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Sample records for negative staphylococci isolated

  1. [Clinical consideration of coagulase negative Staphylococci isolated in blood culture].

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    Oshitani, Yohei; Ishikawa, Tomoyuki; Murata, Ken; Aoyagi, Yoshiki; Yabe, Yasuyo; Aoshima, Masahiro

    2012-01-01

    Despite blood culture's usefulness in antimicrobial therapy, fewer blood cultures and the infrequency of more than 1 set in cultures appear to be problems in Japan. Since June 2007 infection control team (ICT) recommended more than 1 set in blood sampling and intervention in positive blood culture, coagulase negative Staphylococci (CNS) has frequently been isolated from blood culture and its clinical significance is often difficult to judge. To determine the effect of ICT intervention, we evaluated the number of blood culture specimens, the frequency of more than 1 set in all blood culture specimens, and decision-making on antimicrobial treatment for CNS isolated retrospectively from blood. The study was divided into term I in August 2007 to July 2008, term II in August 2008 to July 2009, and term III in August 2009 to February 2010. We also analyzed how physicians treated infection or its suspicion after CNS and its drug susceptibility. The monthly number of blood culture specimens increased from 40.3 to 51.6 between terms I and III. The frequency of more than 1 set in a single blood culture session rose significantly from 67% to 89% between these terms (p blood culture specimens, enable more than 1 set in blood sampling, make it easier to judge the presence of infection, and increase appropriate treatment by physicians. We thus believe that the quality of antimicrobial treatment could be improved through education such as ICT action.

  2. Prevalence and Antimicrobial Susceptibility of Coagulase-Negative Staphylococci isolated from Bovine Mastitis

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    Kaliwal B.B.

    2011-08-01

    Full Text Available The study was carried out to investigate the prevalence and antimicrobial susceptibility of Coagulase-Negative Staphylococci isolated from Bovine Mastitis in and around Dharwad region. A total of 310 samples were screened and 180 confirmed Coagulase-Negative Staphylococci were obtained. The antimicrobial susceptibility of Coagulase-Negative Staphylococci against 10 antimicrobial agents was tested using the disc diffusion method. The highest numbers of Coagulase-Negative Staphylococci were susceptible to ceftriaxone 83.88% followed by cefotaxime 79.41%, methicillin 76.47%, ciprofloxacin 73.52%, erythromycin 70.05%, amikacin 66.11%, gentamycin 42.94%, amoxicillin 36.76%, ampicillin 29.41%, and the lowest susceptibility was shown in penicillin 23.23% . The results indicated that the increase in prevalence and antibiotic resistance pattern of the Coagulase-Negative Staphylococci isolated from bovine mastitis exhibited the highest degree of susceptible to ceftriaxone of all the tested antimicrobial agents. [Vet. World 2011; 4(4.000: 158-161

  3. Coagulase-Negative Staphylococci

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    Heilmann, Christine; Peters, Georg

    2014-01-01

    SUMMARY The definition of the heterogeneous group of coagulase-negative staphylococci (CoNS) is still based on diagnostic procedures that fulfill the clinical need to differentiate between Staphylococcus aureus and those staphylococci classified historically as being less or nonpathogenic. Due to patient- and procedure-related changes, CoNS now represent one of the major nosocomial pathogens, with S. epidermidis and S. haemolyticus being the most significant species. They account substantially for foreign body-related infections and infections in preterm newborns. While S. saprophyticus has been associated with acute urethritis, S. lugdunensis has a unique status, in some aspects resembling S. aureus in causing infectious endocarditis. In addition to CoNS found as food-associated saprophytes, many other CoNS species colonize the skin and mucous membranes of humans and animals and are less frequently involved in clinically manifested infections. This blurred gradation in terms of pathogenicity is reflected by species- and strain-specific virulence factors and the development of different host-defending strategies. Clearly, CoNS possess fewer virulence properties than S. aureus, with a respectively different disease spectrum. In this regard, host susceptibility is much more important. Therapeutically, CoNS are challenging due to the large proportion of methicillin-resistant strains and increasing numbers of isolates with less susceptibility to glycopeptides. PMID:25278577

  4. Changes of Antimicrobial Resistance among Coagulase-Negative Staphylococci Isolated in 8 Consecutive Years in the First Bethune Hospital

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    Xu, Jiancheng; Yan, Chaoying; Wang, Liqiang; Zhou, Qi

    This study was to investigate the antimicrobial resistance of coagulase-negative Staphylococci isolated in 8 consecutive years in the First Bethune Hospital. Disk diffusion test was used to study the antimicrobial resistance. The data were analyzed by WHONET 5 software according to Clinical and Laboratory Standards Institute (CLSI). Most of 2484 strains of coagulase-negative Staphylococci were collected from blood 925 (37.2%), secretions 652 (26.2%) and urine 323 (13.0%) during the past 8 years. The rates of methicillin-resistant coagulase-negative Staphylococci (MRCNS) were between 79.4% and 81.5% during the past 8 years, respectively. In recent 8 years, the antimicrobial resistance of coagulase-negative Staphylococci had increased. Monitoring the antimicrobial resistance to coagulase-negatives Staphylococci should be strengthened. The change of the antimicrobial resistance should be investigated in order to direct rational drug usage in the clinic and prevent bacterial strain of drug resistance from being transmitted.

  5. Antibiotic resistance in coagulase negative staphylococci isolated from wastewater and drinking water.

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    Faria, Cátia; Vaz-Moreira, Ivone; Serapicos, Eduarda; Nunes, Olga C; Manaia, Célia M

    2009-06-01

    This study reports the antibiotic resistance patterns of coagulase negative staphylococci (CNS) isolated from a drinking water treatment plant (WTP), a drinking water distribution network, responsible for supplying water to the consumers (WDN), and a wastewater treatment plant (WWTP), responsible for receiving and treating domestic residual effluents. Genotyping and the 16S rRNA gene sequence analysis demonstrated a higher diversity of species both in the WTP (6 species/19 isolates) and WWTP (12 species/47 isolates) than in the WDN (6 species/172 isolates). Staphylococcus pasteuri and Staphylococcus epidermidis prevailed in the WTP and WDN and Staphylococcus saprophyticus in the WWTP. Staphylococci with reduced susceptibility (resistance or intermediary phenotype) to beta-lactams, tetracycline, clindamycin and erythromycin were observed in all types of water and belonged to the three major species groups. The highest resistance rate was found against erythromycin, presumably due to the presence of the efflux pump encoded by the determinant msrA, detected in the majority of the resistant isolates. This study demonstrates that antibiotic resistant CNS may colonize different types of water, namely drinking water fulfilling all the quality standards.

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

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    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.

  7. Study of antibiotic resistance of staph aureus and coagulase negative staphylococci isolated from patient samples

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    M Anvary

    2006-04-01

    Full Text Available Introduction: Drug resistant Staphylococci are the most important agents of nosocomial infections. In this survey, effect of different antibiotics on these bacteria and their drug resistance was investigated. Methods: The study included 500 strains of Staphylococci. Minimum Inhibitory Concentrations of all antibiotics was determined by the broth macro dilution technique and standard methods from the National Committee for Clinical Laboratory Standards. Result : Oxacillin resistance of S. aureus was 14.2% and that of coagulase-negative staphylococci was 53.4%. The activity of different antibiotics is presented in detail. Conclusion: Surveillance of strains resistant to methicillin is necessary.

  8. Antibiotic Susceptibility of Coagulase-Negative Staphylococci Isolated from Bovine Subclinical Mastitis in Turkey

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    Beytullah Kenar*, Yahya Kuyucuoğlu and Esra Şeker

    2012-06-01

    Full Text Available A total of 572 California Mastitis Test (CMT positive milk samples were collected from 423 lactating cows on 18 private farms in the Middle Western Anatolia. Coagulase–negative staphylococci colonies and CNS species identification was performed based on conventional biochemical techniques and using the API Staph test. Slime production was detected by Congo Red Agar (CRA method. The antibiotic susceptibility was determined according to the National Committee for Clinical Laboratory Standards guidelines (NCCLS. A total of 67 (11.7% coagulase-negative staphylococci (CNS were isolated from CMT positive milk samples. In total, 11 CNS species: S. epidermidis (n=18, S. simulans (n=14, S. warneri (n=10, S. hominis (n=5, S. chromogenes (n=4, S. caprae (n=4, S. xylosus (n=3, S. haemolyticus (n=3, S. hyicus (n=3, S. cohnii (n=2, and S. capitis (n=1 were identified. The most commonly identified CNS species were Staphylococcus epidermidis (26.8% and Staphylococcus simulans (20.8% followed by Staphylococcus warneri (14.9%. Out of 67 CNS isolates, slime production was found in 37 (55.2% CNS strains. CNS isolates were the most resistance to trimethoprim+sulphamethoxazole (76.2%, erythromycin (73.2%, oxacillin and ampicillin (70.2% followed by penicillin (58.3%, gentamicin (53.8%, tetracycline (52.3%, vancomycin (51.8%, ciprofloxacin (26.9%, cefoxitim (23.9%, and cephalothin (13.5%. These results indicate that CNS species are resistant at high rates to the beta-lactam antibiotics which are intensively used in the prevention and treatment of mastitis without any antibiotic susceptibility test in the Middle Western of Turkey.

  9. Enterotoxin genes in coagulase-negative and coagulase-positive staphylococci isolated from bovine milk.

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    de Freitas Guimarães, Felipe; Nóbrega, Diego Borin; Richini-Pereira, Virginia Bodelão; Marson, Pâmela Merlo; de Figueiredo Pantoja, José Carlos; Langoni, Helio

    2013-05-01

    The objective of this study was to isolate and identify the main staphylococcal species causing bovine mastitis in 10 Brazilian dairy herds and study their capability to produce enterotoxins. Herds were selected based on size and use of milking technology, and farms were visited once during the study. All mammary glands of all lactating cows were screened using the California Mastitis Test (CMT) and a strip cup. A single aseptic milk sample (20 mL) was collected from all CMT-positive quarters. Identification of Staphylococcus spp. was performed using conventional microbiology, and PCR was used to determine the presence of enterotoxin-encoding genes (sea, seb, sec, and sed). Of the 1,318 CMT-positive milk samples, Staphylococcus spp. were isolated from 263 (19.9%). Of these isolates, 135 (51%) were coagulase-positive staphylococci (CPS) and 128 (49%) were coagulase-negative staphylococci (CNS). Eighteen different species of CNS were isolated, among which S. warneri, S. epidermidis and S. hyicus were the most frequent. The distribution of Staphylococcus species was different among herds: S. epidermidis was found in 8 herds, S. warneri was found in 7 herds, and S. hyicus in 6 herds. Some of the CNS species (S. saprophyticus ssp. saprophyticus, S. auricularis, S. capitis, and S. chromogenes) were isolated in only one of the farms. Genes related to production of enterotoxins were found in 66% (n=85) of all CNS and in 35% of the CPS isolates. For both CNS and CPS isolates, the most frequently identified enterotoxin genes were sea, seb, and sec; the prevalence of sea differed between CPS (9.5%) and CNS (35.1%) isolates. Staphylococcus warneri isolates showed a greater percentage of sea than seb, sec, or sed, whereas S. hyicus isolates showed a greater percentage of sea than sec. Over 60% of CNS belonged to 3 major species, which carried 62.2 to 81.3% of the enterotoxin genes. The high prevalence highlights the potential for food poisoning caused by these species. For

  10. Coagulase-negative staphylococci strains resistant to oxacillin isolated from neonatal blood cultures

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    Valeria Cataneli Pereira

    2013-11-01

    Full Text Available Coagulase-negative staphylococci (CoNS are the microorganisms most frequently isolated from clinical samples and are commonly found in neonatal blood cultures. Oxacillin is an alternative treatment of choice for CoNS infections; however, resistance to oxacillin can have a substantial impact on healthcare by adversely affecting morbidity and mortality. The objective of this study was to detect and characterise oxacillin-resistant CoNS strains in blood cultures of newborns hospitalised at the neonatal ward of the University Hospital of the Faculty of Medicine of Botucatu. One hundred CoNS strains were isolated and the mecA gene was detected in 69 of the CoNS strains, including 73.2% of Staphylococcus epidermidis strains, 85.7% of Staphylococcus haemolyticus strains, 28.6% of Staphylococcus hominis strains and 50% of Staphylococcus lugdunensis strains. Among these oxacillin-resistant CoNS strains, staphylococcal cassette chromosome mec (SCCmec type I was identified in 24.6%, type II in 4.3%, type III in 56.5% and type IV in 14.5% of the strains. The data revealed an increase in the percentage of CoNS strains isolated from blood cultures from 1991-2009. Furthermore, a predominant SCCmec profile of the oxacillin-resistant CoNS strains isolated from neonatal intensive care units was identified with a prevalence of SCCmec types found in hospital-acquired strains.

  11. Clinical significance of coagulase-negative staphylococci isolates from nosocomial bloodstream infections.

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    Asaad, Ahmed Morad; Ansar Qureshi, Mohamed; Mujeeb Hasan, Syed

    2016-01-01

    Identification of coagulase-negative staphylococci (CoNS) as nosocomial pathogens or contaminants is significant for microbiologists and clinicians. This study aimed to determine the frequency of isolation and antimicrobial resistance patterns of CoNS isolates from nosocomial bloodstream infections (BSIs) and to identify risk factors associated with true bacteremia caused by these emerging pathogens in a Saudi tertiary care hospital. All CoNS-positive cultures from inpatients were identified using the standard methods during a 10-month period. Antimicrobial susceptibility testing was done using the reference broth microdilution method. A total of 208 isolates were identified; of these 75 (32.2%) were considered infection associated, and 133 (67.8%) were considered contamination. S. epidermidis accounted for 34.7% of bacteremia cases, followed by S. hominis (21.3%), S. haemolyticus (16%), and S. saprophyticus (12%). Central venous catheters (p ≤ 0.0001), prior antibiotic therapy (p ≤ 0.0001), the occurrence of more than one positive blood culture (p ≤ 0.0001), and intensive care unit (ICU) admission (p = 0.007) were all independently associated with CoNS bacteremia. Overall, all isolates were highly resistant to penicillin (94.7%), oxacillin (90.7%), and erythromycin (85.3%). The rates of susceptibility to vancomycin, daptomycin, and teicoplanin were 98.7%, 98.7%, and 93.3%, respectively. Our results further highlight that accurate identification and susceptibility testing of CoNS isolates from nosocomial BSIs are crucial to minimize excessive antibiotic use and unnecessary catheter removal. In addition, daptomycin may be an efficient alternative therapeutic option for CoNS resistant to oxacillin and other commonly used antibiotics.

  12. INVESTIGATION OF BIOFILM FORMATION IN COAGULASE-NEGATIVE STAPHYLOCOCCI ISOLATED FROM PLATELET CONCENTRATE BAGS

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    MARTINI, Rosiéli; HÖRNER, Rosmari; RAMPELOTTO, Roberta Filipini; GARZON, Litiérri Razia Litiérri; NUNES, Melise Silveira; TEIXEIRA, Mayza Dalcin; GRAICHEN, Daniel Ângelo Sganzerla

    2016-01-01

    Platelet Concentrates (PCs) are the blood components with the highest rate of bacterial contamination, and coagulase-negative staphylococci (CoNS) are the most frequently isolated contaminants. This study investigated the biofilm formation of 16 contaminated units out of 691 PCs tested by phenotypic and genotypic methods. Adhesion in Borosilicate Tube (ABT) and Congo Red Agar (CRA) tests were used to assess the presence of biofilm. The presence of icaADC genes was assessed by means of the Polymerase Chain Reaction (PCR) technique. With Vitek(r)2, Staphylococcus haemolyticus was considered the most prevalent CoNS (31.25%). The CRA characterized 43.8% as probable biofilm producers, and for the ABT test, 37.5%. The icaADC genes were identified in seven samples by the PCR. The ABT technique showed 85.7% sensitivity and 100% specificity when compared to the reference method (PCR), and presented strong agreement (k = 0.8). This study shows that species identified as PCs contaminants are considered inhabitants of the normal skin flora and they might become important pathogens. The results also lead to the recommendation of ABT use in laboratory routine for detecting biofilm in CoNS contaminants of PCs. PMID:26910444

  13. Contaminating coagulase-negative staphylococci isolated in a lysis-centrifugation (Isolator) blood culture system. Application of different epidemiological markers for deduction of mode of contamination.

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    Arpi, M; Gahrn-Hansen, B; Rosdahl, V T

    1988-07-01

    The lysis-centrifugation blood culture system, Isolator, is a promising system with respect to detection of many significant microorganisms, e.g. Staphylococcus aureus and Enterobacteriaceae, as compared with conventional systems. A drawback of the Isolator system is a disturbingly high rate of clinically insignificant, supposedly contaminating coagulase-negative staphylococci, which leads to considerable waste of time and materials in the laboratory. Several sources of these isolates have been proposed (viz. the patient, the ward environment, the laboratory handling, and the plate media). The aim of this study was to pinpoint the origin of these clinically doubtful coagulase-negative staphylococci, using different epidemiological markers, such as species identification, antibiotic susceptibility patterns, phage-types, and plasmid profiles. Plasmid profile analysis proved to be more discriminating than the other techniques and made it possible to conclude that the laboratory handling of the Isolator system was a major source of coagulase-negative staphylococci in this system.

  14. Frequency of Antiseptic Resistance Among Staphylococcus aureus and Coagulase-Negative Staphylococci Isolated From a University Hospital in Central Iran

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    Taheri, Nona; Ardebili, Abdollah; Amouzandeh-Nobaveh, Alireza; Ghaznavi-Rad, Ehsanollah

    2016-01-01

    Objectives Reduced biocide susceptibility in Staphylococci is associated with various antiseptic resistance genes encoding efflux systems. Our aim was to determine the susceptibility to three disinfectant agents, including benzalkonium chloride (BAC), benzethonium chloride (BZT), and chlorhexidine digluconate (CHDG) among clinical isolates of Staphylococcus aureus and coagulase-negative Staphylococci (CoNS). Methods The minimum inhibitory concentration (MIC) of 60 methicillin-resistant S. aureus (MRSA), 54 methicillin-sensitive S. aureus (MSSA) and 51 CoNS isolates from a single hospital to three biocidal agents (BAC, BZT, and CHDG) was determined. Biocide resistance genes (qacA/B, smr, qacG, qacH, qacJ, and norA) were analyzed by the polymerase chain reaction assay. Results All isolates had MICs for BAC and BZT from 0.25 to 8 µg/mL, and for CHDG from 0.5 to 64 µg/mL. qacA/B was the most common biocide resistance gene among all 165 Staphylococcus isolates (76; 46%), which comprised 38 (63.3%) MRSA, 14 (25.9%) MSSA, and 24 (47%) CoNS. Eleven (6.7%) and 24 (14.5%) isolates among the 165 Staphylococci carried smr and norA genes, respectively. In contrast, other resistance genes such as qacG, qacH, and qacJ were absent in all Staphylococci studied. The qacA/B and smr genes were detected concomitantly in 3% of isolates, and 23.6% strains of the total 165 Staphylococcus isolates were negative for each studied gene. Conclusions The carriage of several biocide resistance genes, including qacA/B, smr, and norA, alone or concurrently, is associated with reduced susceptibility. Use of antiseptics may select for antibiotic-resistant strains and assist their survival in the healthcare environment. PMID:27974958

  15. Antibiotic resistance and mecA characterization of coagulase-negative staphylococci isolated from three hotels in London, UK.

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    Xu, Zhen; Mkrtchyan, Hermine V; Cutler, Ronald R

    2015-01-01

    Antibiotic resistance in bacteria isolated from non-healthcare environments, is a potential problem to public health. In our survey a total of 71 coagulase negative staphylococci (CNS) belonging to 11 different species were isolated from three large hotels in London, UK. The most prevalent species was Staphylococcus haemolyticus, with S. hominis, S. warneri, S. cohnii, and Staphylococcus epidermidis commonly detected. Antimicrobial susceptibilities and carriage of the mecA gene were determined for all of these isolates. Most (85.9%) staphylococci were resistant to multiple antibiotics with all displaying increased susceptibility toward penicillin, fusidic acid, erythromycin, and cefepime. Twenty-one (29.5%) of the isolates were mecA positive, however MIC values to oxacillin, normally associated with the carriage of mecA, varied widely in this group (from 0.06 to 256 mg/L). Fifteen of the twenty-one mecA positive isolates carried SCCmec of these seven were type V, one type I, one type II, and one type IV. Additionally, five of these 15 isolates carried a previously unreported type, 1A, which involves an association between class A mec complex and ccr type 1. The remaining six of the 21 isolates were non-typeable and carried a combination of class A mec complex and ccrC. In addition to this, we also report on new MLST types which were assigned for five S. epidermidis isolates. Four out of these five isolates had MICs between 0.06 and 256 mg/L to oxacillin and would be regarded as clinically susceptible but one isolate had a high oxacillin MIC of 256 mg/L. We demonstrated widespread multiple drug resistance among different staphylococcal species isolated from non-healthcare environments highlighting the potential for these species to act as a reservoir for methicillin and other forms of drug resistance.

  16. Antibiotic resistance and mecA characterization of coagulase-negative staphylococci isolated from three hotels in London, UK

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    Zhen eXu

    2015-09-01

    Full Text Available Antibiotic resistance in bacteria isolated from non–healthcare environments, is a potential problem to public health. In our survey a total of 71 coagulase negative staphylococci belonging to 11 different species were isolated from 3 large hotels in London, UK. The most prevalent species was Staphylococcus haemolyticus, with S. hominis, S. warneri, S.cohnii and S. epidermidis commonly detected. Antimicrobial susceptibilities and carriage of the mecA gene were determined for all of these isolates. Most (85.9% staphylococci were resistant to multiple antibiotics with all displaying increased susceptibility towards penicillin, fusidic acid, erythromycin and cefepime. 21 (29.5% of the isolates were mecA positive, however MIC values to oxacillin, normally associated with the carriage of mecA, varied widely in this group (from 0.06 mg/L to 256 mg/L. 15 of the 21 mecA positive isolates carried SCCmec of these 7 were type V, 1 type I, 1 type II and 1 type IV. Additionally, five of these 15 isolates carried a previously unreported type, 1A, which involves an association between class A mec complex and ccr type 1. The remaining 6 of the 21 isolates were non-typeable and carried a combination of class A mec complex and ccrC. In addition to this, we also report on new MLST types which were assigned for 5 Staphylococcus epidermidis isolates. 4 out of these 5 isolates had MICs between 0.06 to 256 mg/L to oxacillin and would be regarded as clinically susceptible but one isolate had a high oxacillin MIC of 256 mg/L. We demonstrated widespread multiple drug resistance among different staphylococcal species isolated from non-healthcare environments highlighting the potential for these species to act as a reservoir for methicillin and other forms of drug resistance.

  17. Antimicrobial susceptibility of coagulase-negative staphylococci isolated from bovine milk samples

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    Sampimon, O.C.; Lam, T.G.J.M.; Mevius, D.J.; Schukken, Y.H.; Zadoks, R.N.

    2011-01-01

    The aim of this study was to examine whether antimicrobial resistance profiles of coagulase-negative Staphylococcus (CNS) species isolated from milk of dairy cows differed between bacterial species, and to compare results obtained by phenotypic and genotypic profiling of resistance to penicillin, ox

  18. Antimicrobial susceptibility of coagulase-negative staphylococci isolated from bovine milk samples

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    Sampimon, O.C.; Lam, T.G.J.M.; Mevius, D.J.; Schukken, Y.H.; Zadoks, R.N.

    2011-01-01

    The aim of this study was to examine whether antimicrobial resistance profiles of coagulase-negative Staphylococcus (CNS) species isolated from milk of dairy cows differed between bacterial species, and to compare results obtained by phenotypic and genotypic profiling of resistance to penicillin,

  19. Species distribution and antibiotic sensitivity pattern of coagulase negative staphylococci isolated from various clinical specimens

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    Mohan U

    2002-01-01

    Full Text Available A study of 192 strains of Coagulase negative staphylococcus (CONS showed that Staphylococcus epidermidis was the most common species, 158 (82.29% isolated from all clinical specimens followed by S. saprophyticus (30, 15.62% isolated mainly from urine. Slime production was exhibited by 77 (48.7% strains of S. epidermidis and 8 (26.6% of S. saprophyticus and the difference in the slime producing activity was statistically significant (p<0.005. Antibiotic susceptibility testing against 15 commonly used antibiotics showed multidrug resistance with more than 90% resistance to penicillin, more than 50% to cephalexin and ciprofloxacin and more than 20% to methicillin, thus, highlighting the importance of species identification and antibiotic susceptibility testing for clinical isolates of CONS.

  20. Identification of bap and icaA genes involved in biofilm formation in coagulase negative staphylococci isolated from feline conjunctiva.

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    Płoneczka-Janeczko, Katarzyna; Lis, Paweł; Bierowiec, Karolina; Rypuła, Krzysztof; Chorbiński, Paweł

    2014-12-01

    Bap and icaA genes are commonly known to be involved in the biofilm formation. The prevalence of bap and icaA genes and biofilm formation was determined in conjunctival isolates of coagulase negative staphylococci (CNS) collected from cats. The study was conducted on 90 archival CNS isolates collected from feline conjunctiva obtained from clinically healthy cats and cats with ocular problems. Biofilm formation was examined using the microtiter plate (MTP) method. The prevalence of icaA and bap genes was determined using polymerase chain reaction (PCR). Genetic profiles of the bap-positive isolates were examined using the modified random amplified polymorphic DNA (RAPD) method. Of the 90 CNS isolates investigated, 58.9% (53/90) were confirmed to form biofilms on a polystyrene plate after 24 h, and the intensity of the biofilm production varied strongly between positive strains. Among the biofilm-producing isolates, 24.5% (13/53) carried the icaA gene and 3.8% (2/53) carried the bap gene. Among the isolates that did not produce biofilms, the icaA gene and bap gene were detected in 8.1% (3/37) and 2.7% (1/37) of isolates, respectively. This is the first report demonstrating that CNS isolated from feline conjunctiva can potentially be a bap gene reservoir. Preliminary comparison of the genetic profiles of three bap-positive isolates collected from cats showed that each of the isolates has a different genetic background with a high similarity with the human strain of S. epidermidis.

  1. Molecular identification of coagulase-negative staphylococci isolated from bovine mastitis and detection of β-lactam resistance.

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    Srednik, Mariela Elizabeth; Grieben, Mario Andres; Bentancor, Adriana; Gentilini, Elida Raquel

    2015-09-27

    Bovine mastitis is a frequent cause of economic loss in dairy herds. Coagulase-negative staphylococci (CoNS) are increasing in importance as causeof bovine intramammary infection (IMI) throughout the world in recent years. CoNShave been isolated from milk samples collected from cows with clinical andsubclinical mastitis in several countries. Identification of mastitis pathogensis important when selection appropriate antimicrobial therapy. A total of 93 strains of Staphylococcusspp isolated from bovine clinical andsubclinical mastitis in Argentina during 2010-2013 were identified by Restriction Fragment Length Polymorphism (RFLP)-PCR using the gap gene. The isolates were tested by PCR for the presence of blaZ gene and mecA gene and were tested by disk diffusion for the susceptibilityto penicillin and cefoxitin. The most common CoNS species was S.chromogenes 46.2% (43/93), followed by S. devriesei 11.8% (11/93) and S. haemolyticus 9.7% (9/93). The blaZ gene was detected in 19 (20.4%) but only 16 (17.2%) isolates were resistant to penicillin; the mecA was detected in6 (6.5%) isolates but only 4 (4.3) were resistant to cefoxitin. The 6 mecA-positive isolates showed oxaxillinMICs ≤ 0.5 μg/ml. CoNSare important minor mastitis pathogens and can be the cause of substantial economic losses. Despite the low resistance to PEN in Argentina, the presenceof MR isolates found in this study emphasize the importance of identificationof CoNS when an IMI is present because of the potentially risk of lateraltransfer of resistance genes between staphylococcal species.

  2. Polymerase chain reaction detection of enterotoxins genes in coagulase-negative staphylococci isolated from Brazilian Minas cheese.

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    Rall, Vera Lúcia Mores; Sforcin, José Maurício; de Deus, Maria Fernanda Ramos; de Sousa, Daniel Casaes; Camargo, Carlos Henrique; Godinho, Natália Cristina; Galindo, Luciane Almeida; Soares, Taíssa Cook Siqueira; Araújo, João Pessoa

    2010-09-01

    For a long time, Staphylococcus aureus has been always thought to be the only pathogenic species among Staphylococcus, while coagulase-negative staphylococci (CNS) were classified as contaminant agents. However, molecular techniques have shown that these microorganisms also possess enterotoxin-encoding genes. The aim of this study was to analyze the frequency of genes for staphylococcal enterotoxins SEA, SEB, SEC, and SED in CNS strains isolated from Minas soft cheese and to assess the in vitro production of toxins. CNS were found in 65 (72.2%) samples of cheese: 23 were Staphylococcus saprophyticus, 16 Staphylococcus warneri, 10 Staphylococcus epidermidis, 9 Staphylococcus xylosus, 3 Staphylococcus haemolyticus, 2 Staphylococcus schleiferi subsp. schleiferi, and 1 each Staphylococcus capitis subsp. urealyticus and Staphylococcus caprae. Seventeen (26.2%) CNS strains had genes for enterotoxins, and sea was more frequently found (18.5%), followed by sec in three and seb in two strains, whereas the sed gene was not found. S. saprophyticus showed enterotoxin genes in 6 of 23 isolates, but only sea was observed. On the other hand, five strains of S. warneri showed the sea, seb, or sec gene. In spite of the presence of these enterotoxin genes, these strains did not produce enterotoxins in vitro. It is essential to understand the real role of CNS in food, and based on the presence of enterotoxin genes, CNS should not be ignored in epidemiological investigations of foodborne outbreaks.

  3. 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

  4. Prevalence and antibiotic resistance of coagulase-negative Staphylococci isolated from poultry farms in three regions of Ghana

    DEFF Research Database (Denmark)

    Boamah, Vivian Etsiapa; Agyare, Christian; Odoi, Hayford

    2017-01-01

    The use of antibiotics in animal production has been associated with the development and spread of antibiotic-resistant organisms including commensals. Coagulase-negative Staphylococcus (CoNS) species, which were until recently considered non-pathogenic, have been associated with opportunistic...... infections and high resistance to several antibiotics. This study sought to determine the prevalence, identity, and phenotypic resistance of coagulase-negative Staphylococcus spp. isolated from some selected poultry farms and farm workers in the Ashanti, Brong Ahafo, and Greater Accra regions of Ghana...... (57.03%), doxycycline (43.75%), and oxacillin (43.36%). Multi-drug resistance (MDR) was observed in 19.14% of the isolates. MDR was higher in isolates obtained from poultry farm workers (61.22%) than isolates from poultry litter (38.78%). The above findings call for stricter monitoring of antibiotic...

  5. 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.

  6. Prevalence and antibiotic resistance of coagulase-negative Staphylococci isolated from poultry farms in three regions of Ghana

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    Boamah VE

    2017-06-01

    Full Text Available Vivian Etsiapa Boamah,1 Christian Agyare,1 Hayford Odoi,1 Francis Adu,1 Stephen Yao Gbedema,1 Anders Dalsgaard2 1Microbiology Section, Department of Pharmaceutics, Faculty of Pharmacy and Pharmaceutical Sciences, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana; 2Section of Food Safety and Zoonoses, Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederisksberg, Denmark Abstract: The use of antibiotics in animal production has been associated with the development and spread of antibiotic-resistant organisms including commensals. Coagulase-negative Staphylococcus (CoNS species, which were until recently considered non-pathogenic, have been associated with opportunistic infections and high resistance to several antibiotics. This study sought to determine the prevalence, identity, and phenotypic resistance of coagulase-negative Staphylococcus spp. isolated from some selected poultry farms and farm workers in the Ashanti, Brong Ahafo, and Greater Accra regions of Ghana. Poultry litter samples and oral swabs of poultry farm workers were collected, from which bacterial species were isolated, identified, and analyzed. Various selective media were used for the presumptive identification of the different species. Confirmation of bacterial identity was done using matrix-assisted laser desorption/ionization time of flight (MALDI-TOF mass spectrometry. Antibiotic susceptibility testing of the isolates was performed using the Kirby-Bauer disk diffusion method. Zones of growth inhibition were interpreted based on the European Committee on Antimicrobial Susceptibility Testing (EUCAST guidelines. Two hundred and fifty-six coagulase-negative Staphylococcus spp., comprising S. sciuri (42.97%, S. lentus (35.94%, S. gallinarum (6.64%, S. xylosus (4.30%, S. haemolyticus (3.91%, S. saprophyticus (1.95%, and S. cohnii (0.39% were confirmed by MALDI-TOF. CoNS were isolated from samples

  7. Production of antilisterial bacteriocins by staphylococci isolated from bovine milk

    Science.gov (United States)

    A collection of 111 staphylococcal isolates recovered from healthy cows in 41 dairy herds in Brazil was surveyed for the production of bacteriocins. The group included 94 coagulase positive and 17 coagulase negative strains of staphylococci. All cultures were grown in tryptic soy broth for 18 h at ...

  8. Prevalence of coagulase-negative staphylococci and determination of antimicrobial resistance in accompany with types of SCCmec in isolated of nosocomial infections

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    Mohammad Reza Arabestani

    2016-03-01

    Full Text Available Background: Coagulase-negative staphylococci (CoNS were considered as contaminats previously, but, during the past decade considered as one of the most common photogenic bacteria in hospital. Resistance to beta-lactams especially methicillin in staphylococcus species is being worrying in hospitals. Rapid identification of mechanisms of resistance and confirmation of their resistance to methicillin is a basic principle for antibiotic treatment. The aim of this study was to determine antibiotic resistance, frequency of mecA gene, and determination of SCCmec types in CoNS isolates from teaching hospitals in Iran. Methods: The descriptive cross-sectional study was carried out one hundred clinical samples isolated from patients with an average age of 7-69 years at teaching hospitals in Hamadan City, Iran, from September 2014 to February 2015. After confirmation of isolates by microbiological standard biochemical tests, antimicrobial susceptibility testing was performed by disk agar diffusion (DAD method. After extraction of isolated genomicm, mecA gene was detected. Then, the types of SCCmec were performed by PCR. Results: In this study, 387 clinical samples were collected which among 100 CoNS isolated, Staphylococcus epidermidis was the most prevalent species with frequency 55 (55%, followed by S. haemolyticus 40(40% and S. saprophyticus 5(5%. The highest antibiotic susceptibility was to rifampin 96(96% and the lowest resistance was detected for trimethoprim/sulfamethoxazole (TMP/SMX 47(47%. None of the strains were resistant to vancomycin. Resistance to methicillin was detected in 50% of CoNS isolates. Typing of SCCmec was performed by The polymerase chain reaction (PCR. Frequency types of SCCmec was type III with frequency 13(13%, type V 11(11%, type II 6(6%, type IV 4 (4%, type I 3(3% respectively. Thirteen isolated was not typable in this study. Conclusion: The result of this study showed that a large percentage of coagulase-negative

  9. Studies on prevalence of biofilm associated genes and primary observation on sasX gene in clinical isolates of coagulase negative staphylococci (CoNS).

    Science.gov (United States)

    Soumya, Kandammuriyil Radhakrishnan; Mathew, Shiji; Sugathan, Sheela; Mathew, Jyothis; Radhakrishnan, Edayileveettil Krishnankutty

    2016-04-01

    Coagulase negative staphylococci (CoNS) are nosocomial pathogens that cause indwelling medical device associated infections due to its biofilm forming potential and multiple antibiotic resistance. The current study focused on species identification, antibiotic resistance profile and molecular basis of biofilm formation and attachment of CoNS isolated from clinical samples. Along with this, molecular screening for mecA and newly identified surface colonization protein encoded by sasX gene was also conducted. S. epidermidis (n = 19, 47%) was identified as the most prevalent CoNS species and very interestingly two biofilm forming, mecA positive S. epidermidis isolates were found to carry all the biofilm associated genes screened in this study, which indicates its potential to form the strong biofilm. Another novel observation of the study is the detection of sasX gene in one biofilm positive S. epidermidis isolate. The study also identified one doxycycline resistant mecA positive, multidrug resistant S. haemolyticus isolate. In conclusion, the study signifies the existence of multiple biofilm related genes, multidrug resistance and the presence of sasX gene among clinical isolates of CoNS.

  10. Prosthetic Joint Infections Due to Coagulase-Negative Staphylococci

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    Mohamad

    2015-12-01

    Full Text Available Context Staphylococcus aureus and coagulase-negative staphylococci (CoNS are the most frequently isolated pathogens and are responsible for approximately two-thirds of joint replacement infections. Evidence Acquisition Although both belong to the staphylococci genus, there are several epidemiological and therapeutic differences between S. aureus and the heterogenous group of CoNS. In general, however, preventive and therapeutic recommendations for Prosthetic Joint Infection PJI due to CoNS do not differ from PJI caused by other pathogens. Results The main differences between the pathogens lie in the clinical presentation of PJI, the presumed origin of infection, and the presence of a higher proportion of methicillin-resistant strains leading to a limited choice of antibiotic agents. Conclusions Fortunately, due to its lower virulence as compared to its cousin S. aureus, PJI due to CoNS may display higher remission rates than S. aureus-caused PJI after combined surgical and medical management.

  11. Identification of Staphylococci Isolated from Chickens and the Determination of Their Susceptibility to Some Antibiotics

    OpenAIRE

    ALTAY, Gülay

    2014-01-01

    In this study, a total of 120 staphylococci strains, 46 coagulase positive and 74 coagulase negative, isolated from chickens were identified according to the characteristics of their cultural, biochemical and some antibiotic properties. The coagulase positive staphylococci consisted of 28 S. aureus, 9 S. delphini, 3 S. intermedius, 2 S. aureus subsp. anaerobicus, 1 S. schleiferi subsp. coagulans, 1 S. hyicus, and 2 unidentified strains. The coagulase negative staphylococci were identified as...

  12. Prevalence, antimicrobial resistance, and virulence characteristics of mecA-encoding coagulase-negative staphylococci isolated from soft cheese in Brazil.

    Science.gov (United States)

    Fontes, Cláudia Oliveira; Silva, Vânia Lúcia; de Paiva, Mayara Rodrigues Brandão; Garcia, Rafaela Alvim; Resende, Juliana Alves; Ferreira-Machado, Alessandra Barbosa; Diniz, Cláudio Galuppo

    2013-04-01

    Coagulase-negative staphylococci (CoNS), which are generally neglected as foodborne bacteria, are emerging as significant opportunistic pathogens that may be highly resistant to available antimicrobial drugs. In this study, antimicrobial susceptibility patterns, mecA gene occurrence, and virulence-associated characteristics were evaluated in CoNS isolated from soft cheese in Brazil. A total of 227 bacterial isolates were recovered from 35 cheese samples belonging to 5 batches with 7 different trademarks. The CoNS counts ranged from 10(6) to 10(7) CFU/g. High antimicrobial resistance percentages were observed for oxacillin (76.2%), penicillin (78.5%), erythromycin (67.8%), gentamicin (47.2%), clindamycin (35.7%), rifampicin (26.8%), azithromycin (14.7%), tetracycline (14.7%), levofloxacin (14.2%), and sulfamethoxazole-trimethoprim (11.9%). A low antimicrobial resistance percentage was observed for chloramphenicol (2.3%), and all of the tested bacteria were susceptible to vancomycin and linezolid. In total, a multiple antibiotic resistance (MAR) index of >0.2 was observed for 80.6% of the isolated CoNS. However, the MAR index ranged from 50% to 92.6% when only bacterial cheese isolates belonging to the same trademark were considered. Regarding to the prevalence of CoNS carrying mecA gene, 81.5% of the isolated strains were mecA(+) , and 76.2% of these were phenotypically resistant to oxacillin. Three isolates carried the enterotoxin A gene (sea), 29.5% produced biofilm in a laboratory test, and α- or ß-hemolysis were observed for 3% and 5.2%, respectively. This study highlights the extent of the antimicrobial resistance phenomenon in neglected foodborne microorganisms and the potential public health risks that are related to the consumption of CoNS-contaminated soft cheese.

  13. Short communication: Prevalence of methicillin resistance in coagulase-negative staphylococci and Staphylococcus aureus isolated from bulk milk on organic and conventional dairy farms in the United States.

    Science.gov (United States)

    Cicconi-Hogan, K M; Belomestnykh, N; Gamroth, M; Ruegg, P L; Tikofsky, L; Schukken, Y H

    2014-05-01

    The objective of this study was to evaluate the presence of methicillin-resistant Staphylococcus aureus and coagulase-negative Staphylococcus spp. in bulk tank milk samples from 288 organic and conventional dairy farms located in New York, Wisconsin, and Oregon from March 2009 to May 2011. Due to recent publications reporting the presence mecC (a mecA homolog not detected by traditional mecA-based PCR methods), a combination of genotypic and phenotypic approaches was used to enhance the recovery of methicillin-resistant organisms from bulk tank milk. In total, 13 isolates were identified as methicillin resistant: Staph. aureus (n=1), Staphylococcus sciuri (n=5), Staphylococcus chromogenes (n=2), Staphylococcus saprophyticus (n=3), Staphylococcus agnetis (n=1), and Macrococcus caseolyticus (n=1). The single methicillin-resistant Staph. aureus isolate was identified from an organic farm in New York, for an observed 0.3% prevalence at the farm level. The methicillin-resistant coagulase-negative staphylococci prevalence was 2% in the organic population and 5% in the conventional population. We did not identify mecC in any of the isolates from our population. Of interest was the relatively high number of methicillin-resistant Staph. sciuri recovered, as the number of isolates from our study was considerably higher than those recovered from other recent studies that also assessed milk samples. Our research suggests that the presence of a potential methicillin-resistant Staphylococcus reservoir in milk, and likely the dairy farm population in the United States, is independent of the organic or conventional production system.

  14. Prevalence of antibiotic resistance in multi-drug resistant coagulase-negative staphylococci isolated from invasive infection in very low birth weight neonates in two Polish NICUs.

    Science.gov (United States)

    Brzychczy-Wloch, Monika; Borszewska-Kornacka, Maria; Gulczynska, Ewa; Wojkowska-Mach, Jadwiga; Sulik, Malgorzata; Grzebyk, Monika; Luchter, Malgorzata; Heczko, Piotr B; Bulanda, Malgorzata

    2013-12-20

    Multi-drug resistant coagulaso-negative staphylococci (CNS) have become an increasing problem in nosocomial infections connected with the presence of medical devices. The paper aimed to analyze the prevalence of antibiotic resistance in CNS isolated from invasive infection in very low birth weight (VLBW) neonates. Continuous prospective target surveillance of infections was conducted in 2009 at two Polish NICUs that participated in the Polish Neonatology Surveillance Network (PNSN). The study covered 386 neonates with VLBW (≤1500 g), among which 262 cases of invasive infection were detected with predominance of CNS (123; 47%). Altogether, 100 CNS strains were analyzed. The resistance phenotypes were determined according to EUCAST. Resistance genes: mecA, ermA, ermB, ermC, msrA, aac(6')/aph(2''), ant(4')-Ia and aph(3')-IIIa were detected using multiplex PCR. The most common species was S. epidermidis (63%), then S. haemolyticus (28%) and other CNS (9%). Among S. epidermidis, 98% of isolates were resistant to methicillin, 90% to erythromycin, 39% to clindamycin, 95% to gentamicin, 60% to amikacin, 36% to ofloxacin, 2% to tigecycline, 3% to linezolid and 13% to teicoplanin. Among S. haemolyticus isolates, 100% were resistant to methicillin, erythromycin and gentamicin, 18% to clindamycin, 50% to amikacin, 86% to ofloxacin, 14% to tigecycline and 4% to teicoplanin. No resistance to linezolid was detected for S. haemolyticus isolates. Moreover, all isolates of S. epidermidis and S. haemolyticus were susceptible to vancomycin. The mecA gene was detected in 98% of S. epidermidis isolates and all of S. haemolyticus ones. Among macrolide resistance isolates, the ermC was most common in S. epidermidis (60%) while msrA was prevalent in S. haemolyticus (93%). The ermC gene was indicated in all isolates with cMLSB, whereas mrsA was found in isolates with MSB phenotype. Of the aminoglycoside resistance genes, aac(6')/aph(2'') were present alone in 83% of S. epidermidis

  15. Licheniocin 50.2 and Bacteriocins from Lactococcus lactis subsp. lactis biovar. diacetylactis BGBU1-4 Inhibit Biofilms of Coagulase Negative Staphylococci and Listeria monocytogenes Clinical Isolates

    Science.gov (United States)

    Draganic, Veselin; Lozo, Jelena; Beric, Tanja; Kojic, Milan; Arsic, Biljana; Garalejic, Eliana; Djukic, Slobodanka; Stankovic, Slavisa

    2016-01-01

    Background Coagulase negative staphylococci (CoNS) and Listeria monocytogenes have important roles in pathogenesis of various genital tract infections and fatal foetomaternal infections, respectively. The aim of our study was to investigate the inhibitory effects of two novel bacteriocins on biofilms of CoNS and L. monocytogenes genital isolates. Methods The effects of licheniocin 50.2 from Bacillus licheniformis VPS50.2 and crude extract of bacteriocins produced by Lactococcus lactis subsp. lactis biovar. diacetylactis BGBU1-4 (BGBU1-4 crude extract) were evaluated on biofilm formation and formed biofilms of eight CoNS (four S. epidermidis, two S. hominis, one S. lugdunensis and one S. haemolyticus) and 12 L. monocytogenes genital isolates. Results Licheniocin 50.2 and BGBU1-4 crude extract inhibited the growth of both CoNS and L. monocytogenes isolates, with MIC values in the range between 200–400 AU/ml for licheniocin 50.2 and 400–3200 AU/ml for BGBU1-4 crude extract. Subinhibitory concentrations (1/2 × and 1/4 × MIC) of licheniocin 50.2 inhibited biofilm formation by all CoNS isolates (p bacteriocins in concentrations of 100 AU/mL and 200 AU/mL reduced the amount of 24 h old CoNS and L. monocytogenes biofilms (p bacteriocins have potential to be used for genital application, to prevent biofilm formation and/or to eradicate formed biofilms, and consequently reduce genital and neonatal infections by CoNS and L. monocytogenes. PMID:27930711

  16. Prevalence and antimicrobial susceptibility pattern of coagulase-negative staphylococci (CoNS isolated from clinical specimens in Northern of Jordan

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    Ibrahim Ali Al Tayyar

    2016-01-01

    Full Text Available Background: Coagulase negative Staphylococci (CoNS are one of the most common bacteria found on human skin and on mucous membranes as a component of normal flora. The presence of CoNS in clinical specimens is frequently associated with an infectious aetiology or contamination.Objectives: We aimed to evaluate CoNS species distribution and susceptibility patterns in specimens obtained from clinics and hospitals in the Northern area of Jordan.Methods: Standard identification methods showed the presence of CoNS in 223 specimens at different local hospitals. Susceptibility testing was performed using 18 antibiotics in accordance with the Clinical and Laboratory Standards Institute (CLSI recommendations.Results: Staphylococcus epidermidis and S. haemolyticus were found to be the most common species isolated from all spec- imens representing 122 (54.7% and 52 (23.4% of all CoNS species, respectively. Antibiotic susceptibility testing of CoNS species revealed their sensitivity to vancomycin, linozolid, rifampin and nitrofurantin, while showing a highly resistant pattern to ampicillin, penicillin, ceftriaxone, cefazolin, amoxicillin-clavulanic acid and erythromycin. Some variation of the susceptibility pattern of CoNS species were identified in specimens isolated from the ICU and paediatric hospital wards as well as from clinical specimens of urine, blood and catheter tips.Conclusion: The most common CoNS isolates were found to be S. epidermidis and S. haemolyticus with variable percentag- es according to the specimen source. Moreover, a high susceptibility CoNS to vancomycin, rifampin, and linezolid showed resistance to amoxicillin and penicillin. Keywords: CoNS, Antibiotic, Hospital, Intensive Care Units, Hospital Wards, Nosocomial Infection

  17. Prevalence of coagulase-negative staphylococci in bovine mastitis in Zimbabwe

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    T. Kudinha

    2002-07-01

    Full Text Available This study was carried out to determine the prevalence of coagulase-negative staphylococci in clinical and subclinical mastitis in commercial and small-scale farms in Zimbabwe. Thirty five quarter milk samples from clinical mastitis cases and 371 quarter milk samples from cows with subclinical mastitis were cultured for bacterial pathogens. The most frequent pathogens isolated in clinical mastitis were the enteric bacteria (31.4 %, followed by coagulase negative staphylococci (22.9 % and then Staphylococcus aureus (17.1 %, whereas in subclinical mastitis S. aureus (34.2 % and coagulase-negative staphylococci were (33.2 % the most common. Bacillus species were only isolated in milk samples from subclinical mastitis. Coagulase-negative staphylococci were observed in mixed infections with other bacteria in only 2.2 % of the 406 milk samples from clinical and subclinical mastitis where they were isolated together with Bacillus species in 6 of the 9 mixed infection cases. About 95 % of the milk samples from which 131 coagulase-negative staphylococci were isolated had correspondingly high somatic cell counts. The coagulase-negative staphylococci isolated most frequently were S. chromogenes (7.9 %, S. epidermidis (7.4 % and S. hominis (5.9 %. They were all associated with high somatic cell counts. All the coagulase-negative staphylococci isolates were susceptible to cloxacillin and erythromycin, and more than 90 %of the isolates were susceptible to neomycin, penicillin and streptomycin. The highest resistance was to tetracycline (17.6 %, followed by lincomycin (13.7 %. About 8 % of the isolates were resistant to both penicillin and streptomycin.

  18. Antimicrobial resistance of coagulase-negative staphylococci and lactic acid bacteria from industrially produced dairy products

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    Nevijo Zdolec

    2013-03-01

    Full Text Available In this research, the susceptibility to clindamycin, tetracycline, amikacin, amoxicillin + clavulanic acid, enrofloxacine, vancomycin, trimethoprim + sulphametoxazol, tobramycin, chloramphenicol, ciprofloxacin, erythromycin, penicillin and trimethoprim was tested in coagulase-negative staphylococci (n=78 and lactic acid bacteria (n=30 by means of disk diffusion test and E-test. The isolates were collected from soft and hard cheeses, butter and brine. All isolates of coagulase-negative staphylococci were susceptible to clindamycin, amikacin, amoxicillin + clavulanic acid, enrofloxacine, vancomycin, chloramphenicol and ciprofloxacin according to CLSI breakpoints. A total of 30 staphylococci isolates (38.46 % were resistant to erythromycin, 18 to penicillin (23.07 %, 4 to tetracycline (5.12 %, and one isolate to trimethoprim, tobramicin and trimethoprim + sulphametoxazol (1.28 %. Among 78 tested staphylococci, 35 of them were resistant to at least one antimicrobial substance (44.87 %. The rate of resistant isolates of different soft cheese types ranged from 22 to 70 %, while resistant staphylococci were absent in hard cheese and brine. The growth of lactic acid bacteria was not influenced by trimethoprim + sulphametoxazol (n=29, vancomycin (n=29, trimethoprim (n=28, amikacin (n=10 and tobramycin (n=10. The results show that significant part of apathogenic microbiota in different dairy products is phenotypically resistant to antimicrobial agents.

  19. Antimicrobial susceptibility of coagulase-negative Staphylococci isolated from mastitic cattle in Brazil Susceptibilidade antimicrobiana de cepas de Staphylococci coagulase-negativa isoladas de leite de bovinos com mastite no Brasil

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    T.R.O. Machado

    2008-02-01

    Full Text Available Um total de 109 cepas de Staphylococci coagulase-negativa foi isolado de leite de vacas com mastite clínica e subclínica, em 35 fazendas, situadas em nove estados brasileiros, no período de fevereiro a maio de 2005. Os isolados foram investigados em relação a susceptibilidade in vitro a diversos agentes antimicrobianos. A resistência à penicilina foi a observação mais freqüente (93,5%, seguida por sulfonamida (88,9%, novobiocina (88,6% e ampicilina (85,3%. Todas as cepas examinadas mostraram resistência a pelo menos uma das drogas antimicrobianas testadas. Cepas apresentando resistência múltipla foram extremamente comuns, com 10,0% dos microrganismos isolados apresentando resistência a todas as drogas antimicrobianas. Os resultados obtidos indicaram que as cepas de Staphylococci coagulase-negativas, isoladas no Brasil, apresentaram um alto grau de resistência a antimicrobianos. Estes resultados são, provavelmente, uma conseqüência da pressão devida ao uso intensivo de drogas antimicrobianas.

  20. Antimicrobial activities of ozenoxacin against isolates of propionibacteria and staphylococci from Japanese patients with acne vulgaris.

    Science.gov (United States)

    Nakajima, Akiko; Ikeda, Fumiaki; Kanayama, Shoji; Okamoto, Kazuaki; Matsumoto, Tatsumi; Ishii, Ritsuko; Fujikawa, Akira; Takei, Katsuaki; Kawashima, Makoto

    2016-08-01

    Ozenoxacin, a novel non-fluorinated topical quinolone, was assessed for in vitro antimicrobial activity against clinical isolates of propionibacteria and staphylococci according to the broth microdilution method recommended by the Clinical and Laboratory Standards Institute. The isolates used in this study were collected from Japanese patients with acne vulgaris during a period from 2012 to 2013. The MIC90s of ozenoxacin against Propionibacterium acnes (n=266), Propionibacterium granulosum (n=10), Staphylococcus aureus (n=23), Staphylococcus epidermidis (n=229) and other coagulase-negative staphylococci (n=82) were ≤0.06, ≤0.06, ≤0.06, 0.125 and ≤0.06 µg ml-1, respectively. The antimicrobial activity of ozenoxacin against the clinical isolates of propionibacteria and staphylococci was greater than that of five reference antimicrobial agents which have been used for the treatment of acne vulgaris. The MICs of ozenoxacin were correlated with those of nadifloxacin in P. acnes and S. epidermidis isolates. However, the MICs of ozenoxacin were 0.25-0.5 µg ml-1 and 0.5-8 µg ml-1 against nadifloxacin-resistant P. acnes (MIC: ≥8 µg ml-1; n=8) and S. epidermidis (MIC: ≥64 µg ml-1; n=10), respectively. These results indicated the potent antimicrobial activity against P. acnes and S. epidermidis isolates resistant to nadifloxacin. Topical ozenoxacin could represent an alternative therapeutic drug for acne vulgaris based on its potent antimicrobial activity against the isolates of propionibacteria and staphylococci from acne patients.

  1. Methicillin-resistant coagulase-negative staphylococci from healthy dogs in Nsukka, Nigeria

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    Kennedy F. Chah

    2014-01-01

    Full Text Available The occurrence, resistance phenotype and molecular mechanisms of resistance of methicillin-resistant staphylococci from groin swabs of 109 clinically healthy dogs in Nsukka, Nigeria were investigated. The groin swab samples were cultured on mannitol salt agar supplemented with 10 µgof cloxacillin. Sixteen methicillin-resistant coagulase negative staphylococci (MRCoNS, all harbouring the mecA gene were isolated from 14 (12.8% of the 109 dogs studied. The MRCoNS isolated were: S. sciuri subspecies rodentium, S. lentus, S. haemolyticus, and S. simulans with S. sciuri subspecies rodentium (62.5% being the predominant species. Thirteen (81.3% of the MRCoNS were resistant to tetracycline while 12 (75% and 10 (62.5% were resistant to kanamycin and trimthoprim-sulphamethoxazole respectively. None of the isolates was resistant to fusidic acid, linezolid and vancomycin. Thirteen (81.3% of the MRCoNS were multi-drug resistance (MDR. Other antimicrobial genes detected were: blaZ, tet(K, tet(M, tet(L, erm(B, lnu(A, aacA-aphD, aphA3, str, dfr(G, cat pC221,and cat pC223. Methicillin-resistant staphylococci are common colonizers of healthy dogs in Nigeria with a major species detected being S. sciuri subsp. rodentium.

  2. Isolation of methicillin-resistant staphylococci in the dental operatory.

    Science.gov (United States)

    Horiba, N; Yoshida, T; Suzuki, K; Maekawa, Y; Ito, M; Matsumoto, T; Nakamura, H

    1995-01-01

    The state of contamination by methicillin-resistant Staphylococci, including methicillin-resistant Staphylococcus aureus (MRSA) at the conservative dentistry section of the School of Dentistry, Aichi-Gakuin University, was investigated. Methicillin-resistant coagulase-negative Staphylococci (MRCNS) were detected in 8 (20.5%) of 39 dental staff (28 dentists, 8 dental hygienists, and 3 dental assistants), but MRSA was not detected. MRCNS was also detected in locations such as the dental chair, dental bracket, dental cabinet, and floor, as well as in airborne samples. The presence of the same strain in both the oral cavity of staff persons and the dental chair/floor locations was determined by biochemical properties, antibiotic sensitivity tests, and electrophoretic patterns.

  3. Nasal Carriage of Uncommon Coagulase-Negative Staphylococci in Nurses and Physicians of Tehran University Hospitals

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    Elaheh Salimi

    2016-05-01

    Full Text Available Coagulase-negative staphylococci (CoNS have been identified as a major cause of nosocomial infections. Nasal carriage of CoNS in nurses and physicians is known to be an important risk factor for potential hospital infections. This study was carried out to investigate the prevalence of nasal carriage of uncommon coagulase-negative staphylococci among nurse and physician staffs of Tehran University Hospitals. A total of 116 CoNS were isolated from anterior nares of the study participants working in different wards of the hospitals. Thirteen uncommon CoNS were identified using phenotypic and biochemical methods, were subsequently confirmed by API kits. Staphylococcus xylosus, Staphylococcus haemolyticus, and Staphylococcus capitis species accounted for 53.85%, 30.77%, and 15.38% from the isolates, respectively. Six isolates (46.15% were found to be resistant to methicillin. In conclusion, screening of healthcare workers for uncommon CoNS colonization along with identification and testing for susceptibility of cultured isolates is of paramount importance in strengthening effective nosocomial infection control and prevention measures.

  4. Development and evaluation of hexaplex PCR for rapid detection of methicillin, cadmium/zinc and antiseptic-resistant staphylococci, with simultaneous identification of PVL-positive and -negative Staphylococcus aureus and coagulase negative staphylococci.

    Science.gov (United States)

    Panda, Sasmita; Kar, Sarita; Choudhury, Ranginee; Sharma, Savitri; Singh, Durg V

    2014-03-01

    We developed a multiplex PCR to detect the presence of methicillin- (mecA), cadmium/zinc-(czrC) and antiseptic-resistant (qacA/B) staphylococci and to identify Panton-Valentine leukocidin (PVL)-positive and -negative Staphylococcus aureus and coagulase-negative staphylococci (CoNS) from infected and healthy eyes. The assay was validated on 177 staphylococci comprising of 55 each of S. aureus and CoNS isolated from infected eyes and five S. aureus and 62 CoNS isolated from healthy eyes and nine direct ocular samples. Nine direct ocular samples for in situ testing consisted of corneal scrapings (4), conjunctiva swabs (2) and others (3). Multiplex PCR result was correlated with genotype data obtained with single PCR and dot-blot assay. The control strains that were positive in multiplex PCR for 16S rRNA, nuc, mecA, pvl, czrC and qacA/B genes were also positive in the dot-blot assay. The specificity of amplified genes obtained with reference strains was further confirmed by DNA sequencing. The single step-hexaplex PCR method can be used for rapid detection of mecA, nuc, pvl, czrC and qacA/B genes in staphylococci with simultaneous identification of PVL-positive and -negative S. aureus and CoNS from a variety of ocular samples.

  5. Glucose & sodium chloride induced biofilm production & ica operon in clinical isolates of staphylococci

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    Astha Agarwal

    2013-01-01

    Full Text Available Background & objectives: All colonizing and invasive staphylococcal isolates may not produce biofilm but may turn biofilm producers in certain situations due to change in environmental factors. This study was done to test the hypothesis that non biofilm producing clinical staphylococci isolates turn biofilm producers in presence of sodium chloride (isotonic and high concentration of glucose, irrespective of presence or absence of ica operon. Methods: Clinical isolates of 100 invasive, 50 colonizing and 50 commensal staphylococci were tested for biofilm production by microtiter plate method in different culture media (trypticase soy broth alone or supplemented with 0.9% NaCl/ 5 or 10% glucose. All isolates were tested for the presence of ica ADBC genes by PCR. Results: Biofilm production significantly increased in the presence of glucose and saline, most, when both glucose and saline were used together. All the ica positive staphylococcal isolates and some ica negative isolates turned biofilm producer in at least one of the tested culture conditions. Those remained biofilm negative in different culture conditions were all ica negative. Interpretation & conclusions: The present results showed that the use of glucose or NaCl or combination of both enhanced biofilm producing capacity of staphylococcal isolates irrespective of presence or absence of ica operon.

  6. In vitro additive effect of imipenem combined with vancomycin against multiple-drug resistant, coagulase-negative Staphylococci.

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    Traub, W H; Spohr, M; Bauer, D

    1986-09-01

    Imipenem combined with vancomycin resulted in a marked additive effect in vitro against 9 clinical isolates of multiple-drug resistant (MDR), coagulase-negative staphylococci, including strains resistant against imipenem. The additive effect was documented with the aid of checkerboard MIC determinations and with time kill curve experiments. In contrast, imipenem combined with vancomycin merely yielded weak additive or indifferent effects against 10 MDR isolates of Staphylococcus aureus, all of which were susceptible to imipenem.

  7. Synergistic hemolysins of coagulase-negative staphylococci (CoNS).

    Science.gov (United States)

    Różalska, Małgorzata; Derczyńska, Anna; Maszewska, Agnieszka

    2015-01-01

    A total of 104 coagulase negative staphylococci, belonging to S. capitis, S. hominis, S. haemolyticus and S. warneri, originating from the collection of the Department of Pharmaceutical Microbiology (ZMF), Medical University of Lodz, Poland, were tested for their synergistic hemolytic activity. 83% of strains produced δ-hemolysin, however, the percentage of positive strains of S. haemolyticus, S. warneri, S. capitis and S. hominis was different - 98%, 78%, 75% and 68%, respectively. Highly pure hemolysins were obtained from culture supernatants by protein precipitation with ammonium sulphate (0-70% of saturation) and extraction by using a mixture of organic solvents. The purity and molecular mass of hemolysins was determined by TRIS/Tricine PAGE. All CoNS hemolysins were small peptides with a molar mass of about 3.5 kDa; they possessed cytotoxic activity against the line of human foreskin fibroblasts ATCC Hs27 and lysed red cells from different mammalian species, however, the highest activity was observed when guinea pig, dog and human red blood cells were used. The cytotoxic effect on fibroblasts occurred within 30 minutes. The S. cohnii ssp. urealyticus strain was used as a control. The antimicrobial activity was examined using hemolysins of S. capitis, S. hominis, S. cohnii ssp. cohnii and S. cohnii ssp. urealyticus. Hemolysins of the two S. cohnii subspecies did not demonstrate antimicrobial activity. Cytolysins of S. capitis and S. hominis had a very narrow spectrum of action; out of 37 examined strains, the growth of only Micrococcus luteus, Corynebacterium diphtheriae and Pasteurella multocida was inhibited.

  8. Coagulase-negative Staphylococci in Conjunctivitis and Blepharitis

    Institute of Scientific and Technical Information of China (English)

    1993-01-01

    Coagulase-negative staphylococcus (C-NS) are regarded asnormal flora of the lids and conjunctiva.The ability of these organisms tocause conjunctivitis and blepharitis can be overlooked or disregarded.Toelucidate the role of individual C-NS species in these eye diseases wecompared Staphylococcus sp.isolated from the conjunctiva and lids of 50healthy volunteers with 248 strains of Staphylococcus isolated frompatients with staphylococcal conjunctivitis or blepharitis.S.epidermidiswas the most frequent spec...

  9. Cell surface hydrophobicity and charge of Staphylococcus aureus and coagulase-negative staphylococci from bovine mastitis.

    Science.gov (United States)

    Mamo, W; Rozgonyi, F; Brown, A; Hjertén, S; Wadström, T

    1987-03-01

    The effects of seven growth media on cell surface hydrophobicity of a collection of Staphylococcus aureus and coagulase-negative staphylococci isolated from bovine mastitis were compared in the salt-aggregation test. Thirty-three per cent of Staph. aureus strains showed extremely high cell surface hydrophobicity (auto-aggregated) and 28% were moderately hydrophobic while 26% were hydrophilic after growth on horse blood agar at 37 degrees C for 18 h. There were great variations in the proportion and degree of the hydrophobicity depending on the medium used. Cultivations on/in capsule-inducing media caused a shift from a high to a low degree of hydrophobicity, although a microscopically detectable capsule or slime layer was seen in only one strain. This strain and encapsulated reference strains had a hydrophilic cell surface and migrated faster in free zone electrophoresis than cells of unencapsulated strains. Cells of strains grown on staphylococcus medium 110 agar migrated faster than those grown on horse blood agar regardless of their capsule production. Coagulase-negative staphylococci showed uniformly hydrophilic cell surface after cultivation on horse blood agar, but not when grown in tryptic soy broth or proteose peptone broth. It was concluded that most of the Staph. aureus strains from bovine mastitis under a variety of growth conditions in stationary phase culture constantly expressed hydrophobic cell surface.

  10. Correlation between beta-lactamase production and MIC values against penicillin with coagulase negative staphylococci.

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    Narayani T

    1989-07-01

    Full Text Available Two hundred strains of coagulase negative staphylococci (CNS isolated from various clinical specimens (116 and healthy hospital personnel (84 were investigated for the production of beta-lactamases by means of three iodometric techniques and correlated with the minimum inhibitory concentration (MIC values of penicillin-G by agar dilution technique and disc diffusion technique. One hundred and fifty (75.0% of the 200 strains tested produced beta-lactamases. Seventy two per cent of the CNS were found to be beta-lactamase positive by the starch paper technique which was the most sensitive one in our study. The MIC values of penicillin against CNS ranged from less than or equal to 1.25 to greater than or equal to 2000 units. The present study indicated the higher prevalence of beta-lactamase producers with increased penicillin resistance among CNS strains isolated from healthy carriers and hospitalised patients.

  11. Significância clínica de estafilococos coagulase-negativa isolados de recém-nascidos Clinical significance of coagulase-negative staphylococci isolated from neonates

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    Maria de Lourdes R.S. Cunha

    2002-01-01

    Full Text Available Objetivo: avaliar a significância clínica de estafilococos coagulase-negativa (ECN isolados de processos infecciosos em recém-nascidos da unidade neonatal do Hospital das Clínicas da Faculdade de Medicina de Botucatu. Método: as linhagens de ECN isoladas foram identificadas e classificadas em significativas e contaminantes, com base em uma série de dados clínicos e laboratoriais obtidos dos prontuários dos pacientes internados na unidade neonatal. Foram pesquisados os dados referentes a fatores perinatais de risco para infecção, evolução clínica, alterações do hemograma e/ou positividade de proteína C-reativa e antibioticoterapia. Resultados: das 117 linhagens de ECN isoladas, 60 (51,3% foram classificadas como significativas, e 57 (48,7% como contaminantes. Das 54 crianças com infecção por ECN, 43 (79,6% eram prematuras, e 27 (50,0% tiveram peso ao nascimento Conclusões: a maioria dos recém-nascidos com infecção por ECN apresentou fatores predisponentes importantes para a instalação do processo infeccioso, incluindo o peso de nascimento Objective: to evaluate the clinical significance of coagulase-negative staphylococci (CNS isolated from newborns’ infections at Neonatal Unit of Hospital das Clínicas da Faculdade de Medicina de Botucatu. Methods: the CNS strains isolated were identified and classified as clinically significant and contaminant, based on a series of clinical and laboratory data obtained from patients who stayed in the Neonatal Unit. The following data were analyzed: risk factors for infections, clinical evolution, abnormal blood cell counts and/or C-reactive protein e antibiotic therapy. Results: among the 117 CNS strains isolated, 60 (51.3% were classified as significant and 57 (48.7% as contaminant. Among the 54 infants infected by CNS, 43 (79.6% presented very low birthweight (< 1,500g. Most of the infants infected by CNS were submitted to two or more invasive procedures (77.8%, including use of

  12. Antimicrobial resistance levels amongst staphylococci isolated from clinical cases of bovine mastitis in Kosovo.

    Science.gov (United States)

    Mehmeti, Ibrahim; Behluli, Behlul; Mestani, Mergim; Ademi, Arsim; Nes, Ingolf F; Diep, Dzung B

    2016-10-31

    Mastitis is one of the most frequent and costly disease in cattle. We studied milk samples from cattle with mastitis from farms in Kosovo to identify mastitis-causing pathogens and possible effective antibiotics. Our ultimate goal is to help implement adequate antibiotic management and treatment practices in Kosovo METHODOLOGY: A total of 152 milk samples were collected from cows with clinical mastitis from different farms in Kosovo. After identification of microorganisms, antibiotic susceptibility and the occurrence of enterotoxins was investigated. Staphylococci were found in 89 samples, of which 58 were coagulase negative and 31 coagulase positive. S. aureus was isolated from 27 samples, S. epidermidis from 25, and S. chromogenes from 15, while other species of staphylococci were isolated from the remaining 22 isolates. Interestingly, the bacterial diversity was different between cows in different periods of lactation and among different breeds. Most of the isolates (76/89) were resistant to two or more antibiotics. The highest resistance was to penicillin and ampicillin (> 65%), followed by tetracycline, oxacillin, streptomycin, chloramphenicol (> 23%), and less than 3% to erythromycin. Of the 89 isolates, 40 produced enterotoxins that were most frequently typed as A and C. We detected human bacterial pathogens in the cultures of milk samples from cows with mastitis. The isolates demonstrated resistance to two or more antibiotics, some of which are frequently used to treat animal and human infections. We recommend increased control and more stringent use of antibiotics in veterinary as well as human medicine.

  13. Detection of enterotoxins genes in coagulase-negative staphylococci isolated from foods Detecção de genes de enterotoxinas em estafilococos coagulase-negativa isolados de alimentos

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    Maria de Lourdes Ribeiro de Souza da Cunha

    2006-03-01

    Full Text Available Staphylococcal food poisoning is caused by ingestion of enterotoxins preformed in the food contaminated essentially through human manipulation or raw material obtained from animals. Although coagulase-positive Staphylococcus aureus is the main agent responsible for food intoxication, some researches emphasise that coagulase-negative staphylococci (CNS are able to produce staphylococcal enterotoxins and may be a potential cause of food poisoning. In the present study CNS were isolated from foods and the toxigenic capacity of the strains determined. A total of 88 food samples were analysed and 22.7% were positive for CNS strains. Staphylococcal counts ranged from 3.0 x 10² to 1.4 x 10(6 CFU/g or mL of food examined. S. epidermidis predominated among the isolates (40%. Further isolates included S. xylosus (20%, S. warneri (20%, S. saccharolyticus (15%, and S. hominis (5%. Four isolates were positive for enterotoxin genes, as detected by polymerase chain reaction, with sea being the predominant gene. Although no enterotoxin production was detected by the reverse passive latex agglutination method, the data showed that the toxigenic capacity of CNS should not be ignored, requiring investigation of this group of microorganisms in food.A intoxicação alimentar estafilocócica ocorre devido à ingestão de alimentos contaminados com enterotoxinas. Essa contaminação tem sido oriunda, principalmente, da manipulação humana, ou de matérias-primas procedentes de animais portadores. Embora Staphylococcus aureus coagulase positiva, seja o principal agente de intoxicação alimentar, alguns pesquisadores enfatizam que os estafilococos coagulase-negativa (ECN podem produzir as enterotoxinas estafilocócicas, podendo contribuir para a intoxicação alimentar. Este estudo teve como objetivos isolar os ECN de alimentos e verificar a capacidade enterotoxigênica dessas linhagens. Foram estudadas 88 amostras de alimentos, sendo que 22,7% foram positivas para

  14. Preliminary evaluation of a new clinical algorithm to interpret blood cultures growing coagulase-negative staphylococci.

    Science.gov (United States)

    Schnell, David; Lécuyer, Hervé; Geeraerts, Thomas; Dumenil, Anne-Sylvie; Bille, Emmanuelle; Mercier, Frédéric J; Benhamou, Dan; Zahar, Jean-Ralph

    2013-07-01

    Evaluating the clinical significance of blood cultures positive for coagulase-negative staphylococci (CoNS) is of critical importance since these microorganisms represent both the first contaminants of blood cultures and one of the leading causes of bloodstream infection (BSI). This prospective 2-centre study aimed to compare a previously reported algorithm to a clinical algorithm based on our experience. We identified 84 patients with CoNS-positive blood cultures. Twenty-seven (32%) were considered to have BSI according to our study algorithm. Thirty-seven (44%) patients were considered to have CoNS BSI according to the previously reported algorithm. The 2 algorithms isolated patients with similar rates of recurrences and hospital mortality. Our algorithm seemed to result in less diagnoses of CoNS BSI without harmful consequences compared to the previously reported algorithm. The impact on patient outcome and the inappropriate use of antibiotics deserves further investigation.

  15. Antibiotic resistance profiles of coagulase-negative staphylococci in livestock environments.

    Science.gov (United States)

    Schoenfelder, Sonja M K; Dong, Ying; Feßler, Andrea T; Schwarz, Stefan; Schoen, Christoph; Köck, Robin; Ziebuhr, Wilma

    2017-02-01

    Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) have globally emerged in animal husbandry. In addition to methicillin resistance, LA-MRSA may carry a variety of novel and uncommon antimicrobial resistance genes. Occurrence of the same resistance genes in coagulase-negative staphylococci (CoNS) and S. aureus suggests an ongoing genetic exchange between LA-MRSA and other staphylococci whose driving forces in the ecological niche of the farm environment are, however, still poorly understood. To assess the potential of CoNS as putative reservoirs for antibiotic resistance genes, we analysed the antimicrobial susceptibility of CoNS from dust and manure samples obtained in 41 pig farms in Germany, most of them (36 of 41) with a proven LA-MRSA/MSSA history. Among the 344 isolates analysed, 18 different CoNS species were identified and S. sciuri represented the most prevalent species (46%). High resistance rates were detected for tetracycline (71%), penicillin (65%) and oxacillin (64%) as well as fusidic acid (50%), which was mainly due to reduced susceptibility among S. sciuri isolates. S. sciuri exhibited pronounced multiresistance, and many isolates were characterised by the carriage of a number of uncommon (multi)resistance genes (e.g. cfr, apmA, fexA) and decreased susceptibility towards last resort antibiotics such as linezolid and daptomycin. The combined data suggest that S. sciuri harbours a significant resistance gene pool that requires further attention. We hypothesise that members of this species, due to their flexible lifestyle, might contribute to the spread of such genes in livestock environments. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Bacteriocins of Non-aureus Staphylococci Isolated from Bovine Milk.

    Science.gov (United States)

    Carson, Domonique A; Barkema, Herman W; Naushad, Sohail; De Buck, Jeroen

    2017-09-01

    Non-aureus staphylococci (NAS), the bacteria most commonly isolated from the bovine udder, potentially protect the udder against infection by major mastitis pathogens due to bacteriocin production. In this study, we determined the inhibitory capability of 441 bovine NAS isolates (comprising 26 species) against bovine Staphylococcus aureus Furthermore, inhibiting isolates were tested against a human methicillin-resistant S. aureus (MRSA) isolate using a cross-streaking method. We determined the presence of bacteriocin clusters in NAS whole genomes using genome mining tools, BLAST, and comparison of genomes of closely related inhibiting and noninhibiting isolates and determined the genetic organization of any identified bacteriocin biosynthetic gene clusters. Forty isolates from 9 species (S. capitis, S. chromogenes, S. epidermidis, S. pasteuri, S. saprophyticus, S. sciuri, S. simulans, S. warneri, and S. xylosus) inhibited growth of S. aureus in vitro, 23 isolates of which, from S. capitis, S. chromogenes, S. epidermidis, S. pasteuri, S. simulans, and S. xylosus, also inhibited MRSA. One hundred five putative bacteriocin gene clusters encompassing 6 different classes (lanthipeptides, sactipeptides, lasso peptides, class IIa, class IIc, and class IId) in 95 whole genomes from 16 species were identified. A total of 25 novel bacteriocin precursors were described. In conclusion, NAS from bovine mammary glands are a source of potential bacteriocins, with >21% being possible producers, representing potential for future characterization and prospective clinical applications.IMPORTANCE Mastitis (particularly infections caused by Staphylococcus aureus) costs Canadian dairy producers $400 million/year and is the leading cause of antibiotic use on dairy farms. With increasing antibiotic resistance and regulations regarding use, there is impetus to explore bacteriocins (bacterially produced antimicrobial peptides) for treatment and prevention of bacterial infections. We

  17. Detecção do gene mecA em estafilococos coagulase negativa resistentes à oxacilina isolados da saliva de profissionais da enfermagem Detection of mecA gene in oxacillin-resistant coagulase-negative staphylococci isolated from the saliva of nursing professionals

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    Juliana de Oliveira Rosa

    2009-08-01

    Full Text Available Estafilococos coagulase negativa estão frequentemente associados às infecções nosocomiais e os profissionais da saúde podem ser reservatório e dissemina-los no hospital e comunidade. O objetivo desse estudo foi identificar espécies de estafilococos coagulase negativa isolados da saliva de profissionais da enfermagem, determinar o perfil de resistência e detectar o gene mecA. Foram selecionados 100 estafilococos coagulase negativa, sendo 41 identificados como Staphylococcus epidermidis, 25 Staphylococcus saprophyticus, 18 Staphylococcus haemolyticus, 8 Staphylococcus cohnii, 4 Staphylococcus lugdunenses, 3 Staphylococcus capitis, e 1 Staphylococcus Simulans. Desses, 32% apresentaram resistência à oxacilina, 84,4% à mupirocina, 32% à cefoxitina, e todos sensíveis a vancomicina. Dos estafilococos coagulase negativa resistentes à oxacilina, 93,7% desenvolveram-se no agar oxacilina (6µg/ml e o gene mecA foi detectado em 75%. Os resultados sinalizam que maiores investimentos devem ser direcionados a identificação das espécies de estafilococos coagulase negativa nas instituições de saúde e na comunidade.Coagulase-negative staphylococci are frequently associated with nosocomial infections, and healthcare professionals can be reservoirs and spread them in hospitals and in the community. The aim of this study was to identify species of coagulase-negative staphylococci isolated from the saliva of nursing professionals, determine the resistance profile and detect the mecA gene. One hundred coagulase-negative staphylococci were selected: 41 were identified as Staphylococcus epidermidis, 25 as Staphylococcus saprophyticus, 18 as Staphylococcus haemolyticus, eight as Staphylococcus cohnii, four as Staphylococcus lugdunenses, three as Staphylococcus capitis and one as Staphylococcus simulans. Of these, 32% presented oxacillin resistance, 84.4% mupirocin resistance and 32% cefoxitin resistance, and all were vancomycin sensitive. Among the

  18. Coagulase-negative Staphylococcus isolated from bloodculture: Causes or contaminants?

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    Stojanović Predrag

    2008-01-01

    Full Text Available Introduction According to the results of different investigations, the opinion that isolate of coagulase-negative staphylococci by bloodcultures represents the blood infection in 10-12% patient. The aim of the work was to determine the number of patients with blood infection-sepsa induced coagulase-negative staphylococci isolated by bloodcultures. Material and methods: The research was done at the Institute for Public Health in Niš. The basic group consisted of 56 patients and coagulase-negative staphylococci were segregated form their bloodcultures. The growth of microorganisms was monitored by the computerized apparatus 'Bactec 9120'. Coagulase-negative staphylococci were identified by the standard microbiologic method and Vitec AMS system. Results Eight patients (14.28% were found to have minimum two signs of blood infection. S. epidermis was isolated in four patients. S. saprophyticus was isolated in the patients on hemodialysis with implanted urinary catheter. S. capitis was isolated in the patients with infarctus miocardi. S. auricularis was isolated in child who was administered the immunosuppressive therapy before and during hospitalization due to an acute asthmatic attack. The isolate of S. simulans was cultivated from samples of the patients hospitalized due to the febrile state. Discussion A correct interpretation of coagulase-negative staphylococci findings in bloodcultures is an overriding part of medical treatment. The best laboratory indicators of the presence of coagulase-negative staphylococci in blood would be to segregate those of the same kind from two samples at the same time but from different anatomic sites in the presence of clinical signs of blood infection. Conclusion In the investigation conducted at the Public Health Institute-Nis, we determined that bloodcultures isolated coagulase-negative staphylococci represent the blood infection on 14.28% patient.

