for treatment of skin and soft tissue infections. Moreover, it can suppress toxin production in many pathogenic bacteria such as S. aureus. There are two mechanisms of resistance in this antibiotic. Constitutive resistance can be detected by standard disk diffusion method but in the case of inducible resistance, D-test should be carried out. The main aim of this study is to determine prevalence of clindamycin inducible resistance among methicillin resistant and susceptible isolates of S. aureus isolated from different clinical samples. "nMethods: A total of 87 clinical isolates from clinical samples were collected. Methicillin resistance was determined using standard disk diffusion method. Subsequently, D-test was carried out according to CLSI guideline. Presence of the sea gene (enterotoxin A was detected by PCR using specific primers. "nResults: Out of 87 isolates, 18(20.7% were clindamycin inducible resistant while constitutive resistance was detected among 21(24.1% isolates. The 95% Confidence intervals for the proportion of inducible clindamycin resistance among clinical isolates of S. aureus was 12.2% to 29.2%. The inducible phenotype in MRSA isolates was more common than that of MSSA isolates (33.3% vs 5.1%.Significant differences were found between prevalence of inducible clindamycin resistance and type of infection (p=0.045. Importantly, there was a significant correlation between sea gene and the constitutive/inducible resistance (p<0.0001. "nConclusions: Due to the high prevalence of clindamycin inducible resistance among clinical isolates of S. aureus, we recommend D-test to avoid treatment failure.

  18. Resistance to Sulfonamides and Dissemination of sul Genes Among Salmonella spp. Isolated from Food in Poland.

    Science.gov (United States)

    Mąka, Łukasz; Maćkiw, Elżbieta; Ścieżyńska, Halina; Modzelewska, Magdalena; Popowska, Magdalena

    2015-05-01

    Antimicrobial resistance of pathogenic bacteria, including Salmonella spp., is an emerging problem of food safety. Antimicrobial use can result in selection of resistant organisms. The food chain is considered a route of transmission of resistant pathogens to humans. In many European countries, sulfonamides are one of the most commonly used antimicrobials. The aim of our investigation was to assess the prevalence of sul genes and plasmid occurrence among sulfonamide-resistant Salmonella spp. Eighty-four sulfonamide-resistant isolates were collected in 2008 and 2013 from retail products in Poland. Minimal inhibitory concentration of all of these isolates was ≥1024 μg/mL. Resistant isolates were tested for the presence of sul1, sul2, sul3, and int1 genes by using multiplex polymerase chain reaction. In total, 44.0% (37/84) isolates carried the sul1 gene, 46.4% (39/84) were sul2 positive, while the sul3 gene was not detected in any of the sulfonamide-resistant isolates tested. It was found that 3.6% (3/84) of resistant Salmonella spp. contained sul1, sul2, and intI genes. All 33 intI-positive isolates carried the sul1 gene. Eleven of the sulfonamide-resistant isolates were negative for all the sul genes. Most of the sulfonamide-resistant Salmonella spp. harbored plasmids; only in eight isolates were no plasmids detected. Generally, the size of the plasmids ranged from approximately 2 kb to ≥90 kb. Our results revealed a relatively a high prevalence of sulfonamides-resistant Salmonella spp. isolated from retail food. Additionally, we have detected a high dissemination of plasmids and class 1 integrons that may enhance the spread of resistance genes in the food chain.

  19. Nature of protein-protein interactions during the gelation of canola protein isolate networks.

    Science.gov (United States)

    Kim, Jae He; Varankovich, Natallia V; Stone, Andrea K; Nickerson, Michael T

    2016-11-01

    The nature of interactions involved during the gelation of a canola protein isolate was investigated using rheology and fractal imaging at neutral pH as a function of protein concentration (5.0-9.0% w/w). The onset of denaturation and the denaturation temperature by differential scanning calorimetry for canola protein isolate (CPI; 98.2% protein) was 78.6°C and 87.1°C, respectively. Rheological testing determined the gelation temperature (Tgel) to be ~87-90°C for all concentrations. The log % strain at break increased from 1.70 to 1.80 as CPI concentration increased from 5.0 to 7.0% (w/w). Rheological testing of CPI in the presence of destabilizing agents, NaCl (0.1 and 0.5M), urea (0.1, 0.5, 1 and 5M) and 2-β-mercaptoethanol (0.1 and 2%), was performed. Samples with NaCl and urea (0.1-1M) had similar temperature profiles and Tgel values to CPI alone whereas no gel was formed with the addition of 5M urea and 2-β-mercaptoethanol reduced the strength of the gel network. Fractal dimension and lacunarity was analyzed using CLSM imaging. The fractal dimension value for all CPI concentrations was ~1.5. The lacunarity of the gel decreased from 0.62 to 0.41 as the concentration of CPI increased from 5 to 7% (w/w). Mechanistic understanding of CPI aggregation and network formation will enable the food industry to better tailor food structure when CPI is present as ingredient. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Transmissible antibiotic resistance in Salmonella isolated from random-source cats purchased for use in research.

    Science.gov (United States)

    Beaucage, C M; Fox, J G

    1979-06-01

    Salmonella isolates from random-source cats designated for use in research were examined for antibiotic susceptibilities and the presence of plasmids containing R factors. The serotypes studied were Salmonella derby, S typhimurium, S anatum, S enteritidis, and S bredeney. Eighty percent of the isolates were resistant to one or more antibiotics. The greatest frequency of resistance was to streptomycin. The majority of the salmonella isolates transferred all or a part of their antibiotic resistance to an Escherichia coli K-12 recipient. Thermosensitive R factors were found in two S typhimurium isolates.

  1. ERG11 mutations associated with azole resistance in Candida albicans isolates from vulvovaginal candidosis patients

    Institute of Scientific and Technical Information of China (English)

    Bin Wang; Li-Hua Huang; Ji-Xue Zhao; Man Wei; Hua Fang; Dan-Yang Wang; Hong-Fa Wang; Ji-Gang Yin; Mei Xiang

    2015-01-01

    Objective:To investigate the azole susceptibility of Candida albicans (C. albicans) from vulvovaginal candidosis patients and to analyze the relationship between ERG11 gene mutations in these isolates and azole resistance. Methods:Three hundred and two clinical isolates of Candida species were collected. Azole susceptibility was tested in vitro in microdilution studies. The ERG11 genes of 17 isolates of C. albicans (2 susceptibles, 5 dose-dependent resistants and 10 resistants) were amplified and sequenced. Results:Of the 302 isolates collected, 70.2%were C. albicans, of which 8.5%, 3.8%and 4.2%were resistant to fluconazole, itraconazole and voriconazole, respectively. In total, 27 missense mutations were detected in ERG11 genes from resistant/susceptible dose-dependent isolates. Among them, Y132H, A114S, and Y257H substitutions were most prevalent and were known to cause fluconazole resistance. G464S and F72S also has been proved to cause fluconazole resistance. Two novel substitutions (T285A, S457P) in hotspot regions were identified. Conclusions:Twenty seven mutations in the ERG11 gene were identified in azole-resistant C. albicans isolates, which indicated a possible relation with the increase in resistance to azole drugs and the recurrence of vulvovaginal candidosis. The relationship of two novel substitutions (T285A, S457P) with fluconazole resistance needs to be further verified by site-directed mutagenesis.

  2. Assessment of antibiotic resistance phenotype and integrons in Salmonella enterica serovar Typhimurium isolated from swine.

    Science.gov (United States)

    Rayamajhi, Nabin; Kang, Sang Gyun; Kang, Mi Lan; Lee, Hee Soo; Park, Kyung Yoon; Yoo, Han Sang

    2008-10-01

    Salmonella enterica serovar Typhimurium (S. Typhimurium) isolated and identified from swine were subjected for the analysis of antibiotic resistance pattern and clinically important class 1 and 2 integrons. In addition, S. Typhimurium isolates exhibiting ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, tetracycline and florfenicol (ACSSuTF) resistance pattern as described in most Salmonella enterica serotype Typhimurium definitive type 104 (DT104) were characterized by polymerase chain reaction. All the isolates were resistant to more than four antibiotics and showed the highest resistance to streptomycin (94.1%), followed by tetracycline (90.1%), ampicillin (64.7%), chloramphenicol (56.8%) and gentamicin (54.9%). MIC value for the ten isolates ranged between 0.125-2 mug/ml for ciprofloxacin. Among the beta-lactams used, only one of the isolate exhibited resistance to ceftiofur (MIC 8 microg/ml). Sixty eight percent of these multi drug resistance (MDR) S. Typhimurium isolates carried clinically important class 1 integron with 1kb (aadA) and/or 2kb (dhfrXII-orfF-aadA2) resistance gene cassettes. This study reports the increasing trend of multi drug resistance (MDR) S. Typhimurium with clinically important class 1 integron in pigs. In addition, emergence of the ACSSuTF-type resistance in S. Typhimurium PT other than DT104 may limit the use of resistance gene markers in its detection methods by PCR.

  3. Antimicrobial Resistance in Enterococcus spp. Isolated from Environmental Samples in an Area of Intensive Poultry Production

    Directory of Open Access Journals (Sweden)

    Patricia A. Chambers

    2013-03-01

    Full Text Available Enterococcus spp. from two poultry farms and proximate surface and ground water sites in an area of intensive poultry production were tested for resistance to 16 clinical antibiotics. Resistance patterns were compared to assess trends and possible correlations for specific antimicrobials and levels of resistance. Enterococci were detected at all 12 surface water sites and three of 28 ground water sites. Resistance to lincomycin, tetracycline, penicillin and ciprofloxacin in poultry litter isolates was high (80.3%, 65.3%, 61.1% and 49.6%, respectively. Resistance in the surface water to the same antibiotics was 87.1%, 24.1%, 7.6% and 12.9%, respectively. Overall, 86% of litter isolates, 58% of surface water isolates and 100% of ground water isolates were resistant to more than one antibiotic. Fifty-four different resistance patterns were recognised in isolates obtained from litter and environmental samples and several E. faecium and E. faecalis isolates from litter and environment samples shared the same resistance pattern. Multiple antibiotic resistant (MAR indices calculated to assess health risks due to the presence of resistant enterococci suggested an increased presence of antibiotics in surface water, likely from poultry sources as no other wastewater contributions in the area were documented.

  4. Protein-protein interactions between proteins of Citrus tristeza virus isolates.

    Science.gov (United States)

    Nchongboh, Chofong Gilbert; Wu, Guan-Wei; Hong, Ni; Wang, Guo-Ping

    2014-12-01

    Citrus tristeza virus (CTV) is one of the most devastating pathogens of citrus. Its genome is organized into 12 open reading frames (ORFs), of which ten ORFs located at the 3'-terminus of the genome have multiple biological functions. The ten genes at the 3'-terminus of the genome of a severe isolate (CTV-S4) and three ORFs (CP, CPm and p20) of three other isolates (N4, S45 and HB1) were cloned into pGBKT7 and pGADT7 yeast shuttle vectors. Yeast two-hybridization (Y2H) assays results revealed a strong self-interaction for CP and p20, and a unique interaction between the CPm of CTV-S4 (severe) and CP of CTV-N4 (mild) isolates. Bimolecular fluorescence complementation also confirmed these interactions. Analysis of the deletion mutants delineated the domains of CP and p20 self-interaction. Furthermore, the domains responsible for CP and p20 self-interactions were mapped at the CP amino acids sites 41-84 and p20 amino acids sites 1-21 by Y2H. This study provided new information on CTV protein interactions which will help for further understanding the biological functions.

  5. Promotion of bone growth by dietary soy protein isolate: Comparision with dietary casein, whey hydrolysate and rice protein isolate in growing female rats

    Science.gov (United States)

    The effects of different dietary protein sources(casein (CAS), soy protein isolate (SPI), whey protein hydrolysate (WPH) and rice protein isolate (RPI)) on bone were studied in intact growing female rats and in ovarectomized (OVX) rats showing sex steroid deficiency-induced bone loss. In addition, S...

  6. Increased Prevalence of Activated Protein C Resistance During ...

    African Journals Online (AJOL)

    levels of PCA ratio (measure of protein C resistance) of among our pregnant women since maternal mortality rate of the .... events in most developed countries where changes, which ... Height and weight are measured, and the body mass.

  7. Analysis of Genes Encoding Penicillin-Binding Proteins in Clinical Isolates of Acinetobacter baumannii ▿ †

    Science.gov (United States)

    Cayô, Rodrigo; Rodríguez, María-Cruz; Espinal, Paula; Fernández-Cuenca, Felipe; Ocampo-Sosa, Alain A.; Pascual, Álvaro; Ayala, Juan A.; Vila, Jordi; Martínez-Martínez, Luis

    2011-01-01

    There is limited information on the role of penicillin-binding proteins (PBPs) in the resistance of Acinetobacter baumannii to β-lactams. This study presents an analysis of the allelic variations of PBP genes in A. baumannii isolates. Twenty-six A. baumannii clinical isolates (susceptible or resistant to carbapenems) from three teaching hospitals in Spain were included. The antimicrobial susceptibility profile, clonal pattern, and genomic species identification were also evaluated. Based on the six complete genomes of A. baumannii, the PBP genes were identified, and primers were designed for each gene. The nucleotide sequences of the genes identified that encode PBPs and the corresponding amino acid sequences were compared with those of ATCC 17978. Seven PBP genes and one monofunctional transglycosylase (MGT) gene were identified in the six genomes, encoding (i) four high-molecular-mass proteins (two of class A, PBP1a [ponA] and PBP1b [mrcB], and two of class B, PBP2 [pbpA or mrdA] and PBP3 [ftsI]), (ii) three low-molecular-mass proteins (two of type 5, PBP5/6 [dacC] and PBP6b [dacD], and one of type 7 (PBP7/8 [pbpG]), and (iii) a monofunctional enzyme (MtgA [mtgA]). Hot spot mutation regions were observed, although most of the allelic changes found translated into silent mutations. The amino acid consensus sequences corresponding to the PBP genes in the genomes and the clinical isolates were highly conserved. The changes found in amino acid sequences were associated with concrete clonal patterns but were not directly related to susceptibility or resistance to β-lactams. An insertion sequence disrupting the gene encoding PBP6b was identified in an endemic carbapenem-resistant clone in one of the participant hospitals. PMID:21947403

  8. Characterization of lead-resistant river isolate Enterococcus faecalis and assessment of its multiple metal and antibiotic resistance.

    Science.gov (United States)

    Aktan, Yasin; Tan, Sema; Icgen, Bulent

    2013-06-01

    Contamination of surface waters has a direct impact on the public health of entire communities. Microorganisms inhabiting contaminated surface waters have developed mechanisms of coping with a variety of toxic metals and drugs. Investigations were carried out to isolate and identify lead-resistant bacteria from the river Kızılırmak along the city of Kırıkkale, Turkey. Of the 33 lead-resistant isolates, one isolate with a minimal inhibitory concentration of 1,200 mg L(-1) was isolated and identified as Enterococcus faecalis by using biochemical tests and 16S rRNA sequencing. Lead-resistant E. faecalis isolate was found out to be resistant to other heavy metals like aluminum, lithium, barium, chromium, iron, silver, tin, nickel, zinc, and strontium and to drugs like amikacin, aztreonam, and gentamicin. E. faecalis harbored four plasmids with the molecular sizes of 1.58, 3.06, 22.76, and 28.95 kb. Plasmid profile analyses of cured derivatives revealed that the lead resistance ability of E. faecalis was still existing despite the elimination of all the plasmids. Moreover, the antibiotic resistance pattern of the cured derivatives did not demonstrate any change from the parental strain. Our findings indicated that the lead resistance genes of E. faecalis were located on the chromosomal DNA rather than the plasmid.

  9. The role of active efflux in antibiotic - resistance of clinical isolates of Helicobacter pylori

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    Falsafi T

    2009-01-01

    Full Text Available Purpose: In gram-negative bacteria, active efflux pumps that excrete drugs can confer resistance to antibiotics however, in Helicobacter pylori this role is not well established. The purpose of this study is to evaluate the role of active efflux in resistance of H. pylori isolates to antibiotics. Materials and Methods: Twelve multiple antibiotic resistant (MAR isolates resistant to at least four antibiotics, including β-lactams, metronidazole, tetracycline, erythromycin, and ciprofloxacin; three resistant to only β-lactams, and two hyper-susceptible isolates, were obtained from screening of 96 clinical isolates of H. pylori . Their minimal inhibitory concentrations (MICs for antibiotics and ethidium-bromide (EtBr were compared in the presence- and absence of a proton-conductor, carbonyl cyanide-m chlorophenyl-hydrazone (CCCP using agar-dilution and disc diffusion. Drug accumulation studies for EtBr and antibiotics were assessed in the presence and absence of CCCP using spectrofluorometry. Results: MIC of EtBr for eight MAR-isolates was decreased two- to four-folds in the presence of CCCP, of which five showed reduced MICs for β-lactam, metronidazole, tetracycline, and ciprofloxacin with CCCP. Accumulation of EtBr by the MAR-isolates was rapid and not dependant on the pattern of multiple resistance. Antibiotic accumulation assay confirmed the presence of energy-dependant efflux of β-lactam, metronidazole, tetracycline, and ciprofloxacin, but no erythromycin in five MAR isolates. Energy-dependant efflux of EtBr or antibiotics was not observed for four MAR-isolates, and three isolates were resistant only to β-lactams. Conclusion: Energy-dependant efflux plays a role in the resistance of H. pylori clinical isolates to structurally unrelated antibiotics in a broadly specific multidrug efflux manner. Difference in the efflux potential of MAR isolates may be related to the presence or absence of functional efflux-pumps in diverse H. pylori

  10. Emergence of a colistin-resistant Escherichia coli clinical isolate harboring mcr-1 in Japan.

    Science.gov (United States)

    Tada, Tatsuya; Uechi, Kohei; Nakasone, Isamu; Shimada, Kayo; Nakamatsu, Masashi; Kirikae, Teruo; Fujita, Jiro

    2017-08-02

    The mcr-1 is a gene encoding a phosphoethanolamine transferase, which confers resistance to colistin by transferring phosphoethanolamine to lipid A. We describe here the emergence of a colistin-resistant Escherichia coli clinical isolate harboring plasmid-mediated mcr-1 in Japan. The isolate belonged to ST5702 and is suspected to come from livestock and transmitted to human. This is the first report of a clinical isolate harboring mcr-1 in Japan. Copyright © 2017. Published by Elsevier Ltd.

  11. "Emergence of Multidrug Resistant Strains of Escherichia coli Isolated from Urinary Tract Infections"

    OpenAIRE

    R Moniri; Khorshidi, A; H Akbari

    2003-01-01

    The emergence of multidrug resistant strains of Escherichia coli has complicated treatment decision and may lead to treatment failures. From April to November 2001 we prospectively evaluated the prevalence of resistance to trimethoprim-sulfamethoxazole (SXT), gentamicin, cephalothin, ciprofloxacin, and nitrofurantoin in 220 Escherichia coli isolates from patients with urinary tract infections in kashan, Iran. To assess the current breadth of multidrug resistance among urinary isolates of E. c...

  12. Biosurfactant and Heavy Metal Resistance Activity of Streptomyces spp. Isolated from Saltpan Soil

    OpenAIRE

    Lakshmipathy Deepika; Krishnan Kannabiran

    2010-01-01

    Actinomycetes were isolated from the marine soil samples collected at the Ennore saltpan and were screened for biosurfactant and heavy metal resistance activity. Biosurfactant activity was evaluated by haemolysis, drop collapsing test and lipase production. Similarly heavy metal resistance was determined by tube method and agar diffusion method. Among them, two actinomycetes isolates VITDDK1 and VITDDK2 exhibited significant biosurfactant and heavy metal resistance activity. Based on the Hide...

  13. Carbapenemase Genes among Multidrug Resistant Gram Negative Clinical Isolates from a Tertiary Hospital in Mwanza, Tanzania

    Directory of Open Access Journals (Sweden)

    Martha F. Mushi

    2014-01-01

    Full Text Available The burden of antimicrobial resistance (AMR is rapidly growing across antibiotic classes, with increased detection of isolates resistant to carbapenems. Data on the prevalence of carbapenem resistance in developing countries is limited; therefore, in this study, we determined the prevalence of carbapenemase genes among multidrug resistant gram negative bacteria (MDR-GNB isolated from clinical specimens in a tertiary hospital in Mwanza, Tanzania. A total of 227 MDR-GNB isolates were analyzed for carbapenem resistance genes. For each isolate, five different PCR assays were performed, allowing for the detection of the major carbapenemase genes, including those encoding the VIM-, IMP-, and NDM-type metallo-beta-lactamases, the class A KPC-type carbapenemases, and the class D OXA-48 enzyme. Of 227 isolates, 80 (35% were positive for one or more carbapenemase gene. IMP-types were the most predominant gene followed by VIM, in 49 (21.59% and 28 (12% isolates, respectively. Carbapenemase genes were most detected in K. pneumoniae 24 (11%, followed by P. aeruginosa 23 (10%, and E. coli with 19 isolates (8%. We have demonstrated for the first time a high prevalence of MDR-GNB clinical isolates having carbapenem resistance genes in Tanzania. We recommend routine testing for carbapenem resistance among the MDR-GNB particularly in systemic infections.

  14. Antibiotic resistance among cultured bacterial isolates from bioethanol fermentation facilities across the United States.

    Science.gov (United States)

    Murphree, Colin A; Heist, E Patrick; Moe, Luke A

    2014-09-01

    Bacterial contamination of fuel ethanol fermentations by lactic acid bacteria (LAB) can have crippling effects on bioethanol production. Producers have had success controlling bacterial growth through prophylactic addition of antibiotics to fermentors, yet concerns have arisen about antibiotic resistance among the LAB. Here, we report on mechanisms used by 32 LAB isolates from eight different US bioethanol facilities to persist under conditions of antibiotic stress. Minimum inhibitory concentration assays with penicillin, erythromycin, and virginiamycin revealed broad resistance to each of the antibiotics as well as high levels of resistance to individual antibiotics. Phenotypic assays revealed that antibiotic inactivation mechanisms contributed to the high levels of individual resistances among the isolates, especially to erythromycin and virginiamycin, yet none of the isolates appeared to use a β-lactamase. Biofilm formation was noted among the majority of the isolates and may contribute to persistence under low levels of antibiotics. Nearly all of the isolates carried at least one canonical antibiotic resistance gene and many carried more than one. The erythromycin ribosomal methyltransferase (erm) gene class was found in 19 of 32 isolates, yet a number of these isolates exhibit little to no resistance to erythromycin. The erm genes were present in 15 isolates that encoded more than one antibiotic resistance mechanism, suggestive of potential genetic linkages.

  15. Antimicrobial resistance in Salmonella enterica Serovar Typhi isolates from Bangladesh, Indonesia, Taiwan, and Vietnam.

    Science.gov (United States)

    Chiou, Chien-Shun; Lauderdale, Tsai-Ling; Phung, Dac Cam; Watanabe, Haruo; Kuo, Jung-Che; Wang, Pei-Jen; Liu, Yen-Yi; Liang, Shiu-Yun; Chen, Pei-Chen

    2014-11-01

    We characterized Salmonella enterica serovar Typhi isolates from Bangladesh, Indonesia, Taiwan, and Vietnam to investigate their genetic relatedness and antimicrobial resistance. The isolates from Bangladesh and Vietnam were genetically closely related but were distant from those from Indonesia and Taiwan. All but a few isolates from Indonesia and Taiwan were susceptible to all antimicrobials tested. The majority of isolates from Bangladesh and Vietnam were multidrug resistant (MDR) and belonged to the widespread haplotype H58 clone. IncHI1 plasmids were detected in all MDR S. Typhi isolates from Vietnam but in only 15% of MDR isolates from Bangladesh. Resistance genes in the majority of MDR S. Typhi isolates from Bangladesh should reside in the chromosome. Among the isolates from Bangladesh, 82% and 40% were resistant to various concentrations of nalidixic acid and ciprofloxacin, respectively. Several resistance mechanisms, including alterations in gyrase A, the presence of QnrS, and enhanced efflux pumps, were involved in the reduced susceptibility and resistance to fluoroquinolones. Intensive surveillance is necessary to monitor the spread of chromosome-mediated MDR and fluoroquinolone-resistant S. Typhi emerging in Bangladesh.

  16. Carbapenemase Genes among Multidrug Resistant Gram Negative Clinical Isolates from a Tertiary Hospital in Mwanza, Tanzania

    Science.gov (United States)

    Mushi, Martha F.; Mshana, Stephen E.; Imirzalioglu, Can; Bwanga, Freddie

    2014-01-01

    The burden of antimicrobial resistance (AMR) is rapidly growing across antibiotic classes, with increased detection of isolates resistant to carbapenems. Data on the prevalence of carbapenem resistance in developing countries is limited; therefore, in this study, we determined the prevalence of carbapenemase genes among multidrug resistant gram negative bacteria (MDR-GNB) isolated from clinical specimens in a tertiary hospital in Mwanza, Tanzania. A total of 227 MDR-GNB isolates were analyzed for carbapenem resistance genes. For each isolate, five different PCR assays were performed, allowing for the detection of the major carbapenemase genes, including those encoding the VIM-, IMP-, and NDM-type metallo-beta-lactamases, the class A KPC-type carbapenemases, and the class D OXA-48 enzyme. Of 227 isolates, 80 (35%) were positive for one or more carbapenemase gene. IMP-types were the most predominant gene followed by VIM, in 49 (21.59%) and 28 (12%) isolates, respectively. Carbapenemase genes were most detected in K. pneumoniae 24 (11%), followed by P. aeruginosa 23 (10%), and E. coli with 19 isolates (8%). We have demonstrated for the first time a high prevalence of MDR-GNB clinical isolates having carbapenem resistance genes in Tanzania. We recommend routine testing for carbapenem resistance among the MDR-GNB particularly in systemic infections. PMID:24707481

  17. Short communication: Genetic characterization of antimicrobial resistance in Acinetobacter isolates recovered from bulk tank milk.

    Science.gov (United States)

    Tamang, M D; Gurung, M; Nam, H M; Kim, S R; Jang, G C; Jung, S C; Lim, S K

    2014-02-01

    A total of 176 Acinetobacter isolates, including 57 Acinetobacter baumannii originally obtained from 2,287 bulk tank milk (BTM) samples in Korea was investigated for the genetic basis of antimicrobial resistance using molecular methods. In addition, the occurrence and cassette content of integrons were examined and the genetic diversity of A. baumannii strains identified was evaluated. Aminoglycoside-modifying enzyme genes were detected in 15 (88.2%) of the 17 aminoglycoside-resistant Acinetobacter isolates tested. The most common aminoglycoside-modifying enzyme gene identified was adenylyltransferase gene aadB (n = 9), followed by phosphotransferase genes aphA6 (n = 7) and aphA1 (n = 5). Of the 31 isolates resistant to tetracycline, tet(39) was detected in 20 of them. The genetic basis of resistance to sulfonamide was identified in 15 (53.6%) of 28 trimethoprim-sulfamethoxazole-resistant isolates and 9 (32.1%) of them carried both sul1 and sul2 genes. A blaADC-7-like gene was detected in 1 β-lactam-resistant A. baumannii. Furthermore, class 1 integron was identified in 11 Acinetobacter isolates. Two gene cassettes dfrA15, conferring resistance to trimethoprim, and aadA2, conferring resistance to aminoglycosides, were identified in 8 Acinetobacter isolates. None of the isolates was positive for class 2 or class 3 integrons. Pulsed-field gel electrophoresis revealed that most of the A. baumannii strains from BTM samples were genetically diverse, indicating that the occurrence of A. baumannii strains in BTM was not the result of dissemination of a single clone. Elucidation of resistance mechanisms associated with the resistance phenotype and a better understanding of resistance genes may help in the development of strategies to control infections, such as mastitis, and to prevent further dissemination of antibiotic resistance genes. To the best of our knowledge, this is the first report of molecular characterization of antimicrobial-resistant Acinetobacter spp. from

  18. Evaluation of Surrogate Disk Tests for Detection of Ciprofloxacin and Levofloxacin Resistance in Clinical Isolates of Salmonella enterica

    Science.gov (United States)

    Deak, Eszter; Skov, Robert; Hindler, Janet A.

    2015-01-01

    Detection of fluoroquinolone resistance in Salmonella enterica has become increasingly difficult due to evolving resistance mechanisms to this antimicrobial class in this organism. We evaluated two quinolone disks and five fluoroquinolone disks for their ability to act as a surrogate agent for the detection of fluoroquinolone resistance in a collection of 136 S. enterica isolates, including 111 with intermediate or resistant ciprofloxacin MICs mediated by a variety of resistance mechanisms. Ciprofloxacin, ofloxacin, and pefloxacin disks detected all isolates resistant to ciprofloxacin (0% very major error) and yielded false resistance (major error) in 8, 4, and 12% of susceptible isolates, respectively. Ciprofloxacin and pefloxacin provided clearer differentiation of susceptible and resistant isolates. PMID:26292293

  19. Class 1 integron-mediated antibiotic resistance in Salmonella enterica strains isolated in Tehran, Iran

    Directory of Open Access Journals (Sweden)

    Reza Ranjbar

    2014-03-01

    Conclusion: Our findings showed that integrons class 1 were widely distributed among Salmonella enterica isolates. Surveillance and monitoring of antimicrobial drug resistance, including screening for integrons as likely indicators of drug resistance and acquisition of new resistance traits, are necessary steps in planning effective strategies for containing this phenomenon within foodborne infection organisms.

