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.

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.

Identification of coagulase-negative staphylococci with the API staph system.

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Gemmell, C G; Dawson, J E

1982-11-01

A kit for the identification of staphylococci based on the biochemical criteria proposed by Kloos and Schleifer (W.E. Kloos and K.H. Schleifer, J. Clin. Microbiol., 1:82-88, 1975) is now available commercially. The system was used to identify 100 strains of coagulase-negative staphylococci isolated from various body sites as the primary etiological agent of clinical infection. The increasing importance of staphylococci and their resistance to antibiotics provided the rationale for such an investigation. Over 90% of the Staphylococcus isolates were easily identified as to their species on the basis of their reaction profile to 19 biochemical tests included in the kit. The remainder, which showed minor variations, could also be assigned to the various species. Identification of the isolates was as follows: S. epidermidis, 54; S. haemolyticus, 5; S. simulans, 2; S. hominis, 1; S. capitis, 4; S. cohnii, 2; S. warneri, 2; S. xylosus, 8; and S. saprophyticus, 22. Antibiotic sensitivity patterns were determined for each of the isolates. Novobiocin resistance was detected in strains of S. saprophyticus and S. xylosus, a property hitherto recognized in Micrococcus sp. type 3 causing bacteriuria in young women. Resistance to penicillin was widespread among strains of several species, whereas resistance to tetracycline was mainly confined to strains of S. epidermidis. General resistance to sulfamethoxazole and nalidixic acid was found among all strains, with almost uniform sensitivity to the other drugs tested.

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.

Speciation and antibiotic susceptibility patterns of coagulase-negative staphylococci.

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Smith, D J; Kaplan, R L; Landau, W; Trenholme, G M

1982-08-01

During a six month period, 191 isolates of coagulase-negative staphylococci from blood, urine, cerebrospinal fluid and heart valves were identified to species level and tested for antimicrobial susceptibility. Seventy-one percent of isolates were Staphylococcus epidermidis, 8% Staphylococcus warneri, 7% Staphylococcus hominis, 7% Staphylococcus haemolyticus, 4% Staphylococcus capitis, 2% Staphylococcus saprophyticus and 1% Staphylococcus cohnii. Approximately 4% of isolates were felt to be associated with infection. Overall, 18% of isolates were susceptible to penicillin G, 61% oxacillin, 98% cephalothin, 98% cefamandole, 72% cefotaxime, 95% cefsulodin, 76% gentamicin, 64% clindamycin and 98% rifampicin. All isolates were susceptible to vancomycin. Vancomycin, rifampicin, cephalothin and cefamandole showed excellent activity against oxacillin-resistant isolates. With one exception, speciation was not helpful in determining whether or not an isolate was associated with infection.

Identification and detection of methicillin resistance in Non-Epidermidis coagulase-negative staphylococci

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Carina Secchi

Full Text Available The NCCLS (2004 presented a new methodology to detect, by disk-diffusion agar, oxacillin-resistance using a cefoxitin disk. We identified coagulase-negative staphylococci (SCoN to the species level and compared the use of cefoxitin disks (30 µg with oxacillin disks (1 µg, agar dilution (minimum inhibitory concentration of oxacillin and mecA gene detection in isolates of coagulase-negative bacteria other than Staphylococcus epidermidis (SCoNne. A total of 238 SCoNne was evaluated; oxacillin-resistance (the mecA gene was detected in 71% of the isolates. All methods gave 100% sensitivity, based on presence of the mecA gene. The specificity of the cefoxitin disk was 100%, while the oxacillin disk gave a specificity of 91% and agar dilution oxacillin gave a specificity of 88%. We conclude that the cefoxitin disk is an efficient test, and it is an easy method for use in clinical laboratories to detect oxacillin-resistance in staphylococci.

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

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.

Genotypic Characterization of Coagulase-negative Staphylococci Isolated from Sheep Milk in Slovakia

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Ivana Pilipčincová

2010-01-01

Full Text Available Hitherto very few reports are available presenting identification and molecular characterization of the coagulase negative staphylococci (CNS from sheep milk in the subclinical stage of mastitis. Furthermore, very scanty data are available on the epidemiological status of CNS in different Slovak provinces. Milk samples from 54 sheep farms located in eastern Slovak region were screened. A total 240 CNS were identified with series of biochemical testes (STAPH-API and subjected further for genotyping with the help of pulse field gel electrophoresis (PFGE. The most frequently occurring CNS species according the biochemical characterization were: S. epidermidis (36.3 %, S. caprae (21.3 %, S. hominis (6.6 %, S. chromogenes (6.3 %, S. xylosus (5.8 %, S. warneri (5.0 % and S. capitis (4.6 %. Further PFGE-based characterization of these isolates revealed six pulsotypes of the S. epidermidis, two of S. caprae, three of S. chromogenes, nine of S. hominis, five of S. capitis and seven of S. xylosus. These results contribute to knowledge of the epidemiological situation of the CNS from the subclinical form of mastitis in Slovakia.

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.

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

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Beytullah Kenar*, Yahya Kuyucuoğlu and Esra Şeker

2012-01-01

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...

Investigation into the prevalence, persistence and antibiotic resistance profiles of staphylococci isolated from euro currency.

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Gabriel, E M; Coffey, A; O'Mahony, J M

2013-08-01

The study set out to sample € 10 banknotes for the presence of coagulase-positive staphylococci (CPS) such as Staphylococcus aureus (S. aureus) and coagulase-negative staphylococci (CoNS) in Southern Ireland, to assess the levels of antibiotic resistance among those isolated, and determine the persistence of S. aureus on € 10 banknotes and € 2 coins. We report that 97% of € 10 banknotes screened (n = 155) harboured multiple species of staphylococci. From the generated bank of strains, a total of 150 representative staphylococci isolates were used for further study, 71 were CPS and 79 were CoNS. Of these, we found that 62% of the staphylococci demonstrated resistance to at least one of the first-line antibiotics (52.11% of CPS isolates and 76.71% of the CoNS isolates). Resistance to multiple antibiotics was seen in 31.18% of the resistant isolates. In relation to persistence studies, S. aureus was shown to remain viable on euro banknotes and coins for significant periods (on average, 19.33 days on € 10 banknotes and 16.67 days on € 2 coins) as determined using bioluminescence. We advocate the expansion of antibiotic surveillance programs, with a view to tracking/monitoring antibiotic resistance dissemination among environmental contaminants. Additionally, we propose that 'cashless transactions' should be encouraged in high-risk environments such as hospitals and healthcare settings, as well as stricter infection controls. Although it is accepted that circulating currency has the potential to harbour disease-causing pathogens, studies investigating prevalence and persistence of such pathogens on euro currency are virtually nonexistent. In an attempt to rectify this, we examined the prevalence of staphylococci on € 10 banknotes in Ireland and reported relatively high levels of antibiotic resistance among the isolates. Furthermore, we have established the persistence of S. aureus on euro currency for the first time. © 2013 The Society for Applied

cfr-mediated linezolid-resistant clinical isolates of methicillin-resistant coagulase-negative staphylococci from China.

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Song, Yunjia; Lv, Yuan; Cui, Lanqing; Li, Yun; Ke, Qian; Zhao, Yixuan

2017-03-01

Three linezolid-resistant coagulase-negative staphylococci (LR-CoNS), including two Staphylococcus cohnii and one Staphylococcus capitis, were isolated from 1104 clinical staphylococcal isolates across China in 2013-2014. Antibiotic susceptibilities of the bacteria were determined by the agar dilution method. PCR and DNA sequencing were performed to determine the potential molecular mechanism of linezolid resistance. The two linezolid-resistant S. cohnii isolates were subjected to pulsed-field gel electrophoresis (PFGE) to investigate their genetic relatedness. Primer walking, S1 nuclease PFGE and Southern blot hybridisation were conducted to ascertain the location and environment of the cfr gene. All three isolates were positive for the cfr gene. Amino acid mutations S158F and S158Y in the ribosomal protein L3 were identified in S. cohnii 13B289 and 13L105, respectively, both of which also had an additional substitution (D159Y) in L3. PFGE indicated that the two S. cohnii isolates belonged to diverse clonal strains. S1 nuclease PFGE and Southern blotting experiments indicated that the cfr gene of the three isolates resided on plasmids of similar size (ca. 35.4kb). The cfr-harbouring segments of S. capitis 13G350 and S. cohnii 13L105 were identical to plasmid pSS-01 reported previously. The cfr-carrying fragment of S. cohnii 13B289 was indistinguishable from the formerly described plasmid pSS-02. In conclusion, the presence of the cfr gene located on a plasmid was the main mechanism contributing to resistance to linezolid in the three staphylococcal isolates. Hence, timely detection and judicious use of antibiotics are essential to prevent further transmission of this resistance mechanism. Copyright © 2016 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.