  19. An unusual presentation of sepsis caused by coagulase-negative staphylococci

    NARCIS (Netherlands)

    Nieboer, P; Gietema, JA; de Vries-Hospers, HG; van der Graaf, WTA

    1999-01-01

    We describe a 25-year-old male presenting with fever during the non-neutropenic phase of chemotherapy. The presentation was that of a viral infection. The cause of the fever turned out to be a bacteremia with coagulase-negative staphylococci (CONS) originating from a totally implanted venous access

  20. Persistence of clones of coagulase-negative staphylococci among premature neonates in neonatal intensive care units: two-center study of bacterial genotyping and patient risk factors

    NARCIS (Netherlands)

    C.L. Vermont (Clementien); N.G. Hartwig (Nico); A. Fleer; P. de Man (Peter); R. de Groot (Ronald); A.F. van Belkum (Alex); H.A. Verbrugh (Henri); J.N. van den Anker (John)

    1998-01-01

    textabstractFrom 1 January 1995 until 1 January 1996, we studied the molecular epidemiology of blood isolates of coagulase-negative staphylococci (CoNS) in the Neonatal Intensive Care Units (NICUs) of the Sophia Children's Hospital (SCH; Rotterdam, The Netherlands) and the Wilhelmi

  1. Emerging of coagulase negative staphylococci as a cause of mastitis in dairy animals: An environmental hazard

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    Jakeen K. El-Jakee

    2013-12-01

    Full Text Available In Egypt, knowledge about the coagulase negative staphylococci (CNS involved in mastitic animals is limited. CNS have emerged to be pathogens causing intramammary infections in Egyptian dairy herds. Therefore, the current study was conducted to investigate the occurrence of CNS in dairy ruminants (cattle, buffaloes, sheep and goats. A total of 884 quarter milk samples were investigated to study the prevalence of CNS among mastitic and subclinically mastitic cows, buffalo–cows, ewes and does in Egypt. Identification of the isolates was achieved using API staph test and polymerase chain reaction (PCR. CNS were isolated from the examined subclinical mastitic cattle, buffaloes, sheep and goats with percentages of 16.6%, 59.4%, 50% and 55.6%, respectively. Staphylococcus xylosus, Staphylococcus cohnii, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus saprophyticus, Staphylococcus chromogenes, Staphylococcus lentus, Staphylococcus lugdunensis and Staphylococcus simulans were identified as CNS that recovered from the examined milk samples. The CNS as mastitis-causing agents could not be neglected as they can cause substantial economic losses.

  2. The effects of four different drugs administered through catheters on slime production in coagulase negative Staphylococci

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    J. Sedef Göçmen

    2012-12-01

    Full Text Available Objectives: Higher rate of slime production has been found in pathogen bacteria strains. Accordingly, the factors thatcontribute to higher slime production rate increase the infection risk, while the factors that reduce the slime productionrate will reduce the infection risk. The effect of some drugs that are administered through catheters in intensive careunits on slime production with coagulase negative Staphylococci was investigated.Materials and methods: In this study, the effect of four different preparations containing Glyceryl trinitrate (Perlinganit®, Dexmedetomidine (Precedex®, Esmolol (Brevibloc®, and Propofol (Propofol® on slime production of 24Staphylococcus epidermidis strains isolated from blood cultures of patients, and reference strain were investigated. Slimeproduction was determined using ‘the quantitative microdilution plaque test’ described by Christensen.Results: Under controlled medium, eight strains formed slimes, and in the media containing esmolol, glyceryl trinitrate,dexmedetomidine, and propofol slimes were positive for five, 21, 15, and 18 strains, respectively. The rate of slime productionin glyceryl trinitrate, dexmedetomidine, and propofol containing media were higher than that of the controls.Conclusions: In the light of the results of this study, it is concluded that the drugs and/or additives increase the rate ofslime production. The effects of the preparations administered through catheters on slime production should be investigated,and these effects should be kept in mind during their use. J Microbiol Infect Dis 2012; 2(4: 150-154Key words: Slime Production, Coagulase Negative Staphyloccoci, Parenteral drugs

  3. Impact of Coagulase-Negative Staphylococci and Other Germs on Sperm Forms

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    Ricardo Lozano-Hernández

    2017-08-01

    Full Text Available Coagulase-negative Staphylococci (CoNS is part of the microbiota of the male genitourinary tract, sometimes it has been considered as possible pathogenic microorganism. In the 5th version of sperm manual (WHO, 2010 sperm morphology criterion is very restricted to 4% of normal heads whereas David’s criterion evaluates several spermatic forms. The abnormalities of sperm forms were evaluated according to criteria of spermatic morphology: WHO and David in semen samples with bacterial concentrations ≤ 103, 104 and ≥ 105 CFU/mL of CoNS as of other bacteria. Spermogram, sperm culture and antibodies anti-Chlamydia trachomatis IgA detection in 281 semen samples men were performed. CoNS was the most frequent germ isolated in pure culture (9.25%. Semen samples with CoNS showed higher round cells and microcephalus forms by means of David’s criterion. CoNS in higher concentrations than 104 CFU/mL may have a negative impact on sperm cellularity, sperm head and probably on fertility.

  4. Antimicrobial Resistance Profile of Planktonic and Biofilm Cells of Staphylococcus aureus and Coagulase-Negative Staphylococci

    Science.gov (United States)

    de Oliveira, Adilson; Cataneli Pereira, Valéria; Pinheiro, Luiza; Moraes Riboli, Danilo Flávio; Benini Martins, Katheryne; Ribeiro de Souza da Cunha, Maria de Lourdes

    2016-01-01

    The objective of the present study was to determine the antimicrobial resistance profile of planktonic and biofilm cells of Staphylococcus aureus and coagulase-negative staphylococci (CoNS). Two hundred Staphylococcus spp. strains were studied, including 50 S. aureus and 150 CoNS strains (50 S. epidermidis, 20 S. haemolyticus, 20 S. warneri, 20 S. hominis, 20 S. lugdunensis, and 20 S. saprophyticus). Biofilm formation was investigated by adherence to polystyrene plates. Positive strains were submitted to the broth microdilution method to determine the minimum inhibitory concentration (MIC) for planktonic and biofilm cells and the minimal bactericidal concentration for biofilm cells (MBCB). Forty-nine Staphylococcus spp. strains (14 S. aureus, 13 S. epidermidis, 13 S. saprophyticus, 3 S. haemolyticus, 1 S. hominis, 3 S. warneri, and 2 S. lugdunensis) were biofilm producers. These isolates were evaluated regarding their resistance profile. Determination of planktonic cell MIC identified three (21.4%) S. aureus strains that were resistant to oxacillin and six (42.8%) that were resistant to erythromycin. Among the CoNS, 31 (88.6%) strains were resistant to oxacillin, 14 (40%) to erythromycin, 18 (51.4%) to gentamicin, and 8 (22.8%) to sulfamethoxazole/trimethoprim. None of the planktonic isolates were resistant to vancomycin or linezolid. MICs were 2-, 4-, 8-, and up to 16-fold higher for biofilm cells than for planktonic cells. This observation was more common for vancomycin and erythromycin. The MBCB ranged from 8 to >256 µg/mL for oxacillin, 128 to >128 µg/mL for vancomycin, 256 to >256 µg/mL for erythromycin and gentamicin, >64 µg/mL for linezolid, and 32/608 to >32/608 µg/mL for sulfamethoxazole/trimethoprim. The results showed considerably higher MICs for S. aureus and CoNS biofilm cells compared to planktonic cells. Analysis of MBCM confirmed that even high concentrations of vancomycin were unable to eliminate the biofilms of S. aureus and CoNS species

  5. Antimicrobial Resistance Profile of Planktonic and Biofilm Cells of Staphylococcus aureus and Coagulase-Negative Staphylococci.

    Science.gov (United States)

    de Oliveira, Adilson; Cataneli Pereira, Valéria; Pinheiro, Luiza; Moraes Riboli, Danilo Flávio; Benini Martins, Katheryne; Ribeiro de Souza da Cunha, Maria de Lourdes

    2016-09-01

    The objective of the present study was to determine the antimicrobial resistance profile of planktonic and biofilm cells of Staphylococcus aureus and coagulase-negative staphylococci (CoNS). Two hundred Staphylococcus spp. strains were studied, including 50 S. aureus and 150 CoNS strains (50 S. epidermidis, 20 S. haemolyticus, 20 S. warneri, 20 S. hominis, 20 S. lugdunensis, and 20 S. saprophyticus). Biofilm formation was investigated by adherence to polystyrene plates. Positive strains were submitted to the broth microdilution method to determine the minimum inhibitory concentration (MIC) for planktonic and biofilm cells and the minimal bactericidal concentration for biofilm cells (MBCB). Forty-nine Staphylococcus spp. strains (14 S. aureus, 13 S. epidermidis, 13 S. saprophyticus, 3 S. haemolyticus, 1 S. hominis, 3 S. warneri, and 2 S. lugdunensis) were biofilm producers. These isolates were evaluated regarding their resistance profile. Determination of planktonic cell MIC identified three (21.4%) S. aureus strains that were resistant to oxacillin and six (42.8%) that were resistant to erythromycin. Among the CoNS, 31 (88.6%) strains were resistant to oxacillin, 14 (40%) to erythromycin, 18 (51.4%) to gentamicin, and 8 (22.8%) to sulfamethoxazole/trimethoprim. None of the planktonic isolates were resistant to vancomycin or linezolid. MICs were 2-, 4-, 8-, and up to 16-fold higher for biofilm cells than for planktonic cells. This observation was more common for vancomycin and erythromycin. The MBCB ranged from 8 to >256 µg/mL for oxacillin, 128 to >128 µg/mL for vancomycin, 256 to >256 µg/mL for erythromycin and gentamicin, >64 µg/mL for linezolid, and 32/608 to >32/608 µg/mL for sulfamethoxazole/trimethoprim. The results showed considerably higher MICs for S. aureus and CoNS biofilm cells compared to planktonic cells. Analysis of MBCM confirmed that even high concentrations of vancomycin were unable to eliminate the biofilms of S. aureus and CoNS species

  6. Antimicrobial Resistance Profile of Planktonic and Biofilm Cells of Staphylococcus aureus and Coagulase-Negative Staphylococci

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    Adilson de Oliveira

    2016-09-01

    Full Text Available The objective of the present study was to determine the antimicrobial resistance profile of planktonic and biofilm cells of Staphylococcus aureus and coagulase-negative staphylococci (CoNS. Two hundred Staphylococcus spp. strains were studied, including 50 S. aureus and 150 CoNS strains (50 S. epidermidis, 20 S. haemolyticus, 20 S. warneri, 20 S. hominis, 20 S. lugdunensis, and 20 S. saprophyticus. Biofilm formation was investigated by adherence to polystyrene plates. Positive strains were submitted to the broth microdilution method to determine the minimum inhibitory concentration (MIC for planktonic and biofilm cells and the minimal bactericidal concentration for biofilm cells (MBCB. Forty-nine Staphylococcus spp. strains (14 S. aureus, 13 S. epidermidis, 13 S. saprophyticus, 3 S. haemolyticus, 1 S. hominis, 3 S. warneri, and 2 S. lugdunensis were biofilm producers. These isolates were evaluated regarding their resistance profile. Determination of planktonic cell MIC identified three (21.4% S. aureus strains that were resistant to oxacillin and six (42.8% that were resistant to erythromycin. Among the CoNS, 31 (88.6% strains were resistant to oxacillin, 14 (40% to erythromycin, 18 (51.4% to gentamicin, and 8 (22.8% to sulfamethoxazole/trimethoprim. None of the planktonic isolates were resistant to vancomycin or linezolid. MICs were 2-, 4-, 8-, and up to 16-fold higher for biofilm cells than for planktonic cells. This observation was more common for vancomycin and erythromycin. The MBCB ranged from 8 to >256 µg/mL for oxacillin, 128 to >128 µg/mL for vancomycin, 256 to >256 µg/mL for erythromycin and gentamicin, >64 µg/mL for linezolid, and 32/608 to >32/608 µg/mL for sulfamethoxazole/trimethoprim. The results showed considerably higher MICs for S. aureus and CoNS biofilm cells compared to planktonic cells. Analysis of MBCM confirmed that even high concentrations of vancomycin were unable to eliminate the biofilms of S. aureus and Co

  7. Automated systems in the identification and determination of methicillin resistance among coagulase negative staphylococci

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    Juliana Caierão

    2006-05-01

    Full Text Available Coagulase-negative staphylococci (CoNS are an important cause of nosocomial bacteremia, specially in patients with indwelling devices or those submitted to invasive medical procedures. The identification of species and the accurate and rapid detection of methicillin resistance are directly dependent on the quality of the identification and susceptibility tests used, either manual or automated. The objective of this study was to evaluate the accuracy of two automated systems MicroScan and Vitek - in the identification of CoNS species and determination of susceptibility to methicillin, considering as gold standard the biochemical tests and the characterization of the mecA gene by polymerase chain reaction, respectively. MicroScan presented better results in the identification of CoNS species (accuracy of 96.8 vs 78.8%, respectively; isolates from the following species had no precise identification: Staphylococcus haemolyticus, S. simulans, and S. capitis. Both systems were similar in the characterization of methicillin resistance. The higher discrepancies for gene mec detection were observed among species other than S. epidermidis (S. hominis, S. saprophyticus, S. sciuri, S. haemolyticus, S. warneri, S. cohnii, and those with borderline MICs.

  8. [Relatedness of methicillin-resistant coagulase-negative staphylococci].

    Science.gov (United States)

    Bogiel, Tomasz; Mikucka, Agnieszka; Deptuła, Aleksander; Gospodarek, Eugenia

    2009-01-01

    Many identification and relatedness studies methods had been commonly used for epidemiological studies in microbiological laboratories. Apart from phenotypic methods, genotypic are also often used. The aim of this study was to compare, obtained by PFGE chromosomal DNA patterns of methicillin-resistant S. epidermidis strains isolated from clinical material. 46 methicillin-resistant S. epidermidis strains were included in this study. Most of them were isolated from wound swabs (65.2%) and catheters (19.6%) from different surgical clinics (76.1%). To identify strains and receive biochemical profiles, ID 32 Staph tests and GPI cards of Vitek 1 system were used. Pulsed-field gel electrophoresis and Tenover et al. interpretation were used to compare chromosomal DNA patterns of examined strains. 44 and 42 PFGE patterns of chromosomal DNA were received, using visual interpretation classifying two pairs of strains as the same, two pairs as closely related and three pairs as probably related. Strains classified as identical and similar in visual evaluation were indistinguishable in Molecular Analyst DST interpretation, probably due to tolerance in bands location pattern. Strains probably related in visual interpretation represent at least 96% similarity in Molecular Analyst DST but different susceptibility and biochemical profiles obtained by ID 32 Staph and Vitek 1. PFGE analysis had foremost capacity to distinguish methicillin-resistant S. epidermidis strains using visual interpretation and Molecular Analyst DST (Bio-Rad) program and seems to be useful method in epidemiological studies. Strains with the same PFGE pattern, had different susceptibility and biochemical profiles.

  9. Toxigenicity in Staphylococcus aureus and Coagulase-Negative Staphylococci: Epidemiological and Molecular Aspects

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    M.L.R.S. Cunha

    2008-01-01

    Full Text Available Representatives of the Staphylococcus genus are the most common pathogens found in hospital environments, and they are etiological agents for a large variety of infections. Various virulence factors are responsible for the symptoms and severity of infections caused by Staphylococcus aureus. Among them are staphylococcal enterotoxins (SEs, which cause staphylococcal food poisoning, and toxic shock syndrome toxin-1 (TSST-1. Some reports indicate that TSST-1 and staphylococcal enterotoxins are also produced by coagulase-negative staphylococci (CNS. The present review aimed to discuss general aspects of staphylococcal toxins as well as the epidemiology, genetics and detection of toxins in Staphylococcus aureus and coagulase-negative staphylococci, since these microorganisms are becoming more and more frequent in nosocomial infections.

  10. Activity of carvacrol against Coagulase-negative Staphylococci biofilm related infections

    OpenAIRE

    Lima, Cláudia Sofia Afonso de

    2015-01-01

    Dissertação de mestrado em Bioengenharia Coagulase-negative staphylococci (CoNS) are frequently found in healthy human skin and mucosae, and hence, have long been considered non-pathogenic bacteria. However, in recent years, they have been recognized as important etiological agents of healthcareassociated infections, being particularly associated with patients with indwelling medical devices. Despite their importance as nosocomial pathogens, very little is known regarding th...

  11. Finding of coagulase negative staphylococci in the herd with an increased number of somatic cells in milk and their antimicrobial susceptibility

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    Katić Vera

    2013-01-01

    Full Text Available Coagulase-negative staphylococci (CNS are generally considered to be opportunistic pathogens. Controlling CNS mastitis is difficult because the epidemiology is not clear, and the CNS group consists of about 40 different Staphylococcus species. Therefore, the aim of this study was to determine the prevalence of coagulasenegative staphylococci in milk of the cows with subclinical mastitis, as well as to determine different CNS species isolated from quarter milk samples for their susceptibility to antimicrobials used commonly for mastitis therapy. On the farm where there was found an increase of somatic cells in bulk milk, 112 dairy cows were examined by mastitis test. From 52 udder quarters where mastitis test showed an increase of somatic cells, milk samples were taken for bacteriological examination. For isolating the causes of mastitis there was used blood agar. Identification of the causative agents of mastitis was carried out on the basis of colony appearance on blood agar and their physiological characteristics. Coagulasepositive staphylococci sensitivity which cause mastitis was tested by Kirby Bauer method. For susceptibility testing there were used commercially produced discs containing: 10 IU penicillin, amoxicillin/clavulanic acid (20 +10 μg, cloxacillin 25 μg, 30 μg amoxicillin, cephalexin 30 μg, ceftiofur 30 μg, 15 μg lincomycin, gentamicin and tetracycline 30 μg. The sensitivity of microorganisms was evaluated on the basis of inhibition zone diameter recommended by the manufacturer and was labeled as sensitive (S moderately sensitive (I or resistant (R. Coagulase-negative staphylococci were isolated from 61.53% of samples from cows with subclinical mastitis, making them the most common cause of subclinical mastitis. The highest resistance of coagulase-negative staphylococci was found to penicillin G (58.33% of isolates. Full sensitivity of coagulase-negative staphylococci was found to amoxicillin / clavulanic acid (100% of

  12. Microbial Diversity in Milk of Women With Mastitis: Potential Role of Coagulase-Negative Staphylococci, Viridans Group Streptococci, and Corynebacteria.

    Science.gov (United States)

    Mediano, Pilar; Fernández, Leonides; Jiménez, Esther; Arroyo, Rebeca; Espinosa-Martos, Irene; Rodríguez, Juan M; Marín, María

    2017-05-01

    Lactational mastitis constitutes a significant cause of premature weaning. However, its etiology, linked to the presence of pathogenic microorganisms, has been scarcely reported. Research aim: The aim of this study was to describe the microbial diversity in milk samples from women suffering from lactational mastitis and to identify more accurately a collection of isolates belonging to coagulase-negative staphylococci, streptococci, and coryneform bacteria. This is a cross-sectional descriptive one-group study. A total of 5,009 isolates from 1,849 mastitis milk samples was identified by culture, biochemical, and/or molecular methods at the species or genus level. A more precise identification of a collection of 211 isolates was carried out by 16S rRNA gene sequencing. Mean total bacterial count in milk samples was 4.11 log10 colony-forming units/ml, 95% confidence interval [4.08, 4.15]. Staphylococcus epidermidis was the most common species being isolated from 91.56% of the samples, whereas Staphylococcus aureus was detected in 29.74%. Streptococci and corynebacteria constituted the second (70.20%) and third (16.60%) most prevalent bacterial groups, respectively, found in this study. In contrast, Candida spp. was present in only 0.54% of the samples. Sequencing of the 16S rRNA gene revealed a high diversity of bacterial species among identified isolates. Many coagulase-negative staphylococci, viridans group streptococci, and corynebacteria, usually dismissed as contaminant bacteria, may play an important role as etiologic agents of mastitis. Proper diagnosis of mastitis should be established after performing microbiological testing of milk based on standardized procedures. A reliable analysis must identify the mastitis-causing pathogen(s) at the species level and its(their) concentration(s).

  13. BACTERIOLOGICAL STUDY OF COAGULASE-POSITIVE AND COAGULASE-NEGATIVE STAPHYLOCOCCI IN RELATION TO METHICILLIN SENSITIVITY TESTING

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    Padmanabham Yalangi

    2016-10-01

    Full Text Available BACKGROUND Staphylococcus aureus has long been recognised as an important pathogen in human disease. Staphylococci infection occurs regularly in hospitalised patients and has serious consequences despite antibiotic therapy. Shortly after introduction of methicillin after clinical use Methicillin-Resistant Staphylococcus Aureus (MRSA were identified in many countries and become one of the most common causes of nosocomial infections. The aim of the study is to know the methicillin sensitivity of both coagulase-negative and coagulase-positive staphylococci isolated from various samples. MATERIALS AND METHODS 100 strains of staphylococci both coagulase positive and coagulase negative were isolated in the Department of Microbiology from various clinical samples. They were confirmed by morphology, staining methods and by using standard bacteriological procedures and biochemical reactions. Antibiotic susceptibility testing was performed by Kirby Bauer disc diffusion test. RESULTS Predominant species from pus were S. epidermidis (42.42% and from sputum S. haemolyticus (31.81% from blood S. haemolyticus (53.33%. 53% of strains produced beta-lactamase. Majority 47.22% by S. epidermidis from pus followed by S. haemolyticus 23.33% from pus. Beta-lactamase production was least from throat swab (5.55%. Out of 32 coagulase-positive staphylococci tested to methicillin 15 (46.87% were found to be sensitive, 17 (53.13% were found to be resistant. Out of 68 coagulase-negative staphylococci tested, 13 (19.11% were found to sensitive and 55 (80.88% were found to be resistant. 72% of strains were sensitive to novobiocin and 28% resistant to novobiocin. 43% showed drug resistance to 2 drugs. 14% to 3 drugs and 5 drugs. 6% of staphylococci sensitive to all the 10 drugs. CONCLUSION MRSA is a type of bacteria that is resistant to a number of widely used antibiotics. This means MRSA infections can be more difficult to treat than other bacterial infections. In recent years

  14. 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.

  15. Reduced expression of PBP-2A by neonatal mecA-positive coagulase-negative staphylococci (CoNS) blood isolates : beta-lactams are useful first-line agents for the treatment of neonatal CoNS sepsis, restricting the use of vancomycin

    NARCIS (Netherlands)

    Fleer, Andre; Hemels, Marieke A. C.; Paauw, Armand; Krediet, Tannette G.

    2012-01-01

    Vancomycin use for neonatal coagulase-negative staphylococci (CoNS) sepsis is based on a high CoNS carriage rate of mecA, encoding penicillin-binding protein (PBP)-2a, with low affinity for, and associated with resistance to, -lactam antibiotics. The relationship between mecA gene carriage, phenotyp

  16. Reduced expression of PBP-2A by neonatal mecA-positive coagulase-negative staphylococci (CoNS) blood isolates : beta-lactams are useful first-line agents for the treatment of neonatal CoNS sepsis, restricting the use of vancomycin

    NARCIS (Netherlands)

    Fleer, Andre; Hemels, Marieke A. C.; Paauw, Armand; Krediet, Tannette G.

    2012-01-01

    Vancomycin use for neonatal coagulase-negative staphylococci (CoNS) sepsis is based on a high CoNS carriage rate of mecA, encoding penicillin-binding protein (PBP)-2a, with low affinity for, and associated with resistance to, -lactam antibiotics. The relationship between mecA gene carriage, phenotyp

  17. Genotypic Diversity of Coagulase-Negative Staphylococci Causing Endocarditis: a Global Perspective▿

    Science.gov (United States)

    Petti, Cathy A.; Simmon, Keith E.; Miro, Jose M.; Hoen, Bruno; Marco, Francesc; Chu, Vivian H.; Athan, Eugene; Bukovski, Suzana; Bouza, Emilio; Bradley, Suzanne; Fowler, Vance G.; Giannitsioti, Efthymia; Gordon, David; Reinbott, Porl; Korman, Tony; Lang, Selwyn; Garcia-de-la-Maria, Cristina; Raglio, Annibale; Morris, Arthur J.; Plesiat, Patrick; Ryan, Suzanne; Doco-Lecompte, Thanh; Tripodi, Francesca; Utili, Riccardo; Wray, Dannah; Federspiel, J. Jeffrey; Boisson, K.; Reller, L. Barth; Murdoch, David R.; Woods, Christopher W.

    2008-01-01

    Coagulase-negative staphylococci (CNS) are important causes of infective endocarditis (IE), but their microbiological profiles are poorly described. We performed DNA target sequencing and susceptibility testing for 91 patients with definite CNS IE who were identified from the International Collaboration on Endocarditis—Microbiology, a large, multicenter, multinational consortium. A hierarchy of gene sequences demonstrated great genetic diversity within CNS from patients with definite endocarditis that represented diverse geographic regions. In particular, rpoB sequence data demonstrated unique genetic signatures with the potential to serve as an important tool for global surveillance. PMID:18367572

  18. Genotypic diversity of coagulase-negative staphylococci causing endocarditis: a global perspective.

    Science.gov (United States)

    Petti, Cathy A; Simmon, Keith E; Miro, Jose M; Hoen, Bruno; Marco, Francesc; Chu, Vivian H; Athan, Eugene; Bukovski, Suzana; Bouza, Emilio; Bradley, Suzanne; Fowler, Vance G; Giannitsioti, Efthymia; Gordon, David; Reinbott, Porl; Korman, Tony; Lang, Selwyn; Garcia-de-la-Maria, Cristina; Raglio, Annibale; Morris, Arthur J; Plesiat, Patrick; Ryan, Suzanne; Doco-Lecompte, Thanh; Tripodi, Francesca; Utili, Riccardo; Wray, Dannah; Federspiel, J Jeffrey; Boisson, K; Reller, L Barth; Murdoch, David R; Woods, Christopher W

    2008-05-01

    Coagulase-negative staphylococci (CNS) are important causes of infective endocarditis (IE), but their microbiological profiles are poorly described. We performed DNA target sequencing and susceptibility testing for 91 patients with definite CNS IE who were identified from the International Collaboration on Endocarditis-Microbiology, a large, multicenter, multinational consortium. A hierarchy of gene sequences demonstrated great genetic diversity within CNS from patients with definite endocarditis that represented diverse geographic regions. In particular, rpoB sequence data demonstrated unique genetic signatures with the potential to serve as an important tool for global surveillance.

  19. Genetic Diversity and Antibiotic Resistance Among Coagulase-Negative Staphylococci Recovered from Birds of Prey in Portugal.

    Science.gov (United States)

    Sousa, Margarida; Silva, Nuno; Igrejas, Gilberto; Sargo, Roberto; Benito, Daniel; Gómez, Paula; Lozano, Carmen; Manageiro, Vera; Torres, Carmen; Caniça, Manuela; Poeta, Patrícia

    2016-12-01

    Wild animal populations in contact with antimicrobials and antimicrobial resistant bacteria that are daily released into the environment are able to become unintentional hosts of these resistant microorganisms. To clarify this issue, our study evaluated the presence of antibiotic resistance determinants on coagulase-negative staphylococci recovered from birds of prey and studied their genetic relatedness by pulsed-field gel electrophoresis (PFGE). The unusual vga(A) and erm(T) genes, which confer resistance to clindamycin and erythromycin, respectively, were detected in Staphylococcus sciuri or Staphylococcus xylosus strains and the tet(K) gene in Staphylococcus kloosii. The PFGE patterns showed that three S. xylosus (isolated of Strix aluco and Otus scops) and two S. sciuri (recovered from Strix aluco and Milvus migrans) were clonally indistinguishable. These animals could be a source of unusual antimicrobial resistance determinants for highly used antibiotics in veterinary clinical practice.

  20. Detection of Methicillin-Resistant Staphylococci Isolated from Food Producing Animals: A Public Health Implication

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    Etinosa O. Igbinosa

    2016-07-01

    Full Text Available The emergence of antibiotic-resistant bacteria in food animals is a potential public health concern. Staphylococci are a significant opportunistic pathogen both in humans and dairy cattle. In the present study, the genotypic characterization of methicillin-resistant staphylococcal strains recovered from dairy cattle in a rural community (Okada, Edo State, Nigeria was investigated. A total of 283 samples from cattle (137 milk samples and 146 nasal swabs were assessed between February and April 2015. Antimicrobial susceptibility was performed by Kirby-Bauer disc diffusion method. Polymerase chain reaction (PCR assay was employed for the detection of 16S rRNA, mecA and Panton-Valentine Leucocidinis (PVL genes. The staphylococcal strains were identified through partial 16S ribosomal ribonucleic acids (rRNA nucleotide sequencing, and Basic Local Alignment Search Tool (BLAST analysis of the gene sequence showed that the staphylococcal strains have 96%–100% similarity to Staphylococcus aureus (30, S. epidermidis (17, S. haemolyticus (15, S. saprophyticus (13, S. chromogenes (8, S. simulans (7, S. pseudintermedius (6 and S. xylosus (4. Resistance of 100% was observed in all Staphylococcus spp. against MET, PEN, CLN, CHL and SXT. Multi-drug resistant (MDR bacteria from nasal cavities and raw milk reveals 13 isolates were MDR against METR, PENR, AMXR, CLNR, CHLR, SXTR CLXR, KANR, ERYR, and VANR. Of all isolates, 100% harboured the mecA gene, while 30% of the isolates possess the PVL gene. All S. aureus harboured the PVL gene while other Staphylococcus spp. were negative for the PVL gene. The presence of methicillin-resistant Staphylococcus spp. isolates in dairy cattle is a potential public health risk and thus findings in this study can be used as a baseline for further surveillance.

  1. A study of biofilm production in clinical isolates of Staphylococci at a tertiary care hospital, Bangalore

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    Saroj Golia

    2015-02-01

    Full Text Available Background: The Biofilms are densely packed communities of microorganisms consisting of layers of cell clusters embedded in a matrix of extracellular polysaccharide called polysaccharide intercellular adhesin. This layer impedes the delivery of antibiotics to the biofilm forming microbial cells leading to emergence of drug resistance. Staphylococci are commensal bacteria on the human skin and mucous membranes. So it may be easily introduced as a contaminant during the surgical intervention. So, this study was conducted to identify the Biofilm producing strains from clinical isolates of Staphylococci. Methods: A total of 182 non-repetitive clinical strains of Staphylococci isolated from various clinical samples from Feb 2014 and #8211;Oct 2014 were included in the study. All the isolates were identified using standard microbiological procedures. All the samples were tested for biofilm production by modified Congo-red agar method and tube method. Results: Out of 182 samples that were included in the study, a total of 90 (49.45% samples showed biofilm formation of which 58 (75.32% were methicillin resistant and 32 (30.47% were methicillin sensitive. Also these strains were resistant to other antibiotics. Conclusion: Our study showed biofilm production by methicillin resistant strains which were also multidrug resistant. Treatment of methicillin resistant strains of Staphylococci is one of the most challenging task for the clinicians and the microbiologists. So they should be routinely screened for biofilm formation in order to prevent emergence and spread of multidrug resistant strains. [Int J Res Med Sci 2015; 3(2.000: 470-474

  2. Diversity of staphylococcal cassette chromosome mec structures in coagulase-negative staphylococci and relationship to drug resistance.

    Science.gov (United States)

    Garza-González, Elvira; López, Daniel; Pezina, Cesar; Muruet, Walter; Bocanegra-García, Virgilio; Muñoz, Ivan; Ramírez, Camilo; Llaca-Díaz, Jorge M

    2010-03-01

    The objective of this study was to determine the distribution of staphylococcal cassette chromosome mec (SCCmec) elements in meticillin-resistant coagulase-negative staphylococci (MR-CoNS) isolated from a tertiary-care hospital in Mexico and to examine the relationship to drug resistance. Fifty selected MR-CoNS isolates collected from catheters (n=15), blood (n=15), bone (n=9), bronchial lavage (n=2) and urine (n=2) and one isolate each from an abscess, cerebrospinal fluid, eye, pleural effusion, synovial fluid, tracheal aspirate and wound secretion were examined. Susceptibility testing was performed by the broth microdilution method. SCCmec types were determined by multiplex PCR and PFGE was carried out as described previously for Staphylococcus aureus. Among the MR-CoNS strains studied, the most frequently isolated species were Staphylococcus epidermidis (n=26) and Staphylococcus haemolyticus (n=13). Staphylococcus cohnii (n=5), Staphylococcus hominis (n=3), Staphylococcus sciuri (n=1), Staphylococcus pasteuri (n=1) and the recently described species Staphylococcus pettenkoferi (n=1) were also identified. The most frequent MR-CoNS genotype identified was SCCmec type IVa in S. epidermidis isolates, which also showed a high diversity in their PFGE patterns. A clone was found that amplified both SCCmec III and V elements in five isolates examined. The single MR S. pettenkoferi isolate harboured SCCmec type IVd and the single MR S. pasteuri isolate harboured SCCmec type I. The carriage of SCCmec type III was associated with resistance or intermediate resistance to meropenem (P IVa and the high genetic diversity among MR-CoNS strains. As far as is known, this is the first report describing the newly identified S. pettenkoferi possessing SCCmec IVd and S. pasteuri harbouring SCCmec type I. MR-CoNS harbouring SCCmec type III were found to be more resistant to meropenem.

  3. The use of nucleosides and arginine as alternative energy sources by coagulase-negative staphylococci in view of meat fermentation.

    Science.gov (United States)

    Janssens, M; Van der Mijnsbrugge, A; Sánchez Mainar, M; Balzarini, T; De Vuyst, L; Leroy, F

    2014-05-01

    The ability of coagulase-negative staphylococci (CNS) to use alternative energy sources in meat may partially explain their occurrence in fermented meats. Of 61 CNS strains tested, all metabolized adenosine and inosine in a meat simulation medium (MSM). The ability to catabolize arginine via the arginine deiminase (ADI) pathway varied between strains. All tested strains of Staphylococcus carnosus and Staphylococcus epidermidis possessed an arcA gene and showed ADI activity, whereas other species, such as Staphylococcus equorum and Staphylococcus succinus, did not. Arginine catabolic mobile elements (ACME), as in the positive control S. epidermidis ATCC 12228, were uncommon and only found in Staphylococcus xylosus 3PA6 (sausage isolate) and Staphylococcus chromogenes G222 (teat apex isolate). Monoculture experiments were performed in MSM with S. carnosus 833 and SS3-4, S. xylosus G211, and S. epidermidis ATCC 12228 and 2S7-4. At all pH values tested (5.3, 5.8, and 6.5), the strains of S. carnosus catabolized arginine faster than the strains of S. xylosus and S. epidermidis. Only at pH 6.5 could a low ADI activity be found for S. xylosus G211. Increased ADI activity occurred in the case of the ACME-positive S. epidermidis ATCC 12228, when compared to the ACME-negative S. epidermidis 2S7-4.

  4. Characterization and Technological Features of Autochthonous Coagulase-Negative Staphylococci as Potential Starters for Portuguese Dry Fermented Sausages.

    Science.gov (United States)

    Semedo-Lemsaddek, Teresa; Carvalho, Laura; Tempera, Carolina; Fernandes, Maria H; Fernandes, Maria J; Elias, Miguel; Barreto, António S; Fraqueza, Maria J

    2016-05-01

    The manufacture of dry fermented sausages is an important part of the meat industry in Southern European countries. These products are usually produced in small shops from a mixture of pork, fat, salt, and condiments and are stuffed into natural casings. Meat sausages are slowly cured through spontaneous fermentation by autochthonous microbiota present in the raw materials or introduced during manufacturing. The aim of this work was to evaluate the technological and safety features of coagulase-negative staphylococci (CNS) isolated from Portuguese dry fermented meat sausages in order to select autochthonous starters. Isolates (n = 104) obtained from 2 small manufacturers were identified as Staphylococcus xylosus, Staphylococcus equorum, Staphylococcus saprophyticus, and Staphylococcus carnosus. Genomically diverse isolates (n = 82) were selected for further analysis to determine the ability to produce enzymes (for example, nitrate-reductases, proteases, lipases) and antibiotic susceptibility. Autochthonous CNS producing a wide range of enzymes and showing low antibioresistance were selected as potential starters for future use in the production of dry fermented meat sausages.

  5. Occurrence and characteristics of methicillin-resistant and -susceptible Staphylococcus aureus and methicillin-resistant coagulase-negative staphylococci from Japanese retail ready-to-eat raw fish.

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    Hammad, Ahmed M; Watanabe, Wataru; Fujii, Tomoko; Shimamoto, Tadashi

    2012-06-01

    Staphylococci are not part of the normal fish microflora. The presence of staphylococci on fish is an indication of (a) post-harvest contamination due to poor personnel hygiene, or (b) disease in fish. The aim of this study was to determine the prevalence, molecular genetic characteristics, antibiotic resistance and virulence factors of methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MR-CoNS) isolated from 200 samples of retail ready-to-eat raw fish (sashimi) collected from the Japanese prefecture of Hiroshima. We characterized 180 staphylococcal strains. A majority of the grocery stores surveyed (92%, 23/25) contained fish contaminated with Staphylococcus species. We recovered 175 S. aureus isolates from 174 (87%, 174/200) samples, with 170 isolates of MSSA. For the MRSA and MR-CoNS, 10 isolates were obtained from 10 samples (5%, 10/200) collected from 10 shops (40%, 10/25) belonging to four supermarket chains. SCCmec typing revealed the presence of a type IV.1 SCCmec cassette in S. warneri isolates, a type II.1 SCCmec cassette in S. haemolyticus isolates and a cassette in methicillin-resistant S. aureus (MRSA) isolates that could not be typed. Molecular typing of two MRSA isolates by spa sequencing and multilocus sequence typing (MLST) identified t1767 and ST8, respectively. Antibiotic resistance genes that confer resistance to aminoglycosides, tetracyclines, β-lactams, macrolides, lincosamides and streptogramin B (MLS(B)) antibiotics were detected. Genes encoding one or more of the following virulence factors: staphylococcal enterotoxins (seb, and sed), toxic shock syndrome toxin 1 (tst), exfoliative toxin (etaA) were detected in 14.2% (25/175) of S. aureus isolates. The accessory gene regulator (agr) typing of S. aureus isolates revealed that agr type 1 was most prevalent (96.5%, 169/175) followed by type 2 (2.2%, 4/175) and type 3 (1.1%, 2/175). None of

  6. Estudo da produção de beta -lactamase e sensibilidade às drogas em linhagens de estafilococos coagulase-negativos isolados de recém-nascidos Study of production of beta-lactamase and drugs susceptibility in strains of coagulase-negative staphylococci isolated of neonates

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    Maria de Lourdes Ribeiro de Souza da Cunha

    2002-01-01

    Full Text Available Os estafilococos coagulase-negativos (ECN, embora reconhecidos como saprófitas por muito tempo, têm emergido como agentes etiológicos de uma série de infecções, sendo atualmente os principais responsáveis por sepse em UTI neonatal. Tendo em vista estas características, este estudo objetivou a identificação de estafilococos coagulase-negativos isolados de processos infecciosos em recém-nascidos, bem como a determinação da produção de beta-lactamase e sensibilidade às drogas pelas linhagens isoladas. O Staphylococcus epidermidis foi a espécie mais freqüentemente isolada (77,8%. O estudo da produção de beta-lactamase revelou esta característica na maioria das linhagens de ECN isoladas (71,8%. As linhagens de ECN mostraram, ainda, resistência múltipla aos antibióticos utilizados, com 63,2% dos isolados apresentando resistência a cinco ou mais drogas. A elevada transmissibilidade de plasmídios entre estas linhagens e o uso abusivo de drogas antimicrobianas têm-se constituído em importantes fatores na seleção de amostras multirresistentes e na transferência de genes de resistência.Although coagulase-negative staphylococci (CNS have been recognized as saprophytes for a long time, they had emerged as etiologic agents of infections. They have currently been the most frequently isolated pathogen in sepsis in neonatal intensive care unit (NICU. This study aimed the identification of CNS strains isolated from newborns' infections and to determination of beta-lactamase and drugs susceptibility. Staphylococcus epidermidis was the most frequently isolated species (77,8%. The study of the beta-lactamase production revealed this characteristic in the most of the strains of CNS isolated (71,8%. The strains isolated in this study presented multiple resistance to the antibiotics tested, with 63,2% of isolates presenting resistance to five or more drugs. The high transmissibility of plasmids among those strains and the abusive use of

  7. Occurrence and antimicrobial sensitivity in staphylococci isolated from goat, sheep and cow’s milk

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    Marcela Vyletělová

    2011-01-01

    Full Text Available The aim of this study was to compare the sensitivity to selected antibiotics in staphylococci isolated from goat (n = 60, sheep (n = 60 and cow’s milk (n = 120. The individual milk samples were inoculated onto Blood agar cultivated at 36 °C/24 h.The isolated species of staphylococci were identified using biochemical tests, namely STAPHYtest and identification program TNW pro 6.5. We examined the sensitivity of strains to the spectrum of antibiotics, as follows: vancomycin (VA, amoxicillin/clavulanic acid (AMC, penicillin (P, rifampicin (RD, oxacillin (OX, tetracycline (TE, erythromycin (E, chloramphenicol (C, clindamycin (DA, gentamicin (CN, ciprofloxacin (CIP, teicoplanin (TEC, cefoxitin (FOX and novobiocin (NOV. Altogether, 97 staphylococcal isolates were obtained; 70 from cow’s milk, 11 from goat’s milk and 16 from sheep’ milk. Staphylococcus aureus was the most frequent species in milk of all animal origin tested, was detected in 54 (45% cow’s milk, 10 (17% goat’s and 15 (25% sheep’ milk samples. S. lentus was identified only in goat’s and sheep’ milk whereas in cow’s milk there were representation of staphylococcal species as follows: S. haemolyticus (n = 7, S. chromogenes (n = 2, S. warneri (n = 2, S. xylosus (n = 2, S. epidermidis (n = 2 and unclassified staphylococci (n = 1. The results of S. aureus sensitivity are similar for all tested antibiotics and for all monitored milk: No resistance to vancomycin, rifampicin, chloramphenicol and teicoplanin was recorded in obtained S. aureus isolates whereas the resistance to ciprofloxacin was found out most often.

  8. Prevalence study of enterococus and staphylococci resistance to vancomycin isolated from urinary tract infections

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    Mohammad Kazem Sharifi Yazdi

    2013-07-01

    Full Text Available Background: The role of gram-positive cocci especially Staphylococci species in causing urinary tract infection are well known. Among the Staphylococci species Methicillin Resistance Staphylococcus aureus (MRSA is the most important. The rate of MRSA is increasing worldwide. This is alarming because the danger of these organism in public health. Therefore the aim of this study was to determine the sensitivity of gram-positive cocci, as well as MRSA to vancomycin and other antibiotics.Methods: This was a descriptive study, and were carried out on 300 patients with urinary tract infections (UTI caused by gram-positive cocci, referred to Imam Khomeini hospital during eight months. Prior to the antibiotic sensitivity testing all the isolates were identified according to the standard conventional biochemical procedure, and then the antibiotic susceptibility test were carried out according to Bauer-Kirby method. Results: Among the gram positive cocci causing UTI, the most abundant were Staphylococcus saprophyticus (37.7%, followed by Staphylococcus epidermidis (22.3% and Staphylococcus aureus (18% respectivley. The sex distribution of patients were 163 female (54.3% and 137 male (45.7% respectively, and the prevalence rate of urinary tract infections in female was (8.6% higher than male. The rate of sensitivity of isolated Staphylococci were as followed, sensitive to vancomycine (100%, Ciprofloxacin (89.2%, rifampin (87.6%, and amikacin (71.8% respectivley, but were resistant to penicillin and amoxicillin (100%. The antibiotic sensitivity rate of isolated  Streptococci was to vancomycine (85.1%, ciprofloxacin (50.7% and penicillin (79.1% respectively.Conclusion: Vancomycin is still a suitable antibiotic for the treatment of Staphyloco-ccus infections. Although 6% rate of enterococci resistance to vancomycin is alarming, and use of this antibiotic in the treatment of other gram-positive bacteria should be done with precaution.

  9. Community dynamics of coagulase-negative staphylococci during spontaneous artisan-type meat fermentations differ between smoking and moulding treatments.

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    Janssens, M; Myter, N; De Vuyst, L; Leroy, F

    2013-08-16

    Coagulase-negative staphylococci (CNS) that are naturally present in the raw meat batter of fermented sausages or that originate from the addition of a starter culture play a role in flavour development. A wide species diversity of CNS can be present in fermented meats, but it is not fully clear yet how specific process parameters select for specific CNS by affecting their community dynamics. Therefore, the influence of smoking and moulding treatments on the CNS community dynamics in spontaneously fermented, artisan-type sausages was investigated. During the fermentation stage, the meat batter was in all cases dominated by Staphylococcus saprophyticus, in addition to Lactobacillus sakei as governing lactic acid bacterium. Following fermentation, the bacterial communities were not perturbed by the smoking treatment, since both L. sakei and S. saprophyticus remained dominant throughout the ripening stage and prevailed in the end-products. Yet, when fermentation was followed by a moulding step with Penicillium nalgiovense, a shift of the CNS communities towards dominance by Staphyloccocus equorum was seen, despite a similar evolution of L. sakei. This effect was possibly due to a pH rise caused by the mould, a hypothesis which was reinforced by the finding that the isolated strain S. equorum DBX-S-17 was more sensitive to low pH than the isolated strain S. saprophyticus DFL-S-12 during growth experiments in brain heart infusion (BHI). Differences in CNS communities may affect sausage flavour, due to intraspecies variations in metabolic conversions of, for instance, amino acids. The fact that 3-methyl-butanal was only found in the moulded sausage, which was dominated by S. equorum, may be related to the finding that the isolated strain of this species was able to produce this compound in BHI medium, whereas the isolated strain of S. saprophyticus was not.

  10. Determination of clinical significance of coagulase-negative staphylococci in blood cultures.

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    Karakullukçu, Asiye; Kuşkucu, Mert Ahmet; Ergin, Sevgi; Aygün, Gökhan; Midilli, Kenan; Küçükbasmaci, Ömer

    2017-03-01

    The aim of this study was to investigate the criteria used to distinguish coagulase-negative staphylococci (CoNS) bacteremia from contamination. We evaluated 162 adult patients with CoNS-positive blood cultures (BCs). Of the 162 patients, 35 (21.6%) had at least 2 positive BCs and 127 (78.4%) had a single positive BC. According to the Laboratory-Confirmed Bloodstream Infection (LCBI) criteria, 24 (14.8%) patients with the same species of CoNS had true bacteremia, and 138 (85.2%) patients had contaminants. Despite the detection of the same CoNS species, 9 of the 24 patients had different CoNS genotypes. Using clinical assessments, only 20 patients were diagnosed with true bacteremia, 8 of them had a single positive BC. We concluded that only using the LCBI criteria or clinical evaluations of a patient were not sufficient to distinguish CoNS bacteremia from contamination. Molecular identification should also be performed as a diagnostic laboratory parameter for CoNS bacteremia.

  11. UVc-irradiation sublethal stress does not alter antibiotic susceptibility of staphylococci (MRSA, MSSA, and coagulase-negative staphylococci) to β-lactam, macrolide, and fluoroquinolone antibiotic agents.

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    Maeda, Yasunori; Goldsmith, Colin E; Coulter, Wilson A; Millar, B Cherie; Dooley, James S G; Lowery, Colm J; Loughrey, Anne; Rooney, Paul J; McDowell, David A; Matsuda, Motoo; Moore, John E

    2012-01-01

    Skin tanning, either by exposure to natural sunlight or through use of UV sunbeds, has become a popular practice in the US, where it is estimated that approximately 1 million times per day someone in the US uses UV radiation for skin tanning, equating to 30 million Americans (circa 10% of the US population) who use a tanning bed. As well as exposing the host to periods of UV radiation, such practices also expose commensal skin bacteria, including Staphylococcus aureus, to such UV radiation. Previous work has indicated that environmental stresses on bacteria may lead to an upregulation of stress responses, in an attempt for the organism to combat the applied stress and remain viable. UV light may act as an environmental stress on bacteria, and so it was the aim of this study to examine the effect of UVc light on the antibiotic susceptibility of commensal skin bacteria, to determine if UV radiation would increase the antibiotic resistance of such skin flora and thus lead to a potential skin flora with increased antibiotic resistance. Previously, it has been shown that UVc light has a greater mutational effect on bacteria compared to lower-energy UV forms, including UVa and UVb light. Therefore, we decided to employ UVc light in our study to amplify the potential for mutational events occurring in skin staphylococci organisms (n=8) including methicillin-sensitive Staphylococcus aureus (n=2), methicillin-resistant Staphylococcus aureus (n=4), and coagulase-negative staphylococci (Staphylococcus haemolyticus) (n=2) were exposed to varying degrees of sublethal radiation via UVc light, and their minimum inhibitory concentration (MIC) susceptibility was determined by broth dilution assay against three classes of commonly used antibiotics, namely β-lactams (penicillin), macrolides (erythromycin), and fluoroquinolones (ciprofloxacin). There was no significant difference between antibiotic susceptibility before UVc exposure and until maximum sublethal stress, prior to cell

  12. PREVALENCE AND ANTIBIOTIC RESISTANCE OF STAPHYLOCOCCI ISOLATED FROM KOLEA HOSPITAL (ALGERIA

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    M. N. Boukhatem

    2015-03-01

    Full Text Available The resurgence of multiple antibiotic resistances is a major public health problem, often causing therapeutic impasses. Our study was conducted at the Bacteriology Laboratory of Kolea (Tipaza, Algeria Hospital. The aim of this study was to determine the prevalence of staphylococci in clinical purulent, sepsis and urinary samples, and to test the sensitivity of the selected strains against a various antibiotic (ATB discs. The identification of Staphylococcus strains was performed by using classic biochemistry tests. Susceptibility against ATB was determined by the agar disk diffusion method. Among 1403 clinical samples obtained, only 61 was related to staphylococci infections (4.34%. Staphylococcus strains were found mainly in the purulent samples (60.65%. S.aureus was found to be the mainly isolated strains with a rate of 61.53% while the S. epidermidis strain was found predominantly in hospitalized patients (68.57%. The frequency of the overall resistance of S.aureus against penicillin G (80.6% and gentamicin (61.53% remains high. The multi-resistant Staphylococcus has become extremely widespread, hence the need for the accomplishment of a strategy to prevent the spread of this resistance.

  13. [Isolation of coagulase-positive staphylococci from cheese and ice-cream samples sold in Ankara and some biochemical properties of the isolates].

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    Baştepe, S; Köşker, O

    1981-01-01

    Coagulase-positive staphylococci were isolated from different kinds of cheese and ice-cream sold in Ankara and some biochemical properties of these isolates were determined. 55 cheese, 52 ice-cream (107 samples) were examined for the presence of coagulase-positive staphylococci. Baird Parker Medium was used and 26 samples constituting of 13 cheese and 13 ice-cream were found to be contaminated with coagulase-positive staphylococci and ratio of the contaminated samples to the total was calculated as 24.3%. Highest count was determined to be 176, 166/g in Izmir Tulum Cheese, whereas none of the other tulum cheese samples yielded this bacteria. In general, coagulase-positive staphylococci of cheese samples were higher than ice-cream samples. Among the ice-cream samples highest coagulase-positive staphylococci count was obtained in nutty ice-cream. From the 26 contaminated samples 164 coagulase-positive staphylococci were isolated. Results indicate that an important number of the coagulase forming isolates were also phosphatase positive, forming pigment and haemolysin, able to utilize mannitol. No relation could be observed between the coagulase formation and ability to produce lysin.

  14. Comparison of Two PCR Methods in determining the Methicillin-Resistant Gene in Coagulase-Negative Staphylococci

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    M. Bokaeian

    2016-09-01

    Full Text Available Aims: It is very important to detect the coagulase-negative Staphylococci, which produce the hospital infections. Being one of the most expensive and time-consuming stages before the polymerase chain reaction (PCR, DNA extraction is one of the primary stages of PCR. Then, it should be noticed that the elimination of the stage might save time and costs. The aim of this study was to compare two PCR methods including the method with the utilization of the extracted DNA with the extraction kit and the direct PCR method in the detection of the methicillin-resistant genes in the coagulase-negative Staphylococci. Materials & Methods: In the descriptive cross-sectional study, 135 Staphylococcus epidermidis and 88 Staphylococcus saprophyticus samples were studied, separated from blood, wounds, urinary catheter, and urine samples of patients hospitalized in the treatment centers of Zahedan. The direct PCR was done on the Staphylococcus saprophyticus and Staphylococcus epidermidis colonies. PCR with the extracted DNA was done for mecA and 16srDNA genes using the extraction kit, and the results were compared. Findings: In both methods, mecA and 16srDNA genes were successfully amplified in 310bp and 420bp related to Staphylococcus bacteria identifying gene and methicillin resistant gene, respectively. In addition, there were approximately the same band qualities. Conclusion: In order to save time and costs, the direct PCR method can be used to detect methicillin-resistant coagulase-negative Staphylococci.

  15. Number of positive blood cultures, biofilm formation, and adhesin genes in differentiating true coagulase-negative staphylococci bacteremia from contamination.

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    Papadimitriou-Olivgeri, I; Giormezis, N; Papadimitriou-Olivgeris, M; Zotou, A; Kolonitsiou, F; Koutsileou, K; Fligou, F; Marangos, M; Anastassiou, E D; Spiliopoulou, I

    2016-01-01

    The significance of the number of coagulase-negative staphylococci (CNS)-positive blood cultures remains obscure in regards to determining true bacteremia versus contamination. The goal of this study was to determine the predictors of real CNS bloodstream infection among intensive care unit (ICU) patients. ICU patients with at least one CNS-positive blood culture were identified from the microbiology database. Biofilm formation was tested by glass tube and microtiter plate assay. mecA gene, ica operon genes (icaA, icaB, icaD), and adhesin genes (aap, bap, atlE, fbe, fnbA) were detected by polymerase chain reaction (PCR). CNS were recovered from 120 septic episodes, 20 of which were true CNS bacteremias, whereas from the remaining 100 episodes, the isolated CNS were characterized as contaminants. The number of positive blood cultures was significantly associated with true CNS bacteremia. Nineteen true bacteremic Staphylococcus epidermidis strains were compared to 38 contaminants. Biofilm synthesis was documented in 37 isolates associated with the presence of the ica operon (p = 0.048). There were 39, 26, 38, 21, and 10 strains positive for the presence of atlE, bap, fbe, aap, and fnbA genes, respectively. Rifampicin resistance, absence of severe sepsis, number of S. epidermidis-positive blood cultures, and absence of the bap gene were independently associated with true S. epidermidis bacteremia as compared to contaminant strains. The number of positive blood cultures is associated with true CNS bacteremia. The presence of adhesin genes may play a role in differentiating true infection from contamination, whereas absence of the bap gene is associated with true S. epidermidis bacteremia.

  16. Trends in antibiotic resistance in coagulase-negative staphylococci in the United States, 1999 to 2012.

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    May, Larissa; Klein, Eili Y; Rothman, Richard E; Laxminarayan, Ramanan

    2014-01-01

    Coagulase-negative staphylococci (CoNS) are important bloodstream pathogens that are typically resistant to multiple antibiotics. Despite the concern about increasing resistance, there have been no recent studies describing the national prevalence of CoNS pathogens. We used national resistance data over a period of 13 years (1999 to 2012) from The Surveillance Network (TSN) to determine the prevalence of and assess the trends in resistance for Staphylococcus epidermidis, the most common CoNS pathogen, and all other CoNS pathogens. Over the course of the study period, S. epidermidis resistance to ciprofloxacin and clindamycin increased steadily from 58.3% to 68.4% and from 43.4% to 48.5%, respectively. Resistance to levofloxacin increased rapidly from 57.1% in 1999 to a high of 78.6% in 2005, followed by a decrease to 68.1% in 2012. Multidrug resistance for CoNS followed a similar pattern, and this rise and small decline in resistance were found to be strongly correlated with levofloxacin prescribing patterns. The resistance patterns were similar for the aggregate of CoNS pathogens. The results from our study demonstrate that the antibiotic resistance in CoNS pathogens has increased significantly over the past 13 years. These results are important, as CoNS can serve as sentinels for monitoring resistance, and they play a role as reservoirs of resistance genes that can be transmitted to other pathogens. The link between the levofloxacin prescription rate and resistance levels suggests a critical role for reducing the inappropriate use of fluoroquinolones and other broad-spectrum antibiotics in health care settings and in the community to help curb the reservoir of resistance in these colonizing pathogens.

  17. Genetic characterization of coagulase-positive staphylococci isolated from healthy pigeons.

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    Kizerwetter-Świda, M; Chrobak-Chmiel, D; Rzewuska, M; Antosiewicz, A; Dolka, B; Ledwoń, A; Czujkowska, A; Binek, M

    2015-01-01

    Coagulase-positive staphylococci (CoPS) are opportunistic veterinary pathogens, of which Staphylococcus aureus, S. delphini and S. intermedius can be isolated from pigeons. The biochemical identification of S. delphini and S. intermedius isolates may be incorrect, because of their phenotypic similarity. The purpose of the present study was to isolate and identify CoPS from domestic and feral pigeons and to determine their genetic relatedness by PFGE. A total number of 31 isolates of CoPS were obtained, 15 were identified as S. delphini group B, six as S. aureus, four as S. delphini group A, three as S. intermedius and three as S. schleiferi subsp. coagulans. The results indicate that S. delphini group B is the predominant CoPS species among pigeons studied. PFGE restriction patterns of S. delphini group A and S. delphini group B form separate clusters, demonstrating their genetic heterogeneity. Indistinguishable or very similar PFGE patterns observed among S. delphini group B isolates from domestic and feral pigeons confirm the possibility of CoPS transmission between these birds.

  18. Acceleration of the direct identification of Staphylococcus aureus versus coagulase-negative staphylococci from blood culture material: a comparison of six bacterial DNA extraction methods.

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    Loonen, A J M; Jansz, A R; Kreeftenberg, H; Bruggeman, C A; Wolffs, P F G; van den Brule, A J C

    2011-03-01

    To accelerate differentiation between Staphylococcus aureus and coagulase-negative staphylococci (CNS), this study aimed to compare six different DNA extraction methods from two commonly used blood culture materials, i.e. BACTEC and BacT/ALERT. Furthermore, we analysed the effect of reduced blood culture incubation for the detection of staphylococci directly from blood culture material. A real-time polymerase chain reaction (PCR) duplex assay was used to compare the six different DNA isolation protocols on two different blood culture systems. Negative blood culture material was spiked with methicillin-resistant S. aureus (MRSA). Bacterial DNA was isolated with automated extractor easyMAG (three protocols), automated extractor MagNA Pure LC (LC Microbiology Kit M(Grade)), a manual kit MolYsis Plus and a combination of MolYsis Plus and the easyMAG. The most optimal isolation method was used to evaluate reduced bacterial incubation times. Bacterial DNA isolation with the MolYsis Plus kit in combination with the specific B protocol on the easyMAG resulted in the most sensitive detection of S. aureus, with a detection limit of 10 CFU/ml, in BacT/ALERT material, whereas using BACTEC resulted in a detection limit of 100 CFU/ml. An initial S. aureus or CNS load of 1 CFU/ml blood can be detected after 5 h of incubation in BacT/ALERT 3D by combining the sensitive isolation method and the tuf LightCycler assay.

  19. Coagulase-negative staphylococci causing blood stream infection at an Indian tertiary care hospital: Prevalence, antimicrobial resistance and molecular characterisation

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    S Singh

    2016-01-01

    Full Text Available Introduction: Recent years have seen a rise of coagulase-negative staphylococci (CoNS from common contaminants to agents of nosocomial blood stream infections (BSI's. Molecular typing and establishing a correlation with antibiotic resistance is essential particularly in countries like India where genotyping studies for drug-resistant CoNS are sparse. Methods: A prospective study was done over 18 months, wherein 42,693 blood samples were received, and 59 patients with BSI due to CoNS were evaluated. The isolates recovered were identified by a biochemical test panel and matrix-assisted laser desorption ionization – time of flight mass spectrometry followed by antimicrobial susceptibility testing by Kirby–Baur disc diffusion method and E-test strips. Staphylococcal chromosomal cassette mec (SCCmec element was characterised by multiplex polymerase chain reaction for all methicillin-resistant (MR isolates. Results: The majority of CoNS isolated were constituted by Staphylococcus haemolyticus (47.5% followed by Staphylococcus epidermidis (33.9%, Staphylococcus hominis (11.86%, Staphylococcus cohnii (5.08% and Staphylococcus warneri (1.69%. Among all isolates 57.6% were MR with statistically significant higher resistance versus methicillin sensitive-CoNS. This difference was significant for erythromycin (76% vs. 44%, P = 0.011, rifampicin (50% vs. 12%,P= 0.002 and amikacin (26.5% vs. 4%, P = 0.023, ciprofloxacin (64.7% vs. 20%, P = 0.001 and cotrimoxazole (55.9% vs. 20%, P = 0.006. SCCmec type I was predominant (61.8%, P = 0.028 and exhibited multidrug resistance (76.2%. Coexistence of SCCmec type I and III was seen in 8.82% MR isolates. Conclusion: CoNS exhibit high antimicrobial resistance thereby limiting treatment options. The presence of new variants of SCCmec type in hospital-acquired CoNS may predict the antibiotic resistance pattern. This is the first evaluation of the molecular epidemiology of CoNS causing BSI from India and can serve as a

  20. [Investigation of biofilm-associated antibiotic susceptibilities of methicillin-resistant staphylococci isolated from catheter-related nosocomial infections].

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    Bayındır Bilman, Fulya; Can, Füsun; Kaya, Melek; Yazıcı, Ayşe Canan

    2013-07-01

    Risks for development of local and/or systemic infections are the most important complications of catheters that are widely used during hospitalization process. The aims of this study were to investigate and compare the antibiotic susceptibilities of methicillin-resistant staphylococci isolated from catheters, in planktonic and biofilm forms, and to evaluate the antimicrobial effects of antibiotics on those forms alone and in combinations. A total of 30 strains [15 methicillin-resistant Staphylococcus aureus (MRSA) and 15 methicillin-resistant coagulase-negative staphylococci (MR-CNS)] isolated from catheter cultures of patients hospitalized in different clinics and intensive care units in Baskent University Medical School Hospital between 2006-2009, were included in the study. The antibiotic sensitivities of MRSA and MR-CNS isolates were investigated in vitro in planktonic phase and on sessile cells after biofilm was formed. Vancomycin, ciprofloxacin, rifampicin, gentamicin, meropenem, tigecycline, linezolid, ceftazidime and cephazolin were used for antibiotic susceptibility testing. The sensitivity of planktonic cells to antibiotics was primarily investigated, so that minimal inhibitor concentration (MIC) and minimal bactericidal concentration (MBC) values were determined by broth microdilution method. Afterwards, each strain was transformed to sessile cell in a biofilm environment, and MIC and MBC values were also determined for sessile cells. Double and triple antibiotic combinations were prepared, the effectiveness of combinations were studied on both planktonic and biofilm cells with multiple-combination bactericidal testing (MCBT) method. The data set obtained from planktonic and biofilm cells for each antibiotic analyzed via two proportion z test. Statistically significant decreases were found in the sensitivities of sessile cells when compared to planktonic cells (pantibiotic combinations also showed the susceptibility decrease between planktonic and

  1. Intramammary infections and teat canal colonization with coagulase-negative staphylococci after postmilking teat disinfection: species-specific responses.

    Science.gov (United States)

    Quirk, T; Fox, L K; Hancock, D D; Capper, J; Wenz, J; Park, J

    2012-04-01

    Coagulase-negative staphylococci (CNS) are the most common pathogens associated with intramammary infections (IMI) in dairy cows. We hypothesized that postmilking teat disinfection would reduce microbial colonization of the teat canal and thus reduce the prevalence of IMI caused by certain CNS species. The efficacy of iodine postmilking teat dip was tested against CNS colonization of the teat canal, and incidence of IMI was measured. Using an udder-half model, 43 Holstein cows at the Washington State University Dairy were enrolled in the trial; postmilking teat dip was applied to one udder-half, treatment (TX), and the remaining half was an undipped control (CX). Teat canal swabbing and mammary quarter milk samples were taken in duplicate once a week for 16 wk for microbial culture. Isolates from agar cultures were presumptively identified as CNS and then speciated using PCR-RFLP and agarose gel electrophoresis. Colonization of the teat canal and IMI by CNS were assessed. Thirty CNS IMI were diagnosed and the number of new IMI in CX quarters (21) was significantly greater than that in TX mammary quarters (9). The majority of CNS IMI were caused by Staphylococcus chromogenes (30%) and Staphylococcus xylosus (40%), and the latter were appreciably reduced by teat dip. Except for S. xylosus, an association was observed between teat canal colonization and IMI by all CNS species in this study, in which the majority of IMI were preceded by teat canal colonization. The total number of CNS IMI was greater for CX group cows compared with TX group cows. However, the effect of disinfection on IMI did not appear to be the same for all CNS species.

  2. 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.

  3. Coagulase-negative staphylococci as reservoirs of genes facilitating MRSA infection

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    Otto, Michael

    2013-01-01

    Recent research has suggested that Staphylococcus epidermidis is a reservoir of genes that, after horizontal transfer, facilitate the potential of Staphylococcus aureus to colonize, survive during infection, or resist antibiotic treatment, traits that are notably manifest in methicillin-resistant S. aureus (MRSA). S. aureus is a dangerous human pathogen and notorious for acquiring antibiotic resistance. MRSA in particular is one of the most frequent causes of morbidity and death in hospitalized patients. S. aureus is an extremely versatile pathogen with a multitude of mechanisms to cause disease and circumvent immune defenses. In contrast, most other staphylococci, such as S. epidermidis, are commonly benign commensals and only occasionally cause disease. Recent findings highlight the key importance of efforts to better understand how genes of staphylococci other than S. aureus contribute to survival in the human host, how they are transferred to S. aureus, and why this exchange appears to be uni-directional. PMID:23165978

  4. Peptide Nucleic Acid-Fluorescence In Situ Hybridization for Detection of Staphylococci From Endophthalmitis Isolates: A Proof-of-Concept Study.

    Science.gov (United States)

    Patel, Nimesh; Miller, Darlene; Relhan, Nidhi; Flynn, Harry W

    2017-08-01

    Rapid identification of pathogens causing endophthalmitis may improve treatment outcomes through early administration of species-specific medication. The current study reports a new molecular application of peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) with Staphylococcus-specific molecular PNA probes for the potential rapid detection of common pathogens causing endophthalmitis. An experimental study was designed to evaluate the proof of concept at the microbiology laboratory of the Bascom Palmer Eye Institute. Stored culture-positive staphylococci endophthalmitis isolates obtained from prior vitreous samples (n = 15), along with broth as negative controls (n = 5) were used. Inoculum was prepared to a final concentration of 1 × 105 colony-forming units/mL to ensure that the isolates were viable. Smears of samples were fixed and hybridized using QuickFISH protocol with probes for Staphylococcus. With PNA-FISH technique, Staphylococcus aureus was identified in 9 of 10 samples and coagulase-negative staphylococci were identified in 10 of 10 samples. Detection time was 20 minutes. This study serves a proof of concept using a new microbial detection system with FISH probes, and may have the potential for clinical use in the rapid and accurate identification of isolates from patients with endophthalmitis.

  5. Different susceptibility of coagulase-positive and coagulase-negative staphylococci to ciprofloxacin.

    Science.gov (United States)

    Utili, R; Tripodi, M F; Rosario, P; Andreana, A; Locatelli, A; Rambaldi, A; Florio, A

    1996-10-01

    Ciprofloxacin susceptibility was evaluated in 573 clinical staphylococcal isolates (Staphylococcus aureus 230, methicillin-resistant (MR) 36%; coagulase-negative strains 343, MR 66%) collected from 1989 to 1995. Resistance to ciprofloxacin for MR Staphylococcus aureus was 25% until 1991 when ciprofloxacin was introduced into the hospital formulary, and rose progressively to 90% in 1994-1995. MR Staphylococcus haemolyticus showed a ciprofloxacin resistance of 46% until 1991 and of 73% from 1992 to 1995. In contrast MR Staphylococcus epidermidis and other coagulase-negative strains showed a constant susceptibility to this agent (80%). Ciprofloxacin has limited usefulness against MR Staphylococcus aureus but can be still used to treat Staphylococcus epidermidis infections.

  6. Antimicrobial susceptibility pattern and SCCmec types of methicillin-resistant coagulase-negative staphylococci from subclinical bovine mastitis in Hatay, Turkey

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    Aslantaş Özkan

    2014-12-01

    Full Text Available Eighty-nine isolates of coagulase-negative staphylococci (CoNS of eight species from subclinical bovine mastitis were screened for the phenotypic and genotypic methicilline-resistance. In addition, all methicillin-resistant (MR isolates indicating the mecA gene were examined by PCR for the antimicrobial susceptibility patterns, and staphylococcal cassette chromosome mec (SCCmec types were also determined by multiplex PCR. A total of 21 (23.6% CoNS isolates were found to be resistant to oxacillin in broth microdilution assay. All isolates phenotypically resistant to oxacillin did not have the mecA gene, which was only found in 14.6% (13 of the isolates. Most MR-CoNS isolates were highly resistant to erythromycin (92.3%, fusidic acid (84.6%, penicillin (76.9%, and rifampycin (61.5%, and susceptible to mupirocin (100%, tetracycline (100%, vancomycin (100%, clindamycin (92.3%, and sulfamethoxazole-trimethoprim (69.2%. In conclusion, a high rate of antimicrobial resistance among MR-CoNS isolated from food producing animals emphasises the need for periodic surveillance of their resistance.

  7. Implementation of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry in Routine Clinical Laboratories Improves Identification of Coagulase-Negative Staphylococci and Reveals the Pathogenic Role of Staphylococcus lugdunensis.

    Science.gov (United States)

    Argemi, Xavier; Riegel, Philippe; Lavigne, Thierry; Lefebvre, Nicolas; Grandpré, Nicolas; Hansmann, Yves; Jaulhac, Benoit; Prévost, Gilles; Schramm, Frédéric

    2015-07-01

    The use of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for staphylococcal identification is now considered routine in laboratories compared with the conventional phenotypical methods previously used. We verified its microbiological relevance for identifying the main species of coagulase-negative staphylococci (CoNS) by randomly selecting 50 isolates. From 1 January 2007 to 31 August 2008, 12,479 staphylococci were isolated with phenotypic methods, of which 4,594 were identified as Staphylococcus aureus and 7,885 were coagulase negative staphylococci. Using MALDI-TOF MS from 1 January 2011 to 31 August 2012, 14,913 staphylococci were identified, with 5,066 as S. aureus and 9,847 as CoNS. MALDI-TOF MS allowed the identification of approximately 85% of the CoNS strains, whereas only 14% of the CoNS strains were identified to the species level with phenotypic methods because they were often considered contaminants. Furthermore, the use of MALDI-TOF MS revealed the occurrence of recently characterized Staphylococcus species, such as S. pettenkoferi, S. condimenti, and S. piscifermentans. Microbiological relevance analysis further revealed that some species displayed a high rate of microbiological significance, i.e., 40% of the S. lugdunensis strains included in the analysis were associated with infection risk. This retrospective microbiological study confirms the role of MALDI-TOF MS in clinical settings for the identification of staphylococci with clinical consequences. The species distribution reveals the occurrence of the recently identified species S. pettenkoferi and putative virulent species, including S. lugdunensis. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  8. Identification of Variable Traits among the Methicillin Resistant and Sensitive Coagulase Negative Staphylococci in Milk Samples from Mastitic Cows in India

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    Sudipta Mahato

    2017-07-01

    Full Text Available Methicillin resistant Staphylococcus aureus causing bovine mastitis has been very well investigated worldwide. However, there are only limited reports on the characterization of methicillin resistant and sensitive coagulase negative staphylococci (CoNS across the globe. Hence, in the present study, we aim to determine the phenotypic traits based on antimicrobial susceptibility profile and genotypic characterization by verifying the presence of resistance determinants, virulence and toxin genes present in the CoNS causing clinical mastitis. We obtained 62 CoNS isolates from 167 mastitic milk samples collected from three different states of India. The 62 isolates comprises of 10 different CoNS species S. sciuri, S. haemolyticus, S. chromogenes, S. saprophyticus, S. xylosus, S. simulans, S. agnetis, S. epidermidis, S. gallinarum, and S. cohinii. Susceptibility screening against 11 antibiotics determined 45.16% isolates as multidrug resistant (resistant to more than two class of antibiotic, 46.74% resistant (one or two antibiotic class and 8.06% isolates were pan-sensitive (sensitive to all drugs. High resistance was observed against oxacillin and cefoxitin, whereas all isolates were susceptible toward vancomycin and linezolid. Fifty three isolates were methicillin resistant and 9 isolates were sensitive as determined by oxacillin susceptibility assay. The methicillin resistance gene, mecA was found in 95.16% isolates and staphylococcal cassette chromosome mec (SCCmec typing predominantly revealed Type III (n = 34 and Type V (n = 18. Interestingly, 11.9% of mecA positive isolates were oxacillin susceptible and referred as oxacillin susceptible mecA positive staphylococci (OS-MRS. Additionally, genes encoding for enterotoxin, (sea, seb, seh, see toxic shock syndrome (tsst, exfoliatin (eta, etb, etd and virulence (pvl, Y-hlg were also screened. Of all the genes examined, 67.74% of isolate were positive for the Y-hlg gene, followed by the sea gene in

  9. Species identification, slime production and oxacillin susceptibility in coagulase-negative staphylococci isolated from nosocomial specimens Identificação de espécies, produção de "slime" e sensibilidade a oxacilina em amostras de Staphylococcus coagulase-negativo isoladas de espécimes nosocomiais

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    Lucía E. Alcaráz

    2003-04-01

    Full Text Available Ninety-two coagulase negative staphylococci (CNS (forty-five of clinical origin and forty-seven of environmental origin, collected in a hospital in San Luis, Argentina, from March to June, 1999, were identified to species level by the ID 32 Staph and API Staph System (bioMérieux. Slime production was investigated by the quantitative and qualitative methods. Oxacillin susceptibility was determined by the disk diffusion test (1 µg, the agar dilution method (0.125 to 4 mg/ml and agar screen (6 µg/ml. The presence of mecA gene was investigated by PCR. The clinical CNS species most commonly isolated were S. epidermidis, S. haemolyticus, S. hominis and S. saprophyticus. The frequency of slime production by clinical and environmental isolates was similar (25/45 and 27/47, respectively and the results obtained by the quantitative and the qualitative methods correlated well. The mecA gene was detected in all S. epidermidis, S. haemolyticus and S. hominis isolates, which were resistant to oxacillin by the phenotypic methods. However, this gene was not present in S. klossii, S. equorum, S. xylosus and S. capitis strains. The gene was neither found in two out of the six S. saprophyticus isolates, in two out of three S. cohnii subsp. urealyticum isolates and in two out of five S. cohnii subsp. cohnii isolates, all of which resulted oxacillin resistant according to MIC. The gene was not found in oxacillin-susceptible strains either. Most of the CNS isolates (enviromental and clinical that were slime producers were found to be oxacillin resistant, which makes the early detection of these microorganisms necessary to prevent their dissemination in hospitals, particularly among immunocompromised patients.Noventa e duas amostras de Staphylococcus coagulase negativo (SCN, (45 amostras clínicas e 47 ambientais, coletadas em um hospital de San Luis, Argentina, durante o período de março a junho de 1999, foram identificadas até espécies, empregando-se os

  10. Antibiotic susceptibility of coagulase-negative staphylococci (CoNS): emergence of teicoplanin-non-susceptible CoNS strains with inducible resistance to vancomycin.

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    Ma, Xiao Xue; Wang, En Hua; Liu, Yong; Luo, En Jie

    2011-11-01

    Coagulase-negative staphylococci (CoNS) have become increasingly recognized as important agents of nosocomial infection. One of the characteristics of CoNS is their resistance to multiple antimicrobial agents commonly used for the treatment of staphylococcal infections. CoNS strains (n = 745) isolated from a university teaching hospital in China between 2004 and 2009 were tested for antibiotic resistance. The antibiotics were placed into three categories based on resistance levels of the CoNS strains to these antibiotics: high resistance (resistance rate >70 %), including penicillin G, oxacillin and erythromycin; medium resistance (resistance rate between 30 and 70 %), including tetracycline, clindamycin, ciprofloxacin, trimethoprim/sulfamethoxazole and chloramphenicol; and low resistance (resistance rate CoNS is needed to determine their resistance to glycopeptides.