  20. ERG11 mutations associated with azole resistance in Candida albicans isolates from vulvovaginal candidosis patients

    Institute of Scientific and Technical Information of China (English)

    Bin; Wang; Li-Hua; Huang; Ji-Xue; Zhao; Man; Wei; Hua; Fang; Dan-Yang; Wang; Hong-Fa; Wang; Ji-Gang; Yin; Mei; Xiang

    2015-01-01

    Objective: To investigate the azole susceptibility of Candida albicans(C.albicans)from vulvovaginal candidosis patients and to analyze the relationship between ERG11 gene mutations in these isolates and azole resistance.Methods: Three hundred and two clinical isolates of Candida species were collected.Azole susceptibility was tested in vitro in microdilution studies. The ERG11 genes of 17 isolates of C. albicans(2 susceptibles, 5 dose-dependent resistants and 10 resistants) were amplified and sequenced.Results: Of the 302 isolates collected, 70.2% were C. albicans, of which 8.5%, 3.8% and4.2% were resistant to fluconazole, itraconazole and voriconazole, respectively. In total,27 missense mutations were detected in ERG11 genes from resistant/susceptible dosedependent isolates. Among them, Y132 H, A114 S, and Y257 H substitutions were most prevalent and were known to cause fluconazole resistance. G464 S and F72 S also have been proved to cause fluconazole resistance. Two novel substitutions(T285A, S457P) in hotspot regions were identified.Conclusions: Twenty seven mutations in the ERG11 gene were identified in azoleresistant C. albicans isolates, which indicated a possible relation with the increase in resistance to azole drugs and the recurrence of vulvovaginal candidosis. The relationship of two novel substitutions(T285A, S457P) with fluconazole resistance needs to be further verified by site-directed mutagenesis.

  1. Physical properties, molecular structures and protein quality of texturized whey protein isolate: effect of extrusion temperature

    Science.gov (United States)

    Extrusion is a powerful food processing operation, which utilizes high temperature and high shear force to produce a product with unique physical and chemical characteristics. Texturization of whey protein isolate (WPI) through extrusion for the production of protein fortified snack foods has provid...

  2. Mechanisms, molecular and sero-epidemiology of antimicrobial resistance in bacterial respiratory pathogens isolated from Japanese children

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    Sunakawa Keisuke

    2007-08-01

    Full Text Available Abstract Background The clinical management of community-acquired respiratory tract infections (RTIs is complicated by the increasing worldwide prevalence of antibacterial resistance, in particular, β-lactam and macrolide resistance, among the most common causative bacterial pathogens. This study aimed to determine the mechanisms and molecular- and sero-epidemiology of antibacterial resistance among the key paediatric respiratory pathogens in Japan. Methods Isolates were collected at 18 centres in Japan during 2002 and 2003 from children with RTIs as part of the PROTEKT surveillance programme. A proportion of Haemophilus influenzae isolates was subjected to sequencing analysis of the ftsI gene; phylogenetic relatedness was assessed using multilocus sequence typing. Streptococcus pneumoniae isolates were screened for macrolide-resistance genotype by polymerase chain reaction and serotyped using the capsular swelling method. Susceptibility of isolates to selected antibacterials was performed using CLSI methodology. Results and Discussion Of the 557 H. influenzae isolates collected, 30 (5.4% were β-lactamase-positive [BL+], 115 (20.6% were BL-nonproducing ampicillin-resistant (BLNAR; MIC ≥ 4 mg/L and 79 (14.2% were BL-nonproducing ampicillin-intermediate (BLNAI; MIC 2 mg/L. Dabernat Group III penicillin binding protein 3 (PBP3 amino acid substitutions in the ftsI gene were closely correlated with BLNAR status but phylogenetic analysis indicated marked clonal diversity. PBP mutations were also found among BL+ and BL-nonproducing ampicillin-sensitive isolates. Of the antibacterials tested, azithromycin and telithromycin were the most active against H. influenzae (100% and 99.3% susceptibility, respectively. A large proportion (75.2% of the 468 S. pneumoniae isolates exhibited macrolide resistance (erythromycin MIC ≥ 1 mg/L; erm(B was the most common macrolide resistance genotype (58.8%, followed by mef(A (37.2%. The most common pneumococcal

  3. Molecular characterization of boscalid resistance in field isolates of Botrytis cinerea from apple.

    Science.gov (United States)

    Yin, Y N; Kim, Y K; Xiao, C L

    2011-08-01

    Botrytis cinerea isolates obtained from apple orchards were screened for resistance to boscalid. Boscalid-resistant (BosR) isolates were classified into four phenotypes based on the levels of the concentration that inhibited fungal growth by 50% relative to control. Of the 220 isolates tested, 42 were resistant to boscalid, with resistant phenotypes ranging from low to very high resistance. There was cross resistance between boscalid and carboxin. Analysis of partial sequences of the iron-sulfur subunit of succinate dehydrogenase gene in B. cinerea (BcSdhB) from 13 BosR and 9 boscalid-sensitive (BosS) isolates showed that point mutations in BcSdhB leading to amino acid substitutions at the codon position 272 from histidine to either tyrosine (H272Y) or arginine (H272R) were correlated with boscalid resistance. Allele-specific polymerase chain reaction (PCR) analysis of 66 BosR isolates (including 24 additional isolates obtained from decayed apple fruit) showed that 19 carried the point mutation H272Y and 46 had the point mutation H272R, but 1 BosR isolate gave no amplification product. Analysis of the BcSdhB sequence of this isolate revealed a different point mutation at codon 225, resulting in a substitution of proline (P) by phenylalanine (F) (P225F). The results indicated that H272R/Y in BcSdhB were the dominant genotypes of mutants in field BosR isolates from apple. A multiplex allele-specific PCR assay was developed to detect point mutations H272R/Y in a single PCR amplification. Levels of boscalid resistance ranged from low to very high within isolates carrying either the H272R or H272Y mutation, indicating that, among BosR isolates, different BosR phenotypes (levels of resistance) were not associated with particular types of point mutations (H272R versus H272Y) in BcSdhB. Analysis of genetic relationships between 39 BosR and 56 BosS isolates based on three microsatellite markers showed that 39 BosR isolates and 30 BosS isolates were clustered into two groups

  4. Occurrence of virulence factors and antimicrobial resistance in Pasteurella multocida strains isolated from slaughter cattle in Iran.

    Science.gov (United States)

    Khamesipour, Faham; Momtaz, Hassan; Azhdary Mamoreh, Morteza

    2014-01-01

    A total of 30 Pasteurella multocida strains isolated from 333 pneumonic and apparently health slaughter cattle were examined for capsule biosynthesis genes and 23 virulence-associated genes by polymerase chain reaction (PCR). The disc diffusion technique was used to determine antimicrobial resistance profiles among the isolates. Of the isolates, 23 belonged to capsular type A, 5 to capsular type D and two isolates were untypeable. The distribution of the capsular types in pneumonic lungs and in apparently health lungs was statistically similar. All virulence genes tested were detected among the isolates derived from pneumonic lungs; whereas isolates derived from apparently health lungs carried 16 of the 23 genes. The frequently detected genes among isolates from pneumonic lungs were exbD, hgbA, hgbB, ompA, ompH, oma87, and sodC; whereas tadD, toxA, and pmHAS genes occurred less frequently. Most of the adhesins and superoxide dismutases; and all of the iron acquisition and protectin proteins occurred at significantly (p ≤ 0.05) higher frequencies in isolates from pneumonic lungs. Isolates from apparently healthy lungs didn't carry the following genes; hsf-1, hsf-2, tadD, toxA, nanB, nanH, and pmHAS. One adhesion (hsf-1) and two iron acquisition (exbD and tonB) genes occurred at significantly (p ≤ 0.05) higher frequencies among capA isolates. All the P. multocida isolates were susceptible to ciprofloxacin, co-trimoxazole, doxycycline, enrofloxacin, nitrofurantoin, and tetracyclines. Different proportions of the isolates were however resistant to ampicillin, amoxicillin, erythromycin, lincomycin, penicillin, rifampin, streptomycin, and florfenicol. Our results reveal presence of virulence factors (VFs) in P. multocida strains isolated from symptomatic and asymptomatic bovids. A higher frequency of the factors among isolates from symptomatic study animals may suggest their role in pathogenesis of P. multocida-associated bovine respiratory disease (BRD). The results

  5. Antimicrobial resistance genes in Actinobacillus pleuropneumoniae, Haemophilus parasuis and Pasteurella multocida isolated from Australian pigs.

    Science.gov (United States)

    Dayao, Dae; Gibson, J S; Blackall, P J; Turni, C

    2016-07-01

    To identify genes associated with the observed antimicrobial resistance in Actinobacillus pleuropneumoniae, Haemophilus parasuis and Pasteurella multocida isolated from Australian pigs. Isolates with known phenotypic resistance to β-lactams, macrolides and tetracycline were screened for the presence of antimicrobial resistance genes. A total of 68 A. pleuropneumoniae, 62 H. parasuis and 20 P. multocida isolates exhibiting phenotypic antimicrobial resistance (A. pleuropneumoniae and P. multocida) or elevated minimal inhibitory concentrations (MICs) (H. parasuis) to any of the following antimicrobial agents - ampicillin, erythromycin, penicillin, tetracycline, tilmicosin and tulathromycin - were screened for a total of 19 associated antimicrobial resistance genes (ARGs) by PCR. The gene bla ROB-1 was found in all ampicillin- and penicillin-resistant isolates, but none harboured the bla TEM-1 gene. The tetB gene was found in 76% (74/97) of tetracycline-resistant isolates, 49/53 A. pleuropneumoniae, 17/30 H. parasuis and 8/14 P. multocida. One A. pleuropneumoniae isolate harboured the tetH gene, but none of the 97 isolates had tetA, tetC, tetD, tetE, tetL, tetM or tetO. A total of 92 isolates were screened for the presence of macrolide resistance genes. None was found to have ermA, ermB, ermC, erm42, mphE, mefA, msrA or msrE. The current study has provided a genetic explanation for the resistance or elevated MIC of the majority of isolates of Australian porcine respiratory pathogens to ampicillin, penicillin and tetracycline. However, the macrolide resistance observed by phenotypic testing remains genetically unexplained and further studies are required. © 2016 Australian Veterinary Association.

  6. ABC-F Proteins Mediate Antibiotic Resistance through Ribosomal Protection.

    Science.gov (United States)

    Sharkey, Liam K R; Edwards, Thomas A; O'Neill, Alex J

    2016-03-22

    Members of the ABC-F subfamily of ATP-binding cassette proteins mediate resistance to a broad array of clinically important antibiotic classes that target the ribosome of Gram-positive pathogens. The mechanism by which these proteins act has been a subject of long-standing controversy, with two competing hypotheses each having gained considerable support: antibiotic efflux versus ribosomal protection. Here, we report on studies employing a combination of bacteriological and biochemical techniques to unravel the mechanism of resistance of these proteins, and provide several lines of evidence that together offer clear support to the ribosomal protection hypothesis. Of particular note, we show that addition of purified ABC-F proteins to anin vitrotranslation assay prompts dose-dependent rescue of translation, and demonstrate that such proteins are capable of displacing antibiotic from the ribosomein vitro To our knowledge, these experiments constitute the first direct evidence that ABC-F proteins mediate antibiotic resistance through ribosomal protection.IMPORTANCEAntimicrobial resistance ranks among the greatest threats currently facing human health. Elucidation of the mechanisms by which microorganisms resist the effect of antibiotics is central to understanding the biology of this phenomenon and has the potential to inform the development of new drugs capable of blocking or circumventing resistance. Members of the ABC-F family, which includelsa(A),msr(A),optr(A), andvga(A), collectively yield resistance to a broader range of clinically significant antibiotic classes than any other family of resistance determinants, although their mechanism of action has been controversial since their discovery 25 years ago. Here we present the first direct evidence that proteins of the ABC-F family act to protect the bacterial ribosome from antibiotic-mediated inhibition. Copyright © 2016 Sharkey et al.

  7. Molecular characterization of fluoroquinolone resistance in nontypeable Haemophilus influenzae clinical isolates.

    Science.gov (United States)

    Puig, Carmen; Tirado-Vélez, José Manuel; Calatayud, Laura; Tubau, Fe; Garmendia, Junkal; Ardanuy, Carmen; Marti, Sara; de la Campa, Adela G; Liñares, Josefina

    2015-01-01

    Nontypeable Haemophilus influenzae (NTHi) is a common cause of respiratory infections in adults, who are frequently treated with fluoroquinolones. The aims of this study were to characterize the genotypes of fluoroquinolone-resistant NTHi isolates and their mechanisms of resistance. Among 7,267 H. influenzae isolates collected from adult patients from 2000 to 2013, 28 (0.39%) were ciprofloxacin resistant according to Clinical and Laboratory Standards Institute (CLSI) criteria. In addition, a nalidixic acid screening during 2010 to 2013 detected five (0.23%) isolates that were ciprofloxacin susceptible but nalidixic acid resistant. Sequencing of their quinolone resistance-determining regions and genotyping by pulse-field gel electrophoresis and multilocus sequence typing of the 25 ciprofloxacin-resistant isolates available and all 5 nalidixic acid-resistant isolates were performed. In the NTHi isolates studied, two mutations producing changes in two GyrA residues (Ser84, Asp88) and/or two ParC residues (Ser84, Glu88) were associated with increased fluoroquinolone MICs. Strains with one or two mutations (n = 15) had ciprofloxacin and levofloxacin MICs of 0.12 to 2 μg/ml, while those with three or more mutations (n = 15) had MICs of 4 to 16 μg/ml. Long persistence of fluoroquinolone-resistant strains was observed in three chronic obstructive pulmonary disease patients. High genetic diversity was observed among fluoroquinolone-resistant NTHi isolates. Although fluoroquinolones are commonly used to treat respiratory infections, the proportion of resistant NTHi isolates remains low. The nalidixic acid disk test is useful for detecting the first changes in GyrA or in GyrA plus ParC among fluoroquinolone-susceptible strains that are at a potential risk for the development of resistance under selective pressure by fluoroquinolone treatment. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  8. Resistance to antimicrobial agents of Campylobacter spp. strains isolated from animals in Poland.

    Science.gov (United States)

    Krutkiewicz, A; Sałamaszyńska-Guz, A; Rzewuska, M; Klimuszko, D; Binek, M

    2009-01-01

    A total of 69 Campylobacter jejuni and 16 Campylobacter coli strains isolated from chicken, dog and pig stool samples were characterized based on their resistance to five antimicrobial agents and on plasmid pTet profiles. Antimicrobials used in this study were: amoxicillin/clavulanic acid, ciprofloxacin, erythromycin, tetracycline and trimethoprim/sulfamethoxazole. Among the isolates studied, 91.7% were resistant to one or more antimicrobial agent. The highest level of resistance for the whole test group was to trimethoprim/sulfamethoxazole (57.6%), followed by ciprofloxacin (44.2%) and tetracycline (20%). All isolates were susceptible to amoxicillin/clavulanic acid. Strains isolated from chickens were susceptible to erythromycin. Few erythromycin-resistant strains were isolated from dogs and pigs (5.8%). C. coli strains exhibited a higher antibiotic resistance than C. jejuni strains, excluding resistance to trimethoprim/sulfamethoxazole. The pTet plasmid harboring the tet(O) gene was detected in 14 Campylobacter spp. strains. Our studies demonstrate that the majority (71.4%) of tetracycline-resistant isolates carry a plasmid-borne tet(O) gene, particularly strains for which the minimum inhibitory concentration (MIC) are > or = 256 microg/ml. In conclusion, we have found high-level trimethoprim/sulfamethoxazole, ciprofloxacin and tetracycline resistance in Polish strains isolated from different sources. This study has demonstrated that resistance of Campylobacter species differs depending on both the bacterial species and animal origins. All strains that displayed resistance to four antimicrobial agents were isolated from pigs. Localization of the tet(O) gene on either plasmid or chromosome was not found to be correlated with tetracycline resistance.

  9. Prevalence and Fluoroquinolone Resistance Pattern in Escherichia coli Isolates of Urinary Tract Infection (UTI Patients

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    Tippireddypalli Gururaju,

    2015-04-01

    Full Text Available Background: Urinary tract infections (UTIs are among the most common infectious diseases all over the world. Recent studies reported an increased antibiotic resistance in Escherichia coli, primary causative agent of UTI. The resistance has emerged even to more potent antimicrobial agents like fluoroquinolones. Objectives: The present study was undertaken to evaluate the prevalence and resistance pattern of E.coli causing UTIs in patients admitted to a tertiary care hospital in South India, with reference to fluoroquinolones. Material and Methods: A total of 278 selected urine samples of urinary tract infections were processed for E.coli culture using standard methods. For these urinary E. coli isolates, susceptibility to various antibiotics including fluoroquinolones was checked by Kirby Bauer disk diffusion method according CLSI criteria. Final resistance to fluoroquinolones isolates was analyzed. Results: Out of the 278 selected UTI clinical isolates 148 (54% showed ciprofloxacin sensitive and 130 (46% clinical isolates are ciprofloxacin resistant. Of the 130 ciprofloxacin resistant urinary isolates of E. coli subjected to susceptibility test for increased generation of fluoroquinolone drugs, the pattern of resistance noticed as levofloxacin (2nd generation 79%, gatifloxacin (3rd generation 77% and moxifloxacin (4th generation 75%, respectively. The fluoroquinolone resistance in UTI clinical isolates was decreasing with increasing generations of fluoroquinolone. Quinolone drug resistance in clinical isolates was increasing with age and hospitalized patients. Conclusion: Study showed an increased fluoroquinolone resistance among uropathogenic E. coli isolates of UTI. These increased antibiotic resistance trends in UTI patients indicated that it is imperative to rationalize the use of antimicrobials and to use them conservatively.

  10. Enzymes associated with protein bodies isolated from ungerminated barley seeds.

    Science.gov (United States)

    Ory, R L; Henningsen, K W

    1969-11-01

    Protein bodies were isolated intact from dormant barley seeds, Hordeum vulgare, var. Kenia, by a combination of buffer extractions and centrifugations over a sucrose gradient. Examination of the protein bodies pellet in the electron microscope shows 2 types of protein bodies in a wide variation of sizes. The majority of them stain evenly with osmium, are contained within a single membrane, and have no other structural components. The other type, mostly the larger particles, has a fine structure of orderly dark and light-stained layers attached to the protein bodies. Two acid hydrolases are associated with these particles: acid phosphatase activity, specific for sodium phytate but inactive on beta-glycerol phosphate, glucose 1-phosphate, fructose 1,6-diphosphate and adenosine triphosphate; and acid protease activity.

  11. Analysis of resistance, cross-resistance and antimicrobial combinations for Pseudomonas aeruginosa isolates from 1997 to 2009.

    Science.gov (United States)

    Master, Ronald N; Clark, Richard B; Karlowsky, James A; Ramirez, Julio; Bordon, Jose M

    2011-10-01

    Pseudomonas aeruginosa is a nosocomial and community-acquired pathogen associated with considerable patient morbidity and mortality. Multidrug resistance in P. aeruginosa is a concern owing to the limited therapeutic options available to treat infections due to this organism. In this study, rates of antimicrobial resistance of P. aeruginosa isolates collected by The Surveillance Network Database-USA (Eurofins Medinet, Chantilly, VA) from 1997 to 2009 were examined. The patient population and specimens were stratified according to patient setting and age as well as specimen source. Multidrug resistance was defined as resistance to three or more of the following antimicrobial agents: aztreonam; cefepime; ciprofloxacin; imipenem; gentamicin; and piperacillin/tazobactam (TZP). A total of 924740 P. aeruginosa isolates were examined in this study. Changes in resistance rates to individual antimicrobial agents were resistant P. aeruginosa to four, five and six antimicrobial agents. For isolates resistant to imipenem, aztreonam and gentamicin, ciprofloxacin had the highest cross-resistance rates. The greatest coverage against P. aeruginosa was by the combination of TZP plus amikacin (94%) followed by aztreonam plus amikacin (90%). Pseudomonas aeruginosa resistance rates remained steady or minimally declined to all antimicrobials from 1997 to 2009. Amongst the β-lactams, TZP has the greatest activity against P. aeruginosa. Copyright © 2011 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  12. Analysis on drug-resistance and molecular epidemiology of Acinetobacter baumannii isolated from the clinical samples in two Chinese hospitals

    Institute of Scientific and Technical Information of China (English)

    WEI FENG SHI; ZHI MI HUANG; NING XU

    2006-01-01

    In the present study, the drug-resistance genes encoding β-lactamases, aminoglycoside modifying enzymes, DNA topoisomerases and integron as well as their molecular epidemiology were investigated by means of analyzing the drug-resistance and molecular epidemiology of Acinebacter baumannii isolated from the clinical samples in two hospitals in Changzhou and Huzhou city of Jiangsu and Zhejiang province from July 2000 to March 2005. The minimal inhibitory concentrations (MICs) of these 307 isolates were detected by automatic microbiological system, and 35 strains against 5-fluoroquinolones were performed by agar dilution assay. Meanwhile, the resistant genes in 80 isolates were amplified by PCR with identification by DNA sequencer. It was found that most of the 307 isolates of A. baumannii were resistant to multiple antibiotics tested, in which the resistance rates of the isolates against piperacillin, piperacillin/tazobactam, amoxacillin/clavulanic acid, cefotaxime, ceftazidime,cefepime, gentamicin, amikacin, ciprofloxacin, chloramphenicol and sulfamethoxazole/trimethoprim were all above 35%, but those of imipenem and meropenem were quite low, ranged only 2.6% and 3.3 %. In addition, it was also demonstrated that the positive rates of TEM and SHV β-lactamase genes accounted for 93.8% and 22.5% respectively, and those of the aminoglycoside-modifying enzyme genes including aacC1, aacC2, aacC3, aacC4, aacC4A, aphA6, ant(2")-Ⅰ and ant(3")-Ⅰ were 58.8%, 8.8%, 7.5%, 28.8%, 45.0%, 2.5%, 28.8% and 65.0% respectively. The mutations in the quinolone-resistant determining region (QRDR) of gyrA and parC genes indicated that substitution in Ser-83 residue of GyrA protein was most frequently occurred among strains with MIC for ciprofloxacin of more than 4 μg/ml, whereas a double mutation at Ser-83 residue of gyrA and Ser-80 of parC was found in strains with MIC of ciprofloxacin of more than 8 μg/ml. As to the positive rates of class 1 integron (Int Ⅰ -1) and qacE△1-sul

  13. Diversity of Molecular Mechanisms Conferring Carbapenem Resistance to Pseudomonas aeruginosa Isolates from Saudi Arabia

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    Mohamed H. Al-Agamy

    2016-01-01

    Full Text Available Background. This study described various molecular and epidemiological characters determining antibiotic resistance patterns in Pseudomonas aeruginosa isolates. Methods. A total of 34 carbapenem-resistant P. aeruginosa clinical isolates were isolated from samples collected at a tertiary hospital in Riyadh, Saudi Arabia, from January to December 2011. Susceptibility testing, serotyping, molecular characterization of carbapenem resistance, and pulsed-field gel electrophoresis (PFGE were performed. Results. All isolates were resistant to ceftazidime, and more than half were highly resistant (minimum inhibitory concentration (MIC > 256 mg/L. Fifteen isolates had MIC values ≥64 mg/L for any of the carbapenems examined. Vietnamese extended-spectrum β-lactamase (VEB-1 (n=16/34 and oxacillinase (OXA-10 (n=14/34 were the most prevalent extended-spectrum β-lactamase and penicillinase, respectively. Verona imipenemase (VIM-1, VIM-2, VIM-4, VIM-11, and VIM-28 and imipenemase (IMP-7 variants were found in metallo-β-lactamase producers. A decrease in outer membrane porin gene (oprD expression was seen in nine isolates, and an increase in efflux pump gene (MexAB expression was detected in five isolates. Six serotypes (O:1, O:4, O:7, O:10, O:11, and O:15 were found among the 34 isolates. The predominant serotype was O:11 (16 isolates, followed by O:15 (nine isolates. PFGE analysis of the 34 carbapenem-resistant P. aeruginosa isolates revealed 14 different pulsotypes. Conclusions. These results revealed diverse mechanisms conferring carbapenem resistance to P. aeruginosa isolates from Saudi Arabia.

  14. ANTIBIOTICS RESISTANCE AND PUTATIVE VIRULENCE FACTORS OF AEROMONAS HYDROPHILA ISOLATED FROM ESTUARY

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    Olumide Adedokun Odeyemi

    2012-06-01

    Full Text Available This study aim to investigate antibiotics resistance profile and putative virulence factors of Aeromonas hydrophila isolated from estuary. Bacteria used for this study were isolated from water and sediment samples obtained from Sungai Melayu, Johor, Malaysia. Serially diluted 100 µL water and 1g sediment were inoculated on modified Rimler - Shott (mRS agar. Colonies with distinct cultural characteristics were picked for further studies. Isolates were tested for biofilm productions, protease enzyme and antibiotics resistance profile using agar well diffusion method against 10 commercial antibiotics. Congo Red Agar (CRA, Microplate and Standard Tube (ST methods were used for assessment of biofilm formation among the isolates while Skim Milk Agar was used for protease production. Sw.KMJ 3 and Sw.KMJ 9 produced black crystalline colonies on CRA. Six of the isolates were biofilm producers in ST method. Result of Microplate method, helped in grouping the isolates into weak (n = 8, moderate (n = 3 and strong producers (n = 4 at 540 nm wavelength. All the isolates were classified as weak ODc  ODi 0.1, moderate ODi = 0.1  0.12 and strong producers ODi  0.12 respectively at 540 nm wavelength. Antibiotics susceptibility test also revealed that all the isolates were resistant to between 6 and 10 antibiotics. Two isolates each were resistant to 6 (60 %, 7 (70 % and 9 (90 % antibiotics respectively. Eight of the isolates showed resistance to 8 (80 % antibiotics while only isolate Sw.KMJ-7 showed resistance to all the tested antibiotics. Sw.KMJ-3, Sw.KMJ-8 and Sw.KMJ-9 produced protease enzyme on SMA. The isolates were also found to be resistant to both antibiotics and heavy metals.

  15. Efficient isolation and elution of cellular proteins using aptamer-mediated protein precipitation assay.

    Science.gov (United States)

    Kim, Kiseok; Lee, SeungJin; Ryu, Sungho; Han, Dongil

    2014-05-23

    Protein precipitation is one of the most widely used methods for antigen detection and purification in biological research. We developed a reproducible aptamer-mediated magnetic protein precipitation method that is able to efficiently capture, purify and isolate the target proteins. We discovered DNA aptamers having individually high affinity and specificity against human epidermal growth factor receptor (EGFR) and human insulin receptor (INSR). Using aptamers and magnetic beads, we showed it is highly efficient technique to enrich endogenous proteins complex and is applicable to identify physiologically relevant protein-protein interactions with minimized nonspecific binding of proteins. The results presented here indicate that aptamers would be applicable as a useful and cost-effective tool to identify the presence of the particular target protein with their specific protein partners.

  16. Distribution of Flagella Secreted Protein and Integral Membrane Protein Among Campylobacter jejuni Isolated from Thailand

    Science.gov (United States)

    2011-01-01

    secreted protein and integral membrane protein among Campylobacter jejuni isolated from Thailand Piyarat Pootong 1·, Oralak Serichantalergs...Ladaporn Bodhidatta \\ Frederic Poly2, Patricia Guerry2 and Carl J Mason 1 Abstract Background: Campylobacter jejuni, a gram-negative bacterium, is a...groups of integral membrane protein. The significance of these different FspA variants to virulence requires further study. Background Campylobacter

  17. In vitro activity of telithromycin against Spanish Streptococcus pneumoniae isolates with characterized macrolide resistance mechanisms.

    Science.gov (United States)

    Morosini, M I; Cantón, R; Loza, E; Negri, M C; Galán, J C; Almaraz, F; Baquero, F

    2001-09-01

    The susceptibilities to telithromycin of 203 Streptococcus pneumoniae isolates prospectively collected during 1999 and 2000 from 14 different geographical areas in Spain were tested and compared with those to erythromycin A, clindamycin, quinupristin-dalfopristin, penicillin G, cefotaxime, and levofloxacin. Telithromycin was active against 98.9% of isolates (MICs, MICs at which 90% of isolates are inhibited being 0.06 microg/ml, irrespective of the resistance genotype. The corresponding values for erythromycin were 61.0% (MICs, 64 microg/ml. The erm(B) gene (macrolide-lincosamide-streptogramin B resistance phenotype) was detected in 36.4% (n = 74) of the isolates, which corresponded to 93.6% of erythromycin-intermediate and -resistant isolates, whereas the mef(A) gene (M phenotype [resistance to erythromycin and susceptibility to clindamycin and spiramycin without blunting]) was present in only 2.4% (n = 5) of the isolates. One of the latter isolates also carried erm(B). Interestingly, in one isolate for which the erythromycin MIC was 2 microg/ml, none of these resistance genes could be detected. Erythromycin MICs for S. pneumoniae erm(B)-positive isolates were higher (range, 0.5 to >64 microg/ml) than those for erm(B)- and mef(A)-negative isolates (range, 0.008 to 2 microg/ml). The corresponding values for telithromycin were lower for both groups, with ranges of 0.004 to 1 and 0.002 to 0.06 microg/ml, respectively. The erythromycin MIC was high for a large number of erm(B)-positive isolates, but the telithromycin MIC was low for these isolates. These results indicate the potential usefulness of telithromycin for the treatment of infections caused by erythromycin-susceptible and -resistant S. pneumoniae isolates when macrolides are indicated.