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

Clinical and molecular features of methicillin-resistant,coagulase-negative staphylococci of pets and horses

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Kern, Andrea; Perreten, Vincent

2017-01-01

Objectives To determine the antibiotic resistance and fingerprint profiles of methicillin-resistant coagulase-negative staphylococci (MRCoNS) from animal infections among different practices and examine the history of antibiotic treatment. Methods Isolates were identified by mass spectrometry and tested for antimicrobial resistance by broth dilution, microarrays and sequence analysis of the topoisomerases. Diversity was assessed by PFGE, icaA PCR and staphylococcal cassette chromosome mec (SC...

The role of staphylococci in subclinical mastitis of cows and lytic phage isolation against to Staphylococcus aureus

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Aliye Gulmez Saglam

2017-12-01

Full Text Available Aim: This study was conducted to determine the role of Staphylococcus in the formation of subclinical mastitis in cows and to isolate the phage against isolated Staphylococcus aureus strains. Materials and Methods: In this study, 400 milk cows were screened by California Mastitis Test (CMT for subclinical mastitis and 235 udders of 96 cows, which were determined to be positive, were evaluated for Staphylococcus. Milk samples were evaluated using conventional and molecular methods. In addition, phage isolation studies were performed against S. aureus strains causing mastitis. Results: At the result of cultural examination, of 235 milk samples that were found as positive for mastitis by CMT, a total of 117 (49.7% Staphylococcus spp. were isolated as a distribution of 74 (63.24% coagulase-positive staphylococci and 43 (36.75% coagulase-negative staphylococci. Of these isolates, 76 (64.95% were characterized as S. aureus both conventional and molecular techniques. Lytic bacteriophages against two S. aureus strains which were isolated from mastitic milk samples were obtained from wastewater samples. Conclusion: The results of this study show that a significant portion of subclinical mastitis was formed by staphylococci. In addition, phage isolation against S. aureus strains isolated can be considered as one of the steps to be applied in the prophylaxis and treatment of such infections.

Fusidic acid resistance among staphylococci strains isolated from clinical specimens

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Özcan Deveci

2012-03-01

Full Text Available Objectives: The aim of this study was to investigate in vitrosusceptibility of fusidic acid to clinic isolates of staphylococci.Materials and methods: The forty-one coagulase negativestaphylococci (CNS and 18 Staphylococcus aureusstrains isolated from various clinical specimens were includedin this study. Staphylococci isolates were identifiedby conventional methods such as colony morphologyonto medium, gram staining, catalase and coagulasetests. According to “Clinical and Laboratory Standards Institute(CLSI” criteria, antimicrobial susceptibility testingof isolates was performed by Kirby-Bauer’s disk diffusionmethod.Results: The seventy-two percent of the isolated S.aureuswere defined as methicillin sensitive-S.aureus (MSSA,28% of the isolated S.aureus were defined as methicillinresistant-S.aureus (MRSA. The difference among fusidicacid susceptibility rates of MSSA and MRSA strains wasnot statistically significant (p=0.305. The twenty-nine percentof the isolated CNS were defined as methicillin sensitive-CNS (MS-CNS, 71% of the isolated CNS were definedas methicillin resistant-CNS (MR-CNS. There wasno statistically significant difference between MS-CNSand MR-CNS strains for fusidic acid susceptibility rates(p=0.490. But the difference among fusidic acid susceptibilityrates of CNS and S.aureus strains was statisticallysignificant (p<0.001. CNS strains were found more resistancethan S.aureus strains for fusidic acid.Conclusion: In this study, the resistance rates weredetected to increase for fusidic acid along with methicillinresistance. Among CNS isolates, fusidic acid resistancerates were significantly more elevated than that forS.aureus. Fusidic acid remains as an alternative in thetreatment of infections due to staphylococci.

Susceptibility of coagulase positive staphylococci isolated from cow's mammary gland to antibacterial drugs

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Savić-Rajić Nataša

2009-01-01

Full Text Available Coagulase positive staphylococci are one of the most common causes of chronic udder infection. Indiscriminate use of antimicrobial drugs and their presence in the environment where animals live has led to coagulase positive staphylococci strains resistant to antimicrobial means. Proper and timely treatment of sub-clinical mastitis, based on the most effective use of antimicrobial drugs, is the key to good health of the milk herd. The aim was to determine the antimicrobial efficacy of selected assets in relation to coagulase positive staphylococci isolated from samples of milk taken from individual udder quarters of cows in cases of udder infection from three farms with different mastitis prevalence. From a total of 9245 samples of milk taken from individual udder quarters of cows from three farms, 852 strains isolated were coagulase positive staphylococci. Coagulase positive staphylococci were isolated on blood agar and identified on the basis of macro-morphological characteristics and the coagulase and catalase test. The sensitivity of the coagulase positive staphylococci was tested by the Kirby Bauer agar diffusion method with the following antimicrobials: penicillin 6µg, amoxicillin / sulbactam (20 +10µg, cloxacillin 25 µg, cefalexin 30 µg, ceftiofur 30µg, linkomycin 15µg, 30 µg gentamycin and tetracycline 30 µg. Sensitivity testing of coagulase positive staphylococci, isolated in cases of intramammary cow infections, established a high degree of sensitivity in vitro towards penicilinasa resistant drugs (amoxicillin-sublactam, cloxacilin, cephalosporins of the first and third generations and linkomycin. The highest levels of resistance to penicillin (70.4% were found on a farm with a moderate prevalence of udder infection, then on the farm with the highest prevalence of intramammary infections (60.2% and the lowest on the farm with controlled levels of resistance of infection (43.7%. .

Coagulase-negative staphylococci mastitis in Dutch dairy herds

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Sampimon, O.C.

2009-01-01

Mastitis is one of the most important diseases in dairy cattle. Recently, the so-called minor pathogens, of which coagulase-negative staphylococci (CNS) are the most important group of bacteria, has received more attention. This thesis focuses on the role of CNS in udder health of dairy cows. The

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

DETECTION OF BIOFILM PRODUCTION IN BLOOD CULTURE ISOLATES OF STAPHYLOCOCCI

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Gupta Puja, Gupta Pratima, Mittal Garima, Agarwal RK, Goyal Rohit

2015-01-01

Full Text Available Background: Biofilm producing bacteria which are inherently resistant to antibiotics and disinfectants are widely associated with implant associated infections. Staphylococcus is the most commonly associated pathogens with bloodstream infection. Aims: The current study was conducted to detect biofilm production in Staphylococci isolated from blood culture specimens. Materials and Methods: 70 clinically significant staphylococcal isolates from blood culture were screened for biofilm production by Tissue culture plate (TCP method, Tube method (TM and Congo red agar (CRA method and their antibiotic susceptibility profile was studied. Results: 59 out of 70 staphylococcal isolates were positive by TCP, out of these 21.4% staphylococci were high biofilm producers, 62.8% staphylococci were moderate biofilm producers and 15.8% were non-biofilm producers. Maximum resistance was observed in biofilm producers to cotrimoxazole (74.5% and erythromycin (62.7% and none were resistant to vancomycin and linezolid. Out of total 59 biofilm producers, 20.3 % (12 were methicillin resistant and all these were S. aureus isolates. 19% (1 out of total 11 biofilm non-producers were methicillin resistant. Conclusion: Biofilm production was seen to be a major virulence factor in most of the staphylococcal isolates obtained from patients with signs and symptoms of septicaemia. S. aureus was found to be the major pathogen and timely detection of biofilm producing phenotype should be carried out using a simple and reproducible method, TCP which is both qualitative and quantitative.