  11. First identification of methicillin-resistant Staphylococcus pseudintermedius strains among coagulase-positive staphylococci isolated from dogs with otitis externa in Trinidad, West Indies

    OpenAIRE

    2015-01-01

    Background: Otitis externa is a common inflammatory ear disease in dogs caused by a variety of pathogens, and coagulase-positive staphylococci are frequently isolated from such infections.Objective: To identify antimicrobial susceptibility profiles and methicillin-resistant strains among coagulase-positive staphylococci isolated from otitis externa in dogs.Methods: A cross-sectional study was performed over 2 years on 114 client-owned dogs presented to the Veterinary Teaching Hospital with a ...

  12. 江苏地区奶牛乳房炎凝固酶阴性葡萄球菌耐药基因分析%Analysis of antibiotic resistance and resistance genes in coagulase-negative Staphylococci (CNS) isolated from bovine mastitis in Jiangsu area

    Institute of Scientific and Technical Information of China (English)

    林伟东; 宰栩生; 于恩琪; 秦爱建; 金文杰; 梁祥焕; 王锦荣; 张军

    2015-01-01

    对江苏地区多个奶牛场采样,并对奶牛乳房炎(Bovine mastitis,BMT)检测阳性奶样进行病原菌分离、鉴定.对分离出的凝固酶阴性葡萄球菌(Coagulase-negative Staphylococci,CNS)分别进行20种药物的敏感性试验和相关耐药基因检测.结果显示,BMT检测阳性的奶样有610份,528份检出细菌,共检出细菌802株,鉴定为CNS的217株;大部分CNS对万古霉素、左氟沙星、氯霉素、磷霉素、呋喃妥因敏感,对青霉素、大观霉素具有较强耐药性;CNS对β-内酰胺类(blaZ)、大环内酯类(ermC)和四环素类(tetK、tetM、tetL、tetO)耐药基因的检出率分别为29.95%、2.30%、24.88%、1.84%、1.84%和0.00%.

  13. Frequency and Antimicrobial Susceptibility of Coagulasenegative Staphylococci Isolated from Bovine Mastitis in Dairy Farms from Tolima, Colombia

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    María del Pilar Sánchez Bonilla

    2015-09-01

    Full Text Available Objective: To determine the antimicrobial presence and susceptibility of the coagulase-negative staphylococci group (ECN, for its initials in Spanish in some cattle farms from Tolima, Colombia. Materials and methods: Using the California test for the diagnosis of mastitis (CMT, for its initials in Spanish, 484 quarters belonging to 121 cows from five small production ranches from a region of central Tolima were evaluated. CMT positive samples were cultivated for bacterial isolation. The ECN found were tested for susceptibility to the antibiotics. The results were analyzed with descriptive statistics. Results: 252 (52% quarters did not show any type of reaction to the CMT, nor did they show any clinical signs of mastitis, therefore they were considered free of the disease. From the quarters, 73 (15% turned positive for CMT and bacteriological culture. From these, 36 strains of ECN were isolated (7.4% of the total of quarters; S. aureus-ECP, 28 (5.8%; Streptococcus spp., 6 (1.2%.; Escherichia coli, 2 (0.4%, and Klebsiella pneumoniae, 2 (0.2%. The clinical and subclinical mastitis in the quarter occurred in 1.4% and 13.6%, respectively. In 5 (1.0% of the quarters, clinical mastitis caused by ECN was found and subclinical mastitis in 31 (6.4%. 61% of the ECN strains were resistant to penicillin, and 58%, to tetracycline; 97% were sensitive to cefoperazone. Conclusion: The ECN group, considered a global emergent of mastitis, is evidenced with high frequency in ranches from Tolima, Colombia, causing clinical and subclinical mastitis with varied response to antimicrobials.

  14. Oxacilin-resistant Coagulase-negative staphylococci (CoNS) bacteremia in a general hospital at São Paulo city, Brasil.

    Science.gov (United States)

    d'Azevedo, P A; Secchi, C; Antunes, A L S; Sales, T; Silva, F M; Tranchesi, R; Pignatari, A C C

    2008-10-01

    In the last decades, coagulase-negative staphylococci (CoNS), especially Staphylococcus epidermidis have become an important cause of bloodstream infections. In addition, rates of methicillin-resistance among CoNS have increased substantially, leading to the use of glicopeptides for therapy. The objective of this study was to evaluate eleven consecutives clinically relevant cases of oxacillin-resistant CoNS bacteremia in a general hospital localized in São Paulo city, Brazil. Five different species were identified by different phenotypic methods, including S. epidermidis (5), S. haemolyticus (3), S. hominis (1), S. warneri (1) and S. cohnii subsp urealyticus (1). A variety of Pulsed Field Gel Electrophoresis profiles was observed by macrorestriction DNA analysis in S. epidermidis isolates, but two of three S. haemolyticus isolates presented the same profile. These data indicated the heterogeneity of the CoNS isolates, suggesting that horizontal dissemination of these microorganisms in the investigated hospital was not frequent. One S. epidermidis and one S. haemolyticus isolates were resistant to teicoplanin and susceptible to vancomycin. The selective pressure due to the use of teicoplanin in this hospital is relevant.

  15. The evaluation of staphylococci strains isolated from nasal and bone cultures in patients with chronic osteomyelitis

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    Mehmet Uluğ

    2012-09-01

    Full Text Available Objectives: In this study, it was aimed to determine theprevalence of nasal carriage of S. aureus in chronic osteomyelitis,assessed trends in methicillin resistance withtime, and evaluated the bone and nasal cultures in staphylococcalchronic osteomyelitis.Materials and methods: Bone and nasal cultures wereperformed intra-operative and pre-operatively from 43patients with chronic osteomyelitis. Inoculation was performedon 5% sheep blood and eosine-methylen-blueagar. The samples were incubated at 37°C for 24 hours.Catalase and coagulase tests were performed on Grampositive coccus strains. Antimicrobial susceptibilities of allS. aureus strains were evaluated by disc diffusion methodaccording to CLSI for oxacillin and other antibiotics.Results: In this study pre-operative nasal cultures andintra-operative bone cultures obtained between May 2005and September 2006 were evaluated retrospectively. Ofthe 43 nasal cultures, 31 (72% yielded staphylococcalstrain, of these 18 (58% were S. aureus. Of the 18 S.aureus strains, 13 (72.2% have methicilline resistance.On the other hand, of the 43 bone cultures, 29 (67.4%yielded staphylococci strain, of these 23 (79.3% were S.aureus. Of the 23 S. aureus strains, 52.1% have methicillineresistance. Overall the similarity rate of staphylococcalstrains was 38% (11/29, while considering the statusof strains resistant to antibiotics; this ratio was 24% (7/29.Conclusions: Nasal S. aureus carriage rate (58% in patientswith chronic osteomyelitis was higher than the communityand the other patient groups. However, the surveillancecultures can give knowledge about the causativepathogen of 25% of the culture negative cases.Key words: Chronic osteomyelitis, nasal carriage, Staphylococcusaureus, bone culture

  16. Phenotypic and technological diversity of lactic acid bacteria and staphylococci isolated from traditionally fermented sausages in southern Greece.

    Science.gov (United States)

    Drosinos, Eleftherios H; Paramithiotis, Spiros; Kolovos, George; Tsikouras, Ioannis; Metaxopoulos, Ioannis

    2007-05-01

    The physicochemical and microbiological characteristics of spontaneously fermented sausages made by two medium-sized enterprises (MSE) located in southern Greece have been studied. A total of 300 lactic acid bacteria and 300 staphylococcal strains have been isolated and identified by their physiological characteristics. Lactobacillus plantarum strains were found to dominate the lactic acid bacteria microbiota in most of the cases with L. sakei strains prevailing in some of them and L. rhamnosus strains occasionally accompanying the dominant lactic acid bacteria microbiota. On the other hand, S. saprophyticus strains were found to dominate the staphylococcal microbiota in all spontaneously fermented sausages with of S. simulans, S. xylosus, S. gallinarum and S. cohnii cohnii strains being sporadically present. Following the identification, an evaluation of their technological properties, namely proteolytic and lipolytic capacities as well as production of biogenic amines and antimicrobial compounds, took place. None of the lactic acid bacteria and staphylococci was found to possess lipolytic activity whereas a total of 6 lactic acid bacteria and 51 staphylococci strains were found to be able to hydrolyse either the sarcoplasic, myofibrillar or both protein fractions. Furthermore, only one L. sakei strain and 185 staphylococci strains were found to possess decarboxylase activity against lysine, tyrosine, ornithine or histidine. Finally none of the staphylococcal microbiota and 3 lactic acid bacteria strains were found to be able to produce antimicrobial compounds of proteinaceous nature against Listeria monocytogenes.

  17. mecA-related structure in methicillin-resistant coagulase-negative staphylococci from street food in Taiwan

    Science.gov (United States)

    Yang, Tsung-Ying; Hung, Wei-Wen; Lin, Lin; Hung, Wei-Chun; Tseng, Sung-Pin

    2017-01-01

    Antibiotic-resistant patterns, a mecA homologue complex, and staphylococcal cassette chromosome mec (SCCmec) were analysed in samples of ready-to-eat (RTE) street food in Taiwan. RTE food samples (270) were collected in three densely populated Taiwanese cities between June and November 2014. Among 14 strains being identified as methicillin-resistant coagulase-negative staphylococci (MRCoNS), genetic diversities was determined by PFGE analysis. SCCmec types IV, V, VIII and TXG-24 were detected in 9, and mecASs (a mecA homologue) detected in 8. The mecASs gene complex from S. sciuri subsp. sciuri TXG-24 was found to be closely related to those found in both S. sciuri subsp. sciuri (ATCC29062) and S. sciuri subsp. rodentium (ATCC700061). SCCmecTXG24 carries a class A mec complex, a ccrA5B3-like gene complex, a heavy metal gene complex, and an IS1216 mobile element carrying tet(S). Matching identity to ccrA5 was 84.5% for ccrA in S. pseudintermedius KM241. Matching identify to ccrB3 was 92.1% for ccrB in S. pseudintermedius AI16. Similar ccrA and SCCmec boundary sequences suggest that SCCmec is easily transmitted to coagulase-negative staphylococci (CoNS). Based on MRCoNS strains identified in this research, Taiwanese RTE food products likely carry multiple antibiotic resistance genes that can be transmitted to hospitals and other clinical settings. PMID:28181543

  18. Evaluation of oxacillin and cefoxitin disks for detection of resistance in coagulase negative staphylococci

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    Ana Lúcia Souza Antunes

    2007-09-01

    Full Text Available Coagulase-negative Staphylococcus spp. was considered nonpathogenic until the emergence of multiresistance and the demonstration of their participation as infectious agents. In Brazil, oxacillin resistance may be present in over 80% of isolates, and the Clinical and Laboratory Standards Institute standardized a disk-diffusion method to predict this resistance in Staphylococcus. The aim of this study was to evaluate the variability among commercial disks of oxacillin (1 µg and cefoxitin (30 µg widely used in clinical laboratories of microbiology, compared with mecA gene and minimum inhibitory concentration (MIC of oxacillin. The use of oxacillin and cefoxitin disks simultaneously allowed the detection of important differences, particularly, in less frequent species such as S. cohnii, S. haemolyticus, S. saprophyticus, and S. sciuri. Disks of cefoxitin of the brand 2 displayed good correlation with the mecA gene (98.7% and oxacillin MIC (97.8%, while major discrepancies were observed using disks of brand 1. One of the critical points in the diffusion disk test is the quality of the disks: the use of better quality disks associated with molecular methods lead to better results to define the best antibiotic therapy.

  19. The relationship between the use of flucloxacillin, vancomycin, aminoglycosides and ciprofloxacin and the susceptibility patterns of coagulase-negative staphylococci recovered from blood cultures.

    NARCIS (Netherlands)

    Mulder, JG; Kosterink, JGW; Degener, JE

    1997-01-01

    Antibiotic use is a cause of selection of multiresistant bacterial strains. Over three years (1990-1992) we studied the relation between the use of flucloxacillin, vancomycin, aminoglycosides and ciprofloxacin and the susceptibility of coagulase-negative staphylococci (CNS) recovered from blood

  20. Acceleration of direct identification of S.aureus versus Coagulase Negative Staphylococci from blood culture material: a comparison of six bacterial DNA extraction methods

    NARCIS (Netherlands)

    Drs H. Kreeftenberg; Dr. Ir. P.F.G. Wolffs; Drs A.R. Jansz; Prof. Dr. C.A. Bruggeman; Drs A.J.M. Loonen; Dr. A.J.C. van den Brule, van den

    2010-01-01

    To accelerate differentiation between Staphylococcus aureus and Coagulase Negative Staphylococci (CNS), this study aimed to compare six different DNA extraction methods from 2 commonly used blood culture materials, i.e. BACTEC and Bact/ALERT. Furthermore, we analyzed the effect of reduced blood

  1. Acceleration of direct identification of S.aureus versus Coagulase Negative Staphylococci from blood culture material: a comparison of six bacterial DNA extraction methods

    NARCIS (Netherlands)

    Loonen, A.J.M.; Jansz, A.R.; Kreeftenberg, H.; Bruggeman, C.A.; Wolffs, P.F.G.; Brule, van den A.J.C.

    2010-01-01

    To accelerate differentiation between Staphylococcus aureus and Coagulase Negative Staphylococci (CNS), this study aimed to compare six different DNA extraction methods from 2 commonly used blood culture materials, i.e. BACTEC and Bact/ALERT. Furthermore, we analyzed the effect of reduced blood cult

  2. In vitro antimicrobial susceptibility of staphylococci isolated from canine pyoderma in Rio de Janeiro, Brazil Susceptibilidade in vitro a antimicrobianos de staphylococci isolados de cães com piodermatite no estado do Rio de Janeiro, Brasil

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    B. Penna

    2009-09-01

    Full Text Available The study aimed to determine the etiology and antimicrobial resistance of staphylococci isolated from canine pyoderma. Samples were obtained from dogs with pyoderma and isolates were identified by biochemical reactions and tested for susceptibility to 15 antimicrobials. Thirty nine staphylococci isolates were obtained, and S. pseudintermedius was the most frequent (47.4%. All isolates showed resistance to at least one drug and 77.1% were multiresistant. The most effective drug was oxacillin. The study reports the alarming antimicrobial resistance of Staphylococcus isolated from canine pyoderma samples.O estudo se propõe a avaliar a etiologia e a susceptibilidade de estafilococos isolados de cães com dermatite. Os isolados foram identificados por provas bioquímicas e testados quanto a sua susceptibilidade a 15 antimicrobianos. Trinta e nove isolados de Staphylococci foram obtidos, e S. pseudintermedius foi mais freqüente (47,4%. Todos isolados apresentaram resistência a pelo menos uma droga e 77,1% foram multirresistentes. A oxacilina foi a droga mais eficaz. O estudo demonstra níveis alarmantes de resistência antimicrobiana nos Staphylococcus de pioderma canino.

  3. Prevalence of methicillin-resistant staphylococci isolated from different biological samples at Policlinico Umberto I of Rome: correlation with vancomycin susceptibility

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    Maria Teresa Mascellino

    2011-03-01

    Full Text Available The methicillin-resistance is increasing all over the world in the last decade. It is more frequent among coagulase-negative staphylococci (MRCoNS; infact the 52% of S. epidermidis strains results to be resistant to methicillin.The methicillin-resistant strains also show a reduced sensitivity towards the first-line agents such as glycopeptides and other antibiotics commonly used in therapy such as trimethoprim-sulphamethoxazole, imipenem, gentamycin, fosfomycin and chlarytromicin. Unlike MRSA (Methicillin-resistant S. aureus, MRCoNS resistance to glycopeptides generally concerns teicoplanin. Although vancomycin resistance is rare in Staphylococcus isolates, the detected shift towards higher values of MICs might affect patient’s clinical outcome.

  4. A comparative evaluation of methicillin-resistant staphylococci isolated from harness racing-horses, breeding mares and riding-horses in Italy

    Institute of Scientific and Technical Information of China (English)

    Karina Mallardo; Sandra Nizza; Filomena Fiorito; Ugo Pagnini; Luisa De Martino

    2013-01-01

    Objective:To investigate the prevalence of methicillin-resistant staphylococci (MRS) which is a potencial risk factor of transmission between animals and humans in different types of horses (harness racing-horses, breeding mares and riding-horses) and to compare the antimicrobial resistance of the isolates. Methods:A total of 191 healthy horses, housed at different locations of the Campania Region (Italy), were included in the study. Nasal swab samples were collected from each nostril of the horses. The mecA gene was detected by a nested PCR technique. Antibiotic susceptibility was tested for each isolate. Results: MRS was isolated from nasal samples of 68/191 (35.6%; 95% CI: 28.9%-42.9%) healthy horses. All isolates were coagulase-negative with the exception of two coagulase-positive MRS strains, identified as Staphylococcus aureus and Staphylococcus pseudintermedius, 2/83 (2.4%; 95%CI: 0.4%-9.2%). Interestingly, both coagulase-positive MRS isolates were from harness racing-horses. These horses also presented a significantly higher positivity for MRS (53.3%; 95%CI: 40.1%-66.1%) than the breeding mares and riding-horses groups. Antibiotic susceptibility testing showed difference between isolates due to different origins except for an almost common high resistance to aminopenicillins, such as ampicillin and amoxicillin. Conclusions:It can be concluded that harness racing-horses may act as a significant reservoir of MRS as compared to breeding mares and riding-horses.

  5. A comparative evaluation of methicillin-resistant staphylococci isolated from harness racing-horses, breeding mares and riding-horses in Italy.

    Science.gov (United States)

    Mallardo, Karina; Nizza, Sandra; Fiorito, Filomena; Pagnini, Ugo; De Martino, Luisa

    2013-03-01

    To investigate the prevalence of methicillin-resistant staphylococci (MRS) which is a potencial risk factor of transmission between animals and humans in different types of horses (harness racing-horses, breeding mares and riding-horses) and to compare the antimicrobial resistance of the isolates. A total of 191 healthy horses, housed at different locations of the Campania Region (Italy), were included in the study. Nasal swab samples were collected from each nostril of the horses. The mecA gene was detected by a nested PCR technique. Antibiotic susceptibility was tested for each isolate. MRS was isolated from nasal samples of 68/191 (35.6%; 95% CI: 28.9%-42.9%) healthy horses. All isolates were coagulase-negative with the exception of two coagulase-positive MRS strains, identified as Staphylococcus aureus and Staphylococcus pseudintermedius, 2/83 (2.4%; 95% CI: 0.4%-9.2%). Interestingly, both coagulase-positive MRS isolates were from harness racing-horses. These horses also presented a significantly higher positivity for MRS (53.3%; 95% CI: 40.1%-66.1%) than the breeding mares and riding-horses groups. Antibiotic susceptibility testing showed difference between isolates due to different origins except for an almost common high resistance to aminopenicillins, such as ampicillin and amoxicillin. It can be concluded that harness racing-horses may act as a significant reservoir of MRS as compared to breeding mares and riding-horses.

  6. Epidemiology of coagulase-negative staphylococci intramammary infection in dairy cattle and the effect of bacteriological culture misclassification.

    Science.gov (United States)

    Dufour, S; Dohoo, I R; Barkema, H W; Descôteaux, L; Devries, T J; Reyher, K K; Roy, J-P; Scholl, D T

    2012-06-01

    Objectives of this study were to identify the manageable risk factors associated with the lactational incidence, elimination, and prevalence of coagulase-negative staphylococci (CNS) intramammary infections (IMI) while taking into account the difficulties inherent to their diagnosis. A second objective was to evaluate the effect of CNS IMI misclassification in mastitis research. A cohort of 90 Canadian dairy herds was followed throughout 2007 to 2008. In each herd, series of quarter milk samples were collected from a subsample of cows and bacteriological culture was performed to identify prevalent, incident, and eliminated CNS IMI. Practices used on farms were captured using direct observations and a validated questionnaire. The relationships between herd CNS IMI prevalence and herd incidence and elimination rates were explored using linear regression. Manageable risk factors associated with the prevalence, incidence, or elimination of CNS IMI were identified via Bayesian analyses using a latent class model approach, allowing adjustment of the estimates for the imperfect sensitivity and specificity of bacteriological culture. After adjustment for the diagnostic test limitations, a mean CNS IMI quarter prevalence of 42.7% [95% confidence interval (CI): 34.7, 50.1] and incidence and elimination rates of 0.29 new IMI/quarter-month (95% CI: 0.21, 0.37) and 0.79 eliminated IMI/quarter-month (95% CI: 0.66, 0.91), respectively, were observed. Considerable biases of the estimates were observed when CNS IMI misclassification was ignored. These biases were important for measures of association with risk factors, were almost always toward the null value, and led to both type I and type II errors. Coagulase-negative staphylococci IMI incidence appeared to be a stronger determinant of herd IMI prevalence than IMI elimination rate. The majority of herds followed were already using blanket dry cow treatment and postmilking teat disinfection. A holistic approach considering

  7. Binding to extracellular matrix proteins and formation of biogenic amines by food-associated coagulase-negative staphylococci.

    Science.gov (United States)

    Seitter, Marion; Geng, Bettina; Hertel, Christian

    2011-02-28

    In connection with a study on the DNA microarray based detection of genes involved in safety and technologically relevant properties (Seitter (née Resch) et al., 2011), food-associated coagulase-negative staphylococci (CNS) were investigated phenotypically with regard to their ability to bind to the extracellular matrix proteins (ECM) and to produce biogenic amines. The properties have been shown to be involved in the colonization of injured tissue and invasion into host cells as well as in pharmacologic effects on humans, respectively. The CNS exhibited a low, but nevertheless clearly measurable ECM binding capacity, except for strains of Staphylococcus equorum and Staphylococcus succinus, which show a comparable or even higher binding to fibrinogen and fibronectin than that of the control strain Staphylococcus aureus Cowan. Formation of biogenic amines could be often detected in S. carnosus, S. condimenti and S. strains, but rarely in S. equorum and not in S. succinus and S. xylosus strains. Mostly, 2-phenylethylamine, tyramine and tryptamine were formed by resting cells in amounts 100 mg/l) of 2-phenylethylamine and putrescine. This study confirmed the need of consideration of ECM binding and biogenic amine formation in the safety assessment of CNS used in the production of fermented foods.

  8. Evidence for inter- and intraspecies biofilm formation variability among a small group of coagulase-negative staphylococci.

    Science.gov (United States)

    Oliveira, Fernando; Lima, Cláudia Afonso; Brás, Susana; França, Ângela; Cerca, Nuno

    2015-10-01

    Coagulase-negative staphylococci (CoNS) are common bacterial colonizers of the human skin. They are often involved in nosocomial infections due to biofilm formation in indwelling medical devices. While biofilm formation has been extensively studied in Staphylococcus epidermidis, little is known regarding other CoNS species. Here, biofilms from six different CoNS species were characterized in terms of biofilm composition and architecture. Interestingly, the ability to form a thick biofilm was not associated with any particular species, and high variability on biofilm accumulation was found within the same species. Cell viability assays also revealed different proportions of live and dead cells within biofilms formed by different species, although this parameter was particularly similar at the intraspecies level. On the other hand, biofilm disruption assays demonstrated important inter- and intraspecies differences regarding extracellular matrix composition. Lastly, confocal laser scanning microscopy experiments confirmed this variability, highlighting important differences and common features of CoNS biofilms. We hypothesized that the biofilm formation heterogeneity observed was rather associated with biofilm matrix composition than with cells themselves. Additionally, our results indicate that polysaccharides, DNA and proteins are fundamental pieces in the process of CoNS biofilm formation.

  9. Evaluation of latex agglutination test and oxacillin resistant screening agar base (ORSAB medium for the detection of oxacillin resistant coagulase negative Staphylococci (ORCoNS (Preliminary study

    Directory of Open Access Journals (Sweden)

    Sjoekoer M. Dzen

    2007-12-01

    Full Text Available Coagulase negative Staphylococci (CoNS are recognized as an important cause of nosocomial infection, especially in neonates and patients with indwelling prosthetic devices. The CoNS resistance rate to oxacillin has been increasing. Therefore, rapid and accurate detection of oxacillin resistance is essential in order to determine the most appropriate antimicrobial therapy. This study aimed to prove that latex agglutination test and oxacillin resistant screening agar base (ORSAB medium can be used for rapid detection of oxacilllin resistant CoNS (ORCoNS. Latex agglutination test and ORSAB medium compared with the conventional method was conducted in this study toward 30 clinical isolates of CoNS for the detection of ORCoNS. Mc Nemar test was used to analyze the data. The study result revealed that there was no significant difference (P>0.05 in terms of ORCoNS detection between the latex agglutination test and ORSAB medium on the one hand, and the conventional method on the other. It is concluded that latex agglutination test and ORSAB medium can be used for rapid detection of ORCoNS. (Med J Indones 2007; 16:228-32Keywords: nosocomial infection, rapid detection, mecA gene

  10. Drug resistance and genetic characteristics of clinical isolates of staphylococci in Myanmar: high prevalence of PVL among methicillin-susceptible Staphylococcus aureus belonging to various sequence types

    Directory of Open Access Journals (Sweden)

    M.S. Aung

    2016-03-01

    Full Text Available Prevalence, drug resistance and genetic characteristics were analysed for a total of 128 clinical isolates of staphylococci obtained from a tertiary hospital in Myanmar. The dominant species were S. aureus (39% and S. haemolyticus (35%, followed by S. epidermidis (6% and S. saprophyticus (5%. The majority of S. haemolyticus isolates (71.1% harboured mecA, showing high resistance rates to ampicillin, cephalosporins, erythromycin and levofloxacin, while methicillin-resistant S. aureus (MRSA was only 8% (four isolates among S. aureus with type IV SCCmec. Panton-Valentine leukocidin (PVL genes were detected in 20 isolates of S. aureus (40%, among which only one isolate was MRSA belonging to sequence type (ST 88/agr-III/coa-IIIa, and the other 19 methicillin-susceptible S. aureus (MSSA isolates were classified into six STs (ST88, ST121, ST1153, ST1155, ST1930, ST3206. An ST1153 MSSA isolate with PVL was revealed to belong to a novel coa type, XIIIa. ST121 S. aureus was the most common in the PVL-positive MSSA (47%, 9/19, harbouring genes of bone sialoprotein and variant of elastin binding protein as a distinctive feature. Although PVL-positive MSSA was susceptible to most of the antimicrobial agents examined, ST1930 isolates were resistant to erythromycin and levofloxacin. ST59 PVL-negative MRSA and MSSA had more resistance genes than other MRSA and PVL-positive MSSA, showing resistance to more antimicrobial agents. This study indicated higher prevalence of mecA associated with multiple drug resistance in S. haemolyticus than in S. aureus, and dissemination of PVL genes to multiple clones of MSSA, with ST121 being dominant, among hospital isolates in Myanmar.

  11. Evaluation of BD MAX Staph SR Assay for Differentiating Between Staphylococcus aureus and Coagulase-Negative Staphylococci and Determining Methicillin Resistance Directly From Positive Blood Cultures.

    Science.gov (United States)

    Lee, Jaewoong; Park, Yeon Joon; Park, Dong Jin; Park, Kang Gyun; Lee, Hae Kyung

    2017-01-01

    We evaluated the performance of the BD MAX StaphSR Assay (SR assay; BD, USA) for direct detection of Staphylococcus aureus and methicillin resistance not only in S. aureus but also in coagulase-negative Staphylococci (CNS) from positive blood cultures. From 228 blood culture bottles, 103 S. aureus [45 methicillin-resistant S. aureus (MRSA), 55 methicillin-susceptible S. aureus (MSSA), 3 mixed infections (1 MRSA+Enterococcus faecalis, 1 MSSA+MRCNS, 1 MSSA+MSCNS)], and 125 CNS (102 MRCNS, 23 MSCNS) were identified by Vitek 2. For further analysis, we obtained the cycle threshold (Ct) values from the BD MAX system software to determine an appropriate cutoff value. For discrepancy analysis, conventional mecA/mecC PCR and oxacillin minimum inhibitory concentrations (MICs) were determined. Compared to Vitek 2, the SR assay identified all 103 S. aureus isolates correctly but failed to detect methicillin resistance in three MRSA isolates. All 55 MSSA isolates were correctly identified by the SR assay. In the concordant cases, the highest Ct values for nuc, mecA, and mec right-extremity junction (MREJ) were 25.6, 22, and 22.2, respectively. Therefore, we selected Ct values from 0-27 as a range of positivity, and applying this cutoff, the sensitivity/specificity of the SR assay were 100%/100% for detecting S. aureus, and 97.9%/98.1% and 99.0%/95.8% for detecting methicillin resistance in S. aureus and CNS, respectively. We propose a Ct cutoff value for nuc/mec assay without considering MREJ because mixed cultures of MSSA and MRCNS were very rare (0.4%) in the positive blood cultures.

  12. Diversity and evolution of blaZ from Staphylococcus aureus and coagulase-negative staphylococci

    DEFF Research Database (Denmark)

    Olsen, John E.; Christensen, Henrik; Aarestrup, Frank Møller

    2006-01-01

    NS) and Staphylococcus aureus of bovine origin. Methods: blaZ was detected in 143 strains of penicillin-resistant S. aureus and CoNS from five Danish cattle herds (n = 25/23), random CoNS isolates from Denmark (n = 37), a collection of S. aureus from six different countries (n = 52), humans in Denmark (n = 3) and beta...... and sequences retrieved from public databases were compared. A phylogenetic tree showed that blaZ exists in three evolutionary lines: one group was of plasmid origin, one group was of chromosomal origin and one intermediate group. Sixty-nine sequence types were demonstrated. They translated into 11 BlaZ protein...... types. The major types all contained strains of both human and bovine origin, and more than one Staphylococcus species, demonstrating a shared gene pool. In a comparison of S. aureus and CoNS obtained from five Danish cattle herds, the same type of blaZ was only detected in one case. Conclusions...

  13. Effect of parenteral nutrition solutions on biofilm formation of coagulase-negative Staphylococci: An experimental study

    Directory of Open Access Journals (Sweden)

    J. Sedef Göçmen

    2012-12-01

    Full Text Available Objectives: In our study we investigated the effects ofparenteral nutrition (PN solutions on Coagulase negativestaphylococci (CoNS biofilm production.Materials and methods: Thirty nine CoNS strains isolatedfrom hemocultures and a reference strain (ATCC 12228Staphylococcus epidermidis were included. Bacterial dilutionswere made in Tryptic Soy Broth (TSB. The experimentalmediums were 1. Glucose, 2. Amino acid, 3. Lipid,4. Glucose+ Amino acid+ lipid, 5. Glucose+ Amino acid, 6.Glucose+ Lipid, 7. Amino acid+ Lipid, and 8. Control (TSB.Biofilm formation was evaluated by “quantitative microdilutionplaque test”. The values greater than cut off valueare considered as positive. Biofilm positivity was dividedinto 3 groups (mild, moderate and intensive and all otherstrains under cutoff value were accepted as negative. Thenumbers of biofilm positive strains derived from 1-7. mediumswere compared with each other, and with the resultsof control.Results: The three-component PN solution and two componentPN solutions containing glucose+ lipid and aminoacid+ lipid were found to increase the biofilm productionactivity of CoNS when compared to the control group.Slime positivity in medium 1 and 2 was lower than controlsignificantly, in medium 4, 6, and 7 slime positivity washigher considerably. The indifferent results were obtainedwithin the mediums 1, 2, 3 and within the mediums 4, 5, 6,and 7.Conclusions: In our study, it was found that, glucose, aminoacid and lipid solutions which were building structuresof PN decreased the biofilm production when used solitary.However use of the compounds increased the biofilmproduction. Therefore, we can conclude that PN solutionsgiven as mixtures in routine practice increase the risk ofcatheter infection. J Clin Exp Invest 2012; 3(4: 505-509Key words: Catheter-related infections, biofilm, parenteralnutrition

  14. Exploring the metabolic heterogeneity of coagulase-negative staphylococci to improve the quality and safety of fermented meats: a review.

    Science.gov (United States)

    Sánchez Mainar, María; Stavropoulou, Despoina Angeliki; Leroy, Frédéric

    2017-04-17

    The production of fermented meats, such as fermented sausage, relies on the metabolic activities of lactic acid bacteria and catalase-positive cocci, in particular the group of coagulase-negative staphylococci (CNS). Conventional use of CNS as meat starter cultures usually leads to an appropriate cured colour development based on their nitrate reductase activity, whereas their catalase activity reduces oxidative damage. In addition, CNS metabolism contributes to flavour, although the precise effects are difficult to estimate. There are reasons to believe that these basic technological features of CNS can be further enlarged by exploring their full metabolic potential. Non-negligible differences in metabolism among and within different species of CNS indicate that a rational selection of strains may lead to the development of novel starter cultures with enhanced functionality. Firstly, the use of CNS strains with a superior ability to use alternative energy sources, such as nucleosides or arginine, may improve culture competitiveness and survival. Secondly, cured colour generation could be optimised to lower the amounts of curing salts needed, either by selecting for efficient nitrate-reducing CNS strains or by exploring the potential alternative based on nitric oxide synthase activity. Thirdly, CNS with specific aroma-producing abilities may help to accentuate specific flavours, whereby the selection of wild-type strains from artisan-type fermented sausages seems attractive in the framework of innovation-through-tradition. Finally, bacteriocin-producing CNS strains may offer solutions for bioprotection towards meat pathogens such as Clostridium botulinum and Staphylococcus aureus. Overall, making use of the metabolic inter- and intraspecies heterogeneity of CNS is promising for the elaboration of healthier, tastier, and safer fermented meats. Yet, the proposed strategies are sometimes still overly theoretical and speculative, requiring further proof-of-principle.

  15. Identification of ABC transporter genes conferring combined pleuromutilin-lincosamide-streptogramin A resistance in bovine methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci.

    Science.gov (United States)

    Wendlandt, Sarah; Kadlec, Kristina; Feßler, Andrea T; Schwarz, Stefan

    2015-06-12

    The aim of this study was to investigate the genetic basis of combined pleuromutilin-lincosamide-streptogramin A resistance in 26 unrelated methicillin-resistant Staphylococcus aureus (MRSA) and coagulase-negative staphylococci (CoNS) from dairy cows suffering from mastitis. The 26 pleuromutilin-resistant staphylococcal isolates were screened for the presence of the genes vga(A), vga(B), vga(C), vga(E), vga(E) variant, sal(A), vmlR, cfr, lsa(A), lsa(B), lsa(C), and lsa(E) by PCR. None of the 26 isolates carried the genes vga(B), vga(C), vga(E), vga(E) variant, vmlR, cfr, lsa(A), lsa(B), or lsa(C). Two Staphylococcus haemolyticus and single Staphylococcus xylosus, Staphylococcus lentus, and Staphylococcus hominis were vga(A)-positive. Twelve S. aureus, two Staphylococcus warneri, as well as single S. lentus and S. xylosus carried the lsa(E) gene. Moreover, single S. aureus, S. haemolyticus, S. xylosus, and Staphylococcus epidermidis were positive for both genes, vga(A) and lsa(E). The sal(A) gene was found in a single Staphylococcus sciuri. All ABC transporter genes were located in the chromosomal DNA, except for a plasmid-borne vga(A) gene in the S. epidermidis isolate. The genetic environment of the lsa(E)-positive isolates was analyzed using previously described PCR assays. Except for the S. warneri and S. xylosus, all lsa(E)-positive isolates harbored a part of the previously described enterococcal multiresistance gene cluster. This is the first report of the novel lsa(E) gene in the aforementioned bovine CoNS species. This is also the first identification of the sal(A) gene in a S. sciuri from a case of bovine mastitis. Moreover, the sal(A) gene was shown to also confer pleuromutilin resistance. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Technical note: a pilot study using a mouse mastitis model to study differences between bovine associated coagulase-negative staphylococci.

    Science.gov (United States)

    Breyne, K; De Vliegher, S; De Visscher, A; Piepers, S; Meyer, E

    2015-02-01

    Coagulase-negative staphylococci (CNS) are a group of bacteria classified as either minor mastitis pathogens or commensal microbiota. Recent research suggests species- and even strain-related epidemiological and genetic differences within the large CNS group. The current pilot study investigated in 2 experiments whether a mouse mastitis model validated for bovine Staphylococcus aureus can be used to explore further differences between CNS species and strains. In a first dose titration experiment, a low inoculum dose of S. aureus Newbould 305 (positive control) was compared with increasing inoculum doses of a Staphylococcus chromogenes strain originating from a chronic bovine intramammary infection to a sham-inoculated mammary glands (negative control). In contrast to the high bacterial growth following inoculation with S. aureus, S. chromogenes was retrieved in very low levels at 24 h postinduction (p.i.). In a second experiment, the inflammation inflicted by 3 CNS strains was studied in mice. The host immune response induced by the S. chromogenes intramammary strain was compared with the one induced by a Staphylococcus fleurettii strain originating from cow bedding sawdust and by a S. chromogenes strain originating from a teat apex of a heifer. As expected, at 28 and 48 h p.i., low bacterial growth and local neutrophil influx in the mammary gland were induced by all CNS strains. As hypothesized, bacterial growth p.i. was the lowest for S. fleurettii compared with that induced by the 2 S. chromogenes strains, and the overall immune response established by the 3 CNS strains was less pronounced compared with the one induced by S. aureus. Proinflammatory cytokine profiling revealed that S. aureus locally induced IL-6 and IL-1β but not TNF-α, whereas, overall, CNS-inoculated glands lacked a strong cytokine host response but also induced IL-1β locally. Compared with both other CNS strains, S. chromogenes from the teat apex inflicted a more variable IL-1β response

  17. Concentrations of procalcitonin and C-reactive protein, white blood cell count, and the immature-to-total neutrophil ratio in the blood of neonates with nosocomial infections: Gram-negative bacilli vs coagulase-negative staphylococci

    OpenAIRE

    Kordekag, A.

    2011-01-01

    Abstract This study was undertaken to determine whether concentrations of procalcitonin in the blood of neonates with nosocomial infections depend on the type of pathogen. Qualification for the study group was based on the clinical signs of infection. We found that infections with Gram-positive (chiefly coagulase-negative staphylococci) and Gram-negative bacteria are accompanied by elevated concentrations of procalcitonin. In the case of Gram-positive bacteria, other laboratory sig...

  18. Poor value of surveillance cultures for prediction of septicaemia caused by coagulase-negative staphylococci in patients undergoing haemodialysis with central venous catheters

    DEFF Research Database (Denmark)

    Nielsen, J; Kolmos, H J; Rosdahl, V T

    1998-01-01

    Surveillance cultures for the demonstration of coagulase-negative staphylococci in patients on catheter haemodialysis were performed in an attempt to predict dialysis catheter-related septicaemia. In all, 43 patients with 67 haemodialysis catheters were followed for a 1-y period. Once a week, swab...... was cultured from the skin, hub and catheter blood, respectively. In two-thirds of all catheter periods, S. epidermidis was cultured from catheter blood despite lack of clinical signs of septicaemia. In conclusion, cultures from the skin, hub or catheter blood are not useful for prediction of dialysis catheter-related...

  19. Coagulase-negative staphylococci: update on the molecular epidemiology and clinical presentation, with a focus on Staphylococcus epidermidis and Staphylococcus saprophyticus.

    Science.gov (United States)

    Widerström, M; Wiström, J; Sjöstedt, A; Monsen, T

    2012-01-01

    Coagulase-negative staphylococci (CoNS), originally described as ubiquitous commensals of the healthy human skin and mucosa, have emerged as important opportunistic pathogens primarily causing healthcare-associated infections in patients with indwelling medical devices. Recent studies, utilizing new molecular typing methods, particularly on Staphylococcus epidermidis, have increased our understanding of the mechanisms that contribute to the evolutionary success of these extremely versatile microorganisms. In the following mini-review, we summarize recent research in this area focusing on the molecular methods and epidemiology of S. epidermidis and S. saprophyticus.

  20. First identification of methicillin-resistant Staphylococcus pseudintermedius strains among coagulase-positive staphylococci isolated from dogs with otitis externa in Trinidad, West Indies

    Directory of Open Access Journals (Sweden)

    Francis Dziva

    2015-12-01

    Full Text Available Background: Otitis externa is a common inflammatory ear disease in dogs caused by a variety of pathogens, and coagulase-positive staphylococci are frequently isolated from such infections. Objective: To identify antimicrobial susceptibility profiles and methicillin-resistant strains among coagulase-positive staphylococci isolated from otitis externa in dogs. Methods: A cross-sectional study was performed over 2 years on 114 client-owned dogs presented to the Veterinary Teaching Hospital with a primary complaint of ear infections. Swabs were obtained from both ears and cultured for staphylococci which were subsequently confirmed as coagulase-positive using rabbit plasma. Antimicrobial susceptibility assays were assessed on all isolates followed by subsequent genetic analysis for species identification and detection of the mecA gene. Results: Sixty-five coagulase-positive staphylococci were isolated from 114 client-owned dogs. The isolates exhibited resistance against neomycin (58.5%, streptomycin (49.2%, penicillin (49.2%, polymyxin B (44.6%, tetracycline (36.9%, sulphamethoxazole/trimethoprim (33.8%, kanamycin (33.8%, doxycycline (32.3%, norfloxacin (23.1%, amoxicillin/clavulanate (20%, ciprofloxacin (20%, enrofloxacin (18.5%, gentamicin (16.9%, and cephalothin (9.2%. Forty (61.5% of the isolates were resistant to at least three or more antimicrobials and 10 were sensitive to all. Using a multiplex polymerase chain reaction assay based on species-specific regions of the thermonuclease (nuc gene, 38/65 (58.5% isolates were classified as Staphylococcus aureus, 23/65 (35.4% as S. pseudintermedius, 2/65 (3.1% as S. intermedius, and 2/65 (3.1% as S. schleiferi. Analysis for the mecA gene revealed two positive isolates of S. pseudintermedius which were oxacillin-resistant, representing a first report of such organisms in the Caribbean. Conclusion: Despite the relatively high prevalence of multidrug-resistant coagulase-positive staphylococci in

  1. First identification of methicillin-resistant Staphylococcus pseudintermedius strains among coagulase-positive staphylococci isolated from dogs with otitis externa in Trinidad, West Indies

    Science.gov (United States)

    Dziva, Francis; Wint, Crystal; Auguste, Tennille; Heeraman, Carolyn; Dacon, Cherrelle; Yu, Priscilla; Koma, Lee M.

    2015-01-01

    Background Otitis externa is a common inflammatory ear disease in dogs caused by a variety of pathogens, and coagulase-positive staphylococci are frequently isolated from such infections. Objective To identify antimicrobial susceptibility profiles and methicillin-resistant strains among coagulase-positive staphylococci isolated from otitis externa in dogs. Methods A cross-sectional study was performed over 2 years on 114 client-owned dogs presented to the Veterinary Teaching Hospital with a primary complaint of ear infections. Swabs were obtained from both ears and cultured for staphylococci which were subsequently confirmed as coagulase-positive using rabbit plasma. Antimicrobial susceptibility assays were assessed on all isolates followed by subsequent genetic analysis for species identification and detection of the mecA gene. Results Sixty-five coagulase-positive staphylococci were isolated from 114 client-owned dogs. The isolates exhibited resistance against neomycin (58.5%), streptomycin (49.2%), penicillin (49.2%), polymyxin B (44.6%), tetracycline (36.9%), sulphamethoxazole/trimethoprim (33.8%), kanamycin (33.8%), doxycycline (32.3%), norfloxacin (23.1%), amoxicillin/clavulanate (20%), ciprofloxacin (20%), enrofloxacin (18.5%), gentamicin (16.9%), and cephalothin (9.2%). Forty (61.5%) of the isolates were resistant to at least three or more antimicrobials and 10 were sensitive to all. Using a multiplex polymerase chain reaction assay based on species-specific regions of the thermonuclease (nuc) gene, 38/65 (58.5%) isolates were classified as Staphylococcus aureus, 23/65 (35.4%) as S. pseudintermedius, 2/65 (3.1%) as S. intermedius, and 2/65 (3.1%) as S. schleiferi. Analysis for the mecA gene revealed two positive isolates of S. pseudintermedius which were oxacillin-resistant, representing a first report of such organisms in the Caribbean. Conclusion Despite the relatively high prevalence of multidrug-resistant coagulase-positive staphylococci in Trinidad

  2. Utilization of horse plasma in coagulase test for Staphylococci isolated from raw milk/ Utilização de plasma de cavalo no teste de coagulase em Estafilococos isolados de leite cru

    Directory of Open Access Journals (Sweden)

    Luciane Bilia de Moraes

    2005-06-01

    Full Text Available Staphylococci are frequently related to food poisoning outbreaks that are caused by their enterotoxins. The enterotoxins production is highly correlated to coagulase production, which one is detected in identification of pathogenic strains. Usually the coagulase test is conducted using lyophilized rabbit plasma, or even fresh rabbit blood, separated by sedimentation. The lyophilized plasma has high costs and the fresh material is obtained in small volumes from the animals, what made it unpractical. The objective of this work was evaluating the utilization of an alternative material to rabbit plasma, in this case the horse plasma, in coagulase test for staphylococci isolated from raw milk. So, 410 typical and atypical staphylococci colonies were tested for coagulase production using both plasmas. The results showed 284 (69.26% positives for coagulase production and 126 (30.73% negatives, with 100% of concordance between both plasmas. These results indicate the viability in using horse plasma in coagulase test for staphylococci isolated from raw milk, and complementary studies are necessary with strains from different types of food.Estafilococos têm grande importância na indústria de alimentos por serem associados a surtos de intoxicação alimentar, pela ação de suas enterotoxinas. A produção dessas substâncias tem alta correlação com a produção da enzima coagulase, que é detectada na identificação de cepas potencialmente patogênicas. Esse teste é tradicionalmente realizado com plasma de coelho liofilizado ou pela sua separação de sangue coletado de coelhos. O plasma liofilizado tem alto custo e a punção cardíaca, além de ser bastante cruenta, permite a obtenção apenas de pequenos volumes de material. Este trabalho tem como objetivo verificar a viabilidade da utilização de um material alternativo ao plasma de coelho, neste caso o plasma de cavalo, no teste de coagulase para estafilococos isolados de leite cru. Assim

  3. Isolation of coagulase-positive staphylococci from bitches' colostrum and milk and genetic typing of methicillin-resistant Staphylococcus pseudintermedius strains.

    Science.gov (United States)

    Rota, Ada; Corrò, Michela; Drigo, Ilenia; Bortolami, Alessio; Börjesson, Stefan

    2015-07-23

    Among the coagulase-positive, potentially pathogenic staphylococci, Staphylococcus pseudintermedius has been frequently isolated from bitches' milk. This organism colonizes the mammary gland or causes infection, while S. aureus has been only occasionally reported. The objective of this study was to investigate the occurrence and persistence of coagulase-positive staphylococci in the colostrum and milk of postpartum bitches, either treated or untreated with antimicrobials, and to assess the incidence, antibiotic resistance profile and genetic type of the methicillin-resistant strains. On postpartum D1, D7 and D15, drops of secretion were collected from the mammary glands of 27 postpartum bitches, nine of which were treated with antimicrobials. Coagulase-positive staphylococci were identified, antimicrobial susceptibility and the presence of mecA were tested and the genetic profile of methicillin-resistant strains was assessed. Staphylococcus pseudintermedius was the only coagulase-positive staphylococcus isolated, and its presence was detected in 21 out of 27 bitches and in 66 out of 145 swabs. In a single bitch, it caused puerperal mastitis. In untreated bitches, the frequency of isolation was lower in colostrum than in milk. All of the isolates except one were resistant to at least three antimicrobial classes, while 14 out of 66 S. pseudintermedius strains were methicillin-resistant mecA positive (MRSP) and were isolated from eight bitches housed in the same breeding kennel. A significant association was found between antimicrobial treatment and the presence of MRSP. Six of the 12 typed isolates belonged to spa-type t02 carrying SCCmec II/III, and another six were non-typeable with spa carrying SCCmec IV. The t02-SCCmec II/III isolates were sequence type (ST) 71; four NT-SCCmec IV isolates were ST258 and two were ST369. PFGE showed that isolates from the same dog had identical band patterns, while isolates from different dogs had unique band patterns. MRSP strains

  4. Rapid differentiation of Staphylococcus aureus, Staphylococcus epidermidis and other coagulase-negative staphylococci and meticillin susceptibility testing directly from growth-positive blood cultures by multiplex real-time PCR.

    Science.gov (United States)

    Jukes, Leanne; Mikhail, Jane; Bome-Mannathoko, Naledi; Hadfield, Stephen J; Harris, Llinos G; El-Bouri, Khalid; Davies, Angharad P; Mack, Dietrich

    2010-12-01

    This study evaluated a multiplex real-time PCR method specific for the mecA, femA-SA and femA-SE genes for rapid identification of Staphylococcus aureus, Staphylococcus epidermidis and non-S. epidermidis coagulase-negative staphylococci (CoNS), and meticillin susceptibility testing directly in positive blood cultures that grew Gram-positive cocci in clusters. A total of 100 positive blood cultures produced: 39 S. aureus [12 meticillin-resistant S. aureus (MRSA), 31% of all the S. aureus]; 30 S. epidermidis (56.6% of the CoNS), 8 Staphylococcus capitis (15.1%), 3 Staphylococcus saprophyticus (5.7%), 4 Staphylococcus hominis (7.5%), 3 Staphylococcus haemolyticus (5.7%), 2 Staphylococcus warneri (3.8%), 1 Staphylococcus cohnii (1.9%) and 2 unidentified Staphylococcus spp. (3.8%); and 1 Micrococcus luteus in pure culture. Two blood cultures had no growth on subculture and five blood cultures grew mixed CoNS. For the 95 blood cultures with pure growth or no growth on subculture, there was very good agreement between real-time PCR and the BD Phoenix identification system for staphylococcal species categorization in S. aureus, S. epidermidis and non-S. epidermidis CoNS and meticillin-resistance determination (Cohen's unweighted kappa coefficient κ=0.882). All MRSA and meticillin-susceptible S. aureus were correctly identified by mecA amplification. PCR amplification of mecA was more sensitive for direct detection of meticillin-resistant CoNS in positive blood cultures than testing with the BD Phoenix system. There were no major errors when identifying staphylococcal isolates and their meticillin susceptibility within 2.5 h. Further studies are needed to evaluate the clinical benefit of using such a rapid test on the consumption of glycopeptide antibiotics and the alteration of empiric therapy in the situation of positive blood cultures growing staphylococci, and the respective clinical outcomes.

  5. Increasing Resistance of Coagulase-Negative Staphylococci in Total Hip Arthroplasty Infections: 278 THA-Revisions due to Infection Reported to the Norwegian Arthroplasty Register from 1993 to 2007

    Directory of Open Access Journals (Sweden)

    Olav Lutro

    2014-01-01

    Full Text Available We investigated bacterial findings from intraoperative tissue samples taken during revision due to infection after total hip arthroplasty (THA. The aim was to investigate whether the susceptibility patterns changed during the period from 1993 through 2007. Reported revisions due to infection in the Norwegian Arthroplasty Register (NAR were identified, and 10 representative hospitals in Norway were visited. All relevant information on patients reported to the NAR for a revision due to infection, including bacteriological findings, was collected from the medical records. A total of 278 revision surgeries with bacterial growth in more than 2 samples were identified and included. Differences between three 5-year time periods were tested by the chi-square test for linear trend. The most frequent isolates were coagulase-negative staphylococci (CoNS (41%, 113/278 and Staphylococcus aureus (19%, 53/278. The proportion of CoNS resistant to the methicillin-group increased from 57% (16/28 in the first period, 1993–1997, to 84% (52/62 in the last period, 2003–2007 (P = 0.003. There was also significant increase in resistance for CoNS to cotrimoxazole, quinolones, clindamycin, and macrolides. All S. aureus isolates were sensitive to both the methicillin-group and the aminoglycosides. For the other bacteria identified no changes in susceptibility patterns were found.

  6. Concentrations of procalcitonin and C-reactive protein, white blood cell count, and the immature-to-total neutrophil ratio in the blood of neonates with nosocomial infections: Gram-negative bacilli vs coagulase-negative staphylococci.

    Science.gov (United States)

    Kordek, A

    2011-03-01

    This study was undertaken to determine whether concentrations of procalcitonin in the blood of neonates with nosocomial infections depend on the type of pathogen. Qualification for the study group was based on the clinical signs of infection. We found that infections with Gram-positive (chiefly coagulase-negative staphylococci) and Gram-negative bacteria are accompanied by elevated concentrations of procalcitonin. In the case of Gram-positive bacteria, other laboratory signs of infection studied by us (concentration of C-reactive protein, white blood cell count, immature-to-total neutrophil ratio) were not discriminatory, confirming the diagnostic usefulness of procalcitonin measurements in nosocomial infections of the neonate with Gram-negative or Gram-positive bacteria.

  7. [Antagonism of lactobacilli, oral streptococci and staphylococci].

    Science.gov (United States)

    Chervinets, Iu V; Beliaeva, E A; Ganina, E B; Troshin, A V; Chervinets, A V

    2015-01-01

    From the oral cavity of healthy young people aged 18-22 years there were isolated 26 strains of lactobacilli, 28 streptococci, including the pathogenic and opportunistic strains, and 32 strains of staphylococci, 10 of which were methicillin-resistant S.aureus. Oral lactobacilli possessed by a high probiotic potential, showing high antagonism to methicillin-resistant staphylococci, pathogenic and opportunistic streptococci and enterococci. Oral lactobacilli showed medium and high adhesive activity that determines their high adaptive capacity. Staphylococci and streptococci in 90.3% of cases have not an antagonistic effect on lactobacilli. Isolated lactobacilli can be used as probiotic strains for oral administration.

  8. Preoperative screening cultures in the identification of staphylococci causing wound and valvular infections in cardiac surgery

    NARCIS (Netherlands)

    E. J. Ridgway; A.P.R. Wilson; M. C. Kelsey

    1990-01-01

    textabstractCultures of nasal or presternal swabs form part of the routine preoperative screening of patients on the cardiac surgical ward. During a trial of antibiotic prophylaxis in 314 patients, preoperative isolates of Staphylococcus aureus and coagulase-negative staphylococci were compared with

  9. 凝固酶阴性葡萄球菌菌血症临床感染特征及耐药性分析%Coagulase-negative Staphylococci bacteremia clinical infection characteristics and drug resistance analysis

    Institute of Scientific and Technical Information of China (English)

    陈倩; 孙亚娟; 郭燕菊; 王会中

    2012-01-01

    Objective To investigate the clinical characteristics of patients with coagulase-negative staphylococci (CNS) bactere-mia and the drug resistance of isolates to provide basis for clinical diagnosis and treatment. Methods Medical records between Nov. 2009 and Oct. 2011 of 56 cases with CNS-induced bacteremia in this hospital were retrospectively analyzed. Results All of the cases were with fever and leukocytosis,treatment of a variety of catheter, pre-used antibiotics and positive blood culture alarm time less than 48 hours. Of the 56 strains of CNS,82. 5% were methicillin-resistant coagulase-negative the staphylococci(MRCNS) with rel-atively high sensitivity to vancomycin, teicoplanin and rifampin. The clinical treatment efficiency was 57. 1% and the mortality rate was 10. 7%. Conclusion CNS might be an important pathogen of bloodstream infections. To identify whether CNS is pathogen might be the key for clinical diagnosis. Vancomycin could still be preferred for the treatment of MRCNS bloodstream infection.%目的 分析凝固酶阴性葡萄球菌(CNS)菌血症患者临床资料及菌株耐药性,为临床诊断与治疗提供依据.方法 对该院2009年11月至2011年10月56例CNS菌血症患者临床资料进行回顾性统计分析.结果 CNS患者均有发热和白细胞升高、植入各种留置导管、先期使用过抗菌药物、血培养阳性报警时间小于48 h;56株CNS中,82.5%为甲氧西林耐药凝固酶阴性葡萄球菌(MRCNS),对万古霉素、替考拉宁、利福平较为敏感;临床治疗有效率57.1%、死亡率10.7%.结论 CNS已是菌血症的重要病原菌,正确鉴定CNS是否为病原菌是临床诊断的关键,万古霉素仍是治疗MRCNS菌血症首选药物.

  10. Dynamic isolation technologies in negative pressure isolation wards

    CERN Document Server

    Xu, Zhonglin

    2017-01-01

    This book presents novel design principles and technologies for dynamic isolation based on experimental studies. These approaches have now become the local standard in Beijing and are currently being promoted for use nationwide. Further, the book provides details of measures and guidelines for the design process. Departing from the traditional understanding that isolation wards should be designed with high negative pressure, airtight doors and fresh air, it establishes the basis for designing biological clean rooms, including isolation wards, using a simple and convenient scientific approach. This book is intended for designers, engineers, researchers, hospital management staff and graduate students in heating ventilation air conditioning (HVAC), air cleaning technologies and related areas.

  11. 凝固酶阴性葡萄球菌菌种鉴定与苯唑西林耐药凝固酶阴性葡萄球菌检测准确性%Correlation between species-level identification of coagulase-negative staphylococci and accuracy of methicillin resistance coagulase-negative staphylococci detection

    Institute of Scientific and Technical Information of China (English)

    辜依海; 郭秀荣; 张军; 武建; 李小波; 柴阳; 王克

    2009-01-01

    Objective To evaluate accuracy of cefoxitin disk testing for detecting oxacillin resistance coagulase-negative staphylococci (MRCNS). Methods 139 clinical coagulase-negative staphylococci (CNS) were detected with ID32 STAPH. Cefoxitin disk and oxacillin disk testing were used to detect MRCNS. PBP2a was tested by latex agglutination us a reference method. Results 139 CNS isolates were identified to 8 species: Staphylococcus haemolyticus , S. epidermidis , S. hominis , S. xylosus , S. saprophyticus , S. auricularis , S. simulans and S. warneri. The sensitivity and specificity for cefoxtin disk and oxacillin disk testing were 99.0% vs. 86.0% and 91.7% vs. 74.4%, respectively. One S. epidermidis strain was identified to affect the sensitivity of cefoxitin disk testing. S. xylosus, S. warned, and S. saprophyticus were major species related to the decrease of specificity of cefoxitin disk testing. S. haemolyticus, S. hominis, S. simulans and S. auricularis were major species related to the decrease of sensitivity of oxacillin disk testing. And the decrease of specificity of oxacillin disk testing were mainly related to S. hominis , S. simulans , S. xylosus , S. auricularis , S. saprophyticus and S. warneri. Conclusions The accuracy of MRCNS detection by cefoxitin disk testing is varied due to different CNS species. So it is necessary to test PBP2a or mecA gene according to CNS species, especially for S. xylosus, S. warned and S. saprophyticus.%目的 评价凝固酶阴性葡萄球菌(CNS)菌种鉴定与苯唑西林耐药凝固酶阴件葡萄球菌(MRCNS)检测的准确性.方法 139株临床分离CNS,经ID 32 STAPH鉴定到种,用头孢西丁(FOX)、苯唑西林(OXA)纸片扩散法检测MRCNS,以Slidex MRSA detection乳胶凝集法检测青霉素结合蛋白2a(PBP2a)作为参考方法.结果 139株CNS鉴定为8个种,依次为溶血葡萄球菌、表皮葡萄球菌、人葡萄球菌、木糖葡萄球菌、腐生葡萄球菌、耳葡萄球菌、模仿葡萄球菌、沃

  12. Antimicrobial susceptibility and distribution of antimicrobial-resistance genes among Enterococcus and coagulase-negative Staphylococcus isolates recovered from poultry litter.

    Science.gov (United States)

    Simjee, Shabbir; McDermott, Patrick F; White, David G; Hofacre, Charles; Berghaus, Roy D; Carter, Peggy J; Stewart, Leigh; Liu, Tongrui; Maier, Marie; Maurer, John J

    2007-12-01

    Data on the prevalence of antimicrobial resistant enterococci and staphylococci from the poultry production environment are sparse in the United States. This information is needed for science-based risk assessments of antimicrobial use in animal husbandry and potential public-health consequences. In this study, we assessed the susceptibility of staphylococci and enterococci isolated from poultry litter, recovered from 24 farms across Georgia, to several antimicrobials of veterinary and human health importance. Among the 90 Enterococcus isolates recovered, E. hirae (46%) was the most frequently encountered species, followed by E. faecium (27%), E. gallinarum (12%), and E. faecalis (10%). Antimicrobial resistance was most often observed to tetracycline (96%), followed by clindamycin (90%), quinupristin-dalfopristin (62%), penicillin (53%), erythromycin (50%), nitrofurantoin (49%), and clarithromycin (48%). Among the 110 staphylococci isolates recovered, only coagulase-negative staphylococci (CNS) were identified with the predominant Staphylococcus species being S. sciuri (38%), S. lentus (21%), S. xylosus (14%) and S. simulans (12%). Resistance was less-frequently observed among the Staphylococcus isolates for the majority of antimicrobials tested, as compared with Enterococcus isolates, and was primarily limited to clarithromycin (71%), erythromycin (71%), clindamycin (48%), and tetracycline (38%). Multidrug resistance (MDR) phenotypes were prevalent in both Enterococcus and Staphylococcus; however, Enterococcus exhibited a statistically significant difference in the median number of antimicrobials to which resistance was observed (median = 5.0) compared with Staphylococcus species (median = 3.0). Because resistance to several of these antimicrobials in gram-positive bacteria may be attributed to the shuttling of common drug-resistance genes, we also determined which common antimicrobial-resistance genes were present in both enterococci and staphylococci. The

  13. Beta-lactamase detection in Staphylococcus aureus and coagulase-negative Staphylococcus isolated from bovine mastitis

    Directory of Open Access Journals (Sweden)

    Bruno F. Robles

    2014-04-01

    Full Text Available The objectives of the study were to evaluate the presence/production of beta-lactamases by both phenotypic and genotypic methods, verify whether results are dependent of bacteria type (Staphylococcus aureus versus coagulase-negative Staphylococcus - CNS and verify the agreement between tests. A total of 200 bacteria samples from 21 different herds were enrolled, being 100 CNS and 100 S. aureus. Beta-lactamase presence/detection was performed by different tests (PCR, clover leaf test - CLT, Nitrocefin disk, and in vitro resistance to penicillin. Results of all tests were not dependent of bacteria type (CNS or S. aureus. Several S. aureus beta-lactamase producing isolates were from the same herd. Phenotypic tests excluding in vitro resistance to penicillin showed a strong association measured by the kappa coefficient for both bacteria species. Nitrocefin and CLT are more reliable tests for detecting beta-lactamase production in staphylococci.

  14. Staphylococci with markers of antibiotic resistance collected from blood cultures

    Directory of Open Access Journals (Sweden)

    Vittorio Focarelli

    2012-06-01

    Full Text Available Introduction: Blood culture is still the gold standard for the detection of the causative agent of sepsis. Especially in intensive care patients and those with vascular catheters, the most common organisms isolated are coagulase-negative staphylococci (CoNS and Staphylococcus aureus, both characterized by multidrug resistance. Purposes of our work are the study of the incidence of markers of resistance in staphylococci and evaluation of potential changes over the years. Materials and methods: In the period January 2008-June 2011 5239 blood cultures were analyzed.They were mainly obtained from the departments of Intensive Care, Cardiology, Hematology, General Medicine, Emergency Medicine, Infectious Diseases, Oncology, Pulmonology and Pediatric Hematoncology. The vials containing the blood were incubated in the BACTEC 9120 automated tool of Becton Dickinson and susceptibility testing performed with the Phoenix instrument of the same company. Results:Within a total of 5239 blood cultures, 3967 (75.7% were negative and 1272 (24.3% positive. Fungi were isolated in 6.2% (79 of the positive ones, Gram-negative bacteria in 24.6% (313 and Gram-positive bacteria in 69.2% (880. Within the latter, 187 (21.2% were not staphylococcal isolates, 693 (78.8% were stafiloccocci mainly represented by S. epidermidis, S. aureus, S. hominis, S. haemolyticus and S. saprophyticus. Of the 693 staphylococcal isolates, 436 (62.9% were b lactamase producers, and between them 336 (77.1% were methicillin resistant, while only 3 of 436 (0.69% were S. aureus resistant to vancomycin as well.The incidence of markers of resistance was very high, especially in patients in intensive care and cardiac surgery, who are usually subjected to combined antibiotic therapy. In the three years studied there were no statistically significant differences in the resistance of staphylococci. Conclusions: The data show an alarming high number of multi-resistant staphylococci, which is often a

  15. Prevalence and molecular characterization of staphylococci isolated from sheep with subclinical mastitis in West-Azerbaijan province, Iran.

    Science.gov (United States)

    Rahman, Bentolhoda; Ownagh, Abdolghaffar; Mardani, Karim; Farrokhi Ardebili, Farhad

    2016-01-01

    This study was conducted to investigate the prevalence of subclinical mastitis caused by Staphylococcus spp. in ewes in West-Azerbaijan province of Iran. Molecular characterization of isolated Staphylococcus spp. from diseased ewes were performed using polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) and DNA sequencing of glyceraldehyde-3-phosphate dehydrogenase (gap) gene. Also, antibiotic resistance of staphylococcal isolates against different antibiotics was investigated. A total number of 900 milk samples from 450 native ewes in their mid-lactation period were examined by the California mastitis test (CMT). The CMT positive samples were cultured and bacteria were isolated from 86 (9.50%) glands and 74 (16.40%) ewes. The prevalence of subclinical mastitis in the examined ewes was 16.40%. Microbiological analysis of milk samples revealed that 27 out of 74 sheep with subclinical mastitis were infected with Staphylococcus spp. Amplification of gap gene of 27 Staphylococcus isolates generated a single amplicon of 933 bp in size confirming that isolates were belonged to Staphylococcus genus. Digestion of PCR products by AluI endonuclease generated different RFLP patterns for each species. Nucleotide sequencing of gap gene followed by phylogenetic analysis showed that the most dominant Staphylococcus species were S. epidermidis, S. xylosus and S. chromogenes. Staphylococcal isolates showed the highest resistance to penicillin and ampicillin. In conclusion, Staphylococcus species, except for the southern parts of the province, play an important role in the development of subclinical mastitis in sheep in West-Azerbaijan province of Iran. Also, chloramphenicol, ciprofloxacin and neomycin are the most effective antibiotics for treatment of this disease.

  16. Oxacillin resistance and antimicrobial susceptibility profile of Staphylococcus saprophyticus and other staphylococci isolated from patients with urinary tract infection.

    Science.gov (United States)

    Ferreira, Adriano M; Bonesso, Mariana F; Mondelli, Alessandro L; Camargo, Carlos H; Cunha, Maria de Lourdes R S

    2012-01-01

    Staphylococcus saprophyticus is the second most frequent community-acquired causative agent of urinary tract infection (UTI). The objective of this study was to evaluate the susceptibility profile and resistance detection in Staphylococcus species. isolated from patients with UTI. The isolates were investigated using the disk diffusion method, Vitek I system, E-test®, and detection of the mecA gene. Most isolates (76.2%) were resistant to oxacillin by the disk diffusion method, followed by those resistant to penicillin (72.2%). The oxacillin disk diffusion method, E-test, and Vitek I method showed higher sensitivity (94.4%) and lower specificity (28.9, 26.5, and 24.0%, respectively) than the cefoxitin disk diffusion test (sensitivity: 83.5%, specificity: 85.5%) for the detection of oxacillin resistance. The large number of oxacillin-resistant isolates indicates that the breakpoint value recommended by the Clinical and Laboratory Standards Institute may overestimate oxacillin resistance in S. saprophyticus. Thus, changes in these guidelines are necessary for the correct detection of this resistance. Copyright © 2013 S. Karger AG, Basel.

  17. Tylosin susceptibility of Staphylococci from bovine mastitis.

    Science.gov (United States)

    Entorf, Monika; Feßler, Andrea T; Kadlec, Kristina; Kaspar, Heike; Mankertz, Joachim; Peters, Thomas; Schwarz, Stefan

    2014-07-16

    Although the 16-membered macrolide tylosin is commonly used for the treatment of bovine mastitis, little information is currently available about the susceptibility of mastitis pathogens to tylosin. In the present study, 112 Staphylococcus aureus and 110 coagulase-negative Staphylococcus (CoNS) spp. isolates from cases of bovine mastitis were tested by broth microdilution and agar disk diffusion with 30 μg tylosin disks. Susceptibility to erythromycin was tested by broth microdilution and disk diffusion using 15 μg disks. Both test populations showed bimodal distributions of minimal inhibitory concentrations (MICs) and zone diameters with eleven S. aureus and eight CoNS isolates showing tylosin MICs of ≥ 256 μg/ml and no zones of growth inhibition around the tylosin 30 μg disks. All 19 isolates with tylosin MICs of ≥ 256 μg/ml were also resistant to erythromycin. For six additional erythromycin-resistant isolates, tylosin MICs of 1-8 μg/ml were observed. One S. aureus and two CoNS isolates showed inducible macrolide resistance. PCR analysis of the 25 erythromycin-resistant staphylococcal isolates identified the resistance genes erm(A), erm(B), erm(C), erm(T), mph(C) and msr(A) alone or in different combinations. An excellent correlation between the results of the different tylosin susceptibility tests (broth microdilution versus disk diffusion) was seen for S. aureus and CoNS isolates. Since tylosin does not induce the expression of the aforementioned erm genes, isolates with an inducible resistance phenotype may - if only tylosin is tested - be falsely classified as tylosin-susceptible. Thus, erythromycin should be tested in parallel and tylosin should only be used for the treatment of infections caused by erythromycin-susceptible staphylococci. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Clinical characteristics and persistence of bovine mastitis caused by different species of coagulase-negative staphylococci identified with API or AFLP

    DEFF Research Database (Denmark)

    Taponen, S.; Simojoki, H.; Haveri, M.;

    2006-01-01

    treatment. Mastitis due to P-lactamase-negative CNS was treated with penicillin G and that due to beta-lactamase-positive CNS with cloxacillin. Nineteen percent of the isolates were P-lactamase-positive. The bacterial cure rate for quarters treated with antimicrobials was high, 85.9%, as opposed to only 45......The coagulase-negative staphylococcal species causing mastitis in lactating cattle were identified and possible differences in the clinical characteristics or persistence of mastitis caused by different CNS were evaluated. The effect of antimicrobial treatment was also assessed. In addition, AFLP.......5% for untreated quarters. Bacterial cure rates for the most common CNS species or AFLP clusters were not statistically different. Further studies on identification of CNS species are needed....

  19. Antimicrobial susceptibility and presence of resistance genes in staphylococci from poultry

    DEFF Research Database (Denmark)

    Aarestrup, Frank Møller; Agersø, Yvonne; Ahrens, Peter

    2000-01-01

    to ciprofloxacin. Only six (7%) S. aureus isolates and one Staphylococcus saprophyticus were penicillin resistant. Resistance to sulphamethoxazole was observed among 16 (19%) of S. aureus isolates and two coagulase negative staphylococci (CNS). Twenty (24%) of the S. aureus isolates were resistant to erythromycin...... of conventional biochemical testing and 16S rDNA sequencing. The most common species were Staphylococcus aureus (83), Staphylococcus hyicus (11), Staphylococcus xylosus (9) and Staphylococcus cohnii (6). The isolates were susceptible to most antimicrobials tested. A high frequency of S. aureus (30%) was resistant...

  20. Disk Diffusion Testing for Detection of Methicillin-Resistant Staphylococci: Does Moxalactam Improve upon Cefoxitin?

    Science.gov (United States)

    Bonjean, Marie; Hodille, Elisabeth; Dumitrescu, Oana; Dupieux, Céline; Nkoud Mongo, Christina; Allam, Camille; Beghin, Mathilde; Paris, Mickael; Borrel, Ophelie; Chardon, Hubert; Laurent, Fréderic; Rasigade, Jean-Philippe; Lina, Gerard

    2016-12-01

    Disk diffusion testing is widely used to detect methicillin resistance in staphylococci, and cefoxitin is currently considered the best marker for mecA-mediated methicillin resistance. In low-inoculum diffusion testing (colony suspension at 10(6) CFU/ml), the addition of moxalactam in combination with cefoxitin has been reported to improve on cefoxitin alone for the detection of methicillin-heteroresistant staphylococci. However, moxalactam is absent from EUCAST and CLSI guidelines, which use high-inoculum diffusion testing (colony suspension at 10(8) CFU/ml), calling into question the potential interest of including moxalactam in their recommendations. The inhibition zone diameters of cefoxitin and moxalactam, alone and in combination, were evaluated for concordance with mecA and mecC positivity in a large collection of clinical Staphylococcus isolates (611 Staphylococcus aureus, Staphylococcus lugdunensis, and Staphylococcus saprophyticus isolates and 307 coagulase-negative staphylococci other than S. lugdunensis and S. saprophyticus isolates, of which 22% and 53% were mecA-positive, respectively) and in 25 mecC-positive S. aureus isolates using high-inoculum diffusion testing. Receiver operating characteristic, sensitivity, and specificity analyses indicated that the detection of mecA- and mecC-positive and negative isolates did not improve with moxalactam, either alone or in combination with cefoxitin, compared to cefoxitin alone. These findings were similar in both the S. aureus/S. lugdunensis/S. saprophyticus group and in the coagulase-negative staphylococci group. Our results do not support the use of moxalactam as an additional marker of methicillin resistance when testing with high-inoculum disk diffusion. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  1. Transmission dynamics and resistance in staphylococci

    NARCIS (Netherlands)

    Hetem, D.J.

    2015-01-01

    This thesis will focus on nosocomial transmission and resistance of S. aureus and CoNS. After the general introduction on S. aureus and coagulase-negative staphylococci, part II focuses on the nosocomial transmission capacity of different MRSA clones in the hospital setting. In chapter 2 the nosocom

  2. Transmission dynamics and resistance in staphylococci

    NARCIS (Netherlands)

    Hetem, D.J.

    2015-01-01

    This thesis will focus on nosocomial transmission and resistance of S. aureus and CoNS. After the general introduction on S. aureus and coagulase-negative staphylococci, part II focuses on the nosocomial transmission capacity of different MRSA clones in the hospital setting. In chapter 2 the

  3. Staphylococci in Competition1

    Science.gov (United States)

    Peterson, A. C.; Black, J. J.; Gunderson, M. F.

    1964-01-01

    Foods containing large amounts of carbohydrate have frequently been involved in staphylococcal food poisoning. Custard has been considered to be a highly favorable culture medium for staphylococci; however, it may be a selective medium rather than an ideal one. The influence of dextrose, lactose, and sucrose in varying amounts from 0.25 to 18%, and of starch, on the growth of staphylococci in mixed populations with saprophytes was determined. The inhibitory effect of the sugars was much greater on the saprophyte population than on the staphylococci. Of the three sugars, sucrose was most inhibitory to the saprophytes. It greatly decreased their lag periods as the concentration of sugar increased. Dextrose was the least inhibitory; in fact, 0.5% dextrose gave considerable stimulus to saprophyte growth. This sharply repressed staphylococcal growth. Lactose occupied an intermediate position. Rapid onset of the death phase of the staphylococci was observed in all increased sugar concentrations and seemed to be a pH effect rather than a result of competition. Sucrose exerted an inhibitory effect on the growth of saprophytes at and above room temperature. In the presence of 2.5% corn starch, staphylococcal growth in mixed cultures was slightly inhibited, while the death phase was sharply accelerated. Thus, carbohydrates exert their influence on staphylococcal growth in mixed cultures through their effect on the saprophytes by decreasing or increasing competition. PMID:14106944

  4. In vitro activity of ceftaroline and comparator agents against Gram-positive and Gram-negative clinical isolates from cancer patients.

    Science.gov (United States)

    Rolston, Kenneth V I; Jamal, Mohamed A; Nesher, Lior; Shelburne, Samuel A; Raad, Issam; Prince, Randall A

    2017-04-01

    Bacterial infections are common in cancer patients. Ceftaroline (CFT) is a broad-spectrum cephalosporin with activity against most Gram-positive organisms (GPOs) and many Gram-negative organisms. In this study, the in vitro activity of CFT was compared with vancomycin (VAN), daptomycin (DAP), linezolid (LZD), trimethoprim/sulphamethoxazole (SXT) and tigecycline (TIG) against bacteria (predominantly blood culture isolates) isolated from cancer patients in 2014 and 2015. CFT was active against methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA), methicillin-susceptible coagulase-negative staphylococci (MS-CoNS) and methicillin-resistant coagulase-negative staphylococci (MR-CoNS) with MIC90 values (minimum inhibitory concentration that inhibited 90% of the isolates) of 0.25, 2.0, 0.12 and 0.5 mg/L, respectively. MIC90 values for other GPOs were: Bacillus spp., >8.0 mg/L; Corynebacterium spp., 2.0 mg/L; Micrococcus spp., Gram-negative bacilli such as Enterobacter cloacae, Escherichia coli, Klebsiella spp., Proteus mirabilis and Serratia spp. Copyright © 2017 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.

  5. Performance of Variable Negative Stiffness MRE Vibration Isolation System

    Directory of Open Access Journals (Sweden)

    Run-pu Li

    2015-01-01

    Full Text Available Magnetorheological elastomer (MRE vibration isolation devices can improve a system’s vibration response via adjustable stiffness and damping under different magnetic fields. Combined with negative stiffness design, these MRE devices can reduce a system’s stiffness and improve the vibration control effect significantly. This paper develops a variable negative stiffness MRE isolation device by combining an improved separable iron core with laminated MREs. The relationship between the negative stiffness and the performance of the device is obtained by mathematical transformation. Its vibration response under simple harmonic excitation at small amplitude and the impact of the volume fraction of soft magnetic particles on the isolation system are also analyzed. The results show that the negative stiffness produced by the magnetic force is a major factor affecting the capacity of the isolation system. Compared to devices of the same size, the isolation system equipped with low-particle volume fraction MREs demonstrates better performance.