  18. Frequency of Resistance and Susceptible Bacteria Isolated from Houseflies

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    B Davari

    2010-12-01

    Conclusion: Houseflies collected from hospitals and slaughterhouse may be involved in the spread of drug resistant bacteria and may increase the potential of human exposure to drug resistant bacteria.

  19. Short report: polymorphisms in the chloroquine resistance transporter gene in Plasmodium falciparum isolates from Lombok, Indonesia.

    Science.gov (United States)

    Huaman, Maria Cecilia; Yoshinaga, Kazumi; Suryanatha, Aan; Suarsana, Nyoman; Kanbara, Hiroji

    2004-07-01

    The polymorphisms in the Plasmodium falciparum multidrug resistance 1 (pfmdr1) and P. falciparum chloroquine resistance transporter (pfcrt) genes, which are associated with chloroquine resistance, were examined in 48 P. falciparum isolates from uncomplicated malaria patients from the West Lombok District in Indonesia. The point mutation N86Y in pfmdr1 was present in 35.4% of the isolates and mutation K76T in pfcrt was found in all but one of the samples studied. Identified pfcrt haplotypes were mainly identical to the Papua New Guinea type S(agt)VMNT (42 of 48, 87.5%), and a few isolates had the Southeast Asia type CVIET (5 of 48, 10.4%). Moreover, one P. falciparum isolate harbored the K76N mutation, giving rise to the haplotype CVMNN, which was not previously reported in field isolates. Our findings suggest that chloroquine resistance in this area might have the same origin as in Papua New Guinea.

  20. Antimicrobial resistance of Staphylococcus pseudintermedius isolates from healthy dogs and dogs affected with pyoderma in Japan.

    Science.gov (United States)

    Onuma, Kenta; Tanabe, Taishi; Sato, Hisaaki

    2012-02-01

    Staphylococcus pseudintermedius strains were isolated from healthy dogs and dogs with pyoderma in 2000-2002 and 2009. All the isolates from dogs with pyoderma in 1999-2000 and from healthy dogs in 2000-2002 and 2009 were susceptible to cefalexin and/or other cephalosporins and oxacillin. However, 7.1-12.5 and 11.4% of S. pseudintermedius isolates from dogs with pyoderma in 2009 were resistant to cephalosporins and oxacillin, respectively. All S. pseudintermedius isolates from dogs with pyoderma in 1999-2000 and those from healthy dogs in 2000-2002 were susceptible to fluoroquinolones; however, 50% of the S. pseudintermedius strains isolated from dogs with pyoderma in 2009 and 30% of the S. pseudintermedius strains isolated from healthy dogs in 2009 were resistant to fluoroquinolones. Of the 21 oxacillin-resistant S. pseudintermedius (MRSP) isolates, 11 carried SCCmec type V and 10 carried hybrid SCCmec types II-III. Staphylococcus pseudintermedius strains that were resistant to only one of three fluoroquinolones had a mutation in the quinolone resistance determination region of grlA, whereas S. pseudintermedius strains that were resistant to two or more fluoroquinolones had mutations in the quinolone resistance determination regions of both grlA and gyrA.

  1. Antimicrobial resistance among Campylobacter jejuni isolated from raw poultry meat at retail level in Denmark

    DEFF Research Database (Denmark)

    Andersen, S. R.; Saadbye, P.; Shukri, Naseer Mahmoud

    2006-01-01

    Campylobacter jejuni isolated from raw poultry meat collected at retail shops in Denmark in the period 1996-2003 were tested for susceptibility to seven antimicrobial agents. The food samples consisted of raw chicken meat and other raw poultry meat of domestic or imported origin. The highest levels...... of resistance among C. jejuni were observed for tetracycline, nalidixic acid and ciprofloxacin, whereas macrolide resistance was rarely detected. C. jejuni originating from other poultry meat (mainly duck and turkey meat) exhibited the highest occurrences of antimicrobial resistance monitored; approximately one...... third of the isolates were tetracycline resistant (N=100). Among chicken meat isolates, the occurrence of tetracycline resistance was significantly higher (P jejuni isolated from imported chicken meat (N=88) than in C. jejuni from Danish chicken meat (N=367). The same tendency was observed...

  2. Whey protein isolate attenuates strength decline after eccentrically-induced muscle damage in healthy individuals

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    Cribb Paul J

    2010-09-01

    Full Text Available Abstract Background We examined the effects of short-term consumption of whey protein isolate on muscle proteins and force recovery after eccentrically-induced muscle damage in healthy individuals. Methods Seventeen untrained male participants (23 ± 5 yr, 180 ± 6 cm, 80 ± 11 kg were randomly separated into two supplement groups: i whey protein isolate (WPH; n = 9; or ii carbohydrate (CHO; n = 8. Participants consumed 1.5 g/kg.bw/day supplement (~30 g consumed immediately, and then once with breakfast, lunch, in the afternoon and after the evening meal for a period of 14 days following a unilateral eccentric contraction-based resistance exercise session, consisting of 4 sets of 10 repetitions at 120% of maximum voluntary contraction on the leg press, leg extension and leg flexion exercise machine. Plasma creatine kinase and lactate dehydrogenase (LDH levels were assessed as blood markers of muscle damage. Muscle strength was examined by voluntary isokinetic knee extension using a Cybex dynamometer. Data were analyzed using repeated measures ANOVA with an alpha of 0.05. Results Isometric knee extension strength was significantly higher following WPH supplementation 3 (P Conclusions The major finding of this investigation was that whey protein isolate supplementation attenuated the impairment in isometric and isokinetic muscle forces during recovery from exercise-induced muscle injury.

  3. Resistant plasmid profile analysis of multidrug resistant Escherichia coli isolated from urinary tract infections in Abeokuta, Nigeria.

    Science.gov (United States)

    Akingbade, O; Balogun, S; Ojo, D; Akinduti, P; Okerentugba, P O; Nwanze, J C; Okonko, I O

    2014-12-01

    Multi-drug resistant Escherichia coli has become a major threat and cause of many urinary tract infections (UTIs) in Abeokuta, Nigeria. This study was carried out to determine the resistant plasmids of multidrug resistant Escherichia coli isolated from (Urinary tract infections)UTIs in Abeokuta. A total of 120 Escherichia coli isolates were obtained from urine samples collected from patients attending inpatient and outpatient clinics presenting UTI; with their biodata. Antibiotics susceptibility was performed and multi-drug resistant isolates were selected for plasmid profiling. Plasmids were extracted by the alkaline lysis method, electrophoresed on 0.8% agarose gel and profiled using a gel-photo documentation system gel. Escherichia coli isolates obtained shows high resistance to cloxacillin (92.5%), amoxicillin (90.8%), ampicillin (90.8%), erythromycin (75.8%), cotrimoxazole (70.0%), streptomycin (70.0%) and tetracycline (68.3%) while 85.8% and 84.2% were susceptible to gentamycin and ceftazidime respectively. Sixteen Escherichia coli strains were observed to be resistant to more than two classes of antibiotics. The resistant plasmid DNA was detectable in 6(37.5%) of the 16 multidrug resistant Escherichia coli having single sized plasmids of the same weight 854bp and were all resistant to erythromycin, cefuroxime, cloxacillin, amoxicillin, ampicillin and cotrimoxazole. This study has highlighted the emergence of multidrug resistant R-plasmids among Escherichia coli causing urinary tract infections in Abeokuta, Nigeria. There is a high level of resistance to many antimicrobials that are frequently used in Abeokuta, Nigeria.

  4. Antimicrobial resistance in commensal Escherichia coli isolated from animals at slaughter

    Science.gov (United States)

    Wasyl, Dariusz; Hoszowski, Andrzej; Zając, Magdalena; Szulowski, Krzysztof

    2013-01-01

    Monitoring of antimicrobial resistance in commensal Escherichia coli (N = 3430) isolated from slaughtered broilers, laying hens, turkeys, swine, and cattle in Poland has been run between 2009 and 2012. Based on minimal inhibitory concentration (MIC) microbiological resistance to each of 14 tested antimicrobials was found reaching the highest values for tetracycline (43.3%), ampicillin (42.3%), and ciprofloxacin (39.0%) whereas the lowest for colistin (0.9%), cephalosporins (3.6 ÷ 3.8%), and florfenicol (3.8%). The highest prevalence of resistance was noted in broiler and turkey isolates, whereas it was rare in cattle. That finding along with resistance patterns specific to isolation source might reflect antimicrobial consumption, usage preferences or management practices in specific animals. Regression analysis has identified changes in prevalence of microbiological resistance and shifts of MIC values. Critically important fluoroquinolone resistance was worrisome in poultry isolates, but did not change over the study period. The difference (4.7%) between resistance to ciprofloxacin and nalidixic acid indicated the scale of plasmid-mediated quinolone resistance. Cephalosporin resistance were found in less than 3.8% of the isolates but an increasing trends were observed in poultry and MIC shift in the ones from cattle. Gentamycin resistance was also increasing in E. coli of turkey and cattle origin although prevalence of streptomycin resistance in laying hens decreased considerably. Simultaneously, decreasing MIC for phenicols observed in cattle and layers isolates as well as tetracycline values in E. coli from laying hens prove that antimicrobial resistance is multivariable phenomenon not only directly related to antimicrobial usage. Further studies should elucidate the scope of commensal E. coli as reservoirs of resistance genes, their spread and possible threats for human and animal health. PMID:23935596

  5. Genotyping and serotyping of macrolide and multidrug resistant Streptococcus pneumoniae isolated from carrier children

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    S F Swedan

    2016-01-01

    Full Text Available Aims: Streptococcus pneumoniae, an opportunistic pathogen commonly carried asymptomatically in the nasopharynx of children, is associated with increasing rates of treatment failures due to a worldwide increase in drug resistance. We investigated the carriage of S. pneumoniae in children 5 years or younger, the identity of prevalent serotypes, the rates of resistance to macrolides and other antimicrobial agents and the genotypes responsible for macrolide resistance. Materials and Methods: Nasopharyngeal swabs were collected from 157 children under 5 years for cultural isolation of S. pneumoniae. Antibiogram of isolates  was determined using the disk diffusion test, and the minimal inhibitory concentration to macrolides was determined using the E-test. Isolate serotypes and macrolide resistance genes, erm(B and mef(E, were identified using multiplex polymerase chain reactions. Results: S. pneumoniae was recovered from 33.8% of children; 41.9% among males and 21.9% among females (P = 0.009. The highest carriage rate occurred among age groups 7-12 months and 49-60 months. Most frequent serotypes were 19F, 6A/B, 11A, 19A, 14 and 15B/C.  Resistance to macrolides was 60.4%. Resistance to oxacillin, trimethoprim/sulfamethoxazole and clindamycin was present among 90.6%, 54.7% and 32.1% of isolates, respectively. All isolates were susceptible to chloramphenicol, levofloxacin and vancomycin. Isolates resistant to one or more macrolide drugs were more likely to be multidrug resistant. Resistance to clindamycin or oxacillin coexisted with macrolide resistance. Among the erythromycin-resistant isolates, erm(B, mef(E and erm(B and mef(E genes were present at rates of 43.8%, 37.5% and 6.3%, respectively. Erm(B and mef(E were associated with very high level and moderate-to-high level resistance to macrolides, respectively. Conclusion: A significant proportion of children harboured macrolide and multidrug-resistant S. pneumoniae.

  6. Characterization of Methicillin-Resistant Staphylococcus aureus Isolated from Healthy Turkeys and Broilers Using DNA Microarrays

    Science.gov (United States)

    El-Adawy, Hosny; Ahmed, Marwa; Hotzel, Helmut; Monecke, Stefan; Schulz, Jochen; Hartung, Joerg; Ehricht, Ralf; Neubauer, Heinrich; Hafez, Hafez M.

    2016-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is a major human health problem and recently, domestic animals are described as carriers and possible reservoirs. Twenty seven S. aureus isolates from five turkey farms (n = 18) and two broiler farms (n = 9) were obtained by culturing of choana and skin swabs from apparently healthy birds, identified by Taqman-based real-time duplex nuc-mecA-PCR and characterized by spa typing as well as by a DNA microarray based assay which covered, amongst others, a considerable number of antibiotic resistance genes, species controls, and virulence markers. The antimicrobial susceptibility profiles were tested by agar diffusion assays and genotypically confirmed by the microarray. Five different spa types (3 in turkeys and 2 in broilers) were detected. The majority of MRSA isolates (24/27) belonged to clonal complex 398-MRSA-V. The most frequently occurring spa types were accordingly t011, t034, and t899. A single CC5-MRSA-III isolated from turkey and CC398-MRSA with an unidentified/truncated SCCmec element in turkey and broiler were additionally detected. The phenotypic antimicrobial resistance profiles of S. aureus isolated from both turkeys and broilers against 14 different antimicrobials showed that all isolates were resistant to ampicillin, cefoxitin, oxacillin, doxycycline, and tetracycline. Moreover, all S. aureus isolated from broilers were resistant to erythromycin and azithromycin. All isolates were susceptible to gentamicin, chloramphenicol, sulphonamides, and fusidic acid. The resistance rate against ciprofloxacin was 55.6% in broiler isolates and 42.1% in turkey isolates. All tetracycline resistant isolates possessed genes tetK/M. All erythromycin-resistant broiler isolates carried ermA. Only one broiler isolate (11.1%) carried genes ermA, ermB, and ermC, while 55.6% of turkey isolates possessed ermA and ermB genes. Neither PVL genes (lukF/S-PV), animal-associated leukocidin (lukM and luk-P83) nor the gene encoding

  7. A survey of antibiotic resistance among E. coli strains isolated from poultry in Karachi.

    Science.gov (United States)

    Ansari, F A; Khatoon, H

    1999-01-01

    Studies were carried out to investigate the incidence of multiple antibiotic resistance among E .coli (total 152) isolated from poultry in Karachi to eight commonly used antibiotics: ampicillin (A), chloramphenicol (C), gentamycin (G), anamycin (K), neomycin (N), polymyxin B (P), streptomycin (S) and tetracycline (T) at the levels of 50 microg/ml, 100 microg/ml and 500 microg/ml. Tables of the results are given, showing the number of resistant strains of different patterns of antibiotic resistance at different levels. A comparison of antibiotic resistance to different number of antibiotics and the frequency of resistance to individual antibiotic at different levels is also reported. The highest frequency of resistance was against tetracycline whereas the lowest frequency of resistance was against gentamycin. Thirty R plasmids were isolated from the resistant strains and will be reported elsewhere.

  8. Fitness and Competitive Ability of Botrytis cinerea Isolates with Resistance to Multiple Chemical Classes of Fungicides.

    Science.gov (United States)

    Chen, S N; Luo, C X; Hu, M J; Schnabel, G

    2016-09-01

    Resistance to multiple chemical classes of fungicides in Botrytis cinerea isolates from eastern United States strawberry fields is common and strategies to control them are needed. In this study, we compared fitness and competitive ability of eight sensitive isolates (S), eight isolates resistant to five or six chemical classes of fungicides but not to phenylpyrroles (5CCR), and eight isolates resistant to six or seven chemical classes including phenylpyrroles (6CCR/MDR1h). The latter included the MDR1h phenotype due to overexpression of atrB based on Δ497V/L in mrr1. The 6CCR/MDR1h isolates grew more slowly at 4°C on potato dextrose agar, and both 5CCR and 6CCR/MDR1h isolates were hypersensitive to osmotic stress compared with S isolates. In contrast, no differences were found in oxidative sensitivity, aggressiveness, and spore production in vivo, and sclerotia production and viability in vitro. In competition experiments, the 5CCR and 6CCR/MDR1h isolates were both outcompeted by S isolates and 6CCR/MDR1h isolates were outcompeted by 5CCR isolates in the absence of fungicide pressure. Under selective pressure of a fludioxonil/pyraclostrobin rotation, the 6CCR/MDR1h isolates dominated after coinoculation with 5CCR and S isolates. The competitive disadvantage of 5CCR and especially 6CCR/MDR1h isolates suggest that, in the absence of fungicide selection pressure, S isolates may reduce inoculum potential of multifungicide-resistant isolates under field conditions.

  9. Antimicrobial resistance in clinical Escherichia coli isolates from poultry and livestock, China.

    Science.gov (United States)

    Yassin, Afrah Kamal; Gong, Jiansen; Kelly, Patrick; Lu, Guangwu; Guardabassi, Luca; Wei, Lanjing; Han, Xiangan; Qiu, Haixiang; Price, Stuart; Cheng, Darong; Wang, Chengming

    2017-01-01

    Poultry and livestock are the most important reservoirs for pathogenic Escherichia coli and use of antimicrobials in animal farming is considered the most important factor promoting the emergence, selection and dissemination of antimicrobial-resistant microorganisms. The aim of our study was to investigate antimicrobial resistance in E. coli isolated from food animals in Jiangsu, China. The disc diffusion method was used to determine susceptibility to 18 antimicrobial agents in 862 clinical isolates collected from chickens, ducks, pigs, and cows between 2004 and 2012. Overall, 94% of the isolates showed resistance to at least one drug with 83% being resistance to at least three different classes of antimicrobials. The isolates from the different species were most commonly resistant to tetracycline, nalidixic acid, sulfamethoxazole, trimethoprim/sulfamethoxazole and ampicillin, and showed increasing resistance to amikacin, aztreonam, ceftazidime, cefotaxime, chloramphenicol, ciprofloxacin. They were least resistant to amoxicillin/clavulanic acid (3.4%) and ertapenem (0.2%). MDR was most common in isolates from ducks (44/44, 100%), followed by chickens (568/644, 88.2%), pigs (93/113, 82.3%) and cows (13/61, 21.3%). Our finding that clinical E. coli isolates from poultry and livestock are commonly resistant to multiple antibiotics should alert public health and veterinary authorities to limit and rationalize antimicrobial use in China.

  10. Comparative RNA-seq-Based Transcriptome Analysis of the Virulence Characteristics of Methicillin-Resistant and -Susceptible Staphylococcus pseudintermedius Strains Isolated from Small Animals.

    Science.gov (United States)

    Couto, Natacha; Belas, Adriana; Oliveira, Manuela; Almeida, Paulo; Clemente, Carla; Pomba, Constança

    2016-02-01

    Staphylococcus pseudintermedius is often associated with pyoderma, which can turn into a life-threatening disease. The dissemination of highly resistant isolates has occurred in the last 10 years and has challenged antimicrobial treatment of these infections considerably. We have compared the carriage of virulence genes and biofilm formation between methicillin-resistant and methicillin-susceptible S. pseudintermedius (MRSP and MSSP, respectively) isolates and their in vitro gene expression profiles by transcriptome sequencing (RNA-seq). Isolates were relatively unevenly distributed among the four agr groups, and agr type III predominated in MRSP. Five virulence genes were detected in all isolates. Only the spsO gene was significantly associated with MSSP isolates (P = 0.04). All isolates produced biofilm in brain heart infusion broth (BHIB)-4% NaCl. MSSP isolates produced more biofilm on BHIB and BHIB-1% glucose media than MRSP isolates (P = 0.03 and P = 0.02, respectively). Virulence genes encoding surface proteins and toxins (spsA, spsB, spsD, spsK, spsL, spsN, nucC, coa, and luk-I) and also prophage genes (encoding phage capsid protein, phage infection protein, two phage portal proteins, and a phage-like protein) were highly expressed in the MRSP isolate (compared with the MSSP isolate), suggesting they may play a role in the rapid and widespread dissemination of MRSP. This study indicates that MRSP may upregulate surface proteins, which may increase the adherence of MRSP isolates (especially sequence type 71 [ST71]) to corneocytes. MSSP isolates may have an increased ability to form biofilm under acidic circumstances, through upregulation of the entire arc operon. Complete understanding of S. pseudintermedius pathogenesis and host-pathogen signal interaction during infections is critical for the treatment and prevention of S. pseudintermedius infections.

  11. Multidrug resistant Escherichia coli strains isolated from urine sample, University of Gondar Hospital, Northwest Ethiopia

    Institute of Scientific and Technical Information of China (English)

    Setegn Eshetie; Fentahun Tarekegn; Gemechu Kumera; Feleke Mekonnen

    2016-01-01

    Objective: To assess multidrug resistant (MDR) Escherichia coli (E. coli) isolates from patients with urinary tract infection. Methods: From February to June 2014, a cross sectional study was conducted among urinary tract infection patients at the University of Gondar Hospital. Culture and disk diffusion method were used for E. coli isolation and to determine the antibiotic susceptibility patterns. Data were entered and analyzed using SPSS version 20. P Results: A total of 112 E. coli isolates were identified and the rate of isolation was higher among female participants (28.7%; P = 0.03). Of the isolates, 104 (92.9%) were MDR E. coli; and the isolates showed high resistance rates towards ampicillin (99%), cotrimoxazole (69%), chloramphenicol (58.7%), gentamycin (56.7%) and ceftazidime (55.8%). However, comparative isolates showed low resistance rates to ciprofloxacin (1%), cefepime (8.7%), and ceftriaxone (11.5%). Moreover, resistance rates of MDR E. coli isolates were significantly higher than non-MDR strains for ceftazidime (55.8% versus 12.5%; P = 0.015), and ampicillin (99% versus 87.5%; P = 0.018). Conclusions: High prevalence of MDR E. coli isolates was observed in this study. Regular monitoring of antibiotic resistance rates is necessarily required to improve and revise empirical antibiotic therapy protocols.

  12. Integron mediated multidrug resistance in extended spectrum beta-lactamase producing clinical isolates of Klebsiella pneumoniae

    Directory of Open Access Journals (Sweden)

    Maryam Mobarak-Qamsari

    2013-09-01

    Full Text Available The present study describes integron mediated multiple antibiotic resistance in extended-spectrum β-lactamase producing clinical isolates of Klebsiella pneumoniae. One hundred and four clinical isolates of K. pneumoniae from two Iranian hospitals were screened for extended-spectrum β-lactamase production and susceptibility of the extended-spectrum β-lactamase producing isolates was determined to 17 antibiotics by disc diffusion. Presence of integron classes 1, 2 and 3 was detected by PCR and integrase specific primers. Isolates harboring class 1 integron were then screened for variable regions using PCR. Fifty isolates (48% produced extended-spectrum β-lactamases among which, 22 (44% harbored class 1, 3 (6% carried class 2 and none contained class 3 integons. Integron carriage was significantly associated with higher rates of multiple antibiotic resistance in extended-spectrum β-lactamase producing clinical isolates of K. pneumoniae. Integron harboring isolates were more resistant to aztreonam (51.3%, ceftazidime (42.6%, cefotaxime (43.3%, cefepime (24.6%, kanamycin (43.2%, tobramycin (30.7%, norfloxcacin (32% and spectinomycin (25.6% compared to the organisms without integrons. On the other hand, resistance to nitrofurantoin and streptomycin was significantly higher among the integron negative isolates. PCR amplification of class1 integron variable regions revealed 9 different sized DNA fragments and isolates with similar profiles for class 1 integron variable regions showed the same antibiotic resistance phenotypes.

  13. Integron mediated multidrug resistance in extended spectrum beta-lactamase producing clinical isolates of Klebsiella pneumoniae

    Science.gov (United States)

    Mobarak-Qamsari, Maryam; Ashayeri-Panah, Mitra; Eftekhar, Freshteh; Feizabadi, Mohammad Mehdi

    2013-01-01

    The present study describes integron mediated multiple antibiotic resistance in extended-spectrum β-lactamase producing clinical isolates of Klebsiella pneumoniae. One hundred and four clinical isolates of K. pneumoniae from two Iranian hospitals were screened for extended-spectrum β-lactamase production and susceptibility of the extended-spectrum β-lactamase producing isolates was determined to 17 antibiotics by disc diffusion. Presence of integron classes 1, 2 and 3 was detected by PCR and integrase specific primers. Isolates harboring class 1 integron were then screened for variable regions using PCR. Fifty isolates (48%) produced extended-spectrum β-lactamases among which, 22 (44%) harbored class 1, 3 (6%) carried class 2 and none contained class 3 integons. Integron carriage was significantly associated with higher rates of multiple antibiotic resistance in extended-spectrum β-lactamase producing clinical isolates of K. pneumoniae. Integron harboring isolates were more resistant to aztreonam (51.3%), ceftazidime (42.6%), cefotaxime (43.3%), cefepime (24.6%), kanamycin (43.2%), tobramycin (30.7%), norfloxcacin (32%) and spectinomycin (25.6%) compared to the organisms without integrons. On the other hand, resistance to nitrofurantoin and streptomycin was significantly higher among the integron negative isolates. PCR amplification of class1 integron variable regions revealed 9 different sized DNA fragments and isolates with similar profiles for class 1 integron variable regions showed the same antibiotic resistance phenotypes. PMID:24516451

  14. Isolation and partial characterization of actinomycetes with antimicrobial activity against multidrug resistant bacteria

    Institute of Scientific and Technical Information of China (English)

    Smriti Singh; Pramod Kumar; N Gopalan; Bhuvnesh Shrivastava; RC Kuhad; Hotam Singh Chaudhary

    2012-01-01

    Objective: To isolate strains of Actinomycetes from different locations of Gwalior to evaluate its antimicrobial activity against multidrug resistant pathogenic strains. Method: Soil samples collected from different niche habitats of Gwalior were serially diluted and plated on selective media. Potential colonies were further purified and stored in agar slants and glycerol stocks. Isolates were biochemically characterized and purified isolates were test against pathogenic microorganisms for screening. Isolates with antagonistic properties were inoculated in production media and secondary metabolites or antimicrobial products were extracted. Result: The seven actinomycetes strains showing maximum antibacterial activity were isolated further characterized based on their colony characteristics and biochemical analyses. The isolates were screened for their secondary metabolites activity on three human pathogenic bacteria are Escherichia coli (E. coli), Methicillin-Resistant Staphylococcus aureus (S. aureus) and Vancomycin-Resistant Enterococci (VRE). Discussion: The strain MITS 1005 was found to be more active against the test bacteria.

  15. Detection of plasmid-mediated IMP-1 metallo-β-lactamase and quinolone resistance determinants in an ertapenem-resistant Enterobacter cloacae isolate

    Institute of Scientific and Technical Information of China (English)

    Li-rong CHEN; Hong-wei ZHOU; Jia-chang CAI; Rong ZHANG; Gong-xiang CHEN

    2009-01-01

    Objective: To investigate the mechanism of carbapenem resistance and the occurrence of plasmid-mediated quinolone resistance determinants qnr and aac(6')-Ib-cr in a clinical isolate of Enterobacter cloacae. Methods: An ertapenem-resistant E. cloacae ZY106, which was isolated from liquor puris of a female gastric cancer patient in a Chinese hospital, was investigated. Antibiotic susceptibilities were determined by agar dilution method. Conjugation experiments, isoelectric focusing, polymerase chain reaction (PCR), and DNA sequence analyses of plasmid-mediated carbapenemases and quinolone resistance determinants were preformed to confirm the genotype. Outer membrane proteins (OMPs) were examined by urea-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Urea-SDS-PAGE). Results: Minimum inhibitory concentrations (MCs) of imipenem, mer-openem, and ertapenem for ZY106 were 2,4, and 16 ug/ml, respectively. Conjugation studies with Escherichia coli resulted in the transfer of significantly reduced carbapenem susceptibility. ZY106 produced IMP-1 metallo-p-lactamase and CTX-M-3 extended-spectrum P-lactamase, and E. coli transconjugant produced IMP-1. Plasmid-mediated quinolone resistance determinant qnrSI was detected in ZY106. Transfer of the qnrSI-encoding-plasmid into E. coli by conjugation resulted in intermediate resistance to ciprofloxacin in E. coli transconjugant. Urea-SDS-PAGE analysis of OMPs showed that ZY106 lacked an OMP of approximately 38 KDa. Conclusion: It is the first IMP-1-producing Enterobacteriaceae in China and the first report of a clinical isolate that harbors both blaIMP and qnrS genes as well. The blaIMP-1, blaCTX-M-3, and qnrSl are encoded at three different plasmids. IMP-1 combined with the loss of an OMP possibly resulted in ertapenem resistance and reduced imipenem and mero-penem susceptibility in E. cloacae.