Enterotoxin Producing Ability And Antimicrobial Susceptibility Of Coagulase-Negative Staphylococci Isolated From Goat Milk Cheese And Salted Yoghurt In Turkey

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S. Pehlivanlar Onen

2017-10-01

Full Text Available The aim of this study was to determine enterotoxin producing ability and antimicrobial susceptibility of coagulase negative staphylococci CNS in goats bulk milk cheese and salted yoghurt. CNS strains were identified by using GP card in VITEK 2 system. The presence of enterotoxins was determined by enzyme immunoassay test by using RIDASCREEN test kit. Antibiotic susceptibility in CNS strains was detected by using AST-P640 card in VITEK 2 system. A total of 100 CNS strains were isolated in 22 55 bulk milk samples and in 23 57.5 cheese samples. Staphylococcus spp. could not be isolated from salted yoghurt samples. The most encountered species were S. caprae 51.9 S. chromogenes 11.5 and S. xylosus 9.6 from milk samples and S. saprophyticus 60.4 S. xylosus 12.5 and S. haemolyticus 8.3 from cheese samples. Four CNS strains 4 isolated from samples were capable of producing enterotoxin. While all isolates were resistant to at least one antibiotic 74 of CNS strains showed resistance to two or more antibiotics. Enterotoxin production ability and high antibiotic resistance of the CNS strains isolated from goat bulk milk and cheese can lead to a risk for public health.

Prevalence of antiseptic-resistance genes in staphylococci isolated from orthokeratology lens and spectacle wearers in Hong Kong.

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Shi, Guang-sen; Boost, Maureen; Cho, Pauline

2015-05-01

To compare isolation of staphylococci from periorbital tissues and accessories of orthokeratology (ortho-k) lens and spectacle wearers and investigate prevalence of antiseptic-resistance (QAC) genes. To determine minimum inhibitory concentrations (MIC) of antiseptics and antibiotic susceptibility of isolates. Staphylococci were isolated from eyelids, eyelashes, and conjunctival sacs of 23 ortho-k lens wearers and 20 spectacle wearers. Samples were also collected from ortho-k lenses, lens cases, and spectacle frames. Isolations of Staphylococcus aureus were compared between ortho-k subjects and controls for all samples and for coagulase-negative staphylococci (CNS) from conjunctival sacs. QAC genes were amplified in 110 S. aureus and 59 CNS isolates and prevalence compared in isolates from ortho-k lens and spectacle wearers. Associations were assessed between presence of QAC genes and antibiotic and antiseptic susceptibility. Although isolation of S. aureus did not differ significantly in periorbital samples from ortho-k and control subjects, QAC genes were significantly more common in both S. aureus and CNS from ortho-k subjects (odds ratio 4.4 and 10.74, respectively). Overall, qacA/B was the predominant gene detected, being present in 26.5% CNS and 11% S. aureus. smr and qacH were present in 12% of CNS, but were less common in S. aureus. QAC gene-positive isolates had higher MICs to benzalkonium chloride and chlorhexidine digluconate. Our results suggest that long-term use of multipurpose solutions containing quaternary ammonium compounds may select for carriage of organisms harboring QAC genes. As these genes are associated with antibiotic resistance, their increased prevalence in isolates from contact lens wearers is a concern.

Comparative antibiogram of coagulase-negative Staphylococci (CNS) associated with subclinical and clinical mastitis in dairy cows

OpenAIRE

B. K. Bansal; D. K. Gupta; T. A. Shafi; S. Sharma

2015-01-01

Aim: The present study was planned to determine the in vitro antibiotic susceptibility of coagulase-negative Staphylococci (CNS) strains isolated from clinical and subclinical cases of mastitis in dairy cows. Antibiotic sensitivity profile will be helpful to recommend early therapy at the field level prior to availability of CST results. Materials and Methods: The milk samples from cases of clinical mastitis received in Mastitis Laboratory, Guru Angad Dev Veterinary and Animal Sciences Univer...

Modified DNA extraction for rapid PCR detection of methicillin-resistant staphylococci

International Nuclear Information System (INIS)

Japoni, A.; Alborzi, A.; Rasouli, M.; Pourabbas, B.

2004-01-01

Nosocomial infection caused by methicillin-resistant staphylococci poses a serious problem in many countries. The aim of this study was to rapidly and reliably detect methicillin-resistant-staphylococci in order to suggest appropriate therapy. The presence or absence of the methicillin-resistance gene in 115 clinical isolates of staphylococcus aureus and 50 isolates of coagulase negative staphylococci was examined by normal PCR. DNA extraction for PCR performance was then modified by omission of achromopeptadiase and proteinase K digestion, phenol/chloroform extraction and ethanol precipitation. All isolates with Mic>8 μ g/ml showed positive PCR. No differences in PCR detection have been observed when normal and modified DNA extractions have been performed. Our modified DNA extraction can quickly detect methicillin-resistant staphylococci by PCR. The advantage of rapid DNA extraction extends to both reduction of time and cost of PCR performance. This modified DNA extraction is suitable for different PCR detection, when staphylococci are the subject of DNA analysis

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

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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.

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.

Methicillin resistant staphylococci associated with bovine mastitis and their zoonotic importance

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S. Vishnupriya

2014-06-01

Full Text Available Aim: The present study was conducted to determine the zoonotic importance of methicillin resistant staphylococci associated with bovine mastitis and their potential role in transmission to animal handlers. Materials and Methods: A total of 158 milk samples from bovine mastitis cases and 126 nasal swabs from the animal handlers were sampled in and around Pondicherry (Southern India. The Presence of Staphylococcal organism was confirmed by PCR amplification using the genus specific primers and among the isolated Staphylococci; methicillin resistance was identified by genetic amplification of mec A methicillin resistant gene. Then the amplified gene from the bacteria expressing the mecA gene (PBP2a (~2kb fragment was further sequenced using four sets of primer pairs and aligned for determining their genetic relatedness between the sequences. Both phenotypic and genotypic analysis was carried out for the six MRS isolates (three bovine and three human in this study. Results: Out of 158 mastitis milk samples; 96 and 19 bovine isolates were found to be positive for Staphylococcal genus specific PCR and methicillin resistant (mecA gene PCR, respectively. Similarly, Out of 126 human nasal swabs, 64 and 13 human isolates were found to be positive for Staphylococcal genus specific PCR and mec A gene PCR, respectively. Among the 160 staphylococcal isolates (Bovine and Human origin; 51 were identified as coagulase-positive staphylococci (CPS and remaining as coagulase-negative staphylococci (CONS. The results obtained in this study revealed the presence of many species of Staphylococci but the predominant species were Staphylococcus aureus and S. epidermidis. The Sequence analysis of the mec A gene of human isolates obtained in this study had a maximum identity (99% -100% with the bovine isolates. Conclusion: The phenotypic and genotypic analysis carried out for the six MRS (Methicillin Resistant Staphylococci isolates in this study were indistinguishable

Antibiotic resistance and biofilm formation among coagulase-negative staphylococci isolated from clinical samples at a tertiary care hospital of eastern Nepal.

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Shrestha, Lok Bahadur; Bhattarai, Narayan Raj; Khanal, Basudha

2017-01-01

Coagulase negative staphylococci were long regarded non-pathogenic as they are the commensals of human skin and mucosa but the recent changes in the medical practice and changes in underlying host populations, they are being considered significant pathogens associated with number of nosocomial infections. The objective of the study was to determine the species, antimicrobial susceptibility pattern, biofilm forming ability of the clinically significant CoNS isolates and to compare the different methods for the detection of biofilm formation. A total of 52 clinically significant CoNS isolates obtained from different units during a year period were studied. Characterization was done using standard microbiological guidelines and antimicrobial susceptibility was done following CLSI guidelines. Biofilm formation was detected by using three methods i.e. tissue culture plate method, congo red agar method and tube adherence method. Among 52 isolates , S. epidermidis (52%) was the most common species which was followed by S. saprophyticus (18%) and S. haemolyticus (14%). Antimicrobial susceptibility pattern of CoNS documented resistance of 80% to ampicillin. Resistance to cefoxitin and ceftriaxone was observed in 58% of the isolates. Biofilm formation was observed in 65.38% of the isolates. The accuracy of Congo red agar and tube adherence method for the detection of biofilm formation was 82% and 76% respectively. CoNS isolates obtained from clinical samples should be processed routinely and antimicrobial susceptibility testing should be performed. Multidrug-resistant CoNS are prevalent. All the three methods i.e. tissue culture plate, Congo red agar and tube adherence method can be used in detecting biofilm formation.