  6. Speciation and Biofilm Production of Coagulase Negative Staphylococcal Isolates from Clinically Significant Specimens and their Antibiogram

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    S. S. Vijayasri Badampudi

    2016-04-01

    Full Text Available Background: Coagulase Negative Staphylococci (CONS are increasingly recognized as significant nosocomial pathogens. Their ability of biofilm formation and multiple drug resistance are causing serious human infections. Aim and Objectives: To isolate, identify, speciate clinically significant CONS from various specimens, to study antibiotic resistance pattern and biofilm production. Material and Methods: Specimens were collected aseptically, processed and identified upto the species level by a simple scheme of tests urease, novobiocin resistance, mannose and mannitol fermentation, ornithine decarboxylase. Antibiotic sensitivity was done with special reference to methicillin resistance. Biofilm formation was detected by Congo Red Agar (CRA method and Tube Method (TM. Results: Study groupOf 100 isolates majority were pus (40, followed by urine (28, blood (16, CSF (5, body fluids (4 and catheter tips and implants (7. The most common species isolated was S. epidermidis (40% followed by S. haemolyticus (26%, S. saprophyticus (15%, S. schleiferi (13%, S. simulans (2%, S. cohnii (2% and S. warneri and S. capitis each 1%. Resistance to penicillin was 91% followed by ampicillin (79%, cotrimoxazole (67%. Methicillin resistance was 72%. Biofilm producers were 69% by CRAmethod and 33% by TM with majority species S. epidermidis (82.5%- CRA and 55%-TM. Biofilm production was significantly associated with MRCONS (p value 0.0036. Control group-Of 30 isolates were S. epidermidis 66.6% followed by S. haemolyticus (16.66%. Biofilm producers were 53.33% by CRA method and 26.65% by TM with majority species S. epidermidis (65%-CRA and 30%-TM.Methicillin resistance was 26.6%. Conclusion: Clinical significance of CONS is increasing day by day, so there is a need for accurate identification to species level and their antibiogram to avoid multidrug resistance. Biofilm producing CONS species pose a risk and CRA method for screening biofilm can be used in all conventional

  7. Prevalence of enterotoxin-encoding genes and antimicrobial resistance in coagulase-negative and coagulase-positive Staphylococcus isolates from black pudding

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    Tiane Martin de Moura

    2012-10-01

    Full Text Available INTRODUCTION: Staphylococcal species are pathogens that are responsible for outbreaks of foodborne diseases. The aim of this study was to investigate the prevalence of enterotoxin-genes and the antimicrobial resistance profile in staphylococcus coagulase-negative (CoNS and coagulasepositive (CoPS isolates from black pudding in southern Brazil. METHODS: Two hundred typical and atypical colonies from Baird-Parker agar were inoculated on mannitol salt agar. Eighty-two mannitol-positive staphylococci were submitted to conventional biochemical tests and antimicrobial susceptibility profiling. The presence of coagulase (coa and enterotoxin (se genes was investigated by polymerase chain reaction. RESULTS: The isolates were divided into 2 groups: 75.6% (62/82 were CoNS and 24.4% (20/82 were CoPS. The biochemical tests identified 9 species, of which Staphylococcus saprophyticus (37.8% and Staphylococcus carnosus (15.9% were the most prevalent. Antimicrobial susceptibility tests showed resistance phenotypes to antibiotics widely administered in humans, such as gentamicin, tetracycline, chloramphenicol, and erythromycin. The coa gene was detected in 19.5% (16/82 of the strains and 4 polymorphic DNA fragments were observed. Five CoNS isolates carrying the coa gene were submitted for 16S rRNA sequencing and 3 showed similarity with CoNS. Forty strains were positive for at least 1 enterotoxin-encoding gene, the genes most frequently detected were sea (28.6% and seb (27.5%. CONCLUSIONS: The presence of antimicrobial resistant and enterotoxin-encoding genes in staphylococci isolates from black pudding indicated that this fermented food may represent a potential health risk, since staphylococci present in food could cause foodborne diseases or be a possible route for the transfer of antimicrobial resistance to humans.

  8. Multicenter Evaluation of the Staphylococcus QuickFISH Method for Simultaneous Identification of Staphylococcus aureus and Coagulase-Negative Staphylococci Directly from Blood Culture Bottles in Less than 30 Minutes

    Science.gov (United States)

    Deck, Melissa K.; Anderson, Erica S.; Buckner, Rebecca J.; Colasante, Georgia; Coull, James M.; Crystal, Benjamin; Latta, Phyllis Della; Fuchs, Martin; Fuller, Deanna; Harris, Will; Hazen, Kevin; Klimas, Lisa L.; Lindao, Daniel; Meltzer, Michelle C.; Morgan, Margie; Shepard, Janeen; Stevens, Sharon; Wu, Fann

    2012-01-01

    A novel rapid peptide nucleic acid fluorescence in situ hybridization (FISH) method, Staphylococcus QuickFISH, for the direct detection of Staphylococcus species from positive blood culture bottles was evaluated in a multicenter clinical study. The method utilizes a microscope slide with predeposited positive- and negative-control organisms and a self-reporting 15-min hybridization step, which eliminates the need for a wash step. Five clinical laboratories tested 722 positive blood culture bottles containing Gram-positive cocci in clusters. The sensitivities for detection of Staphylococcus aureus and coagulase-negative staphylococci (CoNS) were 99.5% (217/218) and 98.8% (487/493), respectively, and the combined specificity of the assay was 89.5% (17/19). The combined positive and negative predictive values of the assay were 99.7% (696/698) and 70.8% (17/24), respectively. Studies were also performed on spiked cultures to establish the specificity and performance sensitivity of the method. Staphylococcus QuickFISH has a turnaround time (TAT) of <30 min and a hands-on time (HOT) of <5 min. The ease and speed of the method have the potential to improve the accuracy of therapeutic intervention by providing S. aureus/CoNS identification simultaneously with Gram stain results. PMID:22493336

  9. Multicenter evaluation of the Staphylococcus QuickFISH method for simultaneous identification of Staphylococcus aureus and coagulase-negative staphylococci directly from blood culture bottles in less than 30 minutes.

    Science.gov (United States)

    Deck, Melissa K; Anderson, Erica S; Buckner, Rebecca J; Colasante, Georgia; Coull, James M; Crystal, Benjamin; Della Latta, Phyllis; Fuchs, Martin; Fuller, Deanna; Harris, Will; Hazen, Kevin; Klimas, Lisa L; Lindao, Daniel; Meltzer, Michelle C; Morgan, Margie; Shepard, Janeen; Stevens, Sharon; Wu, Fann; Fiandaca, Mark J

    2012-06-01

    A novel rapid peptide nucleic acid fluorescence in situ hybridization (FISH) method, Staphylococcus QuickFISH, for the direct detection of Staphylococcus species from positive blood culture bottles was evaluated in a multicenter clinical study. The method utilizes a microscope slide with predeposited positive- and negative-control organisms and a self-reporting 15-min hybridization step, which eliminates the need for a wash step. Five clinical laboratories tested 722 positive blood culture bottles containing gram-positive cocci in clusters. The sensitivities for detection of Staphylococcus aureus and coagulase-negative staphylococci (CoNS) were 99.5% (217/218) and 98.8% (487/493), respectively, and the combined specificity of the assay was 89.5% (17/19). The combined positive and negative predictive values of the assay were 99.7% (696/698) and 70.8% (17/24), respectively. Studies were also performed on spiked cultures to establish the specificity and performance sensitivity of the method. Staphylococcus QuickFISH has a turnaround time (TAT) of <30 min and a hands-on time (HOT) of <5 min. The ease and speed of the method have the potential to improve the accuracy of therapeutic intervention by providing S. aureus/CoNS identification simultaneously with Gram stain results.

  10. Fluoroquinolone susceptibilities to methicillin-resistant and susceptible coagulase-negative Staphylococcus isolated from eye infection Suscetibilidade dos Staphylococcus coagulase negativo meticilina-resistentes e suscetíveis isolados em infecções oculares

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    Adália Dias Dourado Oliveira

    2007-03-01

    Full Text Available PURPOSE: To evaluate the fluoroquinolone susceptibilities of ocular isolate coagulase-negative staphylococci (CoNS, identified at the Microbiology Laboratory - UNIFESP. DESIGN: Experimental laboratory investigation. METHODS: The minimum inhibitory concentrations (MICs of 21 strains of methicillin-resistant coagulase-negative staphylococci (MRCoNS and 22 methicillin-sensitive coagulase-negative staphylococci (MSCoNS to ciprofloxacin, ofloxacin, gatifloxacin and moxifloxacin were determined, using the E-test method standardized by the Clinical and Laboratory Standards Institute (CLSI/NCCLS. RESULTS: The MIC90s (µg/ml for the second generation of tested fluoroquinolones were higher than the fourth generation, especially for the methicillin-resistant coagulase-negative staphylococci group. CONCLUSION: Our results indicate that methicillin-sensitive coagulase-negative staphylococci are more susceptible to quinolones than are methicillin-resistant coagulase-negative staphylococci and that fourth generation fluoroquinolones appear to be more potent, affecting even coagulase-negative staphylococcal strains resistant to second generation fluoroquinolones.OBJETIVOS: Avaliar a suscetibilidade a fluorquinolonas dos Staphylococcus coagulase-negativo (SCoN identificados no Laboratório de Microbiologia Ocular da Unifesp. MÉTODOS: Foi determinada a concentração inibitória mínima de 21 cepas de SCoN meticilina-resistentes e 22 meticilina-sensíveis para ciprofloxacina, ofloxacina, gatifloxacina e moxifloxacina, utilizando o E-test estandartizado pelo CLSI/NCCLS. RESULTADOS: Os MIC90 (µg/ml de 43 SCoN isolados para fluorquinolonas de segunda geração foram maiores do que os de quarta geração, principalmente para o grupo dos meticilina-resistentes. CONCLUSÃO: Nossos resultados indicam que Staphylococcus coagulase-negativo meticilina-sensíveis são mais suscetíveis às quinolonas do que os Staphylococcus coagulase-negativo meticilina

  11. Identification of Variable Traits among the Methicillin Resistant and Sensitive Coagulase Negative Staphylococci in Milk Samples from Mastitic Cows in India

    National Research Council Canada - National Science Library

    Sudipta Mahato; Hiral U. Mistry; Shalini Chakraborty; Paresh Sharma; R. Saravanan; Vasundhra Bhandari

    2017-01-01

    ... determinants, virulence and toxin genes present in the CoNS causing clinical mastitis. We obtained 62 CoNS isolates from 167 mastitic milk samples collected from three different states of India...

  12. Nasal carriage of methicillin resistant staphylococci

    DEFF Research Database (Denmark)

    Faria, Nuno A; Conceição, Teresa; Miragaia, Maria;

    2014-01-01

    Coagulase-negative staphylococci (CoNS) are believed to function as reservoirs, as well as possible sources of staphylococcal chromosome cassette mec (SCCmec) to Staphylococcus aureus, but the frequency, preferred partners, and factors promoting SCCmec transfer are not known. Such postulated....... A total of 54 patients (80%) were colonized with staphylococci that included nine different species identified by internal transcribed spacer PCR (ITS-PCR) and 16S RNA sequencing. The highest rates of colonization were found for S. epidermidis (58%) and S. aureus (39%). Methicillin resistance was present...... in S. aureus (53%), S. epidermidis (53%), S. haemolyticus (33%), and S. hominis (62%). Genetic backgrounds were characterized by spa typing for S. aureus and by pulsed-field gel electrophoresis for CoNS. SCCmec typing showed that SCCmec type IV (2B) was the most common in the entire collection (65...

  13. 凝固酶阴性的葡萄球菌与早产儿脑损伤的关系%The relationship between coagulase-negative staphylococci and preterm brain injury

    Institute of Scientific and Technical Information of China (English)

    乔丽丽

    2012-01-01

    Coagulase-negative staphylococci,especially staphylococcus epidermidis (SE) is on the surface of skin and mucous membrane of human,and because of its high frequency,especially in hospital,it is thought to be an important opportunistic pathogen.SE is the common cause of preterm infection,and it has relationship with immature brain.Epidemiology supports the relationship among preterm,infection and brain injury.SE could lead to brain injury through Toll-like receptor mediated inflammation.%凝固酶阴性的葡萄球菌,尤其是表皮葡萄球菌(staphylococcus epidermidis,SE)存在于健康人的皮肤和黏膜上,它在医院内有高感染率,是重要的条件致病菌.SE是早产儿感染的最常见原因,这与早产儿大脑发育不成熟有关.流行病学研究支持早产儿、细菌感染和脑损伤之间存在相关性,SE可能是通过Toll样受体介导炎症反应而引起早产儿脑损伤.

  14. Genome Sequences of Nine Gram-Negative Vaginal Bacterial Isolates

    Science.gov (United States)

    Deitzler, Grace E.; Ruiz, Maria J.; Lu, Wendy; Weimer, Cory; Park, SoEun; Robinson, Lloyd S.; Hallsworth-Pepin, Kymberlie; Wollam, Aye; Mitreva, Makedonka

    2016-01-01

    The vagina is home to a wide variety of bacteria that have great potential to impact human health. Here, we announce reference strains (now available through BEI Resources) and draft genome sequences for 9 Gram-negative vaginal isolates from the taxa Citrobacter, Klebsiella, Fusobacterium, Proteus, and Prevotella. PMID:27688330

  15. Plating isolation of various catalase-negative microorganisms from soil

    Science.gov (United States)

    Labeda, D. P.; Hunt, C. M.; Casida, L. E., Jr.

    1974-01-01

    A unique plating procedure was developed that allows isolation, but not enumeration, of representatives of the catalase-negative soil microflora. The numbers recovered, however, are low as compared to the numbers recovered when the modified dilution-to-extinction isolation procedure is used. The latter procedure provides prolonged inoculation in sealed tubes containing a nutritionally rich broth medium over small submerged agar slants. In contrast, the plating procedure utilizes nutritionally minimal media and the shorter incubations mandated by the inherent problems associated with plating.

  16. Plating isolation of various catalase-negative microorganisms from soil

    Science.gov (United States)

    Labeda, D. P.; Hunt, C. M.; Casida, L. E., Jr.

    1974-01-01

    A unique plating procedure was developed that allows isolation, but not enumeration, of representatives of the catalase-negative soil microflora. The numbers recovered, however, are low as compared to the numbers recovered when the modified dilution-to-extinction isolation procedure is used. The latter procedure provides prolonged inoculation in sealed tubes containing a nutritionally rich broth medium over small submerged agar slants. In contrast, the plating procedure utilizes nutritionally minimal media and the shorter incubations mandated by the inherent problems associated with plating.

  17. Antibiotic surgical prophylaxis increases nasal carriage of antibiotic-resistant staphylococci.

    Science.gov (United States)

    McMurray, Claire L; Hardy, Katherine J; Verlander, Neville Q; Hawkey, Peter M

    2015-12-01

    Staphylococci are a significant cause of hospital-acquired infection. Nasal carriage of Staphylococcus aureus is an important risk factor for infection in surgical patients and coagulase-negative staphylococci (CNS) are a major cause of prosthetic joint infections. The impact that antibiotic surgical prophylaxis has on the nasal carriage of staphylococci has not been studied. Daily nasal swabs were taken from 63 patients who received antibiotic surgical prophylaxis and 16 patients who received no antibiotics. Total aerobic bacterial count, S. aureus and CNS were enumerated by culture from nasal swabs. Representative isolates were typed by staphylococcal interspersed repeat units (SIRU) typing and PFGE, and MICs to nine antibiotics were determined. After antibiotic administration, there was a reduction in S. aureus counts (median - 2.3 log(10)c.f.u. ml(- 1)) in 64.0 % of S. aureus carriers, compared with only a 0.89 log(10)c.f.u. ml(- 1) reduction in 75.0 % of S. aureus carriers who did not receive antibiotics. A greater increase in the nasal carriage rate of meticillin-resistant CNS was observed after antibiotic surgical prophylaxis compared with hospitalization alone, with increases of 16.4 and 4.6 %, respectively. Antibiotic-resistant S. epidermidis carriage rate increased by 16.6 % after antibiotic administration compared with 7.5 % with hospitalization alone. Antibiotic surgical prophylaxis impacts the nasal carriage of both S. aureus and CNS.

  18. Characterization of methicillin-susceptible and -resistant staphylococci in the clinical setting: a multicentre study in Nigeria

    Directory of Open Access Journals (Sweden)

    Shittu Adebayo

    2012-11-01

    Full Text Available Abstract Background The staphylococci are implicated in a variety of human infections; however, many clinical microbiology laboratories in Nigeria do not identify staphylococci (in particular coagulase negative staphylococci - CNS to the species level. Moreover, data from multi-centre assessment on antibiotic resistance and epidemiology of the staphylococci are not available in Nigeria. This study investigated 91 non-duplicate staphylococcal isolates obtained from the microbiology laboratories of eight hospitals in Nigeria during the period January to April 2010. Methods Identification and antibiotic susceptibility testing was performed using the VITEK 2 system, detection of resistance genes by PCR, and molecular characterization was determined by SCCmec typing, spa and multilocus sequence typing (MLST. Results All the isolates were susceptible to mupirocin, tigecycline, vancomycin and linezolid, but 72.5% of CNS and 82.3% of Staphylococcus aureus were resistant to cotrimoxazole, while multiresistance was observed in 37 of the 40 CNS isolates. Untypeable SCCmec types (ccrC/Class A mec and ccr-negative/Class C2 mec gene complex in two methicillin-resistant S. aureus (MRSA were identified. Additionally, ccr-negative/Class A mec and ccr type 4/Class C2 mec gene complex was detected in one isolate each of S. sciuri and S. haemolyticus, respectively. The S. aureus isolates were classified into 21 spa types including two new types (t8987, t9008 among the methicillin-susceptible S. aureus (MSSA isolates. Two (CC8-SCCmecnon-typeable and CC88-SCCmec IV and four (CC8-SCCmec III/IV/V; CC30-SCCmec II/III; CC88-SCCmec IV; and ST152-SCCmecnon-typeable MRSA clones were identified in Maiduguri (North-East Nigeria and South-West Nigeria, respectively. The proportion of Panton-Valentine leukocidin (PVL-positive MSSA was high (44.4% and 56.3% of these strains were associated with sequence type (ST 152. Conclusions The identification of multiresistant mecA positive

  19. 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.

  20. THERAPEUTIC IMPORTANCE OF OLDER GENERATION ANTIBIOTICS ON GRAM NEGATIVE ISOLATES

    OpenAIRE

    B.Lakshmi, M.Swarajya Lakshmi, Rohini

    2015-01-01

    Introduction: Drug resistance is a serious medical problem. Indiscriminate use of antibiotics has led to a state where multi drug resistant bacteria have become increasingly prevalent. Therefore regular surveillance of important pathogens and their resistant pattern is mandatory. Aim: To find out prevalence of organisms causing infection and their sensitivity pattern. Material and methods: 676 clinical samples were screened among which 156 Gram Negative(GN) Isolates were processed for their a...

  1. THERAPEUTIC IMPORTANCE OF OLDER GENERATION ANTIBIOTICS ON GRAM NEGATIVE ISOLATES

    OpenAIRE

    Lakshmi; Swarajya Lakshmi; Rohini,

    2015-01-01

    Drug resistance is a serious medical problem. Indiscriminate use of antibiotics has led to a state where multi-drug resistant bacteria have become increasingly prevalent. Therefore regular surveillance of important pathogens and their resistant pattern is mandatory. AIM To find out prevalence of organisms causing infection and their sensitivity pattern. MATERIAL AND METHODS 676 clinical samples were screened, among which 156 Gram Negative (GN) Isolates were ...

  2. Epidemiology of drug resistance: The case of Staphylococcus aureus and coagulase-negative staphylococci infections Epidemiología de la resistencia bacteriana: el caso de Staphylococcus aureus y las infecciones Staphylococcus coagulasa negativas

    Directory of Open Access Journals (Sweden)

    Ernesto Calderón-Jaimes

    2002-04-01

    Full Text Available Objective. To study the activity of several antibiotics against Staphylococcus spp. Material and Methods. The study included 1209 strains of Staphylococcus spp. from two institutions; Instituto Nacional de Pediatría (National Institute of Pediatrics and Hospital Infantil de México Federico Gómez (Mexico City Children's Hospital. Minimum Inhibitory Concentrations of all antibiotics were determined by the agar macrodilution technique and standard methods from the National Committee for Clinical Laboratory Standards. Results. Resistance of S. aureus was 14.2% and that of coagulase-negative staphylococci was 53.4%. The activity of different antibiotics is presented in detail. Conclusions. Surveillance of strains resistant to methicillin is necessary.Objetivo. Determinar la frecuencia de la resistencia a la meticilina y la actividad de varios antibióticos. Material y métodos. Se incluyeron 1 209 cepas de Staphylococcus spp. procedentes de pacientes del Instituto Nacional de Pediatría y del Hospital Infantil de México Federico Gómez. Se utilizó la técnica de dilución en placas con agar. El procedimiento e interpretación fueron acordes con lo establecido por el National Committee for Clinical Laboratory Standards. Resultados. La frecuencia de la resistencia de S. aureus fue de 14.2% y de 53.4% en los Staphylococcus coagulasa negativa. La actividad de otros antimicrobianos se presenta en el texto. Conclusiones. Es necesario vigilar continuamente la progresión de la resistencia de Staphylococcus spp. a la meticilina.

  3. THERAPEUTIC IMPORTANCE OF OLDER GENERATION ANTIBIOTICS ON GRAM NEGATIVE ISOLATES

    Directory of Open Access Journals (Sweden)

    Lakshmi

    2015-12-01

    Full Text Available Drug resistance is a serious medical problem. Indiscriminate use of antibiotics has led to a state where multi-drug resistant bacteria have become increasingly prevalent. Therefore regular surveillance of important pathogens and their resistant pattern is mandatory. AIM To find out prevalence of organisms causing infection and their sensitivity pattern. MATERIAL AND METHODS 676 clinical samples were screened, among which 156 Gram Negative (GN Isolates were processed for their antibiotic sensitivity profile against 12 different antibiotics. RESULTS Escherichia coli is the most common isolate of 156 gram negative isolates. Among all antibiotics, Ampicillin is least sensitive (22%. Antibiotics with good sensitivity are Imipenem, Meropenem (100%, Levofloxacin 94%, Amikacin 89%, Ciprofloxacin 79%, Gentamicin 77%. Pseudomonas is 100% sensitive to Amikacin. CONCLUSION Antibiotic resistance in our area is still moderate. It is essential to test for older generation antibiotics before deciding on higher antibiotics for treatment, which will have a tremendous impact on the treatment as well as cost effectiveness. Regular surveillance helps in implementing better therapeutic strategies.

  4. THERAPEUTIC IMPORTANCE OF OLDER GENERATION ANTIBIOTICS ON GRAM NEGATIVE ISOLATES

    Directory of Open Access Journals (Sweden)

    B.Lakshmi, M.Swarajya Lakshmi, Rohini

    2015-10-01

    Full Text Available Introduction: Drug resistance is a serious medical problem. Indiscriminate use of antibiotics has led to a state where multi drug resistant bacteria have become increasingly prevalent. Therefore regular surveillance of important pathogens and their resistant pattern is mandatory. Aim: To find out prevalence of organisms causing infection and their sensitivity pattern. Material and methods: 676 clinical samples were screened among which 156 Gram Negative(GN Isolates were processed for their antibiotic sensitivity profile against 12 different antibiotics. Results: Escherichia coli is the most common isolate of 156 gram negative isolates. Among all antibiotics, ampicillin is least sensitive (22%. Antibiotics with good sensitivity are Imipenam, Meropenam (100%, Levofloxacin 94%, Amikacin 89% Ciprofloxacin 79%, Gentamycin 77%. Pseudomonas is 100% sensitive to Amikacin. Conclusion: Antibiotic resistance in our area is still moderate. It is essential to test for older generation antibiotics before deciding on higher antibiotics for treatment which will have a tremendous impact on the treatment as well as cost effectiveness. Regular surveillance helps in implementing better therapeutic strategies.

  5. [Microbiological profiling of isolates from the neonatal unit of a third-level hospital in Bogotá, Colombia].

    Science.gov (United States)

    Cifuentes, Yolanda; Ruiz, Ariel I; Leal, Aura L; Muñoz, Liliana C; Herrera, Maria T; Jiménez, Luz M

    2005-01-01

    Investigating the prevalence and sensitivity of germs isolated from newborn in a referral hospital in Bogotá. Suggesting an empirical antibiotic treatment for neonatal infection. Cultures taken between February and December 2002 were analysed. Blood cultures were processed using BacT/ALERT (Durham, NC), urine cultures by UROCULT (Bio-Bacter) and catheter tips in thioglycollate. BBL CRYSTAL identification system (BD, Sparks, MD) was used for identifying germs. Antibiotic sensitivity was determined by disk diffusion. There were 1,097 positive aerobic and facultative aerobic germ cultures; 64.3% were Gram-positive, 30.6% Gram-negative and 4.9% were yeasts. Gram-positive germs consisted of coagulase-negative staphylococci (64.2%), enterococcus (13.8%) and coagulase-positive staphylococci (13.3%). The most frequent Gram-negatives were Klebsiella (45.2%), Eschericha coli (30.9%) and Serratia (10.1%). Staphylococcus epidermidis accounted for 64% of the coagulase-negative staphylococci. S. epidermidis susceptibility to vancomycin was 100%. Coagulase-negative staphylococci susceptibility to rifampin and amikacin was 59% and 67.4% (respectively). Coagulase-negative staphylococci resistance to beta-lactams was 86.4% (95% CI: 82.3-89.9). Coagulase-positive staphylococci sensitivity to vancomycin was 100%. Gram-negative susceptibility to imipenem was 98.1% (95% CI: 89.9-99.9), 78.1% to gentamicin (95% CI: 64.9-88.2) and 46.6% to amikacin (95% CI: 28.3-65.7). There was high coagulase-negative staphylococci prevalence in neonatal infection (particularly S. epidermidis). All S. epidermidis and coagulase-positive staphylococci were sensitive to vancomycin. There was increasing coagulase-negative staphylococci and Gram-negative resistance to oxacillin and amikacin, respectively.

  6. Biofilm production by clinical staphylococci strains from canine otitis

    Science.gov (United States)

    Moreira, Camila Alencar; de Oliveira, Lis Christina; Mendes, Marina Silveira; Santiago, Thiago de Melo; Barros, Eduardo Bedê; de Carvalho, Cibele Barreto Mano

    2012-01-01

    This study determined the species of 54 staphylococci isolates from canine otitis and their ability to produce biofilm through the Congo red agar method, confirmed by scanning electron microscopy. The most frequently identified species were S. intermedius and S. simulans. Results showed that 30% of the strains were biofilm producers. PMID:24031841

  7. Distribution of classical enterotoxin genes in staphylococci from milk of cows with- and without mastitis and the cowshed environment.

    Science.gov (United States)

    Piechota, M; Kot, B; Zdunek, E; Mitrus, J; Wicha, J; Wolska, M K; Sachanowicz, K

    2014-01-01

    The aim of this study was to analyze by PCR 185 isolates of Staphylococcus from milk of cows with- and without mastitis and from the cowsheds environment for their potential ability to produce five classical staphylococcal enterotoxins. Among S. aureus isolates 8 (32%) carried enterotoxin genes and only 2 of them had more than one gene. The enterotoxin genes were detected in 22 (13.7%) coagulase-negative staphylococci (CNS) isolates, among them in 9 (11.4%) isolates of S. xylosus, 5 (16.7%) S. sciuri, 3 (10.3%) S. epidermidis and in 5 (22.7%) Staphylococcus spp. In some CNS 2 or 3 genes were detected simultaneously. Among the investigated enterotoxin genes, sec was the most prevalent (70%). The genes encoding enterotoxin B and D were detected in 5 (16.7%) and 6 (20%) isolates, respectively. The lowest number of isolates had sea and see genes. The genes encoding enterotoxins were often identified in staphylococci from milk of cows with mastitis (73.4% of detected genes), while only 6 (20%) isolates from milk of cows without mastitis and 2 (6.6%) isolates from cowshed environment were positive for enterotoxin genes. The results showed that CNS from bovine milk, like S. aureus, carried enterotoxin genes and may pose a risk for public health.

  8. The warmer the weather, the more gram-negative bacteria - impact of temperature on clinical isolates in intensive care units.

    Directory of Open Access Journals (Sweden)

    Frank Schwab

    Full Text Available BACKGROUND: We investigated the relationship between average monthly temperature and the most common clinical pathogens causing infections in intensive care patients. METHODS: A prospective unit-based study in 73 German intensive care units located in 41 different hospitals and 31 different cities with total 188,949 pathogen isolates (102,377 Gram-positives and 86,572 Gram-negatives from 2001 to 2012. We estimated the relationship between the number of clinical pathogens per month and the average temperature in the month of isolation and in the month prior to isolation while adjusting for confounders and long-term trends using time series analysis. Adjusted incidence rate ratios for temperature parameters were estimated based on generalized estimating equation models which account for clustering effects. RESULTS: The incidence density of Gram-negative pathogens was 15% (IRR 1.15, 95%CI 1.10-1.21 higher at temperatures ≥ 20°C than at temperatures below 5°C. E. cloacae occurred 43% (IRR=1.43; 95%CI 1.31-1.56 more frequently at high temperatures, A. baumannii 37% (IRR=1.37; 95%CI 1.11-1.69, S. maltophilia 32% (IRR=1.32; 95%CI 1.12-1.57, K. pneumoniae 26% (IRR=1.26; 95%CI 1.13-1.39, Citrobacter spp. 19% (IRR=1.19; 95%CI 0.99-1.44 and coagulase-negative staphylococci 13% (IRR=1.13; 95%CI 1.04-1.22. By contrast, S. pneumoniae 35% (IRR=0.65; 95%CI 0.50-0.84 less frequently isolated at high temperatures. For each 5°C increase, we observed a 3% (IRR=1.03; 95%CI 1.02-1.04 increase of Gram-negative pathogens. This increase was highest for A. baumannii with 8% (IRR=1.08; 95%CI 1.05-1.12 followed by K. pneumoniae, Citrobacter spp. and E. cloacae with 7%. CONCLUSION: Clinical pathogens vary by incidence density with temperature. Significant higher incidence densities of Gram-negative pathogens were observed during summer whereas S. pneumoniae peaked in winter. There is increasing evidence that different seasonality due to physiologic changes underlies

  9. Nationwide German Multicenter Study on Prevalence of Antibiotic Resistance in Staphylococcal Bloodstream Isolates and Comparative In Vitro Activities of Quinupristin-Dalfopristin

    OpenAIRE

    Von Eiff, Christof; Reinert, Ralf René; Kresken, Michael; Brauers, Johannes; Hafner, Dieter; Peters, Georg

    2000-01-01

    Antibiotic-resistant gram-positive bacteria have become an increasing problem in the last two decades. In order to evaluate the prevalence of antibiotic resistance in staphylococcal bloodstream isolates in Germany, 2,042 staphylococci collected in 21 tertiary-care hospitals were investigated during a 3-year period (March 1996 to March 1999). Altogether, 1,448 S. aureus isolates and 594 coagulase-negative staphylococci (CoNS) that comprised 13 different species were included. Furthermore, the ...

  10. Staphylococcus chromogenes, a Coagulase-Negative Staphylococcus Species That Can Clot Plasma.

    Science.gov (United States)

    Dos Santos, Danielle Cabral; Lange, Carla Christine; Avellar-Costa, Pedro; Dos Santos, Katia Regina Netto; Brito, Maria Aparecida Vasconcelos Paiva; Giambiagi-deMarval, Marcia

    2016-05-01

    Staphylococcus chromogenes is one of the main coagulase-negative staphylococci isolated from mastitis of dairy cows. We describe S. chromogenes isolates that can clot plasma. Since the main pathogen causing mastitis is coagulase-positive Staphylococcus aureus, the coagulase-positive phenotype of S. chromogenes described here can easily lead to misidentification. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  11. Coagulase-negative staphylococci as reservoirs of genes facilitating MRSA infection: Staphylococcal commensal species such as Staphylococcus epidermidis are being recognized as important sources of genes promoting MRSA colonization and virulence.

    Science.gov (United States)

    Otto, Michael

    2013-01-01

    Recent research has suggested that Staphylococcus epidermidis is a reservoir of genes that, after horizontal transfer, facilitate the potential of Staphylococcus aureus to colonize, survive during infection, or resist antibiotic treatment, traits that are notably manifest in methicillin-resistant S. aureus (MRSA). S. aureus is a dangerous human pathogen and notorious for acquiring antibiotic resistance. MRSA in particular is one of the most frequent causes of morbidity and death in hospitalized patients. S. aureus is an extremely versatile pathogen with a multitude of mechanisms to cause disease and circumvent immune defenses. In contrast, most other staphylococci, such as S. epidermidis, are commonly benign commensals and only occasionally cause disease. Recent findings highlight the key importance of efforts to better understand how genes of staphylococci other than S. aureus contribute to survival in the human host, how they are transferred to S. aureus, and why this exchange appears to be uni-directional.

  12. The Staphylococci Phages Family: An Overview

    Directory of Open Access Journals (Sweden)

    Laurence Van Melderen

    2012-11-01

    Full Text Available Due to their crucial role in pathogenesis and virulence, phages of Staphylococcus aureus have been extensively studied. Most of them encode and disseminate potent staphylococcal virulence factors. In addition, their movements contribute to the extraordinary versatility and adaptability of this prominent pathogen by improving genome plasticity. In addition to S. aureus, phages from coagulase-negative Staphylococci (CoNS are gaining increasing interest. Some of these species, such as S. epidermidis, cause nosocomial infections and are therefore problematic for public health. This review provides an overview of the staphylococcal phages family extended to CoNS phages. At the morphological level, all these phages characterized so far belong to the Caudovirales order and are mainly temperate Siphoviridae. At the molecular level, comparative genomics revealed an extensive mosaicism, with genes organized into functional modules that are frequently exchanged between phages. Evolutionary relationships within this family, as well as with other families, have been highlighted. All these aspects are of crucial importance for our understanding of evolution and emergence of pathogens among bacterial species such as Staphylococci.

  13. Occurrence, species distribution, antimicrobial resistance and clonality of methicillin- and erythromycin-resistant staphylococci in the nasal cavity of domestic animals

    DEFF Research Database (Denmark)

    Bagcigil, Funda A.; Moodley, Arshnee; Baptiste, Keith E.

    2007-01-01

    , horses, pigs, and cattle in Denmark. Nasal swabs were collected from a total of 400 animals, including 100 individuals of each species. Methicillin and erythromycin-resistant staphylococci were isolated on selective media, identified by 16S rDNA sequencing, and typed by pulsed field gel electrophoresis...... (PFGE). Methicillin-resistant coagulase-negative staphylococci (MRCoNS) harbouring mecA were isolated from horses (50%) and dogs (13%), but not from food animals. The species identified were S. haemolyticus (n = 21), S. vitulinus (n = 19), S. sciuri (n = 13), S. epidermidis (n = 8), and S. warneri (n...... = 2). mecA-mediated methicillin resistance in S. vitulinus was described for the first time. Methicillin-resistant S. aureus was not detected. PFGE analysis revealed the presence of specific MRCoNS clones in samples originating from the same veterinary hospital or equine farm. Erythromycin-resistant S...

  14. Value of number of bottles of positive blood culture in diagnosis of bloodstream infections caused by coagulase-negative Staphylococci%血培养报阳瓶数对凝固酶阴性葡萄球菌血流感染鉴别诊断的价值

    Institute of Scientific and Technical Information of China (English)

    黄声雷; 胡必杰; 谢红梅; 周春妹; 马艳

    2014-01-01

    目的:探讨血培养报阳瓶数在鉴别凝固酶阴性葡萄球菌(CNS)血流感染中的应用价值。方法采用回顾性研究方法,以2012年1月-2013年12月复旦大学附属中山医院血培养结果为CNS的住院患者370例为研究对象,收集菌株鉴定和药敏试验结果,并根据标准判断阳性培养是否血流感染或污染,借此评价血培养报阳瓶数在CNS血流感染鉴别诊断中的价值。结果 CNS阳性血培养中78例系CNS引起的菌血症,占21.1%,212例则由CNS污染造成,占57.3%,80例临床意义不明确,占21.6%;以双套4瓶血培养中的CNS阳性瓶数进行判断,1瓶阳性判断为血流感染30例,占38.5%、2瓶12例占15.4%、3瓶20例占25.6%、4瓶16例占20.5%;根据双套血培养中不同阳性瓶数来判断由CNS引起的菌血症的阳性预期值分别为1瓶16.3%、2瓶30.0%、3瓶50.0%和4瓶61.5%。结论虽然血培养报阳性瓶数单独作为鉴别诊断CNS血流感染和血培养污染的指标具有不稳定性,但与患者的临床其他指标相结合,其对早期识别阳性血培养为污染菌或感染菌,具有参考价值。%OBJECTIVE To explore the value of number of bottles of positive blood culture in identification of blood-stream infections caused by coagulase-negative Staphylococci .METHODS By means of retrospective survey ,totally 370 patients who were hospitalized the Zhongshan Hospital Affiliated to Fudan University from Jan 2012 to Dec 2013 were enrolled in the study ,the coagulase-negative Staphylococci strains were cultured from the blood speci-mens obtained from the subjects ,then the results of the identification of strains and the drug susceptibility testing were collected ,the criteria were referred to determine the bloodstream infections or contamination in the positive culture ,and the value of the number of bottles of positive blood culture in the diagnosis of coagulase-negative

  15. Antimicrobial resistance and genotypes of staphylococci from bovine milk and the cowshed environment.

    Science.gov (United States)

    Kot, B; Piechota, M; Antos-Bielska, M; Zdunek, E; Wolska, K M; Binek, T; Olszewska, J; Guliński, P; Trafny, E A

    2012-01-01

    Investigation of antimicrobial resistance and genetic relatedness of staphylococci from milk of cows with mastitis and cowshed environment was the aim of this study. Antimicrobial resistance against 14 antimicrobials were determined by using a disc diffusion method. Genetic similarity between the most frequently isolated species was analysed by PFGE (pulsed-field gel electrophoresis). Haemolytic activity, DNase, protease and esterase production was also investigated. Coagulase-negative Staphylococcus species were isolated from 30.8% of milk samples from cows with mastitis. The most frequently isolated species was Staphylococcus xylosus and yield of these organisms was significantly associated with milk of mastitis cows. S. epidermidis was a predominant penicillin-resistant species. High frequency of resistance to lincomycin was observed among isolates of S. sciuri (54.2%) and S. xylosus (25.9%) from cows with mastitis. PFGE (pulsed-field gel electrophoresis) analysis of 29 Staphylococcus aureus isolates showed the presence of 17 PFGE pulsotypes. Isolates of S. sciuri (n = 36) had unique PFGE patterns. Some S. xylosus isolates from milk and milker's hands had the same PFGE pulsotypes, and this observation could indicate that dairyman may be a potential source of the infection. The pulsotype of each of the remaining isolates of S. xylosus suggested that they might have come from common environmental sources; however, these isolates differed in antibiotic resistance pattern or virulence traits. Therefore, knowledge about antibiotic sensitivity pattern and virulence factors of a CNS isolate, besides its genotype, may be informative in tracking the source of the infection.

  16. Occurrence and characterization of methicillin-resistant staphylococci from bovine mastitis milk samples in Finland.

    Science.gov (United States)

    Gindonis, Veera; Taponen, Suvi; Myllyniemi, Anna-Liisa; Pyörälä, Satu; Nykäsenoja, Suvi; Salmenlinna, Saara; Lindholm, Laura; Rantala, Merja

    2013-08-28

    Methicillin-resistant staphylococci (MRS) are increasingly being isolated in bovine mastitis. The aim of our study was to evaluate the occurrence of MRS in Finnish mastitis milk samples and characterize the MRS isolates using molecular methods. Methicillin-resistant S. aureus (MRSA) was a rare finding in bovine mastitis in Finland. Only two out of 135 (1.5%) S. aureus isolates were positive for mec genes. One of these carried mecA and was of spa type t172, SCCmec type IV and ST375, and the other harboured mecC, being spa type t3256, and ST130. MRSA ST375 is common among human MRSA isolates in Finland, but this is the first report in the country of bovine mecC MRSA. In coagulase-negative staphylococci (CoNS) originating from bovine mastitis, methicillin resistance was more common. In the two CoNS collections studied, 5.2% (17/324) and 1.8% (2/110) of the isolates were mecA positive. Eighteen of these were methicillin-resistant S. epidermidis (MRSE), which were divided into 6 separate PFGE clusters. One pulsotype was detected in different parts of the country, indicating clonal spread. Most MRSE (13/18) were of SCCmec type IV, one was of type V and four were non-typeable. Comparison with a human staphylococcal database indicated that bovine MRSE strains were not closely related to human MRSE isolates. The occurrence of MRS, especially MRSA, in bovine mastitis in Finland was low. Most methicillin-resistant bovine CoNS are MRSE, and we found evidence of a bovine MRSE strain that may spread clonally. This is the first report of a Finnish bovine isolate of MRSAmecC ST130. The study provides a baseline for further MRS monitoring.

  17. [Antimicrobial spectrum of ceftaroline. In vitro activity against methicillin-resistant staphylococci].

    Science.gov (United States)

    Cercenado, Emilia; Morosini, María Isabel

    2014-03-01

    Because of the increase in bacterial resistance, there is a need for new antimicrobial agents. In particular, Staphylococcus aureus is a frequent cause of severe infections and has an extraordinary capacity to develop antibiotic multiresistance, including resistance to glycopeptides, linezolid, and daptomycin. Although the incidence of methicillin-resistant S. aureus (MRSA) seems to have stabilized in the last few years, its wide dissemination in healthcare settings and in the community is a cause of concern. Ceftaroline is a new broad-spectrum cephalosporin with bactericidal activity against Gram-positive bacteria, including MRSA and multidrug-resistant Streptococcus pneumoniae. In addition, this drug is active against staphylococci showing resistance to glycopeptides, linezolid, and daptomycin. The ceftaroline MIC90 against MRSA ranges from 0.5-2mg/L and that against methicillin-resistant coagulase-negative staphylococci is 0.5mg/L. Ceftaroline has also good activity against respiratory pathogens including Haemophilus influenzae and Moraxella catarrhalis. Although this drug is active against Enterobacteriaceae, it does not retain activity when these isolates produce extended-spectrum beta-lactamases, carbapenemases or hyperproduce AmpC. Ceftaroline is not active against nonfermentative Gram-negative bacilli. Ceftaroline is an interesting addition to the therapeutic armamentarium against MRSA and constitutes an important option for the treatment of polymicrobial infections caused by multidrug-resistant Gram-positive microorganisms. Copyright © 2014 Elsevier España, S.L. All rights reserved.

  18. [Nonfermentative gram-negative bacteria: isolation rates and antibiotic sensitivity].

    Science.gov (United States)

    Bogomolova, N S; Bol'shakov, L V; Kuznetsova, S M; Oreshkina, T D

    2010-01-01

    The isolation rates of nonfermentative gram-negative bacteria (NFGNB) are analyzed in the inpatients treated at the B. V. Petrovsky Russian Surgery Research Center in 2005-2009 and antibiotic resistance trends in nosocomial strains of NFGNB are traced in the above period. The study of the etiological structure of nosocomial infections has shown that the past 2 years (2008 and 2009) were marked by a clear tendency for the preponderance of gram-positive coccal pathogens (46.8 and 53.9%) with a considerable (1.5-2-fold) reduction in the proportion of representatives of enterobacteria (31.5 and 24.5%) and NFGB (13.4 and 11.3%), but with an increase in the proportion of fungi up to 7.1 and 8.6%, respectively. Among the NFGNBs, P. aeruginosa remains ohe of the most common pathogens for nosocomial infections although its portion in the number of all etiologically significant microorganisms was substantially reduced (from 13% in 2005 to 4.6% in 2009). It continues to remain one of the most common causative agents for infections of the urinary tract (e.g., after renal transplantation) and upper and lower respiratory tract (e.g. nosocomial pneumonia) and for those developing after surgical interventions (postoperative wound suppuration discharged along the drainages, from a T-sized tube, etc.). Among the NFGNBs, Acinetobacter spp. was the second frequently isolated pathogen, the isolation rate for which also decreased from 7.9% in 2005 to 2.6% in 2009. Polymyxin B and carbapenems (imipenem, meropenem, and doripenem) showed the highest activity against the vast majority of the test strains; however, there was an absolutely clear declining trend in the proportion of carbapenem-sensitive strains among virtually all the NFGNBs under study. According to the proportion of imipenem-, meropenem-, and doripenem-sensitive nosocomial P. aeroginosa strains (66.7, 46.6, and 44.7%, respectively), doripenem had the least activity. Acinetobacter spp. strains sensitive to these drugs showed

  19. Biofilm Matrix Composition Affects the Susceptibility of Food Associated Staphylococci to Cleaning and Disinfection Agents

    OpenAIRE

    Annette eFagerlund; Solveig eLangsrud; Even eHeir; Maria Ingeborg Mikkelsen; Trond eMøretrø

    2016-01-01

    Staphylococci are frequently isolated from food processing environments, and it has been speculated whether survival after cleaning and disinfection with benzalkonium chloride-containing disinfectants is due to biofilm formation, matrix composition or benzalkonium chloride efflux mechanisms. Out of 35 food associated staphylococci, eight produced biofilm in a microtiter plate assay and were identified as Staphylococcus capitis (2), S. cohnii, S. epidermidis, S. lentus (2), and S. saprophyticu...

  20. Isolation and Identification of Aroma Producing Staphylococci from Sichuan Sausage%四川香肠中产香葡萄球菌的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    杨勇; 张雪梅; 程艳; 刘书亮; 李诚; 秦文; 蒲彪

    2011-01-01

    从四川自然发酵香肠中共分离得到242株微球菌和葡萄球菌,采用快速半定量平板方法从中筛选出了硝酸盐还原酶活性较大的株菌80株,并对这些菌株进行发酵基本实验和产香基本要求实验,筛选得到了2株过氧化氢酶阳性葡萄球菌S15和S25。通过GC-MS分析发现,在实验条件下只有S25具有代谢亮氨酸生成3-甲基丁醛的能力。生化鉴定以及菌株的16S rDNA测序结果表明,S25为腐生葡萄球菌。%A total of 242 strains of Micrococcaceae and Staphylococcus were isolated from Sichuan natural fermented sausages.80 strains which have high nitrate reduction properties were selected by a fast semi-quantitative plate assay method.Different experiments were carried out according to the common selection criteria of starter culture and aroma production.Two strains of Staphylococcus named S15 and S25 were obtained with the ability to produce catalase.GC-MS analysis showed that only S25 produced 3-methyl-butanal from leucine under the experimental conditions.S25 was identified as Staphylococcus saprophyticus by its biochemical characteristics and 16S rDNA sequences.

  1. Bacteremias por Staphylococcus coagulase negativos oxacilina resistentes em um hospital escola na cidade de Santa Maria, Estado do Rio Grande do Sul Oxacillin-resistant coagulase-negative Staphylococci bacteremia at a teaching hospital in Santa Maria, State of Rio Grande do Sul, Brazil

    Directory of Open Access Journals (Sweden)

    Fabiane Rigatti

    2010-12-01

    Full Text Available INTRODUÇÃO: Neste estudo, objetivou-se caracterizar a prevalência e o perfil de suscetibilidade de cepas de Staphylococcus coagulase negatives resistentes à oxacilina isoladas de culturas de sangue, em um hospital escola, localizado na Cidade de Santa Maria. Além disso, buscou-se comparar ao teste genotípico de referência, diferentes metodologias fenotípicas para a caracterização da resistência mediada pelo gene mecA. MÉTODOS: Após identificação (MicroScan® - Siemens, os isolados foram submetidos a testes de sensibilidade antimicrobiana a partir da difusão do disco e automação (MicroScan® - Siemens. A presença do gene mecA foi evidenciada através da técnica molecular de reação em cadeia da polimerase. RESULTADOS: A espécie prevalente foi Staphylococcus epidermidis (67%. O gene mecA foi detectado em 90% das cepas e conforme análise dos perfis de sensibilidade, observou-se um índice elevado de resistência a várias classes de antimicrobianos. Contudo, todos os isolados mostraram-se uniformemente sensíveis à vancomicina e tigeciclina. O disco de cefoxitina foi a metodologia fenotípica que melhor correlacionou-se com o padrão ouro. CONCLUSÕES: A análise da significância clínica de SCN isolados de hemoculturas e a detecção precisa da resistência à oxacilina representam fatores decisivos para a instituição correta da antibioticoterapia. Apesar da vancomicina constituir o tratamento usual na maioria dos hospitais brasileiros, tem a redução de seu emprego recomendada.INTRODUCTION: This study aimed to characterize the prevalence and susceptibility profile to oxacillin-resistant Coagulase-negative Staphylococci strains isolated from blood cultures in a teaching hospital, located in Santa Maria, RS. In addition, different methodologies for phenotypic characterization of mecA-mediated oxacillin resistance were compared with genotypic reference testing. METHODS: After identification (MicroScan® - Siemens, the

  2. Occurrence of cfr-mediated multiresistance in staphylococci from veal calves and pigs, from humans at the corresponding farms, and from veterinarians and their family members.

    Science.gov (United States)

    Cuny, Christiane; Arnold, Phillippe; Hermes, Julia; Eckmanns, Tim; Mehraj, Jaishri; Schoenfelder, Sonja; Ziebuhr, Wilma; Zhao, Qin; Wang, Yang; Feßler, Andrea T; Krause, Gérard; Schwarz, Stefan; Witte, Wolfgang

    2017-02-01

    This study reports on the emergence of linezolid-resistant coagulase-negative staphylococci (CoNS) containing the multiresistance gene cfr in veal calves and pigs, as well as in humans exposed to these animals. CoNS (Staphylococcus auricularis, Staphylococcus cohnii, Staphylococcus lentus, Staphylococcus kloosii, Staphylococcus sciuri, Staphylococcus simulans), but not Staphylococcus aureus, carrying the gene cfr were detected in samples of 12 out of 52 calves at three farms which had a history of florfenicol use. Nasal swabs from 10 humans living on these farms were negative for cfr-carrying staphylococci. Nasal swabs taken from 142 calves at 16 farms in the same area that did not use florfenicol were also negative for cfr-carrying staphylococci. 14 cfr-carrying CoNS (S. kloosii, S. saprophyticus, S. simulans) were detected in three of eight conventional pig farms investigated. One of 12 humans living on these farms harboured a cfr-carrying S. cohnii. Among the nasal swabs taken from 169 veterinarians from all over Germany, four (2.3%) were positive for cfr-carrying CoNS (three S. epidermidis, one S. saprophyticus), and three (1.1%) of 263 contact persons of this group also harboured cfr-carrying CoNS (one S. epidermidis, two S. saprophyticus). In vitro conjugation of cfr by filter mating to S. aureus 8325-4 was possible for 10 of 34CoNS and the cfr gene was associated with plasmids of 38-40kb. Moreover, a total of 363 humans of a German municipal community were investigated for nasal carriage of cfr-carrying staphylococci to get an idea whether such isolates are disseminated as nasal colonizers in non-hospitalized humans in the community, were all negative. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Challenges in the Prevention of Coagulase-Negative Staphylococcal Sepsis in Neonates

    NARCIS (Netherlands)

    V. Hira (Vishal)

    2013-01-01

    textabstractCoagulase-negative staphylococci (CoNS) are the leading cause of late-onset sepsis (LOS) worldwide. As antibiotic resistance is dramatically increasing in these organisms, prevention is becoming more and more important. CoNS that are isolated from blood cultures of neonates with LOS on n

  4. Coagulase-negative staphylococcal skin carriage among neonatal intensive care unit personnel: From population to infection

    NARCIS (Netherlands)

    V. Hira (Vishal); M. Sluijter (Marcel); W.H.F. Goessens (Wil); A. Ott (Alewijn); R. de Groot (Ronald); P.W.M. Hermans (Peter); R.F. Kornelisse (René)

    2010-01-01

    textabstractCoagulase-negative staphylococci (CoNS) are a major cause of sepsis in neonatal intensive care units (NICU) worldwide. Infecting strains of these commensal bacteria may originate from NICU personnel. Therefore, we studied the characteristics of CoNS isolates from NICU personnel and compa

  5. Coagulase-negative staphylococcal skin carriage among neonatal intensive care unit personnel: from population to infection.

    NARCIS (Netherlands)

    Hira, V.; Sluijter, M.; Goessens, W.H.F.; Ott, A.; Groot, R. de; Hermans, P.W.M.; Kornelisse, R.F.

    2010-01-01

    Coagulase-negative staphylococci (CoNS) are a major cause of sepsis in neonatal intensive care units (NICU) worldwide. Infecting strains of these commensal bacteria may originate from NICU personnel. Therefore, we studied the characteristics of CoNS isolates from NICU personnel and compared them to

  6. Detection of Staphylococcus aureus among coagulase positive staphylococci from animal origin based on conventional and molecular methods

    Directory of Open Access Journals (Sweden)

    Nikolina Velizarova Rusenova

    2017-03-01

    Full Text Available The present study aimed to detect Staphylococcus aureus (S. aureus among other coagulase positive staphylococci from animal origin by using conventional methods (biochemical tests and latex agglutination and a molecular method, based on the nuc gene, as the gold standard and to assess the usefulness of these methods. For this purpose, total of 344 staphylococcal isolates were collected and analysed. A total of 156 isolates suspicious for S. aureus were detected by a conventional biochemical method – 88 from cows, 18 from goats, 7 from pigs, 17 from poultry, 7 from rabbits and 19 from dogs. The majority of S. aureus strains gave typical biochemical reactions with the exception of 30 (19.2% and 25 (16% that were VP negative and weak positive in fermenting mannitol, respectively. Twelve strains were found to be non-haemolytic (7.7% and four strains did not ferment trehalose (2.6%. Other staphylococci were identified as S. pseudintermedius (n = 103, S. hyicus (n = 23 and the rest were coagulase-negative staphylococci. Latex agglutination test resulted in rapid positive reactions with S. aureus with exception of 5 strains (3.2% from cow mastitis milk. Positive agglutination reactions were also established with S. pseudintermedius, and S. hyicus. PCR confirmed all strains that were preliminary identified as S. aureus by amplification of 270 bp fragment of nuc gene specific for this species. The atypical reactions in certain strains established in this study have shown that the precise detection of S. aureus from animal origin should be done by combination of conventional and molecular methods.

  7. Characterization of staphylococci and their role in pathology of children

    Directory of Open Access Journals (Sweden)

    M. A. Harуfulina

    2014-03-01

    Full Text Available The problem of human staphylococcal infection in the world has been considered. It is known that there are 27 species of staphylococci, 14 of them colonize the skin and mucous membranes of humans, and 3 of them may cause diseases. They are Staphylococcus aureus, S. epidermidis and S. saprophyticus. Staphylococci are causative agents of the large part of community-acquired and nosocomial bacteremia, pneumonia, infections of skin and soft tissues, bones and joints. Of all cases of nosocomial infections, S. aureus accounts for 31%. During the identification of 65 strains of bacteria isolated from people with respiratory tract lesions, it was found that 58.5% of cases were connected with presence of staphylococci. 86.8% of isolates identified belong to species S. aureus and 13.2% to S. saprophyticus. Study of pathogenicity factors manifestation revealed that all strains identified as S. aureus had plasmocoagulase; lipase and lecitinase were simultaneously detected in 78.3% of them, among S. saprophyticus strains in 20%. Presence of haemolysins was determined for 82.1% of strains of staphylococci. Ability to film formation was detected in 21 (63.6% strains of S. aureus and 2 (40% of S. saprophyticus strains. Given high frequency of carriers of staphylococci among healthy individuals, they should be considered as potentially pathogenic microorganisms and opportunistic pathogens. Due to their pathogenicity factors (adhesins, capsule, cell wall components, enzymes and toxins, staphylococci may cause diseases such as sepsis, meningitis, peritonitis, endocarditis, pneumonia, tonsillitis, and in young children they may result in serious Ritter illness (“babies scalded syndrome”, foodstuff infection and staphylococcal enterocolitis, in lactating women – mastitis, toxic shock syndrome. To solve the problem of staphylococcal infection prevention, it is necessary to use the following prophylaxis methods: to avoid decrease in immunity, to observe the

  8. A molecular epidemiological study of methicillin-resistant Staphylococci environmental contamination in railway stations and coach stations in Guangzhou of China.

    Science.gov (United States)

    Lin, J L; Peng, Y; Ou, Q T; Lin, D X; Li, Y; Ye, X H; Zhou, J L; Yao, Z J

    2017-02-01

    Methicillin-resistant Staphylococcus aureus (MRSA) has caused a series of public health problems since it was first found in 1961. However, there are few research studies on the MRSA environmental contamination in railway stations and coach stations. Therefore, the aim of this study was to determine MRSA environmental contamination in public transport stations. Between December 2013 and January 2014, 380 surface samples from three railway stations (180) and four coach stations (200) in Guangzhou were collected to isolate and determine the prevalence and characteristics of Staphylococci strains. 39·21% of all samples were Staphylococci isolates, 1·58% of Staphylococci isolates were MRSA isolates, and 6·05% were methicillin-susceptible S. aureus. The proportion of multidrug resistant among 149 Staphylococci isolates was 75·84%. None of MRSA isolates was identified with the Panton-Valentine Leukocidin (PVL) genes, and one of them was identified with the qac gene. Four MRSA isolates were Staphylococcal Cassette Chromosome mec IVa, and the other two were nontypeable. Staphylococcus aureus isolates were classified into several sequence types (STs), and STs showed possible cross-transmissions of isolates from various sources. Methicillin-resistant Staphylococci contamination prevalence was high, and the environment of stations may be the vectors transmitting the Staphylococci to passengers. This is the first study to comprehensively report the prevalence, antibiotic resistance, and molecular characteristics of contamination of Staphylococci isolates in railway stations and coach stations of China. It will have great public health implications on infection control in community settings because of the serious hazard of Staphylococci, especially methicillin-resistant Staphylococci. Our findings have provided evidence for relevant departments to reduce the contamination of Staphylococci in environment of public transport stations. © 2016 The Society for Applied

  9. Isolated port site recurrence of node-negative clinical stage IB1 cervical adenocarcinoma

    Directory of Open Access Journals (Sweden)

    Uma Deshmukh

    2017-05-01

    Conclusion: This is the first case report describing an isolated port site recurrence in a patient who underwent robotic-assisted laparoscopic surgery for early-stage cervical adenocarcinoma with negative margins and negative lymph nodes. The mechanism underlying this isolated recurrence remains unknown.

  10. Biodegradative activities of some gram- negative bacilli isolated ...

    African Journals Online (AJOL)

    The biodegradative activities of three Gram negative bacilli (Aeromonas ... Cowpea seeds of the variety Samaru-40 were cultivated and observed for eight weeks. ... of germination, plant height, number of leaves, leaf area and root nodules.

  11. Detection of -Lactamases in nosocomial gram negative clinical isolates

    Directory of Open Access Journals (Sweden)

    Rodrigues C

    2004-01-01

    Full Text Available -lactamases represent the most common mechanism of -lactam resistance. Extended spectrum -lactamases (ESBLs represent a major group of -lactamses currently being identified worldwide in large numbers along with inducible AmpC -lactamases and derepressed mutants. The present study was done to detect -lactamase production in clinical isolates by rearranging routine discs used in reporting susceptibility to specifically assess ESBLs, AmpC -lactamases (both inducible and hyperproducers i.e., derepressed mutants. A total of 286 clinical isolates were studied using a novel predictor disc approximation method to detect the above mechanisms of resistance with careful use and placement of antimicrobial discs. Of the 286 isolates, 151(53% were ESBL producers of which 131(46% were also derepressed mutants while remaining 20(7% were plain ESBL producers. Forty (14% were plain derepressed mutants. Inducible AmpC -lactamase production was detected in 19(7% of the isolates. The commonest ESBL producers were E.coli and K. pneumoniae. The high incidence of -lactamase production due to multiple mechanisms in clinical isolates is alarming and urgent action needs to be taken from both a therapeutic and infection control perspective.

  12. Draft Genome Sequence of Staphylococcus simulans UMC-CNS-990, Isolated from a Case of Chronic Bovine Mastitis.

    Science.gov (United States)

    Calcutt, Michael J; Foecking, Mark F; Hsieh, Hsin-Yeh; Perry, Jeanette; Stewart, George C; Middleton, John R

    2013-12-12

    Coagulase-negative staphylococci are frequently isolated from cases of subclinical bovine mastitis. Reported here is a draft genome sequence of Staphylococcus simulans UMC-CNS-990, an isolate recovered from a chronic intramammary infection of a Holstein cow. Unexpectedly, a cluster of genes encoding gas vesicle proteins was found within the 2,755-kb genome.

  13. Phenotypic and genotypic characterization of prolyliminopeptidase-negative Neisseria gonorrhoeae isolates in Denmark

    DEFF Research Database (Denmark)

    Fjeldsøe-Nielsen, H; Unemo, M; Fredlund, H;

    2005-01-01

    In the study presented here 26 recent Danish clinical isolates of prolyliminopeptidase (PIP)-negative Neisseria gonorrhoeae were phenotypically and genotypically characterized to investigate whether one or more PIP-negative strains are circulating in the Danish community. The profiles...... of these isolates were compared with those of three isolates from a recent outbreak of PIP-negative N. gonorrhoeae infection in the UK. Twenty-five of the Danish isolates and all three UK isolates had similar antibiograms and were designated serovar IB-4. Genotypic characterization by pulsed-field gel...... electrophoresis, porB1b gene sequencing, and opa-typing revealed that these isolates were indistinguishable or closely related. The results indicate that at least one PIP-negative N. gonorrhoeae strain is currently circulating in the Danish community, and this strain is indistinguishable from the one that caused...

  14. Methicillin-resistant staphylococci: implications for our food supply?

    Science.gov (United States)

    Doyle, M Ellin; Hartmann, Faye A; Lee Wong, Amy C

    2012-12-01

    Food-borne intoxication, caused by heat-stable enterotoxins produced by Staphylococcus aureus, causes over 240,000 cases of food-borne illness in the United States annually. Other staphylococci commonly associated with animals may also produce these enterotoxins. Foods may be contaminated by infected food handlers during slaughter and processing of livestock or by cross-contamination during food preparation. S. aureus also causes a variety of mild to severe skin and soft tissue infections in humans and other animals. Antibiotic resistance is common in staphylococci. Hospital-associated (HA) S. aureus are resistant to numerous antibiotics, with methicillin-resistant S. aureus (MRSA) presenting significant challenges in health care facilities for over 40 years. During the mid-1990s new human MRSA strains developed outside of hospitals and were termed community-associated (CA). A few years later, MRSA was isolated from horses and methicillin resistance was detected in Staphylococcus intermedius/pseudintermedius from dogs and cats. In 2003, a livestock-associated (LA) MRSA strain was first detected in swine. These methicillin-resistant staphylococci pose additional food safety and occupational health concerns. MRSA has been detected in a small percentage of retail meat and raw milk samples indicating a potential risk for food-borne transmission of MRSA. Persons working with animals or handling meat products may be at increased risk for antibiotic-resistant infections. This review discusses the scope of the problem of methicillin-resistant staphylococci and some strategies for control of these bacteria and prevention of illness.

  15. Genetic regulation of the intercellular adhesion locus in staphylococci

    Directory of Open Access Journals (Sweden)

    David R Cue

    2012-03-01

    Full Text Available The formation of biofilms by Staphylococcus aureus and Staphylococcus epidermidis is an important aspect of many staphylococcal infections, most notably endocarditis, osteomyelitis and infections associated with indwelling medical devices. The major constituents of S. aureus biofilms are polysaccharides, such as poly N-acetyl glucosamine (PIA/PNAG, cell surface and secreted bacterial proteins, and extracellular DNA. The exact composition of biofilms often varies considerably between different strains of staphylococci and between different sites of infection by the same strain. PIA/PNAG is synthesized by the products of 4 genes, icaADBC, that are encoded in a single operon. A fifth gene, icaR, is a negative regulator of icaADBC. Expression of icaADBC is tightly regulated, but can often be induced in vitro by growing staphylococci in the presence of high salt, high glucose or ethanol. Regulation of icaADBC is complex and numerous regulatory factors have been implicated in control of icaADBC. Many of these are well known global transcriptional regulatory factors like SarA and sigmaB, whereas other regulators, such as IcaR, seem to affect expression of relatively few genes. Here, we will attempt to summarize how various regulatory factors affect the production of PIA/PNAG in staphylococci.

  16. Prevalence of Genotypes That Determine Resistance of Staphylococci to Macrolides and Lincosamides in Serbia

    Directory of Open Access Journals (Sweden)

    Milena Mišić

    2017-08-01

    Full Text Available Macrolides, lincosamides, and streptogramins (MLS resistance genes are responsible for resistance to these antibiotics in Staphylococcus infections. The purpose of the study was to analyze the distribution of the MLS resistance genes in community- and hospital-acquired Staphylococcus isolates. The MLS resistance phenotypes [constitutive resistance to macrolide–lincosamide–streptogramin B (cMLSb, inducible resistance to macrolide–lincosamide–streptogramin B (iMLSb, resistance to macrolide/macrolide–streptogramin B (M/MSb, and resistance to lincosamide–streptogramin A/streptogramin B (LSa/b] were determined by double-disc diffusion method. The presence of the MLS resistance genes (ermA, ermB, ermC, msrA/B, lnuA, lnuB, and lsaA were determined by end-point polymerase chain reaction in 179 isolates of staphylococci collected during 1-year period at the Center for Microbiology of Public Health Institute in Vranje. The most frequent MLS phenotype among staphylococcal isolates, both community-acquired and hospital-acquired, was iMLSb (33.4%. The second most frequent was M/MSb (17.6% with statistically significantly higher number of hospital-acquired staphylococcal isolates (p < 0.05. MLS resistance was mostly determined by the presence of msrA/B (35.0% and ermC (20.8% genes. Examined phenotypes were mostly determined by the presence of one gene, especially by msrA/B (26.3% and ermC (14.5%, but 15.6% was determined by a combination of two or more genes. M/MSb phenotype was the most frequently encoded by msrA/B (95.6% gene, LSa/b phenotype by lnuA (56.3% gene, and iMLSb phenotype by ermC (29.4% and ermA (25.5% genes. Although cMLSb phenotype was mostly determined by the presence of ermC (28.9%, combinations of two or more genes have been present too. This pattern was particularly recorded in methicillin-resistant Staphylococcus aureus (MRSA (58.3% and methicillin-resistant coagulase-negative staphylococci (MRCNS (90.9% isolates with c

  17. Antibiotic Exposure in a Low-Income Country: Screening Urine Samples for Presence of Antibiotics and Antibiotic Resistance in Coagulase Negative Staphylococcal Contaminants

    DEFF Research Database (Denmark)

    Lerbeck, Anne Mette; Tersbøl, Britt Pinkowski; Styrishave, Bjarne

    2014-01-01

    Development of antimicrobial resistance has been assigned to excess and misuse of antimicrobial agents. Staphylococci are part of the normal flora but are also potential pathogens that have become essentially resistant to many known antibiotics. Resistances in coagulase negative staphylococci (Co...... was the most common isolate (75%), followed by S. epidermidis (13%) and S. hominis (6%). S. haemolyticus was also the species displaying the highest resistance prevalence (82%). 69% of the isolated CoNS were multiple drug resistant (§4 antibiotics) and 45% of the CoNS were methicillin resistant. Antimicrobial...

  18. Antibiotic Exposure in a Low-Income Country: Screening Urine Samples for Presence of Antibiotics and Antibiotic Resistance in Coagulase Negative Staphylococcal Contaminants

    DEFF Research Database (Denmark)

    2014-01-01

    Development of antimicrobial resistance has been assigned to excess and misuse of antimicrobial agents. Staphylococci are part of the normal flora but are also potential pathogens that have become essentially resistant to many known antibiotics. Resistances in coagulase negative staphylococci (Co...... was the most common isolate (75%), followed by S. epidermidis (13%) and S. hominis (6%). S. haemolyticus was also the species displaying the highest resistance prevalence (82%). 69% of the isolated CoNS were multiple drug resistant (≧4 antibiotics) and 45% of the CoNS were methicillin resistant. Antimicrobial...

  19. Virulence of pertactin-negative Bordetella pertussis isolates from infants, France.

    Science.gov (United States)

    Bodilis, Hélène; Guiso, Nicole

    2013-03-01

    Bordetella pertussis isolates that do not express pertactin (PRN) are increasing in regions where acellular pertussis vaccines have been used for >7 years. We analyzed data from France and compared clinical symptoms among infants <6 months old infected by PRN-positive or PRN-negative isolates. No major clinical differences were found between the 2 groups.

  20. Isolated Ileal Stricture Secondary to Antigen-Negative GI Histoplasmosis in a Patient on Immunosuppressive Therapy

    Science.gov (United States)

    Green, Michael; Nehme, Fredy; Tofteland, Nathan

    2017-01-01

    We present a case of antigen-negative disseminated histoplasmosis manifesting as an isolated ileal stricture in a patient on chronic infliximab and methotrexate. Diagnosis can be challenging due to imperfect tests, and this condition should remain in the differential, even with negative testing. Mortality of untreated disseminated histoplasmosis can be as high as 80%. PMID:28144615

  1. Topical Bactroban (mupirocin): efficacy in treating burn wounds infected with methicillin-resistant staphylococci.

    Science.gov (United States)

    Strock, L L; Lee, M M; Rutan, R L; Desai, M H; Robson, M C; Herndon, D N; Heggers, J P

    1990-01-01

    Bacterial antimicrobial susceptibility predictors such as the minimal inhibitory concentration (MIC) assay and Nathans Agar Well Diffusion (NAWD) assay provide essential information relevant to the therapeutic approach in burn-wound sepsis. The susceptibilities of 68 gram-positive burn-wound isolates were tested against topical Bactroban (mupirocin) (Beecham Laboratories, Bristol, Tenn.) and compared with other topical antimicrobials such as mafenide acetate, silver sulfadiazine, and bacitracin/neomycin/polymyxin (BNP). Topical susceptibility data were obtained with a modification of NAWD assay. Bactroban's antimicrobial activity was greater than that of mafenide acetate (100% vs 97%), and significantly greater than that of silver sulfadiazine and that of BNP (p less than 0.001). Of the 68 isolates that were susceptible to Bactroban, 51 were predominately methicillin-resistant staphylococci (MRSA). Bactroban showed in vitro activity against 71% of the 85 gram-negative isolates tested. Mafenide acetate showed activity against 89% of these isolates, a significant difference compared with Bactroban (p less than 0.02). In general, no significant difference was found between the activities of Bactroban and silver sulfadiazine against the gram-negative isolates. The activities of mafenide acetate and silver sulfadiazine against isolates of Pseudomonas aeruginosa were significantly greater than that of Bactroban (p less than 0.05). Bactroban may be used in the treatment of documented staphylococcal burn-wound infections. On the basis of the in vitro data, 13 patients with MRSA burn-wound infections susceptible to Bactroban were evaluated. Quantitative wound biopsies were employed to determine the efficacy of this therapeutic approach. The outcome of these infections was correctly predicted by the NAWD assay in 92.3% of the patients treated (p less than 0.0005).(ABSTRACT TRUNCATED AT 250 WORDS)

  2. Fuzzy sliding mode controller of a pneumatic active isolating system using negative stiffness structure

    Energy Technology Data Exchange (ETDEWEB)

    Le, Thanh Danh; Ahn, Kyoung Kwan [Univ. of Ulsan, Ulsan (Korea, Republic of)

    2012-12-15

    A novel active vibration isolation system using negative stiffness structure (active system with NSS) for low excitation frequency ranges (< 5 Hz) is developed successfully. Here, the negative stiffness structure (NSS) is used to minimize the attraction of vibration. Then, the fuzzy sliding mode controller (FSMC) is designed to improve the vibration isolation performance of the active system with NSS. Based on Lyapunov stability theorem, the fuzzy control rules are constructed. Next, the experimental apparatus is built for evaluating the isolation efficiency of the proposed system controlled by the FSMC corresponding to various excitation conditions. In addition, the isolation performance of the active system with NSS, the active system without NSS and the passive the system with NSS is compared. The experimental results confirmed that the active system with NSS gives better isolation efficiency than the active system without NSS and the passive system with NSS in low excitation frequency areas.

  3. Prevalence and risk factors for carriage of multi-drug resistant Staphylococci in healthy cats and dogs.

    Science.gov (United States)

    Gandolfi-Decristophoris, Paola; Regula, Gertraud; Petrini, Orlando; Zinsstag, Jakob; Schelling, Esther

    2013-01-01

    We investigated the distribution of commensal staphylococcal species and determined the prevalence of multi-drug resistance in healthy cats and dogs. Risk factors associated with the carriage of multi-drug resistant strains were explored. Isolates from 256 dogs and 277 cats were identified at the species level using matrix-assisted laser desorption ionisation-time of flight mass spectrometry. The diversity of coagulase-negative Staphylococci (CNS) was high, with 22 species in dogs and 24 in cats. Multi-drug resistance was frequent (17%) and not always associated with the presence of the mecA gene. A stay in a veterinary clinic in the last year was associated with an increased risk of colonisation by multi-drug resistant Staphylococci (OR = 2.4, 95% CI: 1.1~5.2, p value LRT = 0.04). When identifying efficient control strategies against antibiotic resistance, the presence of mechanisms other than methicillin resistance and the possible role of CNS in the spread of resistance determinants should be considered.

  4. [In vitro activity of roxithromycin compared to 5 other macrolides against staphylococci].

    Science.gov (United States)

    Brun, Y; Coulet, M; Forey, F; Fleurette, J

    1988-06-01

    This study aimed to compare the bacteriostatic activity of roxithromycin (RU) to those of erythromycin (ERY), troleandomycin (TAO), spiramycin (SPI), josamycin (JOS) and midekamycin (MID) against staphylococci strains. 239 strains of hospital origin were analysed: S. aureus (139), coagulase negative staphylococci (CNS) (100). The MIC were determined by the agar dilution method. The modal MIC, the MIC 50 and 90 observed for the both groups of strains are given according to species and antibiotics. This study gives the opportunity to classify the 6 antibiotics in a decreasing order considering their antistaphylococcal activity: RU = ERY, TAO = SPI, JOS = MID. No difference was noticeable between S. aureus and CNS strains.

  5. Correlation between clinical data and antibiotic resistance in coagulase-negative Staphylococcus species isolated from 68 patients with acute post-cataract endophthalmitis.

    Science.gov (United States)

    Chiquet, C; Maurin, M; Altayrac, J; Aptel, F; Boisset, S; Vandenesch, F; Cornut, P L; Romanet, J P; Gain, P; Carricajo, A

    2015-06-01

    Coagulase-negative staphylococci (CNS) cause the majority of post-cataract endophthalmitis, which can lead to anatomical and/or functional loss of the eye. This study reports the antibiotic susceptibilities of CNS isolates associated with acute post-cataract endophthalmitis cases and correlates antibiotic resistance with severity and outcome of infection in these patients. Clinical data (initial ocular examination, final prognosis, antibiotic treatment) and the antibiotic susceptibilities of the isolated CNS strains were obtained from 68 patients with post-surgical endophthalmitis recruited during a 7-year period by the FRench Institutional ENDophthalmitis Study (FRIENDS) group. The CNS strains displayed 100% susceptibility to vancomycin, 70% to fluoroquinolones, 83% to fosfomycin, 46% to imipenem and 18% to piperacillin. The most effective antibiotic combinations were fosfomycin plus a fluoroquinolone and imipenem plus a fluoroquinolone, which were considered adequate in 80% and 58% of patients, respectively. Methicillin resistance was significantly associated with older age (p 0.001), diabetes mellitus (p 0.004), absence of fundus visibility (p 0.06), and poor visual prognosis (p 0.03). Resistance to fluoroquinolones was significantly associated with absence of fundus visibility (p 0.05) and diabetes mellitus (p 0.02). This large prospective study demonstrates that methicillin resistance and, to a lesser extent, fluoroquinolone resistance in CNS strains causing postoperative endophthalmitis are both prevalent in France and associated with a poorer visual prognosis. These results emphasize the need for an effective surveillance of this antibiotic resistance and the development of new diagnostic tools for rapid detection for early optimization of antibiotic therapy in endophthalmitis patients.

  6. Antibiotic Resistance of Gram Negative Bacteria Isolated From Urine Cultures in Our Laboratory

    Directory of Open Access Journals (Sweden)

    Hakan Temiz

    2008-12-01

    Full Text Available In this study; we analyzed the antimicrobial susceptibility of Gram negative bacteria isolated from urine cultures in the Microbiology Laboratory of Dicle University Medical Faculty Hospital from January 2006 to December 2006; retrospectively. Escherichia coli and Klebsiella species were the most frequently isolated bacteria from both outpatients and hospitalized patients. The most effective antibiotics to these bacteria were carbapenems. These results were suggested to be useful for empirical treatment of urinary system infections in our hospital.

  7. Methicillin-resistant Staphylococci in companion animals

    DEFF Research Database (Denmark)

    Baptiste, Keith Edward; Williams, Kerry; Williams, Nicola J.

    2005-01-01

    We determined the molecular characteristics of methicillin-resistant staphylococci from animals and staff at a small animal and equine hospital. Methicillin-restistant Staphylococcus aureus identical to human EMRSA-15 was found in dogs and hospitalstaff. In contrast, 5 distinct MRSA strains were...

  8. 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.

  9. Prevalence and genetic analysis of phenotypically Vi- negative Salmonella typhi isolates in children from Kathmandu, Nepal.

    Science.gov (United States)

    Pulickal, Anoop S; Callaghan, Martin J; Kelly, Dominic F; Maskey, Mitu; Mahat, Sandeep; Hamaluba, Mainga; Dongol, Sabina; Adhikari, Neelam; Thorson, Stephen; Basynat, Buddha; Murdoch, David R; Farrar, Jeremy J; Pollard, Andrew J

    2013-08-01

    The Vi capsular polysaccharide (ViPS) protects Salmonella enterica subspecies enterica serotype Typhi (S.Typhi) in vivo by multiple mechanisms. Recent microbiological reports from typhoid endemic countries suggest that acapsulate S.Typhi may occur in nature and contribute to clinical typhoid fever that is indistinguishable from disease caused by capsulate strains. The prevalence and genetic basis of ViPS-negative S.Typhi isolates in children from Kathmandu, Nepal, were tested in 68 isolates. Although 5.9% of isolates tested negative for capsular expression by slide agglutination tests, a novel multiplex PCR assay and individual PCR analyses demonstrated the presence of all 14 genes responsible for the synthesis, transportation and regulation of the ViPS. These data suggest that phenotypically acapsulate S.Typhi may not have a genetic basis for the same.

  10. Cohabitation--relationships of corynebacteria and staphylococci on human skin.

    Science.gov (United States)

    Kwaszewska, Anna; Sobiś-Glinkowska, Maria; Szewczyk, Eligia M

    2014-11-01

    Skin microbiome main cultivable aerobes in human are coagulase-negative staphylococci and lipophilic corynebacteria. Staphylococcus strains (155) belonging to 10 species and 105 strains of Corynebacterium belonging to nine species from the skin swabs of healthy male volunteers were investigated to determine their enzymatic activity to main metabolic substrates: carbohydrates, proteins, lipids, and response to factors present on the skin such as osmotic pressure, pH, and organic acids. The results showed that lipophilic corynebacteria have different capacity for adaptation on the skin than staphylococci. Most of Corynebacterium spp. expressed lack of proteinase, phospholipase, and saccharolytic enzymes activity. Corynebacteria were also more sensitive than Staphylococcus spp. to antimicrobial agents existing on human skin, especially to low pH. These characters can explain domination of Staphylococcus genera on healthy human skin. It can be suggested that within these two bacterial genus, there exists conceivable cooperation and reciprocal protection which results in their quantitative ratio. Such behavior must be considered as crucial for the stability of the population on healthy skin.