  16. Bacteriophages Isolated from Chicken Meat and the Horizontal Transfer of Antimicrobial Resistance Genes

    Science.gov (United States)

    Shousha, Amira; Awaiwanont, Nattakarn; Sofka, Dmitrij; Smulders, Frans J. M.; Paulsen, Peter; Szostak, Michael P.; Humphrey, Tom

    2015-01-01

    Antimicrobial resistance in microbes poses a global and increasing threat to public health. The horizontal transfer of antimicrobial resistance genes was thought to be due largely to conjugative plasmids or transposons, with only a minor part being played by transduction through bacteriophages. However, whole-genome sequencing has recently shown that the latter mechanism could be highly important in the exchange of antimicrobial resistance genes between microorganisms and environments. The transfer of antimicrobial resistance genes by phages could underlie the origin of resistant bacteria found in food. We show that chicken meat carries a number of phages capable of transferring antimicrobial resistance. Of 243 phages randomly isolated from chicken meat, about a quarter (24.7%) were able to transduce resistance to one or more of the five antimicrobials tested into Escherichia coli ATCC 13706 (DSM 12242). Resistance to kanamycin was transduced the most often, followed by that to chloramphenicol, with four phages transducing tetracycline resistance and three transducing ampicillin resistance. Phages able to transduce antimicrobial resistance were isolated from 44% of the samples of chicken meat that we tested. The statistically significant (P = 0.01) relationship between the presence of phages transducing kanamycin resistance and E. coli isolates resistant to this antibiotic suggests that transduction may be an important mechanism for transferring kanamycin resistance to E. coli. It appears that the transduction of resistance to certain antimicrobials, e.g., kanamycin, not only is widely distributed in E. coli isolates found on meat but also could represent a major mechanism for resistance transfer. The result is of high importance for animal and human health. PMID:25934615

  17. Emergence of a Streptococcus pneumoniae clinical isolate highly resistant to telithromycin and fluoroquinolones.

    Science.gov (United States)

    Faccone, Diego; Andres, Patricia; Galas, Marcelo; Tokumoto, Marta; Rosato, Adriana; Corso, Alejandra

    2005-11-01

    Streptococcus pneumoniae is a major pathogen causing community-acquired pneumonia and acute bronchitis. Macrolides, fluoroquinolones (FQs), and, recently, telithromycin (TEL) constitute primary therapeutic options, and rare cases of resistance have been reported. In this report, we describe the emergence of an S. pneumoniae clinical isolate with high-level TEL resistance (MIC, 256 microg/ml) and simultaneous resistance to FQs. Ongoing studies are oriented to elucidate the precise mechanism of resistance to TEL.

  18. Improved Detection by Next-Generation Sequencing of Pyrazinamide Resistance in Mycobacterium tuberculosis Isolates

    OpenAIRE

    Maningi, Nontuthuko E.; Daum, Luke T.; Rodriguez, John D.; Mphahlele, Matsie; Peters, Remco P. H.; Fischer, Gerald W.; Chambers, James P.; Fourie, P. Bernard

    2015-01-01

    The technical limitations of common tests used for detecting pyrazinamide (PZA) resistance in Mycobacterium tuberculosis isolates pose challenges for comprehensive and accurate descriptions of drug resistance in patients with multidrug-resistant tuberculosis (MDR-TB). In this study, a 606-bp fragment (comprising the pncA coding region plus the promoter) was sequenced using Ion Torrent next-generation sequencing (NGS) to detect associated PZA resistance mutations in 88 recultured MDR-TB isolat...

  19. Bacteriophages Isolated from Chicken Meat and the Horizontal Transfer of Antimicrobial Resistance Genes.

    Science.gov (United States)

    Shousha, Amira; Awaiwanont, Nattakarn; Sofka, Dmitrij; Smulders, Frans J M; Paulsen, Peter; Szostak, Michael P; Humphrey, Tom; Hilbert, Friederike

    2015-07-01

    Antimicrobial resistance in microbes poses a global and increasing threat to public health. The horizontal transfer of antimicrobial resistance genes was thought to be due largely to conjugative plasmids or transposons, with only a minor part being played by transduction through bacteriophages. However, whole-genome sequencing has recently shown that the latter mechanism could be highly important in the exchange of antimicrobial resistance genes between microorganisms and environments. The transfer of antimicrobial resistance genes by phages could underlie the origin of resistant bacteria found in food. We show that chicken meat carries a number of phages capable of transferring antimicrobial resistance. Of 243 phages randomly isolated from chicken meat, about a quarter (24.7%) were able to transduce resistance to one or more of the five antimicrobials tested into Escherichia coli ATCC 13706 (DSM 12242). Resistance to kanamycin was transduced the most often, followed by that to chloramphenicol, with four phages transducing tetracycline resistance and three transducing ampicillin resistance. Phages able to transduce antimicrobial resistance were isolated from 44% of the samples of chicken meat that we tested. The statistically significant (P = 0.01) relationship between the presence of phages transducing kanamycin resistance and E. coli isolates resistant to this antibiotic suggests that transduction may be an important mechanism for transferring kanamycin resistance to E. coli. It appears that the transduction of resistance to certain antimicrobials, e.g., kanamycin, not only is widely distributed in E. coli isolates found on meat but also could represent a major mechanism for resistance transfer. The result is of high importance for animal and human health.

  20. Echinoderms from Azores islands: an unexpected source of antibiotic resistant Enterococcus spp. and Escherichia coli isolates.

    Science.gov (United States)

    Marinho, Catarina; Silva, Nuno; Pombo, Sofia; Santos, Tiago; Monteiro, Ricardo; Gonçalves, Alexandre; Micael, Joana; Rodrigues, Pedro; Costa, Ana Cristina; Igrejas, Gilberto; Poeta, Patrícia

    2013-04-15

    The prevalence of antibiotic resistance and the implicated mechanisms of resistance were evaluated in Enterococcus spp. and Escherichia coli, isolated from a total of 250 faecal samples of echinoderms collected from Azorean waters (Portugal). A total of 144 enterococci (120 Enterococcus faecium, 14 E. hirae, 8 E. faecalis, 2 E. gallinarum) and 10 E. coli were recovered. High percentages of resistance in enterococci were found for erythromycin, ampicillin, tetracyclin and ciprofloxacin. The erm(A) or erm(B), tet(M) and/or tet(L), vat(D), aac(6')-aph(2″) and aph(3')-IIIa genes were found in isolates resistant to erythromycin, tetracycline, quinupristin/dalfopristin, high-level gentamicin and high-level kanamycin, respectively. Resistance in E. coli isolates was detected for streptomycin, amikacin, tetracycline and tobramycin. The aadA gene was found in streptomycin-resistant isolates and tet(A)+tet(B) genes in tetracycline-resistant isolates. The data recovered are essential to improve knowledge about the dissemination of resistant strains through marine ecosystems and the possible implications involved in transferring these resistances either to other animals or to humans.

  1. Protein phosphorylation in isolated hepatocytes of septic and endotoxemic rats

    Energy Technology Data Exchange (ETDEWEB)

    Deaciuc, I.V.; Spitzer, J.A. (Louisiana State Univ. Medical Center, New Orleans (USA))

    1989-11-01

    The purpose of this study was to investigate possible alterations induced by sepsis and endotoxicosis in the late phase of Ca2+-dependent signaling in rat liver. Hepatocytes isolated from septic or chronically endotoxin (ET)-treated rats were labeled with (32P)H3PO4 and stimulated with various agents. Proteins were resolved by one-dimensional polyacrylamide gel electrophoresis and autoradiographed. Vasopressin (VP)- and phenylephrine (PE)-induced responses were attenuated in both septic and ET-treated rats for cytosolic and membrane proteins compared with their respective controls. Glucagon and 12-O-myristate phorbol-13-acetate (TPA) affected only the phosphorylation of membrane proteins. Glucagon-induced changes in the phosphorylation of membrane proteins were affected by both sepsis and endotoxicosis, whereas TPA-stimulated phosphorylation was lowered only in endotoxicosis. Response to the Ca2+ ionophore A23187 was depressed in septic rats for cytosolic proteins. The phosphorylation of two cytosolic proteins, i.e., 93 and 61 kDa (previously identified as glycogen phosphorylase and pyruvate kinase, respectively), in response to VP, PE, and A23187 was severely impaired by endotoxicosis and sepsis. TPA did not affect the phosphorylation state of these two proteins. The results show that sepsis and endotoxicosis produce perturbations of the phosphorylation step in Ca2+ transmembrane signaling. Such changes can explain alterations of glycogenolysis and gluconeogenesis associated with sepsis and endotoxicosis.

  2. Protein expression on Cr resistant microorganism using electrophoresis method

    Directory of Open Access Journals (Sweden)

    SAJIDAN

    2009-01-01

    Full Text Available Fatmawati U, Suranto, Sajidan. 2009. Protein expression on Cr resistant microorganism using electrophoresis method. Nusantara Bioscience 1: 31-37. Hexavalent chromium (Cr(VI is known as toxic heavy metals, so the need is reduced to Cr(III is much less toxicity. Pseudomonas aeruginosa, Pseudomonas putida, Klebsiella pneumoniae, Pantoea sp. and Saccharomyces cerevisiae are resistant Cr(VI microorganism and have ability to reduce Cr(VI. The aim of this research is to know ability of microorganism to reduce Cr(VI and to know protein band pattern between Cr(VI resistant microorganism and non resistant microorganism which inoculated on LB broth. SDS-PAGE was used to indentify protein expression. While, Cr(VI concentration was identified by 1.5 diphenylcarbazide method. The quantitative data was analyzed by two factorial ANOVA that continued with DMRT at 1% level test. The qualitative data i.e. protein expression analyzed by relative mobility (Rf. The results showed that the ability of microorganisms to reduce Cr(VI at initial concentration of 0.5 ppm, 1 ppm, 5 ppm and 10 ppm may vary, the average percentage of the ability of each microorganism in reducing Cr(VI is P. putida (65% > S. cerevisiae (64.45% >. P. aeruginosa (60.73% > Pantoea sp. (50.22% > K. pneumoniae (47.82% > without microorganisms (34.25%. The adding microorganisms have significantly influenced toward reduction of Cr(VI. The SDS-PAGE shows that protein expression between resistant and not resistant microorganisms are no different, but resistant microorganisms have more protein (protein band is thicker.

  3. Molecular surveillance of drug resistance through imported isolates of Plasmodium falciparum in Europe

    DEFF Research Database (Denmark)

    Jelinek, Tomas; Peyerl-Hoffmann, Gabriele; Mühlberger, Nikolai;

    2002-01-01

    BACKGROUND: Results from numerous studies point convincingly to correlations between mutations at selected genes and phenotypic resistance to antimalarials in Plasmodium falciparum isolates. In order to move molecular assays for point mutations on resistance-related genes into the realm of applied...... tools for surveillance, we investigated a selection of P. falciparum isolates that were imported during the year 2001 into Europe to study the prevalence of resistance-associated point mutations at relevant codons. In particular, we tested for parasites which were developing resistance to antifolates...... and chloroquine. The screening results were used to map the prevalence of mutations and, thus, levels of potential drug resistance in endemic areas world-wide. RESULTS: 337 isolates have been tested so far. Prevalence of mutations that are associated with resistance to chloroquine on the pfcrt and pfmdr genes...

  4. In Vitro Activity of ASP2397 against Aspergillus Isolates with or without Acquired Azole Resistance Mechanisms.

    Science.gov (United States)

    Arendrup, Maiken Cavling; Jensen, Rasmus Hare; Cuenca-Estrella, Manuel

    2015-11-09

    ASP2397 is a new compound with a novel and as-yet-unknown target different from that of licensed antifungal agents. It has activity against Aspergillus and Candida glabrata. We compared its in vitro activity against wild-type and azole-resistant A. fumigatus and A. terreus isolates with that of amphotericin B, itraconazole, posaconazole, and voriconazole. Thirty-four isolates, including 4 wild-type A. fumigatus isolates, 24 A. fumigatus isolates with alterations in CYP51A TR/L98H (5 isolates), M220 (9 isolates), G54 (9 isolates), and HapE (1 isolate), and A. terreus isolates (2 wild-type isolates and 1 isolate with an M217I CYP51A alteration), were analyzed. EUCAST E.Def 9.2 and CLSI M38-A2 MIC susceptibility testing was performed. ASP2397 MIC50 values (in milligrams per liter, with MIC ranges in parentheses) determined by EUCAST and CLSI were 0.5 (0.25 to 1) and 0.25 (0.06 to 0.25) against A. fumigatus CYP51A wild-type isolates and were similarly 0.5 (0.125 to >4) and 0.125 (0.06 to >4) against azole-resistant A. fumigatus isolates, respectively. These values were comparable to those for amphotericin B, which were 0.25 (0.125 to 0.5) and 0.25 (0.125 to 0.25) against wild-type isolates and 0.25 (0.125 to 1) and 0.25 (0.125 to 1) against isolates with azole resistance mechanisms, respectively. In contrast, MICs for the azole compounds were elevated and highest for itraconazole: >4 (1 to >4) and 4 (0.5 to >4) against isolates with azole resistance mechanisms compared to 0.125 (0.125 to 0.25) and 0.125 (0.06 to 0.25) against wild-type isolates, respectively. ASP2397 was active against A. terreus CYP51A wild-type isolates (MIC 0.5 to 1), whereas MICs of both azole and ASP2397 were elevated for the mutant isolate. ASP2397 displayed in vitro activity against A. fumigatus and A. terreus isolates which was independent of the presence or absence of azole target gene resistance mutations in A. fumigatus. The findings are promising at a time when azole-resistant A. fumigatus

  5. Anti Microbial Resistance Profile of E. coli isolates From Tropical Free Range Chickens

    Directory of Open Access Journals (Sweden)

    Okoli CI

    2005-12-01

    Full Text Available Normal intestinal flora of humans and animals constitute enormous reservoir of resistance genes for potentially pathogenic bacteria and may serve as major indictors of selection pressure exerted by anti-microbial use in a given population. A study was conducted in September 2003 at 3 purposively selected peri-urban sites spread across 3 senatorial zones of Imo state, Nigeria to determines the anti-microbial resistance profile of commensal E. coli isolated from free range chickens. The isolates were screened for anti-microbial resistance profile against 10 antibiotics using the disc diffusion method. E. coli strains from local fowls, recorded 100 and 78.9% resistances against ampicillin and cotrimoxazole respectively, while resistance rates against ciprofloxacin, gentamycin and norfloxacin were 0.0, 5.3 and 5.3% respectively. Isolates from free-range cockerels recorded 100% resistance against norfloxacin, cotrimoxazole and ampicillin and 83.3% against nitrofurantoin. Similarly, isolates from old layers, recorded 100% resistance against nitrofurantoin, cotrimoxazole, tetracycline, chloramphenicol and ampicillin and 80% against nalidixic acid. Similar resistance trends were observed in E. coli isolates from free-range turkeys and broiler roosters against the inexpensive broad-spectrum first line antibiotics (ampicillin, nalidixic acid, cotrimoxazole, nitrofurantoin and chloramphenicol, although values varied slightly across poultry types. Resistances against gentamycin were consistently low in isolates from the different types of poultry. Twenty-nine resistance patterns were observed in the E. coli isolates with predominant patterns being distributed widely across poultry types indicating a striking diversity of resistance patterns in the areas.

  6. Phage types and antimicrobial resistance among Danish bovine Staphylococcus aureus isolates since the 1950s

    DEFF Research Database (Denmark)

    Vintov, Jan; Aarestrup, Frank Møller; Zinn, C. E.

    2003-01-01

    A total of 292 bovine Staphylococcus aureus isolates obtained from the 1950s (86 isolates), 1992 (107 isolates), and 2000 (99 isolates) were examined for antimicrobial susceptibility and phage typing. The same types of S. aureus (80, 52, 3A, 3A/3C, 42E, 77) were found among the isolates from all...... three time periods, representing 43.3% of the typeable isolates. This indicates that the Danish S. aureus population related to bovine mastitis has remained relatively unchanged over the last 50 years. The occurrence of antimicrobial resistance has remained low in Denmark in comparison to other...

  7. Emergence of clonally related multidrug resistant Haemophilus influenzae with penicillin-binding protein 3-mediated resistance to extended-spectrum cephalosporins, Norway, 2006 to 2013.

    Science.gov (United States)

    Skaare, D; Anthonisen, I L; Kahlmeter, G; Matuschek, E; Natås, O B; Steinbakk, M; Sundsfjord, A; Kristiansen, B E

    2014-12-11

    Resistance to cephalosporins in Haemophilus influenzae is usually caused by characteristic alterations in penicillin-binding protein 3 (PBP3), encoded by the ftsI gene. Resistance to extended-spectrum cephalosporins is associated with high-level PBP3-mediated resistance (high-rPBP3), defined by the second stage S385T substitution in addition to a first stage substitution (R517H or N526K). The third stage L389F substitution is present in some high-rPBP3 strains. High-rPBP3 H. influenzae are considered rare outside Japan and Korea. In this study, 30 high-rPBP3 isolates from Norway, collected between 2006 and 2013, were examined by serotyping, multilocus sequence typing (MLST), ftsI sequencing, detection of beta-lactamase genes and minimum inhibitory concentration (MIC) determination. MICs were interpreted according to clinical breakpoints from the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Respiratory isolates predominated (proportion: 24/30). The 30 isolates included one serotype f isolate, while the remaining 29 lacked polysaccharide capsule genes. Resistance to extended-spectrum cephalosporins (cefixime, 29 isolates/30 isolates; cefepime, 28/30; cefotaxime, 26 /30; ceftaroline, 26/30; ceftriaxone, 14/30), beta-lactamase production (11/30) and co-resistance to non-beta-lactams (trimethoprim-sulfamethoxazole, 13/30; tetracycline, 4/30; chloramphenicol, 4/30; ciprofloxacin, 3/30) was frequent. The N526K substitution in PBP3 was present in 23 of 30 isolates; these included a blood isolate which represents the first invasive S385T + N526K isolate reported from Europe. The L389F substitution, present in 16 of 30 isolates, coincided with higher beta-lactam MICs. Non-susceptibility to meropenem was frequent in S385T + L389F + N526K isolates (8/12). All 11 beta-lactamase positive isolates were TEM-1. Five clonal groups of two to 10 isolates with identical MLST-ftsI allelic profiles were observed, including the first reported high-rPBP3

  8. Heterologously expressed bacterial and human multidrug resistance proteins confer cadmium resistance to Escherichia coli

    NARCIS (Netherlands)

    Achard-Joris, M; van Saparoea, HBV; Driessen, AJM; Bourdineaud, JP; Bourdineaud, Jean-Paul

    2005-01-01

    The human MDR1 gene is induced by cadmium exposure although no resistance to this metal is observed in human cells overexpressing hMDR1. To access the role of MDR proteins in cadmium resistance, human MDR1, Lactococcus lactis lmrA, and Oenococcus oeni omrA were expressed in an Escherichia coli tolC

  9. Primary antibiotic resistance and associated mechanisms in Helicobacter pylori isolates from Senegalese patients

    Directory of Open Access Journals (Sweden)

    Seck Abdoulaye

    2013-01-01

    Full Text Available Abstract Background Antibiotic combination therapy for Helicobacter pylori eradication must be adapted to local resistance patterns, but the epidemiology of H. pylori resistance to antibiotics is poorly documented in Africa. The aim was to determine the antibiotic resistance rates, as well as the associated molecular mechanisms, of strains isolated in Dakar, Senegal. Methods One hundred and eight H. pylori strains were isolated between 2007 and 2009 from 108 patients presenting with upper abdominal pain to the Gastroenterology Department of Le Dantec Hospital. Antimicrobial susceptibility testing was performed for amoxicillin, clarithromycin, metronidazole, levofloxacin and tetracyclin using the E-test method. Mutations in the 23S rRNA gene of clarithromycin-resistant strains and in gyrA and gyrB of levofloxacin-resistant strains were investigated. Results Isolates were characterized by no resistance to amoxicillin (0%, tetracycline (0%, and very low rate of resistance to clarithromycin (1%, but a high rate of resistance to metronidazole (85%. The clarithromycin-resistant strain displayed the A2143G mutation. A worrying rate of levofloxacin resistance was detected (15%. N87I and D91N were the most common mutations in the quinolone-resistance-determining region of gyrA. Conclusions The first-line empirical regimen for H. pylori eradication in Senegal should include clarithromycin. Increasing rates of fluoroquinolone resistance detected should discourage the use of levofloxacin-containing regimens without prior antimicrobial susceptibility testing.

  10. Dietary protein to maximize resistance training: a review and examination of protein spread and change theories

    Directory of Open Access Journals (Sweden)

    Bosse John D

    2012-09-01

    Full Text Available Abstract An appreciable volume of human clinical data supports increased dietary protein for greater gains from resistance training, but not all findings are in agreement. We recently proposed “protein spread theory” and “protein change theory” in an effort to explain discrepancies in the response to increased dietary protein in weight management interventions. The present review aimed to extend “protein spread theory” and “protein change theory” to studies examining the effects of protein on resistance training induced muscle and strength gains. Protein spread theory proposed that there must have been a sufficient spread or % difference in g/kg/day protein intake between groups during a protein intervention to see muscle and strength differences. Protein change theory postulated that for the higher protein group, there must be a sufficient change from baseline g/kg/day protein intake to during study g/kg/day protein intake to see muscle and strength benefits. Seventeen studies met inclusion criteria. In studies where a higher protein intervention was deemed successful there was, on average, a 66.1% g/kg/day between group intake spread versus a 10.2% g/kg/day spread in studies where a higher protein diet was no more effective than control. The average change in habitual protein intake in studies showing higher protein to be more effective than control was +59.5% compared to +6.5% when additional protein was no more effective than control. The magnitudes of difference between the mean spreads and changes of the present review are similar to our previous review on these theories in a weight management context. Providing sufficient deviation from habitual intake appears to be an important factor in determining the success of additional protein in enhancing muscle and strength gains from resistance training. An increase in dietary protein favorably effects muscle and strength during resistance training.

  11. Dietary protein to maximize resistance training: a review and examination of protein spread and change theories.

    Science.gov (United States)

    Bosse, John D; Dixon, Brian M

    2012-09-08

    An appreciable volume of human clinical data supports increased dietary protein for greater gains from resistance training, but not all findings are in agreement. We recently proposed "protein spread theory" and "protein change theory" in an effort to explain discrepancies in the response to increased dietary protein in weight management interventions. The present review aimed to extend "protein spread theory" and "protein change theory" to studies examining the effects of protein on resistance training induced muscle and strength gains. Protein spread theory proposed that there must have been a sufficient spread or % difference in g/kg/day protein intake between groups during a protein intervention to see muscle and strength differences. Protein change theory postulated that for the higher protein group, there must be a sufficient change from baseline g/kg/day protein intake to during study g/kg/day protein intake to see muscle and strength benefits. Seventeen studies met inclusion criteria. In studies where a higher protein intervention was deemed successful there was, on average, a 66.1% g/kg/day between group intake spread versus a 10.2% g/kg/day spread in studies where a higher protein diet was no more effective than control. The average change in habitual protein intake in studies showing higher protein to be more effective than control was +59.5% compared to +6.5% when additional protein was no more effective than control. The magnitudes of difference between the mean spreads and changes of the present review are similar to our previous review on these theories in a weight management context. Providing sufficient deviation from habitual intake appears to be an important factor in determining the success of additional protein in enhancing muscle and strength gains from resistance training. An increase in dietary protein favorably effects muscle and strength during resistance training.

  12. Isolation and Purification of Thiamine Binding Protein from Mung Bean

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    MOHAMAD SADIKIN

    2013-03-01

    Full Text Available Thiamine has fundamental role in energy metabolism. The organs mostly sensitive to the lack of thiamine levels in the body are the nervous system and the heart. Thiamine deficiency causes symptoms of polyneuritis and cardiovascular diseases. Because of its importance in the metabolism of carbohydrates, we need to measure the levels of thiamine in the body fluids by using an easy and inexpensive way without compromising the sensitivity and selectivity. An option to it is thiamine measurement based on the principle of which is analogous to ELISA, in which a thiamine binding protein (TBP act by replacing antibodies. The presence of TBP in several seeds have been reported by previous researchers, but the presence of TBP in mung beans has not been studied. This study was aimed to isolate and purify TBP from mung bean. The protein was isolated from mung bean through salting out by ammonium sulphate of 40, 70, and 90% (w/v. TBP has a negative charge as shown by cellulose acetate electrophoresis. The result obtained after salting out by ammonium sulphate was further purified bymeans of DEAE-cellulose chromatography and affinity chromatography. In precipitation of 90% of salting out method, one peak protein was obtained by using affinity chromatography. The protein was analyzed by SDS PAGE electrophoresis. The result of SDS PAGE electrophoresis showed that TBP has a molecular weight of 72.63 kDa.

  13. COMPARATIVE PRODUCTION OF SINGLE CELL PROTEIN FROM FISH PROTEIN ISOLATE WASTAGE AND ULTRA FILTERED CHEESE WHEY

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    Soroush Haghighi-Manesh

    2013-02-01

    Full Text Available Fish protein isolate wastage and ultra filtered cheese whey were used as substrates for fermentation by Kluyveromyces marxianus to produce single cell protein, under batch and aerobic condition in which pH and temperature were adjusted to 4.5 and 35°C. The produced biomass was analyzed for protein content in different periods of time during fermentation. About 82% and 75% of total protein was produced in the first 18 h of 96 h fermentation of ultra filtered cheese whey and protein isolate wastage respectively, which can be an indication of the exponential phase of the yeast growth. The results of biomass yield measurements during 96 h process also confirm this finding. Moreover, since ultra filtered cheese whey was higher in single cell protein yield, solubility, water holding capacity, water absorption and power of biological and chemical oxygen demand reduction, and also was lower in foam overrun and stability than fish protein isolate wastage, it was selected as the suitable substrate for single cell protein production.

  14. Arsenic resistance and prevalence of arsenic resistance genes in Campylobacter jejuni and Campylobacter coli isolated from retail meats.

    Science.gov (United States)

    Noormohamed, Aneesa; Fakhr, Mohamed K

    2013-08-07

    Studies that investigate arsenic resistance in the foodborne bacterium Campylobacter are limited. A total of 552 Campylobacter isolates (281 Campylobacter jejuni and 271 Campylobacter coli) isolated from retail meat samples were subjected to arsenic resistance profiling using the following arsenic compounds: arsanilic acid (4-2,048 μg/mL), roxarsone (4-2048 μg/mL), arsenate (16-8,192 μg/mL) and arsenite (4-2,048 μg/mL). A total of 223 of these isolates (114 Campylobacter jejuni and 109 Campylobacter coli) were further analyzed for the presence of five arsenic resistance genes (arsP, arsR, arsC, acr3, and arsB) by PCR. Most of the 552 Campylobacter isolates were able to survive at higher concentrations of arsanilic acid (512-2,048 μg/mL), roxarsone (512-2,048 μg/mL), and arsenate (128-1,024 μg/mL), but at lower concentrations for arsenite (4-16 μg/mL). Ninety seven percent of the isolates tested by PCR showed the presence of arsP and arsR genes. While 95% of the Campylobacter coli isolates contained a larger arsenic resistance operon that has all of the four genes (arsP, arsR, arsC and acr3), 85% of the Campylobacter jejuni isolates carried the short operon (arsP, and arsR). The presence of arsC and acr3 did not significantly increase arsenic resistance with the exception of conferring resistance to higher concentrations of arsenate to some Campylobacter isolates. arsB was prevalent in 98% of the tested Campylobacter jejuni isolates, regardless of the presence or absence of arsC and acr3, but was completely absent in Campylobacter coli. To our knowledge, this is the first study to determine arsenic resistance and the prevalence of arsenic resistance genes in such a large number of Campylobacter isolates.

  15. Arsenic Resistance and Prevalence of Arsenic Resistance Genes in Campylobacter jejuni and Campylobacter coli Isolated from Retail Meats

    Directory of Open Access Journals (Sweden)

    Mohamed K. Fakhr

    2013-08-01

    Full Text Available Studies that investigate arsenic resistance in the foodborne bacterium Campylobacter are limited. A total of 552 Campylobacter isolates (281 Campylobacter jejuni and 271 Campylobacter coli isolated from retail meat samples were subjected to arsenic resistance profiling using the following arsenic compounds: arsanilic acid (4–2,048 μg/mL, roxarsone (4–2048 μg/mL, arsenate (16–8,192 μg/mL and arsenite (4–2,048 μg/mL. A total of 223 of these isolates (114 Campylobacter jejuni and 109 Campylobacter coli were further analyzed for the presence of five arsenic resistance genes (arsP, arsR, arsC, acr3, and arsB by PCR. Most of the 552 Campylobacter isolates were able to survive at higher concentrations of arsanilic acid (512–2,048 μg/mL, roxarsone (512–2,048 μg/mL, and arsenate (128–1,024 μg/mL, but at lower concentrations for arsenite (4–16 μg/mL. Ninety seven percent of the isolates tested by PCR showed the presence of arsP and arsR genes. While 95% of the Campylobacter coli isolates contained a larger arsenic resistance operon that has all of the four genes (arsP, arsR, arsC and acr3, 85% of the Campylobacter jejuni isolates carried the short operon (arsP, and arsR. The presence of arsC and acr3 did not significantly increase arsenic resistance with the exception of conferring resistance to higher concentrations of arsenate to some Campylobacter isolates. arsB was prevalent in 98% of the tested Campylobacter jejuni isolates, regardless of the presence or absence of arsC and acr3, but was completely absent in Campylobacter coli. To our knowledge, this is the first study to determine arsenic resistance and the prevalence of arsenic resistance genes in such a large number of Campylobacter isolates.

  16. Arsenic Resistance and Prevalence of Arsenic Resistance Genes in Campylobacter jejuni and Campylobacter coli Isolated from Retail Meats

    Science.gov (United States)

    Noormohamed, Aneesa; Fakhr, Mohamed K.