Coagulase-negative staphylococci isolated from ocular wound infections after laser refractive surgery: attachment to and accumulation on soft contact lenses.

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Faghri, Jamshid; Razavi, Mohamad Reza

2009-03-01

Disposable soft contact lenses that are commonly used after laser refractive surgery are known to be colonized by bacteria and play a key role in Bacterial Keraitis (BK) pathogenesis. Coagulase-negative staphylococci (CoNS) have been found to be the most common pathogen involved in this postoperative infection. In this study a rapid and a simple assay was developed for studying attachment and accumulation of CoNS on soft contact lenses in vitro using [3H] thymidine. Thirty-five isolates of CoNS were obtained from 27 laser refractive surgery patients. Twenty-five of these thirty-five CoNS were isolated in multiple cultures. Ten CoNS were isolated in cultures from patients who underwent reoperation. The assay was optimized using a biofilm-producing strain, S. epidermidis RP62A, which was subcultured overnight at 37 degrees C on blood agar medium. Quantitative determination of biofilm production was tested. Presence of the genes icaADB and icaD was determined in all isolates. All isolates were biochemically analyzed using the Phene Plate (PhP) system modified for typing of CoNS. The CoNS isolates were further characterized to species level using ID32Staph.Mann-Whitney rank sum test and chi-square test were used to identify statistical differences in adherence, index, antibiotic susceptibility patterns, and biofilm production or presence of the ica operon between clinically significant isolates and non-postoperative BK isolates. No differences in attachment and accumulation were found between isolates causing BK after laser refractive surgery and contaminant isolates. In addition, there were no differences in the distribution of the ica operon between the two groups, as determined by polymerase chain reaction. Nevertheless, the ability to produce biofilm was found to be present significantly more frequently among BK isolates than among non-postoperative BK isolates. This study shows that the method using radioactive thymidine to analyze adherence of CoNS to soft

Coagulase-negative Staphylococci in Danish blood cultures: species distribution and antibiotic susceptibility.

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Jarløv, J O; Højbjerg, T; Busch-Sørensen, C; Scheibel, J; Møller, J K; Kolmos, H J; Wandall, D A

1996-03-01

The distribution and antibiotic susceptibility of coagulase-negative staphylococci (CoNS) isolated from blood cultures was examined in samples from hospitals covering most of Denmark. A total of 499 CoNS isolates were detected in 477 blood cultures from 340 patients and speciated as Staphylococcus epidermidis, 285; Staphylococcus hominis, 61; Staphylococcus haemolyticus, 43; Staphylococcus warneri, 12; Staphylococcus cohnii, 7; Staphylococcus saprophyticus, 4; Staphylococcus capitis, 2 and Staphylococcus lugdunensis, 1. Seventy-eight isolates could not be identified to species level and six were Micrococcus spp. In 108 (22.6%) blood culture sets, more than one CoNS strain were found, as detected by species identification, antibiogram and biotyping. Significantly more blood cultures from patients in university hospitals were drawn from central venous catheters. Comparing university and non-university hospitals, the overall antibiotic susceptibility among CoNS was only slightly different, except for methicillin and amikacin. The prevalence of methicillin-resistant strains was 35.1% in the university hospital strains vs. 25.3% in the non-university hospital strains. The overall prevalence of methicillin resistance was 32%. Great geographic variation in both species distribution and antibiotic resistance was observed. The high prevalence of S. epidermidis makes subtyping of this species important.

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

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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 log 10 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).

Genotypic and phenotypic β-lactam resistance and presence of PVL gene in Staphylococci from dry bovine udder.

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Vinodkumar Kulangara

Full Text Available Dairy cows affected with subclinical mastitis can be sources of virulent, antimicrobial-resistant Staphylococci to humans because of the excretion of the bacteria through their milk. This study focussed on the phenotypic and genotypic antibiotic resistance patterns of Staphylococci isolated from dairy cows in early dry period. Among 96 isolates of Gram positive cocci from 157 cows, 76 were identified as Coagulase Negative Staphylococci and the remaining 20 were Staphylococcus aureus. Typical amplicons of coagulase gene were obtained for all 20 samples of S. aureus with three major coagulase types being identified as giving 627 bp (40%, 910 bp (35% and 710 bp (25% long PCR products. The groEL gene was amplified in PCR of all 76 isolates of Coagulase Negative Staphylococci, and incubation of PCR products with restriction enzyme PvuII yielded three distinct PCR-RFLP fragment patterns bearing resemblance to S. chromogenes and S. hyicus. Highest sensitivity of Coagulase Negative Staphylococci was noted for Azithromycin (92.5% and the least to Tetracyclines (76.3%, whereas for S. aureus, it was Cefoperazone (95% and Azithromycin (72.2% respectively. Phenotypic resistance to Oxacillin (25 isolates, and Cefoxitin (11 isolates was detected by dilution method with a commercial strip (Ezy MICTM. Genotypic resistance to β-Lactam antibiotics was found in 65 (34 with mecA gene and 31 with blaZ gene isolates. Eighteen isolates possessed both the genes, with the PVL gene for virulence being detected in five of them. Nine isolates which had mecA gene were phenotypically susceptible to oxacillin while phenotypic resistance to oxacillin was observed in seven isolates that did not have either mecA or blaZ gene. This is the first report of persistent Staphylococcal infections possessing PVL gene and high level of genotypic resistance to β-Lactam antibiotics in small- holder dairy cattle from India.

Genotypic and phenotypic β-lactam resistance and presence of PVL gene in Staphylococci from dry bovine udder

Science.gov (United States)

Sivasailam, Asok; Sasidharan, Suchithra; Kollannur, Justin Davis; Syam, Radhika

2017-01-01

Dairy cows affected with subclinical mastitis can be sources of virulent, antimicrobial-resistant Staphylococci to humans because of the excretion of the bacteria through their milk. This study focussed on the phenotypic and genotypic antibiotic resistance patterns of Staphylococci isolated from dairy cows in early dry period. Among 96 isolates of Gram positive cocci from 157 cows, 76 were identified as Coagulase Negative Staphylococci and the remaining 20 were Staphylococcus aureus. Typical amplicons of coagulase gene were obtained for all 20 samples of S. aureus with three major coagulase types being identified as giving 627 bp (40%), 910 bp (35%) and 710 bp (25%) long PCR products. The groEL gene was amplified in PCR of all 76 isolates of Coagulase Negative Staphylococci, and incubation of PCR products with restriction enzyme PvuII yielded three distinct PCR-RFLP fragment patterns bearing resemblance to S. chromogenes and S. hyicus. Highest sensitivity of Coagulase Negative Staphylococci was noted for Azithromycin (92.5%) and the least to Tetracyclines (76.3%), whereas for S. aureus, it was Cefoperazone (95%) and Azithromycin (72.2%) respectively. Phenotypic resistance to Oxacillin (25 isolates), and Cefoxitin (11 isolates) was detected by dilution method with a commercial strip (Ezy MICTM). Genotypic resistance to β-Lactam antibiotics was found in 65 (34 with mecA gene and 31 with blaZ gene) isolates. Eighteen isolates possessed both the genes, with the PVL gene for virulence being detected in five of them. Nine isolates which had mecA gene were phenotypically susceptible to oxacillin while phenotypic resistance to oxacillin was observed in seven isolates that did not have either mecA or blaZ gene. This is the first report of persistent Staphylococcal infections possessing PVL gene and high level of genotypic resistance to β-Lactam antibiotics in small- holder dairy cattle from India. PMID:29091956