  11. 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.

  12. 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

  13. An Ultra-Low Frequency Two DOFs’ Vibration Isolator Using Positive and Negative Stiffness in Parallel

    Directory of Open Access Journals (Sweden)

    Min Wang

    2016-01-01

    Full Text Available With the improvement of performance in the ultra-precision manufacturing engineering, the requirements for vibration isolation have become increasingly stringent. In order to get wider effective bandwidth and higher performance of vibration isolation in multiple DOFs system, an ultra-low frequency two DOFs’ vibration isolator with positive and negative stiffness in parallel (PNSP is proposed. The two DOFs’ isolator which combines a positive stiffness (PS air spring with a negative stiffness (NS magnetic spring in parallel and combines a PS flat spring with an NS inverted pendulum in parallel is designed to reduce the natural frequency and broaden the effective bandwidth in horizontal and vertical direction. Based on this structure, stiffness models of different components in different directions are established. Compared with a PS isolator, it possesses the characteristic of high-static-low-dynamic stiffness. The simulation curves also provide strong evidence. Last, a real-time active control system and a spectrum testing and analysis system are used for the contrast experiment between the mentioned PNSP structure and PS only. The experimental results demonstrate that the isolator with PNSP can obviously reduce the natural frequency to 1 Hz and simultaneously maintain the stability of the system and consequently verify the validity and superiority of the mentioned structure.

  14. Isolamento de coliformes, estafilococos e enterococos de leite cru provenientes de tanques de refrigeração por expansão comunitários: identificação, ação lipolítica e proteolítica Isolation of coliforms, staphylococci, and enterococci in raw milk from communitarian expansion refrigeration tanks: identification, lipolytic and proteolytic action

    Directory of Open Access Journals (Sweden)

    Victor Maximiliano Reis Tebaldi

    2008-09-01

    health, since, depending on the species isolated, directed actions may be taken to improve milk quality. Milk deterioration is mainly the result of the growth of psychrotrophic microorganisms that produce heat-resistant lipases and proteases which are not denatured during pasteurization, conferring rancid and sour flavors, respectively. Thus, this work aimed at isolating and identifying the main genera of bacteria belonging to the Enterobacteriaceae family, oxidase-positive Gram-negative, Staphylococcus and Enterococcus genera, as well as determining lipase and protease activity in 16 rural farms in Boa Esperança-MG, Brazil. The Gram-negative bacteria were isolated in Eosin Methylene Blue (EMB and Hektoen Enteric agar media. Staphylococci were isolated in Baird-Parker agar and Enterococcus in KF agar. Colonies of interest were collected and submitted to Gram stain and to catalase and oxidase tests. Following these procedures, the isolates selected were identified using Bactray I, II and III; Api 20 Strep and tests suggested by Bergey's Manual of Determinative Bacteriology. Sorological identification of Enterococcus was carried out using Prolex. Milk from the 16 rural properties contained strains of fecal microorganisms, such as Escherichia coli and Enterococcus from the Lancefield D group. Oxidase-positive Gram-negative bacteria were identified in five farms. Staphylococcus was found in 10 farms. Milk collected from the farms investigated contained microorganisms that compromise its quality. All the microorganism groups tested showed lipase and protease activities.

  15. Molecular characterization and antimicrobial susceptibility of tcdA-negative Clostridium difficile isolates from Guangzhou, China.

    Science.gov (United States)

    Lidan, Chen; Linhai, Li; Yang, Liao; Zhaohui, Sun; Xiaoyan, Huang; Yuling, Shi

    2016-04-01

    This study aimed to investigate the molecular characteristics and antimicrobial susceptibility of Clostridium difficile clinical isolates in Guangzhou, China. One hundred twenty isolates were collected from Guangzhou General Hospital at the Guangzhou Military Command in China from March 2014 to April 2015, and 9 isolates were identified as tcdA-negative/tcdB-positive (A(-)B(+)) strains. Results showed that all of the strains were confirmed to be ST37 and 0 single nucleotide variants (SNVs) were found in the PaLoc region, and >60 SNVs were identified throughout the whole genome sequence. The results show the diversity of the antibiotic and gene mutations present in these strains. All of the A(-)B(+) isolates were highly resistant to clindamycin and erythromycin; showed an average sensitivity to fluoroquinolones; and maintained a high susceptibility to metronidazole, vancomycin, and tigecycline.

  16. Effects of intra-mammary bacterial infection with coagulase negative staphylococci and stage of lactation on shedding of epithelial cells and infiltration of leukocytes into milk: comparison among cows, goats and sheep.

    Science.gov (United States)

    Leitner, Gabriel; Merin, Uzi; Krifucks, Oleg; Blum, Shlomo; Rivas, Ariel L; Silanikove, Nissim

    2012-06-30

    The effects of mammary gland bacterial infection and stage of lactation on leukocyte infiltration into the mammary gland were compared among cows, goats and sheep. Animals were at two stages of lactation: mid or late. In mid-lactation animals, bacterial-free glands and coagulase negative Staphylococcus (CNS)-infected glands were compared. In late lactation only uninfected glands were studied. Of mid-lactation bacteria-free animals, goats had the highest number of leukocytes and % polymorphonuclears (PMNs), whereas sheep had the lowest and leukocytes number in cows were intermediate between sheep and goats. Based on %PMN, two cell clusters were found in sheep, which overlapped with the parallel cell clusters of cows and goats, but with a slightly higher number of leukocytes in each cell cluster. At late lactation, goats had higher values for %PMN and leukocyte numbers in comparison to cows, which had a similar cellular profile to sheep. The cellular immune response to CNS infection was similar for the three animal species, although the number of cells was different, while the basal cell level at mid-lactation and especially at the end of lactation was species specific.

  17. Characterization of sucrose-negative Pasteurella multocida variants, including isolates from large-cat bite wounds

    DEFF Research Database (Denmark)

    Christensen, Henrik Grimmig; Bisgaard, Magne; Angen, Øystein;

    2005-01-01

    To validate the identification of Pasteurella multocida-like bacteria negative for acid formation from sucrose, including isolates from bite wounds caused by large cats, 17 strains were phenotypically and genotypically characterized. Phylogenetic analysis of partially sequenced rpoB and infB gene...... care should be taken in the identification of Pasteurella-like bacteria isolated from bite wounds caused by large cats. The evidence of phenotypic and genotypic divergence calls for the further development of PCR tests and DNA sequencing to document doubtful isolates.......To validate the identification of Pasteurella multocida-like bacteria negative for acid formation from sucrose, including isolates from bite wounds caused by large cats, 17 strains were phenotypically and genotypically characterized. Phylogenetic analysis of partially sequenced rpoB and infB genes...... and the reference strains of P. multocida. Two strains isolated from leopard bite wounds were related to the type strain of P. dagmatis; however, they represented a new taxon (taxon 46 of Bisgaard), in accordance with their distinct phenotypic and genotypic identifications. The present study documents that sucrose...

  18. Antimicrobial Resistance and Molecular Investigation of H2S-Negative Salmonella enterica subsp. enterica serovar Choleraesuis Isolates in China.

    Science.gov (United States)

    Xie, Jing; Yi, Shengjie; Zhu, Jiangong; Li, Peng; Liang, Beibei; Li, Hao; Yang, Xiaoxia; Wang, Ligui; Hao, Rongzhang; Jia, Leili; Wu, Zhihao; Qiu, Shaofu; Song, Hongbin

    2015-01-01

    Salmonella enterica subsp. enterica serovar Choleraesuis is a highly invasive pathogen of swine that frequently causes serious outbreaks, in particular in Asia, and can also cause severe invasive disease in humans. In this study, 21 S. Choleraesuis isolates, detected from 21 patients with diarrhea in China between 2010 and 2011, were found to include 19 H2S-negative S. Choleraesuis isolates and two H2S-positive isolates. This is the first report of H2S-negative S. Choleraesuis isolated from humans. The majority of H2S-negative isolates exhibited high resistance to ampicillin, chloramphenicol, gentamicin, tetracycline, ticarcillin, and trimethoprim-sulfamethoxazole, but only six isolates were resistant to norfloxacin. In contrast, all of the isolates were sensitive to cephalosporins. Fifteen isolates were found to be multidrug resistant. In norfloxacin-resistant isolates, we detected mutations in the gyrA and parC genes and identified two new mutations in the parC gene. Pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and clustered regularly interspaced short palindromic repeat (CRISPR) analysis were employed to investigate the genetic relatedness of H2S-negative and H2S-positive S. Choleraesuis isolates. PFGE revealed two groups, with all 19 H2S-negative S. Choleraesuis isolates belonging to Group I and H2S-positive isolates belonging to Group II. By MLST analysis, the H2S-negative isolates were all found to belong to ST68 and H2S-positive isolates belong to ST145. By CRISPR analysis, no significant differences in CRISPR 1 were detected; however, one H2S-negative isolate was found to contain three new spacers in CRISPR 2. All 19 H2S-negative isolates also possessed a frame-shift mutation at position 760 of phsA gene compared with H2S-positive isolates, which may be responsible for the H2S-negative phenotype. Moreover, the 19 H2S-negative isolates have similar PFGE patterns and same mutation site in the phsA gene, these results indicated

  19. Antimicrobial Resistance and Molecular Investigation of H2S-Negative Salmonella enterica subsp. enterica serovar Choleraesuis Isolates in China.

    Directory of Open Access Journals (Sweden)

    Jing Xie

    Full Text Available Salmonella enterica subsp. enterica serovar Choleraesuis is a highly invasive pathogen of swine that frequently causes serious outbreaks, in particular in Asia, and can also cause severe invasive disease in humans. In this study, 21 S. Choleraesuis isolates, detected from 21 patients with diarrhea in China between 2010 and 2011, were found to include 19 H2S-negative S. Choleraesuis isolates and two H2S-positive isolates. This is the first report of H2S-negative S. Choleraesuis isolated from humans. The majority of H2S-negative isolates exhibited high resistance to ampicillin, chloramphenicol, gentamicin, tetracycline, ticarcillin, and trimethoprim-sulfamethoxazole, but only six isolates were resistant to norfloxacin. In contrast, all of the isolates were sensitive to cephalosporins. Fifteen isolates were found to be multidrug resistant. In norfloxacin-resistant isolates, we detected mutations in the gyrA and parC genes and identified two new mutations in the parC gene. Pulsed-field gel electrophoresis (PFGE, multilocus sequence typing (MLST, and clustered regularly interspaced short palindromic repeat (CRISPR analysis were employed to investigate the genetic relatedness of H2S-negative and H2S-positive S. Choleraesuis isolates. PFGE revealed two groups, with all 19 H2S-negative S. Choleraesuis isolates belonging to Group I and H2S-positive isolates belonging to Group II. By MLST analysis, the H2S-negative isolates were all found to belong to ST68 and H2S-positive isolates belong to ST145. By CRISPR analysis, no significant differences in CRISPR 1 were detected; however, one H2S-negative isolate was found to contain three new spacers in CRISPR 2. All 19 H2S-negative isolates also possessed a frame-shift mutation at position 760 of phsA gene compared with H2S-positive isolates, which may be responsible for the H2S-negative phenotype. Moreover, the 19 H2S-negative isolates have similar PFGE patterns and same mutation site in the phsA gene, these

  20. Genotypic Identification of AmpC β-Lactamases Production in Gram-Negative Bacilli Isolates.

    Science.gov (United States)

    Wassef, Mona; Behiry, Iman; Younan, Mariam; El Guindy, Nancy; Mostafa, Sally; Abada, Emad

    2014-01-01

    AmpC type β-lactamases are commonly isolated from extended-spectrum Cephalosporin-resistant Gram-negative bacteria. Also, resistance appeared in bacterial species not naturally producing AmpC enzymes. Therefore, a standard test for the detection of the plasmid-mediated AmpC enzyme and new breakpoints for extended spectrum Cephalosporins are urgently necessary. To detect plasmid and chromosomal mediated AmpC-β-lactamases in Gram negative bacteria in community and hospital acquired infections. 1073 Gram negative clinical isolates were identified by the conventional methods and were screened for AmpC production using Cefoxitin discs. Confirmatory phenotypic identifications were done for the Cefoxitin-resistant isolates using Boronic Acid for combined and double disc synergy tests, Cloxacillin based double disc synergy test, and induction tests. The genotypic identification of plasmid-mediated AmpC was done using multiplex PCR. ESBL production was also screened by discs of Ceftazidime and Cefotaxime with and without Clavulanic Acid (10 μg). The AmpC-producing isolates among all identified Gram negative bacilli were 5.8% (62/1073) as detected by screening disc diffusion methods, where 72% were positive for AmpC by combined disc method (Cefotetan and Boronic Acid), 56.5% were positive by each of Boronic Acid and Cloxacillin double disc synergy tests, 35.5% were positive by the induction test, and 25.8% were plasmid-mediated AmpC β-lactamase producers by the multiplex PCR. Plasmid-mediated AmpC genes retrieved, belonged to the families (MOX, FOX, EBC and CIT). ESBL producers were found in 26 (41.9%) isolates, 15 (57%) of which also produced AmpC. Isolates caused hospital acquired infections were (53/62); of which (39/62) were AmpC producers. While only (8/62) of the isolates caused community-acquired infections, were AmpC producers, and (1.6%) (1/62) were non AmpC producer. The AmpC β-lactamases detection tests had to be included in the routine microbiology workup of

  1. ESQUEMA SIMPLIFICADO PARA IDENTIFICAÇÃO DE ESTAFILOCOCOS COAGULASE-POSITIVOS ISOLADOS DE MASTITE BOVINA SIMPLIFIED SCHEME FOR IDENTIFICATION OF COAGULASE-POSITIVE STAPHYLOCOCCI ISOLATED FROM BOVINE MASTITIS

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    Maria Aparecida Vasconcelos Paiva Brito

    2002-02-01

    to bacitracin, aerobic acid from glycerol in the presence of 0.4mug m-1 of eritromicin, catalase, and coagulated rabbit plasma by the tube test. The acriflavin susceptibility was tested on Baird Parker and P agar plates containing 7mug m-1 of acriflavin. Five coagulase-negative S. hyicus strains identified by the API Staph system and S. aureus ATCC 29213 were used as test controls. Thirty-eight out of the 49 CPS were identified as S. aureus and 11 as S. hyicus, while S. intermedius was not found. The API Staph system was used to confirm the identification of the coagulase-positive strains of S. hyicus, seven S. aureus strains not producing acetoin and four negative for anaerobic fermentation of mannitol. All the strains identified as S. aureus were resistant to acriflavin whereas S. hyicus were sensitive. It was concluded that the sensitivity to acriflavin can be used in addition to the tests of coagulase and acetoin production to differentiate CPS isolated from bovine mastitis.

  2. Phenotypic and Genotypic Detection of Metallo-beta-lactamases among Imipenem-Resistant Gram Negative Isolates

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    Mohammad Mohammadzadeh

    2016-08-01

    Full Text Available Background:   Imipenem-resistant gram negative bacteria, resulting from metallo-beta-lactamase (MBLs-producing strains have been reported to be among the important causes of nosocomial infections and of serious therapeutic problem worldwide. Because of their broad range, potent carbapenemase activity and resistance to inhibitors, these enzymes can confer resistance to almost all beta-lactams. The prevalence of metallo-beta-lactamase among imipenem-resistant Acinetobacter spp., Pseudomonas spp. and Enerobacteriaceae isolates is determined.Methods:   In this descriptive study 864 clinical isolates of Acinetobacter spp., Pseudomonas spp. and Enterobacteriaceae, were initially tested for imipenem susceptibility. The metallo-beta-lactamase production was detected using combined disk diffusion, double disk synergy test, and Hodge test. Then all imipenem resistant isolates were tested by PCR for imp, vim and ndm genes. Results:   Among 864 isolates, 62 (7.17 % were imipenem-resistant. Positive phonetypic test for metallo-beta-lactamase was 40 (64.5%, of which 24 (17.1% and 16 (9.2% isolates were Acinetobacter spp. and Pseudomonas spp., respectively. By PCR method 30 (48.4% of imipenem resistant Acinetobacter, and Pseudomonas isolates were positive for MBL-producing genes. None of the Enterobacteriaceae isolates were positive for metallo-beta-lactamase activity. Conclusion:   The results of this study are indicative of the growing number of nosocomial infections associated with multidrug-resistant gram negative bacteria in this region leading to difficulties in antibiotic therapy. Thereby, using of phenotypic methods can be helpful for management of this problem.

  3. Characterization of staphylococci in urban wastewater treatment plants in Spain, with detection of methicillin resistant Staphylococcus aureus ST398.

    Science.gov (United States)

    Gómez, Paula; Lozano, Carmen; Benito, Daniel; Estepa, Vanesa; Tenorio, Carmen; Zarazaga, Myriam; Torres, Carmen

    2016-05-01

    The objective of this study was to determine the prevalence of Staphylococcus in urban wastewater treatment plants (UWTP) of La Rioja (Spain), and to characterize de obtained isolates. 16 wastewater samples (8 influent, 8 effluent) of six UWTPs were seeded on mannitol-salt-agar and oxacillin-resistance-screening-agar-base for staphylococci and methicillin-resistant Staphylococcus aureus recovery. Antimicrobial susceptibility profile was determined for 16 antibiotics and the presence of 35 antimicrobial resistance genes and 14 virulence genes by PCR. S. aureus was typed by spa, agr, and multilocus-sequence-typing, and the presence of immune-evasion-genes cluster was analyzed. Staphylococcus spp. were detected in 13 of 16 tested wastewater samples (81%), although the number of CFU/mL decreased after treatment. 40 staphylococci were recovered (1-5/sample), and 8 of them were identified as S. aureus being typed as (number of strains): spa-t011/agr-II/ST398 (1), spa-t002/agr-II/ST5 (2), spa-t3262/agr-II/ST5 (1), spa-t605/agr-II/ST126 (3), and spa-t878/agr-III/ST2849 (1). S. aureus ST398 strain was methicillin-resistant and showed a multidrug resistance phenotype. Virulence genes tst, etd, sea, sec, seg, sei, sem, sen, seo, and seu, were detected among S. aureus and only ST5 strains showed genes of immune evasion cluster. Thirty-two coagulase-negative Staphylococcus of 12 different species were recovered (number of strains): Staphylococcus equorum (7), Staphylococcus vitulinus (4), Staphylococcus lentus (4), Staphylococcus sciuri (4), Staphylococcus fleurettii (2), Staphylococcus haemolyticus (2), Staphylococcus hominis (2), Staphylococcus saprophyticus (2), Staphylococcus succinus (2), Staphylococcus capitis (1), Staphylococcus cohnii (1), and Staphylococcus epidermidis (1). Five presented a multidrug resistance phenotype. The following resistance and virulence genes were found: mecA, lnu(A), vga(A), tet(K), erm(C), msr(A)/(B), mph(C), tst, and sem. We found that

  4. AmpC beta lactamases among Gram negative clinical isolates from a tertiary hospital, South India

    OpenAIRE

    Parveen R. Mohamudha; Harish, B.N.; Parija, S.C.

    2010-01-01

    AmpC β-lactamases are cephalosporinases that hydrolyze cephamycins as well as other extended-spectrum cephalosporins and are poorly inhibited by clavulanic acid. Although reported with increasing frequency, the true rate of occurrence of AmpC β-lactamases in different organisms, including members of Enterobacteriaceae, remains unknown. The present study was designed to determine the occurrence of AmpC enzyme-harbouring Gram-negative clinical isolates in a tertiary care hospital in P...

  5. In vitro antimicrobial activity of five essential oils on multidrug resistant Gram-negative clinical isolates

    OpenAIRE

    Sakkas, Hercules; Gousia, Panagiota; Economou, Vangelis; Sakkas, Vassilios; Petsios, Stefanos; Papadopoulou, Chrissanthy

    2016-01-01

    Aim/Background: The emergence of drug-resistant pathogens has drawn attention on medicinal plants for potential antimicrobial properties. The objective of the present study was the investigation of the antimicrobial activity of five plant essential oils on multidrug resistant Gram-negative bacteria. Materials and Methods: Basil, chamomile blue, origanum, thyme, and tea tree oil were tested against clinical isolates of Acinetobacter baumannii (n = 6), Escherichia coli (n = 4), Klebsiella pneum...

  6. Conjunctival sac bacterial flora isolated prior to cataract surgery

    Directory of Open Access Journals (Sweden)

    Suto C

    2012-01-01

    Full Text Available Chikako Suto1,2, Masahiro Morinaga1,2, Tomoko Yagi1,2, Chieko Tsuji3, Hiroshi Toshida41Department of Ophthalmology, Saiseikai Kurihashi Hospital, Saitama; 2Department of Ophthalmology, Tokyo Women's Medical University, Tokyo; 3Department of Clinical Laboratory, Saiseikai Kurihashi Hospital, Saitama; 4Department of Ophthalmology, Juntendo University Shizuoka Hospital, Izunokuni, Shizuoka, JapanObjective: To determine the trends of conjunctival sac bacterial flora isolated from patients prior to cataract surgery.Subjects and methods: The study comprised 579 patients (579 eyes who underwent cataract surgery. Specimens were collected by lightly rubbing the inferior palpebral conjunctival sac with a sterile cotton swab 2 weeks before surgery, and then cultured for isolation of bacteria and antimicrobial sensitivity testing. The bacterial isolates and percentage of drug-resistant isolates were compared among age groups and according to whether or not patients had diabetes mellitus, hyperlipidemia, dialysis therapy, oral steroid use, dry eye syndrome, or allergic conjunctivitis.Results: The bacterial isolation rate was 39.2%. There were 191 strains of Gram-positive cocci, accounting for the majority of all isolates (67.0%, among which methicillin-sensitive coagulase-negative staphylococci was the most frequent (127 strains, 44.5%, followed by methicillin-resistant coagulase-negative staphylococci (37 strains, 12.7%. All 76 Gram-positive bacillary isolates (26.7% were from the genus Corynebacterium. Among the 16 Gram-negative bacillary isolates (5.9%, the most frequent was Escherichia coli (1.0%. The bacterial isolation rate was higher in patients >60 years old, and was lower in patients with dry eye syndrome, patients under topical treatment for other ocular disorders, and patients with hyperlipidemia. There was no significant difference in bacterial isolation rate with respect to the presence/absence of diabetes mellitus, steroid therapy, dialysis, or

  7. Aquaporin-4 antibody negative recurrent isolated optic neuritis: clinical evidence for disease heterogeneity.

    Science.gov (United States)

    Waschbisch, A; Atiya, M; Schaub, C; Derfuss, T; Schwab, S; Lee, D H; Müller, M; Linker, R A

    2013-08-15

    Recurrent optic neuritis is frequently observed in multiple sclerosis (MS) and is a typical finding in neuromyelitis optica (NMO). Patients that lack further evidence of demyelinating disease are diagnosed with RION (recurrent isolated optic neuritis) or CRION (chronic relapsing inflammatory neuropathy) if they require immunosuppressive therapy to prevent further relapses. The etiology and disease course of this rare condition are not well defined. We studied a series of 10 patients who presented with recurrent episodes of isolated optic neuritis (ON, n=57) and were followed over a median of 3.5 years. Visual acuity was severely reduced at the nadir of the disease (20/200 to 20/800). All patients had MRI non-diagnostic for MS/NMO and were aquaporin-4 antibody negative. Six patients fulfilled the CRION criteria. In two of these a single ON followed by a long disease-free interval preceded development of CRION for years, suggesting the conversion of an initially "benign" isolated ON into the chronic relapsing course. Cerebrospinal fluid (CSF) analysis revealed mild pleocytosis in 5 patients, identical oligoclonal bands in serum and CSF were observed in 2 patients, while the others remained negative. In conclusion, recurrent ON is a disease entity that requires aggressive glucocorticoid and eventually long-term immunosuppressive therapy to prevent substantial visual impairment.

  8. Phenotypic and genotypic antimicrobial resistance of staphylococci from bovine milk.

    Science.gov (United States)

    Kot, B; Piechota, M; Wolska, K M; Frankowska, A; Zdunek, E; Binek, T; Kłopotowska, E; Antosiewicz, M

    2012-01-01

    The aim of this study was to examine phenotypic and genotypic antimicrobial resistance of staphylococci from milk samples from cows with subclinical and clinical mastitis and from cows without mastitis symptoms to methicillin, tetracyclines, macrolides and lincosamides (ML). Of 207 strains, including 34 S. aureus and 173 coagulase-negative staphylococci (CNS), 11 (6.4%) CNS strains were phenotypically resistant to methicillin. The mecA gene was detected by PCR only in two S. xylosus strains and one strain of S. epidermidis and S. simulans. No methicillin-resistant S. aureus strains were observed. In methicillin-resistant strains with mecA, gene resistance to other investigated antibiotics was not observed. Phenotypic resistance to tetracycline was detected in 11.0% of CNS strains and 47.4% of them carried the tetK gene. Of 173 CNS strains studied, 27 (15.6%) were resistant to at least one ML antibiotic. The resistance gene ermC was detected in 55.5% of the 27 ML-resistant strains. The ermA and ermB genes were detected in 14.8% and 11.1% of ML-resistant CNS strains, respectively. Antimicrobial resistance to methicillin, tetracyclines and macrolides was detected more frequently in staphylococcal strains from clinical mastitis compared to animals with subclinical symptoms and without mastitis, while the resistance to lincosamides showed a similar frequency in all groups of cows. In conclusion, CNS species from bovine milk differ in phenotypic and genotypic antimicrobial resistance profiles, and the use of PCR technique alone for the detection of methicillin, macrolide, lincosamide and tetyracycline resistance in CNS from cattle is not reliable.

  9. A vibration isolation system in low frequency excitation region using negative stiffness structure for vehicle seat

    Science.gov (United States)

    Le, Thanh Danh; Ahn, Kyoung Kwan

    2011-12-01

    This paper designs and fabricates a vibration isolation model for improving vibration isolation effectiveness of the vehicle seat under low excitation frequencies. The feature of the proposed system is to use two symmetric negative stiffness structures (NSS) in parallel to a positive stiffness structure. Here, theoretical analysis of the proposed system is clearly presented. Then, the design procedure is derived so that the resonance peak of frequency-response curve drifts to the left, the load support capacity of the system is maintained, the total size of the system is reduced for easy practical application and especially, the bending of the frequency-response curve is minimized. Next the dynamic equation of the proposed system is set up. Then, the harmonic balance (HB) method is employed to seek the characteristic of the motion transmissibility of the proposed system at the steady state for each of the excitation frequency. From this characteristic, the curves of the motion transmission are predicted according to the various values of the configurative parameters of the system. Then, the time responses to the sinusoidal, multi frequency and random excitations are also investigated by simulation and experiment. In addition, the isolation performance comparison between the system with NSS and system without NSS is realized. The simulation results reveal that the proposed system has larger frequency region of isolation than that of the system without NSS. The experimental results confirm also that with a random excitation mainly spreading from 0.1 to 10 Hz, the isolation performance of the system with NSS is greatly improved, where the RMS values of the mass displacement may be reduced to 67.2%, whereas the isolation performance of the system without NSS is bad. Besides, the stability of the steady-state response is also studied. Finally, some conclusions are given.

  10. Resistance to antibiotics in Gram-negative bacteria isolated from broiler carcasses

    Directory of Open Access Journals (Sweden)

    Moreira M.A.S.

    2002-01-01

    Full Text Available One hundred and ninety-seven isolates of Gram-negative bacteria, comprising 10 genera, were isolated from poultry carcasses at a processing plant in order to investigate resistance to low levels of antibiotics. The samples were taken just after evisceration and before inspection. Most of the isolates were of Samonella and Escherichia. Other genera present were Enterobacter, Serratia, Klebsiella, Kluyvera, Erwinia, Citrobacter, Pseudomonas and Aeromonas. Distinct profiles of antibiotic resistance were detected. Resistance to more than two antibiotics predominated and spanned several classes of antibiotics. Salmonellae and escherichiae were mainly resistant to the aminoglycosides, followed by tetracycline, nitrofuran, sulpha, macrolide, chloramphenicol, quinolones and beta-lactams. Most isolates were sensitive to 30mug/ml olaquindox, the growth promoter in use at the time of sampling. However, many were resistant to a level of 10mug/ml and 13mug/ml olaquindox, levels present in the gut due to the dilution in the feed. The results suggest a possible role of low level administration of antibiotics to broilers in selecting multi-resistant bacteria in vivo.

  11. Catalase-negative Staphylococcus aureus isolated from a diabetic foot ulcer

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    MR Zali

    2010-12-01

    Full Text Available We report a catalase-negative Staphylococcus aureus isolated from a 56-year-old male diabetic patient with foot ulcer who attended our surgery ward. Species identification was confirmed by Gram staining, standard biochemical tests and PCR amplification of the nuc and fem genes. Antibiotic susceptibility showed that the strain was sensitive to imepenem, chloramphenicol, amoxicillin, vancomycin and resistant to oxacillin, penicillin, ceftriaxone, erythromycin, clindamycin, and amikacin. Clinicians and microbiologists must be encouraged to identify and report these atypical strains and the infections associated with them in order to establish their role in pathogenesis.

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

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    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.

  13. NEGATIVE EFFECTS OF ETHANOL ON ISOLATED GUINEA PIG LEFT ATRIUM AND PAPILLARY MUSCLES

    Institute of Scientific and Technical Information of China (English)

    胡浩; 臧伟进; 于晓江; 王昌利; 张凤杰; 孙强; 张春虹

    2002-01-01

    Objective To investigate the effects of ethanol on physiologic al characteristics of the isolated guinea pig left atrium and papillary muscles.Methods The effects of ethanol on contractility, post-rest potentiatio n and positive staircase phenomenon were observed in isolated left atrium and pa pillary muscles of guinea pig.Results Ethanol(50.0,100.0,200.0mmol*L-1)prominently inhibited the con traction of papillary muscles. Ethanol(12.5,25.0,50.0,100.0,200.0mmol*L-1 ) inhibited the contraction of left atrium, and markedly decreased the post-r est potentiation of myocardial contractility in left atrium. High concentration of ethanol(100,200mmol*L-1) depressed the positive staircase phenomenon of isolated guinea pig left atrium.Conclusion These results suggest that ethanol induces inhibitor y effects of the contractility, post-rest potentiation, positive staircase phen omenon of left atrium. The mechanism by which ethanol induces the negative inotr opic effects may be related to decrease the amount of calcium released from the intracellular stores.

  14. NEGATIVE EFFECTS OF ETHANOL ON ISOLATED GUINEA PIG LEFT ATRIUM AND PAPILLARY MUSCLES

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Objective :To investigate the effects of ethanol on physiological characteristics of the isolated guinea pig left atrium and papillary muscles,Methods:The effects of ethanol on contractility,post-rest potentiation and positive staircase phenomenon were observed in isolated left atrium and papillary muscles of guinea pig.Results:Ethanol(50.0,100.0,200.0mmol.L-1) prominently inhibited the contraction of papillary muscles.Ethanol(12.5,25.0,50.0,100.0,200.0mmol.L-1)inhibited the contraction of left atrium,and markedly decreased the post-rest potentiation of myocardial contractility in left atrium.High concentration of ethanol(100,200mmol.L-1)depressed the positive staircase phenomenon of isolated guinea pig left atrium.Conclusion:These results suggest that ethanol induces inhibitory effects of the contractility,post-rest potentiation,positive staircase phenomenon of letf atrium.The mechanism by which ethanol induces the negative inotropic effects may be related to decrease the amount of calcium released from the intracellular stores.

  15. Identification and enumeration of staphylococci from the eye during soft contact lens wear.

    Science.gov (United States)

    Leitch, E C; Harmis, N Y; Corrigan, K M; Willcox, M D

    1998-04-01

    An extensive study was conducted to identify and enumerate staphylococcal microbiota found on ocular sites during asymptomatic soft contact lens (SCL) wear. A biochemical identification system separately grouped the eight clinically relevant staphylococci. Total counts and isolation frequencies from SCLs and ocular sites were evaluated. The epidermidis group was the most numerous isolate from extended wear (EW) lenses; the capitis/warneri group was the most numerous (p microbiota of the ocular surface during SCL wear. Furthermore, an increase and a shift in staphylococcal microbiota between DW and EW regimes was highlighted.

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

    Directory of Open Access Journals (Sweden)

    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.

  17. Antimicrobial Activity of Carbon Nanoparticles Isolated from Natural Sources against Pathogenic Gram-Negative and Gram-Positive Bacteria

    Directory of Open Access Journals (Sweden)

    Sheena Varghese

    2013-01-01

    Full Text Available This paper describes the isolation of carbon nanoparticles (CNPs from kitchen soot, characterization of the CNPs by UV/visible spectroscopy, SEM and XRD, and their antimicrobial action. The antibacterial activity of the isolated carbon nanoparticles was tested against various pathogenic bacterial strains such as Gram-negative Proteus refrigere and Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus and Streptococcus haemolyticus. The inhibition zones were measured, and it was found that the carbon nanoparticles isolated from natural sources are active against these Gram-negative and Gram-positive bacterial strains.

  18. Identification and antimicrobial susceptibility patterns of Staphylococcus spp. isolated from canine chronic otitis externa

    Directory of Open Access Journals (Sweden)

    Silva N.

    2001-01-01

    Full Text Available Swab samples obtained from 96 dogs with chronic otitis externa were cultured for the isolation of Staphylococcus species. Of 57 staphylococcal strains, 41 (72% were coagulase-negative (CNS. The identification of staphylococci strains was made by standard procedures for the routine identification of staphylococci in clinical practice. S. sciuri was the most frequent species isolated (22.8% from chronic otitis externa in dogs followed by S. intermedius (12.3%, S. auricularis (10.5% and S. aureus (8.8%. Three (5.2% CNS strains could not be identified. Bacterial isolates were susceptible to enrofloxacin, gentamicin, cephalothin, chloramphenicol and neomycin. Resistance was most common to penicillin G, oxacillin and ampicillin.

  19. Correction: Membrane-active macromolecules resensitize NDM-1 gram-negative clinical isolates to tetracycline antibiotics.

    Directory of Open Access Journals (Sweden)

    Divakara S S M Uppu

    Full Text Available Gram-negative 'superbugs' such as New Delhi metallo-beta-lactamase-1 (blaNDM-1 producing pathogens have become world's major public health threats. Development of molecular strategies that can rehabilitate the 'old antibiotics' and halt the antibiotic resistance is a promising approach to target them. We report membrane-active macromolecules (MAMsthat restore the antibacterial efficacy (enhancement by >80-1250 fold of tetracycline antibiotics towards blaNDM-1 Klebsiella pneumonia and blaNDM-1 Escherichia coli clinical isolates.Organismic studies showed that bacteria had an increased and faster uptake of tetracyclinein the presence of MAMs which is attributed to the mechanism of re-sensitization. Moreover,bacteria did not develop resistance to MAMs and MAMs stalled the development of bacterial resistance to tetracycline. MAMs displayed membrane-active properties such as dissipation of membrane potential and membrane-permeabilization that enabled higher uptake of tetracycline in bacteria. In-vivo toxicity studies displayed good safety profiles and preliminary in-vivo antibacterial efficacy studies showed that mice treated with MAMs in combination with antibiotics had significantly decreased bacterial burden compared to the untreated mice. This report of re-instating the efficacy of the antibiotics towards blaNDM-1 pathogens using membrane-active molecules advocates their potential for synergistic co-delivery of antibiotics to combat Gram-negative superbugs.

  20. Membrane-active macromolecules resensitize NDM-1 gram-negative clinical isolates to tetracycline antibiotics.

    Directory of Open Access Journals (Sweden)

    Divakara S S M Uppu

    Full Text Available Gram-negative 'superbugs' such as New Delhi metallo-beta-lactamase-1 (blaNDM-1 producing pathogens have become world's major public health threats. Development of molecular strategies that can rehabilitate the 'old antibiotics' and halt the antibiotic resistance is a promising approach to target them. We report membrane-active macromolecules (MAMs that restore the antibacterial efficacy (enhancement by >80-1250 fold of tetracycline antibiotics towards blaNDM-1 Klebsiella pneumonia and blaNDM-1 Escherichia coli clinical isolates. Organismic studies showed that bacteria had an increased and faster uptake of tetracycline in the presence of MAMs which is attributed to the mechanism of re-sensitization. Moreover, bacteria did not develop resistance to MAMs and MAMs stalled the development of bacterial resistance to tetracycline. MAMs displayed membrane-active properties such as dissipation of membrane potential and membrane-permeabilization that enabled higher uptake of tetracycline in bacteria. In-vivo toxicity studies displayed good safety profiles and preliminary in-vivo antibacterial efficacy studies showed that mice treated with MAMs in combination with antibiotics had significantly decreased bacterial burden compared to the untreated mice. This report of re-instating the efficacy of the antibiotics towards blaNDM-1 pathogens using membrane-active molecules advocates their potential for synergistic co-delivery of antibiotics to combat Gram-negative superbugs.

  1. Highly frequent mutations in negative regulators of multiple virulence genes in group A streptococcal toxic shock syndrome isolates.

    Science.gov (United States)

    Ikebe, Tadayoshi; Ato, Manabu; Matsumura, Takayuki; Hasegawa, Hideki; Sata, Tetsutaro; Kobayashi, Kazuo; Watanabe, Haruo

    2010-04-01

    Streptococcal toxic shock syndrome (STSS) is a severe invasive infection characterized by the sudden onset of shock and multiorgan failure; it has a high mortality rate. Although a number of studies have attempted to determine the crucial factors behind the onset of STSS, the responsible genes in group A Streptococcus have not been clarified. We previously reported that mutations of csrS/csrR genes, a two-component negative regulator system for multiple virulence genes of Streptococcus pyogenes, are found among the isolates from STSS patients. In the present study, mutations of another negative regulator, rgg, were also found in clinical isolates of STSS patients. The rgg mutants from STSS clinical isolates enhanced lethality and impaired various organs in the mouse models, similar to the csrS mutants, and precluded their being killed by human neutrophils, mainly due to an overproduction of SLO. When we assessed the mutation frequency of csrS, csrR, and rgg genes among S. pyogenes isolates from STSS (164 isolates) and non-invasive infections (59 isolates), 57.3% of the STSS isolates had mutations of one or more genes among three genes, while isolates from patients with non-invasive disease had significantly fewer mutations in these genes (1.7%). The results of the present study suggest that mutations in the negative regulators csrS/csrR and rgg of S. pyogenes are crucial factors in the pathogenesis of STSS, as they lead to the overproduction of multiple virulence factors.

  2. IDENTIFICATION OF ENTEROTOXIGENIC STAPHYLOCOCCI IN MEAT RAW MATERIALS

    Directory of Open Access Journals (Sweden)

    D. S. Bataeva

    2016-01-01

    Full Text Available The need to renew the safety criteria for meat raw material used in production of fermented sausages was determined. The presence of the enterotoxigenic S. aureus strains in meat raw material was established. The method was mastered and the PCR approaches to identification and screenings of enterotoxigenic strains and their toxins in meat raw material were proposed. In order to identify enterotoxigenicity of detected staphylococci, the conservative regions of the S. aureus target gene sequences responsible for production of different types of enterotoxins (A, B, Е, С, D were found and studied. In addition, short fragments of a nuclear acid (primers corresponding to these revealed genes were constructed. As a result of identification, it was established that two isolated strains of S. аureus were enterotoxigenic. One of them produced type А and Е toxins (strain NG1 and another produced type С and Е toxins (strain NG2. The sensitivity and specificity of the real-time PCR method allowed not only identification of pure cultures but also screening of enterotoxigenic strains and their toxins in a product. The use of this method in the production control for the presence of the enterotoxigenic strains of S. aureus in meat raw material used in fermented sausage manufacture was recommended.

  3. Isolation of methicillin-resistant Staphylococcus spp. from ready-to-eat fish products.

    Science.gov (United States)

    Sergelidis, D; Abrahim, A; Papadopoulos, T; Soultos, N; Martziou, E; Koulourida, V; Govaris, A; Pexara, A; Zdragas, A; Papa, A

    2014-11-01

    A hundred samples from ready-to-eat (RTE) fish products were examined for the presence and antimicrobial susceptibility of Staphylococcus spp. Staphylococci were isolated from 43% of these samples (n = 100). The identified species in the samples were Staphylococcus aureus (7%), Staphylococcus epidermidis (13%), Staphylococcus xylosus (12%), Staphylococcus sciuri (4%), Staphylococcus warneri (3%), Staphylococcus saprophyticus (2%), Staphylococcus schleiferi (1%) and Staphylococcus auricularis (1%). Two Staph. aureus (MRSA) isolates, three Staph. epidermidis (MRSE), five Staph. xylosus, four Staph. sciuri, one Staph. schleiferi and one Staph. saprophyticus isolates were resistant to oxacillin and all of them carried the mecA gene. The two MRSA isolates belonged to the spa types t316 (ST359) and t548 (ST5) and none of them was able to produce enterotoxins. Pulsed field gel electrophoresis for Staph. aureus and Staph. epidermidis isolates revealed 6 and 11 distinct PFGE types, respectively, reflecting diversity. The presence of methicillin-resistant staphylococci, especially MRSA and MRSE, in RTE fish products may constitute a potential health risk for consumers. This study provides the first data on the occurrence of methicillin-resistant Staphylococcus aureus and methicillin-resistant coagulase-negative staphylococci in salted and smoked fish products in Greece. These results are important and useful for Staphylococcus spp. risk assessment and management programmes for ready-to-eat fish products. © 2014 The Society for Applied Microbiology.

  4. β-Lactamase production in key gram-negative pathogen isolates from the Arabian Peninsula.

    Science.gov (United States)

    Zowawi, Hosam M; Balkhy, Hanan H; Walsh, Timothy R; Paterson, David L

    2013-07-01

    SUMMARY Infections due to Gram-negative bacilli (GNB) are a leading cause of morbidity and mortality worldwide. The extent of antibiotic resistance in GNB in countries of the Gulf Cooperation Council (GCC), namely, Saudi Arabia, United Arab Emirates, Kuwait, Qatar, Oman, and Bahrain, has not been previously reviewed. These countries share a high prevalence of extended-spectrum-β-lactamase (ESBL)- and carbapenemase-producing GNB, most of which are associated with nosocomial infections. Well-known and widespread β-lactamases genes (such as those for CTX-M-15, OXA-48, and NDM-1) have found their way into isolates from the GCC states. However, less common and unique enzymes have also been identified. These include PER-7, GES-11, and PME-1. Several potential risk factors unique to the GCC states may have contributed to the emergence and spread of β-lactamases, including the unnecessary use of antibiotics and the large population of migrant workers, particularly from the Indian subcontinent. It is clear that active surveillance of antimicrobial resistance in the GCC states is urgently needed to address regional interventions that can contain the antimicrobial resistance issue.

  5. Countermeasures to Mitigate the Negative Impact of Sensory Deprivation and Social Isolation in Long-Duration Space Flight

    Science.gov (United States)

    Bachman, Katharine Ridgeway OBrien; Otto, Christian; Leveton, Lauren

    2012-01-01

    Long-duration space flight presents several challenges to the behavioral health of crew members. The environment that they are likely to experience will be isolated, confined, and extreme (ICE) and, as such, crew members will experience extreme sensory deprivation and social isolation. The current paper briefly notes the behavioral, cognitive, and affective consequences of psychological stress induced by ICE environments and proposes nine countermeasures aimed at mitigating the negative effects of sensory deprivation and social isolation. Implementation of countermeasures aims to maintain successful crew performance and psychological well-being in a long-duration space flight mission.

  6. Antimicrobial resistance pattern of Gram –negative bacilli isolated of Vali-Asr Hospital wards in Arak

    Directory of Open Access Journals (Sweden)

    Farshid Didgar

    2014-11-01

    Full Text Available Background: Infectious diseases are of the most important causes of mortality all around the world particular in developing countries. Recently, the most important thing that has worried medical society is antibiotic resistance. Multi-resistant gram_negative rods are important pathogens in hospitals, causing high rate of mortality.The main goal of this study was to investigate the antimicrobial resistance patterns among common gram-negative bacilli isolated from patients of Vali-Asr Hospital. Material and Methods: This is a cross-sectional descriptive study conducted between the years 2010-2012 in Vali-Asr hospital in Arak. In this study 1120 specimen were examined. Bacterial strains were isolated by conventional methods from various clinical samples of patients including: blood, urine, wound, sputum, CSF, andetc.All isolates were examined for antimicrobial resistance using disc diffusion method. Results: In this study 737 specimen were positive cultures. A total of 332 isolates of Gram-negative bacilli were identified. The most frequent gram negative bacteria were isolated from urine, wound, blood, respiratory secretion and catheter. The most frequent pathogens were E.coli followed by k.pneumonia, entrobacter, p.oaeruginosa, Acinetobacter spp, citrobacter and proteus. High rate of resistance to third generation of cephalospoins & carbapenems observed amang isolates of Acintobacter spp.Prodution of extended spectrum beralactamases (ESBLS was found in 51.4% of all Gram negative bacteria. Conclusion: Antibiotic resistance, particularly multi-drug resistance is frequent among microorganisms of ValiAsr Hospital. Resistance in our country, like other countries have been shown to be increased, so it is highly recommended to prohibit unnecessary prescription of antibiotics.

  7. Simplest identification, O-specific polysaccharide purification and antigenic evaluation of Salmonella enterica serovar Typhi Vi negative isolate.

    Science.gov (United States)

    Salman, Muhammad; Ali, Aamir; Jabbar, Abdul; Sarwar, Yasra; Rahman, Moazur; Iqbal, Mazhar; Haque, Abdul

    2015-01-01

    Currently licensed typhoid vaccines are based on Vi capsular polysaccharides. Recent molecular reports from typhoid endemic countries state that Salmonella enterica serovar Typhi (S. Typhi) Vi negative strains occur naturally and cause typhoid fever which is indistinguishable from disease caused by Vi positive strains. Vaccine based on Vi polysaccharide may not protect patients if the invading S. Typhi are negative for Vi. The lipopolysaccharide (LPS) is an essential component of S. Typhi outer membrane in which O-specific polysaccharide (OSP) is a protective antigen and universal candidate for vaccine development. In this study, S. Typhi Vi negative isolates were discriminated from Vi positive isolates through a duplex PCR using primers of fliC-d (599bp) and tviA (495bp) genes. The LPS of S. Typhi Vi negative isolates was extracted by hot phenol method and OSP was purified by core hydrolysis. The yield of extracted LPS was 91 mg/L and that of purified OSP was 49.14 mg/L of culture broth. LPS showed ladder like appearance by zinc imidazole staining following SDS-PAGE. Whole cell challenged mice sera were used for in vitro antigenicity evaluation of the purified LPS and OSP. The antigenicity was found adequate by immunodiffusion assay. To our knowledge, this is the first report of purification and antigenic evaluation of LPS of a Vi negative S. Typhi isolate. The purified OSP from S. Typhi Vi negative isolate may be coupled with a carrier protein to produce universal low cost conjugate vaccine candidates for use in typhoid endemic regions.

  8. Protein-based biofilm matrices in Staphylococci

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    Pietro eSpeziale

    2014-12-01

    Full Text Available Staphylococcus aureus and Staphylococcus epidermidis are the most important etiological agents of biofilm associated-infections on indwelling medical devices. Biofilm infections may also develop independently of indwelling devices, e.g. in native valve endocarditis, bone tissue and open wounds. After attachment to tissue or indwelling medical devices that have been conditioned with host plasma proteins, staphylococcal biofilms grow and produce a specific environment which provides the conditions for cell-cell interaction and formation of multicellular communities. Bacteria living in biofilms express a variety of macromolecules, including exopolysaccharides, proteins, extracellular eDNA and other polymers. The S. aureus surface protein C and G (SasC and SasG, clumping factor B (ClfB, serine aspartate repeat protein (SdrC, the biofilm-associated protein (Bap and the fibronectin/fibrinogen-binding proteins (FnBPA and FnBPB are individually implicated in biofilm matrix formation. In S. epidermidis, a protein named accumulation-associated protein (Aap contributes to both the primary attachment phase and the establishment of intercellular connections by forming fibrils on the cell surface. In S. epidermidis proteinaceous biofilm formation can also be mediated by the extracellular matrix binding protein (Embp and S. epidermidis surface protein C (SesC. Additionally, multifunctional proteins such as extracellular adherence protein (Eap and extracellular matrix protein binding protein (Emp of S. aureus and the iron-regulated surface determinant protein C (IsdC of S. lugdunensis can promote biofilm formation in iron-depleted conditions. This multitude of proteins intervene at different stages of biofilm formation with certain proteins contributing to biofilm accumulation and others mediating primary attachment to surfaces. This review examines the contribution of proteins to biofilm formation in staphylococci. The potential to develop vaccines to prevent

  9. Highly frequent mutations in negative regulators of multiple virulence genes in group A streptococcal toxic shock syndrome isolates.

    Directory of Open Access Journals (Sweden)

    Tadayoshi Ikebe

    2010-04-01

    Full Text Available Streptococcal toxic shock syndrome (STSS is a severe invasive infection characterized by the sudden onset of shock and multiorgan failure; it has a high mortality rate. Although a number of studies have attempted to determine the crucial factors behind the onset of STSS, the responsible genes in group A Streptococcus have not been clarified. We previously reported that mutations of csrS/csrR genes, a two-component negative regulator system for multiple virulence genes of Streptococcus pyogenes, are found among the isolates from STSS patients. In the present study, mutations of another negative regulator, rgg, were also found in clinical isolates of STSS patients. The rgg mutants from STSS clinical isolates enhanced lethality and impaired various organs in the mouse models, similar to the csrS mutants, and precluded their being killed by human neutrophils, mainly due to an overproduction of SLO. When we assessed the mutation frequency of csrS, csrR, and rgg genes among S. pyogenes isolates from STSS (164 isolates and non-invasive infections (59 isolates, 57.3% of the STSS isolates had mutations of one or more genes among three genes, while isolates from patients with non-invasive disease had significantly fewer mutations in these genes (1.7%. The results of the present study suggest that mutations in the negative regulators csrS/csrR and rgg of S. pyogenes are crucial factors in the pathogenesis of STSS, as they lead to the overproduction of multiple virulence factors.

  10. Lymph Node Negative Colorectal Cancers with Isolated Tumor Deposits Should Be Classified and Treated As Stage III

    NARCIS (Netherlands)

    Belt, E.J.T.; van Stijn, M.F.M.; Bril, H.; de Lange-de Klerk, E.S.M.; Meijer, G.A.; Meijer, S.; Stockmann, H.B.A.C.

    2010-01-01

    BACKGROUND: The prognostic role of pericolic or perirectal isolated tumor deposits (ITDs) in node-negative colorectal cancer (CRC) patients is unclear. Rules to define ITDs as regional lymph node metastases changed in subsequent editions of the TNM staging without substantial evidence. Aim of this

  11. Occurrence of false positive results for the detection of carbapenemases in carbapenemase-negative Escherichia coli and Klebsiella pneumoniae isolates.

    Directory of Open Access Journals (Sweden)

    Peng Wang

    Full Text Available Adequate detection of the production of carbapenemase in Enterobacteriaceae isolates is crucial for infection control measures and the appropriate choice of antimicrobial therapy. In this study, we investigated the frequency of false positive results for the detection of carbapenemases in carbapenemase-negative Escherichia coli and Klebsiella pneumoniae clinical isolates by the modified Hodge test (MHT. Three hundred and one E. coli and K. pneumoniae clinical isolates were investigated. All produced extended spectrum β-lactamases (ESBLs but were susceptible to carbapenems. Antimicrobial susceptibility testing was performed by the disk diffusion and agar dilution methods. The MHT was performed using the standard inoculum of test organisms recommended by the CLSI. Genes that encoded ESBLs and carbapenemases were identified by PCR and DNA sequencing. Among the 301 clinical isolates, none of the isolates conformed to the criteria for carbapenemase screening recommended by the CLSI. The susceptibility rates for imipenem, meropenem, and ertapenem all were 100.0%, 100.0%, and 100.0%, respectively. Of the 301 E. coli and K. pneumoniae isolates, none produced carbapenemase. The MHT gave a positive result for 3.3% (10/301 of the isolates. False positive results can occur when the MHT is used to detect carbapenemase in ESBL-producing isolates and clinical laboratories must be aware of this fact.

  12. Simulated and experimental studies on a high-static-low-dynamic stiffness isolator using magnetic negative stiffness spring

    Science.gov (United States)

    Dong, Guangxu; Zhang, Xinong; Xie, Shilin; Yan, Bo; Luo, Yajun

    2017-03-01

    For the purpose of isolating the low frequency vibration, a magnetic vibration isolator with the feature of high-static-low-dynamic stiffness (HSLDS) is developed in this paper, which is constructed by combining a magnetic negative stiffness spring (MNSS) with a spiral flexure spring (SFS) in parallel. The MNSS comprises three magnetic rings configured in attraction and is utilized to reduce the resonant frequency of the isolator. Then an analytical expression of magnetic negative stiffness (MNS) of the MNSS is deduced in terms of the current model, and an approximation to the MNS is further sought. To support the object, the axial positive stiffness of SFSs, which can behave with a smaller static deformation if a specified weight is applied, is analyzed with finite element method (FEM). After that, the governing equation of the isolator is established and solved via harmonic balance method (HBM). Finally, an experimental prototype is developed and tested. The experimental results demonstrate that the MNSS can reduce the resonant frequency of the isolator to expand the isolation frequency band to low frequency range; and the theoretical calculations and experimental results shows a good agreement.

  13. PSM-Mec - A Virulence Determinant that Connects TranscriptionalRegulation, Virulence, and Antibiotic Resistance in Staphylococci

    OpenAIRE

    Li Qin; Joshua McCausland; Gordon Cheung; Michael Otto

    2016-01-01

    PSM-mec is a secreted virulence factor that belongs to the phenol-soluble modulin (PSM) family of amphipathic, alpha-helical peptide toxins produced by Staphylococcus species. All known PSMs are core genome-encoded with the exception of PSM-mec, whose gene is found in specific sub-types of SCCmec methicillin resistance mobile genetic elements present in methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci. In addition to the cytolytic translational product, PSM-mec...

  14. [In vitro activity of trovafloxacin, of other fluoroquinolones and of related antimicrobials against clinical isolates. Grupo colaborativo WHONET-Argentina].

    Science.gov (United States)

    Rossi, A; Galas, M; Tokumoto, M; Guelfand, L; Lopardo, H

    1999-01-01

    The in vitro activity of trovafloxacin (TRV) has been evaluated in comparison with that of other antimicrobial agents against 5671 clinical isolates recovered by representative institutions of different provinces in our country. The resistance percentage to gentamicin and third generation cephalosporins among enterobacteriaceae was high: 17% and 16% respectively, with a considerable variation according to the analyzed species. The resistance to ciprofloxacin (CIP) and TRV affected approximately 9% of the isolates, without significant differences between both drugs. Fluoroquinolones (FQ) presented excellent activity on 166 isolates of Salmonella spp., 208 of Shigella flexneri and 76 of Shigella sonnei, where only one S.sonnei isolate was resistant to CIP, but susceptible to TRV. About half the isolates of Salmonella spp. and S.sonnei and almost all S.flexneri isolates were resistant to ampicillin, and more than 60% of Shigella spp. isolates were resistant to trimethoprim-sulfamethoxazole. A 41% of Staphylococcus aureus and 55% of coagulase-negative staphylococci isolates were resistant to oxacillin, presenting a highly associated multi-resistance. The resistance to FQ was also strongly related to oxacillin resistance, but the resistance to TRV was significantly lower than the CIP resistance: 9% vs 57% for S.aureus and 4% vs 41% for coagulase-negative staphylococci. A similar behavior was observed with Enterococcus spp., where 54% of the isolates were resistant to norfloxacin and only 13% were resistant to TRV. Neither Streptococcus pneumoniae (n = 193) nor Haemophilus influenzae (n = 139) isolates presented resistant to TRV.

  15. Phenotypic and molecular assessment of antimicrobial resistance profile of airborne Staphylococcus spp. isolated from flats in Kraków.

    Science.gov (United States)

    Lenart-Boroń, Anna; Wolny-Koładka, Katarzyna; Juraszek, Katarzyna; Kasprowicz, Andrzej

    2017-01-01

    Bacteria of the genus Staphylococcus were isolated from air sampled from living spaces in Kraków (Poland). In total, 55 strains belonging to the genus Staphylococcus were isolated from 45 sites, and 13 species of coagulase-negative staphylococci were identified. The species composition of studied airborne microbiota contains Staphylococcus species that are rarely infectious to humans. Most commonly isolated species comprised S. hominis and S. warneri. The disk-diffusion tests showed that the collected isolates were most frequently resistant to erythromycin. The PCR technique was employed to search for genes conferring the resistance in staphylococci to antibiotics from the group of macrolides, lincosamides and streptogramins. The analyzed Staphylococcus isolates possessed simultaneously 4 different resistance genes. The molecular analysis with the use of specific primers allowed to determine the most prevalent gene which is mphC, responsible for the resistance to macrolides and for the enzymatic inactivation of the drug by phosphotransferase. The second most often detected gene was msrA1, which confers the resistance of staphylococci to macrolides and is responsible for active pumping of antimicrobial particles out of bacterial cells.

  16. Molecular detection of ESBLs production and antibiotic resistance patterns in Gram negative bacilli isolated from urinary tract infections.

    Science.gov (United States)

    Kargar, Mehdi; Kargar, Mohammad; Jahromi, Mohammad Zareian; Najafi, Akram; Ghorbani-Dalini, Sadegh

    2014-01-01

    β-lactam resistance is more prevalent in Gram negative bacterial isolates worldwide, particularly in developing countries. In order to provide data relating to antibiotic therapy and resistance control, routine monitoring of corresponding antibiotic resistance genes is necessary. The aim of this study was the characterization of β-lactam resistance genes and its plasmid profile in bacteria isolated from urinary tract infection samples. In this study, 298 Gram negative bacteria isolated from 6739 urine specimens were identified by biochemical standard tests. Antimicrobial susceptibility testing was performed by the disk diffusion method. Extended-spectrum β-lactamase (ESBL)-producing strains were also detected by the double-disk synergy test. The presence of blaTEM and blaSHV genes in the strains studied was ascertained by polymerase chain reaction. Of all Gram negative bacteria, Escherichia coli (69.1%) was the most common strain, followed by Klebsiella sp. (12.1%), Enterobacter sp. (8.4%), Proteus sp. (4.4%), Citrobacter (4%) and Pseudomonas sp. (2%). The most antibiotic resistance was shown to tetracycline (95.16%), nalidixic acid (89.78%) and gentamycin (73.20%) antibiotics. Among all the strains tested, 35 isolates (11.75%) expressed ESBL activity. The prevalence of TEM and SHV positivity among these isolates was 34.29%, followed by TEM (31.43%), TEM and SHV negativity (20.0%) and SHV (14.29%), respectively. Regular monitoring of antimicrobial drug resistance seems necessary to improve our guidelines in the use of the empirical antibiotic therapy.

  17. In vitro susceptibility test of sitafloxacin against resistant gram-negative bacilli isolated from Thai patients by disk diffusion method.

    Science.gov (United States)

    Thamlikitkul, Visanu; Tiengrim, Surapee

    2014-03-01

    To determine a correlation of minimum inhibitory concentration (MIC) of sitafloxacin determined by agar dilution method with inhibition zone diameter of sitafloxacin determined by disk diffusion method, and to determine inhibition zone, diameter breakpoints of sitafloxacin against resistant gram-negative bacilli isolated from Thai patients. The study bacteria were 332 clinical isolates of gram-negative bacilli including ESBL-producing E. coli, ESBL-producing K. pneumoniae, P. aeruginosa andA. baumannii. Each isolate of the present study bacteria was tested for minimum inhibitory concentration (MIC) of sitafloxacin by agar dilution method and inhibition zone diameter of sitafloxacin by disk diffusion method. The MICs and inhibition zone diameters of sitafloxacin against gram-negative bacilli were well correlated (correlation coefficient -0.926, p-value or = 15 mm had the least total error for determining susceptibility to sitafloxacin based on MIC value of sitafloxacin but the inhibition zone diameter > or = 16 mm had less false susceptibility than that of > or = 15 mm when compared with sitafloxacin MIC or = 19 mm had the least total error for determining susceptibility to sitafloxacin based on MIC value of sitafloxacin but the inhibition zone diameter > or = 18 mm had less false susceptibility than that of > or = 19 mm when compared with sitafloxacin MIC gram-negative bacilli by disk diffusion method, the inhibition zone diameter > or = 16 mm and > or = 18 mm seem to be the appropriate breakpoints for susceptibility for resistant gram-negative bacilli isolated from urine and blood, respectively, since the serum concentration of sitafloxacin is rather low whereas the urinary concentration of sitafloxacin is much higher.

  18. Evaluation of Methods for AmpC Beta-Lactamase in Gram Negative Clinical Isolates from Tertiary Care Hospitals

    OpenAIRE

    Singhal S; Mathur T; Khan S; Upadhyay D; Chugh S; Gaind R; Rattan A

    2005-01-01

    The purpose of this study was to simultaneously screen for Extended-spectrum b-lactamases (ESBL) and AmpC b-lactamases in gram negative clinical isolates from four tertiary care hospitals and further to compare two detection methods three-dimensional extraction method and AmpC disk test for AmpC b-lactamases. A total of 272 isolates were screened for ESBL and AmpC β-lactamase by modified double disk approximation method (MDDM). Synergy observed between disks of ceftazidime/cefotaxime a...

  19. In vitro activity of tigecycline and comparators against gram-negative bacteria isolated from a tertiary hospital in Alexandria, Egypt.

    Science.gov (United States)

    Mohamed, Nelly M; Youssef, Alaa A F

    2011-12-01

    The emergence of infections caused by multidrug-resistant Gram-negative bacteria, in particular Pseudomonas aeruginosa, Acinetobacter baumannii, and Klebsiella pneumoniae, has necessitated the search for alternative therapy by either introducing new agents or renewing interest in old agents. This study compares the in vitro activity of tigecycline (TIG), recently introduced to Egyptian market, to other potentially active antimicrobials as Colistin (COL), imipenem (IPM), levofloxacin (LEV), and piperacillin/tazobactam (PIP/TAZ) against 67 Gram-negative clinical isolates obtained from El- Meery Hospital in Alexandria, Egypt. El-Meery Hospital is a 1,500-bed tertiary teaching hospital where TIG has not been previously used. Based on MIC(90)s, TIG was found to be a comparator to IPM and COL (MIC(90)= 8 μg/ml). LEV and PIP/TAZ were less active than TIG exhibiting high MIC(90)s. TIG inhibited 100% of Escherichia coli and K. pneumoniae and 60% of Ps. aeruginosa and A. baumannii isolates. In time-kill studies against IPM-resistant isolates, TIG showed bactericidal activity after 6 hours of contact against the Enterobacteriaceae isolates and after 3 hours for the tested Ps. aeruginosa isolates at 4× and 8× MIC. Against A. baumannii, TIG exerted a bacteriostatic effect. TIG demonstrated variable ability to suppress biofilm formation affecting mainly E. coli and A. baumannii isolates. These results point TIG to be a promising agent in treatment of infections caused by strains for which adequate therapy has been limited. As far as we know, this is the first report evaluating the in vitro activity of TIG against Egyptian clinical isolates.

  20. Biofilm Matrix Composition Affects the Susceptibility of Food Associated Staphylococci to Cleaning and Disinfection Agents

    Science.gov (United States)

    Fagerlund, Annette; Langsrud, Solveig; Heir, Even; Mikkelsen, Maria I.; Møretrø, Trond

    2016-01-01

    Staphylococci are frequently isolated from food processing environments, and it has been speculated whether survival after cleaning and disinfection with benzalkonium chloride (BC)-containing disinfectants is due to biofilm formation, matrix composition, or BC efflux mechanisms. Out of 35 food associated staphylococci, eight produced biofilm in a microtiter plate assay and were identified as Staphylococcus capitis (2), S. cohnii, S. epidermidis, S. lentus (2), and S. saprophyticus (2). The eight biofilm producing strains were characterized using whole genome sequencing. Three of these strains contained the ica operon responsible for production of a polysaccharide matrix, and formed a biofilm which was detached upon exposure to the polysaccharide degrading enzyme Dispersin B, but not Proteinase K or trypsin. These strains were more tolerant to the lethal effect of BC both in suspension and biofilm than the remaining five biofilm producing strains. The five BC susceptible strains were characterized by lack of the ica operon, and their biofilms were detached by Proteinase K or trypsin, but not Dispersin B, indicating that proteins were major structural components of their biofilm matrix. Several novel cell wall anchored repeat domain proteins with domain structures similar to that of MSCRAMM adhesins were identified in the genomes of these strains, potentially representing novel mechanisms of ica-independent biofilm accumulation. Biofilms from all strains showed similar levels of detachment after exposure to alkaline chlorine, which is used for cleaning in the food industry. Strains with qac genes encoding BC efflux pumps could grow at higher concentrations of BC than strains without these genes, but no differences were observed at biocidal concentrations. In conclusion, the biofilm matrix of food associated staphylococci varies with respect to protein or polysaccharide nature, and this may affect the sensitivity toward a commonly used disinfectant. PMID:27375578

  1. Biofilm matrix composition affects the susceptibility of food associated staphylococci to cleaning and disinfection agents

    Directory of Open Access Journals (Sweden)

    Annette eFagerlund

    2016-06-01

    Full Text Available Staphylococci are frequently isolated from food processing environments, and it has been speculated whether survival after cleaning and disinfection with benzalkonium chloride-containing disinfectants is due to biofilm formation, matrix composition or benzalkonium chloride efflux mechanisms. Out of 35 food associated staphylococci, eight produced biofilm in a microtiter plate assay and were identified as Staphylococcus capitis (2, S. cohnii, S. epidermidis, S. lentus (2, and S. saprophyticus (2. The eight biofilm producing strains were characterized using whole genome sequencing. Three of these strains contained the ica operon responsible for production of a polysaccharide matrix, and formed a biofilm which was detached upon exposure to the polysaccharide degrading enzyme Dispersin B, but not Proteinase K or trypsin. These strains were more tolerant to the lethal effect of benzalkonium chloride both in suspension and biofilm than the remaining five biofilm producing strains. The five benzalkonium chloride susceptible strains were characterized by lack of the ica operon, and their biofilms were detached by Proteinase K or trypsin, but not Dispersin B, indicating that proteins were major structural components of their biofilm matrix. Several novel cell wall anchored repeat domain proteins with domain structures similar to that of MSCRAMM adhesins were identified in the genomes of these strains, potentially representing novel mechanisms of ica-independent biofilm accumulation. Biofilms from all strains showed similar levels of detachment after exposure to alkaline chlorine, which is used for cleaning in the food industry. Strains with qac genes encoding benzalkonium chloride efflux pumps could grow at higher concentrations of benzalkonium chloride than strains without these genes, but no differences were observed at biocidal concentrations. In conclusion, the biofilm matrix of food associated staphylococci varies with respect to protein or

  2. Isolation and molecular characterization of multidrug-resistant Gram-negative bacteria from imported flamingos in Japan

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    Fukumoto Yukio

    2009-11-01

    Full Text Available Abstract Imported animals, especially those from developing countries, may constitute a potential hazard to native animals and to public health. In this study, a new flock of lesser flamingos imported from Tanzania to Hiroshima Zoological Park were screened for multidrug-resistant Gram-negative bacteria, integrons and antimicrobial resistance genes. Thirty-seven Gram-negative bacterial isolates were obtained from the flamingos. Seven isolates (18.9% showed multidrug resistance phenotypes, the most common being against: ampicillin, streptomycin, tetracycline, trimethoprim/sulfamethoxazole and nalidixic acid. Molecular analyses identified class 1 and class 2 integrons, β-lactamase-encoding genes, blaTEM-1 and blaCTX-M-2 and the plasmid-mediated quinolone resistance genes, qnrS and qnrB. This study highlights the role of animal importation in the dissemination of multidrug-resistant bacteria, integrons and antimicrobial resistance genes from one country to another.

  3. Helicobacter ganmani sp nov., a urease-negative anaerobe isolated from the intestines of laboratory mice

    DEFF Research Database (Denmark)

    Robertson, B.R.; O'Rourke, J.L.; Vandamme, P.

    2001-01-01

    Spiral bacteria were isolated from the intestines of laboratory mice during a study examining the presence of Helicobacter species and other spiral organisms naturally infecting mice maintained at four different animal facilities in Sydney, Australia. One group of 17 isolates, cultured from mice...... from three of the four facilities, were found to be helicobacters but did not fall within any of the 18 currently recognized species. These isolates were unusual in that they only grew anaerobically at 37 degreesC and were incapable of growth under microaerobic conditions. Like Helicobacter rodentium...... and found to be identical to one another. H. rodentium was the most closely related species in terms of 16S rDNA sequence similarity (98.2%). Numerical analysis of whole-cell proteins by SDS-PAGE for nine isolates was carried out with a comparison to all known Helicobacter species, including newly...

  4. Negative visuospatial priming in isolation-reared rats: Evidence of resistance to the disruptive effects of amphetamine.

    Science.gov (United States)

    Amitai, Nurith; Powell, Susan; Weber, Martin; Swerdlow, Neal R; Young, Jared W

    2015-12-01

    Negative visuospatial priming (NP) represents a quantifiable measure of inhibitory information processing that is disrupted in several neurodevelopmental and psychiatric disorders, including schizophrenia. We developed a novel rodent NP task to investigate mechanisms underlying NP and its role in various disorders, and to test potential therapeutics. In the present studies, we further characterized this novel paradigm by investigating whether NP is disrupted in rats reared in isolation, a developmental manipulation that produces a range of abnormalities in behavior, neurochemistry, and brain structure that mirror aspects of schizophrenia pathology. We also further explored the role of monoaminergic signaling in NP and the effects of isolation rearing by challenging both socially reared and isolation-reared rats with D-amphetamine during the NP task. Although fewer isolation-reared animals learned the complex NP task, those that learned exhibited unaffected NP compared with socially reared rats. Consistent with previous reports, D-amphetamine impaired NP and increased motor impulsivity in socially reared rats. In contrast, D-amphetamine did not affect NP or motor impulsivity in isolation-reared rats. These data confirm a monoaminergic influence on NP behavior and indicate that rats reared in isolation have altered dopaminergic sensitivity.

  5. Diversity of mecA and ccrB in SCCmec type III of methicillin-resistant staphylococci.

    DEFF Research Database (Denmark)

    Damborg, Peter Panduro; Westh, Henrik; Boye, Kit;

    Background: Our research group recently showed that SCCmec type III is by far the most common type amongst methicillin-resistant coagulase-negative staphylococci (CoNS) of animal origin, in particular isolates within the Staphylococcus sciuri-group (S. sciuri, S. vitulinus, S. lentus and S....... pseudintermedius (n=2), S. vitulinus (n=2), S. sciuri (n=2), S. haemolyticus (n=1) and S. equorum (n=1) harbouring SCCmec III elements was subjected to partial sequence analysis of mecA and ccrB according to previously published protocols (Oliveira et al., JAC, 2006; Stephens et al., AAC, 2007). The sequences were...... positions. As for ccrB, species-specific clustering of mecA sequences was not observed as exemplified by mecA allele 5 being present in four species (S. pseudintermedius, S. sciuri, S. haemolyticus and S. vitulinus). Similarly, mecA allele 1 was found in three species (S. aureus, S. capitis and S...

  6. Isolation and characterization of a novel catalase-negative, urease-positive Campylobacter from cattle faeces

    DEFF Research Database (Denmark)

    Atabay, H.I.; Corry, J.E.L.; On, S.L.W.

    1997-01-01

    Forty-four strains of a phenotypically unique Campylobacter were isolated from the faeces of 26 of 45 cows in a single herd. Isolation involved enrichment and membrane filtration onto blood agar or plating onto cefoperazone amphotericin teicoplanin agar. The strains exhibited phenotypic character....... Their sensitivity to antibiotics normally used in selective media for campylobacters might explain why they have not previously been encountered. Their ecological significance and importance with respect to human and animal disease remain to be assessed....

  7. Antimicrobial resistance patterns among Gram-negative bacilli isolated from patients with nosocomial infections: Disk diffusion versus E-test

    Directory of Open Access Journals (Sweden)

    Hadadi A

    2007-08-01

    Full Text Available Background: The object of this study was to investigate the antimicrobial resistance pattern among common nosocomial Gram-negative bacilli isolated from patients with nosocomial infections. Methods: From June 2004 to December 2005, 380 isolates of common Gram-negative bacilli (Klebsiella, Pseudomonas, Acinetobacter and E. coli from 270 patients with nosocomial infections in Sina and Imam Hospitals, Tehran, Iran, were evaluated for susceptibility to Imipenem, Cefepime, Ciprofloxacine, Ceftriaxone and Ceftazidime by Disc diffusion and E-test methods. Results: The most frequent pathogens isolated were Klebsiella spp. (40%, followed by Pseudomonas (28%, Acinetobacter spp. (20% and E. coli (12%. The most active antibiotic was imipenem (84%. 26% of all isolates were sensitive to Cefepime, 26% to Ciprofloxacin, 20% to Ceftazidime and 10% to Ceftrixone. The susceptibility rates of Klebsiella to Imipenem, cefepime, ciprofloxacin, Ceftazidime and Ceftriaxone were 91, 25, 21, 13 and 7 percent, respectively and 91, 19, 17, 21 and 21 percent, respectively, for E. coli. Among Acineto- bacter spp., the susceptibility rate was 77% for Imipenem and 21% for Ciprofloxacin. Among Pseudomonas spp., 75% of isolates were susceptible to Imipenem and 39% to Ciprofloxacin. The comparison of the resistance status of microorganisms by both Disc diffusion and E-test methods showed a clinically noticeable agreement between these two tests. Conclusions: Since antibiotic resistance among Gram-negative bacilli has increased, enforcement of policy regarding proper antibiotic use is urgently needed in order to delay the development of resistance. Although it is widely accepted that E-test is more accurate in determining the resistance of microorganisms, our study showed that the Disc diffusion test will give the same results in most occasions and is therefore still considered useful in clinical practice.

  8. Screening of Extended Spectrum Beta lactamase Producing Gram Negative Bacilli Isolated from Clinical Cases

    OpenAIRE

    H Hoseinzadegan; A Hassani; M Azadpoor; S Soleimannezhad; F. Mohamadi

    2007-01-01

    Background and objectives:(ESBL) strain is one of the emerging health related problems in the world recently.Some of the species of the gram-negative bacilli including Klebsiella Pneumonia &Escherichia Coli are well known ESBL producing among bacteria, and they causeuncontrollable infections. This Cross-sectional study was designed to asses theESBL producing gram negative bacilli among inpatients of Shohada-ye- ashayerhospital (Khorram Abad).Extended Spectrum Betalactamase producingMaterials ...

  9. SELECTIVE ISOLATION OF A GRAM NEGATIVE CARBAMATE PESTICIDE DEGRADING BACTERIUM FROM BRINJAL CULTIVATED SOIL

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    Dilara Islam Sharif

    2013-01-01

    Full Text Available A bacterial species having the ability to grow in the presence of carbosulfan pesticide “Marshall” was isolated from Solanum melongena (brinjal cultivated soil in Khulna region, Bangladeh, having a history of pesticide usage. The strain was morphologically and biochemically identified to belong to the genera Pseudomonas. A comparative study of growth of this strain with other isolated bacteria showed its ability to grow in the presence of different concentrations of Marshall. The susceptibility of the stain to Marshall was also assessed through disk diffusion assay which showed the strain to be resistant at concentrations of Marshal commonly used under field conditions. The selected strain also showed its capability to degrade Marshall through observed characteristics on sublimated agar plates. The biodegradation capability of the strain isolated in this study can be valuable for further study towards bioremediation of pesticide contaminated soils.

  10. Emergence of integron borne PER-1 mediated extended spectrum cephalosporin resistance among nosocomial isolates of Gram-negative bacilli

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    Anand Prakash Maurya

    2015-01-01

    Full Text Available Background & objectives: Pseudomonas extended resistant (PER enzymes are rare type of extended-spectrum beta lactamases (ESBLs that confer third generation cephalosporin resistance. These are often integron borne and laterally transmitted. The aim of the present study was to investigate the emergence of integron borne cephalosporin resistant PER-1 gene in diverse incompatibility (Inc group plasmids among Gram-negative bacteria. Methods: A total of 613 consecutive, non-duplicate, Gram-negative bacteria of Enterobacteriaceae family and non-fermenting Gram-negative bacteria were isolated from different clinical specimens during a period of 18 months. For amplification and detection of blaPER, multiplex PCR was done. For understanding the genetic environment of blaPER-1, integrase gene PCR and cassette PCR (59 be was performed. Gene transferability experiment was carried out and PCR based replicon typing was performed for incompatibility group typing of plasmids using 18 pairs of primers. An inhibitor based method was used for phenotypic detection of intrinsic resistance. Results: Multiplex PCR and sequencing confirmed that 45 isolates were harbouring blaPER-1. Both class 1 and class 2 integrons were observed among them. Integrase and cassette PCR (59 be PCR results confirmed that the resistant determinant was located within class 1 integron. Transformation and conjugation experiments revealed that PER-1 was laterally transferable and disseminated through diverse Inc plasmid type. Efflux pump mediated carbapenem resistance was observed in all isolates. All isolates belonged to heterogenous groups. Interpretation & conclusions: This study demonstrates the dissemination of cephalosporins resistant, integron borne blaPER-1 in hospital setting in this part of the country and emphasizes on the rational use of third generation cephalosporins to slow down the expansion of this rare type of ESBL gene.

  11. Kaposi′s sarcoma: HIV-negative man with isolated penile localization

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    Soufiane Mellas

    2010-07-01

    Full Text Available Kaposi′s sarcoma is the malignant proliferation of the endothelial cell vessels. Its genesis is still unclear; however, it seems to be related to the herpes virus infection (HHV-8. This neoplasia usually affects the lower limbs and the affected persons are mostly from the Mediterranean region. The exclusive penile localization of the Kaposi′s sarcoma in a patient with a negative HIV serologia is exceptional. Our case is of a 73-year old patient with a negative HIV serology presenting an exclusive penile localization of the Kaposi′s sarcoma treated by radiotherapy.

  12. Isolation of Bartonella henselae from a serologically negative cat in Bloemfontein, South Africa

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    A-M Pretorius

    1999-07-01

    Full Text Available Sera collected from apparently healthy 6-12-month-old cats (n = 31 presented to the Society for the Prevention of Cruelty to Animals Veterinary Clinic in Bloemfontein for neutering were tested for antibodies reactive to Bartonella henselae (Houston-1 strain by indirect fluorescent antibody testing. Whole blood collected from the cats was used in isolation experiments and subsequent identification of Bartonella species was based on comparison of the nucleotide base sequence of polymerase chain reaction-amplified citrate synthase gene fragments. While none of the cats had antibodies reactive with B. henselae at titres > 1/64, an organism with a partial citrate synthase gene sequence identical to that of B. henselae (Houston-1 was isolated from 1 cat.