    2013-01-01

    Studies that investigate arsenic resistance in the foodborne bacterium Campylobacter are limited. A total of 552 Campylobacter isolates (281 Campylobacter jejuni and 271 Campylobacter coli) isolated from retail meat samples were subjected to arsenic resistance profiling using the following arsenic compounds: arsanilic acid (4–2,048 μg/mL), roxarsone (4–2048 μg/mL), arsenate (16–8,192 μg/mL) and arsenite (4–2,048 μg/mL). A total of 223 of these isolates (114 Campylobacter jejuni and 109 Campylobacter coli) were further analyzed for the presence of five arsenic resistance genes (arsP, arsR, arsC, acr3, and arsB) by PCR. Most of the 552 Campylobacter isolates were able to survive at higher concentrations of arsanilic acid (512–2,048 μg/mL), roxarsone (512–2,048 μg/mL), and arsenate (128–1,024 μg/mL), but at lower concentrations for arsenite (4–16 μg/mL). Ninety seven percent of the isolates tested by PCR showed the presence of arsP and arsR genes. While 95% of the Campylobacter coli isolates contained a larger arsenic resistance operon that has all of the four genes (arsP, arsR, arsC and acr3), 85% of the Campylobacter jejuni isolates carried the short operon (arsP, and arsR). The presence of arsC and acr3 did not significantly increase arsenic resistance with the exception of conferring resistance to higher concentrations of arsenate to some Campylobacter isolates. arsB was prevalent in 98% of the tested Campylobacter jejuni isolates, regardless of the presence or absence of arsC and acr3, but was completely absent in Campylobacter coli. To our knowledge, this is the first study to determine arsenic resistance and the prevalence of arsenic resistance genes in such a large number of Campylobacter isolates. PMID:23965921

  17. Identification of a protein which differs in lines of barley resistant or susceptible to Erysiphe graminis f. sp. hordei

    Energy Technology Data Exchange (ETDEWEB)

    Simons, S.P.; Somerville, S.C. (Michigan State Univ., East Lansing (USA))

    1990-05-01

    As yet no resistance genes or gene products have been isolated from a plant. We wish to isolate the Ml-a gene which encodes resistance by barley to Erysiphe graminis f. sp. hordei, the causal agent of the powdery mildew disease. We have utilized near isogenic barley lines, resistant and susceptible to E. g. hordei, race CR3 to provide a homogenous background for examination of protein differences related to resistance. To enrich for epidermal tissue, the site of infection, proteins from {sup 35}S labelled coleoptiles were analyzed by 2D-PAGE. Two major protein differences were observed, one of which is a polymorphism identified by a higher molecular weight form in the susceptible line, with the lower molecular weight form appearing in the resistant line. A partially susceptible mutant line derived from the resistant line shows a 63% reduction in the level of the resistant form of the polypeptide. A likely conclusion from these results is that we have identified the Ml-a gene product. Further characterization of the polymorphic protein and its gene will lead to an understanding of gene-for-gene related resistance processes.

  18. Multilocus Sequence Typing And Antibiotic Resistance Of Staphylococcus Aureus Isolated From The Brazilian Dairy Industry

    DEFF Research Database (Denmark)

    Dittmann, Karen Kiesbye; Chaul, Luiza; Lee, Sarah

    2015-01-01

    was the dominant CC in the investigated dairy plants. However, there were no indications of re-occurring (persistent) STs in the plants. The potential health risk of the isolates was assessed by antibiotic resistance and hemolytic activity screening. Resistance levels were low, and all of the isolates were...... presumptive methicillin-sensitive S. aureus. All of the isolates expressed hemolytic activity. The frequent isolation of CC1 strains in Brazilian dairy plants indicates, despite antibiotic sensitivity, a potential health risk to the human consumer....

  19. Quinolone and macrolide resistance in Campylobacter jejuni and C-coli: Resistance mechanisms and trends in human isolates

    DEFF Research Database (Denmark)

    Engberg, J.; Aarestrup, Frank Møller; Taylor, D. E.

    2001-01-01

    The incidence of human Campylobacter jejuni and C. coli infections has increased markedly in many parts of the world in the last decade as has the number of quinolone-resistant and, to a lesser extent, macrolide-resistant Campylobacter strains causing infections. We review macrolide and quinolone...... resistance in Campylobacter and track resistance trends in human clinical isolates in relation to use of these agents in food animals. Susceptibility data suggest that erythromycin and other macrolides should remain the drugs of choice in most regions, with systematic surveillance and central measures...... maintained, but fluoroquinolones may now be of limited use in the empiric treatment of Campylobacter infections in many regions....

  20. The behaviour of whey protein isolate in protecting Lactobacillus plantarum.

    Science.gov (United States)

    Khem, Sarim; Small, Darryl M; May, Bee K

    2016-01-01

    There is increasing evidence that whey protein isolates (WPI), can be utilised to encapsulate and protect bioactive substances, including lactic acid bacteria, due to their physicochemical properties. However, little is known about what happens in the immediate vicinity of the cells. This study examined the protective behaviour of WPI for two strains of Lactobacillus plantarum, A17 and B21, during spray drying. B21 was found to be more hydrophobic than A17 and required 50% of the amount of WPI to provide comparably high survival (∼ 90%). We hypothesise that WPI protects the hydrophobic bacteria by initial attachment to the unfolded whey protein due to hydrophobic interactions followed by adhesion to the proteins, resulting in cells being embedded within the walls of the capsules. The encapsulated strains had a moisture content of approximately 5.5% and during storage trials at 20 °C retained viability for at least eight weeks.

  1. [A rare mechanism of resistance to colistin in Escherichia coli isolated from raw poultry meat].

    Science.gov (United States)

    Karpíšková, Renáta; Koláčková, Ivana; Gelbíčová, Tereza; Zobaníková, Marie

    2017-06-01

    Plasmid-mediated resistance to colistin is a recently described phenomenon. The study reports this new type of colistin resistance in food isolates of Escherichia coli in the Czech Republic. Strains with phenotypically determined colistin resistance were studied for presence of the mcr-1 and mcr-2 genes. A positive finding of E. coli harboring the mcr-1 gene was confirmed in a sample of raw minced turkey meat imported from Poland. Two different strains of E. coli carrying the mcr-1 gene were detected in the same sample. This is the first reported case of this type of resistance in E. coli strains isolated from foods at retail in the Czech Republic.

  2. Direct repeat unit (dru) typing and antimicrobial resistance of methicillin-resistant Staphylococcus pseudintermedius isolated from dogs in Atlantic Canada.

    Science.gov (United States)

    Saab, Matthew E; Weese, J Scott; McClure, J T

    2017-07-01

    There are few reports investigating the characterization of methicillin-resistant Staphylococcus pseudintermedius (MRSP) in dogs in Canada and none from Atlantic Canada. The objectives of this study were to strain type MRSP isolates cultured at a regional diagnostic laboratory using direct repeat unit (dru) typing and to describe their antimicrobial resistance profiles. Ninety-four isolates recovered from dogs between 2010 and 2012 had dru typing, cluster analysis, and antimicrobial susceptibility testing done. The majority of isolates belonged to type dt11a (30.9%), dt10h (24.5%), dt9a (18.1%), and dt11af (10.6%) with the remaining 15.9% of isolates distributed among 13 dru types. The predominant dru types identified were similar in Ontario; however, cluster 9a appears to be less common in Atlantic Canada. A significant difference in the distribution of clusters among Atlantic provinces was detected (P = 0.01). Resistance to ≥ 2 non-β-lactam antimicrobials was observed in 71.4% of the isolates. The MRSP isolates from this study were notably less resistant than those reported in the literature. A more comprehensive study of the MRSP dru types could help further elucidate the distribution of this pathogen in Canada.

  3. Efflux pump gene expression in multidrug-resistant Mycobacterium tuberculosis clinical isolates.

    Science.gov (United States)

    Li, Guilian; Zhang, Jingrui; Guo, Qian; Jiang, Yi; Wei, Jianhao; Zhao, Li-li; Zhao, Xiuqin; Lu, Jianxin; Wan, Kanglin

    2015-01-01

    Isoniazid (INH) and rifampicin (RIF) are the two most effective drugs in tuberculosis therapy. Understanding the molecular mechanisms of resistance to these two drugs is essential to quickly diagnose multidrug-resistant (MDR) tuberculosis and extensive drug-resistant tuberculosis. Nine clinical Mycobacterium tuberculosis isolates resistant to only INH and RIF and 10 clinical pan-sensitive isolates were included to evaluate the expression of 20 putative drug efflux pump genes and sequence mutations in rpoB (RIF), katG (INH), the inhA promoter (INH), and oxyR-ahpC (INH). Nine and three MDR isolates were induced to overexpress efflux pump genes by INH and RIF, respectively. Eight and two efflux pump genes were induced to overexpress by INH and RIF in MDR isolates, respectively. drrA, drrB, efpA, jefA (Rv2459), mmr, Rv0849, Rv1634, and Rv1250 were overexpressed under INH or RIF stress. Most efflux pump genes were overexpressed under INH stress in a MDR isolates that carried the wild-type katG, inhA, and oxyR-ahpC associated with INH resistance than in those that carried mutations. The expression levels of 11 genes (efpA, Rv0849, Rv1250, P55 (Rv1410c), Rv1634, Rv2994, stp, Rv2459, pstB, drrA, and drrB) without drug inducement were significantly higher (P < 0.05) in nine MDR isolates than in 10 pan-sensitive isolates. In conclusion, efflux pumps may play an important role in INH acquired resistance in MDR M. tuberculosis, especially in those strains having no mutations in genes associated with INH resistance; basal expression levels of some efflux pump genes are higher in MDR isolates than in pan-sensitive isolates and the basal expressional differences may be helpful to diagnose and treat resistant tuberculosis.

  4. Efflux pump gene expression in multidrug-resistant Mycobacterium tuberculosis clinical isolates.

    Directory of Open Access Journals (Sweden)

    Guilian Li

    Full Text Available Isoniazid (INH and rifampicin (RIF are the two most effective drugs in tuberculosis therapy. Understanding the molecular mechanisms of resistance to these two drugs is essential to quickly diagnose multidrug-resistant (MDR tuberculosis and extensive drug-resistant tuberculosis. Nine clinical Mycobacterium tuberculosis isolates resistant to only INH and RIF and 10 clinical pan-sensitive isolates were included to evaluate the expression of 20 putative drug efflux pump genes and sequence mutations in rpoB (RIF, katG (INH, the inhA promoter (INH, and oxyR-ahpC (INH. Nine and three MDR isolates were induced to overexpress efflux pump genes by INH and RIF, respectively. Eight and two efflux pump genes were induced to overexpress by INH and RIF in MDR isolates, respectively. drrA, drrB, efpA, jefA (Rv2459, mmr, Rv0849, Rv1634, and Rv1250 were overexpressed under INH or RIF stress. Most efflux pump genes were overexpressed under INH stress in a MDR isolates that carried the wild-type katG, inhA, and oxyR-ahpC associated with INH resistance than in those that carried mutations. The expression levels of 11 genes (efpA, Rv0849, Rv1250, P55 (Rv1410c, Rv1634, Rv2994, stp, Rv2459, pstB, drrA, and drrB without drug inducement were significantly higher (P < 0.05 in nine MDR isolates than in 10 pan-sensitive isolates. In conclusion, efflux pumps may play an important role in INH acquired resistance in MDR M. tuberculosis, especially in those strains having no mutations in genes associated with INH resistance; basal expression levels of some efflux pump genes are higher in MDR isolates than in pan-sensitive isolates and the basal expressional differences may be helpful to diagnose and treat resistant tuberculosis.

  5. Role of embCAB gene mutations in ethambutol resistance in Mycobacterium tuberculosis isolates from India.

    Science.gov (United States)

    Jadaun, G P S; Das, Ram; Upadhyay, Prashant; Chauhan, D S; Sharma, V D; Katoch, V M

    2009-05-01

    In the present study, ethambutol (EMB) resistance-associated mutations were characterised in the embCAB genes of clinical isolates of Mycobacterium tuberculosis (MTB) collected in India. Thirty MTB isolates were tested for their susceptibility to first-line antitubercular drugs using the Löwenstein-Jensen proportion method, and EMB minimum inhibitory concentrations of MTB isolates were determined by the resazurin microtitre assay. Sequencing of various regions of the embCAB genes was performed to identify EMB resistance-associated mutations. Mutations of embB306 were detected in 15 of 23 EMB-resistant MTB isolates. Three EMB-resistant isolates had mutations at codon 270 of the embC gene, two of which also harboured embB306 mutations. No mutation was identified in the embA gene. All seven EMB-sensitive MTB isolates had the wild-type embCAB sequence. In summary, embB306 mutations were associated with EMB resistance, and mutation at codon 270 of the embC gene may contribute to high-level EMB resistance in some MTB isolates.

  6. Prevalence and Antimicrobial Resistance of Salmonella Isolated from Animal-Origin Food Items in Gondar, Ethiopia.

    Science.gov (United States)

    Ejo, Mebrat; Garedew, Legesse; Alebachew, Zabishwork; Worku, Walelgn

    2016-01-01

    Salmonella has been found to be the major cause of foodborne diseases and a serious public health problem in the world, with an increasing concern for the emergence and spread of antimicrobial-resistant strains. A cross-sectional study was conducted between February 2014 and December 2015 on food items of animal origin to assess the prevalence and antimicrobial resistance profiles of Salmonella isolates using standard bacteriological methods. The overall prevalence rate of 5.5% was recorded from the total analyzed food items of animal origin. Salmonella isolates were detected from 12% of raw meat, 8% of minced meat, 2.9% of burger samples, 18% of raw eggs, and 6% of raw milk. Furthermore, antimicrobial susceptibility test identified 47.6% resistant Salmonella isolates, 28.6% intermediately sensitive isolates, and 23.8% susceptible isolates. Among Salmonella isolates tested, 42.6%, 28.6%, and 14.3% were found to be relatively resistant to tetracycline, sulfamethoxazole-trimethoprim, and ampicillin, respectively, while 9.5%-19% were intermediately resistant to tetracycline, amoxicillin, ampicillin, cephalothin, and nitrofurantoin. Therefore, our findings provide the prevalence and drug resistance of Salmonella from foods of animal origin and contribute information to scientists as well as public health researchers to minimize the prevalent and resistant foodborne Salmonella species in Ethiopia.

  7. ANTIBIOTIC RESISTANCE IN LACTIC ACID BACTERIA ISOLATED FROM FERMENTED DAIRY PRODUCTS AND BOZA

    Directory of Open Access Journals (Sweden)

    Gamze Başbülbül

    2015-06-01

    Full Text Available In this study, the resistance of 83 strains of lactic acid bacteria isolated from Turkish cheese, yogurt, kefir and boza samples to 6 antibiotics (gentamicin, tetracycline, chloramphenicol, erythromycin, vancomycin and ciprofloxacin was evaluated. The 83 isolates were identified by 16S rRNA gene sequencing and according to BLAST comparisons with sequences in the data banks, those strains showing the highest similarities with the isolates were Enterococcus faecium (10, Lactococcus lactis subsp. Lactis (10, Lactobacillus fermentum (6, Lactobacillus plantarum (6, Lactobacillus coryniformis (7, Lactobacillus casei (13, Leuconostoc mesenteroides (14, Pediococcus pentosaceus (10, Weisella confusa (7. Antimicrobial resistance of strains to 6 antibiotics was determined using the agar dilution method. The antibiotic resistance among all the isolates was detected against chloramphenicol (31,3 % of the isolates, tetracycline (30,1 %, erythromycin (2,4 %, ciprofloxacin (2,41%, vancomycin (73,5 %, intrinsic resistance. Overall 19,3 % of the isolates showed resistance against multiple antibiotics. Antibiotic resistance genes were studied by PCR and the following genes were detected; tet(M gene in Lactobacillus fermentum (1, Lactobacillus plantarum (1, Pediococcus pentosaceus (5, Enterococcus faecium (2, Weisella confusa (4 and the vancomycin resistance gene van(A in one Weisella confusa strain.

  8. Prevalence and Antimicrobial Resistance of Salmonella Isolated from Animal-Origin Food Items in Gondar, Ethiopia

    Science.gov (United States)

    Garedew, Legesse; Alebachew, Zabishwork; Worku, Walelgn

    2016-01-01

    Salmonella has been found to be the major cause of foodborne diseases and a serious public health problem in the world, with an increasing concern for the emergence and spread of antimicrobial-resistant strains. A cross-sectional study was conducted between February 2014 and December 2015 on food items of animal origin to assess the prevalence and antimicrobial resistance profiles of Salmonella isolates using standard bacteriological methods. The overall prevalence rate of 5.5% was recorded from the total analyzed food items of animal origin. Salmonella isolates were detected from 12% of raw meat, 8% of minced meat, 2.9% of burger samples, 18% of raw eggs, and 6% of raw milk. Furthermore, antimicrobial susceptibility test identified 47.6% resistant Salmonella isolates, 28.6% intermediately sensitive isolates, and 23.8% susceptible isolates. Among Salmonella isolates tested, 42.6%, 28.6%, and 14.3% were found to be relatively resistant to tetracycline, sulfamethoxazole-trimethoprim, and ampicillin, respectively, while 9.5%–19% were intermediately resistant to tetracycline, amoxicillin, ampicillin, cephalothin, and nitrofurantoin. Therefore, our findings provide the prevalence and drug resistance of Salmonella from foods of animal origin and contribute information to scientists as well as public health researchers to minimize the prevalent and resistant foodborne Salmonella species in Ethiopia. PMID:28074185

  9. Drug resistance of Mycobacterium tuberculosis isolates from tuberculosis lymphadenitis patients in Ethiopia

    Directory of Open Access Journals (Sweden)

    Fantahun Biadglegne

    2014-01-01

    Full Text Available Background & objectives: The emergence of drug resistance tuberculosis (TB is a significant challenge for TB control and prevention programmes, and the major problem is multidrug resistant tuberculosis (MDR-TB. The present study was carried out to determine the frequency of drug resistant Mycobacterium tuberculosis isolates among newly and retreated TB lymphadenitis patients and risk factors for acquiring this infection. Methods: Two hundred twenty five M. tuberculosis isolates from TB lymphadenitis patients who were diagnosed as new and retreated tuberculosis cases between April 2012 and May 2012 were included in this study. Isolates were tested for susceptibility to isoniazed (INH, rifampicin (RMP, streptomycin (SM, ethambutol (EMB and pyrazinamide (PZA using the BacT/AlerT 3D system protocol. Results: Among 225 isolates, 15 (6.7% were resistant to at least one first line anti-TB drug. Three (1.3% were MDR-TB. Resistance to INH, RMP, SM, and EMB was found in 8 (3.6%, 4 (1.8%, 10 (4.4%, and 4 (1.8% isolates, respectively. Of the 212 new TB lymphadenitis cases three (1.4% were MDR-TB. A rifampicin resistant M. tuberculosis isolate was diagnosed from smear and culture negative newly treated cases. All isolates were susceptible to PZA. Matted cervical lymph nodes were the prominent sites involved. Newly treated TB lymphadenitis patients had a greater risk for presenting resistance to anti-TB drugs ( p =0.046. Interpretation & conclusions: Our study showed that TB lymphadenitis patients harboured drug resistant TB and MDR-TB, although at a low rate. Resistance was not associated with age, sex, patients′ education and contact history. Further research is required to determine transmission dynamics of drug resistant strains.

  10. Antibiotic Resistance Patterns of Escherichia coli Isolates from Hospitals in Kumasi, Ghana.

    Science.gov (United States)

    George, Duredoh Freeman; Gbedema, Stephen Yao; Agyare, Christian; Adu, Francis; Boamah, Vivian Etsiapa; Tawiah, Adelaide Ama; Saana, Sixtus Bieranye Bayaa Martin

    2012-01-01

    Nosocomial infections are infections acquired by a patient as a result of treatment in a hospital or healthcare service providing center and symptoms occurs within a short period of hospitalization. The study was to determine the antibiotic resistance patterns of Escherichia coli isolated from Kumasi-South, Tafo and Suntreso Hospitals, Kumasi, Ghana. Total of 600 swabs samples from the hospitals were collected between January and June, 2010. The isolates were identified using morphological and biochemical means. A total of 97 E. coli isolates were obtained from the hospitals. Beds in hospital wards had the highest number of E. coli strains (53.6%), followed by floors (20.6%) while drainages had the least isolates (3.1%). Majority of the E. coli isolates (90.7%) exhibited resistance to ampicillin while 6.2 and 3.1% showed intermediate and sensitive respectively. Co-trimoxazole, 78.4% of the isolates were resistant while 9.3 and 12.4% exhibited intermediate and sensitive responses respectively. E. coli isolates (28.6 to 46.4%) were resistant to gentamicin, ciprofloxacin and ceftriaxone while 14.4 to 47.4% gave intermediate responses. Most isolates (80.4%) exhibited multi-drug resistance. There is a need to observe proper personal hygiene, use of effective disinfectants and proper disposal of contaminated/pathogenic materials in these hospitals to control nosocomial infections.

  11. Multidrug-resistant Salmonellae isolated in Japanese quails reared in Abeokuta, Nigeria.

    Science.gov (United States)

    Omoshaba, Ezekiel O; Olufemi, F O; Ojo, O E; Sonibare, A O; Agbaje, M

    2017-07-17

    Salmonellosis is a major bacterial disease causing huge economic losses in the poultry industry worldwide. This study was carried out to determine the period prevalence and antimicrobial susceptibility of Salmonella enterica in Japanese quails in Abeokuta, Nigeria. Four hundred cloacal swabs of quail birds were collected from 4 locations within Abeokuta. Salmonella was isolated from the samples using conventional methods for selective isolation of Salmonella and biochemical identification. Isolates were confirmed by polymerase chain reaction assays for the amplification and detection of Salmonella-associated virulence genes (invA and stn) using specific primers. Antimicrobial susceptibility testing was done using the Kirby-Bauer disk diffusion method. In all, Salmonella was isolated from 14 (3.5%) cloacal swabs. All 14 isolates possessed invA and stn genes. The Salmonella isolates showed resistance to tetracycline (100%), doxycycline (100%), ampicillin (100%), sulphamethoxazole (92.9%), nalidixic acid (85.8%), ceftazidime (78.6%), neomycin (64.3%), streptomycin (50%) and gentamycin (28.6%) but all the isolates were susceptible to ciprofloxacin. The isolates were resistant to at least three antimicrobials indicating multidrug resistance. The results concluded that Japanese quails harbour multidrug-resistant Salmonella which could be transmitted to humans through consumption of contaminated food or by direct and indirect contact with the carrier birds. Antimicrobial resistance could be due to overdependence on antimicrobials. Ciprofloxacin could be considered in the treatment of zoonotic Salmonellosis in humans.

  12. Investigation of antimicrobial resistance in Escherichia coli and enterococci isolated from Tibetan pigs.

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    Peng Li

    Full Text Available OBJECTIVES: This study investigated the antimicrobial resistance of Escherichia coli and enterococci isolated from free-ranging Tibetan pigs in Tibet, China, and analyzed the influence of free-ranging husbandry on antimicrobial resistance. METHODS: A total of 232 fecal samples were collected from Tibetan pigs, and the disk diffusion method was used to examine their antimicrobial resistance. Broth microdilution and agar dilution methods were used to determine minimum inhibitory concentrations for antimicrobial agents for which disks were not commercially available. RESULTS: A total of 129 E. coli isolates and 84 Enterococcus isolates were recovered from the fecal samples. All E. coli isolates were susceptible to amoxicillin/clavulanic acid, and 40.4% were resistant to tetracycline. A small number of isolates were resistant to florfenicol (27.9%, ampicillin (27.9%, sulfamethoxazole/trimethoprim (19.4%, nalidixic acid (19.4%, streptomycin (16.2% and ceftiofur (10.9%, and very low resistance rates to ciprofloxacin (7.8%, gentamicin (6.9%, and spectinomycin (2.3% were observed in E. coli. All Enterococcus isolates, including E. faecium, E. faecalis, E. hirae, and E. mundtii, were susceptible to amoxicillin/clavulanic acid and vancomycin, but showed high frequencies of resistance to oxacillin (92.8%, clindamycin (82.1%, tetracycline (64.3%, and erythromycin (48.8%. Resistance rates to florfenicol (17.9%, penicillin (6.0%, ciprofloxacin (3.6%, levofloxacin (1.2%, and ampicillin (1.2% were low. Only one high-level streptomycin resistant E. faecium isolate and one high-level gentamicin resistant E. faecium isolate were observed. Approximately 20% and 70% of E. coli and Enterococcus isolates, respectively, were defined as multidrug-resistant. CONCLUSIONS: In this study, E. coli and Enterococcus isolated from free-ranging Tibetan pigs showed relatively lower resistance rates than those in other areas of China, where more intensive farming practices are

  13. Investigation of antimicrobial resistance in Escherichia coli and enterococci isolated from Tibetan pigs.

    Science.gov (United States)

    Li, Peng; Wu, Dongfang; Liu, Kunyao; Suolang, Sizhu; He, Tao; Liu, Xuan; Wu, Congming; Wang, Yang; Lin, Degui

    2014-01-01

    This study investigated the antimicrobial resistance of Escherichia coli and enterococci isolated from free-ranging Tibetan pigs in Tibet, China, and analyzed the influence of free-ranging husbandry on antimicrobial resistance. A total of 232 fecal samples were collected from Tibetan pigs, and the disk diffusion method was used to examine their antimicrobial resistance. Broth microdilution and agar dilution methods were used to determine minimum inhibitory concentrations for antimicrobial agents for which disks were not commercially available. A total of 129 E. coli isolates and 84 Enterococcus isolates were recovered from the fecal samples. All E. coli isolates were susceptible to amoxicillin/clavulanic acid, and 40.4% were resistant to tetracycline. A small number of isolates were resistant to florfenicol (27.9%), ampicillin (27.9%), sulfamethoxazole/trimethoprim (19.4%), nalidixic acid (19.4%), streptomycin (16.2%) and ceftiofur (10.9%), and very low resistance rates to ciprofloxacin (7.8%), gentamicin (6.9%), and spectinomycin (2.3%) were observed in E. coli. All Enterococcus isolates, including E. faecium, E. faecalis, E. hirae, and E. mundtii, were susceptible to amoxicillin/clavulanic acid and vancomycin, but showed high frequencies of resistance to oxacillin (92.8%), clindamycin (82.1%), tetracycline (64.3%), and erythromycin (48.8%). Resistance rates to florfenicol (17.9%), penicillin (6.0%), ciprofloxacin (3.6%), levofloxacin (1.2%), and ampicillin (1.2%) were low. Only one high-level streptomycin resistant E. faecium isolate and one high-level gentamicin resistant E. faecium isolate were observed. Approximately 20% and 70% of E. coli and Enterococcus isolates, respectively, were defined as multidrug-resistant. In this study, E. coli and Enterococcus isolated from free-ranging Tibetan pigs showed relatively lower resistance rates than those in other areas of China, where more intensive farming practices are used. These results also revealed that free

  14. Multiple drug resistance protein (MDR-1, multidrug resistance-related protein (MRP and lung resistance protein (LRP gene expression in childhood acute lymphoblastic leukemia

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    Elvis Terci Valera

    Full Text Available CONTEXT: Despite the advances in the cure rate for acute lymphoblastic leukemia, approximately 25% of affected children suffer relapses. Expression of genes for the multiple drug resistance protein (MDR-1, multidrug resistance-related protein (MRP, and lung resistance protein (LRP may confer the phenotype of resistance to the treatment of neoplasias. OBJECTIVE: To analyze the expression of the MDR-1, MRP and LRP genes in children with a diagnosis of acute lymphoblastic leukemia via the semiquantitative reverse transcription polymerase chain reaction (RT-PCR, and to determine the correlation between expression and event-free survival and clinical and laboratory variables. DESIGN: A retrospective clinical study. SETTING: Laboratory of Pediatric Oncology, Department of Pediatrics, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Brazil. METHODS: Bone marrow aspirates from 30 children with a diagnosis of acute lymphoblastic leukemia were assessed for the expression of messenger RNA for the MDR-1, MRP and LRP genes by semi-quantitative RT-PCR. RESULTS: In the three groups studied, only the increased expression of LRP was related to worsened event-free survival (p = 0.005. The presence of the common acute lymphoblastic leukemia antigen (CALLA was correlated with increased LRP expression (p = 0.009 and increased risk of relapse or death (p = 0.05. The relative risk of relapse or death was six times higher among children with high LRP expression upon diagnosis (p = 0.05, as confirmed by multivariate analysis of the three genes studied (p = 0.035. DISCUSSION: Cell resistance to drugs is a determinant of the response to chemotherapy and its detection via RT-PCR may be of clinical importance. CONCLUSIONS: Evaluation of the expression of genes for resistance to antineoplastic drugs in childhood acute lymphoblastic leukemia upon diagnosis, and particularly the expression of the LRP gene, may be of clinical relevance, and should be the

  15. Diversity and antibiotic resistance patterns of Sphingomonadaceae isolates from drinking water.

    Science.gov (United States)

    Vaz-Moreira, Ivone; Nunes, Olga C; Manaia, Célia M

    2011-08-15

    Sphingomonadaceae (n = 86) were isolated from a drinking water treatment plant (n = 6), tap water (n = 55), cup fillers for dental chairs (n = 21), and a water demineralization filter (n = 4). The bacterial isolates were identified based on analysis of the 16S rRNA gene sequence, and intraspecies variation was assessed on the basis of atpD gene sequence analysis. The isolates were identified as members of the genera Sphingomonas (n = 27), Sphingobium (n = 28), Novosphingobium (n = 12), Sphingopyxis (n = 7), and Blastomonas (n = 12). The patterns of susceptibility to five classes of antibiotics were analyzed and compared for the different sites of isolation and taxonomic groups. Colistin resistance was observed to be intrinsic (92%). The highest antibiotic resistance prevalence values were observed in members of the genera Sphingomonas and Sphingobium and for beta-lactams, ciprofloxacin, and cotrimoxazole. In tap water and in water from dental chairs, antibiotic resistance was more prevalent than in the other samples, mainly due to the predominance of isolates of the genera Sphingomonas and Sphingobium. These two genera presented distinct patterns of association with antibiotic resistance, suggesting different paths of resistance development. Antibiotic resistance patterns were often related to the species rather than to the site or strain, suggesting the importance of vertical resistance transmission in these bacteria. This is the first study demonstrating that members of the family Sphingomonadaceae are potential reservoirs of antibiotic resistance in drinking water.