Antibiotic resistance and biofilm formation among coagulase-negative staphylococci isolated from clinical samples at a tertiary care hospital of eastern Nepal

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Lok Bahadur Shrestha

2017-08-01

Full Text Available Abstract Background Coagulase negative staphylococci were long regarded non-pathogenic as they are the commensals of human skin and mucosa but the recent changes in the medical practice and changes in underlying host populations, they are being considered significant pathogens associated with number of nosocomial infections. The objective of the study was to determine the species, antimicrobial susceptibility pattern, biofilm forming ability of the clinically significant CoNS isolates and to compare the different methods for the detection of biofilm formation. Methods A total of 52 clinically significant CoNS isolates obtained from different units during a year period were studied. Characterization was done using standard microbiological guidelines and antimicrobial susceptibility was done following CLSI guidelines. Biofilm formation was detected by using three methods i.e. tissue culture plate method, congo red agar method and tube adherence method. Results Among 52 isolates, S. epidermidis (52% was the most common species which was followed by S. saprophyticus (18% and S. haemolyticus (14%. Antimicrobial susceptibility pattern of CoNS documented resistance of 80% to ampicillin. Resistance to cefoxitin and ceftriaxone was observed in 58% of the isolates. Biofilm formation was observed in 65.38% of the isolates. The accuracy of Congo red agar and tube adherence method for the detection of biofilm formation was 82% and 76% respectively. Conclusion CoNS isolates obtained from clinical samples should be processed routinely and antimicrobial susceptibility testing should be performed. Multidrug-resistant CoNS are prevalent. All the three methods i.e. tissue culture plate, Congo red agar and tube adherence method can be used in detecting biofilm formation.

Public transport as a reservoir of methicillin-resistant staphylococci.

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Stepanović, S; Cirković, I; Djukić, S; Vuković, D; Svabić-Vlahović, M

2008-10-01

The aim of this study was to explore the occurrence of methicillin-resistant staphylococci in a large urban public transport system. Samples were taken from hand rails, which passengers hold onto when they are standing. In total, 1400 swabs taken from 55 vehicles (trolleybuses, trams and buses) were examined. As many as 30.1% samples were positive for the presence of methicillin-resistant coagulase-negative staphylococci (MRCoNS), but none for methicillin-resistant Staphylococcus aureus (MRSA). MRCoNS were isolated from all 55 vehicles. Nearly 50% of MRCoNS isolates displayed resistance not only to beta-lactams, but at least to two or more other classes of antimicrobials as well. This study demonstrated widespread occurrence of MRCoNS on hand rails in public transport vehicles. MRSA was not detected. The recovery of methicillin-resistant staphylococci from public transport system implies a potential risk for transmission of these bacteria in an out-hospital environment.

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

NARCIS (Netherlands)

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,

Prevalence and antimicrobial resistance of coagulase negative staphylococci clinical isolates from Ethiopia: a meta-analysis.

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Deyno, Serawit; Fekadu, Sintayehu; Seyfe, Sisay

2018-05-25

Antimicrobial resistant Coagulase-negative Staphylococci (CoNS) have limited treatment options, rendered diseases untreatable and made hospitals to be reservoirs of the resistant strains. The aim of this study was to estimate the pooled prevalence and antimicrobial resistance of clinical isolates of CoNS from Ethiopia. The electronic database search yielded 6511 articles of which 21 met predefined inclusion criteria. The pooled prevalence of CoNS from Ethiopia was 12% (95% confidence interval (CI): 8, 16%). The analyses revealed high level of CoNS resistance to methicilin (37%[95% CI: 21, 55%]), vancomycin (911%[95% CI: 0, 35%]), penicillin (58%[95% CI: 42, 74%]), amoxicillin (42%[95% CI: 23, 61%]), amoxicillin-clavulanate (27%[95% CI: 2, 27%]), ampicillin (64%[95% CI: 46, 80%]), tetracycline (60% [95% CI: 49, 70%]), doxycycline (36%[95% CI:19,55%]), Sulfamethoxazole-trimethoprim (50%[95% CI: 36, 64%]), ceftriaxone (27% [95% CI: 18, 38%]), cephalothin (32% [95% CI: 7, 62%]), norfloxacin (39%[95% CI: 24, 56%]), chloramphenicol (40%[95% CI: 23, 58%]), clindamycin (11% [95% CI: 2, 27%]), ciprofloxacin (14%[95% CI: 6, 22%]), gentamicin (27%[95% CI:19,36%]) and erythromycin (30%[95% CI:20%,42%]). High heterogeneity, I 2 ranging from 69.04 to 96.88%; p-values ≤0.01, was observed. Eggers' test did not detect publication bias for the meta-analyses and low risk of bias was observed in included studies. CoNS has gotten resistant to commonly used antimicrobials from Ethiopia. There is a need of launching national antimicrobial treatment, development and implementation of policy guidelines to contain the threat. Further research focusing on factors promoting resistance and the effect of resistance on treatment outcome studies are warranted.

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.

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.

Comparative antibiogram of coagulase-negative Staphylococci (CNS) associated with subclinical and clinical mastitis in dairy cows.

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Bansal, B K; Gupta, D K; Shafi, T A; Sharma, S

2015-03-01

The present study was planned to determine the in vitro antibiotic susceptibility of coagulase-negative Staphylococci (CNS) strains isolated from clinical and subclinical cases of mastitis in dairy cows. Antibiotic sensitivity profile will be helpful to recommend early therapy at the field level prior to availability of CST results. The milk samples from cases of clinical mastitis received in Mastitis Laboratory, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana and those of subclinical mastitis collected during routine screening of state dairy farms, were subjected to microbial culture. Identification of CNS organisms was done by standard biochemical tests. Antibiotic sensitivity testing, based on 30 antibiotics belonging to 12 groups, was done on 58 randomly selected CNS isolates (clinical isolates: 41, subclinical isolates: 17). Isolates were highly susceptible to chloramphenicol (98.3%), gentamicin (93.1%), streptomycin (91.4%), linezolid (91.4%), ceftixozime (87.9%), cloxacillin (86.2%), clotrimazole (86.2%), bacitracin (86.2%), enrofloxacin (84.5%) and ceftrioxone + tazobactum (70.7%), while resistance was observed against amoxicillin (77.6%), penicillin (75.9%), ampicillin (74.1%) and cefoperazone (51.7%). Overall, isolates from clinical cases of mastitis had a higher resistance than subclinical isolates. CNS isolates were susceptible to chloramphenicol, gentamicin and streptomycin, while higher resistance was recorded against routinely used penicillin group.

Antimicrobial resistance and detection of the mecA gene besides enterotoxin-encoding genes among coagulase-negative Staphylococci isolated from clam meat of Anomalocardia brasiliana.

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Batista, Jacqueline Ellen Camelo; Ferreira, Ewerton Lucena; Nascimento, Danielle Cristina de Oliveira; Ventura, Roberta Ferreira; de Oliveira, Wagner Luis Mendes; Leal, Nilma Cintra; Lima-Filho, José Vitor

2013-12-01

The marine clam Anomalocardia brasiliana is a candidate as a sentinel animal to monitor the contamination levels of coliforms in shellfish-harvesting areas of Brazil's northeastern region. The aim of the present study was to search enterotoxin-encoding genes plus the mecA gene among coagulase-negative staphylococci (CNS) isolates from shellfish meats of A. brasiliana. The specimen clam (n=48; 40 clams per sample) was collected during low tide in the bay area of Mangue Seco from April through June 2009, and random samples of chilled and frozen shelled clam meat (n=33; 250 g per sample) were obtained from retail shops from January through March 2012. Seventy-nine CNS isolates were identified, including Staphylococcus xylosus, S. cohnii spp. urealyticus, S. sciuri, and S. lentus. A high percentage of isolates resistant to erythromycin (58.5%), penicillin (51.2%), and tetracycline (43.9%), and the fluoroquinolones levofloxacin (39%) and ciprofloxacin (34.1%) were recorded from those environmental samples. Isolates from retail shops were particularly resistant to oxacillin (55.3%) and penicillin (36.8%). All CNS resistant to oxacillin and/or cefoxitin were positive for the presence of the mecA gene, but phenotypically susceptible to vancomycin. Also, the enterotoxin-encoding genes seg and seh were detected through multiplex-polymerase chain reaction in 77.7% and 88.8% of the isolates from environmental samples, versus 90.5% and 100% of the isolates from retail shops, respectively. The data reveal the risk to public health due to consuming raw or undercooked shellfish containing enterotoxigenic plus methicillin-resistant CNS.