  13. Isolation of Extended Spectrum β-Lactamase Producing Gram Negative Bacilli from Wound Swab with Their Antimicrobial Susceptibility Pattern.

    Science.gov (United States)

    Islam, T A; Shamsuzzaman, S M

    2016-10-01

    Extended Spectrum β-lactamases (ESBLs) producing organisms have become the major clinical concern worldwide. The present study was undertaken to see the frequency of ESBLs producing gram-negative bacilli with their antibiogram in post surgical wound swab collected over a period of 12 months from July 2011 to June 2012 at Dhaka Medical College Hospital. Among 200 samples 121(60.5%) gram negative bacilli and 52(26%) gram-positive bacteria were isolated. Escherichia coli (36.42%) was the most predominant gram-negative bacilli followed by Klebsiella species (9.83%) and Pseudomonas aeruginosa (8.67%). Thirty four (28.1%) isolates were detected as ESBLs producers by double-disc synergy test (DDST) and the prevalence among Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa were 33.33%, 35.29% and 26.67% respectively. All the ESBL producing strains were sensitive to imipenem but they were significantly more resistant to ciprofloxacin, doxycycline, amoxiclav, co-trimoxazole, azithromycin and gentamycin than non-ESBLs producers (p<0.01). The finding suggests more effective strategies are needed to control the spread of these resistant organisms.

  14. Metallo- β-lactamases among Multidrug Resistant (MDR Gram Negative Bacteria Isolated from Clinical Specimens during 2009 in Sanandaj, Kurdistan Province

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    Himen Salimizand

    2012-08-01

    Full Text Available Background: Today, there are numerous reports about emerging multi drug resistant gram negative bacteria all around the world, especially in ICUs. Rarely, Metallo-β-lactamase (MBL enzymes are responsible for these cases. Study of MBLs for diagnosing and preventing distribution of the origin of infection are critical issues. In addition, we would like to compare the efficacy of Iranian and foreign- made antibiotic disks. Materials and Methods: During 2009 all entered clinical specimens to the laboratory tested for detecting gram negative bacteria. Isolated bacteria were tested by Kirby-Bauer method to antibiotic susceptibility test by Iranian and foreign (MAST disks. For gram negative carbapenem resistant isolates, PCR technique used to detect VIM, GIM, and SIM variants of MBLs.Results: During one year, 17890 clinical specimens referred Besat laboratory. The most specimen was Urine (8172 followed by blood culture (5190 that in which 1110 gram negative and positives isolated. Out of which, 778 (70% of isolates were gram negatives. MDR gram negatives were 157 (20.2%. Imipenem and meropenem were the most efficient antibiotics (all susceptible and ceftriaxone was the least (19 % susceptible. E. coli was the most prevalent isolate. 79 Gram negative isolates (10.1% were resistant to Iranian-made discs but all susceptible for foreign ones. All 79 isolates were tested by PCR for MBL genes, that, all were negative. Besides, Iranian imipenem and cefepime disks have had distinguishable difference in susceptibility of isolates.Conclusion: Fortunately, none of gram negative isolates were MBL producer, which revealed no colonization of MBL producing bacteria. Iranian-made disks appear efficient except for imipenem and cefepime.

  15. Biodegradation of geosmin by a novel Gram-negative bacterium; isolation, phylogenetic characterisation and degradation rate determination.

    Science.gov (United States)

    Hoefel, Daniel; Ho, Lionel; Monis, Paul T; Newcombe, Gayle; Saint, Christopher P

    2009-06-01

    Biologically active sand filters within water treatment plants (WTPs) are now recognised as an effective barrier for the removal of geosmin. However, little is known regarding the actual microbiological processes occurring or the bacteria capable of degrading geosmin. This study reports the enrichment and isolation of a Gram-negative bacterium, Geo48, from the biofilm of a WTP sand filter where the isolate was shown to effectively degrade geosmin individually. Experiments revealed that Geo48 degraded geosmin in a planktonic state by a pseudo-first-order mechanism. Initial geosmin concentrations ranging from 100 to 1000ng/l were shown to directly influence geosmin degradation in reservoir water by Geo48, with rate constants increasing from 0.010h(-1) (R(2)=0.93) to 0.029h(-1) (R(2)=0.97) respectively. Water temperature also influenced degradation of geosmin by Geo48 where temperatures of 11, 22 and 30 degrees C resulted in rate constants of 0.017h(-1) (R(2)=0.98), 0.023h(-1) (R(2)=0.91) and 0.019h(-1) (R(2)=0.85) respectively. Phylogenetic analysis using the 16S rRNA gene of Geo48 revealed it was a member of the Alphaproteobacteria and clustered with 99% bootstrap support with an isolate designated Geo24, a Sphingopyxis sp. previously described as degrading geosmin but only as a member of a bacterial consortium. Of the previously described bacteria, Geo48 was most similar to Sphingopyxis alaskensis (97.2% sequence similarity to a 1454bp fragment of the 16S rRNA gene). To date, this is the only study to report the isolation and characterisation of a Gram-negative bacterium from a biologically active sand filter capable of the sole degradation of geosmin.

  16. Multiplex real-time PCR assay for rapid detection of methicillin-resistant staphylococci directly from positive blood cultures.

    Science.gov (United States)

    Wang, Hye-Young; Kim, Sunghyun; Kim, Jungho; Park, Soon-Deok; Uh, Young; Lee, Hyeyoung

    2014-06-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is the most prevalent cause of bloodstream infections (BSIs) and is recognized as a major nosocomial pathogen. This study aimed to evaluate a newly designed multiplex real-time PCR assay capable of the simultaneous detection of mecA, S. aureus, and coagulase-negative staphylococci (CoNS) in blood culture specimens. The Real-MRSA and Real-MRCoNS multiplex real-time PCR assays (M&D, Republic of Korea) use the TaqMan probes 16S rRNA for Staphylococcus spp., the nuc gene for S. aureus, and the mecA gene for methicillin resistance. The detection limit of the multiplex real-time PCR assay was 10(3) CFU/ml per PCR for each gene target. The multiplex real-time PCR assay was evaluated using 118 clinical isolates from various specimen types and a total of 350 positive blood cultures from a continuous monitoring blood culture system. The results obtained with the multiplex real-time PCR assay for the three targets were in agreement with those of conventional identification and susceptibility testing methods except for one organism. Of 350 positive bottle cultures, the sensitivities of the multiplex real-time PCR kit were 100% (166/166 cultures), 97.2% (35/36 cultures), and 99.2% (117/118 cultures) for the 16S rRNA, nuc, and mecA genes, respectively, and the specificities for all three targets were 100%. The Real-MRSA and Real-MRCoNS multiplex real-time PCR assays are very useful for the rapid accurate diagnosis of staphylococcal BSIs. In addition, the Real-MRSA and Real-MRCoNS multiplex real-time PCR assays could have an important impact on the choice of appropriate antimicrobial therapy, based on detection of the mecA gene.

  17. Impact of Antimicrobial Stewardship Programme on Carbapenem Resistance in Gram Negative Isolates in an Indian Tertiary Care Hospital

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    Namita Jaggi

    2012-01-01

    Full Text Available Problem statement: Increasing Antimicrobial resistance in the World is constantly becoming a Global threat and there is an urgent need to prevent its spread. Various studies of last decade have shown reduced trends of antimicrobial resistance in the pathogens as an outcome of the Antimicrobial Stewardship Programs. In view of this, the present four years’ study was carried out to analyse the impact of Antimicrobial Stewardship Programs on carbapenem resistance in Gram negative isolates in a Tertiary care hospital in India. It involved a retrospective analysis of carbapenem resistance in Gram negatives for one year (July 2007 to June 2008, followed by prospective evaluation of the impact of stewardship interventions on resistance patterns (July 2008 to Jun 2011. Approach: Our study was staged into four parts: (1 July 2007 to June 2008: Resistance patterns of Gram negative isolates-E.coli, Klebsiella, Pseudomonas and Acinetobacter baumannii towards carbapenems were studied. (2 July 2008: Phase I intervention programme Implementation of an antibiotic policy in the hospital. (3 July 2008 to June 2010: The Impact of Phase I intervention programme was assessed subsequently. (4 July 2010 to June 2011: Phase II intervention programme: Formation and effective functioning of the antimicrobial stewardship committee. Results: The percentage resistance towards carbapenems in E.coli, Klebsiella, Pseudomonas and A. baumannii from July 2007-June 2008 was 1.07, 13.1, 21.3 and 12.5% respectively. Phase I intervention programme was initiated in July 2008 and Phase II in July 2010 and a subsequent reduction of 4.03% was observed in the carbapenem resistant Pseudomonas in the last stage of study period following the interventions. However the resistance in the other Gram negatives (E. coli, Klebsiella and A. baumannii rose and then stabilized. Conclusion: An antimicrobial stewardship programme with sustained and multifaceted efforts is essential to control the

  18. Antimicrobial susceptibility testing of newer quinolones against gram positive and gram negative clinical isolates.

    Science.gov (United States)

    Iffat, Wajiha; Shoaib, Muhammad Harris; Muhammad, Iyad Naeem; Rehana; Tasleem, Samiah; Gauhar, Shahnaz

    2010-07-01

    Antibiotic resistance development is an ongoing process associated with irrational antibiotic use. WHO recommends regular surveillance programs for monitoring of antibiotic resistance. The present study is a step in this direction. A total of 124 clinical isolates of Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae and Pseudomonas aeruginosa were collected from different hospitals in Karachi. In vitro antimicrobial susceptibility studies were carried out by agar dilution method using newer quinolones that included Gatifloxacin and Levofloxacin. It was observed that 50% (n=30) isolates of Staphylococcus aureus were resistant to gatifloxacin. Gatifloxacin was more active against Pseudomonas aeruginosa (n=23) and showing complete susceptibility with MIC 1mg/L except for three very resistant strains that shown resistance at even higher concentrations. Escherichia coli (n=45) has shown 15.5% and Klebsiella pneumoniae (n=26) 34.61% resistance to gatifloxacin. Levofloxacin was more active against Staphylococcus aureus and Escherichia coli showing complete susceptibility at 0.5 mg /L concentration. Pseudomonas aeruginosa and Klebsiella pneumoniae were found to be resistant to Levofloxacin showing 36.36% and 23.08% resistance respectively. The study strongly recommends the adherence to the antibiotic policy and regular susceptibility testing to overcome the problem associated with antimicrobial resistance.

  19. Phenotypic methods for detection of various β-lactamases in Gram-negative clinical isolates: Need of the hour

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    Neena V Nagdeo

    2012-01-01

    Full Text Available Background: Many clinical laboratories have problems detecting various β-lactamases. Confusion exists about the importance of these resistance mechanisms, optimal test methods, and appropriate reporting conventions. It is more imperative to use various phenotypic methods for detection of various β-lactamases in routine microbiology laboratory on day-to-day basis to prevent antimicrobial resistance by evidence-based judicious use of antimicrobials. Aims: In view of the multidrug-resistant organisms being reported world over, we planned a cross-sectional prospective analytical study to determine resistance mechanism by various β-lactamases in Gram-negative clinical isolates using various phenotypic methods. Materials and Methods: All nonrepeat, nonenteric clinical isolates of Gram-negative bacilli, resistant to at least two third-generation cephalosporins, were first screened by Novel disc placement method, and isolates showing multiple mechanisms of resistance and reduced zone of inhibition for imipenem were further confirmed for AmpC and metallo β-lactamases. Statistical Analysis: All the data was managed and analyzed in Microsoft Excel. Results: Out of 807 isolates tested, as many as 795 (98.51% revealed the presence of extended-spectrum β-lactamases (ESBLs. Only 10 isolates of Escherichia coli and 2 of Klebsiella pneumoniae did not show production of ESBL. A total of 450 (55.76% isolates produced single enzyme,while 345 (42.75% strains revealed multiple enzyme production simultaneously. Only ESBL production was seen in 315 (39.03% strains, only AmpC in 75 (9.29% and only MBL in 60 (7.44% strains, while ESBL and AmpC together were seen in 219 (27.14% and AmpC plus MBL in 92 (11.40% strains. However, ESBL plus MBL were never observed together. All three enzymes were simultaneously detected in 34 (4.21% strains. Conclusion: This innovative method of disc placement makes it easy, affordable, and reliable method for routine use by basic

  20. Investigated of ampC in Carbapenem Resistant Gram-Negative Bacteria Isolated from Burned Patients

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    Leila Azimi

    2015-10-01

    Full Text Available Background: Gram-Negative bacteria are the most cause of nosocomial infection especially in burned patients. Carbapenem resistant strains can limit seriously the choice of antibiotic therapy. AmpC can make resistance to carbapenem and detection of that can be useful for identification ofcarbapenem resistant. The aim of this study was identification of ampC in most prevalent cause of nosocomial infection.Methods: boronic acid combined with meropenem in combination disc method was used for phenotypic method and PCR was used for molecular identification of ampC.Results: Fifty one strains showed positive results in phenotypic method but 119 strains were harbored ampC gene. Combination disc method with meropenem and boronic acid showed 34.4% sensitivity and 78.5% specificity according to the results of this study.Conclusions: the results of this study were indicated the more prevalent of ampC in carbapenem resistant Gram-Negative bacteria. On the other hand use of boronic acid combined with meropenem showed low sensitivity for detection of ampC.

  1. Pharmacodynamic Profiling of Antimicrobials against Gram-negative Respiratory Isolates from Canadian Hospitals

    Directory of Open Access Journals (Sweden)

    Rebecca A. Keel

    2011-01-01

    Full Text Available The objective of this study was to assess the profile of a variety of dosing regimens for common intravenous antibiotics against contemporary Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa isolates collected in Canada during 2009, using pharmacodynamic modelling techniques. Monte Carlo simulation was conducted for standard and/or prolonged infusion regimens of cefepime, ceftazidime, ceftriaxone, ciprofloxacin, doripenem, ertapenem, meropenem and piperacillin/tazobactam. The cumulative fraction of response (CFR was calculated using bactericidal targets for each regimen against each species. All cefepime, doripenem, ertapenem and meropenem regimens achieved optimal exposures against Enterobacteriaceae, whereas target attainment was organism and dose dependent for the other agents. These results support that the currently recommended antimicrobial dosing regimens generally attain acceptable exposures to achieve the requisite pharmacodynamic targets against the Enterobacteriaceae species; however, they fall short of obtaining optimal bactericidal exposures against P aeruginosa.

  2. Ice-nucleation negative fluorescent pseudomonads isolated from Hebridean cloud and rain water produce biosurfactants

    Directory of Open Access Journals (Sweden)

    H. E. Ahern

    2006-10-01

    Full Text Available Microorganisms were discovered in clouds over 100 years ago but information on bacterial community structure and function is limited. Clouds may not only be a niche within which bacteria could thrive but they might also influence dynamic processes using ice nucleating and cloud condensing abilities. Cloud and rain samples were collected from two mountains in the Outer Hebrides, NW Scotland, UK. Community composition was determined using a combination of amplified 16S ribosomal DNA restriction analysis and sequencing. 256 clones yielded 100 operational taxonomic units (OTUs of which half were related to bacteria from terrestrial psychrophilic environments. Cloud samples were dominated by a mixture of fluorescent Pseudomonas spp., some of which have been reported to be ice nucleators. It was therefore possible that these bacteria were using the ice nucleation (IN gene to trigger the Bergeron-Findeisen process of raindrop formation as a mechanism for dispersal. In this study the IN gene was not detected in any of the isolates using both polymerase chain reaction (PCR and differential scanning calorimetry (DSC. Instead 55% of the total isolates from both cloud and rain samples displayed significant biosurfactant activity when analyzed using the drop-collapse technique. All were characterised as fluorescent pseudomonads. Surfactants have been found to be very important in lowering atmospheric critical supersaturations required for the activation of aerosols into cloud condensation nuclei (CCN. It is also known that surfactants influence cloud droplet size and increase cloud lifetime and albedo. Some bacteria are known to act as CCN and so it is conceivable that these fluorescent pseudomonads are using surfactants to facilitate their activation from aerosols into CCN. This would allow water scavenging, countering desiccation, and assist in their widespread dispersal.

  3. Ice-nucleation negative fluorescent pseudomonads isolated from Hebridean cloud and rain water produce biosurfactants

    Science.gov (United States)

    Ahern, H. E.; Walsh, K. A.; Hill, T. C. J.; Moffett, B. F.

    2006-10-01

    Microorganisms were discovered in clouds over 100 years ago but information on bacterial community structure and function is limited. Clouds may not only be a niche within which bacteria could thrive but they might also influence dynamic processes using ice nucleating and cloud condensing abilities. Cloud and rain samples were collected from two mountains in the Outer Hebrides, NW Scotland, UK. Community composition was determined using a combination of amplified 16S ribosomal DNA restriction analysis and sequencing. 256 clones yielded 100 operational taxonomic units (OTUs) of which half were related to bacteria from terrestrial psychrophilic environments. Cloud samples were dominated by a mixture of fluorescent Pseudomonas spp., some of which have been reported to be ice nucleators. It was therefore possible that these bacteria were using the ice nucleation (IN) gene to trigger the Bergeron-Findeisen process of raindrop formation as a mechanism for dispersal. In this study the IN gene was not detected in any of the isolates using both polymerase chain reaction (PCR) and differential scanning calorimetry (DSC). Instead 55% of the total isolates from both cloud and rain samples displayed significant biosurfactant activity when analyzed using the drop-collapse technique. All were characterised as fluorescent pseudomonads. Surfactants have been found to be very important in lowering atmospheric critical supersaturations required for the activation of aerosols into cloud condensation nuclei (CCN). It is also known that surfactants influence cloud droplet size and increase cloud lifetime and albedo. Some bacteria are known to act as CCN and so it is conceivable that these fluorescent pseudomonads are using surfactants to facilitate their activation from aerosols into CCN. This would allow water scavenging, countering desiccation, and assist in their widespread dispersal.

  4. Non dominant-negative KCNJ2 gene mutations leading to Andersen-Tawil syndrome with an isolated cardiac phenotype.

    Science.gov (United States)

    Limberg, Maren M; Zumhagen, Sven; Netter, Michael F; Coffey, Alison J; Grace, Andrew; Rogers, Jane; Böckelmann, Doris; Rinné, Susanne; Stallmeyer, Birgit; Decher, Niels; Schulze-Bahr, Eric

    2013-05-01

    Andersen-Tawil syndrome (ATS) is characterized by dysmorphic features, periodic paralyses and abnormal ventricular repolarization. After genotyping a large set of patients with congenital long-QT syndrome, we identified two novel, heterozygous KCNJ2 mutations (p.N318S, p.W322C) located in the C-terminus of the Kir2.1 subunit. These mutations have a different localization than classical ATS mutations which are mostly located at a potential interaction face with the slide helix or at the interface between the C-termini. Mutation carriers were without the key features of ATS, causing an isolated cardiac phenotype. While the N318S mutants regularly reached the plasma membrane, W322C mutants primarily resided in late endosomes. Co-expression of N318S or W322C with wild-type Kir2.1 reduced current amplitudes only by 20-25 %. This mild loss-of-function for the heteromeric channels resulted from defective channel trafficking (W322C) or gating (N318S). Strikingly, and in contrast to the majority of ATS mutations, neither mutant caused a dominant-negative suppression of wild-type Kir2.1, Kir2.2 and Kir2.3 currents. Thus, a mild reduction of native Kir2.x currents by non dominant-negative mutants may cause ATS with an isolated cardiac phenotype.

  5. Prevalence and risk factors for colonization with methicillin resistant Staphylococcus aureus and other Staphylococci species in hospitalized and farm horses in Israel.

    Science.gov (United States)

    Tirosh-Levy, Sharon; Steinman, Amir; Carmeli, Yehuda; Klement, Eyal; Navon-Venezia, Shiri

    2015-11-01

    Methicillin-resistant staphylococci (MRS), and specifically Methicillin-resistant Staphylococcus aureus (MRSA) colonization or infection have become a serious emerging condition in equine hospitals, with complex concerns regarding animals, personnel and public health. The objectives of this study were to evaluate the prevalence and risk factors for colonization by Staphylococci, MRS, and MRSA among horses in Israel. Nasal swabs were collected from horses at 17 riding stables (n=206), and from hospitalized horses admitted to a veterinary hospital (n=84). Species identification was performed by pta gene PCR, RFLP analysis and sequencing. MRS was identified by the presence of mecA. Genetic relatedness of MRSA isolates was determined by spa typing and MLST. SCCmec-type and pvl gene were determined. Univariable and multivariable statistical analysis were used to identify potential risk factors. Colonization with Staphylococci was found among 3.8% of farm horses and 50.6% of hospitalized horses (pantibiotic use. This is the first surveillance study of Staphylococci in horses in Israel, and the first report on the presence of a unique MRSA strain among hospital horses, recognizing the veterinary hospital as a potential reservoir for MRSA, an antibiotic resistant pathogen with human relevance.

  6. Characterisation of polyacetylenes isolated from carrot (Daucus carota) extracts by negative ion tandem mass spectrometry.

    Science.gov (United States)

    Rai, Dilip K; Brunton, Nigel P; Koidis, Anastasios; Rawson, Ashish; McLoughlin, Padraig; Griffiths, William J

    2011-08-15

    The potential use of negative electrospray ionisation mass spectrometry (ESI-MS) in the characterisation of the three polyacetylenes common in carrots (Daucus carota) has been assessed. The MS scans have demonstrated that the polyacetylenes undergo a modest degree of in-source decomposition in the negative ionisation mode while the positive ionisation mode has shown predominantly sodiated ions and no [M+H](+) ions. Tandem mass spectrometric (MS/MS) studies have shown that the polyacetylenes follow two distinct fragmentation pathways: one that involves cleavage of the C3-C4 bond and the other with cleavage of the C7-C8 bond. The cleavage of the C7-C8 bond generated product ions m/z 105.0 for falcarinol, m/z 105/107.0 for falcarindiol, m/z 147.0/149.1 for falcarindiol-3-acetate. In addition to these product ions, the transitions m/z 243.2 → 187.1 (falcarinol), m/z 259.2 → 203.1 (falcarindiol), m/z 301.2 → 255.2/203.1 (falcarindiol-3-acetate), mostly from the C3-C4 bond cleavage, can form the basis of multiple reaction monitoring (MRM)-quantitative methods which are poorly represented in the literature. The 'MS(3) ' experimental data confirmed a less pronounced homolytic cleavage site between the C11-C12 bond in the falcarinol-type polacetylenes. The optimised liquid chromatography (LC)/MS conditions have achieved a baseline chromatographic separation of the three polyacetylenes investigated within 40 min total run-time.

  7. Characterization of Staphylococcus xylosus isolated from broiler chicken barn bioaerosol.

    Science.gov (United States)

    Vela, Javier; Hildebrandt, Kelsie; Metcalfe, Andrew; Rempel, Heidi; Bittman, Shabtai; Topp, Edward; Diarra, Moussa

    2012-12-01

    In this study we isolated and characterized Staphylococcus xylosus, a coagulase-negative staphylococcal species considered as commensal and one of the prevalent staphylococcal species found in poultry bioaerosol. Isolates were obtained using air samplers and selective phenylethyl alcohol agar for gram-positive bacteria during 35-d periods at different times of the day. A total of 200 colonies were recovered and after basic biochemical tests were performed, presumptive staphylococci were subsequently identified by API Staph strips. A total of 153 (76.5%) staphylococci were found, among which 84 were S. xylosus (46 and 38 isolated inside and outside, respectively). Biofilm formation was observed in 86.9% of S. xylosus isolates, whereas 79.8% of them showed hemolytic activity. There was a strong correlation (92.5%) between biofilm formation and hemolytic activity. All 84 S. xylosus isolates were susceptible to amikacin, ampicillin/sulbactam, chloramphenicol, ciprofloxacin, gentamycin, kanamycin, linezolid, trimethoprim/sulfamethoxazole, and vancomycin. Resistance to nalidixic acid (86.9%), novobiocin (85.7%), penicillin (70.2%), lincomycin (46.4%), oxacillin (42.9%), ampicillin (27.4%), tetracycline (21.4%), erythromycin (11.9%), bacitracin (10.7%), and streptomycin (2.4%) was observed among the isolates. Resistance to tetracycline, lincomycin, erythromycin, and β-lactam antibiotics was occasionally linked to the tetK, linA, ermB, and blaZ genes, respectively. Random amplification of polymorphic DNA results showed similarity of 15 to 99% between isolates collected outside and inside the barn, indicating genetic diversity of these isolates. Our study indicates that characterization of poultry bioaerosol coagulase-negative staphylococcal species such as S. xylosus is necessary for assessing their safety status for both poultry and humans.

  8. Emergence of Carbapenem Resistant Non-Fermenting Gram-Negative Bacilli Isolated in an ICU of a Tertiary Care Hospital.

    Science.gov (United States)

    Agarwal, Sonika; Kakati, Barnali; Khanduri, Sushant; Gupta, Shalini

    2017-01-01

    The emergence and spread of Multi-Drug Resistant (MDR) Non-Fermenting Gram-Negative Bacilli (NFGNB) in Intensive Care Units (ICU) and their genetic potential to transmit diverse antibiotic resistance regardless of their ability to ferment glucose poses a major threat in hospitals. The complex interplay of clonal spread, persistence, transmission of resistance elements and cell-cell interaction leads to the difficulty in controlling infections caused by these multi drug-resistant strains. Among non-fermenting Gram-negative rods, the most clinically significant species Pseudomonas aeruginosa, Acinetobacter baumannii and Stenotrophomonas maltophilia are increasingly acquiring resistant to carbapenems. Carbapenems once considered as a backbone of treatment of life threatening infections appears to be broken as the resistance to carbapenems is on rise. To document the prevalence of carbapenem resistance in non-fermenting Gram-negative bacilli isolated from patients with respiratory tract infections in the ICU of Himalayan Institute of Medical Sciences, Dehradun. This is a cross-sectional study conducted in ICU patients between October 2015 to March 2016. A total of 366 lower respiratory tract samples were collected from 356 patients with clinical evidence of lower respiratory tract infections in form of Endotracheal (ET) aspirate, Tracheal Tube (TT) aspirate and Bronchoalveolar Lavage (BAL) specimen. Organism identification and the susceptibility testing was done by using an automated system VITEK 2. Out of 366 samples received 99 NFGNB were isolated and most common sample was ET aspirate sample 256 (64.5%). Acinetobacter baumannii was the most common NFGNB isolated 63 (63.63%) followed by Pseudomonas aeruginosa 25 (25.25%), Elizabethkingia meningoseptica seven (7.07%) and Strenotrophomonas maltophilia four (4.04%). We observed that 90.5% Acinetobacter baumannii were resistant to imipenem and 95.2% resistant to meropenem, Pseudomonas aeruginosa came out to be 52

  9. Antibiotic Resistance of Gram Negatives isolates from loggerhead sea turtles (Caretta caretta) in the central Mediterranean Sea.

    Science.gov (United States)

    Foti, M; Giacopello, C; Bottari, Teresa; Fisichella, V; Rinaldo, D; Mammina, C

    2009-09-01

    Previous studies on fish and marine mammals support the hypothesis that marine species harbor antibiotic resistance and therefore may serve as reservoirs for antibiotic-resistance genetic determinants. The aim of this study was to assess the resistance to antimicrobial agents of Gram negative strains isolated from loggerhead sea turtles (Carettacaretta). Oral and cloacal swabs from 19 live-stranded loggerhead sea turtles, with hooks fixed into the gut, were analyzed. The antimicrobial resistance of the isolates to 31 antibiotics was assessed using the disk-diffusion method. Conventional biochemical tests identified Citrobacter spp., Proteus spp., Enterobacter spp., Escherichia spp., Providencia spp., Morganella spp., Pantoea spp., Pseudomonas spp. and Shewanella spp. Highest prevalences of resistance was detected to carbenicillin (100%), cephalothin (92.6%), oxytetracycline (81.3%) and amoxicillin (77.8%). The isolates showing resistance to the widest range of antibiotics were identified as Citrobacterfreundii, Proteusvulgaris, Providenciarettgeri and Pseudomonasaeruginosa. In this study, antibiotic resistant bacteria reflect marine contamination by polluted effluents and C.caretta is considered a bioindicator which can be used as a monitor for pollution.

  10. Multidrug Resistance in Quinolone-Resistant Gram-Negative Bacteria Isolated from Hospital Effluent and the Municipal Wastewater Treatment Plant.

    Science.gov (United States)

    Vaz-Moreira, Ivone; Varela, Ana Rita; Pereira, Thamiris V; Fochat, Romário C; Manaia, Célia M

    2016-03-01

    This study is aimed to assess if hospital effluents represent an important supplier of multidrug-resistant (MDR) Gram-negative bacteria that, being discharged in the municipal collector, may be disseminated in the environment and bypassed in water quality control systems. From a set of 101 non-Escherichia coli Gram-negative bacteria with reduced susceptibility to quinolones, was selected a group of isolates comprised by those with the highest indices of MDR (defined as nonsusceptibility to at least one agent in six or more antimicrobial categories, MDR ≥6) or resistance to meropenem or ceftazidime (n = 25). The isolates were identified and characterized for antibiotic resistance phenotype, plasmid-mediated quinolone resistance (PMQR) genes, and other genetic elements and conjugative capacity. The isolates with highest MDR indices were mainly from hospital effluent and comprised ubiquitous bacterial groups of the class Gammaproteobacteria, of the genera Aeromonas, Acinetobacter, Citrobacter, Enterobacter, Klebsiella, and Pseudomonas, and of the class Flavobacteriia, of the genera Chryseobacterium and Myroides. In this group of 25 strains, 19 identified as Gammaproteobacteria harbored at least one PMQR gene (aac(6')-Ib-cr, qnrB, qnrS, or oqxAB) or a class 1 integron gene cassette encoding aminoglycoside, sulfonamide, or carbapenem resistance. Most of the E. coli J53 transconjugants with acquired antibiotic resistance resulted from conjugation with Enterobacteriaceae. These transconjugants demonstrated acquired resistance to a maximum of five classes of antibiotics, one or more PMQR genes and/or a class 1 integron gene cassette. This study shows that ubiquitous bacteria, other than those monitored in water quality controls, are important vectors of antibiotic resistance and can be disseminated from hospital effluent to aquatic environments. This information is relevant to support management options aiming at the control of this public health problem.

  11. Roseomonas vinacea sp. nov., a Gram-negative coccobacillus isolated from a soil sample.

    Science.gov (United States)

    Zhang, Yu-Qin; Yu, Li-Yan; Wang, Dong; Liu, Hong-Yu; Sun, Cheng-Hang; Jiang, Wei; Zhang, Yue-Qin; Li, Wen-Jun

    2008-09-01

    Strain CPCC 100056(T), which was isolated from a soil sample collected from the Qinghai-Tibet plateau, China, was subjected to a polyphasic taxonomic study. The organism was coccobacillus-shaped, non-motile and formed vinaceous colonies on ISP2 agar medium. The respiratory quinone was ubiquinone-10. The major fatty acids were C(18:1)omega7c and C(16:1)omega7c and/or C(16:1)omega6c. The G+C content of the genomic DNA was 67.3 mol%. A comparison of sequences in GenBank revealed that strain CPCC 100056(T) exhibited highest 16S rRNA gene sequence similarity (84.5-95.5%) with Roseomonas species. Strain CPCC 100056(T) could be distinguished from all Roseomonas species with validly published names by differences in phenotypic and genotypic properties. In view of the combined phenotypic, chemotaxonomic and phylogenetic data, strain CPCC 100056(T) should be classified as a representative of a novel species in the genus Roseomonas, Roseomonas vinacea sp. nov.; the type strain is CPCC 100056(T) (=KCTC 22045(T) =CCM 7468(T)).

  12. How to identify Raoultella spp. including R. ornithinolytica isolates negative for ornithine decarboxylase? The reliability of the chromosomal bla gene.

    Science.gov (United States)

    Walckenaer, Estelle; Leflon-Guibout, Véronique; Nicolas-Chanoine, Marie-Hélène

    2008-12-01

    Although Raoultella planticola and Raoultella ornithinolytica were described more than 20 years ago, identifying them remains difficult. The reliability of the chromosomal bla gene for this identification was evaluated in comparison with that of the 16S rDNA and rpoB genes in 35 Raoultella strains from different origins. Of the 26 strains previously identified as R. planticola by biochemical tests alone or in association with molecular methods, 21 harboured a bla gene with 99.8% identity with the bla gene of two reference R. ornithinolytica strains (bla(ORN) gene) and 5 harboured a bla gene with 99.2% identity with the bla gene of two reference R. planticola strains (bla(PLA) gene). The 9 isolates previously identified as R. ornithinolytica harboured a bla(ORN) gene. The bla gene-based identification was confirmed by 16S rDNA and rpoB sequencing. The 21 isolates newly identified as R. ornithinolytica had a test negative for ornithine decarboxylase (ODC). Molecular experiments suggested one copy of ODC-encoding gene in both ODC-negative R. ornithinolytica and R. planticola strains and two copies in ODC-positive R. orninthinolytica strains. Analysis of the 35 bla genes allowed us (i) to confirm an identity of only 94% between the bla genes of the two Raoultella species while this identity was > 98% for rpoB and > 99% for 16S rDNA genes and (ii) to develop and successfully apply a bla PCR RFLP assay for Raoultella spp. identification. Overall, this study allowed us to discover ODC-negative R. ornithinolytica and to provide a reliable Raoultella identification method widely available as not requiring sequencing equipment.

  13. Novel urease-negative Helicobacter sp. 'H. enhydrae sp. nov.' isolated from inflamed gastric tissue of southern sea otters.

    Science.gov (United States)

    Shen, Zeli; Batac, Francesca; Mannion, Anthony; Miller, Melissa A; Bakthavatchalu, Vasudevan; Ho, Calvin; Manning, Sean; Paster, Bruce J; Fox, James G

    2017-02-08

    A total of 31 sea otters Enhydra lutris nereis found dead or moribund (and then euthanized) were necropsied in California, USA. Stomach biopsies were collected and transected with equal portions frozen or placed in formalin and analyzed histologically and screened for Helicobacter spp. in gastric tissue. Helicobacter spp. were isolated from 9 sea otters (29%); 58% (18 of 31) animals were positive for helicobacter by PCR. The Helicobacter sp. was catalase- and oxidase-positive and urease-negative. By electron microscopy, the Helicobacter sp. had lateral and polar sheathed flagella and had a slightly curved rod morphology. 16S and 23S rRNA sequence analyses of all 'H. enhydrae' isolates had similar sequences, which clustered as a novel Helicobacter sp. closely related to H. mustelae (96-97%). The genome sequence of isolate MIT 01-6242 was assembled into a single ~1.6 Mb long contig with a 40.8% G+C content. The annotated genome contained 1699 protein-coding sequences and 43 RNAs, including 65 genes homologous to known Helicobacter spp. and Campylobacter spp. virulence factors. Histological changes in the gastric tissues extended from mild cystic degeneration of gastric glands to severe mucosal erosions and ulcers. Silver stains of infected tissues demonstrated slightly curved bacterial rods at the periphery of the gastric ulcers and on the epithelial surface of glands. The underlying mucosa and submucosa were infiltrated by low numbers of neutrophils, macrophages, and lymphocytes, with occasional lymphoid aggregates and well-defined lymphoid follicles. This is the second novel Helicobacter sp., which we have named 'H. enhydrae', isolated from inflamed stomachs of mustelids, the first being H. mustelae from a ferret.

  14. Multiplex Real-Time PCR Assay for Rapid Detection of Methicillin-Resistant Staphylococci Directly from Positive Blood Cultures

    OpenAIRE

    Wang, Hye-young; Kim, Sunghyun; Kim, Jungho; Park, Soon-Deok; Uh, Young; Lee, Hyeyoung

    2014-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is the most prevalent cause of bloodstream infections (BSIs) and is recognized as a major nosocomial pathogen. This study aimed to evaluate a newly designed multiplex real-time PCR assay capable of the simultaneous detection of mecA, S. aureus, and coagulase-negative staphylococci (CoNS) in blood culture specimens. The Real-MRSA and Real-MRCoNS multiplex real-time PCR assays (M&D, Republic of Korea) use the TaqMan probes 16S rRNA for Staphylo...

  15. Perfil de contaminação por Staphylococcus e suas enterotoxinas e monitorização das condições de higiene em uma linha de produção de queijo de coalho Contamination profile for staphylococci and its enterotoxins and monitorization of the conditions of hygiene in a ‘coalho' cheese production line

    Directory of Open Access Journals (Sweden)

    Maria de Fatima Borges

    2008-08-01

    Full Text Available Avaliou-se o perfil de contaminação por Staphylococcus e suas enterotoxinas e monitorou-se as condições de higiene em uma linha de produção de queijo de coalho por meio da técnica de bioluminescência (ATP. A população de Staphylococcus sp. variou de This research aimed to evaluate the contamination by staphylococci and its enterotoxins as well as to monitor the conditions of hygiene from a coalho cheese production line, using ATP bioluminescence assay. Staphylococcus sp. population varied from <1CFU mL-1, in pasteurized milk to 1.5 x 107CFU mL-1, in raw milk, whereas coagulase-positive staphylococci count ranged from <1CFU mL-1, in pasteurized milk to 5.0 x 106CFU mL-1 in raw milk. Coagulase-positive staphylococci were detected in 100% (25/25 of the raw milk samples and in 8% (2/25 of cheese samples. Twelve Staphylococcus species were identified within the selected 68 isolates, being nine negative and three positive for coagulase. Raw milk samples showed a high rate of coagulase-positive, being S. aureus the most common, whereas other product samples and equipment surfaces, pieces of furniture, utensils and manipulator gloves samples presented a high frequency of coagulase-negative and low frequency of coagulase-positive. Staphylococcal enterotoxin was detected in 20% of the raw milk samples and therefore in pasteurized milk, curd and cheese. ATP measurement permitted to assess the effectiveness of the surfaces cleaning, being considered adequate in 62.1% (36/95, "alert state" in 23.2% (22/95 and inadequate in 14.7% (14/95 of surfaces evaluated. Detection of staphylococci species with enterotoxigenic potential as well as enterotoxin presence reveal dissemination of contamination at the "coalho" cheese production line, possibly due inappropriate Good Manufacturing Practices (GMP from the initial milking step until the final cheese production.

  16. Micro-organisms isolated from cadaveric samples of allograft musculoskeletal tissue.

    Science.gov (United States)

    Varettas, Kerry

    2013-12-01

    Allograft musculoskeletal tissue is commonly used in orthopaedic surgical procedures. Cadaveric donors of musculoskeletal tissue supply multiple allografts such as tendons, ligaments and bone. The microbiology laboratory of the South Eastern Area Laboratory Services (SEALS, Australia) has cultured cadaveric allograft musculoskeletal tissue samples for bacterial and fungal isolates since 2006. This study will retrospectively review the micro-organisms isolated over a 6-year period, 2006-2011. Swab and tissue samples were received for bioburden testing and were inoculated onto agar and/or broth culture media. Growth was obtained from 25.1 % of cadaveric allograft musculoskeletal tissue samples received. The predominant organisms isolated were coagulase-negative staphylococci and coliforms, with the heaviest bioburden recovered from the hemipelvis. The rate of bacterial and fungal isolates from cadaveric allograft musculoskeletal tissue samples is higher than that from living donors. The type of organism isolated may influence the suitability of the allograft for transplant.

  17. The most common isolated microorganisms and its clinical manifestation in hospitalised women suffering from urogenital diseases in Latvia

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    Anatolijs Naum and #269;iks

    2015-08-01

    Full Text Available Background: The monitoring of genital tract infections, especially sexually transmitted infections, is a vital part of health priority in many countries. One of the most common vaginal infections in women of childbearing age is bacterial vaginosis. Methods: This is a laboratory based retrospective study. We analysed 774 patients' cases (age range 18-35 years. Patients were subcategorized into three groups: patients with inflammation in genital tract, patients without inflammatory processes in genital tract, threatened abortion women. Laboratory tests also were divided depending on the type of examination. Results: There were 49 patients in the first group, 60 women in the second group and 236 pregnant women with threatened abortion in the third group. Gram negative bacteria (Escherichia coli and Enterococcus spp. were the most common isolated bacteria in the first and second group. Coagulase-negative staphylococci were predominant in the third group. Streptococcus agalactiae was isolated in 11.8 % cases. In our study, Trichomonas vaginalis infection was found in 1.2%. Conclusions: The most common isolated bacteria were Coagulase-negative staphylococci and gram negative bacteria. Ureaplasma urealyticum was detected in a decisive majority (38.9% of Mycoplasma species infections. In the first patients group and ndash; women with inflammatory process in genital tract were not found 3 bacteria, opposite to the second and third group. [Int J Reprod Contracept Obstet Gynecol 2015; 4(4.000: 959-962

  18. Multi-locus sequence typing (MLST) of non-fermentative Gram-negative bacilli isolated from bloodstream infections in southern Poland.

    Science.gov (United States)

    Chmielarczyk, Agnieszka; Pobiega, Monika; Romaniszyn, Dorota; Wójkowska-Mach, Jadwiga

    2017-09-22

    Non-fermentative Gram-negative bacilli are now one of the most important causes of severe infections in Polish hospitals. Acinetobacter species are serious concern because of the high prevalence of multi-drug resistance among strains. Resistance profiles for 53 Gram-negative non-fermentative blood isolates were done. MLST was carried out using 44 strains representing the most commonly isolated species: A. baumannii, P. aeruginosa, and S. maltophilia. MLST revealed that all 22 A. baumannii belonged to sequence type (ST) 2. The P. aeruginosa isolates belonged to 10 different STs. Four S. maltophilia isolates matched STs present in the database (ST4, ST15, ST116, ST142), seven isolates showing novel sequence types. Among P. aeruginosa and S. maltophilia PFGE confirmed the genetical variety of strains.

  19. Prevalence and antimicrobial susceptibilities of bacteria isolated from blood cultures of hospitalized patients in the United States in 2002

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    Thornsberry Clyde

    2004-05-01

    Full Text Available Abstract Background Bloodstream infections are associated with significant patient morbidity and mortality. Antimicrobial susceptibility patterns should guide the choice of empiric antimicrobial regimens for patients with bacteremia. Methods From January to December of 2002, 82,569 bacterial blood culture isolates were reported to The Surveillance Network (TSN Database-USA by 268 laboratories. Susceptibility to relevant antibiotic compounds was analyzed using National Committee for Clinical Laboratory Standards guidelines. Results Coagulase-negative staphylococci (42.0%, Staphylococcus aureus (16.5%, Enterococcus faecalis (8.3%, Escherichia coli (7.2%, Klebsiella pneumoniae (3.6%, and Enterococcus faecium (3.5% were the most frequently isolated bacteria from blood cultures, collectively accounting for >80% of isolates. In vitro susceptibility to expanded-spectrum β-lactams such as ceftriaxone were high for oxacillin-susceptible coagulase-negative staphylococci (98.7%, oxacillin-susceptible S. aureus (99.8%, E. coli (97.3%, K. pneumoniae (93.3%, and Streptococcus pneumoniae (97.2%. Susceptibilities to fluoroquinolones were variable for K. pneumoniae (90.3–91.4%, E. coli (86.0–86.7%, oxacillin-susceptible S. aureus (84.0–89.4%, oxacillin-susceptible coagulase-negative staphylococci (72.7–82.7%, E. faecalis (52.1%, and E. faecium (11.3%. Combinations of antimicrobials are often prescribed as empiric therapy for bacteremia. Susceptibilities of all blood culture isolates to one or both agents in combinations of ceftriaxone, ceftazdime, cefepime, piperacillin-tazobactam or ciprofloxacin plus gentamicin were consistent (range, 74.8–76.3% but lower than similar β-lactam or ciprofloxacin combinations with vancomycin (range, 93.5–96.6%. Conclusion Ongoing surveillance for antimicrobial susceptibility remains essential, and will enhance efforts to identify resistance and attempt to limit its spread.

  20. Antimicrobial activity of ceftobiprole against Gram-negative and Gram-positive pathogens: results from INVITA-A-CEFTO Brazilian study

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    Rosângela Ferraz Cereda

    2011-08-01

    Full Text Available Ceftobiprole is a broad-spectrum cephalosporin with potent activity against staphylococci, including those resistant to oxacillin, as well as against most Gram-negative bacilli including Pseudomonas aeruginosa. In this study, the in vitro activity of ceftobiprole and comparator agents was tested against bacterial isolates recently collected from Brazilian private hospitals. A total of 336 unique bacterial isolates were collected from hospitalized patients between February 2008 and August 2009. Each hospital was asked to submit 100 single bacterial isolates responsible for causing blood, lower respiratory tract or skin and soft tissue infections. Bacterial identification was confirmed and antimicrobial susceptibility testing was performed using CLSI microdilution method at a central laboratory. The CLSI M100-S21 (2011 was used for interpretation of the antimicrobial susceptibility results. Among the 336 pathogens collected, 255 (75.9% were Gram-negative bacilli and 81 (24.1% were Gram-positive cocci. Although ceftobiprole MIC50 values for oxacillin resistant strains were two-fold higher than for methicillin susceptible S. aureus, ceftobiprole inhibited 100% of tested S. aureus at MICs 6 µg/mL for both species. Our results showed that ceftobiprole has potent activity against staphylococci and E. faecalis, which was superior to that of vancomycin. Our data also indicates that ceftobiprole demonstrated potency comparable to that of cefepime and ceftazidime against key Gram-negative species.

  1. Antimicrobial activity of ceftobiprole against gram-negative and gram-positive pathogens: results from INVITA-A-CEFTO Brazilian study.

    Science.gov (United States)

    Cereda, Rosângela Ferraz; Azevedo, Heber Dias; Girardello, Raquel; Xavier, Danilo Elias; Gales, Ana C

    2011-01-01

    Ceftobiprole is a broad-spectrum cephalosporin with potent activity against staphylococci, including those resistant to oxacillin, as well as against most gram-negative bacilli including Pseudomonas aeruginosa. In this study, the in vitro activity of ceftobiprole and comparator agents was tested against bacterial isolates recently collected from Brazilian private hospitals. A total of 336 unique bacterial isolates were collected from hospitalized patients between February 2008 and August 2009. Each hospital was asked to submit 100 single bacterial isolates responsible for causing blood, lower respiratory tract or skin and soft tissue infections. Bacterial identification was confirmed and antimicrobial susceptibility testing was performed using CLSI microdilution method at a central laboratory. The CLSI M100-S21 (2011) was used for interpretation of the antimicrobial susceptibility results. Among the 336 pathogens collected, 255 (75.9%) were gram-negative bacilli and 81 (24.1%) were gram-positive cocci. Although ceftobiprole MIC50 values for oxacillin resistant strains were two-fold higher than for methicillin susceptible S. aureus, ceftobiprole inhibited 100% of tested S. aureus at MICs ceftobiprole MIC50 > 6 µg/mL for both species. Our results showed that ceftobiprole has potent activity against staphylococci and E. faecalis, which was superior to that of vancomycin. Our data also indicates that ceftobiprole demonstrated potency comparable to that of cefepime and ceftazidime against key gram-negative species.

  2. Staphylococci in cattle and buffaloes with mastitis in Dakahlia Governorate, Egypt.

    Science.gov (United States)

    El-Ashker, Maged; Gwida, Mayada; Tomaso, Herbert; Monecke, Stefan; Ehricht, Ralf; El-Gohary, Fatma; Hotzel, Helmut

    2015-11-01

    The aim of this study was to provide the first detailed insight into the population structure of Staphylococcus aureus in one modern dairy farm (Gamasa) and several household cows and buffaloes in Dakahlia Governorate, Egypt. Eight hundred seventy-two quarter milk samples of 218 dairy cattle and buffaloes with clinical and subclinical mastitis were investigated. Bacteria were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and staphylococci were further characterized by DNA sequencing of 16S rRNA genes and microarray analysis. Staphylococcus aureus was present in 5.6% of all collected samples, whereas methicillin-resistant S. aureus (MRSA) represented 24.5% of all identified S. aureus (12/49). Six clonal complexes (CC) of S. aureus were detected. Staphylococcus aureus CC398 (ST291/813)-MSSA (methicillin-susceptible S. aureus) was identified frequently in the Gamasa farm in addition to a few CC5-MRSA-V isolates. However, a small number of different isolates of S. aureus were found in household cattle and buffaloes harboring different CC. The presence of these genotypes of S. aureus in milk might indicate a public health hazard, because all of these CC have previously been isolated from human patients. Thus, a recommendation was given to the owner of the dairy farm to review the hygiene regimen on the farm. In perspective, further investigation regarding S. aureus screening of all lactating cows and personnel on the farm is warranted.

  3. Antibiotic resistance and enterotoxin genes in Staphylococcus sp. isolates from polluted water in Southern Brazil

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    ANA P. BASSO

    2014-12-01

    Full Text Available The aim of this study was to evaluate the species distribution, antibiotic-resistance profile and presence of enterotoxin (SE genes in staphylococci isolated from the Dilúvio stream in South Brazil. Eighty-eight staphylococci were identified, 93.18% were identified as coagulase-negative (CNS and 6.82% coagulase-positive (CPS. Fourteen Staphylococcus species were detected and the most frequently were Staphylococcus cohnii (30.48% and S. haemolyticus (21.95%. Resistance to erythromycin was verified in 37.50% of the strains, followed by 27.27% to penicillin, 12.50% to clindamycin, 6.81% to trimethoprim-sulfamethoxazole, 5.68% to chloramphenicol and 2.27% to norfloxacin. None of the investigated strains showed gentamicin and ciprofloxacin resistance. The strains were tested for the presence of sea, seb, sec, sed and see genes by PCR and only CNS strains (43.18% showed positive results to one or more SE genes. The scientific importance of our results is due to the lack of data about these topics in polluted waters in Brazil. In conclusion, polluted waters from the Dilúvio stream may constitute a reservoir for disseminating antibiotic-resistance and enterotoxin into the community. In addition, the detection of staphylococci in the polluted waters of the Dilúvio stream indicated a situation of environmental contamination and poor sanitation conditions.

  4. DETERMINACIÓN DEL GEN aac(6´-aph(2´´ ASOCIADO CON RESISTENCIA A AMINOGLUCÓSIDOS EN CEPAS DE STAPHYLOCOCCUS COAGULASA NEGATIVA EN UNA UNIDAD NEONATAL EN BOGOTÁ Detection of gene aac(6´-aph(2´´ in associated with aminoglucosides resistance in Staphylococcus coagulase-negative strains isolates from one neonatal unit in Bogota

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    Liliana Muñoz M

    2009-01-01

    Full Text Available Antecedentes. La causa más común de resistencia a antibióticos aminoglucósidos en bacterias Gram positivas, especialmente en S. epidermidis, es la enzima modificante AAC(6’-APH(2", capaz de acetilar y fosforilar un amplio rango de antibióticos. Objetivo. Determinar la presencia del gen aac(6’-aph(2" en cepas de Staphylococcus coagulasa negativa aisladas en infecciones neonatales, e investigar la concordancia con las pruebas de sensibilidad in-vitro. Material y métodos. Se determinó la presencia del gen aac(6’-aph(2" en 63 cepas de estafilococos coagulasa negativa provenientes de hemocultivos y puntas de catéter, de la Unidad de Neonatología del Instituto Materno Infantil, en Bogotá. Resultados. Staphylococcus epidermidis fue el germen más frecuente y el gen aac(6’-aph(2" estaba presente en 55 (87,3% cepas; de éstas, 42 (73,4% cepas provenían de hemocultivos y 13 (23,6% cepas de punta de catéter. La susceptibilidad a gentamicina se determinó mediante concentración inhibitoria mínima y para amikacina por difusión en disco. No se encontró asociación estadísticamente significativa entre la presencia del gen aac(6’-aph(2" y la resistencia a gentamicina y amikacina. Conclusión. La presencia del gen aac(6’-aph(2" de resistencia a aminoglucósidos es muy alta en cepas de Staphylococcus epidermidis. Diferencias en la expresión del gen pueden explicar parcialmente la falta de consistencia con las pruebas de susceptibilidad utilizadas en la clínica.Background. The most common cause of resistance to aminoglucosides is the acetilation and phosphorilation of the antibiotic by the enzyme AAC(6’APH(2". Objetive. To determine the presence of the gene aac(6’-aph(2" in strains of coagulase-negative Staphylococci isolated from infected neonates and to investigate the concordance with the susceptibility tests in-vitro. Materials and methods. the aac(6’-aph(2" gene was determined in 63 coagulase-negative Staphylococci strains

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

    Science.gov (United States)

    Andrasevic, Arjana Tambic; Dowzicky, Michael J

    2012-02-01

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

  6. A survey of five broad-host-range plasmids in gram-negative bacilli isolated from patients.

    Science.gov (United States)

    Zhang, Guo-qiang; Yao, Yi-hui; Yu, Xiao-lu; Niu, Jian-jun

    2014-07-01

    To learn the prevalence of the primary classical broad-host-range (BHR) IncA/C, IncN, IncP, IncQ, and IncW plasmids in dominant gram-negative bacilli from inpatients in a teaching hospital in southern China. A multiplex polymerase chain reaction based on the replicons of BHR IncA/C, IncN, IncP, IncQ, and IncW plasmids was developed and used to determine these BHR plasmids. The difference in prevalence rates among the different species from three specimens was evaluated by a binary logistic regression model and the differences between multidrug-resistant organisms (MDRO) and non-MDRO were assessed using a chi-square test. The average positive detection percentages of the replicons were 4.3%, 3.7%, 3.0%, 2.6%, and 1.9%, respectively, for IncN, IncP, IncQ, IncW, and IncA/C in descending order. The distribution of all five BHR plasmids did not differ significantly between specimens collected from wounds and urine, although both were significantly higher than those of sputum. The prevalence rates of all five BHR plasmids in MDROs were significantly higher than those in non-MDRO for Enterobacteriaceae; however, no significant difference was seen in non-fermenting gram-negative bacilli (NFGNB). BHR IncA/C, IncN, IncP, IncQ, and IncW plasmids, which occur more often in bacilli from wound and urine specimens than those of sputum, are widespread in Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Enterobacter cloacae, Pseudomonas aeruginosa, and Acinetobacter baumannii strains isolated from inpatients. The prevalence rates in MDRO are higher than those in non-MDRO for Enterobacteriaceae but not significantly different for NFGNB. Copyright © 2014 Elsevier Inc. All rights reserved.

  7. Duplex real-time PCR assay for rapid identification of Staphylococcus aureus isolates from dairy cow milk.

    Science.gov (United States)

    Pilla, Rachel; Snel, Gustavo G M; Malvisi, Michela; Piccinini, Renata

    2013-05-01

    Staphylococcus aureus isolates from dairy cow mastitis are not always consistent with the characteristic morphology described, and molecular investigation is often needed. The aim of the study was to develop a duplex real-time PCR assay for rapid identification of Staph. aureus isolates, targeting both nuc and Sa442. Overall, 140 isolates collected from dairy cow mastitis in 90 different herds, were tested. All strains had been identified using morphological and biochemical characteristics. DNA from each strain was amplified in real-time PCR assay, to detect nuc or Sa442. Thereafter, a duplex real-time PCR assay was performed, and specificity of the amplified products was assessed by high resolution melting curve analysis. Out of 124 Staph. aureus isolates, 33 did not show the typical morphology or enzymic activity; in 118 strains, the two melt-curve peaks consistent with nuc and Sa442 were revealed, while 2 isolates showed only the peak consistent with Sa442. Four isolates bacteriologically identified as Staph. aureus, were PCR-negative and were further identified as Staph. pseudintermedius by sequencing. Staph. pseudintermedius and coagulase-negative staphylococci did not carry nuc or Sa442. The results showed the correct identification of all isolates, comprehending also coagulase-or nuc-negative Staph. aureus, while other coagulase-positive Staphylococci were correctly identified as non-Staph. aureus. Both sensitivity and specificity were 100%. High resolution melting analysis allowed easy detection of unspecific products. Finally, the duplex real-time PCR was applied directly to 40 milk samples, to detect infected mammary quarters. The assay confirmed the results of bacteriological analysis, on Staph. aureus-positive or-negative samples. Therefore, the proposed duplex real-time PCR could be used in laboratory routine as a cost-effective and powerful tool for high-throughput identification of atypical Staph. aureus isolates causing dairy cow mastitis. Also, it

  8. A surveillance study of antimicrobial resistance of gram-negative bacteria isolated from intensive care units in eight hospitals in Turkey.

    Science.gov (United States)

    Günseren, F; Mamikoğlu, L; Oztürk, S; Yücesoy, M; Biberoğlu, K; Yuluğ, N; Doğanay, M; Sümerkan, B; Kocagöz, S; Unal, S; Cetin, S; Calangu, S; Köksal, I; Leblebicioğlu, H; Günaydin, M

    1999-03-01

    This study was carried out with the participation of eight hospitals in Turkey to determine the frequency of gram-negative bacteria isolated in intensive care units (ICU) and to compare their resistance rates to selected antibiotics. Aerobic gram-negative bacteria isolated from ICUs during 1996 were studied. Antibiotic susceptibilities to imipenem, ceftazidime, ceftazidime-clavulanate, ceftriaxone, cefotaxime, cefepime, cefodizime, cefuroxime, piperacillin/tazobactam, amoxycillin-clavulanate, gentamicin, amikacin and ciprofloxacin were determined by Etest. A total of 748 isolates were obtained from 547 patients. The majority of organisms were isolated from the respiratory (38.8%) and urinary tracts (30.9%). Pseudomonas spp. were the most frequently isolated gram-negative species (26.8%), followed by Klebsiella spp. (26.2%). Escherichia coli, Acinetobacter spp. and Enterobacter spp. were the other commonly isolated organisms. High resistance rates were observed for all antibiotics studied. Imipenem appeared to be the most active agent against the majority of isolates. Although resistance rates exceeded 50%, ciprofloxacin, cefepime and amikacin were found to be relatively effective. Extended-spectrum beta-lactamase (ESBL) production appeared to be a major mechanism of resistance to beta-lactam antibiotics. In contrast to ceftazidime-clavulanate, piperacillin/tazobactam showed poor activity against organisms thought to produce ESBL, suggesting the presence of an enzyme resistant to tazobactam action. This study has yielded high rates of resistance in aerobic gram-negative isolates from ICUs in Turkey. High resistance rates to all the other antibacterials studied leave imipenem as the only reliable agent for the empirical treatment of ICU infections in Turkey.

  9. Antibiotic resistance of gram-negative bacilli isolated from pediatric patients with nosocomial bloodstream infections in a Mexican tertiary care hospital.

    Science.gov (United States)

    Ares, Miguel Ángel; Alcántar-Curiel, Maria Dolores; Jiménez-Galicia, César; Rios-Sarabia, Nora; Pacheco, Sabino; De la Cruz, Miguel Ángel

    2013-01-01

    Gram-negative bacilli are the most common bacteria causing nosocomial bloodstream infections (NBSIs) in Latin American countries. The antibiotic resistance profiles of Gram-negative bacilli isolated from blood cultures in pediatric patients with NBSIs over a 3-year period in a tertiary care pediatric hospital in Mexico City were determined using the VITEK-2 system. Sixteen antibiotics were tested to ascertain the resistance rate and the minimum inhibitory concentration using the Clinical Laboratory Standards Institute (CLSI) broth micro-dilution method as a reference. A total of 931 isolates were recovered from 847 clinically significant episodes of NBSI. Of these, 477 (51.2%) were caused by Gram-negative bacilli. The most common Gram-negative bacilli found were Klebsiella pneumoniae (30.4%), Escherichia coli (18.9%), Enterobacter cloacae (15.1%), Pseudomonas aeruginosa (9.9%), and Acinetobacter baumannii (4.6%). More than 45 and 60% of the K. pneumoniae and E. coli isolates, respectively, were resistant to cephalosporins, and 64% of the E. coli isolates were resistant to fluoroquinolones. A. baumannii exhibited low rates of resistance to antibiotics tested. In the E. cloacae and P. aeruginosa isolates, no rates of resistance higher than 38% were observed. In this study, we found that the proportion of NBSIs due to antibiotic-resistant organisms is increasing in a tertiary care pediatric hospital of Mexico.

  10. Resistance among Gram-negative ESKAPE pathogens isolated from hospitalized patients with intra-abdominal and urinary tract infections in Latin American countries: SMART 2013-2015

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    James A. Karlowsky

    Full Text Available ABSTRACT Gram-negative ESKAPE pathogens (Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species are important etiologic agents of nosocomial infection that are frequently resistant to broad-spectrum antimicrobial agents. Gram-negative ESKAPE pathogens were collected from hospitalized patients in 11 Latin American countries from 2013 to 2015 as part of the Study for Monitoring Antimicrobial Resistance Trends (SMART global surveillance program. In total, 2113 isolates from intra-abdominal infections (IAI and 970 isolates from urinary tract infections (UTI were tested against antimicrobial agents using standardized CLSI broth microdilution methodology. Of the agents tested, amikacin demonstrated the highest rates of susceptibility (% for K. pneumoniae (92.2, 92.3, Enterobacter spp. (97.5, 92.1, and P. aeruginosa (85.3, 75.2 isolates from both IAI and UTI, respectively. Ertapenem (68.5, 62.6 and imipenem (79.2, 75.9 showed substantially higher rates of susceptibility (% than other β-lactams, including piperacillin-tazobactam (35.9, 37.4 against ESBL-positive isolates of K. pneumoniae from IAI and UTI, respectively. Rates of susceptibility to all agents tested against A. baumannii were ≤30.9%. Gram-negative ESKAPE pathogens isolated from Latin America demonstrated compromised in vitro susceptibility to commonly prescribed broad-spectrum, parenteral antimicrobial agents. Continued surveillance is warranted. New antimicrobial agents with potent activity against Gram-negative ESKAPE pathogens are urgently needed.

  11. Antimicrobial susceptibility trends among gram-positive and -negative clinical isolates collected between 2005 and 2012 in Mexico: results from the Tigecycline Evaluation and Surveillance Trial

    OpenAIRE

    Morfin-Otero, Rayo; Noriega, Eduardo Rodriguez; Dowzicky, Michael J.

    2015-01-01

    Background The Tigecycline Evaluation and Surveillance Trial (T.E.S.T) is a global antimicrobial surveillance study of both gram-positive and gram-negative organisms. This report presents data on antimicrobial susceptibility among organisms collected in Mexico between 2005 and 2012 as part of T.E.S.T., and compares rates between 2005–2007 and 2008–2012. Method Each center in Mexico submitted at least 200 isolates per collection year; including 65 gram-positive isolates and 135 gram-negative i...

  12. Differentiation of Staphylococci from Iberian dry fermented sausages by protein fingerprinting.

    Science.gov (United States)

    Benito, María J; Serradilla, Manuel J; Martín, Alberto; Aranda, Emilio; Hernández, Alejandro; Córdoba, María G

    2008-08-01

    The Staphylococci populations in different types of Iberian dry fermented sausages from central-west Spain were identified. A simple electrophoretic method of whole-cell proteins and extracellular protein profiling was evaluated for speed of identification. This study was correlated with a 16S rRNA gene sequence analysis and biochemical identification by API Staph. A total of 81 isolates were identified by SDS-PAGE of the whole-cell proteins. These showed stable profiles in the range 99-14kDa that were clearly different for the different species, and were grouped into clusters together with the profiles of the eight reference strains. SDS-PAGE of the extracellular protein extracts provided additional characteristic banding patterns for the characterization of the Staphylococcus species present. The whole-cell SDS-PAGE showed that the predominant species was Staphylococcus saprophyticus (61.7%) followed by Staphylococcus aureus (19.7%). The identifications were confirmed by the 16S rRNA gene sequencing and by a BLAST search of the GenBank database. However, the API Staph biochemical identifications were frequently erroneous at the species level. In sum, SDS-PAGE analysis showed itself to be rapid and accurate in identifying the most commonly encountered Staphylococcus isolates in dry fermented sausages.

  13. Comprehensive Phylogenetic Analysis of Bovine Non-aureus Staphylococci Species Based on Whole-Genome Sequencing

    Science.gov (United States)

    Naushad, Sohail; Barkema, Herman W.; Luby, Christopher; Condas, Larissa A. Z.; Nobrega, Diego B.; Carson, Domonique A.; De Buck, Jeroen

    2016-01-01

    Non-aureus staphylococci (NAS), a heterogeneous group of a large number of species and subspecies, are the most frequently isolated pathogens from intramammary infections in dairy cattle. Phylogenetic relationships among bovine NAS species are controversial and have mostly been determined based on single-gene trees. Herein, we analyzed phylogeny of bovine NAS species using whole-genome sequencing (WGS) of 441 distinct isolates. In addition, evolutionary relationships among bovine NAS were estimated from multilocus data of 16S rRNA, hsp60, rpoB, sodA, and tuf genes and sequences from these and numerous other single genes/proteins. All phylogenies were created with FastTree, Maximum-Likelihood, Maximum-Parsimony, and Neighbor-Joining methods. Regardless of methodology, WGS-trees clearly separated bovine NAS species into five monophyletic coherent clades. Furthermore, there were consistent interspecies relationships within clades in all WGS phylogenetic reconstructions. Except for the Maximum-Parsimony tree, multilocus data analysis similarly produced five clades. There were large variations in determining clades and interspecies relationships in single gene/protein trees, under different methods of tree constructions, highlighting limitations of using single genes for determining bovine NAS phylogeny. However, based on WGS data, we established a robust phylogeny of bovine NAS species, unaffected by method or model of evolutionary reconstructions. Therefore, it is now possible to determine associations between phylogeny and many biological traits, such as virulence, antimicrobial resistance, environmental niche, geographical distribution, and host specificity. PMID:28066335

  14. Prevalence and antimicrobial susceptibility of non-fermenting Gram-negative bacilli isolated from clinical samples at a tertiary care hospital.

    Science.gov (United States)

    Argenta, Anne de Rossi; Fuentefria, Daiane Bopp; Sobottka, Andréa Michel

    2017-01-01

    We compared the prevalence and antimicrobial susceptibility of non-fermenting gram-negative bacilli (NFGNB) isolated from clinical samples at a Brazilian tertiary care hospital in 2008 and 2013. Collected data included patient's name, age, sex, inpatient unit, laboratory record number, type of biological material, culture test result, and antimicrobial susceptibility of isolated strains. Out of 19,112 culture tests analyzed, 926 (4.8%) were positive for NFGNB. Among these, 45.2% were metallo-beta-lactamase (MBL) producing strains. Between 2008 and 2013, the number of MBL-producing NFGNB isolates increased by 21.5%, which was accompanied by a consequent reduction in susceptibility to antimicrobials.

  15. [Microbial flora isolated from patient's conjunctiva previous to cataract surgery].

    Science.gov (United States)

    Barría von-B, Fernando; Chabouty, Henriette; Moreno, René; Ortiz, Freddy; Barría M, Fernando

    2015-04-01

    Endophtalmitis post cataract surgery is one of most feared and devastating complications resulting in serious consequences and an uncertain visual prognosis. Antimicrobial prophylaxis against endophtalmitis must be based on the best knowledge of conjuntival microbiota. To establish microbiological basis for the best antibiotic prophylaxis to prevent endophthalmitis in cataract surgery. A descriptive, cross-sectional, prospective study. A preoperative conjunctival sample was taken from the lower fornix of 118 pacients, sowing it immediately in culture media. Identification of growing colonies and susceptibility testing were performed by manual or automated methods. 106 (89.8%) of 118 preoperative cultures were positive. 159 bacteria were isolated in single or mixed flora, with 95% of Gram positive organisms. Staphylococci represented 76.1% of isolated bacteria, with 82.6% of coagulase-negative staphylococci (SCN) and 17.4% of Staphylococcus aureus. Forty two percent of SCN and 38% of S. aureus were methicillin resistan; both groups showed high susceptibility to tobramycin and fourth-generation fluoroquinolones. we recommend the use of topical tobramycin as pre-operative antimicrobial prophylaxis associated with povidone-iodine antisepsis. A fourth-generation quinolone is recommended when there is risk of infection.

  16. In vitro activity of fosfomycin combined with rifampin, pefloxacin and imipenem against staphylococci: a study by the time-kill curve method.

    Science.gov (United States)

    Quentin, C; Saivin, S; Lafferriere, C; Noury, P; Bebear, C

    1987-01-01

    The in vitro activity of fosfomycin alone and in combination with rifampin, pefloxacin and imipenem was studied by the time-kill method against staphylococci. Fosfomycin, pefloxacin and imipenem used at concentrations within the therapeutic range, exerted a bactericidal effect, whereas rifampin acted as a bacteriostatic drug. The combination of fosfomycin and rifampin was found to be antagonistic against rifampin-susceptible strains and indifferent for rifampin-resistant isolates. Fosfomycin combined with pefloxacin usually produced an indifferent effect. The interaction between fosfomycin and imipenem was mainly indifferent but synergism occurred with methicillin-resistant strains and antagonism was observed for one methicillin-susceptible isolate of Staphylococcus aureus.

  17. Genetic manipulation of Staphylococci – Breaking through the barrier.

    Directory of Open Access Journals (Sweden)

    Ian R Monk

    2012-04-01

    Full Text Available Most strains of Staphylococcus aureus and Staphylococcus epidermidis possess a strong restriction barrier that hinders exchange of DNA. Recently, major advances have been made in identifying and characterizing the restriction-modification systems involved. In particular a novel type IV restriction enzyme that recognizes cytosine methylated DNA has been shown to be the major barrier to transfer of plasmid DNA from Escherichia coli into S. aureus and S. epidermidis. One or sometimes two type I restriction-modification systems provide an additional barrier. Here we review the recent advances in understanding of restriction systems in staphylococci and highlight how this has been exploited to improve our ability to manipulate genetically previously untransformable strains.

  18. Presence of environmental coagulase-positive staphylococci, their clonal relationship, resistance factors and ability to form biofilm.

    Science.gov (United States)

    Velázquez-Guadarrama, Norma; Olivares-Cervantes, Alma L; Salinas, Eva; Martínez, Leticia; Escorcia, Magdalena; Oropeza, Ricardo; Rosas, Irma

    Coagulase-positive staphylococci (CoPS) are opportunistic pathogens carrying various mechanisms of resistance that have a large number of virulence factors, and whose ability to induce illness is associated with the host. This study aimed to investigate the presence of environmental coagulase-positive staphylococci, their susceptibility profile, clonal relationship and ability to form biofilm. The 16S rRNA genes from CoPS isolates were analyzed, and their antibiotic susceptibility was evaluated using the agar dilution method in accordance with Clinical and Laboratory Standards Institute guidelines. The clonal profile was obtained by pulsed-field gel electrophoresis (PFGE) and biofilm formation was measured by a crystal violet retention assay. A total of 72 Staphylococcus spp. strains were isolated from air, metal surfaces, and nostrils from humans, dogs, cats, and birds. Three species were identified: Staphylococcus aureus (17%), Staphylococcus intermedius (63%), and Staphylococcus pseudintermedius (21%). Ninety three percent (93%) of the strains were resistant to at least one of 13 tested antibiotics. S. pseudintermedius strains were the only resistant ones to methicillin while most of these isolates were multidrug-resistant, had significantly higher ability to form biofilm and PFGE grouped into seven different patterns, without showing clonal dispersion among animals and environmental isolates. This study suggests that dogs, cat, and air are environmental sources potentially carrying multidrug-resistant S. pseudintermedius, which survives in different environments through biofilm formation and multidrug resistance, characteristics that can be transmitted horizontally to other bacteria and exacerbate the problem of antibiotic resistance in humans. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  19. The prevalence of carriage of meticillin-resistant staphylococci by veterinary dermatology practice staff and their respective pets.

    Science.gov (United States)

    Morris, Daniel O; Boston, Raymond C; O'Shea, Kathleen; Rankin, Shelley C

    2010-08-01

    It has been shown that people and pets can harbour identical strains of meticillin-resistant (MR) staphylococci when they share an environment. Veterinary dermatology practitioners are a professional group with a high incidence of exposure to animals infected by Staphylococcus spp. The objective of this study was to assess the prevalence of carriage of MR Staphylococcus aureus (MRSA), MR S. pseudintermedius (MRSP) and MR S. schleiferi (MRSS) by veterinary dermatology practice staff and their personal pets. A swab technique and selective media were used to screen 171 veterinary dermatology practice staff and their respective pets (258 dogs and 160 cats). Samples were shipped by over-night carrier. Human subjects completed a 22-question survey of demographic and epidemiologic data relevant to staphylococcal transmission. The 171 human-source samples yielded six MRSA (3.5%), nine MRSP (5.3%) and four MRSS (2.3%) isolates, while 418 animal-source samples yielded eight MRSA (1.9%) 21 MRSP (5%), and two MRSS (0.5%) isolates. Concordant strains (genetically identical by pulsed-field gel electrophoresis) were isolated from human subjects and their respective pets in four of 171 (2.9%) households: MRSA from one person/two pets and MRSP from three people/three pets. In seven additional households (4.1%), concordant strains were isolated from only the pets: MRSA in two households and MRSP in five households. There were no demographic or epidemiologic factors statistically associated with either human or animal carriage of MR staphylococci, or with concordant carriage by person-pet or pet-pet pairs. Lack of statistical associations may reflect an underpowered study.

  20. Incidence of ESBL producers amongst Gram-negative bacilli isolated from intra-abdominal infections across India (based on SMART study, 2007 data).

    Science.gov (United States)

    Chaudhuri, B N; Rodrigues, C; Balaji, V; Iyer, R; Sekar, U; Wattal, Chand; Chitnis, D S; Dhole, T N; Joshi, Sangeeta

    2011-05-01

    This study was conducted in 9 centers spread over India from January 1 to December 31, 2007 to monitor in vitro susceptibility of Gram-negative bacilli to Group I carbapenem, ertapenem and other antimicrobials in intra-abdominal infections and to identify early changes in susceptibility pattern of community or hospital acquired organisms, with a focus on ESBL producers. Gram-negative bacilli isolated from intra-abdominal samples of patients with documented intra-abdominal infections were processed for identification by conventional/ automated methods and antimicrobial susceptibility by Micro-Scan (Siemens) MIC panel against 12 antimicrobials (3rd and 4th generation cephalosporins, Groups I and II carbapenems, amikacin, levofloxacin, amoxicillin-clavulanic acid and piperacillin-tazobactam). A total of 588 isolates were identified, of which 351 (60%) were E. coli and 114 (19%) were Klebsiella spp. 79% of E. coli and 70% of Klebsiella spp. were ESBL producers in general. 110 of E. coli and 35 of Klebsiella isolates were from community-acquired intra-abdominal infections. 80% of E. coli and 63% of Klebsiella isolates from community-acquired infections were ESBL producers, against 79% of E. coli and 73% of Klebsiella isolates from hospital-acquired infections. Amongst the ESBL-positive isolates of E. coli, 94% were susceptible in vitro to ertapenem, 96% to imipenem and 76% to piperacillin-tazobactam. For ESBL-positive isolates of Klebsiella spp., the corresponding figures were 80%, 94% and 59% respectively. The study showed a high incidence of ESBL-producers amongst Enterobacteriaceae isolates from intra-abdominal infections in both community-acquired and hospital-acquired settings across India. Ertapenem was comparable with imipenem against ESBL-positive E. coli isolates, while imipenem was more effective than ertapenem against ESBL-positive Klebsiella isolates.

  1. Spread of TEM, VIM, SHV, and CTX-M β-Lactamases in Imipenem-Resistant Gram-Negative Bacilli Isolated from Egyptian Hospitals.

    Science.gov (United States)

    Hamdy Mohammed, El Sayed; Elsadek Fakhr, Ahmed; Mohammed El Sayed, Hanan; Al Johery, Said Abd Elmohsen; Abdel Ghani Hassanein, Wesam

    2016-01-01

    Carbapenem-resistant Gram-negative bacilli resulting from β-lactamases have been reported to be an important cause of nosocomial infections and are a critical therapeutic problem worldwide. This study aimed to describe the prevalence of imipenem-resistant Gram-negative bacilli isolates and detection of bla VIM, bla TEM, bla SHV, bla CTX-M-1, and bla CTX-M-9 genes in these clinical isolates in Egyptian hospitals. The isolates were collected from various clinical samples, identified by conventional methods and confirmed by API 20E. Antibiotic susceptibility testing was determined by Kirby-Bauer technique and interpreted according to CLSI. Production of bla VIM, bla TEM, bla SHV, and bla CTX-M genes was done by polymerase chain reaction (PCR). Direct sequencing from PCR products was subsequently carried out to identify and confirm these β-lactamases genes. Out of 65 isolates, (46.1%) Escherichia coli, (26.2%) Klebsiella pneumoniae, and (10.7%) Pseudomonas aeruginosa were identified as the commonest Gram-negative bacilli. 33(50.8%) were imipenem-resistant isolates. 22 isolates (66.7%) carried bla VIM, 24(72.7%) had bla TEM, and 5(15%) showed bla SHV, while 12(36%), 6(18.2%), and 0(0.00%) harbored bla CTX-M-1, bla CTX-M-9, and bla CTX-M-8/25, respectively. There is a high occurrence of β-lactamase genes in clinical isolates and sequence analysis of amplified genes showed differences between multiple SNPs (single nucleotide polymorphism) sites in the same gene among local isolates in relation to published sequences.

  2. 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.

  3. Phenotypic and genotypic characterization of sorbitol-negative of slow-fermenting (suspected O157) Escherichia coli isolated from milk samples in Lombardy region

    NARCIS (Netherlands)

    Picozzi, C.; Foschino, R.; Heuvelink, A.E.; Beumer, R.R.

    2005-01-01

    Aims: To investigate phenotypic and genotypic aspects of sorbitol-negative or slow-fermenting Escherichia coli, suspected to belong to O157 serogroup, isolated in Italy. Methods and Results: Milk samples originating from goats and cows were screened for the presence of E. coli O157 with cultural met

  4. Family Rejection, Social Isolation, and Loneliness as Predictors of Negative Health Outcomes (Depression, Suicidal Ideation, and Sexual Risk Behavior) among Thai Male-to-Female Transgender Adolescents

    Science.gov (United States)

    Yadegarfard, Mohammadrasool; Meinhold-Bergmann, Mallika E.; Ho, Robert

    2014-01-01

    This study examined the influence of family rejection, social isolation, and loneliness on negative health outcomes among Thai male-to-female transgender adolescents. The sample consisted of 260 male respondents, of whom 129 (49.6%) were self-identified as transgender and 131 (50.4%) were self-identified as cisgender (nontransgender). Initial…

  5. Detection and Antibiotic Susceptibility Pattern of Biofilm Producing Gram Positive and Gram Negative Bacteria Isolated From a Tertiary Care Hospital of Pakistan

    Directory of Open Access Journals (Sweden)

    Iqbal, M.