  16. Multidrug-Resistant Salmonella Isolates from Swine in the Eastern Cape Province, South Africa.

    Science.gov (United States)

    Iwu, Chinwe Juliana; Iweriebor, Benson Chuks; Obi, Larry Chikwelu; Basson, Albertus Kotze; Okoh, Anthony Ifeanyi

    2016-07-01

    The exposure of farm animals to antimicrobials for treatment, prophylaxis, or growth promotion can select for resistant bacteria that can be transmitted to humans, and Salmonella as an important zoonotic pathogen can act as a potential reservoir of antimicrobial resistance determinants. We assessed the antibiogram profiles of Salmonella species isolated from pig herds in two commercial farms in South Africa. Two hundred fifty-eight presumptive Salmonella isolates were recovered from the fecal samples of 500 adult pigs. Specific primers targeting Salmonella serogroups A, B, C1, C2, and D were used to determine the prevalence of different serogroups. Only serogroup A (n = 48) was detected, while others were not. Antimicrobial susceptibility of the confirmed Salmonella serogroup A isolates was performed by using the disk diffusion method against a panel of 18 antibiotics. All the 48 isolates were resistant to tetracycline and oxytetracycline, while 75% were resistant to ampicillin, sulphamethoxazole-trimethoprim, nalidixic acid, and streptomycin. All the isolates exhibited multidrug resistance, with the predominant phenotype being against 11 antibiotics, and multiple antibiotic resistance index ranged between 0.3 and 0.6. The incidence of genes encoding resistance against ampicillin (ampC), tetracycline (tetA), and streptomycin (strA) were 54, 61, and 44%, respectively. We conclude that healthy pigs are potential reservoirs of multidrug-resistant Salmonella that could be transmitted to humans through the food chain and, hence, a significant public health threat.

  17. Diversity and Antibiotic Resistance Patterns of Sphingomonadaceae Isolates from Drinking Water▿

    Science.gov (United States)

    Vaz-Moreira, Ivone; Nunes, Olga C.; Manaia, Célia M.

    2011-01-01

    Sphingomonadaceae (n = 86) were isolated from a drinking water treatment plant (n = 6), tap water (n = 55), cup fillers for dental chairs (n = 21), and a water demineralization filter (n = 4). The bacterial isolates were identified based on analysis of the 16S rRNA gene sequence, and intraspecies variation was assessed on the basis of atpD gene sequence analysis. The isolates were identified as members of the genera Sphingomonas (n = 27), Sphingobium (n = 28), Novosphingobium (n = 12), Sphingopyxis (n = 7), and Blastomonas (n = 12). The patterns of susceptibility to five classes of antibiotics were analyzed and compared for the different sites of isolation and taxonomic groups. Colistin resistance was observed to be intrinsic (92%). The highest antibiotic resistance prevalence values were observed in members of the genera Sphingomonas and Sphingobium and for beta-lactams, ciprofloxacin, and cotrimoxazole. In tap water and in water from dental chairs, antibiotic resistance was more prevalent than in the other samples, mainly due to the predominance of isolates of the genera Sphingomonas and Sphingobium. These two genera presented distinct patterns of association with antibiotic resistance, suggesting different paths of resistance development. Antibiotic resistance patterns were often related to the species rather than to the site or strain, suggesting the importance of vertical resistance transmission in these bacteria. This is the first study demonstrating that members of the family Sphingomonadaceae are potential reservoirs of antibiotic resistance in drinking water. PMID:21705522

  18. Molecular mechanisms of fluconazole resistance in Candida parapsilosis isolates from a U.S. surveillance system.

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    Grossman, Nina T; Pham, Cau D; Cleveland, Angela A; Lockhart, Shawn R

    2015-02-01

    Candida parapsilosis is the second or third most common cause of candidemia in many countries. The Infectious Diseases Society of America recommends fluconazole as the primary therapy for C. parapsilosis candidemia. Although the rate of fluconazole resistance among C. parapsilosis isolates is low in most U.S. institutions, the resistance rate can be as high as 7.5%. This study was designed to assess the mechanisms of fluconazole resistance in 706 incident bloodstream isolates from U.S. hospitals. We sequenced the ERG11 and MRR1 genes of 122 C. parapsilosis isolates with resistant (30 isolates; 4.2%), susceptible dose-dependent (37 isolates; 5.2%), and susceptible (55 isolates) fluconazole MIC values and used real-time PCR of RNA from 17 isolates to investigate the regulation of MDR1. By comparing these isolates to fully fluconazole-susceptible isolates, we detected at least two mechanisms of fluconazole resistance: an amino acid substitution in the 14-α-demethylase gene ERG11 and overexpression of the efflux pump MDR1, possibly due to point mutations in the MRR1 transcription factor that regulates MDR1. The ERG11 single nucleotide polymorphism (SNP) was found in 57% of the fluconazole-resistant isolates and in no susceptible isolates. The MRR1 SNPs were more difficult to characterize, as not all resulted in overexpression of MDR1 and not all MDR1 overexpression was associated with an SNP in MRR1. Further work to characterize the MRR1 SNPs and search for overexpression of other efflux pumps is needed.

  19. Exploring the contribution of efflux on the resistance to fluoroquinolones in clinical isolates of Staphylococcus aureus

    Science.gov (United States)

    2011-01-01

    Background Antimicrobial resistance mediated by efflux systems is still poorly characterized in Staphylococcus aureus, despite the description of several efflux pumps (EPs) for this bacterium. In this work we used several methodologies to characterize the efflux activity of 52 S. aureus isolates resistant to ciprofloxacin collected in a hospital in Lisbon, Portugal, in order to understand the role played by these systems in the resistance to fluoroquinolones. Results Augmented efflux activity was detected in 12 out of 52 isolates and correlated with increased resistance to fluoroquinolones. Addition of efflux inhibitors did not result in the full reversion of the fluoroquinolone resistance phenotype, yet it implied a significant decrease in the resistance levels, regardless of the type(s) of mutation(s) found in the quinolone-resistance determining region of grlA and gyrA genes, which accounted for the remaining resistance that was not efflux-mediated. Expression analysis of the genes coding for the main efflux pumps revealed increased expression only in the presence of inducing agents. Moreover, it showed that not only different substrates can trigger expression of different EP genes, but also that the same substrate can promote a variable response, according to its concentration. We also found isolates belonging to the same clonal type that showed different responses towards drug exposure, thus evidencing that highly related clinical isolates may diverge in the efflux-mediated response to noxious agents. The data gathered by real-time fluorometric and RT-qPCR assays suggest that S. aureus clinical isolates may be primed to efflux antimicrobial compounds. Conclusions The results obtained in this work do not exclude the importance of mutations in resistance to fluoroquinolones in S. aureus, yet they underline the contribution of efflux systems for the emergence of high-level resistance. All together, the results presented in this study show the potential role

  20. Inducible clindamycin resistance in Staphylococcus aureus isolates recovered from Mashhad, Iran

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    Najmeh seifi

    2012-06-01

    Full Text Available Background and Objectives: Staphylococcus aureus is an important agent in hospital and community-associated infections, causing high morbidity and mortality. Introduction of the new antimicrobial classes for this pathogen is usually followed by the emergence of resistant strains through multiple mechanisms. For instance, resistance to clindamycin (CLI, can be constitutive or inducible. Inducible clindamycin resistance which may lead to treatment failure can simply be identified by performing D-test. The aim of this study was to determine the prevalence of inducible clindamycin resistance among Staphylococcus aureus isolates by D-test method.Materials and Methods: This was a cross-sectional study conducted on 254 non-duplicated S. aureus isolates in Imam Reza hospital of Mashhad during 2010. Susceptibility to oxacillin, cefoxitin, erythromycin and clindamycin was performed by disk agar diffusion method according to CLSI guidelines and D-shaped clindamycin susceptibility patterns where considered as D-test positive (D+.Results: Of 211 S. aureus isolates 88 (37.82% were methicillin resistant. It was found that of 88 MRSA isolates, 78 (88.63% were erythromycin (ERY resistant and 46 (52.27% were CLI resistant. ERY and CLI resistance in MSSA strains was 21.95% and 11.96% respectively. Inducible clindamycin resistance was detected in 18 (20.45% MRSA isolates. 47(53.40% of MRSA isolates and 9 (7.32% of MSSA showed constitutive MLSB phenotype.Conclusion: In conclusion, we found a high prevalence of inducible clindamycin resistance phenotype in our region. We recommend that whenever clindamycin is intended for S. aureus infections, D-test should be performed to facilitate the optimal treatment of patients.Keywords: Staphylococcus aureus, clindamycin, Inducible resistance

  1. Exploring the contribution of efflux on the resistance to fluoroquinolones in clinical isolates of Staphylococcus aureus

    LENUS (Irish Health Repository)

    Costa, Sofia SANTOS

    2011-10-27

    Abstract Background Antimicrobial resistance mediated by efflux systems is still poorly characterized in Staphylococcus aureus, despite the description of several efflux pumps (EPs) for this bacterium. In this work we used several methodologies to characterize the efflux activity of 52 S. aureus isolates resistant to ciprofloxacin collected in a hospital in Lisbon, Portugal, in order to understand the role played by these systems in the resistance to fluoroquinolones. Results Augmented efflux activity was detected in 12 out of 52 isolates and correlated with increased resistance to fluoroquinolones. Addition of efflux inhibitors did not result in the full reversion of the fluoroquinolone resistance phenotype, yet it implied a significant decrease in the resistance levels, regardless of the type(s) of mutation(s) found in the quinolone-resistance determining region of grlA and gyrA genes, which accounted for the remaining resistance that was not efflux-mediated. Expression analysis of the genes coding for the main efflux pumps revealed increased expression only in the presence of inducing agents. Moreover, it showed that not only different substrates can trigger expression of different EP genes, but also that the same substrate can promote a variable response, according to its concentration. We also found isolates belonging to the same clonal type that showed different responses towards drug exposure, thus evidencing that highly related clinical isolates may diverge in the efflux-mediated response to noxious agents. The data gathered by real-time fluorometric and RT-qPCR assays suggest that S. aureus clinical isolates may be primed to efflux antimicrobial compounds. Conclusions The results obtained in this work do not exclude the importance of mutations in resistance to fluoroquinolones in S. aureus, yet they underline the contribution of efflux systems for the emergence of high-level resistance. All together, the results presented in this study show the potential

  2. Exploring the contribution of efflux on the resistance to fluoroquinolones in clinical isolates of Staphylococcus aureus

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    Costa Sofia

    2011-10-01

    Full Text Available Abstract Background Antimicrobial resistance mediated by efflux systems is still poorly characterized in Staphylococcus aureus, despite the description of several efflux pumps (EPs for this bacterium. In this work we used several methodologies to characterize the efflux activity of 52 S. aureus isolates resistant to ciprofloxacin collected in a hospital in Lisbon, Portugal, in order to understand the role played by these systems in the resistance to fluoroquinolones. Results Augmented efflux activity was detected in 12 out of 52 isolates and correlated with increased resistance to fluoroquinolones. Addition of efflux inhibitors did not result in the full reversion of the fluoroquinolone resistance phenotype, yet it implied a significant decrease in the resistance levels, regardless of the type(s of mutation(s found in the quinolone-resistance determining region of grlA and gyrA genes, which accounted for the remaining resistance that was not efflux-mediated. Expression analysis of the genes coding for the main efflux pumps revealed increased expression only in the presence of inducing agents. Moreover, it showed that not only different substrates can trigger expression of different EP genes, but also that the same substrate can promote a variable response, according to its concentration. We also found isolates belonging to the same clonal type that showed different responses towards drug exposure, thus evidencing that highly related clinical isolates may diverge in the efflux-mediated response to noxious agents. The data gathered by real-time fluorometric and RT-qPCR assays suggest that S. aureus clinical isolates may be primed to efflux antimicrobial compounds. Conclusions The results obtained in this work do not exclude the importance of mutations in resistance to fluoroquinolones in S. aureus, yet they underline the contribution of efflux systems for the emergence of high-level resistance. All together, the results presented in this study

  3. Evaluation of antibiotic resistance among isolated pathogenic bacteria from shrimp hatcheries in Bushehr province

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    Azam Moghimi

    2014-01-01

    Full Text Available Abstract Background: Rapid development of shrimp aquaculture has resulted in widespread use of antibiotics for preventing and curing diseases. In aquaculture, particularly shrimp hatcheries antibiotics are routinely used at therapeutic levels to treat disease and at sub-therapeutic levels as prophylactic agents to increase feed efficiency. Antibiotic residues in the environment are likely to lead to the development and maintenance of antibiotic resistance in microbial populations. The aim of this study was determine of antibiotic resistance to two shrimp pathogens Vibrio harveyi, V.alginolyticus, that they are agents of mortality in shrimp hatcheries. Material and Methods: After isolation and detection(by biochemical tests of two species of bacterial pathogens from three hatcheries of Bushehr province, bacterial strains were tested for sensitivity to antibiotics including erythromycin, streptomycin, oxytetracyclin, and trimetoprim by disk diffusion method. Results: Results showed that all isolated bacteria Vibrio harveyi from three hatcheries were sensitive to oxytetracyclin and trimetoprim, but to streptomycin were resistant, and to erythromycin in hatcheries A, B, C was intermediate, resistance, sensitive respectively. Bacteria Vibrio alginolyticus isolated from three hatcheries were resistant to streptomycin. But they isolated from a hatchery to the other antibiotics erythromycin, oxytetracyclin and trimetoprim were resistant, intermediate and intermediate, respectively. Also they isolated from B hatchery were resistant, sensitive and sensitive to erythromycin, oxytetracyclin and trimetoprim, respectively And from C hatchery were intermediate, sensitive and sensitive to antibiotics, respectively. Conclusion: Isolated bacteria showed the most resistance to streptomycin and erythromycin respectively. These antibiotics is used frequently in medicine and veterinary, with entrance of human and animal's bacteria resistance via waste and fluid water

  4. Clonal spread of antimicrobial-resistant Escherichia coli isolates among pups in two kennels

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    Takahashi Toshio

    2011-02-01

    Full Text Available Abstract Although the dog breeding industry is common in many countries, the presence of antimicrobial resistant bacteria among pups in kennels has been infrequently investigated. This study was conducted to better understand the epidemiology of antimicrobial-resistant Escherichia coli isolates from kennel pups not treated with antimicrobials. We investigated susceptibilities to 11 antimicrobials, and prevalence of extended-spectrum β-lactamase (ESBL in 86 faecal E. coli isolates from 43 pups in two kennels. Genetic relatedness among all isolates was assessed using pulsed-field gel electrophoresis (PFGE. Susceptibility tests revealed that 76% of the isolates were resistant to one or more of tested antimicrobials, with resistance to dihydrostreptomycin most frequently encountered (66.3% followed by ampicillin (60.5%, trimethoprim-sulfamethoxazole (41.9%, oxytetracycline (26.7%, and chloramphenicol (26.7%. Multidrug resistance, defined as resistance against two or more classes of antimicrobials, was observed in 52 (60.5% isolates. Three pups in one kennel harboured SHV-12 ESBL-producing isolates. A comparison between the two kennels showed that frequencies of resistance against seven antimicrobials and the variation in resistant phenotypes differed significantly. Analysis by PFGE revealed that clone sharing rates among pups of the same litters were not significantly different in both kennels (64.0% vs. 88.9%, whereas the rates among pups from different litters were significantly different between the two kennels (72.0% vs. 33.3%, P E. coli clones, including multidrug-resistant and ESBL-producing clones. It is likely that resistant and susceptible bacteria can clonally spread among the same and/or different litters thus affecting the resistance prevalence.

  5. Molecular detection of Rifampicin and Isoniazid resistance in culture isolates of newly diagnosed TB patients

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    Vanisree R, Kavitha Latha M, Neelima A, Prasanti

    2014-04-01

    Full Text Available Introduction: Multidrug-resistant tuberculosis (MDR-TB isan emerging public health problem in many regions of the world, particularly in developing nations. Accurate and rapid diagnosisis essential in the management of MDR-TB, not onlyto optimize treatment but also to prevent transmission. Aims: To evaluate drug resistance in culture isolates by conventional and molecular methods and detect drug resistance gene in MDR-TB patients. Material and Method: 100 newly diagnosed pulmonary tuberculosis (TB diagnosed patients attending TB Clinic, Gandhi Hospital, Secunderabad were included in the study. Two sputum samples collected from the patients were subjected to sputum microscopy, culture, Drug Susceptibility Testing (DST. Geno Type Mycobacterium Tuberculosis Drug Resistance (MTBDR plus assay was done on the culture isolates to detect Rifampicin and Isoniazid (INH resistance. Results: Out of 100 samples, 48 % smear positivity by Ziehl Neelsen (ZN method, 51 % culture positivity on LJ medium,11.7% multi drug resistance for Rifampicin and Isoniazid with conventional drug susceptibility – Proportion method,17.6 % drug resistance by molecular method – Geno Type MTBDR plus was observed. Among the 4 Rifampicin (Rif resistant isolates 2isolates showed mutation (mut at D516V and in other 2 isolates only wild type (WT was missing but no mut was seen . In the 1 Isoniazid (INH resistant isolate WT was missing, but no mutation was seen. Among the 4 Rif +INH resistance all showed mut at S531L for RIF and at S315T1. Conclusion: The Genotype MTBDR assay is a rapid and reliable tool for the routine direct detection of MTB strains and of strains resistant to INH and RIF in smear positive, highly infectious patients. The rapid turn around time of the test enables the optimization of the therapy of these patients before confirmatory culture results are available. The test does not require viable organisms and thus reduces the biohazard risk in the laboratory.

  6. Antibiotic susceptibility and molecular mechanisms of macrolide resistance in streptococci isolated from adult cystic fibrosis patients.

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    Thornton, Christina S; Grinwis, Margot E; Sibley, Christopher D; Parkins, Michael D; Rabin, Harvey R; Surette, Michael G

    2015-11-01

    The cystic fibrosis (CF) airways are colonized by polymicrobial communities with high bacterial load and are influenced by frequent antibiotic exposures. This community includes diverse streptococci, some of which have been directly or indirectly associated with pulmonary exacerbations. As many streptococci are naturally competent, horizontal transfer of antibiotic-resistant determinants coupled with frequent and/or chronic antibiotic exposure may contribute to high resistance rates. In this study, we assessed antibiotic resistance in 413 streptococcal isolates from adult CF patients against nine antibiotics relevant in CF treatment. We observed very low rates of cephalosporin resistance [cefepime and ceftriaxone ( resistance to tetracycline (∼34%) and sulfamethoxazole/trimethoprim (∼45%). The highest rate of antibiotic resistance was to the macrolides [azithromycin (56.4%) and erythromycin (51.6%)]. We also investigated the molecular mechanisms of macrolide resistance and found that only half of our macrolide-resistant streptococci isolates contained the mef (efflux pump) or erm (methylation of 23S ribosomal target site) genes. The majority of isolates were, however, found to have point mutations at position 2058 or 2059 of the 23S ribosomal subunit - a molecular mechanism of resistance not commonly reported in the non-pyogenic and non-pneumococcal streptococci, and unique in comparison with previous studies. The high rates of resistance observed here may result in poor outcomes where specific streptococci are contributing to CF airway disease and serve as a reservoir of resistance genes within the CF airway microbiome.

  7. Antibiotic resistant pattern of methicillin resistant and sensitive Staphylococcus aureus isolated from patients durining 2009-2010, Ahvaz, Iran.

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    N Parhizgari

    2013-12-01

    Full Text Available Abstract Background & aim: Staphylococcus aureus is one of the most important nosocomial infecting agents resistant to commonly used antibiotics. Nowadays, methicillin-resistant S. aureus (MRSA is considered one of the main causes of nosocomial infections. The aim of this study was to identify the antibiotic resistance pattern of methicicllin- resistant and susceptible strains in Ahwaz, Iran. Methods: In the present cross - sectional study, a number of 255 clinically suspected cases of Staphylococcus aureus were collected during a 19 month period. The bacteria were investigated using standard biochemical tests such as catalase, mannitol fermentation, coagulase and Dnase. Sensitive strains were confirmed by disk diffusion method compared to commonly used antibiotics. The collected data were analyzed using descriptive statistical tests. Results: of 255 suspected cases, 180 were confirmed as S.aureus, a total of 59 strains of S. aureus (2/37 percent were resistant to methicillin. Resistance to S. aureus strains resistant to methicillin included: chloramphenicol (3.38%, rifampin (45.76%, norfloxacin (89.83%, gentamicin (89.83%, ciprofloxacin, (91.52%, azithromycin, (88.13%, cotrimoxazole (86.44% and all isolates strains were sensitive to vancomycin and nitrofurantoin. A total of 10 different patterns of antibiotic resistance in methicillin-resistant Staphylococcus aureus strains were identified. Conclusion: Expression of new resistance factor in nosocomial infection is one of the major challenges in treating these infections. This study showed a high prevalence of resistance against some class of antibiotics in MRSA isolated from Imam Khomeini and Golestan hospital of Ahwaz, Iran. Key words: Nosocomial infection, Methicillin Resistant Staphylococcus aureus (MRSA, Antibiotic Resistant Pattern

  8. Resistance trends among clinical isolates in China reported from CHINET surveillance of bacterial resistance, 2005-2014.

    Science.gov (United States)

    Hu, F-P; Guo, Y; Zhu, D-M; Wang, F; Jiang, X-F; Xu, Y-C; Zhang, X-J; Zhang, C-X; Ji, P; Xie, Y; Kang, M; Wang, C-Q; Wang, A-M; Xu, Y-H; Shen, J-L; Sun, Z-Y; Chen, Z-J; Ni, Y-X; Sun, J-Y; Chu, Y-Z; Tian, S-F; Hu, Z-D; Li, J; Yu, Y-S; Lin, J; Shan, B; Du, Y; Han, Y; Guo, S; Wei, L-H; Wu, L; Zhang, H; Kong, J; Hu, Y-J; Ai, X-M; Zhuo, C; Su, D-H; Yang, Q; Jia, B; Huang, W

    2016-03-01

    With the aim of gathering temporal trends on bacterial epidemiology and resistance from multiple laboratories in China, the CHINET surveillance system was organized in 2005. Antimicrobial susceptibility testing was carried out according to a unified protocol using the Kirby-Bauer method or automated systems. Results were analyzed according to Clinical and Laboratory Standards Institute (CLSI) 2014 definitions. Between 2005 and 2014, the number of bacterial isolates ranged between 22,774 and 84,572 annually. Rates of extended-spectrum β-lactamase production among Escherichia coli isolates were stable, between 51.7 and 55.8%. Resistance of E. coli and Klebsiella pneumoniae to amikacin, ciprofloxacin, piperacillin/tazobactam and cefoperazone/sulbactam decreased with time. Carbapenem resistance among K. pneumoniae isolates increased from 2.4 to 13.4%. Resistance of Pseudomonas aeruginosa strains against all of antimicrobial agents tested including imipenem and meropenem decreased with time. On the contrary, resistance of Acinetobacter baumannii strains to carbapenems increased from 31 to 66.7%. A marked decrease of methicillin resistance from 69% in 2005 to 44.6% in 2014 was observed for Staphylococcus aureus. Carbapenem resistance rates in K. pneumoniae and A. baumannii in China are high. Our results indicate the importance of bacterial surveillance studies.

  9. Antimicrobial resistance and virulence profiles of Salmonella isolated from butcher shops in Minas Gerais, Brazil.

    Science.gov (United States)

    Cossi, Marcus Vinícius Coutinho; Burin, Raquel Cristina Konrad; Lopes, Danilo Augusto; Dias, Mariane Rezende; Castilho, Natalia Parma Augusto de; de Arruda Pinto, Paulo Sérgiode; Nero, Luís Augusto

    2013-09-01

    Salmonella can contaminate finished products of butcher shops, mainly through cross-contamination of utensils exposed to raw materials. To identify the main sources of contamination with this foodborne pathogen in four butcher shop environments, surface samples were obtained from employees' hands, cutting boards, knives, floor of the refrigeration room, meat grinders, and meat tenderizers (32 samples per area) and analyzed for Salmonella using the International Organization for Standardization method 6579, with modifications. Suspect isolates were identified by PCR (targeting ompC), and confirmed Salmonella isolates were subjected to pulsed-field gel electrophoresis (after treatment with restriction enzyme XbaI), analyzed for the presence of virulence genes (invA, sefA, and spvC), and screened for resistance to 12 antimicrobials. Salmonella isolates was identified only on cutting boards (five samples) from three butcher shops. Fifteen isolates were confirmed as Salmonella belonging to four pulse types (similarity of 71.1 to 100%). The invA gene was detected in 13 isolates, and the sefA was found in 8 isolates; no isolate carried spvC. All tested isolates were resistant to clindamycin and sensitive to amikacin and cefotaxine, and all isolates were resistant to at least 3 of the 12 antimicrobials tested. The results indicate the importance of cutting boards as a source of Salmonella contamination in butcher shops. The presence of multidrug-resistant Salmonella strains possessing virulence genes highlights the health risks for consumers.

  10. Pathotype Classification of Plasmodiophora brassicae Isolates Using Clubroot-Resistant Cultivars of Chinese Cabbage

    Directory of Open Access Journals (Sweden)

    Hun Kim

    2016-10-01

    Full Text Available Clubroot disease caused by Plasmodiophora brassicae is one of the most serious diseases in Brassica crops worldwide. In this study, the pathotypes of 12 Korean P. brassicae field isolates were determined using various Chinese cabbage including 22 commercial cultivars from Korea, China, and Japan, and 15 inbred lines. All P. brassicae isolates exhibited the typical clubroot disease on non-clubroot resistant cultivar, indicating that the isolates were highly pathogenic. According to the reactions on the Williams’ hosts, the 12 field isolates were initially classified into five races. However, when these isolates were inoculated onto clubroot-resistant (CR cultivars of Chinese cabbage, several isolates led to different disease responses even though the isolates have been assigned to the same race by the Williams’ host responses. Based on the pathogenicity results, the 12 field isolates were reclassified into four different groups: pathotype 1 (GN1, GN2, GS, JS, and HS, 2 (DJ and KS, 3 (HN1, PC, and YC, and 4 (HN2 and SS. In addition, the CR cultivars from Korea, China, and Japan exhibited distinguishable disease responses to the P. brassicae isolates, suggesting that the 22 cultivars used in this study, including the non-CR cultivars, are classified into four different host groups based on their disease resistance. Combining these findings, the four differential hosts of Chinese cabbage and four pathotype groups of P. brassicae might provide an efficient screening system for resistant cultivars and a new foundation of breeding strategies for CR Chinese cabbage.

  11. High isoniazid resistance rates in rifampicin susceptible Mycobacterium tuberculosis pulmonary isolates from Pakistan.

    Directory of Open Access Journals (Sweden)

    Naima Fasih

    Full Text Available BACKGROUND: Rapid new diagnostic methods (including Xpert MTB/RIF assay use rifampicin resistance as a surrogate marker for multidrug resistant tuberculosis. Patients infected with rifampicin susceptible strains are prescribed first line anti-tuberculosis therapy. The roll out of such methods raises a concern that strains with resistance to other first line anti-tuberculosis drugs including isoniazid will be missed and inappropriate treatment given. To evaluate implications of using such methods review of resistance data from high burden settings such as ours is essential. OBJECTIVE: To determine resistance to first line anti-tuberculosis drugs amongst rifampicin susceptible pulmonary Mycobacterium tuberculosis (MTB isolates from Pakistan. MATERIALS AND METHODS: Data of pulmonary Mycobacterium tuberculosis strains isolated in Aga Khan University Hospital (AKUH laboratory (2009-2011 was retrospectively analyzed. Antimicrobial susceptibility profile of rifampicin susceptible isolates was evaluated for resistance to isoniazid, pyrazinamide, ethambutol, and streptomycin. RESULTS: Pulmonary specimens submitted to AKUH from 2009 to 2011 yielded 7738 strains of Mycobacterium tuberculosis. These included 54% (n 4183 rifampicin susceptible and 46% (n: 3555 rifampicin resistant strains. Analysis of rifampicin susceptible strains showed resistance to at least one of the first line drugs in 27% (n:1133 of isolates. Overall isoniazid resistance was 15.5% (n: 649, with an isoniazid mono-resistance rate of 4% (n: 174. Combined resistance to isoniazid, pyrazinamide, and ethambutol was noted in 1% (n: 40, while resistance to isoniazid, pyrazinamide, ethambutol, and streptomycin was observed in 1.7% (n: 70 of strains. CONCLUSIONS: Our data suggests that techniques (including Xpert MTB/RIF assay relying on rifampicin susceptibility as an indicator for initiating first line therapy will not detect patients infected with MTB strains resistant to other first line

  12. Dissemination of multidrug-resistant, class 1 integron-carrying Acinetobacter baumannii isolates in Taiwan

    National Research Council Canada - National Science Library

    Huang, L.-Y; Chen, T.-L; Lu, P.-L; Tsai, C.-A; Cho, W.-L; Chang, F.-Y; Fung, C.-P; Siu, L. K

    2008-01-01

    In this study, 283 multidrug-resistant Acinetobacter baumannii (MDR-AB) bloodstream isolates were collected between 1996 and 2004, from three teaching hospitals located in different regions of Taiwan...