Antimicrobial susceptibility profile and research of mec A and erm genes in coagulase-negative staphylococci isolated from platelet concentrates bags

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Rosiéli Martini

2017-04-01

Full Text Available Abstract In recent years, several studies have described the clinical impact of bacterial infection associated with transfusion of platelet concentrates (PCs. Among the blood components, PCs are responsible for the highest rates of bacterial contamination as well as septic transfusion reactions. We assessed antimicrobial susceptibility profile, resistance to methicillin (MRCoNS, and resistance to macrolides, lincosamides and streptogramins of group B (MLSB of 16 coagulase-negative staphylococci (CoNS isolates from an investigation in 691 PCs bags. We then compared conventional and automated phenotypic methods, disc diffusion test (DD and VITEK(r 2, respectively as well as phenotypic and genotypic methods (Polymerase Chain Reaction - PCR. All CoNS were susceptible to vancomycin. The disc diffusion test characterized 18.75% as MRCoNS and 37.5% with inducible resistance to MLSB (iMLSB, and with VITEK(r 2, 6.3% and 31.25%, respectively. The mecA gene was detected in 18.75% and the erm gene in 31.25% of the isolates. In this study, we found equal percentage values between presence of the mecA gene by PCR and resistance to methicillin using cefoxitin by DD test, evidence of the erm gene by PCR, and iMLSB resistance by automation (VITEK(r 2. Moreover, we identified three strains with beta-lactamase overproduction, and the occurrence of a bigger mistake was verified when automation was compared with DD test. And we observed that D-test was the most reliable for the detection of iMLSB resistance in Staphylococcus sp.

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.

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.

Comparative antibiogram of coagulase-negative Staphylococci (CNS associated with subclinical and clinical mastitis in dairy cows

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B. K. Bansal

2015-03-01

Full Text Available Aim: The present study was planned to determine the in vitro antibiotic susceptibility of coagulase-negative Staphylococci (CNS strains isolated from clinical and subclinical cases of mastitis in dairy cows. Antibiotic sensitivity profile will be helpful to recommend early therapy at the field level prior to availability of CST results. Materials and Methods: The milk samples from cases of clinical mastitis received in Mastitis Laboratory, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana and those of subclinical mastitis collected during routine screening of state dairy farms, were subjected to microbial culture. Identification of CNS organisms was done by standard biochemical tests. Antibiotic sensitivity testing, based on 30 antibiotics belonging to 12 groups, was done on 58 randomly selected CNS isolates (clinical isolates: 41, subclinical isolates: 17. Results: Isolates were highly susceptible to chloramphenicol (98.3%, gentamicin (93.1%, streptomycin (91.4%, linezolid (91.4%, ceftixozime (87.9%, cloxacillin (86.2%, clotrimazole (86.2%, bacitracin (86.2%, enrofloxacin (84.5% and ceftrioxone + tazobactum (70.7%, while resistance was observed against amoxicillin (77.6%, penicillin (75.9%, ampicillin (74.1% and cefoperazone (51.7%. Overall, isolates from clinical cases of mastitis had a higher resistance than subclinical isolates. Conclusion: CNS isolates were susceptible to chloramphenicol, gentamicin and streptomycin, while higher resistance was recorded against routinely used penicillin group.

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

Comparative study using phenotypic, genotypic and proteomics methods for identification of coagulase-negative staphylococci

NARCIS (Netherlands)

Dr. P.F.G. Wolffs; Ing M. Valkenburg; Dr. A.J.C. van den Brule, van den; M.Sc. A. Jansz; Drs A.J.M. Loonen; Ing J.N.B. Bergland

2012-01-01

Five methods were compared to determine the best technique for accurate identification of coagulase-negative staphylococci (CoNS) (n=142 strains). MALDI-TOF MS showed the best results for rapid and accurate CoNS differentiation (correct identity in 99.3%). An alternative to this approach could be

Coagulase-Negative Staphylococci in Human Milk From Mothers of Preterm Compared With Term Neonates.

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Soeorg, Hiie; Metsvaht, Tuuli; Eelmäe, Imbi; Metsvaht, Hanna Kadri; Treumuth, Sirli; Merila, Mirjam; Ilmoja, Mari-Liis; Lutsar, Irja

2017-05-01

Human milk is the preferred nutrition for neonates and a source of bacteria. Research aim: The authors aimed to characterize the molecular epidemiology and genetic content of staphylococci in the human milk of mothers of preterm and term neonates. Staphylococci were isolated once per week in the 1st month postpartum from the human milk of mothers of 20 healthy term and 49 preterm neonates hospitalized in the neonatal intensive care unit. Multilocus variable-number tandem-repeats analysis and multilocus sequence typing were used. The presence of the mecA gene, icaA gene of the ica-operon, IS 256, and ACME genetic elements was determined by PCR. The human milk of mothers of preterm compared with term neonates had higher counts of staphylococci but lower species diversity. The human milk of mothers of preterm compared with term neonates more often contained Staphylococcus epidermidis mecA (32.7% vs. 2.6%), icaA (18.8% vs. 6%), IS 256 (7.9% vs. 0.9%), and ACME (15.4% vs. 5.1%), as well as Staphylococcus haemolyticus mecA (90.5% vs. 10%) and IS 256 (61.9% vs. 10%). The overall distribution of multilocus variable-number tandem-repeats analysis (MLVA) types and sequence types was similar between the human milk of mothers of preterm and term neonates, but a few mecA-IS 256-positive MLVA types colonized only mothers of preterm neonates. Maternal hospitalization within 1 month postpartum and the use of an arterial catheter or antibacterial treatment in the neonate increased the odds of harboring mecA-positive staphylococci in human milk. Limiting exposure of mothers of preterm neonates to the hospital could prevent human milk colonization with more pathogenic staphylococci.

Bacteriological Study of 100 Cases of Pyodermas with Special Reference to Staphylococci, Their Antibiotic Sensitivity and Phage Pattern

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T V Ramani

1980-01-01

Full Text Available One hundred new cases of pyodenna attending King George Hospital, Vishakhapatnam were investigated bacteriologically with antibiotic sensitivity of all the strains isolated and phage typing of coagulm positive staphylocci. Among these 50 had impetigo and 15 each ahd furunculosis and fouiculitis. The remaining included various other clinical entities. Children under 10 years were observed to have high incidence of pyoderma. A total of 88 strains of staphylococci (77 coagulase positive and 11 coagulase negative strains 25 strains of beta haemolytic streptococci and 3 strains of Klebsiella were isolated. Staphylococci were found to be the commonest aetiological agents either single or in association with other organisms. Of the 76 strains of coagulase positive staphylococci 32 strains were not phage typable and among the 44 typable strains 17 (38% belonged to group III and 15 (36.5% to mixed group. Coagulase positive staphylococci showed high sensitivity to garamycin, kanamycin and erythromycin and high resistance to penicillin and streptomycin. Multiple drug resistance was also high among these strains. Coagulase negative staphylococci were found to be more sensitive with less incidence of multiple drug resistance. Most of multiple drug resistant strains belonged to group III phage types. Beta haemolytic streptococci were found to be highly sensitive to all the antibiotics tested.