    2011-01-01

    Full Text Available Microorganisms adhere to non-living material or living tissue, and form biofilms made up of extracellular polymers/slime. Biofilm-associated microorganisms behave differently from free-floating bacteria with respect to growth rates and ability to resist antimicrobial treatments and therefore pose a public health problem. The objective of this study is to detect the prevalence of biofilm producers among Gram positive and Gram negative bacteria isolated from clinical specimens, and to study their antimicrobial susceptibility pattern. The study was carried out from October 2009 to March 2010, at the Department of Microbiology, Army Medical College/ National University of Sciences and Technology (NUST, Rawalpindi, Pakistan. Clinical specimens were received from various wards of a tertiary care hospital. These were dealt by standard microbiological procedures. Gram positive and Gram negative bacteria isolated were subjected to biofilm detection by congo red agar method (CRA. Antimicrobial susceptibility testing of those isolates, which showed positive results (slime production, was done according to the Kirby-Bauer disc diffusion technique. A total of 150 isolates were tested for the production of biofilm/slime. Among them, 81 isolates showed positive results. From these 81, 51 were Gram positive and 30 were Gram negative. All the 81(54% slime producers showed reduced susceptibility to majority of antibiotics. Bacterial biofilms are an important virulence factor associated with chronic nosocomial infection. Detection of biofilm forming organisms can help in appropriate antibiotic choice.

  6. Ertapenem disk performance to predict Klebsiella pneumoniae carbapenemase produced by Gram-negative bacilli isolated in a São Paulo city public hospital.

    Science.gov (United States)

    Almeida, Lais Pinto de; Carvalho, Fabiana Puerro de; Marques, Alexandre Gimenes; Pereira, Andrea dos Santos; Bortoleto, Renata Puzzo; Martino, Marinês Dalla Valle

    2012-01-01

    To evaluate ertapenem disk performance to predict Klebsiella pneumonie carbapenemase production by Gram-negative bacilli. All Gram-negative bacilli isolated between January 2010 and June 2011 were tested by disk diffusion (Oxoid™) for sensitivity to ertapenem, meropenem and imipenem. Resistant or intermediate sensitivity strains (diameter Gram-negative bacilli were isolated in 708 (23.52%) of them. Zone of inhibition diameter for ertapenem disk was Gram-negative isolates. The PCR assay for blaKPC detected 40 Klebsiella pneumoniae carbapenemase-producing strains. No strains intermediate or resistant to meropenem and imipenem were sensitive to ertapenem. The ertapenem disk presented a positive predictive value of 36% to predict blaKPC and 89% specificity. The resistance of Gram-negative bacilli detected by disk diffusion against ertapenem does not predict Klebsiella pneumoniae carbapenemase production. Other mechanisms, such as production of other betalactamases and porin loss, may be implicated. The need to confirm the presence of the blaKPC is suggested. Therefore, ertapenem was a weak predictor for discriminating strains that produce Klebsiella pneumoniae carbapenemase.

  7. Isolates and antibiotic sensitivity of eighty culture-proven endophthalmitis cases from Istanbul.

    Science.gov (United States)

    Eser, Ilker; Kapran, Ziya; Altan, Tugrul; Ozel Karatas, Meltem; Aydin, Derya; Okaygun, Eda; Yilmaz, Omer Faruk

    2008-01-01

    To investigate the spectrum of organisms causing endophthalmitis and their sensitivity to commonly used antimicrobial agents. Medical records of 80 consecutive patients treated at Beyoglu Eye Hospital for endophthalmitis from January 2001 to April 2006 were reviewed. Specimens were obtained from either the vitreous (93%, 81/87) or anterior chamber (7%, 6/87) during pars plana vitrectomy or vitreous tap, and were inoculated into blood culture bottles. A Kirby-Bauer disk diffusion test was performed to determine antibiotic susceptibility. The outcome measures included isolates identified and antibiotic sensitivity of the specimens. Fifty-six of 87 (64.4%) isolates were Gram-positive organisms, 29 (33.3%) were Gram-negative organisms, and 2 (2.3%) were fungi. The most common organism group identified was coagulase-negative staphylococci in 26.4% (23/87). While vancomycin was active against all Gram-positive isolates tested (100%), ceftazidime had the highest susceptibility rate (100%) for Gram-negative organisms isolated. Although coagulase-negative micrococci predominated in this series, a high isolation rate for Gram-negative organisms was obtained. High susceptibility rates for ofloxacin make it an alternative to ceftazidime and vancomycin in both Gram-negative- and Gram-positive-derived endophthalmitis, respectively. Studies with larger series and additional antibiotics are needed to confirm these findings. (c) 2008 S. Karger AG, Basel

  8. Staphylococcal cassette chromosome mec typing and mecA sequencing in methicillin-resistant staphylococci from Algeria: a highly diversified element with new mutations in mecA.

    Science.gov (United States)

    Djoudi, Ferhat; Bonura, Celestino; Touati, Abdelaziz; Aléo, Aurora; Benallaoua, Said; Mammina, Caterina

    2016-11-01

    Genetic mechanisms of methicillin resistance are still relevant in staphylococci. The aims of this study are to assess the possible exchanges of staphylococcal cassette chromosome mec (SCCmec) among isolates of methicillin-resistant staphylococci (MRS) and to check for known or new mutations in mecA DNA. A total of 35 MRS non-repetitive isolates were recovered, including 20 Staphylococcushaemolyticus, 7 Staphylococcusaureus, 4 Staphylococcussciuri, 2 Staphylococcussaprophyticus and 1 isolate each of Staphylococcusxylosus and Staphylococcuslentus. Only 16 of the 35 strains were assigned to known SCCmec types: 7 SCCmec VII, 6 SCCmec IV and 3 SCCmec III, with possible horizontal transfer of the SCCmec VII from methicillin-resistant S. haemolyticus to methicillin-susceptible S. aureus. mecA gene sequencing in ten selected isolates allowed description of nine punctual mutations, seven of which were reported for the first time. The most frequent mutation was G246E, identified in isolates of methicillin-resistant S. aureus, S. sciuri, S. saprophyticus and S. lentus. These results emphasized the high degree of genetic diversity of SCCmec element in MRS and describe new missense mutations in mecA, which might be important in understanding the evolution of methicillin and new β-lactam resistance.

  9. Cultivation of vancomycin-resistant enterococci and methicillin-resistant staphylococci from input and output samples of German biogas plants.

    Science.gov (United States)

    Glaeser, Stefanie P; Sowinsky, Olivia; Brunner, Jana S; Dott, Wolfgang; Kämpfer, Peter

    2016-03-01

    Vancomycin-resistant enterococci (VRE) and methicillin-resistant staphylococci (MRS) were detected in two mesophilic German biogas plants (BGPs) using selective pre-enrichment methods combined with cultivation on CHROMagar media and antibiotic resistance gene screening. Genetic fingerprinting and 16S rRNA gene sequencing showed the presence of enterococci isolated by the VRE selective cultivation (67 isolates) in input and output samples of BGPs. In contrast, MRS (44 isolates) were detected in input, but in none of the output samples. Enterococcus isolates showed highest 16S rRNA gene sequence similarity (>99.8%) to E. lemanii, E. casseliflavus/E. gallinarium or E. devriesei/E. pseudoavium/E. viikkiensis and carried vanA, vanB and/or vanC1 genes. Enterococcus faecium and E. faecalis VRE were not detected, but isolates closely related to those species (>99.9% 16S rRNA gene sequence similarity) were detected by the MRS selective cultivation methods. Staphylococcus isolates shared highest 16S rRNA gene sequence similarity (>99.9%) with S. haemolyticus, S. lentus and S. sciuri and carried mecA genes. Methicillin-resistant S. aureus (MRSA) were not detected. In summary, manure from livestock husbandry contained both, VRE and MRS. VRE were also detected in output samples, indicating that enterococci with vancomycin resistance genes could be release into the environment by the application of BGP output material as biofertilizers.

  10. [In-vitro susceptibilites to levofloxacin and various antibacterial agents of 18,639 clinical isolates obtained from 77 centers in 2004].

    Science.gov (United States)

    Yamaguchi, Keizo; Ohno, Akira; Ishii, Yoshikazu; Tateda, Kazuhiro; Iwata, Morihiro; Kanda, Makoto; Tsujio, Yoshiko; Kimoto, Hiroya; Kaimori, Mitsuomi; Nakamura, Toshihiko; Kawamura, Chizuko; Nishimura, Masaharu; Akizawa, Koji; Katayama, Yosei; Matsuda, Keiko; Hayashi, Tasuku; Yasujima, Minoru; Kasai, Takeshi; Kimura, Masahiko; Tominaga, Makoto; Miki, Makoto; Nakanowatari, Susumu; Nakagawa, Takuo; Kaku, Mitsuo; Kanemitsu, Keiji; Kunishima, Hiroyuki; Toyoshima, Shunkoh; Sakurai, Masanori; Shiotani, Joji; Sugita, Akihiro; Ito, Tatsumi; Okada, Jun; Suwabe, Akira; Yamahata, Kumiko; Yoneyama, Akiko; Kumasaka, Kazunari; Yamane, Nobuo; Koike, Kazuhiko; Ieiri, Tamio; Kominami, Hidenori; Yamada, Toshiyuki; Oguri, Toyoko; Itoh, Kouichi; Watanabe, Kiyoaki; Kobayashi, Yoshio; Ohtake, Teruko; Uchida, Takashi; Totsuka, Kyoichi; Murakami, Masami; Yomoda, Sachie; Takahashi, Ayako; Okamoto, Hideyuki; Inuzuka, Kazuhisa; Yamazaki, Kenichiro; Gonda, Hideo; Yamashita, Takanori; Yamaguchi, Ikuo; Okada, Motoi; Ikari, Hiromi; Kurosawa, Naomi; Fujimoto, Yoshinori; Ishigo, Shiomi; Asano, Yuko; Mikio, Mori; Kano, Ichino; Nagano, Eiko; Kageyama, Fumio; Shaku, Etsuko; Kanno, Harushige; Aihara, Masanori; Gemma, Hitoshi; Uemura, Keiichi; Miyajima, Eiji; Maesaki, Shighefumi; Hashikita, Giichi; Horii, Toshinobu; Sumitomo, Midori; Yoshimura, Hitoshi; Hiraoka, Minoru; Wada, Hideo; Yuzuki, Yosuke; Ikeda, Norio; Baba, Hisashi; Soma, Masayuki; Yamamoto, Tsuyoshi; Ichiyama, Satoshi; Kinosita, Syohiro; Kawano, Seiji; Fujita, Shinichi; Kageoka, Takeshi; Hongo, Toshiharu; Okabe, Hidetoshi; Tatewaki, Kenichi; Moro, Kunihiko; Oka, Mikio; Niki, Yoshihito; Yoshida, Haruyoshi; Yamashita, Masanobu; Kusano, Nobuchika; Mihara, Eiichiro; Nose, Motoko; Fushiwaki, Takeshi; Kuwabara, Masao; Fujiue, Yoshihiro; Shimuzu, Akira; Takubo, Takayuki; Kusakabe, Tadashi; Hinoda, Yuji; Tanaka, Nobuaki; Takahashi, Hakuo; Heijyou, Hitoshi; Okazaki, Toshiro; Asai, Koji; Kawahara, Kunimitsu; Masuda, Junichi; Sano, Reiko; Taminato, Tomohiko; Negayama, Kiyoshi; Matsuo, Syuji; Komatsu, Masaru; Sugiura, Tetsuro; Murase, Mitsuharu; Hiramatsu, Kazufumi; Yamane, Nobuhisa; Nakasone, Isao; Hirakata, Yoichi; Kohno, Shigeru; Aizawa, Hisamichi; Honda, Junichi; Hamazaki, Naotaka; Okayama, Akihiko; Ono, Junko; Aoki, Yosuke; Okada, Kaoru; Miyanohara, Hiroaki

    2006-12-01

    A total of 18,639 clinical isolates in 19 species collected from 77 centers during 2004 in Japan were tested for their susceptibility to fluoroquinolones (FQs) and other selected antibiotics. The common respiratory pathogens, Streptococcus pyogenes, Streptococcus pneumoniae, Moraxella catarrhalis and Haemophilus influenzae showed a high susceptible rate against FQs. The isolation rate of beta lactamase non-producing ampicillin-resistant H. influenzae was approximately three times as large as those of western countries. Most strains of Enterobacteriaceae were also susceptible to FQs. The resistance rate of Escherichia coli against FQs has however been rapidly increasing so far as we surveyed since 1994. The FQs-resistant rate in methicillin-resistant Staphylococcus aureus (MRSA) showed approximately 90% except for 36%. of sitafloxacin while FQs-resistant rate in methicillin-susceptible S. aureus (MSSA) was around 5%. The FQs-resistant rate of methicillin-resistant coagulase negative Staphylococci (MRCNS) was also higher than that of methicillin-susceptible coagulase negative Staphylococci (MSCNS), however, it was lower than that of MRSA. In Pseudomonas aeruginosa clinical isolates, 32-34% from UTI and 15-19% of from RTI was resistant to FQs. Acinetobacter spp. showed a high susceptibility to FQs. Although FQs-resistant Neisseria gonorrhoeae have not been increased in western countries, it is remarkably high in Japan. In this survey, isolates of approximately 85% was resistant to FQs.

  11. Evaluation of cefoxitin disk diffusion breakpoint for detection of methicillin resistance in Staphylococcus pseudintermedius isolates from dogs.

    Science.gov (United States)

    Bemis, David A; Jones, Rebekah D; Videla, Ricardo; Kania, Stephen A

    2012-09-01

    Cefoxitin disk diffusion susceptibility testing is a recommended screening method for the detection of methicillin resistance in human isolates of Staphylococcus aureus and coagulase-negative staphylococci. A retrospective analysis of 1,146 clinical isolates of Staphylococcus pseudintermedius from dogs was conducted to determine if screening by the cefoxitin disk method can be similarly useful with S. pseudintermedius. The distribution of cefoxitin growth inhibition zone diameters within this collection was bimodal and correlated well with the results of methicillin resistance gene (mecA) detection by polymerase chain reaction. Of the isolates, 5% had discordant results and, when retested, 84% of these were in agreement. While a greater diversity of isolates and interlaboratory comparisons must be tested, the current study suggests that an epidemiological breakpoint (of approximately ≤ 30 mm = resistant; ≥ 31 = susceptible) can be established to predict methicillin resistance in S. pseudintermedius.

  12. PSM-Mec-A Virulence Determinant that Connects Transcriptional Regulation, Virulence, and Antibiotic Resistance in Staphylococci.

    Science.gov (United States)

    Qin, Li; McCausland, Joshua W; Cheung, Gordon Y C; Otto, Michael

    2016-01-01

    PSM-mec is a secreted virulence factor that belongs to the phenol-soluble modulin (PSM) family of amphipathic, alpha-helical peptide toxins produced by Staphylococcus species. All known PSMs are core genome-encoded with the exception of PSM-mec, whose gene is found in specific sub-types of SCCmec methicillin resistance mobile genetic elements present in methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci. In addition to the cytolytic translational product, PSM-mec, the psm-mec locus encodes a regulatory RNA. In S. aureus, the psm-mec locus influences cytolytic capacity, methicillin resistance, biofilm formation, cell spreading, and the expression of other virulence factors, such as other PSMs, which results in a significant impact on immune evasion and disease. However, these effects are highly strain-dependent, which is possibly due to differences in PSM-mec peptide vs. psm-mec RNA-controlled effects. Here, we summarize the functional properties of PSM-mec and the psm-mec RNA molecule and their roles in staphylococcal pathogenesis and physiology.

  13. Antibacterial susceptibility profiles of subclinical mastitis pathogens isolated in Batna and Setif Governorates (East of Algeria

    Directory of Open Access Journals (Sweden)

    Mamache Bakir

    Full Text Available Sub-clinical mastitis is a main pathology of dairy husbandry because it is not clinically recognized by the owners and the veterinarians. For this reason, its economic loss is usually underestimated in milk production. This study has been undertaken in order to evaluate the epidemiologic situation of sub-clinicalmastitis in Batna and Setif governorates (East of Algeria. For this purpose, a detailed bacteriological study of all bacterial strains isolated from sub-clinical mastitis followed by a study of their antibacterial susceptibility profiles has been undertaken. 89 bacterial strains distributed as follows were studied: 27 strains of staphylococci among which 23 were coagulase-negative staphylococci (CNS that are generally incriminated in sub-clinical mastitis. 39 strains of streptococci among which 10 were Lactococcus lactis ssp lactis strains. 23 strains of enterobacteria represented mainly by Escherichia coli (E.coli. All these bacterial strains were isolated from cow milk of 3 different farms. The antibacterial susceptibility profiles have revealed a susceptibility of the isolated strains to a large number of antibiotics mainly to the Neomycin, the Cephalexin and the Spiramycin. [Vet. World 2011; 4(12.000: 537-541

  14. Isolation and identification of bacterial causes of clinical mastitis in cattle in Sulaimania region

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    S. A. Hussein

    2008-01-01

    Full Text Available A total of 51 cases of bovine clinical mastitis in Sulaimani district were investigated for their bacteriological causative agents; 76 milk samples were cultured on primary and selective media and the isolated bacteria were tested for their susceptibility to antimicrobial agents used in commercial intramammary infusion products. Eighty two bacterial isolates were obtained and further identified using biochemical tests. Escherichia coli was the most common bacteria followed by Staphylococcus aureus, Streptococcus agalactia and coagulase–negative staphylococci. Two other bacterial species (Pseudomonas aeruginosa and Streptococcucs uberis were also isolated but in a lower proportion. Antibacterial susceptibility testing showed that the use of florfenicol, cephalexin and gentamicin may be useful for the treatment of clinical mastitis cases in cows.

  15. Theoretical Design and Characteristics Analysis of a Quasi-Zero Stiffness Isolator Using a Disk Spring as Negative Stiffness Element

    Directory of Open Access Journals (Sweden)

    Lingshuai Meng

    2015-01-01

    Full Text Available This paper presents a novel quasi-zero stiffness (QZS isolator designed by combining a disk spring with a vertical linear spring. The static characteristics of the disk spring and the QZS isolator are investigated. The optimal combination of the configurative parameters is derived to achieve a wide displacement range around the equilibrium position in which the stiffness has a low value and changes slightly. By considering the overloaded or underloaded conditions, the dynamic equations are established for both force and displacement excitations. The frequency response curves (FRCs are obtained by using the harmonic balance method (HBM and confirmed by the numerical simulation. The stability of the steady-state solution is analyzed by applying Floquet theory. The force, absolute displacement, and acceleration transmissibility are defined to evaluate the isolation performance. Effects of the offset displacement, excitation amplitude, and damping ratio on the QZS isolator and the equivalent system (ELS are studied. The results demonstrate that the QZS isolator for overloaded or underloaded can exhibit different stiffness characteristics with changing excitation amplitude. If loaded with an appropriate mass, excited by not too large amplitude, and owned a larger damper, the QZS isolator can possess better isolation performance than its ELS in low frequency range.

  16. Virulence factors and ability of staphylococci from bovine milk and the cowshed environment to biofilm formation.

    Science.gov (United States)

    Kot, B; Binek, T; Piechota, M; Wolska, K M; Zdunek, E; Platkowska, K

    2013-01-01

    The aim of this study was to examine virulence factors and the ability of S. aureus and CNS species isolated from milk of cows with mastitis to form biofilm, and to compare them with virulence factors of staphylococci from milk of cows without mastitis and cowshed environment. Most of S. aureus strains from cows with mastitis showed haemolytic activity (93.9%), among them 72.7% and 21.2% produced alpha- and beta-haemolysin, respectively. S. aureus from cows with mastitis symptoms produced proteases (above 48%) and esterase (42.4%). The highly significant relationship between the number of S. xylosus strains producing haemolysins (62%) and the origin of these strains from milk of cows with mastitis was observed. The ability to produce proteases was significantly associated with S. sciuri from milk of cows with mastitis. The ability of biofilm formation by staphylococcal strains from milk of cows with mastitis was greater than in strains from milk of cows without mastitis and the difference was significant (p < or = 0.05). The highest percentage of strains from milk of cows with mastitis were weak biofilm formers (48.6%), while 40% and 11.4% of strains were moderate and strong biofilm producers, respectively. S. xylosus showed the highest ability to form biofilm, while the lowest ability to form biofilm was observed in S. aureus and S. epidermidis. In conclusion, production of exotoxins and enzymes, and ability of biofilm formation shown by many CNS isolated from milk of cows with mastitis symptoms indicates that these features are important in pathogenesis of this disease.

  17. Multiresistant Bacteria Isolated from Chicken Meat in Austria

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    Gernot Zarfel

    2014-12-01

    Full Text Available Multidrug resistant bacteria (MDR bacteria, such as extended spectrum beta-lactamase (ESBL Enterobacteriaceae, methicillin resistant Staphylococcus aureus (MRSA, and vancomycin-resistant Enterococci (VRE, pose a challenge to the human health care system. In recent years, these MDR bacteria have been detected increasingly outside the hospital environment. Also the contamination of food with MDR bacteria, particularly of meat and meat products, is a concern. The aim of the study was to evaluate the occurrence of MDR bacteria in chicken meat on the Austrian market. For this study, 50 chicken meat samples were analysed. All samples originated from chickens slaughtered in Austrian slaughterhouses and were marked as produced in Austria. Samples were analysed for the presence of ESBL Enterobacteriaceae, methicillin resistant Staphylococci and VRE. Resistance genes of the isolated bacteria were characterised by PCR and sequencing. In the present study 26 ESBL producing E. coli, five mecA gene harbouring Staphylococci (but no MRSA, and four VRE were detected in chicken meat samples of Austrian origin. In 24 (48% of the samples no ESBL Enterobacteriaceae, MRSA, methicillin resistant coagulase negative Staphylococcus (MRCNS or VRE could be detected. None of the samples contained all three types of investigated multiresistant bacteria. In concordance to previous studies, CTX-M-1 and SHV-12 were the dominant ESBL genes.

  18. Comparison phenotypic and genotypic identification of Staphylococcus species isolated from bovine mastitis

    Directory of Open Access Journals (Sweden)

    Felipe Freitas Guimarães

    Full Text Available ABSTRACT: In addition to Staphylococcus aureus nowadays other coagulase-positive staphylococci (CoPS and coagulase-negative staphylococci (CoNS, earlier considered of minor importance, are now accepted as relevant pathogens for humans and animals. The involvement of these microorganisms in bovine mastitis etiology and the possibility their transmission through milk to humans justify the requirement of developing reliable methods for identification of the most frequent species among them. The purpose of this study was to compare the phenotypic techniques with the genotypic method carried out by sequencing of the rpoB gene in identification of several species of the genus Staphylococcus isolated from bovine mastitis. A total of 300 staphylococci isolates of bovine mastitis cases from several Brazilian dairy herds were studied by phenotypic and genotypic techniques, respectively: 150 CoPS and 150 CoNS strains. A total of 18 CoNS different species and 4 CoPS species were identified. Among the CoNS the following species were recognized: 48 (32% Staphylococcus warneri, 22(15% S. epidermidis, 20(13% S. hyicus, 10(7% S. xylosus, 7(5% S. haemolyticus, 6(4% S. simulans, 6(4% S. schleiferi subsp schleiferi, 6(4% S. hominis, 5(3% S. pasteuri, 4(2.7% S. cohnii, 3(2% S. saprophyticus subsp. saprophyticus 3(2% S. chromogenes 3(2% S. sciuri, 2(1% S. saccharolyticus, 2(1% S. lugdunensi, 1(0,7% S. auricularis, 1(70% S. saprophyticus subsp. bovis, 1(0.7% S. capitis. And among the 150 CoPS were identified respectively: 105 (70% S. aureus, 21(14%, S. hyicus, 19(13% S. intermedius e 5(3% S. schleiferi subsp coagulans. Considering the 150 CoNS isolates, the identifications performed by phenotypic and genotypic tests presented 96.7% of concordance, kappa coefficient of agreement = 0.933, SE (standard error of kappa=0.021 (95% confidence interval: 0.893 to 0.974, Pearson’s correlation coefficient (r = 0.9977, (confidence interval 95%: 0.9938 a 0.9992 and in relation

  19. Antibiotic susceptibility pattern in urinary isolates of gram negative bacilli with special reference to AmpC β-lactamase in a tertiary care hospital

    Directory of Open Access Journals (Sweden)

    Patel Mitesh

    2010-01-01

    Full Text Available Introduction: Resistance to higher antimicrobial agent is commonly seen in gram negative bacilli. This issue is a challenging problem to the medical practitioners in addition to it is financial impact on the health care system. Objectives: To document the prevalence of multi drug resistant gram negative bacilli isolated from urine of patients attending the Urology Department of Tertiary care Hospital of western India in year 2008. Results: Out of total 328 isolates, 118 (35.98% E.coli, 72 (21.95 % Klebsiella, 64 (19.51% Pseudomonas aeruginosa, 30 (9.15% Acinetobacter, 18 (5.49% Proteus vulgaris, 18 (5.49% Proteus mirabilis, 6 (1.83% Providencia rettgerii, 2 (0.61% Citrobacter freundii. Out of these isolates, 228 (69.51% were b-lactamase positive, while 100 (30.51% were β-lactamase negative. Out of 228 β-lactamase positive, 104 (45.61% were AmpC β-lactamase positive. Conclusions: Stringent protocol such as Antibiotic policy and Hospital infection control program are mandatory to curb these microbes in a tertiary care hospital.

  20. Antibiotic susceptibility pattern in urinary isolates of gram negative bacilli with special reference to AmpC β-lactamase in a tertiary care hospital

    Science.gov (United States)

    Patel, Mitesh H.; Trivedi, Grishma R.; Patel, Sachin M.; Vegad, Mahendra M.

    2010-01-01

    Introduction: Resistance to higher antimicrobial agent is commonly seen in gram negative bacilli. This issue is a challenging problem to the medical practitioners in addition to it is financial impact on the health care system. Objectives: To document the prevalence of multi drug resistant gram negative bacilli isolated from urine of patients attending the Urology Department of Tertiary care Hospital of western India in year 2008. Results: Out of total 328 isolates, 118 (35.98%) E.coli, 72 (21.95 %) Klebsiella, 64 (19.51%) Pseudomonas aeruginosa, 30 (9.15%) Acinetobacter, 18 (5.49%) Proteus vulgaris, 18 (5.49%) Proteus mirabilis, 6 (1.83%) Providencia rettgerii, 2 (0.61%) Citrobacter freundii. Out of these isolates, 228 (69.51%) were β-lactamase positive, while 100 (30.51%) were β-lactamase negative. Out of 228 β-lactamase positive, 104 (45.61%) were AmpC β-lactamase positive. Conclusions: Stringent protocol such as Antibiotic policy and Hospital infection control program are mandatory to curb these microbes in a tertiary care hospital. PMID:20842250

  1. The distribution and antimicrobial resistance analysis of pathogens isolated from blood specimens of neonates%新生儿血培养病原菌分布及耐药性分析

    Institute of Scientific and Technical Information of China (English)

    李响新; 胡型忠; 陈秀平; 项领

    2011-01-01

    目的:探讨新生儿血培养病原菌分布及耐药性,为临床合理使用抗菌药提供依据.方法:对我院新生儿科2009年6月-2011年6月的血培养细菌分布及耐药性进行回顾性分析.结果:1868例新生儿血培养标本共检出细菌210株,阳性率为11.2%.细菌以凝固酶阴性葡萄球菌为主(160株,占76.2%),革兰阴性杆菌仅23株占11.0%,以大肠埃希菌和肺炎克雷伯菌为主.葡萄球菌属对青霉素的敏感率最低为7.7%,苯唑西林为31.4%;粪肠球菌对万古霉素、替考拉宁100.0%敏感;革兰阴性杆菌对阿莫西林的敏感率<30.0%,抗菌活性较好的药物是哌拉西林/他唑巴坦、亚胺培南和美罗培南.结论:新生儿血培养病原菌以凝固酶阴性葡萄球菌为主,其耐药性强,因此,加强血培养的检测和药敏试验对指导临床医师合理使用抗菌药非常有必要.%Objective: To investigate distribution and antimicrobial resistance of pathogens causing neonatal septicemia, and to provide references for clinical treatment. Methods: The data of distribution and antimicrobial resistance of pathogens collected from blood specimens of neonatal sepsis patients from June 2009 to June 2011 in our hospital were analyzed retrospectively. Re-S;lltS: 210 pathogenic strains were isolated from 1868 blood specimens, and the positive rate was 11. 2%. The strains of coagu-lase - negative staphylococci ( CNS) were the most frequent detected isolates, accounting for 76. 2% . Only 23 gram - negative strains were isolated, accounting for 11.0% , while escherichia coli and Klebsiella pneumoniae ranked in top two. Staphylococci showed the lowest susceptibility rate against penicillin was 7. 7% while oxacillin was 31. 4%. All the Enterococcus strains were susceptible to teicoplanin and vancomycin. The susceptibility rate of Gram - negative bacteria to amoxicillin was lower than 30% , whereas piperacillin/tazobactam, imipenem and meropenem maintained

  2. Biofilm Formation and the Presence of the Intercellular Adhesion Locus ica among Staphylococci from Food and Food Processing Environments

    OpenAIRE

    Møretrø, Trond; Hermansen, Lene; Holck, Askild L.; Sidhu, Maan S.; Rudi, Knut; Langsrud, Solveig

    2003-01-01

    In clinical staphylococci, the presence of the ica genes and biofilm formation are considered important for virulence. Biofilm formation may also be of importance for survival and virulence in food-related staphylococci. In the present work, staphylococci from the food industry were found to differ greatly in their abilities to form biofilms on polystyrene. A total of 7 and 21 of 144 food-related strains were found to be strong and weak biofilm formers, respectively. Glucose and sodium chlori...

  3. Genetic characterisation of carbapenem-resistant Gram-negative bacteria isolated from the University Hospital Mohamed Boudiaf in Ouargla, southern Algeria.

    Science.gov (United States)

    Yagoubat, Mounira; Ould El-Hadj-Khelil, Aminata; Malki, Ali; Bakour, Sofiane; Touati, Abdelaziz; Rolain, Jean-Marc

    2017-03-01

    The aim of this study was to investigate the molecular epidemiology and the genetic support of carbapenem-resistant Enterobacteriaceae and Acinetobacter spp. isolates collected in the University Hospital of Ouargla, southern Algeria. A total of 99 Gram-negative bacteria (GNB) were collected from stool samples of colonised patients and from inanimate surfaces in the hospital environment between December 2014 and August 2015. Selected Enterobacteriaceae and Acinetobacter spp. isolates with reduced susceptibility to carbapenems were subjected to phenotypic study, including antibiotic susceptibility testing according to CA-SFM-EUCAST 2015 guidelines and modified Carba NP test. Genes encoding carbapenemases, extended-spectrum β-lactamases (ESBLs) and AmpC β-lactamases were screened by PCR and sequencing. Clonal relatedness was determined by multilocus sequencing typing (MLST). Of the 99 GNB isolates, 10 (10.1%) showed reduced susceptibility to carbapenems were studied further, including 7 Acinetobacter baumannii, 1 Acinetobacter nosocomialis, 1 Escherichia coli and 1 Klebsiella pneumoniae. PCR and sequencing showed that four A. baumannii isolates and the single A. nosocomialis isolate harboured blaNDM-1. In addition, blaOXA-23 was observed in three A. baumannii isolates, and blaOXA-48 was detected in the two Enterobacteriaceae isolates. MLST assigned the K. pneumoniae to ST999 and the E. coli to ST38. The seven A. baumannii isolates belonged to ST85 and ST2. This study describes the epidemiology of carbapenemases produced by Enterobacteriaceae and Acinetobacter spp. in southern Algeria and reports the first description of metallo-β-lactamase NDM-1-producing A. nosocomialis in Algeria. Copyright © 2016 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.

  4. Caracterização fenotípica da resistência a antimicrobianos e detecção do gene mecA em Staphylococcus spp. coagulase-negativos isolados de amostras animais e humanas Phenotypic characterization of antimicrobial resistance and detection of the mecA gene in coagulase-negative Staphylococcus spp. isolates from animal and human samples

    Directory of Open Access Journals (Sweden)

    Lidiane de Castro Soares

    2008-08-01

    Full Text Available Os estafilococos coagulase-negativos (ECN fazem parte da microbiota normal da pele e, apesar de terem sido considerados saprófitas por muito tempo, o seu significado clínico como agente etiológico tem aumentado com o passar dos anos. Neste estudo, foram obtidos 72 isolados de ECN a partir de amostras do conduto auditivo de cães, de mastite bovina e de infecções humanas. Staphylococcus xylosus foi o microrganismo mais isolado, nas amostras animais, e S. cohnii subsp. cohnii em humanos. Os isolados foram avaliados de modo a traçar o perfil fenotípico de sua resistência aos antimicrobianos mais indicados no tratamento de infecções estafilocócicas. Foi detectado um elevado nível de resistência à penicilina e ampicilina. A gentamicina, a vancomicina e a associação ampicilina+sulbactam foram eficientes frente aos isolados testados. A resistência à oxacilina foi avaliada por meio dos testes de difusão em disco modificada, ágar screen, microdiluição em caldo e diluição em ágar para constatar, se à semelhança do que ocorre com os estafilococos coagulase-positivo, esta pode ser mediada pelo gene mecA e apresentada de forma heterogênea. A presença do gene mecA foi determinada pelo método da Reação em Cadeia de Polimerase (PCR, sendo 5,6% dos isolados mecA positivos.Coagulase-negative staphylococci (SCN make part of the normal microbiota skin and although they have been considered saprophytics for years, nowadays their clinical significance as an etiologic agent has increased. In this study, 72 SCN isolates obtained from external ear canals of dogs, bovine mastitis and human nosocomial infections were evaluated. Staphylococcus xylosus was the most prevalent microorganism in animal samples and S. cohnii subsp. cohnii in human samples. SCN isolates were evaluated in order to establish a phenotypical resistance pattern towards the most indicated antibiotics for staphyloccocal infections. A high level of resistance to penicillin

  5. Prevalencia y perfil de resistencia a betalactámicos en estafilococos de perros y gatos Prevalence and resistance profile to beta-lactam antibiotics in staphylococci from dogs and cats

    Directory of Open Access Journals (Sweden)

    G Denamiel

    2009-12-01

    Full Text Available El objetivo de este trabajo fue conocer la prevalencia de especies y perfil de resistencia a betalactámicos en estafilococos aislados de infecciones en caninos y felinos. En perros, de 207 aislamientos, el porcentaje de las especies identificadas fue: S. intermedius 29,5%, S. aureus y S. simulans 20,8%, S. saprophyticus 12,6%, S. chromogenes 10,6%, S. haemolyticus 3,9% y S. epidermidis 1,9%. De 63 aislamientos en gatos: S. simulans 33,3%, S. intermedius y S. aureus 15,4%, S. saprophyticus 11,1%, S. haemolyticus 6,34%, S. chromogenes y S. epidermidis 3,5%. En perros, el porcentaje de resistencia a penicilina fue: S. intermedius 47,5%, S. aureus 34,8% y S. saprophyticus 7,7%. Los aislamientos que expresaron meticilina resistencia correspondieron: S. intermedius 9,8% y S. saprophyticus 3,8%. En gatos los estafilococos resistentes a penicilina fueron S. saprophyticus 42,8%, S. aureus y S. intermedius 15,4% y S. simulans 9,5%. No se hallaron especies de estafilococos meticilino resistentes.The purpose of this study was to characterize the prevalence and resistance profile to betalactam antibiotics of staphylococci isolated from canine and feline infections. Staphylococci were isolated from 207 dog samples. The species frequency for identified staphylococci were: S. intermedius 29.5%, S. aureus and S. simulans 20.8%, S. saprophyticus 12.6%, S. chromogenes 10.6%, S. haemolyticus 3.9% and S. epidermidis 1.9%. Sixty three staphylococci isolates were from cats: S. simulans 33.3%, S. intermedius and S. aureus 15.4%, S. saprophyticus 11.1%, S. haemolyticus 6.34%, S. chromogenes and S. epidermidis 3.5%. Resistance to penicillin in samples from dogs were: S. intermedius 47.5%, S. aureus 34.8% and S. saprophyticus 7.7%. Methicillin-resistance was evidenced in S. intermedius 9.8% and S. saprophyticus 3.8%. In samples from cats, penicillin-resistant staphylococci were: S. saprophyticus 42.8%, S. aureus and S. intermedius 15.4% and S. simulans 9

  6. Antimicrobial resistance among Gram-negative bacilli isolated from Latin America: results from SENTRY Antimicrobial Surveillance Program (Latin America, 2008-2010).

    Science.gov (United States)

    Gales, Ana C; Castanheira, Mariana; Jones, Ronald N; Sader, Hélio S

    2012-08-01

    This study updates the frequency and resistance rates of Gram-negative bacilli isolated from Latin American medical centers enrolled in the SENTRY Antimicrobial Surveillance Program. A total of 12,811 bacterial organisms, including 5704 Gram-negative bacilli (44.5%), were consecutively collected (1 per patient) between January 2008 and December 2010 from 10 Latin American medical centers located in Argentina, Brazil, Chile, and Mexico. Antimicrobial susceptibility testing was performed and interpreted by the Clinical and Laboratory Standards Institute broth microdilution method at a central laboratory. All Gram-negative organisms with reduced susceptibility to imipenem or meropenem (MIC, ≥ 2 μg/mL) were screened for carbapenemase production by the modified Hodge test and by polymerase chain reaction. ESBL rates were 18.1%, 12.8%, 23.8%, and 48.4% among Escherichia coli and 60.4%, 49.9%, 59.2%, and 33.3% among Klebsiella spp. from Argentina, Brazil, Chile, and Mexico, respectively. Meropenem-nonsusceptible Klebsiella spp. rate was highest in Brazil (11.1%), followed by Argentina (8.2%), Chile (5.0%), and Mexico (0.8%). Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae was not detected in 2008, but emerged in 2009 (10 strains) and increased significantly in 2010 (44; P 77% overall coverage against the 5 most frequently isolated Gram-negative bacilli from Latin American Medical centers participating in the SENTRY Program. Copyright © 2012 Elsevier Inc. All rights reserved.

  7. Prevalence of enterotoxin genes and antimicrobial resistance of coagulase-positive staphylococci recovered from raw cow milk.

    Science.gov (United States)

    Rola, J G; Korpysa-Dzirba, W; Czubkowska, A; Osek, J

    2015-07-01

    Raw milk may be contaminated by enterotoxigenic coagulase-positive staphylococci (CPS). Several of these microorganisms show antimicrobial resistance, which poses a potential risk for consumers. The aim of this study was to determine the occurrence of enterotoxin genes and antimicrobial resistance of CPS isolated from cow milk. A total of 115 samples were analyzed for the presence of CPS according to the International Organization for Standardization standard (ISO 6888-2). The genes were identified using 2multiplex PCR assays. Resistance of the isolates to 10 antimicrobials was determined using the minimum inhibitory concentration method. Overall, 71 samples (62%) were contaminated with CPS and 69 isolates were further analyzed. Among them, 20 (29%) strains harbored the enterotoxin genes. The most commonly detected staphylococcal enterotoxin markers were sed, sej, and ser, whereas none of the analyzed isolates possessed the seb and see genes. Almost one-half of the tested strains (43%) were resistant to one or more antimicrobial agents. Resistance to penicillin was the most common, followed by sulfamethoxazole and chloramphenicol. On the other hand, all strains were susceptible to ciprofloxacin, erythromycin, gentamicin, cefoxitin, and streptomycin. None of the strains was positive for the mecA and mecC (methicillin-resistant Staphylococcus aureus) genes. These results indicate that enterotoxigenic and antimicrobial-resistant CPS strains are present in raw milk, which may be a potential risk for public health.

  8. Boronic acid disk diffusion for the phenotypic detection of polymerase chain reaction-confirmed, carbapenem-resistant, gram-negative bacilli isolates.

    Science.gov (United States)

    Elsherif, Rasha; Ismail, Dalia; Elawady, Sanaa; Jastaniah, Samyah; Al-Masaudi, Saad; Harakeh, Steve; Karrouf, Gamal

    2016-07-01

    The Middle East is regarded as a secondary reservoir for OXA-48 and New Delhi metallo-β-lactamase (NDM) carbapenemases. One of the main challenges in clinical microbiology diagnostics is the detection of carbapenemases. For this reason simple screening methods have been sought to detect carbapenemase producers to determine appropriate therapeutic measures and implement infection control interventions. The present study aimed to evaluate the efficacy of the modified Hodge test (MHT) and a boronic acid-based combined disk test using carbapenems as substrates for the phenotypic determination of OXA-48 and NDM type carbapenemases in 45 epidemiologically unrelated carbapenem-resistant clinical isolates of Klebsiella pneumoniae (13 isolates), Acinetobacter baumanii (20 isolates), and Pseudomonas aeruginosa (12 isolates). Boronic acid disk test using meropenem as substrate and 600 µg of 3- aminophenylboronic acid (APB) was the most sensitive method (83.33 %) for detection of OXA-48, while the most specific method was MHT (100 %). As regards NDM carbapenemase, boronic acid disk tests using imipenem and 600 µg of APB per disk, and meropenem with 300 or 600 µg of APB were the most  sensitive methods (87.50 %), while the most specific method was the MHT (100 %). The results of the present study indicate that phenotypic screening with the MHT and the boronic acid disk test may be used to detect OXA-48 and NDM carbapenemases in Gram-negative bacilli clinical isolates, and that these tests can be easily applied in tertiary care settings with minimal infrastructure.

  9. Postoperative mediastinitis due to Finegoldia magna with negative blood cultures.

    Science.gov (United States)

    Kernéis, Solen; Matta, Matta; Hoï, Annie Buu; Podglajen, Isabelle; Gutmann, Laurent; Novara, Ana; Latremouille, Christian; Mainardi, Jean-Luc

    2009-12-01

    We report a case of Finegoldia magna (formerly known as Peptostreptococcus magnus) mediastinitis following coronary artery bypass in a 50-year-old patient. Even if staphylococci remain the main causative organism of postoperative mediastinitis, the responsibility of anaerobic bacteria must be considered in cases of fever and sternal drainage with negative blood cultures.

  10. Nonfermentative gram-negative microorganisms isolated from intensive care units and their resistance profiles in a training and research hospital

    Directory of Open Access Journals (Sweden)

    Fulya Bayındır Bilman

    2014-09-01

    Full Text Available Objective: The aim of this study was to determine the pathogenic agents, which have been isolated in ICUs and determine their antimicrobial susceptibility in the first two years of our hospital. Methods: A total 368 strains (194 Pseudomonas aeruginosa, and 174 Acinetobacter baumannii that were considered as an infectious agent and isolated from different clinical samples of hospitalized patients in intensive care units between January 2011 and December 2012 were included in the study. Conventional methods and automatized system were used for the identification and antibiotic susceptibility of the isolates with according to guidelines of Clinical and Laboratory Standarts Institute (CLSI. Imipenem and meropenem resistance has been confirmed by the E test method and interpreted according to CLSI. Results: Resistance rates of P.aeruginosa strains were as follows: Meropenem 35%, imipenem 36%, cefepim 21%, ceftazidim and ciprofloxacin 29%, gentamicin 33%, piperacilin/tazobactam 35%, amikacin 37%. Resistance rates of A.baumannii strains were as follows: Meropenem 85%, imipenem 87%, amikacin 22%, trimethoprim/sulfamethoxazole 25%, gentamicin 46%, piperacilin/tazobactam 54%, ampicilin/sulbactam 59%, cefepim 64%, ciprofloxacin 71%, ceftazidim 83%. Conclusion: Increased carbapenem resistance in our ICUs demonstrates importance of antimicrobial susceptibility tests. The antibiotic susceptibility tests should be surveyed continuously to avoid the spread of intensive care unit isolates carrying high level antibiotic resistance for better achievement of treatment regimens. J Clin Exp Invest 2014; 5 (3: 391-396

  11. Isolation and purification of enterocin E-760 with broad antimicrobial activity against gram-positive and gram-negative bacteria.

    Science.gov (United States)

    Line, J E; Svetoch, E A; Eruslanov, B V; Perelygin, V V; Mitsevich, E V; Mitsevich, I P; Levchuk, V P; Svetoch, O E; Seal, B S; Siragusa, G R; Stern, N J

    2008-03-01

    Strain NRRL B-30745, isolated from chicken ceca and identified as Enterococcus durans, Enterococcus faecium, or Enterococcus hirae, was initially identified as antagonistic to Campylobacter jejuni. The isolate produced a 5,362-Da bacteriocin (enterocin) that inhibits the growth of Salmonella enterica serovar Enteritidis, S. enterica serovar Choleraesuis, S. enterica serovar Typhimurium, S. enterica serovar Gallinarum, Escherichia coli O157:H7, Yersinia enterocolitica, Citrobacter freundii, Klebsiella pneumoniae, Shigella dysenteriae, Pseudomonas aeruginosa, Proteus mirabilis, Morganella morganii, Staphylococcus aureus, Staphylococcus epidermidis, Listeria monocytogenes, Campylobacter jejuni, and 20 other Campylobacter species isolates. The enterocin, E-760, was isolated and purified by cation-exchange and hydrophobic-interaction chromatographies. The proteinaceous nature of purified enterocin E-760 was demonstrated upon treatment with various proteolytic enzymes. Specifically, the antimicrobial peptide was found to be sensitive to beta-chymotrypsin, proteinase K, and papain, while it was resistant to lysozyme and lipase. The enterocin demonstrated thermostability by retaining activity after 5 min at 100 degrees C and was stable at pH values between 5.0 and 8.7. However, activity was lost below pH 3.0 and above pH 9.5. Administration of enterocin E-760-treated feed significantly (P strains of C. jejuni by more than 8 log(10) CFU. Enterocin E-760 also significantly (P < 0.05) reduced the colonization of naturally acquired Campylobacter species in market age broiler chickens when administered in treated feed 4 days prior to analysis.

  12. Comparison of in-house and commercial real time-PCR based carbapenemase gene detection methods in Enterobacteriaceae and non-fermenting gram-negative bacterial isolates.

    Science.gov (United States)

    Smiljanic, M; Kaase, M; Ahmad-Nejad, P; Ghebremedhin, B

    2017-07-10

    Carbapenemase-producing gram-negative bacteria are increasing globally and have been associated with outbreaks in hospital settings. Thus, the accurate detection of these bacteria in infections is mandatory for administering the adequate therapy and infection control measures. This study aimed to establish and evaluate a multiplex real-time PCR assay for the simultaneous detection of carbapenemase gene variants in gram-negative rods and to compare the performance with a commercial RT-PCR assay (Check-Direct CPE). 116 carbapenem-resistant Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter baumannii isolates were genotyped for carbapenemase genes by PCR and sequencing. The defined isolates were used for the validation of the in-house RT-PCR by use of designed primer pairs and probes. Among the carbapenem-resistant isolates the genes bla KPC, bla VIM, bla NDM or bla OXA were detected. Both RT-PCR assays detected all bla KPC, bla VIM and bla NDM in the isolates. The in-house RT-PCR detected 53 of 67 (79.0%) whereas the commercial assay detected only 29 (43.3%) of the OXA genes. The in-house sufficiently distinguished the most prevalent OXA types (23-like and 48-like) in the melting curve analysis and direct detection of the genes from positive blood culture vials. The Check-Direct CPE and the in-house RT-PCR assay detected the carbapenem resistance from solid culture isolates. Moreover, the in-house assay enabled the identification of carbapenemase genes directly from positive blood-culture vials. However, we observed insufficient detection of various OXA genes in both assays. Nevertheless, the in-house RT-PCR detected the majority of the OXA type genes in Enterobacteriaceae and A. baumannii.

  13. Antimicrobial susceptibility pattern of bacterial isolates from wound infection and their sensitivity to antibiotic agents at super specialty hospital, Amravati city, India

    Directory of Open Access Journals (Sweden)

    Hrishikesh Sawdekar

    2015-02-01

    Full Text Available Background: Wound infection is one of the health problems that is caused and aggravated by the invasion of pathogenic organisms. Information on local pathogens and sensitivity to antimicrobial agent is crucial for successful treatment of wounds. So the present study was conducted to determine antimicrobial susceptibility pattern of bacterial isolates from wound infection and their sensitivity to antimicrobial agents. Methods: A retrospective study was conducted among patients with wound infection in Suyash super speciality hospital, from January 2012 to December 2013. Wound swab was collected using sterile cotton swabs and processed for bacterial isolation and susceptibility testing to Systemic antimicrobial agents. Results: In this study 78 bacterial isolates were recovered from 258 specimens showing an isolation rate of 31.2%. The predominant bacteria isolated from wounds were gram positive staphylococci 36 (46.2%, followed by gram negative streptococci 18 (23.1% gram negative pseudomonas 12 (15.4 % and gram negative proteus 8 (10.4%. The gram positive and gram negative bacteria constituted 68 (87.2% and 10 (12.8% of bacterial isolates; respectively. Conclusion: In the present study most of the pathogens isolated from wound isolates showed high rate of resistance to most commonly used newer antibiotics used to treat bacterial infections. Therefore, rational use of antibiotics should be practiced. [Int J Res Med Sci 2015; 3(2.000: 433-439

  14. Nosocomial Infections: Multicenter surveillance of antimicrobial resistance profile of Staphylococcus aureus and Gram negative rods isolated from blood and other sterile body fluids in Iran

    Directory of Open Access Journals (Sweden)

    Bahman Poorabbas

    2015-11-01

    Full Text Available Background and Objective: Antibiotic resistance is increasing, especially in healthcare-associated infections causing significant public health concerns worldwide. National information is required to make appropriate policies, update list of essential drugs for treatment, and evaluate the effects of intervention strategies. A nationwide surveillance of antimicrobial resistant bacteria in nosocomial infections was established in Iran in 2008, so that the data obtained through the surveillance would enable us to construct a database.Materials and Methods: Seven major teaching hospitals in Shiraz, Tabriz, Sari, Mashhad, Sanandaj, Ahwaz and Isfahan participated in this study. A total of 858 strains isolated from blood and other sterile body fluids were tested. Identification at the species level was performed with conventional biochemical methods and the API system. Susceptibility tests were done using disk diffusion method. The methicillin-resistance in S. aureus (MRSA was determined by the oxacillin agar screen plate and respective MIC values were assessed using the E-test strips. The confirmatory disk diffusion methods were applied for phenotypic identification of extended-spectrum β- lactamase (ESBL production for E. coli and K. pneumoniae, according to CLSI guidelines.Results: Cultivation and re-identification of the strains yielded 858 isolates, consisting of 224 S. aureus, 148 Klebsiellaspp., 105 Serratia spp., 146 E. coli, 67 Acinetobacter spp., 38 Enterobacter spp., 95 Pseudomonas spp., 71 P.aeruginosa.35 Stenotrophomonas sp., and 8 other organisms. MRSA was detected in 37.5% of the isolates. No vancomycin-resistant or vancomycin-intermediate resistant S. aureus was detected. With the exception of Acinetobacter and Stenotrophomonas, 85% of the Gram-negative isolates were found to be susceptible in vitro to imipenem. Overall, about 61% of K. pneumoniae and 35% of E. coli isolates were ESBL producing.Conclusion: Multidrug resistant isolates

  15. Evaluation of peptide nucleic acid-mediated multiplex real-time PCR kits for rapid detection of carbapenemase genes in gram-negative clinical isolates.

    Science.gov (United States)

    Jeong, Seri; Kim, Jung Ok; Jeong, Seok Hoon; Bae, Il Kwon; Song, Wonkeun

    2015-06-01

    The emergence of clinical isolates of carbapenemase-producing microbes confers multidrug-resistance to these bacteria and renders them difficult to treat. This study was performed to evaluate peptide nucleic acid (PNA) probe-based multiplex real-time PCR kits used to detect carbapenemase genes. In total, 324 carbapenemase genes, collected from 318 gram-negative clinical isolates in 36 different hospital laboratories, were assayed to evaluate multiplex real-time PCR kits (PANAGENE; Daejeon, Korea). The nine most prevalent carbapenemase genes (KPC, OXA-48, GES, IMP, VIM, NDM, ISAba1-OXA-51, OXA-23, and OXA-58) were included in this study. The sensitivity and specificity of the multiplex real-time PCR assay to all of the carbapenemase genes were above 99.0%, except for ISAba1-OXA-51. The detection limit of the assay was 100 target copies per 25 μL of reaction volume for all of the nine genetic types of carbapenemases, and the genes were all detected in a single three-hour PCR. The assay also showed considerable efficiency (above 80.0%), stable reproducibility (coefficient of variation, below 5.0%) and a long shelf-life (more than eight months) with no cross reactivity. The developed PNA-mediated multiplex real-time PCR assay was useful for the rapid, accurate and simultaneous identification of nine carbapenemase genes in gram-negative clinical isolates, suggesting its potential to help choose the appropriate antibiotics and aid the control of carbapenemase genes.

  16. Isolation

    DEFF Research Database (Denmark)

    Agerholm, Frank Juul

    2011-01-01

    Næringsstoffet har i dette nummer sat fokus på ”velvære i vinterkulden”, ”indendørsaktiviteter” og ”fedtafgift”. I klummen vises det, at disse tre fokusområder, der for en umiddelbar betragtning måske nok synes noget uensartede, falder sammen i ét tema: Isolation!......Næringsstoffet har i dette nummer sat fokus på ”velvære i vinterkulden”, ”indendørsaktiviteter” og ”fedtafgift”. I klummen vises det, at disse tre fokusområder, der for en umiddelbar betragtning måske nok synes noget uensartede, falder sammen i ét tema: Isolation!...

  17. Isolation of cultivable microorganisms from Polish notes and coins.

    Science.gov (United States)

    Kalita, Michal; Palusińska-Szysz, Marta; Turska-Szewczuk, Anna; Wdowiak-Wróbel, Sylwia; Urbanik-Sypniewska, Teresa

    2013-01-01

    The potential role of currency in the spread of pathogenic microflora has been evaluated in many countries. In this study Polish paper notes and the coins in general circulation were assayed for the presence of cultivable bacteria and fungi. Bacterial isolates identification was based on cultural and biochemical characters and by comparison of the 16S rRNA gene sequence. Fungal isolates were recognized with biochemical and morphological criteria. Coagulase-negative staphylococci, (43.6% of the total bacterial count) including Staphylococcus saprophyticus, S. epidermidis, and S. hominis, and Enteroccus spp. (30.8% of the total bacterial count), i.e. E.faecalis, E.faecium and E. durans, were the most numerous bacterial contamination. Penicillium spp., and Aspergillus spp. were the most frequently detected moulds whereas Candida spp. was the most frequent yeast isolated from currency. A visible dependence between the banknote denomination, the physical condition of paper currency, and the number of bacteria and fungi was found. The overall count of bacteria isolated from currency was thousand-fold higher than that of fungal isolates. The total amount of bacteria and fungi recovered from the coins was approximately 2.7-fold lower than that isolated from the notes. In summary, the Polish currency notes were found to be contaminated mainly with commensal bacteria and fungi while the opportunistic pathogenic microorganisms Escherichia coli, Pseudomonas stutzeri and C. albicans were detected at a low frequency.

  18. Antimicrobial susceptibility testing of Gram-positive and -negative bacterial isolates directly from spiked blood culture media with Raman spectroscopy.

    Science.gov (United States)

    Dekter, H E; Orelio, C C; Morsink, M C; Tektas, S; Vis, B; Te Witt, R; van Leeuwen, W B

    2017-01-01

    Patients suffering from bacterial bloodstream infections have an increased risk of developing systematic inflammatory response syndrome (SIRS), which can result in rapid deterioration of the patients' health. Diagnostic methods for bacterial identification and antimicrobial susceptibility tests are time-consuming. The aim of this study was to investigate whether Raman spectroscopy would be able to rapidly provide an antimicrobial susceptibility profile from bacteria isolated directly from positive blood cultures. First, bacterial strains (n = 133) were inoculated in tryptic soy broth and incubated in the presence or absence of antibiotics for 5 h. Antimicrobial susceptibility profiles were analyzed by Raman spectroscopy. Subsequently, a selection of strains was isolated from blood cultures and analyzed similarly. VITEK®2 technology and broth dilution were used as the reference methods. Raman spectra from 67 antibiotic-susceptible strains showed discriminatory spectra in the absence or at low concentrations of antibiotics as compared to high antibiotic concentrations. For 66 antibiotic-resistant strains, no antimicrobial effect was observed on the bacterial Raman spectra. Full concordance with VITEK®2 data and broth dilution was obtained for the antibiotic-susceptible strains, 68 % and 98 %, respectively, for the resistant strains. Discriminative antimicrobial susceptibility testing (AST) profiles were obtained for all bacterial strains isolated from blood cultures, resulting in full concordance with the VITEK®2 data. It can be concluded that Raman spectroscopy is able to detect the antimicrobial susceptibility of bacterial species isolated from a positive blood culture bottle within 5 h. Although Raman spectroscopy is cheap and rapid, further optimization is required, to fulfill a great promise for future AST profiling technology development.

  19. Changing trend of antimicrobial resistance among gram-negative bacilli isolated from lower respiratory tract of ICU patients: A 5-year study

    Directory of Open Access Journals (Sweden)

    Deep Gagneja

    2011-01-01

    Full Text Available Background: Lower respiratory tract infections (LRTIs are the most frequent infections among patients in intensive care units (ICUs. Aim: To track the resistance rate among the causative agents causing LRTI in the ICU patients. Design and Settings: This is a retrospective study done in a tertiary care hospital. Materials and Methods: Transtracheal or bronchial aspirates from 2776 patients admitted to the ICU were cultured and identified, and antibiotic sensitivity was performed by standard methods. Results: Of 2776 specimens, 1233 (44.41% isolates were recovered, of which 1123 (91.07% were gram-negative bacilli (GNB and 110 (8.92% were gram-positive organisms. From 2004 to 2009, Pseudomonas aeruginosa remained the most common pathogen. In phase I, high level of resistance (79-98% was observed against all GNB. During phase II increasing trend in resistance to cephalosporins and declining trend in resistance to aminoglycosides against most GNB were observed. Multidrug resistance (resistance to three or more than three drugs was observed in 83% of total isolates. Conclusions: Gram-negative organisms are the predominant pathogens causing LRTI in ICU. The increasing trend of resistance to cephalosporins and carbapenems in gram-negative organisms is very disturbing. Judicious use of antimicrobial agents is essential to prevent the emergence of multidrug-resistant bacteria in the ICU.

  20. Mitochondrial dysfunction bridges negative affective disorders and cardiomyopathy in socially isolated rats: Pros and cons of fluoxetine.

    Science.gov (United States)

    Sonei, Nazanin; Amiri, Shayan; Jafarian, Iman; Anoush, Mahdieh; Rahimi-Balaei, Maryam; Bergen, Hugo; Haj-Mirzaian, Arya; Hosseini, Mir-Jamal

    2017-02-01

    Objectives Depression is tightly associated with cardiovascular comorbidity and accounts for high financial and social burden worldwide. Mitochondrial dysfunction contributes to the pathophysiology of depression and cardiovascular disorders; its contribution to depression-cardiovascular comorbidity has not yet been investigated. Methods Adolescent rats were subjected to 4 weeks of isolation (social isolation stress or SIS) or social conditions (control), and then they were divided into treatment (fluoxetine, 7.5 mg/kg/day for 21 days) and non-treatment groups. After different housing conditions and treatment, animals were evaluated by behavioural tests (n = 6-8) and mitochondrial assessments (n = 3) of brain and cardiac tissues. Results We found that juvenile SIS induced behavioural abnormalities and mitochondrial dysfunction in adulthood. We showed that juvenile SIS was associated with impaired respiratory chain complex, which leads to reactive oxygen species formation, oxidative damage and ATP abatement in both brain and heart. Administration of FLX (7.5 mg/kg/day) during the isolation period attenuated the effects of SIS on the brain mitochondria and behavioural abnormalities, but had little or no effect on SIS-induced mitochondrial dysfunction in cardiac tissue. Conclusions This suggests that juvenile SIS predisposes the co-occurrence of depression and cardiovascular disease through mitochondrial dysfunction and that therapeutic effect of fluoxetine is partly mediated by its effect on mitochondrial function.

  1. Nationwide German Multicenter Study on Prevalence of Antibiotic Resistance in Staphylococcal Bloodstream Isolates and Comparative In Vitro Activities of Quinupristin-Dalfopristin

    Science.gov (United States)

    von Eiff, Christof; Reinert, Ralf René; Kresken, Michael; Brauers, Johannes; Hafner, Dieter; Peters, Georg

    2000-01-01

    Antibiotic-resistant gram-positive bacteria have become an increasing problem in the last two decades. In order to evaluate the prevalence of antibiotic resistance in staphylococcal bloodstream isolates in Germany, 2,042 staphylococci collected in 21 tertiary-care hospitals were investigated during a 3-year period (March 1996 to March 1999). Altogether, 1,448 S. aureus isolates and 594 coagulase-negative staphylococci (CoNS) that comprised 13 different species were included. Furthermore, the antistaphylococcal activities of quinupristin-dalfopristin were compared with those of eight other compounds by the broth microdilution method. The rates of oxacillin resistance in Staphylococcus aureus, S. epidermidis, S. haemolyticus, and other CoNS were 13.5, 69, 90, and 34%, respectively. In oxacillin-resistant strains high rates of resistance (up to 100%) to erythromycin, clindamycin, ciprofloxacin, and gentamicin were also observed. However, no strain appeared to be resistant to vancomycin or quinupristin-dalfopristin. The streptogramin combination exhibited excellent in vitro activity against all staphylococcal species tested, regardless of the patterns of resistance to other drug classes. In terms of MICs at which 90% of the isolates are inhibited, quinupristin-dalfopristin was 2 times more active against S. aureus isolates, 4 to 16 times more active against S. haemolyticus, and 8 to 32 times more active against S. epidermidis than vancomycin or teicoplanin. PMID:10921933

  2. Diversity of culturable Gram-negative bacteria isolated from irrigation water of two rice crop regions in Southern Brazil.

    Science.gov (United States)

    Reche, Maria Helena Lima Ribeiro; Reali, Catiusca; Pittol, Michele; de Athayde Saul, Danilo; Macedo, Vera Regina Mussoi; Valiati, Victor Hugo; Machado, Vilmar; Fiuza, Lidia Mariana

    2016-06-01

    In this study, we assessed the diversity of Gram-negative bacteria found in water used for irrigation of rice crops in two growing areas of southern Brazil. Samples were collected from the main irrigation channel and field drain area. Twenty-two bacterial species were found in Cachoeirinha and 28 in Camaquã. In both areas, the most frequent bacterial families were Enterobacteriaceae and Aeromonadaceae. Differences in microbial diversity were observed in both study areas. Thirty-five Gram-negative species were identified; however, only 15 were common in both locations. In addition, there were found pathogenic and drug-resistant species, such as Acinetobacter sp., Brucella spp., and Chryseobacterium meningosepticum. This study demonstrates the existence of a number of pathogenic species in aquatic ecosystems analyzed in three consecutive crop years, especially water used for rice production.

  3. Antimicrobial resistance and detection of mecA and blaZ genes in coagulase-negative Staphylococcus isolated from bovine mastitis

    Directory of Open Access Journals (Sweden)

    Lidiane C. Soares

    2012-08-01

    Full Text Available The present study evaluated the pheno- and genotypical antimicrobial resistance profile of coagulase-negative Staphylococcus (CNS species isolated from dairy cows milk, specially concerning to oxacillin. Of 100 CNS isolates, the S. xylosus was the prevalent species, followed by S. cohnii, S. hominis, S. capitis and S. haemolyticus. Only 6% were phenotypically susceptible to the antimicrobial agents tested in disk diffusion assay. Penicillin and ampicillin resistance rates were significantly higher than others antimicrobials. Four isolates were positive to mecA gene (4%, all represented by the S. xylosus species. The blaZ gene was detected in 16% of the isolates (16/100. It was noticed that all mecA + were also positive to this gene and the presence of both genes was correlated to phenotypic beta-lactamic resistance. We conclude that CNS species from bovine milk presented significantly distinct antimicrobial resistance profiles, evaluated by phenotypic and genotypic tests, which has implications for treatment and management decisions.

  4. Isolation of Mycoplasma gallopavonis from free-ranging wild turkeys in coastal North Carolina seropositive and culture-negative for Mycoplasma gallisepticum.

    Science.gov (United States)

    Cobb, D T; Ley, D H; Doerr, P D

    1992-01-01

    Serum samples and choanal cleft swabs were collected from livetrapped and hunter killed wild turkeys (Meleagris gallopavo) from Martin and Bertie counties, North Carolina (USA). Sera were tested for antibodies to Mycoplasma gallisepticum, Mycoplasma synoviae and Mycoplasma meleagridis by hemagglutination inhibition (HI). Sera from 33% (five of 15) of livetrapped turkeys were positive for antibodies to M. gallisepticum by HI, and all were negative for antibodies to M. synoviae and M. meleagridis. Choanal cleft swabs from 22 livertrapped and five hunter killed wild turkeys cultured in Frey's broth medium resulted in 23 mycoplasma isolations. Using direct immunofluorescence, 74% (17/23) were M. gallopavonis, and 26% (six of 23) were unidentified; no isolate was identified as M. gallisepticum, M. synoviae or M. meleagridis.

  5. A subpopulation of large granular von Willebrand Ag negative and CD105 positive endothelial cells, isolated from abdominal aortic aneurysms, overexpress ICAM-1 and Fas antigen.

    Science.gov (United States)

    Páez, Araceli; Archundia, Abel; Méndez Cruz, René; Rodríguez, Emma; López Marure, Rebeca; Masso, Felipe; Aceves, José Luis; Flores, Leopoldo; Montaño, Luis F

    2002-01-01

    The aim of this work was to determine whether there is a pre-established basal condition of the endothelial cells isolated from aortic abdominal aneurysm that might augment immune effector mechanisms and thus provide us an insight into the possible causes of aneurysm rupture. Endothelial cells isolated from saccular aortic aneurysm fragments were analyzed by cytofluorometry for the expression of different immune response-related molecules. Our results showed that there is a subpopulation of granule-rich, CD105 positive and von Willebrand antigen negative endothelial cells that have an enhanced basal expression of ICAM-1, and Fas antigen, but, interestingly, no apoptotic bodies were detected. Control endothelial cells derived from healthy areas of the same abdominal aortas did not show such enhanced expression. We conclude that in the endothelium that lines abdominal aorta aneurysms there is, at least, one endothelial cell subpopulation with an apparent inhibition of programmed cell death and in a proinflammatory activation status.

  6. Enacted support's links to negative affect and perceived support are more consistent with theory when social influences are isolated from trait influences.

    Science.gov (United States)

    Lakey, Brian; Orehek, Edward; Hain, Kate L; Van Vleet, Meredith

    2010-01-01

    Social support theory typically explains perceived support's link to mental health as reflecting the role of specific supportive actions (i.e., enacted support).Yet enacted support typically is not linked to mental health and perceived support as predicted by theory. The links are examined among enacted support, affect, and perceived support when links reflected (a) aspects of support and affect that generalized across relationship partners and time (i.e., trait influences) and (b) aspects that reflected specific relationship partners (i.e., social influences). Multivariate generalizability analyses indicated that enacted support was linked to low negative affect as predicted by theory only when correlations reflected social influences. When correlations reflected trait influences, enacted support was linked to high negative affect. Furthermore, perceived and enacted support were strongly linked when correlations reflected social influences but not trait influences. Thus, findings for enacted support fit social support theory better when social influences were isolated from trait influences.

  7. Array based detection of antibiotic resistance genes in Gram negative bacteria isolated from retail poultry meat in the UK and Ireland.

    Science.gov (United States)

    McNeece, Grainne; Naughton, Violetta; Woodward, Martin J; Dooley, James S G; Naughton, Patrick J

    2014-06-02

    The use of antibiotics in birds and animals intended for human consumption within the European Union (EU) and elsewhere has been subject to regulation prohibiting the use of antimicrobials as growth promoters and the use of last resort antibiotics in an attempt to reduce the spread of multi-resistant Gram negative bacteria. Given the inexorable spread of antibiotic resistance there is an increasing need for improved monitoring of our food. Using selective media, Gram negative bacteria were isolated from retail chicken of UK-Intensively reared (n=27), Irish-Intensively reared (n=19) and UK-Free range (n=30) origin and subjected to an oligonucleotide based array system for the detection of 47 clinically relevant antibiotic resistance genes (ARGs) and two integrase genes. High incidences of β-lactamase genes were noted in all sample types, acc (67%), cmy (80%), fox (55%) and tem (40%) while chloramphenicol resistant determinants were detected in bacteria from the UK poultry portions and were absent in bacteria from the Irish samples. Denaturing Gradient Gel Electrophoresis (DGGE) was used to qualitatively analyse the Gram negative population in the samples and showed the expected diversity based on band stabbing and DNA sequencing. The array system proved to be a quick method for the detection of antibiotic resistance gene (ARG) burden within a mixed Gram negative bacterial population.

  8. enterotoxigenicity and drug sensitivity of staphylococci from children ...

    African Journals Online (AJOL)

    hi-tech

    2003-12-12

    Dec 12, 2003 ... children aged five years and below suffering from sporadic diarrhoea and their antibiotic susceptibility ... were isolated exclusive of other pathogens were studied further. Strains ... isolate under test and it was indicated by no change in colour .... Keersmaecker, P.H. Staphylococcus hyicus in skin lesions.

  9. Potential sources of bacteria that are isolated from contact lenses during wear.

    Science.gov (United States)

    Willcox, M D; Power, K N; Stapleton, F; Leitch, C; Harmis, N; Sweeney, D F

    1997-12-01

    The aim of this paper was to determine the possible contamination sources of contact lenses during wear. Potential sources of the microbiota that colonized hydrogel contact lenses during wear were examined. The microorganisms that colonize contact lenses were grown, identified, and compared to those microorganisms that colonized the lower lid margins, upper bulbar conjunctiva, hands, and contact lens cases of contact lens wearers. In addition, the incidence of contamination of the domestic water supply in the Sydney area was obtained, and this was compared to the incidence of colonization of contact lenses by microorganisms in general and gram-negative bacteria in particular. There was a wide diversity of bacteria that were isolated from each site sampled. Coagulase-negative staphylococci and Propionibacterium spp. were the most common isolates from all ocular sites examined, and constituted the normal ocular microbiota. Other bacteria, including members of the families Enterobacteriaceae and Pseudomonadaceae, were isolated infrequently from all sites, but most frequently from contact lens cases. Statistical analysis revealed that there was a correlation between the isolation of bacteria from the contact lens and the lower lid margin (p microbiota. A correlation was also noted between the colonization of contact lenses by gram-negative bacteria and contamination of the domestic water supply. This study has demonstrated that the likely route for the normal ocular microbiota colonizing contact lenses is via the lid margins, whereas colonization by gram-negative bacteria, including potential agents of microbial keratitis, is likely to be from the domestic water supply.

  10. Prevalence of Carbapenem Resistant Non-Fermenting Gram Negative Bacterial Infection and Identification of Carbapenemase Producing NFGNB Isolates by Simple Phenotypic Tests.

    Science.gov (United States)

    Esther, Jane; Edwin, Diego; Uma

    2017-03-01

    Non-Fermenting Gram negative bacilli (NFGNB) are emerging multi-drug resistant pathogens causing nosocomial infections. In recent years, carbapenem resistance in NFGNB has increased due to a variety of drug resistance mechanisms, the most common being production of carbapenemases. To detect carbapenemase and metallo-β-lactamase (MBL) production in NFGNBs by four phenotypic tests and to compare the various phenotypic methods for detection of carbapenemase and MBL production in nosocomial NFGNB isolates. It is a cross sectional study carried out in the department of Microbiology, Chennai Medical College Hospital and Research Center, Irungalur, Trichy between January 2015 and December 2015. Out of the 598 NFGNB isolated from all the 5402 heterogenous clinical samples that were processed, 52 (8.7%) NFGNB showed resistance or intermediate sensitivity to meropenem as tested by disc diffusion assay. All the 52 isolates were subjected to four different phenotypic tests for carbapenemase and MBL detection, which included Modified Hodge Test (MHT), Meropenem-EDTA Disc Synergy (EDS) test, Meropenem-EDTA Combined Disc Test (CDT) and Growth on CHROMagar KPC. Among the 52 isolates, 29 (55.77%) were MHT positive and 49 (94.23%) were positive for growth on CHROMEagar KPC which were identified as carbapenemase producers. 44 (84.61%) were EDS positive and 46 (88.46%) were CDT positive which were identified as metallo-β-lactamase producers. The presence of these resistant bugs strongly suggests the need to prevent their further spread by implementation of strict infection control measures and regular surveillance to check their outcome. Growth on CHROMagar KPC is the test that has picked up more number of carbapenem resistant isolates as carbapenemase and metallo-β-lactamase producers among the four tests. It is also cheap and easy to perform, making it the most reliable test for routine screening of carbapenemase and MBL producers in clinical laboratories.

  11. Kanagawa-negative, tdh- and trh-positive Vibrio parahaemolyticus isolated from fresh oysters marketed in Fortaleza, Brazil.

    Science.gov (United States)

    Vieira, Regine H S F; Costa, Renata A; Menezes, Francisca G R; Silva, Giselle C; Theophilo, Grace N D; Rodrigues, Dália P; Maggioni, Rodrigo

    2011-08-01

    Between October 2008 and June 2009, 15 samples of 10 live oysters each (Crassostrea rhizophorae) measuring 8.31-10.71 cm were purchased from a restaurant on the seashore of Fortaleza, Brazil. The Vibrio count ranged from 75 (estimated) to 43,500 CFU/g. Fourteen species were identified among the 56 isolated Vibrio strains, with V. parahaemolyticus as the most prevalent. Two of the 17 V. parahaemolyticus strains were urease-positive and tdh- and trh-positive on multiplex PCR, but neither produced β-hemolysis halos in Wagatsuma agar. Thus, fresh oysters served in natura in Fortaleza, Brazil, were found to contain Vibrio strains known to cause gastroenteritis in humans.

  12. Study of frequency of bacteria isolated from blood culture and their antibiotic susceptibility pattern in a university hospital in Tehran

    Directory of Open Access Journals (Sweden)

    Hoorieh Saderi

    2009-12-01

    Full Text Available Introduction: Determining frequency of bacteria, isolated from blood culture and their antibiotic susceptibility patterns, has epidemiological significance and can help in selecting empirical therapy. This study was aimed to assess, the frequency of bacteria isolated from blood culture of patients suspected to bacteremia and their antibiotic susceptibility patterns. Methods: Culture of blood and determination of antibiotic susceptibility was done by standard methods. In this study, a variety of isolated bacteria types, antibiotic susceptibility, as well as age, sex and type of admission of patients were analyzed in a university hospital from 21 March, 2006 to 20 March, 2007. Results: During one year, blood culture was done for 5116 patients and bacteria were isolated in 912 cases (17.8%. Three most frequently groups of bacteria in blood cultures of patients were non-fermentative gram negative bacteria (Pseudomonas and Acintobacter spp, coliforms (Escherichia coli and enterobacter and klebsiella spp. and coagulase negative staphylococci, respectively, which were isolated in 63.4%, 17.0% and 12.8% of patients, and constituted 93.2% of positive blood cultures. Higher resistance was shown in bacteria isolated from inpatients compare to outpatients. Conclusion: This study showed the influence of age, sex and type of admission (outpatient or inpatient in a variety of isolated bacteria in blood culture. The result of this study were the same as the other studies in Iran and other countries in respect of the variety of isolated bacteria and antibiotic susceptibility and show increase of antibiotic resistance in these bacteria.

  13. Presence of superantigen genes and antimicrobial resistance in Staphylococcus isolates obtained from the uteri of dairy cows with clinical endometritis.

    Science.gov (United States)

    Zhao, J-L; Ding, Y-X; Zhao, H-X; He, X-L; Li, P-F; Li, Z-F; Guan, H; Guo, X

    2014-10-11

    Clinical endometritis is an important disease of dairy cattle and results in decreased reproductive performance. This disease is caused by contamination of the uterus with a broad spectrum of microorganisms after calving. In this study, staphylococcal isolates from the uterus of dairy cows with clinical endometritis were tested for their distribution of superantigen (SAg) genes and antimicrobial resistance. Between the 127 staphylococcal isolates collected in this study, 10 species were identified. The predominant strain identified was Staphylococcus aureus (n=53), followed by Staphylococcus saprophyticus (n=38) and Staphylococcus chromogenes (n=22). PCR analysis demonstrated that most isolates (63.0 per cent) harboured at least one SAg gene. The most commonly observed SAg gene and genotype was selj (38.6 per cent) and sec-selj-seln (24.0 per cent), respectively. Most isolates were resistant to penicillin (79.5 per cent), ampicillin (71.7 per cent), erythromycin (56.7 per cent), and tetracycline (52.0 per cent). PCR analysis demonstrated that the antimicrobial resistance determinants ermA, ermB, ermC, tetK, tetM and blaZ were detected in 0 per cent, 44.4 per cent, 51.4 per cent, 68.2 per cent, 13.6 per cent and 86.1 per cent of the erythromycin, tetracycline and β-lactam resistant isolates, respectively. There were 22 (17.3 per cent of all isolates) coagulase-negative staphylococci shown to be methicillin resistant. In the methicillin-resistant isolates, significant resistances to ampicillin, erythromycin and penicillin were observed (P<0.01). The results of this study demonstrate that staphylococci recovered from dairy cows with clinical endometritis contain an extensive and complex prevalence of SAg genes. Significant resistances to antibiotics were also seen, highlighting the need for the rational appliance of antibiotics in veterinary medicine.

  14. Antibiotic sensitivity pattern of gram negative bacilli isolated from the lower respiratory tract of ventilated patients in the intensive care unit

    Directory of Open Access Journals (Sweden)

    Goel Nidhi

    2009-01-01

    Full Text Available Background: Lower respiratory tract infections (LRTIs are the most frequent infections among patients in Intensive care units (ICUs. Aims: To know the bacterial profile and determine the antibiotic susceptibility pattern of the lower respiratory tract isolates from patients admitted to the ICU. Settings and Design: Tertiary care hospital, retrospective study. Materials and Methods: Transtracheal or bronchial aspirates from 207 patients admitted to the ICU were cultured, identified, and antibiotic sensitivity was performed by standard methods. Statistical Analysis Used: SPSS software was used for calculation of % R of 95% confidence interval (CI. Results: Of 207 specimens, 144 (69.5% were culture positive and 63 (30.4% showed no growth. From 144 culture positives, 161 isolates were recovered, of which 154 (95.6% were Gram negative bacilli (GNB. In 17 (11.0% patients, two isolates per specimen were recovered. The most common GNB in order of frequency were Pseudomonas aeruginosa (35%, Acinetobacter baumannii (23.6%, and Klebsiella pneumoniae (13.6%. A very high rate of resistance (80-100% was observed among predominant GNB to ciprofloxacin, ceftazidime, co-trimoxazole, and amoxycillin/clavulanic acid combination. Least resistance was noted to meropenem and doxycycline. Conclusion: Nonfermenters are the most common etiological agents of LRTIs in ICU. There is an alarmingly high rate of resistance to cephalosporin and β-lactam-β-lactamase inhibitor group of drugs. Meropenem was found to be the most sensitive drug against all GNB. Acinetobacter and Klebsiella spp. showed good sensitivity to doxycycline.