  13. OCCURRENCE OF HIGH-LEVEL AMINOGLYCOSIDE RESISTANCE IN ENVIRONMENTAL ISOLATES OF ENTEROCOCCI

    Science.gov (United States)

    High-level resistance fo aminoglycosides was observed in environmental isolates of enterococci. Various aquatic habitats, including agricultural runoff, creeks, rivers, wastewater, and wells, were analyzed. Strains of Enterococcus faecalis, e.faecium, E. gallinarum, and other Ent...

  14. Antimicrobial resistance and virulence of Enterococcus faecalis isolated from retail food

    Science.gov (United States)

    Although enterococci are considered opportunistic nosocomial pathogens, their contribution to food-borne illnesses via dissemination through retail food remains undefined. In this study, prevalence and association of antimicrobial resistance and virulence factors of 80 Enterococcus faecalis isolate...

  15. Antimicrobial Resistance among Campylobacter Isolates Obtained from Retail Chicken Meat and Offal Products in Japan

    National Research Council Canada - National Science Library

    Hidano, Arata; Yamamoto, Takehisa; Hayama, Yoko; Muroga, Norihiko; Kobayashi, Sota; Nishida, Takeshi; Tsutsui, Toshiyuki

    2014-01-01

    .... In this study, we aimed to demonstrate the overall trend in antimicrobial resistance among Campylobacter isolates obtained from chicken meat and offal products collected from a wide geographic area throughout Japan...

  16. Evaluation of DNA microarray for detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis isolates

    Institute of Scientific and Technical Information of China (English)

    王峰

    2013-01-01

    Objective To evaluate the performance of DNA microarray for rapid detection resistance to rifampin and isoniazid in Mycobacterium tuberculosis clinical isolates and identify suitable target sites for molecular genetic test. Methods Twenty-four clinical Mycobacterium

  17. Serotype and Antimicrobial Resistance Patterns of Salmonella Isolates from Commercial Birds and Poultry Environment in Mississippi

    Science.gov (United States)

    To obtain information about Salmonella from commercial birds and poultry environments within Mississippi, 50 Salmonella enterica isolates were collected and characterized by Intergenic Sequence Ribotyping (ISR) serotyping and by determining antimicrobial resistance. ISR assigned serotype to all 50 S...

  18. Arsenic resistance in Campylobacter spp. isolated from retail poultry products.

    Science.gov (United States)

    Sapkota, Amy R; Price, Lance B; Silbergeld, Ellen K; Schwab, Kellogg J

    2006-04-01

    Organoarsenicals are commonly used for growth promotion in U.S. poultry production. Susceptibilities to arsenite, arsenate, and the organoarsenical roxarsone were measured in 251 Campylobacter isolates from conventional and antimicrobial-free retail poultry products. Isolates from conventional poultry products had significantly higher roxarsone MICs (z = 8.22; P < 0.0001).

  19. Antibacterial effect of silver nanoparticles along with protein synthesis-inhibiting antibiotics on Staphylococcus aureus isolated from cattle mastitis

    Directory of Open Access Journals (Sweden)

    Malahat Ahmadi

    2014-01-01

    Full Text Available Introduction: Staphylococcus aureus is an opportunistic pathogen in dairy ruminants which is also found in healthy carriage and can be a major cause of mastitis. Various mastitis control programs have been used to combat the problem but have not always been efficient. In most countries, antibiotic resistance is extremely common. Silver nanoparticles have shown antimicrobial activity against S. aureus. In the present study the effect of silver nanoparticles on S. aureus isolated from cattle mastitis along with antibiotics of operative on protein bacterial synthesis investigated. Materials and methods: Three hundred eleven milk samples were collected from the cow farms. Each milk sample was cultured on mannitol salt agar and was incubated. A total of 72 isolates of S. aureus were isolated from the bovine mastitis milk samples. S. aureus DNA extracted by DNA purification kit according to the manufacturer protocol. 58 isolates were confirmed as S. aureus by biochemical tests as well as nuc gene detection. MIC and MBC determined for silver nanoparticles with antibiotics on 50 isolates. Results: The resistance of S. aureus isolates against erythromycin, gentamicin, streptomycin and doxycycline were 100, 22, 100 and 8%, respectively. 8 of all isolates were sensitive to 25 µg/ml concentration of silver nanoparticles. The 92% growth of the samples were inhibited at concentrations between 50-100 µg/ml. Discussion and conclusion: The present study suggests that antibiotics which can inhibit protein synthesis have significant synergistic effect along with silver nanoparticles.

  20. Phenotypic and genotypic antimicrobial resistance traits of foodborne Staphylococcus aureus isolates from Shanghai.

    Science.gov (United States)

    Xu, Jie; Shi, Chunlei; Song, Minghui; Xu, Xuebin; Yang, Puyu; Paoli, George; Shi, Xianming

    2014-04-01

    Staphylococcus aureus is a recognized pathogen in humans, which causes nosocomial infections and food poisoning. The transmission of antibiotic resistant S. aureus (ARSA), especially methicillin-resistant S. aureus, between food products and humans has become a serious problem. Hence, it is necessary to monitor S. aureus through the food supply chain. In this study, the disk diffusion method and epsilometer test were performed to determine the prevalence of ARSA in 78 foodborne isolates using 18 antibiotics. The highest resistance frequency was found for penicillin G (74.4%), followed by erythromycin (59.0%) and clindamycin (44.9%), whereas no vancomycin-resistant isolates were found. The 78 isolates could be subtyped into 31 resistance profiles and 11 clusters based on their antimicrobial susceptibility. Furthermore, Polymerase chain reaction (PCR) screening for the presence of 13 genes conferring antibiotic resistance was conducted. The presence of resistance genes was relatively high: blaTEM (80.8%), ermB (41.0%), grlA (38.5%), ermC (35.9%), and aac6'/aph2" (35.9%). The incidence of antibiotic resistance was significantly correlated to food types (p = 0.018), with isolates from meat and raw milk more resistant to antibiotics than those from frozen food and vegetables.

  1. Multiple drug resistant carbapenemases producing Acinetobacter baumannii isolates harbours multiple R-plasmids

    Directory of Open Access Journals (Sweden)

    Rajagopalan Saranathan

    2014-01-01

    Full Text Available Background & objectives: The nosocomial human pathogen Acinetobacter baumannii has high propensity to develop resistance to antimicrobials and to become multidrug resistant (MDR, consequently complicating the treatment. This study was carried out to investigate the presence of resistant plasmids (R-plasmids among the clinical isolates of A. baumannii. In addition, the study was performed to check the presence of common β-lactamases encoding genes on these plasmids. Methods: A total of 55 clinical isolates of A. baumannii were included in the study and all were subjected to plasmid DNA isolation, followed by PCR to check the presence of resistance gene determinants such as blaOXA-23 , blaOXA-51, blaOXA-58 and blaIMP-1 on these plasmids that encode for oxacillinase (OXA and metallo-β-lactamase (MBL type of carbapenemases. Plasmid curing experiments were carried out on selected isolates using ethidium bromide and acridine orange as curing agents and the antibiotic resistance profiles were evaluated before and after curing. Results: All the isolates were identified as A. baumannii by 16SrDNA amplification and sequencing. Plasmid DNA isolated from these isolates showed the occurrence of multiple plasmids with size ranging from 500bp to ≥ 25 kb. The percentage of blaOXA-51 and blaOXA-23 on plasmids were found to be 78 and 42 per cent, respectively and 20 isolates (36% carried blaIMP-1 gene on plasmids. Significant difference was observed in the antibiograms of plasmid cured isolates when compared to their parental ones. The clinical isolates became susceptible to more than two antibiotic classes after curing of plasmids indicating plasmid borne resistance. Interpretation & conclusions: Our study determined the plasmid mediated resistance mechanisms and occurrence of different resistance genes on various plasmids isolated from MDR A. baumannii. The present findings showed the evidence for antibiotic resistance mediated through multiple plasmids in

  2. Household Risk Factors for Colonization with Multidrug-Resistant Staphylococcus aureus Isolates

    Science.gov (United States)

    Davis, Meghan F.; Peterson, Amy E.; Julian, Kathleen G.; Greene, Wallace H.; Price, Lance B.; Nelson, Kenrad; Whitener, Cynthia J.; Silbergeld, Ellen K.

    2013-01-01

    Antimicrobial resistance, particularly in pathogens such as methicillin-resistant Staphylococcus aureus (MRSA), limits treatment options and increases healthcare costs. To understand patient risk factors, including household and animal contact, potentially associated with colonization with multidrug-resistant MRSA isolates, we performed a prospective study of case patients colonized with MRSA on admission to a rural tertiary care hospital. Patients were interviewed and antimicrobial resistance patterns were tested among isolates from admitted patients colonized with MRSA in 2009–10. Prevalence of resistance was compared by case-patient risk factors and length-of-stay outcome among 88 MRSA case patients. Results were compared to NHANES 2003–04. Overall prevalence of multidrug resistance (non-susceptibility to ≥four antimicrobial classes) in MRSA nasal isolates was high (73%) and was associated with a 1.5-day increase in subsequent length of stay (p = 0.008). History of hospitalization within the past six months, but not antimicrobial use in the same time period, was associated with resistance patterns. Within a subset of working-age case patients without recent history of hospitalization, animal contact was potentially associated with multidrug resistance. History of hospitalization, older age, and small household size were associated with multidrug resistance in NHANES data. In conclusion, recent hospitalization of case patients was predictive of antimicrobial resistance in MRSA isolates, but novel risk factors associated with the household may be emerging in CA-MRSA case patients. Understanding drivers of antimicrobial resistance in MRSA isolates is important to hospital infection control efforts, relevant to patient outcomes and to indicators of the economic burden of antimicrobial resistance. PMID:23359808

  3. Analysis of Protein Import into Chloroplasts Isolated from Stressed Plants.

    Science.gov (United States)

    Ling, Qihua; Jarvis, Paul

    2016-11-01

    Chloroplasts are organelles with many vital roles in plants, which include not only photosynthesis but numerous other metabolic and signaling functions. Furthermore, chloroplasts are critical for plant responses to various abiotic stresses, such as salinity and osmotic stresses. A chloroplast may contain up to ~3,000 different proteins, some of which are encoded by its own genome. However, the majority of chloroplast proteins are encoded in the nucleus and synthesized in the cytosol, and these proteins need to be imported into the chloroplast through translocons at the chloroplast envelope membranes. Recent studies have shown that the chloroplast protein import can be actively regulated by stress. To biochemically investigate such regulation of protein import under stress conditions, we developed the method described here as a quick and straightforward procedure that can easily be achieved in any laboratory. In this method, plants are grown under normal conditions and then exposed to stress conditions in liquid culture. Plant material is collected, and chloroplasts are then released by homogenization. The crude homogenate is separated by density gradient centrifugation, enabling isolation of the intact chloroplasts. Chloroplast yield is assessed by counting, and chloroplast intactness is checked under a microscope. For the protein import assays, purified chloroplasts are incubated with (35)S radiolabeled in vitro translated precursor proteins, and time-course experiments are conducted to enable comparisons of import rates between genotypes under stress conditions. We present data generated using this method which show that the rate of protein import into chloroplasts from a regulatory mutant is specifically altered under osmotic stress conditions.

  4. Virulence and antimicrobial resistance of Escherichia coli isolated from Tigris River and children diarrhea

    Directory of Open Access Journals (Sweden)

    Ibrahim IA

    2014-11-01

    Full Text Available Israa AJ Ibrahim, Rana M Al-Shwaikh, Mahmoud I IsmaeilDepartment of Biology, College of Education for Pure Science, Ibn Al-Haitham, University of Baghdad, Baghdad, IraqObjective: To investigate the virulence factors including hemolysin production, β-lactamase production, and biofilm formation. Antimicrobial resistance and plasmid content of 20 Escherichia coli isolates obtained from feces and Tigris water were screened.Methods: Ten clinical and ten environmental E. coli isolates were collected from children diarrhea and swim areas on Tigris River in Baghdad city, Iraq, respectively. The bacterial isolates were identified by cultural characteristics, Gram stain, biochemical tests, and screened for the presence of E. coli O157:H7 serotype. Bacterial E. coli isolates were investigated for hemolysin production, biofilm formation, and β-lactamase production. Antibiotics susceptibility and plasmid content were determined.Results: A total of ten clinical and ten water E. coli isolates were studied. Results showed that all E. coli isolates give negative results for latex O157:H7. Virulence factors analysis showed that 6/10 water isolates and 2/10 clinical isolates were hemolytic, 5/10 water isolates and 3/10 clinical isolates were biofilm formation, and 7/10 water isolates and 4/10 clinical isolates were β-lactamase producer. Antibiotics profile showed that all bacterial isolates were multidrug resistant. All E. coli isolates (100% were resistant to carbenicillin, cefodizime, imipenem, and piperacillin. The plasmid DNA analysis showed that all E. coli isolates contained plasmid with molecular weight range between 4.507 kbp and 5.07 kbp, but clinical isolates contained multiple small and mega plasmids.Conclusion: Our study revealed that E. coli isolates from river water exhibit a higher level of hemolysin production, β-lactamase production, and biofilm formation than feces isolates may be due to long adaptation. On the other hand, clinical E. coli

  5. Associations between Antimicrobial Resistance Phenotypes, Antimicrobial Resistance Genes, and Virulence Genes of Fecal Escherichia coli Isolates from Healthy Grow-Finish Pigs ▿

    OpenAIRE

    2009-01-01

    Escherichia coli often carries linked antimicrobial resistance genes on transmissible genetic elements. Through coselection, antimicrobial use may select for unrelated but linked resistance or virulence genes. This study used unconditional statistical associations to investigate the relationships between antimicrobial resistance phenotypes and antimicrobial resistance genes in 151 E. coli isolates from healthy pigs. Phenotypic resistance to each drug was significantly associated with phenotyp...

  6. In vivo development of carbapenem resistance in clinical isolates of Enterobacter aerogenes producing multiple beta-lactamases.

    Science.gov (United States)

    Chen, Ya-Gang; Zhang, Ying; Yu, Yun-Song; Qu, Ting-Ting; Wei, Ze-Qing; Shen, Ping; Li, Lan-Juan

    2008-10-01

    Four clinical strains of extended-spectrum beta-lactamase- and AmpC-producing Enterobacter aerogenes were isolated successively from a liver transplantation patient. Isolates C(1) and C(2) were isolated prior to carbapenem therapy, whilst isolates C(3) and C(4) were recovered after 40 days of carbapenem therapy. The homology of these strains was analysed by pulsed-field gel electrophoresis (PFGE). beta-Lactamases were analysed by isoelectric focusing, polymerase chain reaction (PCR) and sequencing. Outer membrane proteins were analysed by PCR, sequencing, sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blot. Disruption of OmpE36 in C(1) in vitro was also performed by homologous gene recombination. The isolates demonstrated an indistinguishable PFGE pattern. Molecular characterisation revealed that, in addition to the pre-existing multiple beta-lactamases (DHA-1, TEM-1, SHV-5, CTX-M-3 and CTX-M-14) found in C(1) and C(2), isolates C(3) and C(4) failed to express OmpE36 owing to insertional inactivation by an IS903-like insertion sequence. Other resistance mechanisms, such as production of carbapenem-hydrolysing enzymes or expression of chromosomal efflux, were apparently not involved. Completely replacing OmpE36 by the kanamycin resistance gene (kan) resulted in a significant increase in carbapenem minimum inhibitory concentrations of an ompE36 mutant. Thus, C(3) and C(4) were apparently derived from the previously imipenem-susceptible isolates C(1) and C(2). Following carbapenem exposure, depletion of OmpE36 expression resulted in the collateral effect of carbapenem resistance.

  7. [Antimicrobial resistance and molecular epidemiology of Staphylococcus pseudintermedius strains isolated from dog clinical samples].

    Science.gov (United States)

    Vigo, Germán B; Giacoboni, Gabriela I; Gagetti, Paula S; Pasterán, Fernando G; Corso, Alejandra C

    2015-01-01

    Twenty-eight strains isolated from dog clinical samples identified as Staphylococcus pseudintermedius by mass spectrometry (MALDI-TOF) were studied to assess antimicrobial susceptibility by the diffusion method and clonal relationship by pulsed field gel electrophoresis (PFGE). Methicillin resistance (3/28 isolates; 10,7%) was evaluated by mecA PCR. Fifteen strains (53.6%) were resistant to at least one of the antibiotics tested, and eleven of them (39.3%) showed multiple resistance (3 or more antimicrobial families). Eleven isolates (39.3%) were resistant to erythromycin due to the presence of ribosomal methylase ermB, whereas clindamycin inducible resistance was not detected. Twenty-seven (27) clonal types were differentiated by PFGE, suggesting high clonal diversity. We emphasize that the finding of multiresistant S. psedintermedius strains is an emerging problem to be considered in veterinary diagnostic laboratory treatment of canine infections and in public health settings.

  8. Cadmium resistance and uptake by bacterium, Salmonella enterica 43C, isolated from industrial effluent.

    Science.gov (United States)

    Khan, Zaman; Rehman, Abdul; Hussain, Syed Z; Nisar, Muhammad A; Zulfiqar, Soumble; Shakoori, Abdul R

    2016-12-01

    Cadmium resistant bacterium, isolated from industrial wastewater, was characterized as Salmonella enterica 43C on the basis of biochemical and 16S rRNA ribotyping. It is first ever reported S. enterica 43C bared extreme resistance against heavy metal consortia in order of Pb(2+)>Cd(2+)>As(3+)>Zn(2+)>Cr(6+)>Cu(2+)>Hg(2+). Cd(2+) stress altered growth pattern of the bacterium in time dependent manner. It could remove nearly 57 % Cd(2+) from the medium over a period of 8 days. Kinetic and thermodynamic studies based on various adsorption isotherm models (Langmuir and Freundlich) depicted the Cd(2+) biosorption as spontaneous, feasible and endothermic in nature. Interestingly, the bacterium followed pseudo first order kinetics, making it a good biosorbent for heavy metal ions. The S. enterica 43C Cd(2+) processivity was significantly influenced by temperature, pH, initial Cd(2+) concentration, biomass dosage and co-metal ions. FTIR analysis of the bacterium revealed the active participation of amide and carbonyl moieties in Cd(2+) adsorption confirmed by EDX analysis. Electron micrographs beckoned further surface adsorption and increased bacterial size due to intracellular Cd(2+) accumulation. An overwhelming increase in glutathione and other non-protein thiols levels played a significant role in thriving oxidative stress generated by metal cations. Presence of metallothionein clearly depicted the role of such proteins in bacterial metal resistance mechanism. The present study results clearly declare S. enterica 43C a suitable candidate for green chemistry to bioremediate environmental Cd(2+).

  9. Emergence of colistin-resistant Escherichia coli clinical isolates harboring mcr-1 in Vietnam.

    Science.gov (United States)

    Tada, Tatsuya; Nhung, Pham Hong; Shimada, Kayo; Tsuchiya, Mitsuhiro; Phuong, Doan Mai; Anh, Nguyen Quoc; Ohmagari, Norio; Kirikae, Teruo

    2017-07-10

    The mcr-1 was first detected on a plasmid in colistin-resistant Escherichia coli from livestock and patients in China. We described here the emergence of colistin-resistant E. coli clinical isolates harboring mcr-1 on the chromosomes in Vietnam. To our knowledge, this is the first report of hospital-acquired E. coli isolates harboring mcr-1 in a medical setting in Vietnam. Copyright © 2017. Published by Elsevier Ltd.

  10. Multiple Antimicrobial Resistance of Escherichia coli Isolated from Chickens in Iran

    OpenAIRE

    Reza Talebiyan; Mehdi Kheradmand; Faham Khamesipour; Mohammad Rabiee-Faradonbeh

    2014-01-01

    Antimicrobial agents are used extremely in order to reduce the great losses caused by Escherichia coli infections in poultry industry. In this study, 318 pathogenic Escherichia coli (APEC) strains isolated from commercial broiler flocks with coli-septicemia were examined for antimicrobials of both veterinary and human significance by disc diffusion method. Multiple resistances to antimicrobial agents were observed in all the isolates. Resistance to the antibiotics was as follows: Tylosin (88....

  11. First report of IMI-1-producing colistin-resistant Enterobacter clinical isolate in Ireland, March 2013.

    Science.gov (United States)

    Boo, T W; O'Connell, N; Power, L; O'Connor, M; King, J; McGrath, E; Hill, R; Hopkins, K L; Woodford, N

    2013-08-01

    We report the first case in Ireland of an IMI-1 carbapenemase-producing Enterobacter asburiae, which was resistant to both colistin and fosfomycin. The circumstances under which this isolate was acquired were unclear. Several reports of IMI-producing Enterobacter spp. have emerged in recent years, and colistin resistance in Enterobacteriaceae is also increasingly reported. Laboratories should be aware of the unusual antibiograms of IMI-producing isolates.

  12. pncA Mutations in Pyrazinamide-Resistant Mycobacterium tuberculosis Isolates from Northwestern Russia

    OpenAIRE

    1999-01-01

    Thirty-six pyrazinamide-resistant and eight pyrazinamide-susceptible Mycobacterium tuberculosis isolates from Russia were analyzed for their pncA mutations. Thirty-one (86.1%) of the resistant isolates had a mutation either in pncA or upstream of the gene. Twenty of the 23 different mutations found in this study had not been described earlier. pncA genotype correlated well with pyrazinamidase activity and BACTEC 460 susceptibility test results.

  13. Isolation of Salmonella spp. and drug resistance to tetracycline , ampicillin and cotrimoxazole in kerman

    Directory of Open Access Journals (Sweden)

    Shahrnaz banuo Ashrafganjooyi

    2014-08-01

    Results and Conclusion: In this study 56 Salmonella spp were isolated in this rigion .Drug resistance pattern to antibiotics such as tetracycline , ampicillin and cotrimoxazole were: 55.3%, 7.5%, 35.7% with average MIC respectively: (MIC≥16µg/ml (MIC ≥ 32µg/ml ( MIC ≥ 4.76µg/ml From 56 Salmonella spp which were isolated from this region ,the most resistant was to tetracycline.

  14. Bacillus subtilis vegetative isolate surviving chlorine dioxide exposure: an elusive mechanism of resistance.

    Science.gov (United States)

    Martin, D J H; Wesgate, R L; Denyer, S P; McDonnell, G; Maillard, J-Y

    2015-12-01

    Oxidizing agents such as chlorine dioxide are widely used microbicides, including for disinfection of medical equipment. We isolated a Bacillus subtilis isolate from a washer-disinfector whose vegetative form demonstrated unique resistance to chlorine dioxide (0·03%) and hydrogen peroxide (7·5%). The aim of this study was to understand the mechanisms of resistance expressed by this isolate. A range of resistance mechanisms were investigated in the B. subtilis isolate and a reference B. subtilis strain (ATCC 6051) to include bacterial cell aggregation, the presence of profuse exopolysaccharide (EPS), and the expression of detoxification enzymes. The basis of resistance of the isolate to high concentrations of oxidizing agents was not linked to the presence of endospores. Although, the presence of EPS, aggregation and expression of detoxification enzymes may play a role in bacterial survival to low concentrations of chlorine dioxide, it is unlikely that the mechanisms helped tested to survive the bactericidal effect of higher oxidizer concentrations. Overall, the mechanisms conferring resistance to chlorine dioxide and hydrogen peroxide remains elusive. Based on recent advances in the mode of action of oxidizing agents and notably hydrogen peroxide, we postulate that additional efficient intracellular mechanisms may be involved to explain significant resistance to in-use concentrations of commonly used high-level disinfectants. The isolation of a highly resistant vegetative Gram-positive bacterium to a highly reactive oxidizing agent is worrying. Understanding the mechanisms conferring such resistance is essential to effectively control such bacterial isolates. Here, we postulate that there are still mechanisms of bacterial resistance that have not been fully characterized. © 2015 The Authors published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology.

  15. Prevalence of macrolide-non-susceptible isolates among β-lactamase-negative ampicillin-resistant Haemophilus influenzae in a tertiary care hospital in Japan.

    Science.gov (United States)

    Wajima, Takeaki; Seyama, Shoji; Nakamura, Yuka; Kashima, Chihiro; Nakaminami, Hidemasa; Ushio, Masanobu; Fujii, Takeshi; Noguchi, Norihisa

    2016-09-01

    β-Lactamase-negative ampicillin-resistant (BLNAR) Haemophilus influenzae account for a large portion of H. influenzae clinical isolates in Japan. The aim of this study was to clarify the antimicrobial susceptibility of BLNAR H. influenzae clinical isolates as well as the annual changes in susceptibility. BLNAR H. influenzae isolates were collected from a tertiary care hospital from 2007 to 2012. Antimicrobial susceptibility testing was performed and resistance mechanisms were analysed. All of the isolates (n=304) had amino acid substitutions in penicillin-binding protein 3 (PBP3) and isolates were classified by these amino acid substitutions: R517H or N526K (class I); S385T and R517H (class II); and S385T and N526K (class III). Classes I, II and III represented 8.2% (n=25), 9.5% (n=29) and 81.6% (n=248) of the isolates, respectively; 2 isolates could not be classified because they had a PBP3 with a substantially mutated FtsI transpeptidase domain. All of the isolates were highly susceptible to fluoroquinolones and carbapenems. The number of clarithromycin (CAM)-non-susceptible [minimum inhibitory concentration (MIC) ≥16μg/mL] H. influenzae isolates increased significantly between 2010 and 2012. Moreover, CAM-non-susceptible H. influenzae isolates were prevalent among class II and class III BLNAR H. influenzae. Multilocus sequence typing (MLST) of the CAM-resistant (MIC ≥32μg/mL) H. influenzae isolates showed that they were not specific sequence types, suggesting that CAM resistance may occur in any isolates. These results raise concern regarding the occurrence of multidrug-resistant BLNAR H. influenzae.

  16. Antimicrobial resistance of Listeria monocytogenes isolated from seafood and humans in Iran.

    Science.gov (United States)

    Abdollahzadeh, Esmail; Ojagh, Seyed Mahdi; Hosseini, Hedayat; Ghaemi, Ezzat Allah; Irajian, Gholamreza; Naghizadeh Heidarlo, Masoud

    2016-11-01

    Fourteen Listeria monocytogenes isolates previously collected from seafood (n = 7) and human patients (n = 7) were studied for their antimicrobial susceptibility against eight common antimicrobials (ampicillin, penicillin, gentamicin, streptomycin, tetracycline, trimethoprim-sulfamethoxazole, chloramphenicol, and cefotaxime). A high resistance level to ampicillin, cefotaxime (100%), and pencillin (57% in seafood isolates and 71.4% in clinical isolates) was observed in this study. However, all of the isolates were susceptible to trimethoprim-sulfamethoxazole, chloramphenicol, and tetracycline. Simultaneous resistance was identified in 4 clinical isolates (57.1%). Genotypic characterization of fish isolates (isolated from three fish species) was performed by pulsed-field gel electrophoresis (PFGE). A high diversity among fish isolates was observed. PFGE analyses distinguished the 4 isolates into 4 reproducible pulsotypes. There was no correlation between the antibiograms with pulsotypes. In conclusion, the resistance of seafood isolates to the antibiotics commonly used to treat listeriosis could be a potential health hazard for consumers. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Retinol binding protein 4, obesity, and insulin resistance in adolescents

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    Ronaldi Noor

    2017-02-01

    Full Text Available Background Obesity is a global problem. Even in poor and developing countries, obesity has reached alarming levels. In childhood, obesity may lead to insulin resistance. Retinol binding protein (RBP4, secreted primarily by liver and adipose tissues, was recently proposed as a link between obesity and insulin resistance. The role of RBP4 in pediatric obesity and its relationship with insulin resistance have not been well elucidated. Objective To compare RBP4 levels in obese and lean adolescents and to assess for a relationship between RBP4 levels and insulin resistance. Method This cross-sectional study was conducted in three senior high schools in Padang, West Sumatera, Indonesia. Subjects were adolescents aged 14-18 years, who were obese or normal weight (n=56. We measured subjects’ body mass index (BMI and serum RBP4 concentrations. Insulin resistance was assessed using the homeostasis model assessment of insulin resistance (HOMA-IR index. Results Similar RBP4 levels were found in the obese and normoweight groups (P>0.05. Higher RBP4 levels were found in the insulin resistant compared to the non-insulin resistant group, but the difference was not significant (P > 0.05. Conclusion There is no significant difference in mean RBP4 levels in obese adolescents compared to normoweight adolescents. Nor are mean RBP4 levels significantly different between obese adolescents with and without insulin resistance.