Methicillin resistance gene diversity in staphylococci isolated from captive and free-ranging wallabies

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Michelle M. S. Chen

2016-05-01

Full Text Available Background: Infection with methicillin-resistant staphylococci (MRS can be life-threatening in humans and its presence in animals is a cause for public health concern. The aim of this study was to measure the prevalence of MRS in captive and free-ranging wallabies over a 16-month period in South Australia, Australia. Materials and methods: Eighty-nine purified staphylococcal isolates recovered from 98 captive and free-ranging wallabies' anterior nasal swabs were used in this study. All isolates were tested for the presence of the mecA, mecA1, and mecC genes. Multiplex PCR-directed SCCmec-typing, ccrB-typing, and determination of the minimal inhibitory concentration of oxacillin were performed on mec-positive isolates. Results and discussion: In total, 11 non-Staphylococcus aureus MRS were isolated from 7 out of 98 animals, corresponding to a 7.1% carriage rate. The SCCmec types I, III, and V were identified by multiplex PCR and sequencing of the ccrB gene. This is the first report of MRS carriage in both captive and free-ranging wallabies in Australia. These data demonstrate a low prevalence of MRS and no association between wallaby captivity status and MRS carriage could be assigned. These animals may act as a reservoir for the exchange of genetic elements between staphylococci. Furthermore, the mecA genes of animal isolates were identical to that found in human MRS strains and thus the possibility of zoonotic transfer must be considered.

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

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

2008-01-01

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. PMID:24031279

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

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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. © 2016 Institute of Food Technologists®

Diversity and antimicrobial susceptibility profiling of staphylococci isolated from bovine mastitis cases and close human contacts.

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Schmidt, T; Kock, M M; Ehlers, M M

2015-09-01

The objectives of this study were to examine the diversity of Staphylococcus spp. recovered from bovine intramammary infections and humans working in close contact with the animals and to evaluate the susceptibility of the staphylococcal isolates to different antimicrobials. A total of 3,387 milk samples and 79 human nasal swabs were collected from 13 sampling sites in the KwaZulu-Natal province of South Africa. In total, 146 Staph. aureus isolates and 102 coagulase-negative staphylococci (CNS) were recovered from clinical and subclinical milk samples. Staphylococcusaureus was isolated from 12 (15.2%) of the human nasal swabs and 95 representative CNS were recovered for further characterization. The CNS were identified using multiplex-PCR assays, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and tuf gene sequencing. Seven Staphylococcus spp. were identified among the CNS of bovine origin, with Staph.chromogenes (78.4%) predominating. The predominant CNS species recovered from the human nasal swabs was Staph.epidermidis (80%) followed by Staph.chromogenes (6.3%). The antimicrobial susceptibility of all staphylococcal isolates was evaluated using disk diffusion and was supplemented by screening for specific antimicrobial resistance genes. Ninety-eight (67.1%) Staph.aureus isolates of bovine origin were pansusceptible; 39 (26.7%) isolates were resistant to a single class, and 7 (4.8%) isolates were resistant to 2 classes of antimicrobials. Two Staph. aureus (1.4%) isolates were multidrug-resistant. Resistance to penicillin was common, with 28.8% of the bovine and 75% of the human Staph. aureus isolates exhibiting resistance. A similar observation was made with the CNS, where 37.3% of the bovine and 89.5% of the human isolates were resistant to penicillin. Multidrug-resistance was common among the human CNS, with 39% of the isolates exhibiting resistance to 3 or more classes of antimicrobials. The antimicrobial

Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices.

OpenAIRE

Christensen, G D; Simpson, W A; Younger, J J; Baddour, L M; Barrett, F F; Melton, D M; Beachey, E H

1985-01-01

The adherence of coagulase-negative staphylococci to smooth surfaces was assayed by measuring the optical densities of stained bacterial films adherent to the floors of plastic tissue culture plates. The optical densities correlated with the weight of the adherent bacterial film (r = 0.906; P less than 0.01). The measurements also agreed with visual assessments of bacterial adherence to culture tubes, microtiter plates, and tissue culture plates. Selected clinical strains were passed through ...

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.

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

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Ugwu, Clifford C; Gomez-Sanz, Elena; Agbo, Ifeoma C; Torres, Carmen; Chah, Kennedy F

2015-01-01

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, cat pC221, and cat pC223. 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.

A Review of Staphylococcal Cassette Chromosome mec (SCCmec) Types in Coagulase-Negative Staphylococci (CoNS) Species.

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Saber, Huda; Jasni, Azmiza Syawani; Jamaluddin, Tengku Zetty Maztura Tengku; Ibrahim, Rosni

2017-10-01

Coagulase-negative staphylococci (CoNS) are considered low pathogenic organisms. However, they are progressively causing more serious infections with time because they have adapted well to various antibiotics owing to their ability to form biofilms. Few studies have been conducted on CoNS in both, hospital and community-acquired settings, especially in Malaysia. Thus, it is important to study their species and gene distributions. A mobile genetic element, staphylococcal cassette chromosome mec (SCC mec ), plays an important role in staphylococci pathogenesis. Among CoNS, SCC mec has been studied less frequently than Staphylococcus aureus (coagulase-positive staphylococci). A recent study (8) conducted in Malaysia successfully detected SCC mec type I to VIII as well as several new combination patterns in CoNS species, particularly Staphylococcus epidermidis . However, data are still limited, and further research is warranted. This paper provides a review on SCC mec types among CoNS species.

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

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...... 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...

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.

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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. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

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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

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.

Tetracycline resistance phenotypes and genotypes of coagulase-negative staphylococcal isolates from bubaline mastitis in Egypt.

Science.gov (United States)

El-Razik, K A Abd; Arafa, A A; Hedia, R H; Ibrahim, E S

2017-06-01

This study was devoted to elucidate the tetracycline resistance of coagulase-negative staphylococci (CNS) derived from normal and subclinical mastitic (SCM) buffaloes' milk in Egypt. A total of 81 milk samples from 46 normal buffalo milk samples and 35 SCM buffalo milk samples at private dairy farms of Egypt were used in this study. CNS were identified using phenotypic and molecular methods (polymerase chain reaction [PCR]). CNS isolates were tested for tetracycline resistance using routine methods and multiplex PCR targeting tetracycline ( tet ) resistance genes followed by sequencing of positive PCR products and phylogenetic analysis. Isolation and identification of 28 (34.5%) CNS from normal and SCM buffaloes' milk, namely, Staphylococcus intermedius (39.2%), Staphylococcus xylosus (25.0%), Staphylococcus epidermidis (10.7%), Staphylococcus hominis (10.7%), and 3.5% to each of Staphylococcus sciuri , Staphylococcus hyicus , Staphylococcus lugdunensis , and Staphylococcus simulans . Using nested PCR, all the 28 CNS isolates revealed positive for 16srRNA gene specific for genus staphylococci and negative for thermonuclease ( nuc ) gene specific for Staphylococcus aureus species. The presence of tetracycline resistance-encoding genes ( tet K, tet L, tet M, and tet O) was detected by multiplex PCR. All isolates were negative for tet L, M, and O genes while 14 (50%) CNS isolates were positive for tet K gene, namely, S. lugdunensis (100%), S. hominis (100%), S. epidermidis (66.6%), S. intermedius (45.4%), and S. xylosus (42.8%). Nucleotide sequencing of tet K gene followed by phylogenetic analysis showed the high homology between our CNS isolates genes of tetracycline resistance with S. aureus isolates including Egyptian ones. This proves the transfer of the tetracycline resistance encoding genes between coagulase-negative and coagulase positive Staphylococcus spp. CNS isolates have distinguishingly high resistance to tetracycline. Abundant tetracycline usage for

Tetracycline resistance phenotypes and genotypes of coagulase-negative staphylococcal isolates from bubaline mastitis in Egypt