  15. Conditional probability analysis of multidrug resistance in Gram-negative bacilli isolated from tertiary medical institutions in South Korea during 1999-2009.

    Science.gov (United States)

    Kim, Yong-Hak

    2016-01-01

    Multidrug resistance of Gram-negative bacilli is a major problem globally. However, little is known about the combined probability of resistance to various antibiotics. In this study, minimum inhibitory concentrations of widely used antibiotics were determined using clinical isolates of Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Acinetobacter baumannii, randomly chosen from strain collections created during 1999-2009 in tertiary medical institutions in Seoul, South Korea. To analyze combined efficacy of antibiotics against a subgroup of isolates, conditional probabilities were determined based on arbitrary, non-independent patterns of antimicrobial susceptibility and resistance. Multidrug resistance, defined as resistance to three or more classes of antibiotics, was observed in the following order: A. baumannii (96%), P. aeruginosa (65%), E. coli (52%), and K. pneumoniae (7%). A. baumannii strains resistant to gentamicin were found to be resistant to a number of antibiotics, except for colistin and polymyxin B. Resistance to gentamicin following exposure to this antibiotic was highly likely to lead to multidrug resistance in all four microbes. This study shows a causal relationship between gentamicin resistance and the prevalence of multidrug resistance in clinical isolates of Gramnegative bacilli in South Korea during 1999-2009 and suggests the importance of prudent use of gentamicin in hospitals.

  16. Fibronectin binding to protein A-containing staphylococci.

    OpenAIRE

    Doran, J E; Raynor, R H

    1981-01-01

    Fibronectin (Fn) was found to bind to protein A-containing isolates of Staphylococcus aureus, but not to mutant strains devoid of this protein nor to clinical isolates of S. epidermidis. Fn was purified from human plasma by affinity chromatography on gelatin-Sepharose. After elution with 4 M urea, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified material detected no immunoglobulin contamination. This purified Fn was radiolabeled with 125I and used in binding assays. Quant...

  17. Isolation of Trichoderma reesei pyrG Negative Mutant by UV Mutagenesis and Its Application in Transformation

    Institute of Scientific and Technical Information of China (English)

    LONG Hao; WANG Tian-hong; ZHANG Ying-kuan

    2008-01-01

    Two uridine auxotrophic mutants of Trichoderma reesei were isolated by resistance to 5-fluoroorotic acid after UV mutagenesis.One mutant,called M23,was complemented with the Aspergillus niger pyrG gene carded by plasmid pAB4-1.A mutated pyrG gene of M23 was cloned and DNA sequencing analysis indicated that a cytosine was inserted into the 934-939 oligo dC position of the pyrG coding region,resulted in a frameshift mutation.Transformation efficiency was approximately 200-300 transformants per microgram of DNA with plasmid pAB4-1.Stable transformants were obtained by monosporic culture and showed to be prototroph after successive propagation.Vitreoscilla hemoglobin expression plasmid pUCVHb was cotransformed with plasmid pAB4-1 and attained a transformation efficiency of 71.8% or of 26.1% with pAN7-1.Southern blot analysis of the transformants demonstrated that plasmid pUCVHb was integrated into the chromosomal DNA.The experimental results demonstrated that the pyrG-based system was more efficient and timesaving than the conventional hygromycin B resistance-based transformation system.

  18. Effect of BiTek agar on lysostaphin susceptibility of staphylococci.

    OpenAIRE

    Langlois, B E; Dawson, K.; Akers, K

    1990-01-01

    Staphylococci which were considered to be lysostaphin susceptible on P agar containing Bacto-Agar showed different degrees of resistance to lysostaphin when tested on P agar made with BiTek agar. As a result, lysostaphin-susceptible strains were misidentified as lysostaphin-resistant strains.

  19. Identification of methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from burn patients by multiplex PCR.

    Science.gov (United States)

    Montazeri, Effat Abbasi; Khosravi, Azar Dokht; Jolodar, Abbas; Ghaderpanah, Mozhgan; Azarpira, Samireh

    2015-05-01

    Methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MRCoNS) as important human pathogens are causes of nosocomial infections worldwide. Burn patients are at a higher risk of local and systemic infections with these microorganisms. A screening method for MRSA by using a multiplex polymerase chain reaction (PCR) targeting the 16S ribosomal RNA (rRNA), mecA, and nuc genes was developed. The aim of the present study was to investigate the potential of this PCR assay for the detection of MRSA strains in samples from burn patients. During an 11-month period, 230 isolates (53.11%) of Staphylococcus spp. were collected from burn patients. The isolates were identified as S. aureus by using standard culture and biochemical tests. DNA was extracted from bacterial colonies and multiplex PCR was used to detect MRSA and MRCoNS strains. Of the staphylococci isolates, 149 (64.9%) were identified as S. aureus and 81 (35.21%) were described as CoNS. Among the latter, 51 (62.97%) were reported to be MRCoNS. From the total S. aureus isolates, 132 (88.6%) were detected as MRSA and 17 (11.4%) were methicillin-susceptible S. aureus (MSSA). The presence of the mecA gene in all isolates was confirmed by using multiplex PCR as a gold standard method. This study presented a high MRSA rate in the region under investigation. The 16S rRNA-mecA-nuc multiplex PCR is a good tool for the rapid characterization of MRSA strains. This paper emphasizes the need for preventive measures and choosing effective antimicrobials against MRSA and MRCoNS infections in the burn units. Copyright © 2014 Elsevier Ltd and ISBI. All rights reserved.

  20. Antibacterial activities of almond skins on cagA-positive and-negative clinical isolates of Helicobacter pylori

    Science.gov (United States)

    2013-01-01

    Background Helicobacter pylori is known to be a gastric pathogen of humans. Eradication regimens for H. pylori infection have some side effects, compliance problems, relapses, and antibiotic resistance. Therefore, the need for alternative therapies for H. pylori infections is of special interest. We have previously shown that polyphenols from almond skins are active against a range of food-borne pathogens. The aim of this study was to evaluate the antibacterial effects of natural almond skins before and after simulated human digestion and the pure flavonoid compounds epicatechin, naringenin and protocatechuic acid against H. pylori. Results H. pylori strains were isolated from gastric biopsy samples following standard microbiology procedures. Also, cagA and vacA genes were identified using PCR. Susceptibility studies on 34 strains of H. pylori, including two reference strains (ATCC 43504, ATCC 49503), were performed by the standard agar dilution method. Natural almond skin was the most effective compound against H. pylori (MIC range, 64 to 128 μg/ml), followed by natural skin post gastric digestion (MIC range, 128 to 512 μg/ml), and natural almond skin post gastric plus duodenal digestion (MIC range, 256 to 512 μg/ml). Amongst the pure flavonoid compounds, protocatechuic acid showed the greatest activity (MIC range, 128 to 512 μg/ml) against H. pylori strains. Conclusions Polyphenols from almond skins were effective in vitro against H. pylori, irrespective of genotype status and could therefore be used in combination with antibiotics as a novel strategy for antibiotic resistance. PMID:23659287

  1. Isolation and Characterization of Escherichia coli Sequence Type 131 and Other Antimicrobial-Resistant Gram-Negative Bacilli from Clinical Stool Samples from Veterans.

    Science.gov (United States)

    Mohamed, Muhanad; Clabots, Connie; Porter, Stephen B; Thuras, Paul; Johnson, James R

    2016-08-01

    Emerging multidrug-resistant (MDR) Gram-negative bacilli (GNB), including Escherichia coli sequence type 131 (ST131) and its resistance-associated H30 subclone, constitute an ever-growing public health threat. Their reservoirs and transmission pathways are incompletely defined. To assess diarrheal stools as a potential reservoir for ST131-H30 and other MDR GNB, we cultured 100 clinical stool samples from a Veterans Affairs Medical Center clinical laboratory (October to December 2011) for fluoroquinolone- and extended-spectrum cephalosporin (ESC)-resistant E. coli and other GNB, plus total E. coli We then characterized selected resistant and susceptible E. coli isolates by clonal group, phylogenetic group, virulence genotype, and pulsotype and screened all isolates for antimicrobial resistance. Overall, 79 of 100 stool samples yielded GNB (52 E. coli; 48 other GNB). Fifteen samples yielded fluoroquinolone-resistant E. coli (10 were ST131, of which 9 were H30), 6 yielded ESC-resistant E. coli (2 were ST131, both non-H30), and 31 yielded susceptible E. coli (1 was ST131, non-H30), for 13 total ST131-positive samples. Fourteen non-E. coli GNB were ESC resistant, and three were fluoroquinolone resistant. Regardless of species, almost half (46%) of the fluoroquinolone-resistant and/or ESC-resistant non-E. coli GNB were resistant to at least three drug classes. Fecal ST131 isolates closely resembled reference clinical ST131 isolates according to virulence genotypes and pulsed-field gel electrophoresis (PFGE) profiles. Thus, a substantial minority (30%) of veterans with diarrhea who undergo stool testing excrete antibiotic-resistant GNB, including E. coli ST131. Consequently, diarrhea may pose transmission risks for more than just diarrheal pathogens and may help disseminate clinically relevant ST131 strains and other MDR GNB within hospitals and the community. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  2. Impact of Ciprofloxacin and Clindamycin Administration on Gram-Negative Bacteria Isolated from Healthy Volunteers and Characterization of the Resistance Genes They Harbor.

    Science.gov (United States)

    Card, Roderick M; Mafura, Muriel; Hunt, Theresa; Kirchner, Miranda; Weile, Jan; Rashid, Mamun-Ur; Weintraub, Andrej; Nord, Carl Erik; Anjum, Muna F

    2015-08-01

    The aim of this study was to assess the impact of ciprofloxacin, clindamycin, and placebo administration on culturable Gram-negative isolates and the antibiotic resistance genes they harbor. Saliva and fecal samples were collected from healthy human volunteers before and at intervals, up to 1 year after antibiotic administration. Samples were plated on selective and nonselective media to monitor changes in different colony types or bacterial species. Following ciprofloxacin administration, there was a decrease of Escherichia coli in feces and after clindamycin administration a decrease of Bacteroides in feces and Leptotrichia in saliva, which all returned to pretreatment levels within 1 to 4 months. Ciprofloxacin administration also resulted in an increase in ciprofloxacin-resistant Veillonella in saliva, which persisted for 12 months. Additionally, 949 aerobic and anaerobic isolates purified from ciprofloxacin- and clindamycin-containing plates were screened for the presence of resistance genes. Resistance gene carriage was widespread in isolates from all three treatment groups, and no association was observed between genes and antibiotic administration. Although the anaerobic component of the microbiota was not a major reservoir of aerobe-associated antimicrobial resistance (AMR) genes, we detected the sulfonamide resistance gene sul2 in anaerobic isolates. The longitudinal nature of the study allowed identification of distinct Escherichia coli clones harboring multiple resistance genes, including one carrying an extended-spectrum β-lactamase blaCTX-M group 9 gene, which persisted in the gut for up to 4 months. This study provided insight into the effects of antibiotic administration on healthy microbiota and the diversity of resistance genes harbored therein.

  3. Comparative analysis of antibiotic resistance characteristics of Gram-negative bacteria isolated from laying hens and eggs in conventional and organic keeping systems in Bavaria, Germany.

    Science.gov (United States)

    Schwaiger, K; Schmied, E-M V; Bauer, J

    2008-09-01

    By investigating the prevalence and resistance characteristics of Gram-negative bacteria from organic and conventional kept laying hens against 31 (Campylobacter: 29) different antibiotics using the microdilution method, we determined to what extent different keeping systems influence bacterial resistance patterns. For this purpose, samples from 10 organic and 10 conventional flocks in Bavaria (Germany) were investigated four times between January 2004 and April 2005. Altogether, 799 cloacal swabs and 800 eggs (contents and shells) were examined. The bacterial investigation performed with standardized cultural methods showed prevalence for all bacteria groups in about the same order of magnitude in the two different keeping systems: Salmonella spp. 3.5% (organic ([org])) versus 1.8% (conventional ([con])); Campylobacter spp. 34.8%(org) versus 29.0%(con) and E. coli 64.4%(org) versus 69.0%(con). Coliforms (Citrobacter, Enterobacter, Pantoea) were only isolated in single cases. In eggs, generally less bacteria were detected, predominantly Escherichia; Salmonella and Campylobacter were only scarcely isolated. Salmonella enterica ssp. enterica serovar Typhimurium (n=10) were resistant to up to nine, S. of the serogroup B (n=4) up to six antibiotics. All tested Salmonella (n=23) proved to be resistant to spectinomycin. Escherichia coli (n=257(org) and 276(con)) from organic layers showed significant lower resistance rates and higher rates of susceptible isolates to nine agents, namely amoxicillin/clavulanic acid, ampicillin, cefaclor, cefoxitin, cefuroxime, doxycycline, mezlocillin, neomycin and piperacillin. In contrast, only two antibiotics turned out to be more effective in conventional isolates (gentamicin and tobramycin). In the case of Campylobacter jejuni (n=118(org) and 99(con)), statistically significantly better rates were observed for isolates from organic flocks concerning imipenem and amoxicillin/clavulanic acid, whereas fosfomycin was more potent in

  4. Antimicrobial susceptibilities of aerobic and facultative gram-negative bacilli isolated from Chinese patients with urinary tract infections between 2010 and 2014.

    Science.gov (United States)

    Yang, Qiwen; Zhang, Hui; Wang, Yao; Xu, Zhipeng; Zhang, Ge; Chen, Xinxin; Xu, Yingchun; Cao, Bin; Kong, Haishen; Ni, Yuxing; Yu, Yunsong; Sun, Ziyong; Hu, Bijie; Huang, Wenxiang; Wang, Yong; Wu, Anhua; Feng, Xianju; Liao, Kang; Luo, Yanping; Hu, Zhidong; Chu, Yunzhuo; Lu, Juan; Su, Jianrong; Gui, Bingdong; Duan, Qiong; Zhang, Shufang; Shao, Haifeng; Badal, Robert E

    2017-03-06

    The objective of this study was to investigate the distribution and susceptibility of aerobic and facultative Gram-negative bacilli isolated from Chinese patients with UTIs collected within 48 h (community acquired, CA) or after 48 h (hospital acquired, HA) of hospital admission. From 2010 to 2014, the minimum inhibitory concentrations (MICs) of 12 antibiotics for 4,332 aerobic and facultative Gram-negative bacilli, sampled in 21 hospitals in 16 cities, were determined by the broth microdilution method. Enterobacteriaceae composed 88.5% of the total isolates, with Escherichia coli (E. coli) (63.2%) the most commonly isolated species, followed by Klebsiella pneumoniae (K. pneumoniae) (12.2%). Non-Enterobacteriaceae accounted for only 11.5% of all isolates and included mainly Pseudomonas aeruginosa (P. aeruginosa) (6.9%) and Acinetobacter baumannii (A. baumannii) (3.3%). Among the antimicrobial agents tested, the susceptibility rates of E.coli to the two carbapenems, ertapenem and imipenem as well as amikacin and piperacillin-tazobactam ranged from 92.5 to 98.7%. Against K. pneumonia, the most potent antibiotics were imipenem (92.6% susceptibility), amikacin (89.2% susceptibility) and ertapenem (87.9% susceptibility). Although non-Enterobacteriaceae did not show high susceptibilities to the 12 common antibiotics, amikacin exhibited the highest in vitro activity against P. aeruginosa over the 5-year study period, followed by piperacillin-tazobactam, imipenem, ceftazidime, cefepime, ciprofloxacin, and levofloxacin. The Extended Spectrum Beta-Lactamase (ESBL) rates decreased slowly during the 5 years in E. coli from 68.6% in 2010 to 59.1% in 2014, in K. pneumoniae from 59.7 to 49.2%, and in Proteus mirabilis (P. mirabilis) from 40.0 to 26.1%. However, the ESBL rates were different in 5 regions of China (Northeast, North, East, South and Middle-China). E. coli and K. pneumonia were the major pathogens causing UTIs and carbapenems and amikacin retained the highest

  5. Fourier Transform Infrared Spectroscopy for Rapid Identification of Nonfermenting Gram-Negative Bacteria Isolated from Sputum Samples from Cystic Fibrosis Patients▿

    Science.gov (United States)

    Bosch, Alejandra; Miñán, Alejandro; Vescina, Cecilia; Degrossi, José; Gatti, Blanca; Montanaro, Patricia; Messina, Matías; Franco, Mirta; Vay, Carlos; Schmitt, Juergen; Naumann, Dieter; Yantorno, Osvaldo

    2008-01-01

    The accurate and rapid identification of bacteria isolated from the respiratory tract of patients with cystic fibrosis (CF) is critical in epidemiological studies, during intrahospital outbreaks, for patient treatment, and for determination of therapeutic options. While the most common organisms isolated from sputum samples are Pseudomonas aeruginosa, Staphylococcus aureus, and Haemophilus influenzae, in recent decades an increasing fraction of CF patients has been colonized by other nonfermenting (NF) gram-negative rods, such as Burkholderia cepacia complex (BCC) bacteria, Stenotrophomonas maltophilia, Ralstonia pickettii, Acinetobacter spp., and Achromobacter spp. In the present study, we developed a novel strategy for the rapid identification of NF rods based on Fourier transform infrared spectroscopy (FTIR) in combination with artificial neural networks (ANNs). A total of 15 reference strains and 169 clinical isolates of NF gram-negative bacteria recovered from sputum samples from 150 CF patients were used in this study. The clinical isolates were identified according to the guidelines for clinical microbiology practices for respiratory tract specimens from CF patients; and particularly, BCC bacteria were further identified by recA-based PCR followed by restriction fragment length polymorphism analysis with HaeIII, and their identities were confirmed by recA species-specific PCR. In addition, some strains belonging to genera different from BCC were identified by 16S rRNA gene sequencing. A standardized experimental protocol was established, and an FTIR spectral database containing more than 2,000 infrared spectra was created. The ANN identification system consisted of two hierarchical levels. The top-level network allowed the identification of P. aeruginosa, S. maltophilia, Achromobacter xylosoxidans, Acinetobacter spp., R. pickettii, and BCC bacteria with an identification success rate of 98.1%. The second-level network was developed to differentiate the four

  6. Activation of Natural Killer Cells in Patients with Chronic Bone and Joint Infection due to Staphylococci Expressing or Not the Small Colony Variant Phenotype

    Directory of Open Access Journals (Sweden)

    Sébastien Viel

    2014-01-01

    Full Text Available Chronic bone and joint infections (BJI are devastating diseases. Relapses are frequently observed, as some pathogens, especially staphylococci, can persist intracellularly by expressing a particular phenotype called small colony variant (SCV. As natural killer (NK cells are lymphocytes specialized in the killing of host cells infected by intracellular pathogens, we studied NK cells of patients with chronic BJI due to staphylococci expressing or not SCVs (10 patients in both groups. Controls were patients infected with other bacteria without detectable expression of SCVs, and healthy volunteers. NK cell phenotype was evaluated from PBMCs by flow cytometry. Degranulation capacity was evaluated after stimulation with K562 cells in vitro. We found that NK cells were activated in terms of CD69 expression, loss of CD16 and perforin, in all infected patients in comparison with healthy volunteers, independently of the SCV phenotype. Peripheral NK cells in patients with chronic BJI display signs of recent activation and degranulation in vivo in response to CD16-mediated signals, regardless of the type of bacteria involved. This could involve a universal capacity of isolates responsible for chronic BJI to produce undetectable SCVs in vivo, which might be a target of future intervention.

  7. ORIGINAL ARTICLE: Detection of β-Lactamase Activity in Various Clinical Bacterial Isolates by Three Different Methods and its Correlation with Drug Resistance.

    Directory of Open Access Journals (Sweden)

    Sanjay M Wavare

    2012-07-01

    Full Text Available Background: β-lactams such as penicillins are the most widely used antibiotics, and β-lactamases are the greatest source of resistance to penicillins. Aims and Objectives: To study β-lactamase production in clinical isolates of family Enterobacteriaceae, P. aeruginosa and Staphylococci by three different methods and to correlate its potential with drug resistance; with an endeavour to evaluate convenient and economical method duly supported by relevant Minimum Inhibitory Concentration (MIC studies. Material and Methods: Total 240 clinical isolates (Gram-negative bacilli-191, staphylococci-49 were subjected to antimicrobial susceptibility testing by Kirby-Bauer disk diffusion method and MIC for ampicillin and penicillin was determined by agar dilution method. β-lactamase was detected by broth acidometric, iodometric cell suspension and microbiological method. Results: Multidrug resistance was observed in more than 90% isolates. One hundred and ninety Gram-negative bacilli were resistant to ampicillin and 47 staphylococcal isolates were resistant to both penicillin and ampicillin. Though microbiological method gave highest positive results 210 (87.5%, iodometric method could detect β-lactamase in apparently sensitive isolates as well giving satisfactory [207 (86.25%] comparable results. Conclusion: In view of the noted bacterial resistance, tests for β-lactamase should be carried out on a routine basis for an early implementation of appropriate antimicrobial therapy. Iodometric method is eminently convenient, economical and reliable method. Isolates showing MIC <0.125µg/ml for penicillin and MIC <8µg/ml for ampicillin should be checked for β-lactamase production.

  8. Prevalence and antibiotic sensitivity of Danish versus other European bacterial isolates from intensive care and hematology/oncology units

    DEFF Research Database (Denmark)

    Fomsgaard, A; Høiby, Niels; Friis, H M

    1995-01-01

    . Bacterial isolates from 794 Danish patients were tested and compared to 8,625 isolates from European patients. The minimal inhibitory concentrations of eight different antibiotics were determined using a microdilution plate. Similar to findings in European countries, the most common source of bacterial...... ICUs) or Enterococcus spp. and Klebsiella spp. (from hematology/oncology units), are the most prominent pathogens in these units today. Indicator organisms of antibiotic consumption (Pseudomonas aeruginosa and methicillin-resistant coagulase-negative staphylococci and Staphylococcus aureus) were more......The prevalence and antibiotic sensitivity patterns of bacteria collected consecutively from medical and surgical intensive care units (ICUs) and from hematology/oncology units in nine hospitals in Denmark were determined and compared to data collected simultaneously in 12 other European countries...

  9. Contamination profile for staphylococci and its enterotoxins and monitorization of the conditions of hygiene in a 'coalho' cheese production line

    OpenAIRE

    Borges, MD; Nassu, RT; PEREIRA, JL; de Andrade, APC; Kuaye, AY

    2008-01-01

    This research aimed to evaluate the contamination by staphylococci and its enterotoxins as well as to monitor the Conditions of hygiene from a coalho cheese production like, using ATP bioluminescence assay. Staphylococcus sp. population varied from < ICFU mL(-1), in pasteurized milk to 1.5 x 10(7)CFU mL(-1) in raw milk, whereas coagulase-positive staphylococci count ranged from < ICFU mL(-1), in pasteurized milk to 5.0 x 10(6)CFU mL(-1) in raw milk. Coagulase-positive staphylococci were detec...

  10. Evaluation of nitrofurantoin activity against the urinary isolates in the current scenario of antimicrobial resistance

    Directory of Open Access Journals (Sweden)

    Rajkumar Manojkumar Singh

    2015-01-01

    Full Text Available Introduction: Urinary tract infections (UTIs caused by antibiotic-resistant isolates have become a major health hazard in recent years as they are very difficult to treat, causing an increase in morbidity and mortality. Nitrofurantoin has been used successfully for decades for the prophylaxis and treatment of uncomplicated cystitis but the increased emergence of antibiotic resistance has made nitrofurantoin a suitable candidate for the treatment of UTI caused by multidrug-resistant pathogens. This study was taken up with the aim to determine the activity of nitrofurantoin against the wide range of resistant urinary gram-negative and gram-positive isolates. Materials and Methods: A total of 440 nonduplicate, gram-negative, and gram-positive uropathogens obtained between July 2013 and December 2014 from 3,780 fresh midstream urine samples were subjected to the VITEK-2 compact system for identification and antimicrobial susceptibility testing. Phenotypic methods for the detection of different beta-lactamases [extended spectrum beta-lactamases, AmpC beta-lactamases, metallo-beta-lactamases, and inhibitor-resistant beta-lactamases (IRT] and methicillin-resistance staphylococci [methicillin-resistant Staphylococcus aureus (MRSA and methicillin-resistant coagulase-negative staphylococci (MRCNS] were employed. Results: High activity of nitrofurantoin was observed against MRCNS (96% followed by MRSA (95%, extended spectrum beta-lactamases (ESBLs (70%, IRT (66%, and vancomycin-resistant enterococci (VRE (66%. However, the sensitivity rate was lowered to 38% and 32% for metallo-beta-lactamases (MBLs and AmpC beta-lactamases, respectively. Conclusion: In view of the current incidence of antibiotic resistance among community uropathogens and based on efficacy, cost-effectiveness, and low impact on promoting resistance, nitrofurantoin should be considered as a reasonable alternative to trimethoprim-sulfamethoxazole (TMP-SMX and fluoroquinolones for the first

  11. Selection of Strains for Quality Assessment of the Disk Induction Method for Detection of Inducible Clindamycin Resistance in Staphylococci: a CLSI Collaborative Study

    Science.gov (United States)

    Zelazny, Adrian M.; Ferraro, Mary Jane; Glennen, Anita; Hindler, Janet F.; Mann, Linda M.; Munro, Susan; Murray, Patrick R.; Reller, L. Barth; Tenover, Fred C.; Jorgensen, James H.

    2005-01-01

    A nine-laboratory collaborative study was conducted to select positive and negative quality assessment control strains for the detection of inducible clindamycin resistance in staphylococci. Four strains of Staphylococcus aureus were tested as unknowns on 10 different days in each laboratory using the recently recommended CLSI (formerly NCCLS) disk diffusion method and the inoculum purity control method. Strains contained either macrolide-lincosamide-streptogramin B (MLSB) resistance genes encoded by erm(A) or erm(C) or a macrolide resistance efflux pump encoded by msr(A). Based upon the results of this study, strain UT 32 (now designated ATCC strain BAA-977) containing erm(A) is recommended as the positive control organism for inducible clindamycin resistance. Strain UT 25 (now designated ATCC BAA-976), which harbors the efflux pump encoded by msr(A), is recommended as the negative control organism. PMID:15956373

  12. [Incidence of Kanagawa phenomenon-positive and -negative Vibrio parahaemolyticus strains isolated from traveller's diarrhea and their relation to tdh and trh genes].

    Science.gov (United States)

    Suzuki, N; Ueda, Y; Furukawa, T; Takegaki, Y; Miyagi, K; Noda, K; Hirose, H; Hashimoto, S; Yano, S; Ishibashi, M; Honda, T

    1997-05-01

    A total of 1,319 strains of Vibrio parahaemolyticus isolated from traveller's diarrhea were analysed for Kanagawa phenomenon (KP) with the Wagatsuma blood agar test and the results were also compared with those of analyses of tdh and trh genes which encode thermostable direct hemolysin (tdh) and TDH-related hemolysin (trh). The majority of the strains (1,152 strains) counting 87.3% had positive KP, among which 1,049 and 103 strains were only tdh and both tdh and trh-positive ones, respectively. However, 167 strains counting 12.7%, which is quite high compared to the previous report, were found to have negative KP, among which 94 and 24 strains were only trh and both tdh and trh-positive ones, respectively.

  13. In vitro susceptibility of characterized β-lactamase-producing Gram-negative bacteria isolated in Japan to ceftazidime-, ceftaroline-, and aztreonam-avibactam combinations.

    Science.gov (United States)

    Yoshizumi, Ayumi; Ishii, Yoshikazu; Aoki, Kotaro; Testa, Raymond; Nichols, Wright W; Tateda, Kazuhiro

    2015-02-01

    Avibactam displays potent inhibition of extended-spectrum, AmpC, KPC and some OXA β-lactamases. We examined the combinations of avibactam with ceftazidime, ceftaroline and aztreonam by the broth microdilution method against Gram-negative bacteria harboring molecularly-characterized β-lactamase genes collected in Toho University, Japan. Bacterial isolates included: Ambler class A β-lactamase-producing Enterobacteriaceae (n = 26); class C β-lactamase-producing Enterobacteriaceae (n = 9) and class D β-lactamase-producing Acinetobacter baumannii (n = 9) and Enterobacteriaceae (n = 3). Ceftazidime-avibactam, ceftaroline-avibactam ands aztreonam-avibactam were active against the strains with an extended-spectrum β-lactamase (ESBL) or AmpC enzymes, but combination with avibactam did not reduce β-lactam MICs against A. baumannii with OXA β-lactamases including carbapenemases, such as OXA-40 and -69.

  14. Characterization of coagulase-negative staphylococcus species from cows' milk and environment based on bap, icaA, and mecA genes and phenotypic susceptibility to antimicrobials and teat dips.

    Science.gov (United States)

    Piessens, V; De Vliegher, S; Verbist, B; Braem, G; Van Nuffel, A; De Vuyst, L; Heyndrickx, M; Van Coillie, E

    2012-12-01

    The aim of this study was to investigate whether the main coagulase-negative staphylococci (CNS) species involved in bovine intramammary infections (IMI) possess specific characteristics that promote colonization of the udder. Virulence markers associated with biofilm formation, antimicrobial resistance, and biocide tolerance were compared between typically contagious CNS species (Staphylococcus chromogenes, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus simulans) and those rarely causing IMI (Staphylococcus sciuri, Staphylococcus equorum, and others) to find possible associations with pathogenicity. Coagulase-negative staphylococci isolates (n=366) belonging to 22 different species were analyzed by PCR for the presence of the biofilm-associated genes bap and icaA, and the methicillin resistance gene mecA. A selection of 82 isolates was additionally tested for their susceptibility to 5 antibiotics and 2 commercial teat dip products. Minimum inhibitory concentrations of antimicrobials were determined by Etest (AB bioMérieux, Marcy l'Etoile, France), and a microdilution method was optimized to determine minimum biocidal concentrations of teat dips. The bap, icaA, and mecA genes were detected significantly more in isolates from CNS species typically living in the cows' environment than in isolates from IMI-causing species. Antimicrobial resistance was mainly against erythromycin (23%) or oxacillin (16%), and was detected more often in the environmental species. The isolates least susceptible to the teat dips belonged to the IMI-causing species Staph. chromogenes and Staph. simulans. We concluded that carriage of biofilm genes and antimicrobial resistance were not associated with the ability to colonize the mammary gland because free-living CNS species constituted a more significant reservoir of biofilm and resistance determinants than did IMI-causing species. In contrast, increased tolerance to biocides may favor the establishment of

  15. Cytosine deaminase as a negative selectable marker for the microalgal chloroplast: a strategy for the isolation of nuclear mutations that affect chloroplast gene expression.

    Science.gov (United States)

    Young, Rosanna E B; Purton, Saul

    2014-12-01

    Negative selectable markers are useful tools for forward-genetic screens aimed at identifying trans-acting factors that are required for expression of specific genes. Transgenic lines harbouring the marker fused to a gene element, such as a promoter, may be mutagenized to isolate loss-of-function mutants able to survive under selection. Such a strategy allows the molecular dissection of factors that are essential for expression of the gene. Expression of individual chloroplast genes in plants and algae typically requires one or more nuclear-encoded factors that act at the post-transcriptional level, often through interaction with the 5' UTR of the mRNA. To study such nuclear control further, we have developed the Escherichia coli cytosine deaminase gene codA as a conditional negative selectable marker for use in the model green alga Chlamydomonas reinhardtii. We show that a codon-optimized variant of codA with three amino acid substitutions confers sensitivity to 5-fluorocytosine (5-FC) when expressed in the chloroplast under the control of endogenous promoter/5' UTR elements from the photosynthetic genes psaA or petA. UV mutagenesis of the psaA transgenic line allowed recovery of 5-FC-resistant, photosynthetically deficient lines harbouring mutations in the nuclear gene for the factor TAA1 that is required for psaA translation. Similarly, the petA line was used to isolate mutants of the petA mRNA stability factor MCA1 and the translation factor TCA1. The codA marker may be used to identify critical residues in known nuclear factors and to aid the discovery of additional factors required for expression of chloroplast genes.

  16. Synergistic hemolytic reactions between staphylococci and Micrococcus lylae.

    Science.gov (United States)

    Lämmler, C; Brückler, J

    1989-06-01

    The primary culture of a clinical specimen obtained from a dog with an acute squamous eczema revealed three different bacterial species which demonstrated synergistic hemolytic activities on sheep blood agar plates. The three cultures were identified as beta-hemolytic Staphylococcus intermedius, as a coagulase-negative staphylococcal species, producing a delta-like hemolysin and as non-hemolytic Micrococcus lylae. The coagulase-negative staphylococcal species as well as M. lylae produced synergistically with beta-hemolytic S. intermedius zones of complete hemolysis. The occurrence of three different synergistically active bacterial species from one clinical specimen might be of clinical significance.

  17. Characterisation of a haemolysin related to Vp-TDH produced by a Kanagawa phenomenon-negative clinical isolate of Vibrio parahaemolyticus.

    Science.gov (United States)

    Nagayama, K; Yamamoto, K; Mitawani, T; Honda, T

    1995-02-01

    The production of a family of haemolysins--thermostable direct haemolysin (Vp-TDH), Vp-TDH-related haemolysin (Vp-TRH) and Vp-TDH/I--has been reported in clinical isolates of Vibrio parahaemolyticus. This paper describes a fourth type of haemolysin--Vp-TDH/II--produced by a Kanagawa phenomenon-negative clinical isolate of V. parahaemolyticus (O13:K, untypable). Vp-TDH/II was purified by ammonium sulphate precipitation and successive filtrations on DEAE-cellulose, hydroxyapatite, Sepharose 4B and Mono Q columns. Vp-TDH/II was biophysicochemically and immunologically similar to, but not identical to Vp-TDH, Vp-TRH and Vp-TDH/I. Vp-TDH/II stimulated vascular permeability in rabbit skin and was lethal to mice. Purified Vp-TDH/II and viable cells of the Vp-TDH/II-producing strain both induced fluid accumulation in ligated rabbit intestine. The plasmid-determined structural gene for Vp-TDH/II was cloned and the nucleotide sequence determined. The deduced amino acid sequence of Vp-TDH/II differed from those of Vp-TDH, Vp-TRH and Vp-TDH/I.

  18. Isolation and characterization of insertion sequence elements from gram-negative bacteria by using new broad-host-range, positive selection vectors.

    Science.gov (United States)

    Simon, R; Hötte, B; Klauke, B; Kosier, B

    1991-01-01

    On the basis of an RSF1010-derived broad-host-range vector, three different systems which enable positive detection and isolation of insertion sequence (IS) elements from gram-negative bacteria were constructed. Vectors pSUP104-pheS, pSUP104-rpsL, and pSUP104-sac were used successfully in a number of Rhizobium strains and in Xanthomonas campestris. More than 20 different IS elements were isolated and characterized. The 16 IS elements from Rhizobium meliloti were further used to characterize various R. meliloti strains by hybridization. The resulting hybridization patterns were different for every strain and gave a clear and definite IS fingerprint of each strain. These IS fingerprints can be used to identify and characterize R. meliloti strains rapidly and unequivocally, as they proved to be relatively stable. Some of the IS elements were found to be identical when the IS fingerprints from a given strain were compared. This method of IS fingerprinting can also establish whether IS elements are the same, related, or different. Images PMID:1847366

  19. Isolation and characterization of insertion sequence elements from gram-negative bacteria by using new broad-host-range, positive selection vectors.

    Science.gov (United States)

    Simon, R; Hötte, B; Klauke, B; Kosier, B

    1991-02-01

    On the basis of an RSF1010-derived broad-host-range vector, three different systems which enable positive detection and isolation of insertion sequence (IS) elements from gram-negative bacteria were constructed. Vectors pSUP104-pheS, pSUP104-rpsL, and pSUP104-sac were used successfully in a number of Rhizobium strains and in Xanthomonas campestris. More than 20 different IS elements were isolated and characterized. The 16 IS elements from Rhizobium meliloti were further used to characterize various R. meliloti strains by hybridization. The resulting hybridization patterns were different for every strain and gave a clear and definite IS fingerprint of each strain. These IS fingerprints can be used to identify and characterize R. meliloti strains rapidly and unequivocally, as they proved to be relatively stable. Some of the IS elements were found to be identical when the IS fingerprints from a given strain were compared. This method of IS fingerprinting can also establish whether IS elements are the same, related, or different.

  20. Comparative evaluation of Vitek 2 identification and susceptibility testing of Gram-negative rods directly and isolated from BacT/ALERT-positive blood culture bottles.

    Science.gov (United States)

    Munoz-Dávila, M J; Yagüe, G; Albert, M; García-Lucas, T

    2012-05-01

    The performance of Vitek 2 was evaluated for the identification and susceptibility testing of Gram-negative bacilli directly from positive blood cultures bottles. Direct inoculation of the positive blood cultures with the Vitek cards ID-GN and AST-NO58 was compared with the standard inoculation method based on the sub-culture of the positive blood culture to agar. A total of 142 blood cultures were included in the study; of those, 119 were from patients' clinical samples, while 23 were artificially prepared with strains showing different mechanisms of resistance. A total of 136 (95.8%) strains were correctly identified to the species level, only 2 (1.4%) were mis-identified and 4 (2.8%) were not identified. Susceptibility results were available for all isolates tested against 17 antibiotics, thus, resulting in a total of 2,414 isolate/anti-microbial combinations. The error rate was 2.8% (67/2,414) overall; 0.6% (14/2,414) very major errors, 0.1% (3/2,414) major errors and 2.1% (50/2,414) minor errors. The direct method detected 88.5% (22/25) of the strains producing extended-spectrum beta-lactamases (ESBLs). The susceptibility agreement among the added strains with ESBL, AMPc hyperproduction, resistance to ceftazidime, carbapenems and cefepime was very high. Direct identification and susceptibility testing gave rapid and reliable results, reducing by 24 h the turnaround time of the microbiology laboratory.

  1. Negative inotropic and dromotropic actions of SiO2 nanoparticles on isolated rat hearts: Effects on Na+ and Ca2+ currents

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    Julio Alvarez-Collazo

    2016-12-01

    Full Text Available Context: SiO2 nanoparticles (NP are widely used in the industry and in varied biotechnological and medical applications. However, epidemiological studies suggest that pollution with fine particles (in which silica is an inorganic component may increase morbidity and mortality from cardiovascular diseases, but little is known about their potential cardiovascular actions. Aims: To study the actions of SiO2 nanoparticles on the electrical and contractile activity of rat hearts and to identify the possible underlying cellular mechanisms. Methods: Surface electrogram (ECG and force of contraction (FC was recorded in isolated rat hearts. Na+ and Ca2+ currents (INa and ICaL, respectively were recorded, with the patch-clamp technique, in enzymatically isolated rat ventricular cardiomyocytes. Results: SiO2 NP (1-30 µg/mL decreased the FC and markedly increased QRS duration and QT interval in spontaneously beating hearts. Electric stimulation (RR = 400 ms partially restored the FC. In patch-clamp experiments NP (30 µg/mL decreased INa in a use-dependent manner and increased ICaL. Conclusions: SiO2 nanoparticles exert a negative inotropic action in rat hearts due, in part, to a decrease in the fast sodium current responsible for cardiac depolarization. SiO2 nanoparticles are also able to increase the L-type Ca2+ current. These actions should be taken into account when analyzing the toxic effects of these nanoparticles.

  2. National epidemiology of carbapenem-resistant and extensively drug-resistant Gram-negative bacteria isolated from blood samples in China in 2013.

    Science.gov (United States)

    Xu, A; Zheng, B; Xu, Y-C; Huang, Z-G; Zhong, N-S; Zhuo, C

    2016-03-01

    Although antimicrobial resistance poses a great challenge to clinicians in China, there are limited antimicrobial resistance data on Gram-negative bacteria nationwide. We investigated the phenotypic characteristics of carbapenem-resistant Escherichia coli (CREC) and Klebsiella pneumoniae (CRKP) as well as extensively drug-resistant strains of Pseudomonas aeruginosa (XDRPA) and Acinetobacter baumannii (XDRAB) isolated from blood cultures in China. Data were collected on 24113 isolates from the China surveillance of antimicrobial resistance program in 2013, which comprised 208 hospitals located in all seven administrative regions of China. Minimum inhibitory concentrations (MICs) for common antimicrobials were determined by commercial automated systems available at local hospitals, and associations with geographic and clinical distributions was further studied. The overall prevalence of CREC, CRKP, XDRAB and XDRPA strains was 1.0, 5.5, 13.7 and 4.2%, respectively. Except for CREC, which did not differ greatly by region, the prevalence of the remaining three strains varied significantly across regions. The highest prevalence of CRKP (10.6%) and XDRAB (13.1%) were found in the pediatric group, and higher prevalence of all four target strains was found in the intensive care unit. For imipenem, 55.8% of CREC and 22.9% of CRKP strains had MICs of ≤4 μg/mL, while 97.4% XDRAB and 84% XDRPA isolates had MICs of ≥16 μg/mL. All CREC, CRKP and 81.2% of XDRAB strains were susceptible to tigecycline, with MIC90 values of 0.5, 2 and 4 μg/mL, respectively. In conclusion, a high prevalence of CRKP and XDRAB has emerged in China, especially in children and in the intensive care unit.

  3. Prevalence of the Antibiotic Resistance Genes in Coagulase-Positive-and Negative-Staphylococcus in Chicken Meat Retailed to Consumers

    Science.gov (United States)

    Osman, Kamelia; Badr, Jihan; Al-Maary, Khalid S.; Moussa, Ihab M. I.; Hessain, Ashgan M.; Girah, Zeinab M. S. Amin; Abo-shama, Usama H.; Orabi, Ahmed; Saad, Aalaa

    2016-01-01

    The use of antibiotics in farm management (growing crops and raising animals) has become a major area of concern. Its implications is the consequent emergence of antibiotic resistant bacteria (ARB) and accordingly their access into the human food chain with passage of antibiotic resistance genes (ARG) to the normal human intestinal microbiota and hence to other pathogenic bacteria causative human disease. Therefore, we pursued in this study to unravel the frequency and the quinolone resistance determining region, mecA and cfr genes of methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA), methicillin-resistant coagulase-negative staphylococci (MRCNS) and methicillin-susceptible coagulase-negative staphylococci (MSCNS) isolated from the retail trade of ready-to-eat raw chicken meat samples collected during 1 year and sold across the Great Cairo area. The 50 Staphylococcus isolated from retail raw chicken meat were analyzed for their antibiotic resistance phenotypic profile on 12 antibiotics (penicillin, oxacillin, methicillin, ampicillin-sulbactam, erythromycin, tetracycline, clindamycin, gentamicin, ciprofloxacin, chloramphenicol, sulfamethoxazole-trimethoprim, and vancomycin) and their endorsement of the quinolone resistance determining region, mecA and cfr genes. The isolation results revealed 50 isolates, CPS (14) and CNS (36), representing ten species (S. aureus, S. hyicus, S. epidermedius, S. lugdunensis, S. haemolyticus, S. hominus, S. schleiferi, S. cohnii, S. intermedius, and S. lentus). Twenty seven isolates were methicillin-resistant. Out of the characterized 50 staphylococcal isolates, three were MRSA but only 2/3 carried the mecA gene. The ARG that bestows resistance to quinolones, β-lactams, macrolides, lincosamides, and streptogramin B [MLS(B)] in MRSA and MR-CNS were perceived. According to the available literature, the present investigation was a unique endeavor into the identification of the quinolone

  4. Prevalence of the antibiotic resistance genes in coagulase-positive- and negative-Staphylococcus in chicken meat retailed to consumers

    Directory of Open Access Journals (Sweden)

    Kamelia Mahmoud Osman

    2016-11-01

    Full Text Available The use of antibiotics in farm management (growing crops and raising animals has become a major area of concern. Its implications is the consequent emergence of antibiotic resistant bacteria (ARB and accordingly their access into the human food chain with passage of antibiotic resistance genes (ARG to the normal human intestinal microbiota and hence to other pathogenic bacteria causative human disease. Therefore, we pursued in this study to unravel the frequency and the quinolone resistance determining region, mecA and cfr genes of methicillin-susceptible Staphylococcus aureus (MSSA, methicillin-resistant S. aureus (MRSA, methicillin-resistant coagulase-negative staphylococci (MRCNS and methicillin-susceptible coagulase-negative staphylococci (MSCNS isolated from the retail trade of ready-to-eat raw chicken meat samples collected during one year and sold across the Great Cairo area. The 50 Staphylococcus isolated from retail raw chicken meat were analyzed for their antibiotic resistance phenotypic profile on 12 antibiotics (penicillin, oxacillin, methicillin, ampicillin-sulbactam, erythromycin, tetracycline, clindamycin, gentamicin, ciprofloxacin, chloramphenicol, sulfamethoxazole-trimethoprim and vancomycin and their endorsement of the quinolone resistance determining region, mecA and cfr genes. The isolation results revealed 50 isolates, CPS (14 and CNS (36, representing ten species (S. aureus, S. hyicus, S. epidermedius, S. lugdunensis, S. haemolyticus, S. hominus, S. schleiferi, S. cohnii, S. intermedius and S. lentus. Twenty seven isolates were methicillin-resistant. Out of the characterized 50 staphylococcal isolates, three were MRSA but only 2/3 carried the mecA gene. The ARG that bestows resistance to quinolones, β-lactams, macrolides, lincosamides and streptogramin B (MLS(B in MRSA and MR-CNS were perceived. According to the available literature, the present investigation was a unique endeavor into the identification of the quinolone

  5. Insights into the extremotolerance of Acinetobacter radioresistens 50v1, a gram-negative bacterium isolated from the Mars Odyssey spacecraft.

    Science.gov (United States)

    McCoy, K B; Derecho, I; Wong, T; Tran, H M; Huynh, T D; La Duc, M T; Venkateswaran, K; Mogul, R

    2012-09-01

    The microbiology of the spacecraft assembly process is of paramount importance to planetary exploration, as the biological contamination that can result from remote-enabled spacecraft carries the potential to impact both life-detection experiments and extraterrestrial evolution. Accordingly, insights into the mechanisms and range of extremotolerance of Acinetobacter radioresistens 50v1, a Gram-negative bacterium isolated from the surface of the preflight Mars Odyssey orbiter, were gained by using a combination of microbiological, enzymatic, and proteomic methods. In summary, A. radioresistens 50v1 displayed a remarkable range of survival against hydrogen peroxide and the sequential exposures of desiccation, vapor and plasma phase hydrogen peroxide, and ultraviolet irradiation. The survival is among the highest reported for non-spore-forming and Gram-negative bacteria and is based upon contributions from the enzyme-based degradation of H(2)O(2) (catalase and alkyl hydroperoxide reductase), energy management (ATP synthase and alcohol dehydrogenase), and modulation of the membrane composition. Together, the biochemical and survival features of A. radioresistens 50v1 support a potential persistence on Mars (given an unintended or planned surface landing of the Mars Odyssey orbiter), which in turn may compromise the scientific integrity of future life-detection missions.

  6. Fast DNA isolation and PCR protocols for detection of methicillin-resistant staphylococci.

    Science.gov (United States)

    Adwan, Kamel

    2014-01-01

    A rapid method that included simple boiling DNA extraction followed by a fast polymerase chain reaction (PCR) cycling protocol designed to detect mecA, which characterizes methicillin-resistant Staphylococcus aureus (MRSA), was performed. Briefly, the PCR cycling protocol consisted of pre-denaturation at 95 °C for 30 s, 30 cycles of denaturation at 94 °C for 2 s, annealing at 52 °C for 5 s, extension for 10 s, and final extension at 72 °C for 1 min. A good level of reliability of the method was verified. The study has shown that the method described here represents a rapid and accurate DNA extraction and PCR-based identification system of MRSA, thus allowing clinicians to make early identification and early implementation of control measures.

  7. Distribution of methicillin-resistant coagulase-positive staphylococci (MRCoPS) in a surgical unit and cystotomy operation sites in a veterinary teaching hospital.

    Science.gov (United States)

    Fungwithaya, Punpichaya; Brikshavana, Pasakorn; Chanchaithong, Pattrarat; Prapasarakul, Nuvee

    2017-02-28

    This study aimed to investigate the spread of methicillin-resistant coagulase-positive staphylococci (MRCoPS) among veterinary staff, hand-touch sites and surgical tissue during cystotomy operations on cats and dogs that were patients, and to analyze the genetic relatedness and antimicrobial resistance profiles of the isolates. Human and environmental samples were obtained from the nasal passageways of 12 surgeons and veterinary assistants and from 29 hand-touch sites of instruments in operative units and subjected to bacterial isolation and enumeration. Swab samples were collected in triplicate from 29 dogs and three cats at the site of incision, from the incision area, from the peritoneum during surgery and from the peritoneum before suture. MRCoPS were identified by mecA gene detection and characterized by their antibiogram profile, SCCmec type and pulsed-field gel electrophoresis. Twenty-four staphylococci were isolated, derived from one veterinary assistant, 12 operating room floor areas and hand-touch sites, three dogs and one cat. Methicillin-resistant S. pseudintermedius (MRSP) was found on an electric clipper and rebreathing circuits in the operating room. Three dogs were positive for MRSP during surgery, and one methicillin-resistant S. aureus (MRSA) was detected in a cat. All MRCoPS were resistant to doxycycline, erythromycin, clindamycin and enrofloxacin, but no patients developed surgical site infections. According to their genotypic patterns, the clones obtained from the environment and human sources differed from the animal clones. Despite intensive hygienic management, a variety of MRCoPS clones were present within the surgical unit and during surgery.

  8. Phenotypic and Molecular Antibiotic Resistance Determination of Airborne Coagulase Negative Staphylococcus spp. Strains from Healthcare Facilities in Southern Poland.

    Science.gov (United States)

    Lenart-Boroń, Anna; Wolny-Koładka, Katarzyna; Stec, Joanna; Kasprowic, Andrzej

    2016-10-01

    This study assessed the antimicrobial resistance of airborne Staphylococcus spp. strains isolated from healthcare facilities in southern Poland. A total of 55 isolates, belonging to 10 coagulase-negative staphylococci (CoNS) species, isolated from 10 healthcare facilities (including hospitals and outpatient units) were included in the analysis. The most frequently identified species were Staphylococcus saprophyticus and Staphylococcus warneri, which belong to normal human skin flora, but can also be the cause of common and even severe nosocomial infections. Disk diffusion tests showed that the bacterial strains were most frequently resistant to erythromycin and tetracycline and only 18% of strains were susceptible to all tested antimicrobials. Polymerase chain reaction amplification of specific gene regions was used to determine the presence of the Macrolide-Lincosamide-Streptogramin resistance mechanisms in CoNS. The molecular analysis, conducted using specific primer pairs, identified the msrA1 gene, encoding active efflux pumps in bacterial cells, as the most frequent resistance gene. As many as seven antibiotic resistance genes were found in one isolate, whereas the most common number of resistance genes per isolate was five (n = 17). It may be concluded that drug resistance was widely spread among the tested strains, but the resulting antimicrobial resistance profile indicates that in the case of infection, the use of antibiotics from the basic antibiogram group will be effective in therapy. However, before administering treatment, determination of the specific antimicrobial resistance should be conducted, particularly in the case of hospitalized patients.

  9. Comparative evaluation of the Vitek-2 Compact and Phoenix systems for rapid identification and antibiotic susceptibility testing directly from blood cultures of Gram-negative and Gram-positive isolates.

    Science.gov (United States)

    Gherardi, Giovanni; Angeletti, Silvia; Panitti, Miriam; Pompilio, Arianna; Di Bonaventura, Giovanni; Crea, Francesca; Avola, Alessandra; Fico, Laura; Palazzo, Carlo; Sapia, Genoveffa Francesca; Visaggio, Daniela; Dicuonzo, Giordano

    2012-01-01

    We performed a comparative evaluation of the Vitek-2 Compact and Phoenix systems for direct identification and antimicrobial susceptibility testing (AST) from positive blood culture bottles in comparison to the standard methods. Overall, 139 monomicrobial blood cultures, comprising 91 Gram-negative and 48 Gram-positive isolates, were studied. Altogether, 100% and 92.3% of the Gram-negative isolates and 75% and 43.75% of the Gram-positive isolates showed concordant identification between the direct and the standard methods with Vitek and Phoenix, respectively. AST categorical agreements of 98.7% and 99% in Gram-negative and of 96.2% and 99.5% in Gram-positive isolates with Vitek and Phoenix, respectively, were observed. In conclusion, direct inoculation procedures for Gram-negative isolates showed an excellent performance with both automated systems, while for identification of Gram-positive isolates they proved to be less reliable, although Vitek provided acceptable results. This approach contributes to reducing the turnaround time to result of blood cultures, with a positive impact on patient care.

  10. A small-scale, low-cost isolation system for the incubation and rearing of low bacterial load chicks as a model to study microbial-intestinal interactions.

    Science.gov (United States)

    Forder, Rebecca E A; Firth, Gordon A; Tivey, David R; Howarth, Gordon S; Hughes, Robert J

    2008-04-01

    A small-scale, economical isolator system was adapted to hatch and raise chicks in a bacteria-free environment as a means to observe bacterial interactions with the intestinal mucosa during early development. The design and construction of flexible plastic isolators for incubation and brooding are described along with methodologies for preparation of eggs for entry into the isolators, incubation and hatching. Two trials were conducted, the first in August 2005 and the second in March 2006. Results from both trials showed no differences in body weights of chicks raised in isolation when compared with those raised conventionally. Growth of bacteria was detected from rectal swabs at day 2 post-hatch, with both trials, showing a light growth of Bacillus sp., coagulase-negative staphylococci and haemolytic streptococcus in trial 1, and a light growth of Bacillus cereus only in trial 2. Although not germfree, the growth of bacteria in chicks raised in isolation was decreased or absent when compared with chicks raised conventionally. Feed was negative for contamination and surface swabs of equipment were also negative until day 3 post-hatch, suggesting possible contamination within the eggs themselves. Despite the presence of bacterial species, the isolator system was successful in producing low bacterial load chicks for comparison studies with conventionally raised chicks.

  11. Frequency of Blood Culture Isolates and their Antibiogram in a Teaching Hospital

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    Subha Shrestha

    2014-03-01

    Full Text Available Introduction: Bloodstream infections are associated with significant patient morbidity and mortality. Antimicrobial susceptibility patterns should guide the choice of empiric antimicrobial regimens for patients with bacteremia. Methods: Blood sample received from the patient attending Nepal Medical College and Teaching Hospital from March 2013 – August, 2013 were subjected to for culture. Isolate identification and antimicrobial susceptibility testing was done by standard microbiological method Results: Out of the total 2,766 blood samples, 13.3% showed bacterial growth. The percentage of neonatal septicemia was 13.3%. Staphylococcus aureus (28% was the most common isolates followed by Salmonella enterica Serotype Typhi (22%, Coagulase negative Staphylococci (9.5%, Salmonella enterica Serotype Paratyphi ((7.6% and Klebsiella pneumoniae (7.6%. 26.3% of the isolates of Staphylococcus aureus were oxacillin resistant. Most of the gram positive organisms were susceptible to amikacin and vancomycin and showed high level resistance to cefuroxime and cotrimoxazole. Out of 109 isolates of typhoid bacilli, 95.3% were resistant to nalidixic acid ,79% to ciprofloxacin and 60.5% to ofloxacin. More than 50% of the isolates of Klebsiella pneumoniae and Escherichia coli showed resistance to cephalosporins and cotrimoxazole. Acinetobacter spp showed high resistance (more than 60% to ceftriaxone and ofloxacin. More than 20% of the isolates of Pseudomonas aeruginosa were resistant to ciprofloxacin and amikacin. Conclusions: Ongoing surveillance for antimicrobial susceptibility remains essential, and will enhance efforts to identify resistance and attempt to limit its spread. Keywords: antibiotic; bacteria; blood stream infections.

  12. Trends of Bacterial Keratitis Culture Isolates in Jerusalem; a 13- Years Analysis

    Science.gov (United States)

    Politis, Michael; Wajnsztajn, Denise; Rosin, Boris; Block, Colin; Solomon, Abraham

    2016-01-01

    Purpose To describe the trends in pathogens and antibacterial resistance of corneal culture isolates in infectious keratitis during a period of 13 years at Hadassah-Hebrew University Medical Center. Methods A Retrospective analysis of bacterial corneal isolates was performed during the months of January 2002 to December 2014 at Hadassah Hebrew University Medical Center. Demographics, microbiological data and antibiotic resistance and sensitivity were collected. Results A total of 943 corneal isolates were analyzed during a 13 year period. A total of 415 positive bacterial cultures and 37 positive fungal cultures were recovered, representing 48% of the total cultures. The Annual incidence was 34.78 ± 6.54 cases. The most common isolate was coagulase-negative staphylococcus (32%), which had a significant decrease in trend throughout the study period (APC = -8.1, p = 0.002). Methicillin-resistant Staphylococcus aureus (MRSA) appears to have a decrease trend (APC = -31.2, P = 0.5). There was an increase in the resistance trend of coagulase-negative staphylococci to penicillin (APC = 5.0, P = keratitis. There was no significant change in the annual incidence of cases of bacterial keratitis seen over the past 13 years. Keratitis caused by MRSA appeared to decrease in contrast to the reported literature. PMID:27893743

  13. Staphylococcus capitis isolated from prosthetic joint infections.

    Science.gov (United States)

    Tevell, S; Hellmark, B; Nilsdotter-Augustinsson, Å; Söderquist, B

    2017-01-01

    Further knowledge about the clinical and microbiological characteristics of prosthetic joint infections (PJIs) caused by different coagulase-negative staphylococci (CoNS) may facilitate interpretation of microbiological findings and improve treatment algorithms. Staphylococcus capitis is a CoNS with documented potential for both human disease and nosocomial spread. As data on orthopaedic infections are scarce, our aim was to describe the clinical and microbiological characteristics of PJIs caused by S. capitis. This retrospective cohort study included three centres and 21 patients with significant growth of S. capitis during revision surgery for PJI between 2005 and 2014. Clinical data were extracted and further microbiological characterisation of the S. capitis isolates was performed. Multidrug-resistant (≥3 antibiotic groups) S. capitis was detected in 28.6 % of isolates, methicillin resistance in 38.1 % and fluoroquinolone resistance in 14.3 %; no isolates were rifampin-resistant. Heterogeneous glycopeptide-intermediate resistance was detected in 38.1 %. Biofilm-forming ability was common. All episodes were either early post-interventional or chronic, and there were no haematogenous infections. Ten patients experienced monomicrobial infections. Among patients available for evaluation, 86 % of chronic infections and 70 % of early post-interventional infections achieved clinical cure; 90 % of monomicrobial infections remained infection-free. Genetic fingerprinting with repetitive sequence-based polymerase chain reaction (rep-PCR; DiversiLab®) displayed clustering of isolates, suggesting that nosocomial spread might be present. Staphylococcus capitis has the potential to cause PJIs, with infection most likely being contracted during surgery or in the early postoperative period. As S. capitis might be an emerging nosocomial pathogen, surveillance of the prevalence of PJIs caused by S. capitis could be recommended.

  14. Diversity of Staphylococcus species and prevalence of enterotoxin genes isolated from milk of healthy cows and cows with subclinical mastitis.

    Science.gov (United States)

    Rall, V L M; Miranda, E S; Castilho, I G; Camargo, C H; Langoni, H; Guimarães, F F; Araújo Júnior, J P; Fernandes Júnior, A

    2014-02-01

    The objectives of this study were to determine the occurrence and diversity of Staphylococcus spp. in milk from healthy cows and cows with subclinical mastitis in Brazil and to examine the profile of enterotoxin genes and some enterotoxins produced by Staphylococcus spp. A total of 280 individual mammary quarter milk samples from 70 healthy cows and 292 samples from 73 cows with subclinical mastitis were collected from 11 farms in the state of São Paulo, Brazil. Staphylococcus spp. were recovered from 63 (22.5%) samples from healthy cows and from 80 samples (27.4%) from cows with mastitis. The presence of Staphylococcus aureus was significantly different between these 2 groups and was more prevalent in the cows with mastitis. The presence of Staphylococcus saprophyticus was also significantly different between these 2 groups, but this organism was more prevalent in healthy cows. No statistically significant differences were observed in the numbers of other staphylococci in milk samples from the 2 groups. The sea gene was the most prevalent enterotoxin gene in both groups. Eight of 15 (53.3%) Staph. aureus carried this gene and all produced the SEA toxin. In the coagulase-negative staphylococci (CNS) group, 61 of 128 (47.5%) had the same gene and just 1 (1.6%) Staphylococcus epidermidis strain produced the enterotoxin in vitro. Because CNS were isolated from both groups of cows and most CNS contained enterotoxin genes but did not produce toxins, the role of CNS in mastitis should be carefully defined.

  15. Prevalence and antibiotic susceptibility of coagulase-negative Staphylococcus species from bovine subclinical mastitis in dairy herds in the central region of Argentina.

    Science.gov (United States)

    Raspanti, Claudia G; Bonetto, Cesar C; Vissio, Claudina; Pellegrino, Matías S; Reinoso, Elina B; Dieser, Silvana A; Bogni, Cristina I; Larriestra, Alejandro J; Odierno, Liliana M

    2016-01-01

    Coagulase-negative staphylococci (CNS) are a common cause of bovine subclinical mastitis (SCM). The prevalence of CNS species causing SCM identified by genotyping varies among countries. Overall, the antimicrobial resistance in this group of organisms is increasing worldwide; however, little information exists about a CNS species resistant to antibiotics. The aim of the present study was to genotypically characterize CNS at species level and to determine the prevalence and antibiotic resistance profiles of CNS species isolated from bovine SCM in 51 dairy herds located in the central region of the province of Cordoba, Argentina. In this study, we identified 219 CNS isolates at species level by PCR-restriction fragment length polymorphism of the groEL gene. Staphylococcus chromogenes (46.6%) and Staphylococcus haemolyticus (32%) were the most prevalent species. A minimum of three different CNS species were present in 41.2% of the herds. S. chromogenes was isolated from most of the herds (86.3%), whereas S. haemolyticus was isolated from 66.7% of them. The broth microdilution method was used to test in vitro antimicrobial susceptibility. Resistance to a single compound or two related compounds was expressed in 43.8% of the isolates. S. chromogenes and S. haemolyticus showed a very high proportion of isolates resistant to penicillin. Resistance to two or more non-related antimicrobials was found in 30.6% of all CNS. S. haemolyticus exhibited a higher frequency of resistance to two or more non-related antimicrobials than S. chromogenes.

  16. Virulence factors genes of Staphylococcus spp. isolated from caprine subclinical mastitis.

    Science.gov (United States)

    Salaberry, Sandra Renata Sampaio; Saidenberg, André Becker Simões; Zuniga, Eveline; Melville, Priscilla Anne; Santos, Franklin Gerônimo Bispo; Guimarães, Ednaldo Carvalho; Gregori, Fábio; Benites, Nilson Roberti

    2015-08-01

    The aim of this study was to investigate genes involved in adhesion expression, biofilm formation, and enterotoxin production in isolates of Staphylococcus spp. from goats with subclinical mastitis and associate these results with the staphylococcal species. One hundred and twenty-four isolates were identified and polymerase chain reaction (PCR) was performed to detect the following genes: cna, ebpS, eno, fib, fnbA, fnbB, bap, sea, seb, sec, sed and see. The most commonly Staphylococcus species included S. epidermidis, S. lugdunensis, S. chromogenes, S. capitis ss capitis and S. intermedius. With the exception of fnbB, the genes were detected in different frequencies of occurrence in 86.3% of the Staphylococcus spp. isolates. Eno (73.2%) and bap (94.8%) were more frequently detected in coagulase-negative staphylococci (CNS); ebpS (76%), fib (90.9%) and fnbA (87%) were the most frequent genes in coagulase-positive staphylococci (CPS). Regarding enterotoxins, genes sed (28.2%) and see (24.2%) had a higher frequency of occurrence; sec gene was more frequently detected in CPS (58.8%). There was no association between the presence of the genes and the Staphylococcus species. Different virulence factors genes can be detected in caprine subclinical mastitis caused by CNS and CPS. The knowledge of the occurrence of these virulence factors is important for the development of effective control and prevention measures of subclinical mastitis caused by CNS and CPS in goats. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. In Vitro Activity of the Siderophore Cephalosporin, Cefiderocol, against a Recent Collection of Clinically Relevant Gram-Negative Bacilli from North America and Europe, Including Carbapenem-Nonsusceptible Isolates (SIDERO-WT-2014 Study).

    Science.gov (United States)

    Hackel, Meredith A; Tsuji, Masakatsu; Yamano, Yoshinori; Echols, Roger; Karlowsky, James A; Sahm, Daniel F

    2017-09-01

    Cefiderocol (formerly S-649266) is an investigational siderophore cephalosporin. Iron-depleted cation-adjusted Mueller-Hinton broth (ID-CAMHB) was prepared according to the Clinical and Laboratory Standards Institute (CLSI) protocol and used to perform broth microdilution testing of cefiderocol against a 2014-2015 collection of clinical isolates of Gram-negative bacilli from North America (n = 4,239) and Europe (n = 4,966). The concentrations of cefiderocol inhibiting 90% of isolates tested (MIC90s) were 0.5 μg/ml (North America; n = 3,007) and 1 μg/ml (Europe; n = 3,080) for all isolates of Enterobacteriaceae; 1 μg/ml (North America; n = 30) and 4 μg/ml (Europe; n = 139) for meropenem-nonsusceptible (MIC ≥ 2 μg/ml) isolates of Enterobacteriaceae; 0.5 μg/ml for both North American (n = 765) and European (n = 765) isolates of Pseudomonas aeruginosa; 0.5 μg/ml (North America; n = 151) and 1 μg/ml (Europe; n = 202) for meropenem-nonsusceptible (MIC ≥ 4 μg/ml) isolates of P. aeruginosa; 1 μg/ml for both North American (n = 309) and European (n = 839) isolates of all Acinetobacter baumannii strains as well as for both North American (n = 173) and European (n = 595) isolates of meropenem-nonsusceptible A. baumannii; and 0.5μg/ml (North America; n = 152) and 0.25 μg/ml (Europe; n = 276) for isolates of Stenotrophomonas maltophilia MICs of cefiderocol were ≤4 μg/ml for 99.9% (6,078/6,087) of all Enterobacteriaceae, 97.0% (164/169) of meropenem-nonsusceptible Enterobacteriaceae, 99.9% (1,529/1,530) of all P. aeruginosa isolates, 100% (353/353) of meropenem-nonsusceptible P. aeruginosa isolates, 97.6% (1,120/1,148) of all A. baumannii isolates, 96.9% (744/768) of meropenem-nonsusceptible A. baumannii isolates, 100% of isolates of S. maltophilia (428/428) and 93.8% of isolates of Burkholderia cepecia (11/12). We conclude that cefiderocol demonstrated potent in vitro activity against a recent collection of clinical isolates of commonly encountered Gram-negative

  18. Clinical management issues of coagulase-negative staphylococcal sepsis in the neonatal intensive care unit

    NARCIS (Netherlands)

    Hemels, M.A.C.

    2011-01-01

    Nosocomial sepsis is a major cause of morbidity in the neonatal intensive care unit (NICU), with coagulase-negative staphylococci (CONS) generally reported to be the most frequent causative micro-organisms. There is substantial evidence for the association between CONS sepsis and indwelling intravas

  19. Isolation and characterization of a metastatic hybrid cell line generated by ER negative and ER positive breast cancer cells in mouse bone marrow.

    Directory of Open Access Journals (Sweden)

    Keya De Mukhopadhyay

    Full Text Available BACKGROUND: The origin and the contribution of breast tumor heterogeneity to its progression are not clear. We investigated the effect of a growing orthotopic tumor formed by an aggressive estrogen receptor (ER-negative breast cancer cell line on the metastatic potential of a less aggressive ER-positive breast cancer cell line for the elucidation of how the presence of heterogeneous cancer cells might affect each other's metastatic behavior. METHODS: ER positive ZR-75-1/GFP/puro cells, resistant to puromycin and non-tumorigenic/non-metastatic without exogenous estrogen supplementation, were injected intracardiacally into mice bearing growing orthotopic tumors, formed by ER negative MDA-MB-231/GFP/Neo cells resistant to G418. A variant cell line B6, containing both estrogen-dependent and -independent cells, were isolated from GFP expressing cells in the bone marrow and re-inoculated in nude mice to generate an estrogen-independent cell line B6TC. RESULTS: The presence of ER negative orthotopic tumors resulted in bone metastasis of ZR-75-1 without estrogen supplementation. The newly established B6TC cell line was tumorigenic without estrogen supplementation and resistant to both puromycin and G418 suggesting its origin from the fusion of MDA-MB-231/GFP/Neo and ZR-75-1/GFP/puro in the mouse bone marrow. Compared to parental cells, B6TC cells were more metastatic to lung and bone after intracardiac inoculation. More significantly, B6TC mice also developed brain metastasis, which was not observed in the MDA-MB-231/GFP/Neo cell-inoculated mice. Low expression of ERα and CD24, and high expression of EMT-related markers such as Vimentin, CXCR4, and Integrin-β1 along with high CD44 and ALDH expression indicated stem cell-like characteristics of B6TC. Gene microarray analysis demonstrated a significantly different gene expression profile of B6TC in comparison to those of parental cell lines. CONCLUSIONS: Spontaneous generation of the novel hybrid cell

  20. In vitro activity of Melaleuca alternifolia (tea tree) oil against bacterial and Candida spp. isolates from clinical specimens.

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    Banes-Marshall, L; Cawley, P; Phillips, C A

    2001-01-01

    This study investigates the in vitro activity of tea tree oil (TTO) against a range of wild strains of microorganisms isolated from clinical specimens of leg ulcers and pressure sores. The antimicrobial effectiveness of TTO is determined in terms of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) or minimum fungicidal concentration (MFC). The isolates include methicillin-resistant Staphylococcus aureus (MRSA), S. aureus, faecal streptococci, beta-haemolytic streptococci, coagulase-negative staphylococci, Pseudomonas spp. and coliform bacilli. Eleven Candida spp. isolates from skin and vaginal swabs also are tested. Using an agar dilution assay, the MICs of TTO in 88 out of 90 isolates was 0.5-1.0% (v/v), whilst with P. aeruginosa it was >2% (v/v). A broth microdilution method was used to determine MIC and minimum cidal concentration (MCC) of 80 isolates. In 64 isolates, TTO produced an inhibitory and cidal effect at 3% and 4% (v/v), respectively. S. aureus and Candida spp. were the most susceptible to TTO, with MICs and MBCs of 0.5% and 1%, respectively. P. aeruginosa and the faecal streptococci isolates, with MICs and MBCs of >8%, were resistant to TTO.

  1. Investigation of biofilm forming ability in Staphylococci causing bovine mastitis using phenotypic and genotypic assays.

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    Darwish, Samah F; Asfour, Hanaa A E

    2013-01-01

    A total of 40 S. aureus and 68 coagulase negative Staphylococcus (CNS) isolates from bovine subclinical mastitis were investigated for their ability to form biofilm as one of the most important virulence factors.Using Congo Red Agar (CRA) method, 32.5%, 35%, and 32.5% of S. aureus strains were strong, intermediate, and negative biofilm producers, while in CNS the percentages were 29.5%, 42.6%, and 27.9%, respectively. By microtiter plate (MTP) method, 52.5%, 27.5%, and 20% of S. aureus isolates were strong, moderate, and weak biofilm producers, while in CNS the percentages were 44%, 30.9%, and 19.2%, respectively. Indian ink staining was used to detect the EPS layer of biofilm producers. All isolates were screened for presence of biofilm related genes, eno, icaA, icaD, and bap. In S. aureus isolates, the positive rates of eno, icaA, icaD, and bap genes were 75%, 15%, 62.5%, and 2.5% while in CNS were 92.6%, 5.9%, 47.1%, and 4.4%, respectively. The eno gene had the highest rate while the bap gene had the lowest rate. Presence of icaA and icaD genes was not always correlated with biofilm production. This study demonstrated high prevalence of Staphylococcus biofilm producers among bovine mastitis in Egypt. Therefore, attention must be paid toward implementation of new ways for effective treatment of such infections.

  2. Evaluation of a simple protein extraction method for species identification of clinically relevant staphylococci by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

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    Matsuda, Naoto; Matsuda, Mari; Notake, Shigeyuki; Yokokawa, Hirohide; Kawamura, Yoshiaki; Hiramatsu, Keiichi; Kikuchi, Ken

    2012-12-01

    In clinical microbiology, bacterial identification is labor-intensive and time-consuming. A solution for this problem is the use of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). In this study, we evaluated a modified protein extraction method of identification performed on target plates (on-plate extraction method) with MALDI-TOF (Bruker Microflex LT with Biotyper version 3.0) and compared it to 2 previously described methods: the direct colony method and a standard protein extraction method (standard extraction method). We evaluated the species of 273 clinical strains and 14 reference strains of staphylococci. All isolates were characterized using the superoxide dismutase A sequence as a reference. For the species identification, the on-plate, standard extraction, and direct colony methods identified 257 isolates (89.5%), 232 isolates (80.8%), and 173 isolates (60.2%), respectively, with statistically significant differences among the three methods (P extraction method is at least as good as standard extraction in identification rate and has the advantage of a shorter processing time.

  3. Distribution and Antimicrobial Susceptibility Pattern of Gram Negative Bacteria Causing Urinary Tract Infection (UTI and Detection New Delhi Metallo-beta-lactamase-1 (NDM-1 Producing Isolates in Ahwaz

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    Parviz Afrugh

    2016-04-01

    Full Text Available Background: Urinary tract infection (UTI is the commonest bacterial infectious disease in worldwide (especially in developing countries with a high rate of morbidity and financial cost. The management of UTI infections has been jeopardized by increase in immergence of antimicrobial drug resistance. Knowledge of the local bacterial etiology and susceptibility patterns is required to trace any change that might have occurred in time so that updated recommendation for optimal empirical therapy of UTI can be made. The aim of this investigation was distribution and antimicrobial susceptibility pattern of gram negative bacteria causing urinary tract infection (UTI and detection NDM-1 (new-delhi-metallo-beta-lactamase-1 producing isolates in Ahwaz. Materials and Methods: This cross-sectional study was done during a period of one year from April 2013 to March 2014. Clean catch midstream urine samples were collected from suspected patients to UTI. The isolates were identified based on morphological and biochemical testes. Culture was performed on routine microbiological media. Susceptibility testing was performed according CLSI (2013 guidelines. Detection of carbapenemase producing isolates was performed by modified hodge test (MHT. Metallo-beta-lactamase isolates were detected by imipenem-EDTA combined disc test (CDT. Results: In this study 708 gram negative organisms were isolated from urine samples. E.coli was the most common isolated bacteria (67% followed by Klebsiella spp. (26.5% and Enterobacter spp. (2.5%. In antibiotic susceptibility testing more than 90% of isolates were sensitive to tetracycline, ceftazidime, meropenem, amikacin, cefotaxime, imipenem, and cefepime. Isolates were more resistant to cephalothin (32%, co-trimoxazol (30.5%, and nalidixic acid (25%. Conclusion: In our results isolated organisms from outpatients showed very high sensitivity to common antibiotics. Continuous and regular monitoring of susceptibility pattern of

  4. Survey of marbofloxacin susceptibility of bacteria isolated from cattle with respiratory disease and mastitis in Europe.

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    Kroemer, S; Galland, D; Guérin-Faublée, V; Giboin, H; Woehrlé-Fontaine, F

    2012-01-01

    A monitoring programme conducted in Europe since 1994 to survey the marbofloxacin susceptibility of bacterial pathogens isolated from cattle has established the susceptibility of bacterial strains isolated before any antibiotic treatment from bovine mastitis and bovine respiratory disease (BRD) cases between 2002 and 2008. Minimum inhibitory concentration (MIC) was determined by a standardised microdilution technique. For respiratory pathogens, Pasteurella multocida and Mannheimia haemolytica isolates (751 and 514 strains, respectively) were highly susceptible to marbofloxacin (MIC≤0.03 µg/ml for 77.39 per cent of the strains) and only 1.75 per cent of M haemolytica strains were resistant (MIC≥4 µg/ml). Histophilus somni isolates (73 strains) were highly susceptible to marbofloxacin (0.008 to 0.06 µg/ml). Mycoplasma bovis MIC (171 strains) ranged from 0.5 to 4 µg/ml. For mastitis pathogens, the majority of Escherichia coli isolates were highly susceptible to marbofloxacin (95.8 per cent of 617 strains). Staphylococcus aureus and coagulase-negative staphylococci (568 and 280 strains) had a homogenous population with MIC centred on 0.25 µg/ml. Streptococcus uberis and Streptococcus dysgalactiae (660 and 217 strains) were moderately susceptible with MIC centred on 1 µg/ml. Marbofloxacin MIC for these various pathogens appeared stable over the seven years of the monitoring programme and was similar to previously published MIC results.

  5. Characterization of the staphylococcal cassette chromosome composite island of Staphylococcus haemolyticus SH32, a methicillin-resistant clinical isolate from China.

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    Dongliang Yu

    Full Text Available Staphylococcal cassette chromosome (SCC elements contribute considerably to virulence and resistance to antibiotic agents in staphylococci. SCC elements in coagulase-negative staphylococci (CoNS are highly diverse and there is evidence suggesting that they serve as a reservoir for antibiotic resistance genes in methicillin-resistant Staphylococcus aureus (MRSA. However, only a small number of SCC elements have been characterized in CoNS and their exact roles in the emergence and evolution of MRSA remain to be demonstrated. Here, we determined the structure of an SCC composite island (CISH32 found in the clinical Staphylococcus haemolyticus isolate SH32 by whole-genome DNA sequencing. CISH32 was 48 kb in length and mainly composed of two imperfect SCC elements, namely (i a ΨSCCmec(SH32 part containing a class C1 mec gene complex but lacking ccr genes and (ii a SCCSH32 part with a ccrA5B3 gene complex but lacking mec genes. In addition, CISH32 contained a type III restriction-modification system and several resistance loci, for example genes conferring resistance to cadmium and arsenic. ΨSCCmec(SH32 is almost entirely identical to a pseudo SCCmec element found in S. haemolyticus WCH1 and shares pronounced sequence similarity to a ΨSCCmec element of S. haemolyticus JCSC1435. However, staphyloco