  18. The Bioinformatics Report of Mutation Outcome on NADPH Flavin Oxidoreductase Protein Sequence in Clinical Isolates of H. pylori.

    Science.gov (United States)

    Mirzaei, Nasrin; Poursina, Farkhondeh; Moghim, Sharareh; Ghaempanah, Abdol Majid; Safaei, Hajieh Ghasemian

    2016-05-01

    frxA gene has been implicated in the metronidazole nitro reduction by H. pylori. Alternatively, frxA is expected to contribute to the protection of urease and to the in vivo survival of H. pylori. The aim of present study is to report the mutation effects on the frxA protein sequence in clinical isolates of H. pylori in our community. Metronidazole resistance was proven in 27 of 48 isolates. glmM and frxA genes were used for molecular confirmation of H. pylori isolates. The primer set for detection of whole sequence of frxA gene for the effect of mutation on protein sequence was used. DNA and protein sequence evaluation and analysis were done by blast, Clustal Omega, and T COFFEE programs. Then, FrxA protein sequences from six metronidazole-resistant clinical isolates were analyzed by web-based bioinformatics tools. The result of six metronidazole-resistant clinical isolates in comparison with strain 26695 showed ten missense mutations. The result with the STRING program revealed that no change was seen after alterations in these sequences. According to consensus data involving four methods, residue substitutions at 40, 13, and 141 increase the stability of protein sequence after mutation, while other alterations decrease. Residue substitutions at 40, 43, 141, 138, 169, and 179 are deleterious, while, V7I, Q10R, V34I, and V96I alterations are neutral. As FrxA contribute to survival of bacterium and in regard to the effect of mutations on protein function, it might affect the survival and bacterium phenotype and it need to be studied more. Also, none of the stability prediction tool is perfect; iStable is the best predictor method among all methods.

  19. Primary steps of the photoactive yellow protein: Isolated chromophore dynamics and protein directed function

    OpenAIRE

    Lee, I-Ren; Lee, Wonchul; Zewail, Ahmed H.

    2006-01-01

    The cycle of the photoactive yellow protein (PYP) has been extensively studied, but the dynamics of the isolated chromophore responsible for transduction is unknown. Here, we present real-time observation of the dynamics of the negatively charged chromophore and detection of intermediates along the path of trans-to-cis isomerization using femtosecond mass selection/electron detachment techniques. The results show that the role of the protein environment is not in the first step of double-bond...

  20. Detection and prevalence of antimicrobial resistance genes in Campylobacter spp. isolated from chickens and humans

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    Samantha Reddy

    2017-01-01

    Full Text Available Campylobacter spp. are common pathogenic bacteria in both veterinary and human medicine. Infections caused by Campylobacter spp. are usually treated using antibiotics. However, the injudicious use of antibiotics has been proven to spearhead the emergence of antibiotic resistance. The purpose of this study was to detect the prevalence of antibiotic resistance genes in Campylobacter spp. isolated from chickens and human clinical cases in South Africa. One hundred and sixty one isolates of Campylobacter jejuni and Campylobacter coli were collected from chickens and human clinical cases and then screened for the presence of antimicrobial resistance genes. We observed a wide distribution of the tetO gene, which confers resistance to tetracycline. The gyrA genes that are responsible quinolone resistance were also detected. Finally, our study also detected the presence of the blaOXA-61, which is associated with ampicillin resistance. There was a higher (p < 0.05 prevalence of the studied antimicrobial resistance genes in chicken faeces compared with human clinical isolates. The tetO gene was the most prevalent gene detected, which was isolated at 64% and 68% from human and chicken isolates, respectively. The presence of gyrA genes was significantly (p < 0.05 associated with quinolone resistance. In conclusion, this study demonstrated the presence of gyrA (235 bp, gyrA (270 bp, blaOXA-61 and tetO antimicrobial resistance genes in C. jejuni and C. coli isolated from chickens and human clinical cases. This indicates that Campylobacter spp. have the potential of resistance to a number of antibiotic classes.

  1. A Survey of the Epidemiology and Antibiotic Resistance Patterns of Enteropathogens Isolates in an Iranian Hospital

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    Mirjafari Tafti

    2016-02-01

    Full Text Available Background Infectious diarrhea is one of the most frequent diseases among children, especially in developing countries. Objectives The aim of this study was to determine the etiological agents and drug resistance patterns of common enteric pathogens isolated in an Iranian 1000-bed tertiary care hospital. Patients and Methods In a retrospective study, we analyzed the etiology and drug resistance patterns of enteric pathogens associated with diarrheal cases. The study was carried out in the Milad hospital of Tehran over two years, from April of 2012 to January of 2014. Stool specimens from patients with diarrhea (n = 7321 were examined for enteric pathogens using routine microbiological culture methods. Strains of Salmonella, Shigella, and enteric pathogenic E. coli (EPEC were serotyped and their susceptibility to commonly used antimicrobial agents was determined by a disk diffusion method, as recommended by the clinical and laboratory standards institute (CLSI guidelines. Results Enteric pathogens were isolated from 310 (4.23% of the patients. The most frequently isolated microorganisms included enteropathogenic E. coli (EPEC, Salmonella, and Shigella spp. The majority of the isolates of EPEC were resistant to commonly used antibiotics such as ampicillin (85.61%, cefixime (79.41%, and nalidixic acid. Resistance among other enteric pathogens was also prevalent. About 45.70% of the Salmonella isolates were resistant to chloramphenicol, and 87.95% were resistant to sulfamethoxazole/trimethoprim. Resistance of the Shigella isolates to nalidixic acid in comparison to the resistance recorded in previous studies was higher. Conclusions The results show that enteric bacteria, including EPEC, Salmonella spp., and Shigella spp. are the major causative agents of diarrhea in the hospital. The emergence of antimicrobial resistance among enteric pathogens is an important problem for public health. Considering the threat of emerging antimicrobial resistance

  2. The resistance of Salmonella spp. isolates from Alabio duck against several antibiotics

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    Istiana

    1998-06-01

    Full Text Available A total of 172 isolates of Salmonella spp. consisted of S. typhimurium (70, S. Radar (52, S. senftenberg (25, S. Virchow (14, and S. amsterdam (11 from Alabio duck hatcheries in the District of Hulu Sungai Utara, South Kalimantan, were examined their resistencies against seven kinds of antibiotics, consisted of chloramphenicol, neomycin, trimethoprim, streptomycin, tetracycline, ampicillin, and polymixin B, by using agar disc diffusion method. The results showed that 70 isolates of S. typhimurium were resistant against six kinds of antibiotics with various percentages, that is chlorampenicol 5 .7%, neomycin 12 .8%, trimethoprim 7.1 %, streptomycin 8 .6%, tetracycline 11 .4%, and the highest against ampicillin 30 %. Ofthe 52 S. hadar isolates showed that all ofthem were resistent against 6 kinds of antibiotics, that is chloramphenicol 1 .9%, neomycin 7.7%, trimethoprim 5.8%, streptomycin 48 .0%, tetracycline 48.0%, and polymixin B 3.8%. Futhermore S. senftenberg (25 isolates were resistent against three kinds of antibiotics, that is neomycin 12%, streptomycin 20%, and tetracyclines 16%. Meanwhile S. virchow (14 isolates, were resistant against two kinds of antibiotics namely streptomycin 7.1% and tetracylines 14.3%. Whereas S. amsterdam (11 isolates were resistant against four kinds of antibiotics, that is neomycin 45 .5%, streptomycin 18 .2%, tetracycline 18 .2%, and ampicillin 9.10%.

  3. Resistive random access memory utilizing ferritin protein with Pt nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Uenuma, Mutsunori; Kawano, Kentaro; Zheng Bin; Okamoto, Naofumi; Horita, Masahiro; Yoshii, Shigeo; Yamashita, Ichiro; Uraoka, Yukiharu, E-mail: uenuma@ms.naist.jp [Nara Institute of Science and Technology, 8916-5, Takayama, Ikoma, Nara 630-0192 (Japan)

    2011-05-27

    This study reports controlled single conductive paths found in resistive random access memory (ReRAM) formed by embedding Pt nanoparticles (Pt NPs) in NiO film. Homogeneous Pt NPs produced and placed by ferritin protein produce electric field convergence which leads to controlled conductive path formation. The ReRAM with Pt NPs shows stable switching behavior. A Pt NP density decrease results in an increase of OFF state resistance and decrease of forming voltage, whereas ON resistance was independent of the Pt NP density, which indicates that a single metal NP in a memory cell will achieve low power and stable operation.

  4. Antimicrobial resistance of non-typhoidal Salmonella isolates from egg layer flocks and egg shells.

    Science.gov (United States)

    Pande, Vivek V; Gole, Vaibhav C; McWhorter, Andrea R; Abraham, Sam; Chousalkar, Kapil K

    2015-06-16

    This study was conducted to examine the antimicrobial resistance (AMR) of Salmonella spp. isolated from commercial caged layer flocks in New South Wales and South Australia. All Salmonella isolates (n=145) were subjected to phenotypic and genotypic characterisation of AMR and carriage of integrons. The majority of Salmonella isolates (91.72%) were susceptible to all antimicrobials tested in this study. Limited resistance was observed to amoxicillin and ampicillin (5.51%), tetracycline (4.13%), cephalothin (2.06%) and trimethoprim (0.68%). None of the isolates were resistant to cefotaxime, ceftiofur, ciprofloxacin, chloramphenicol, gentamycin, neomycin or streptomycin. A low frequency of Salmonella isolates (4.83%) harboured antimicrobial resistance genes and a class 1 integron. The most commonly detected AMR genes among the Salmonella isolates were blaTEM (2.07%), tet A (1.38%) and dhfrV (0.69%). Overall, Salmonella enterica isolates exhibited a low frequency of AMR and represent a minimal public health risk associated with the emergence of multidrug resistant Salmonella spp. from the Australian layer industry.

  5. Synergistic effects of resistance training and protein intake: practical aspects.

    Science.gov (United States)

    Guimarães-Ferreira, Lucas; Cholewa, Jason Michael; Naimo, Marshall Alan; Zhi, X I A; Magagnin, Daiane; de Sá, Rafaele Bis Dal Ponte; Streck, Emilio Luiz; Teixeira, Tamiris da Silva; Zanchi, Nelo Eidy

    2014-10-01

    Resistance training is a potent stimulus to increase skeletal muscle mass. The muscle protein accretion process depends on a robust synergistic action between protein intake and overload. The intake of protein after resistance training increases plasma amino acids, which results in the activation of signaling molecules leading to increased muscle protein synthesis (MPS) and muscle hypertrophy. Although both essential and non-essential amino acids are necessary for hypertrophy, the intake of free L-leucine or high-leucine whole proteins has been specifically shown to increase the initiation of translation that is essential for elevated MPS. The literature supports the use of protein intake following resistance-training sessions to enhance MPS; however, less understood are the effects of different protein sources and timing protocols on MPS. The sum of the adaptions from each individual training session is essential to muscle hypertrophy, and thus highlights the importance of an optimal supplementation protocol. The aim of this review is to present recent findings reported in the literature and to discuss the practical application of these results. In that light, new speculations and questions will arise that may direct future investigations. The information and recommendations generated in this review should be of benefit to clinical dietitians as well as those engaged in sports.

  6. Tunable protein-resistance of polycation-terminated polyelectrolyte multilayers.

    Science.gov (United States)

    Tristán, Ferdinando; Palestino, Gabriela; Menchaca, J-Luis; Pérez, Elías; Atmani, Hassan; Cuisinier, Frédéric; Ladam, Guy

    2009-08-10

    The prevention of nonspecific protein adsorption is a crucial prerequisite for many biomedical and biotechnological applications. Therefore, the design of robust and versatile methods conferring optimal protein-resistance properties to surfaces has become a challenging issue. Here we report the unexpected case of polycation-ending polyelectrolyte multilayers (PEM) that efficiently prevented the adsorption of a negatively charged model protein, glucose oxidase (GOX). PEM films were based on two typical weak poyelectrolytes: poly(allylamine hydrochloride) (PAH) and poly(acrylic acid) (PAA). No chemical modification of the polyelectrolytes was required and tunable GOX adsorption was possible by simply changing the buildup pH conditions. Protein-resistance properties are attributed to high film hydration becoming the predominant factor over electrostatic interactions. We explain this effect by oscillations of the internal PAA ionization state throughout the buildup, which results in an excess of carboxylic acid groups within the film. This excess acts as a reservoir of potential carboxylate groups compensating the outer PAH positive charges. Partial results indicated that the system was also resistant to the adsorption of a positively charged protein, lysozyme. Control of the internal ionization of weak polyelectrolyte multilayers might open a route toward simple tuning of protein adsorption. These results should help to rationalize the design of biomaterials, biosensors, or protein separation devices.

  7. Drug resistance, plasmids, biotypes and susceptibility to bacteriophages of Salmonella isolated from poultry in Canada.

    Science.gov (United States)

    Poppe, C; McFadden, K A; Demczuk, W H

    1996-07-01

    Salmonella isolates from 295 layer and 294 broiler flocks in Canada were examined to determine resistance to antimicrobial agents, plasmid profiles, biochemical properties, and susceptibility to polyvalent bacteriophages. Except for the high number of strains resistant to spectinomycin (97.8%), the frequency of drug resistance of Salmonella isolates from layer flocks was low. None of 457 isolates from layer flocks was resistant to amikacin or ciprofloxacin, and less than 2% of the strains were resistant to cephalothin, chloramphenicol, cotrimoxazole, gentamicin, kanamycin, neomycin, nitrofurantoin, and/or polymyxin B. About 3% of the strains were resistant to ampicillin, carbenicillin and/or tetracycline, whereas 8% of the strains were resistant to sulfisoxazole. Salmonella anatum var. O15+ and S. typhimurium var. copenhagen strains were resistant to multiple antimicrobial agents. None of 1159 Salmonella strains from broiler flocks was resistant to amikacin, cephalothin, ciprofloxacin or polymyxin B, less than 1% of the strains were resistant to chloramphenicol, 2% were resistant to ampicillin, carbenicillin and/or chloramphenicol; 5-7% were resistant to the aminoglycosides gentamicin, kanamycin and/or neomycin; 6% were resistant to nitrofurantoin; 10% to tetracycline; 14% to sulfisoxazole; and 99% to spectinomycin. A high percentage of S. binza, S. anatum var. O15+, S. schwarzengrund and S. heidelberg strains were resistant to antimicrobial agents. Some of the single or multiple resistances were encoded by conjugative plasmids or by plasmids that were thermosensitive for transfer. Eight percent of S. heidelberg strains did not produce hydrogen sulfide. Ninety-seven percent of the Salmonella strains were susceptible to the lytic effect of polyvalent bacteriophages.

  8. Antimicrobial resistance and presence of the SXT mobile element in Vibrio spp. isolated from aquaculture facilities.

    Science.gov (United States)

    García-Aljaro, Cristina; Riera-Heredia, Jordi; Blanch, Anicet R

    2014-07-01

    The aim of this work was to assess the susceptibility of Vibrio spp. strains isolated from fish cultures against some usually applied antibiotics and the occurrence of the SXT mobile genetic element among them. Antimicrobial resistance was assessed by the standard disk diffusion technique while the presence of the SXT mobile genetic element was determined by conventional PCR. High levels of resistance to ampicillin (70%), cefoxitin (44%), streptomycin (31%), aztreonam (25%) and sulfamethoxazole (21%) were detected, and a high inter-and-intraspecies diversity in the resistance profile was observed for the majority of the analysed isolates. The SXT mobile genetic element was detected in only 4 isolates belonging to the species V. diazotrophicus (1), V. mediterranei (2) and V. vulnificus (1), which showed a variable antibiotic resistance profile. Horizontal antibiotic resistance gene transfer from the V. diazotrophicus SXT-positive strain to a laboratory E. coli strain was demonstrated under laboratory conditions. Our results suggest that the Vibrio spp. isolated from aquaculture facilities analysed in this study, although not being pathogenic, they constitute a source of antimicrobial resistance genes that could be mobilized to other bacterial populations through mobile genetic elements. However, the low occurrence of the SXT element in these isolates supports the hypothesis that this element is not involved in the development of resistance in the majority of Vibrio spp. in the examined aquaculture facilities.

  9. Detection of Methicillin-Resistance Gene in Staphylococcus aureus Isolated from Traditional White Cheese in Iran

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    Mina Varmazyar-najafi

    2016-09-01

    Full Text Available Background & Aims of the Study: Methicillin-resistant staphylococcus aureus (MRSA is considered as a major pathogen in public health concern. The objectives of this study were to firstly determine antibiotic sensitivity among Staphylococcus aureus isolated from traditional Iranian white cheese during 2015 from Hamedan province of Iran; and secondly to estimate the presence of methicillin-resistant S. aureus. Materials &Methods: This cross-sectional study was done by collecting 120 Iranian white cheeses (traditional and industrial which were available in different markets; and tested for the presence of S. aureus by culture methods. The obtained isolates were subjected to disc diffusion antimicrobial susceptibility tests followed by PCR detection of the mecA gene. Results: Out of 120 examined cheese samples, 19 samples (31.67% were contaminated with S. aureus. The highest rate of antibiotic resistance was observed for penicillin, as all of the 19 isolates (100% were found to be resistant to this antibiotic using disk diffusion method. Three out of 19 S. aureus isolates (15.7% were phenotypically resistant to methicillin (disk diffusion, while 4 (21.05% of them were genotypically confirmed as MRSA strains. Furthermore, none of the isolates were found resistant to vancomycin. Conclusion: The results of the study confirm the presence of methicillin resistant strains of S. aureus in Iranian white cheese. It should be considered to constitute a potential health risk for consumers, suggesting usage of more stringent hygiene measures.

  10. Low rate of fluoroquinolone resistance in Mycobacterium tuberculosis isolates from northern Tanzania.

    Science.gov (United States)

    van den Boogaard, Jossy; Semvua, Hadija H; van Ingen, Jakko; Mwaigwisya, Solomon; van der Laan, Tridia; van Soolingen, Dick; Kibiki, Gibson S; Boeree, Martin J; Aarnoutse, Rob E

    2011-08-01

    Fluoroquinolones are used in second-line treatment of tuberculosis (TB) and have a potential role in shortening TB treatment duration. The wide use of fluoroquinolones in the treatment of other infections, including respiratory tract infections in patients with (undiagnosed) active TB, could result in fluoroquinolone-resistant Mycobacterium tuberculosis. We determined the rate of fluoroquinolone resistance in M. tuberculosis isolates obtained from Tanzanian patients and linked this to previous fluoroquinolone exposure and mycobacterial resistance to rifampicin and isoniazid. A total of 291 M. tuberculosis isolates were obtained between April 2009 and June 2010 from patients with smear-positive pulmonary TB and tested for susceptibility to ciprofloxacin, moxifloxacin, rifampicin and isoniazid. Information on previous fluoroquinolone use was obtained by interviewing patients and checking their medical files. Only 2 (0.7%) of the 291 M. tuberculosis isolates were resistant to ciprofloxacin; 1 of which was intermediately resistant to moxifloxacin as well. These two isolates were susceptible to rifampicin and isoniazid. Twenty-two (8%) of the 291 patients had a history of fluoroquinolone use (median: 7 days; interquartile range: 5-10 days). The patients from whom the fluoroquinolone-resistant M. tuberculosis isolates were obtained had no known history of previous fluoroquinolone use. Our findings indicate that the rate of fluoroquinolone-resistant M. tuberculosis in Tanzanian patients with TB is low and not related to previous, brief episodes of exposure to fluoroquinolones. The findings favour future application of fluoroquinolones in TB treatment regimens of shorter duration.

  11. Antibiotic resistance determinants in a Pseudomonas putida strain isolated from a hospital.

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    Lázaro Molina

    Full Text Available Environmental microbes harbor an enormous pool of antibiotic and biocide resistance genes that can impact the resistance profiles of animal and human pathogens via horizontal gene transfer. Pseudomonas putida strains are ubiquitous in soil and water but have been seldom isolated from humans. We have established a collection of P. putida strains isolated from in-patients in different hospitals in France. One of the isolated strains (HB3267 kills insects and is resistant to the majority of the antibiotics used in laboratories and hospitals, including aminoglycosides, ß-lactams, cationic peptides, chromoprotein enediyne antibiotics, dihydrofolate reductase inhibitors, fluoroquinolones and quinolones, glycopeptide antibiotics, macrolides, polyketides and sulfonamides. Similar to other P. putida clinical isolates the strain was sensitive to amikacin. To shed light on the broad pattern of antibiotic resistance, which is rarely found in clinical isolates of this species, the genome of this strain was sequenced and analysed. The study revealed that the determinants of multiple resistance are both chromosomally-borne as well as located on the pPC9 plasmid. Further analysis indicated that pPC9 has recruited antibiotic and biocide resistance genes from environmental microorganisms as well as from opportunistic and true human pathogens. The pPC9 plasmid is not self-transmissible, but can be mobilized by other bacterial plasmids making it capable of spreading antibiotic resistant determinants to new hosts.

  12. Antibiotic Resistance of Salmonella spp. Isolated from Shrimp Farming Freshwater Environment in Northeast Region of Brazil

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    Fátima C. T. Carvalho

    2013-01-01

    Full Text Available This study investigated the presence and antibiotic resistance of Salmonella spp. in a shrimp farming environment in Northeast Region of Brazil. Samples of water and sediments from two farms rearing freshwater-acclimated Litopenaeus vannamei were examined for the presence of Salmonella. Afterwards, Salmonella isolates were serotyped, the antimicrobial resistance was determined by a disk diffusion method, and the plasmid curing was performed for resistant isolates. A total of 30 (16.12% of the 186 isolates were confirmed to be Salmonella spp., belonging to five serovars: S. serovar Saintpaul, S. serovar Infantis, S. serovar Panama, S. serovar Madelia, and S. serovar Braenderup, along with 2 subspecies: S. enterica serovar houtenae and S. enterica serovar enterica. About twenty-three percent of the isolates were resistant to at least one antibiotic, and twenty percent were resistant to at least two antibiotics. Three strains isolated from water samples (pond and inlet canal exhibited multiresistance to ampicillin, tetracycline, oxytetracycline, and nitrofurantoin. One of them had a plasmid with genes conferring resistance to nitrofurantoin and ampicillin. The incidence of bacteria pathogenic to humans in a shrimp farming environment, as well as their drug-resistance pattern revealed in this study, emphasizes the need for a more rigorous attention to this area.

  13. Antibiotic Resistance of Salmonella spp. Isolated from Shrimp Farming Freshwater Environment in Northeast Region of Brazil

    Science.gov (United States)

    Carvalho, Fátima C. T.; Sousa, Oscarina V.; Carvalho, Edirsana M. R.; Hofer, Ernesto; Vieira, Regine H. S. F.

    2013-01-01

    This study investigated the presence and antibiotic resistance of Salmonella spp. in a shrimp farming environment in Northeast Region of Brazil. Samples of water and sediments from two farms rearing freshwater-acclimated Litopenaeus vannamei were examined for the presence of Salmonella. Afterwards, Salmonella isolates were serotyped, the antimicrobial resistance was determined by a disk diffusion method, and the plasmid curing was performed for resistant isolates. A total of 30 (16.12%) of the 186 isolates were confirmed to be Salmonella spp., belonging to five serovars: S. serovar Saintpaul, S. serovar Infantis, S. serovar Panama, S. serovar Madelia, and S. serovar Braenderup, along with 2 subspecies: S. enterica serovar houtenae and S. enterica serovar enterica. About twenty-three percent of the isolates were resistant to at least one antibiotic, and twenty percent were resistant to at least two antibiotics. Three strains isolated from water samples (pond and inlet canal) exhibited multiresistance to ampicillin, tetracycline, oxytetracycline, and nitrofurantoin. One of them had a plasmid with genes conferring resistance to nitrofurantoin and ampicillin. The incidence of bacteria pathogenic to humans in a shrimp farming environment, as well as their drug-resistance pattern revealed in this study, emphasizes the need for a more rigorous attention to this area. PMID:24455280

  14. Functional characterization of bacteria isolated from ancient arctic soil exposes diverse resistance mechanisms to modern antibiotics.

    Science.gov (United States)

    Perron, Gabriel G; Whyte, Lyle; Turnbaugh, Peter J; Goordial, Jacqueline; Hanage, William P; Dantas, Gautam; Desai, Michael M

    2015-01-01

    Using functional metagenomics to study the resistomes of bacterial communities isolated from different layers of the Canadian high Arctic permafrost, we show that microbial communities harbored diverse resistance mechanisms at least 5,000 years ago. Among bacteria sampled from the ancient layers of a permafrost core, we isolated eight genes conferring clinical levels of resistance against aminoglycoside, β-lactam and tetracycline antibiotics that are naturally produced by microorganisms. Among these resistance genes, four also conferred resistance against amikacin, a modern semi-synthetic antibiotic that does not naturally occur in microorganisms. In bacteria sampled from the overlaying active layer, we isolated ten different genes conferring resistance to all six antibiotics tested in this study, including aminoglycoside, β-lactam and tetracycline variants that are naturally produced by microorganisms as well as semi-synthetic variants produced in the laboratory. On average, we found that resistance genes found in permafrost bacteria conferred lower levels of resistance against clinically relevant antibiotics than resistance genes sampled from the active layer. Our results demonstrate that antibiotic resistance genes were functionally diverse prior to the anthropogenic use of antibiotics, contributing to the evolution of natural reservoirs of resistance genes.

  15. Detection of benzalkonium chloride resistance in community environmental isolates of staphylococci.

    Science.gov (United States)

    He, Gui-Xin; Landry, Michael; Chen, Huizhong; Thorpe, Conner; Walsh, Dennis; Varela, Manuel F; Pan, Hongmiao

    2014-05-01

    We isolated a total of 653 strains from 64 community environmental samples in Massachusetts, USA. Among these isolates, 9.65 % (63 strains) were benzalkonium chloride (BC)-resistant staphylococci. All BC-resistant strains were collected from surfaces upon which antibacterial wipes or antibacterial sprays containing 0.02-0.12 % BC had frequently been used in the fitness centres. However, isolates from surfaces upon which antibacterial wipes or antibacterial sprays had not been used were all sensitive to BC. All BC-resistant strains were also resistant to erythromycin, penicillin and ampicillin. In addition, 51 strains showed resistance to cetyltrimethylammonium bromide (CTAB), 15 strains showed resistance to chloramphenicol, 12 strains showed resistance to ciprofloxacin and four strains showed resistance to meticillin. Resistance gene analysis demonstrated that 41 strains contained qacA/B, 30 strains had qacC, 25 strains contained qacG, 16 strains had qacH and eight strains contained qacJ. These data indicate that application of BC is associated with environmental staphylococcal antimicrobial resistance.

  16. The effect of whey isolate and resistance training on strength, body composition, and plasma glutamine.

    Science.gov (United States)

    Cribb, Paul J; Williams, Andrew D; Carey, Michael F; Hayes, Alan

    2006-10-01

    Different dietary proteins affect whole body protein anabolism and accretion and therefore, have the potential to influence results obtained from resistance training. This study examined the effects of supplementation with two proteins, hydrolyzed whey isolate (WI) and casein (C), on strength, body composition, and plasma glutamine levels during a 10 wk, supervised resistance training program. In a double-blind protocol, 13 male, recreational bodybuilders supplemented their normal diet with either WI or C (1.5 gm/kg body wt/d) for the duration of the program. Strength was assessed by 1-RM in three exercises (barbell bench press, squat, and cable pull-down). Body composition was assessed by dual energy X-ray absorptiometry. Plasma glutamine levels were determined by the enzymatic method with spectrophotometric detection. All assessments occurred in the week before and the week following 10 wk of training. Plasma glutamine levels did not change in either supplement group following the intervention. The WI group achieved a significantly greater gain (P < 0.01) in lean mass than the C group (5.0 +/- 0.3 vs. 0.8 +/- 0.4 kg for WI and C, respectively) and a significant (P < 0.05) change in fat mass (-1.5 +/- 0.5 kg) compared to the C group (+0.2 +/- 0.3 kg). The WI group also achieved significantly greater (P < 0.05) improvements in strength compared to the C group in each assessment of strength. When the strength changes were expressed relative to body weight, the WI group still achieved significantly greater (P < 0.05) improvements in strength compared to the C group.

  17. Molecular Characterization of Staphylococcus aureus Isolated from Meat and Their Antibiotic Resistance Profiles

    Directory of Open Access Journals (Sweden)

    Ziad W Jaradat

    2014-01-01

    Full Text Available The aims of this study were to characterize S. aureus isolates from different meat sources in Jordan and study their genetic relationship using PCR-RFLP in addition to their antibiotic resistance profiles. Thirty S. aureus isolates were identified and confirmed by PCR techniques. The isolates from goat and camel meats were sensitive to the majority of the tested antibiotics. Plasmid profiling revealed that 26 isolates contained at least one plasmid with no correlation between the number of plasmids and the resistance profiles. PCR-RFLP of the coagulase gene (coa classified the isolates to several clusters upon digestion with Alu I or Cfo I restriction enzymes. This study concluded that the 30 S. aureus isolates were genetically diverse and comprised heterogeneous population with 7 genotypes at both 33.1 and 51.2 similarity levels.

  18. High level aminoglycoside resistance and distribution of aminoglycoside resistant genes among clinical isolates of Enterococcus species in Chennai, India.

    Science.gov (United States)

    Padmasini, Elango;