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K. A. Abd El-Razik

2017-06-01

Full Text Available Aim: This study was devoted to elucidate the tetracycline resistance of coagulase-negative staphylococci (CNS derived from normal and subclinical mastitic (SCM buffaloes' milk in Egypt. Materials and Methods: A total of 81 milk samples from 46 normal buffalo milk samples and 35 SCM buffalo milk samples at private dairy farms of Egypt were used in this study. CNS were identified using phenotypic and molecular methods (polymerase chain reaction [PCR]. CNS isolates were tested for tetracycline resistance using routine methods and multiplex PCR targeting tetracycline (tet resistance genes followed by sequencing of positive PCR products and phylogenetic analysis. Results: Isolation and identification of 28 (34.5% CNS from normal and SCM buffaloes' milk, namely, Staphylococcus intermedius (39.2%, Staphylococcus xylosus (25.0%, Staphylococcus epidermidis (10.7%, Staphylococcus hominis (10.7%, and 3.5% to each of Staphylococcus sciuri, Staphylococcus hyicus, Staphylococcus lugdunensis, and Staphylococcus simulans. Using nested PCR, all the 28 CNS isolates revealed positive for 16srRNA gene specific for genus staphylococci and negative for thermonuclease (nuc gene specific for Staphylococcus aureus species. The presence of tetracycline resistance-encoding genes (tetK, tetL, tetM, and tetO was detected by multiplex PCR. All isolates were negative for tetL, M, and O genes while 14 (50% CNS isolates were positive for tetK gene, namely, S. lugdunensis (100%, S. hominis (100%, S. epidermidis (66.6%, S. intermedius (45.4%, and S. xylosus (42.8%. Nucleotide sequencing of tetK gene followed by phylogenetic analysis showed the high homology between our CNS isolates genes of tetracycline resistance with S. aureus isolates including Egyptian ones. This proves the transfer of the tetracycline resistance encoding genes between coagulase-negative and coagulase-positive Staphylococcus spp. Conclusion: CNS isolates have distinguishingly high resistance to tetracycline

Emergence of linezolid-resistant coagulase-negative staphylococci in an intensive care unit.

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Balandin, Bárbara; Lobo, Beatriz; Orden, Beatriz; Román, Federico; García, Elena; Martínez, Rocío; Valdivia, Miguel; Ortega, Alfonso; Fernández, Inmaculada; Galdos, Pedro

2016-01-01

The aim of this study was to report the emergence of linezolid-resistant coagulase-negative staphylococci (CoNS) in an intensive care unit. An observational study was conducted in critically ill patients with colonization or infection by linezolid-resistant CoNS between January 2010 and December 2014. We analyzed the epidemiological and clinical features, and the mechanism of resistance to linezolid. We also evaluated the association between the incidence of linezolid-resistant CoNS strains and the consumption of linezolid in the study period. During the study period 49 patients had a linezolid-resistant CoNS strain isolated from clinical samples (blood in 42 cases, urine in 6, peritoneal fluid in 1). Molecular study showed a combination of mechanisms of resistance. Most patients were critically ill (APACHE II score = 21.9 ± 8.3) and nearly all had undergone surgery and invasive procedures, and had prior exposure to antibiotics. Linezolid-resistant CoNS were considered to be contaminants in 42 patients and associated with infection in 7 patients, comprising bacteremia and septic shock in most of them. They were successfully treated with glycopeptides or daptomycin. A modest significant correlation was observed between the decrease in linezolid consumption and the lower incidence of resistant isolates. Linezolid-resistant CoNS had emerged in critically ill patients with severe underlying diseases and prior antibiotic exposure. Most isolates represented colonization; however, linezolid-resistant CoNS can produce serious infections in critically ill patients. Glycopeptides and daptomycin seem to provide useful alternatives for therapy of these infections. A relationship was found between linezolid consumption and the incidence of linezolid-resistant CoNS strains.

Interaction between bovine-associated coagulase-negative staphylococci species and strains and bovine mammary epithelial cells reflects differences in ecology and epidemiological behavior.

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Souza, F N; Piepers, S; Della Libera, A M M P; Heinemann, M B; Cerqueira, M M O P; De Vliegher, S

2016-04-01

Bacteria adherence seems to be an essential first stage for the internalization of bacteria into the cytoplasm of the host cell, which is considered an important virulence strategy enabling bacteria to occupy a microenvironment separated from host defense mechanisms. Thus, this study aimed to explore the difference in the capacity of 4 bovine-associated staphylococci species or strains to adhere to and internalize into bovine mammary epithelial cells (MEC). Three different isolates of coagulase-negative staphylococci (CNS) were used: one strain of Staphylococcus fleurettii isolated from sawdust and considered an environmental opportunistic bacterium, and 2 dissimilar Staphylococcus chromogenes isolates, one cultured from a heifer's teat apex (Staph. chromogenes TA) and the other originating from a chronic intramammary infection (Staph. chromogenes IM). Also, one well-characterized strain of Staphylococcus aureus (Newbould 305) was used for comparison with a major mastitis pathogen. The CNS species and strains adhered to and internalized into MEC slower than did Staph. aureus. Still, we observed high variation in adhesion and internalization capacity among the different CNS, with Staph. chromogenes IM showing a greater ability to adhere to and internalize into MEC than the 2 CNS strains isolated from extramammary habitats. In conclusion, the 3 well-characterized bovine-associated CNS species and strains originating from distinct habitats showed clear differences in their capacity to adhere to and internalize into MEC. The observed differences might be related to their diversity in ecology and epidemiological behavior. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Sepsis due to linezolid resistant Staphylococcus cohnii and Staphylococcus kloosii: first reports of linezolid resistance in coagulase negative staphylococci from India.

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Peer, M A; Nasir, R A; Kakru, D K; Fomda, B A; Bashir, G; Sheikh, I A

2011-01-01

Linezolid, a viable alternative to vancomycin against methicillin resistant staphylococcal isolates, has been in use for a decade around the globe. However, resistance against staphylococci remains extremely rare and unreported from most of the Asian countries. Herein, we report two cases of linezolid resistant, coagulase negative staphylococcal sepsis for the first time from India. The first case was an 18-year-old burn patient, who, after a major graft surgery, landed in sepsis, and linezolid resistant Staphylococcus cohnii with an minimum inhibitory concentration (MIC) of >256 μg/ml by both broth microdilution and Etest, was isolated from multiple blood cultures. The second patient was a 60-year-old male with an intracranial bleed and sepsis, from whose blood cultures, linezolid resistant Staphylococcus kloosii was repeatedly isolated. Linezolid MIC was >32 μg/ml by broth microdilution and >16 μg/ml by Etest.

Sepsis due to linezolid resistant Staphylococcus cohnii and Staphylococcus kloosii: First reports of linezolid resistance in coagulase negative staphylococci from India

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M A Peer

2011-01-01

Full Text Available Linezolid, a viable alternative to vancomycin against methicillin resistant staphylococcal isolates, has been in use for a decade around the globe. However, resistance against staphylococci remains extremely rare and unreported from most of the Asian countries. Herein, we report two cases of linezolid resistant, coagulase negative staphylococcal sepsis for the first time from India. The first case was an 18-year-old burn patient, who, after a major graft surgery, landed in sepsis, and linezolid resistant Staphylococcus cohnii with an minimum inhibitory concentration (MIC of >256 μg/ml by both broth microdilution and Etest, was isolated from multiple blood cultures. The second patient was a 60-year-old male with an intracranial bleed and sepsis, from whose blood cultures, linezolid resistant Staphylococcus kloosii was repeatedly isolated. Linezolid MIC was >32 μg/ml by broth microdilution and >16 μg/ml by Etest.

High-resolution melt analysis for species identification of coagulase-negative staphylococci derived from bovine milk.

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Ajitkumar, Praseeda; Barkema, Herman W; Zadoks, Ruth N; Morck, Douglas W; van der Meer, Frank J U M; De Buck, Jeroen

2013-03-01

Coagulase-negative staphylococci (CNS) are the most frequently isolated pathogens isolated from bovine milk. In this study, we report a rapid assay for species identification of CNS using high-resolution melt analysis (HRMA) of 16S rDNA sequences. Real-time polymerase chain reaction amplification of 16S rRNA gene fragment, spanning the variable region V1 and V2, was performed with a resulting amplicon of 215 bp. A library of distinct melt curves of reference strains of 13 common CNS species was created using HRMA. Sequencing of 16S rRNA and rpoB genes, and, when needed, tuf gene, of 100 CNS isolates obtained from Canadian Bovine Mastitis Research Network was done to determine their species identity, allowing for subsequent evaluation of the performance of HRMA for field isolates of bovine CNS. A combination of HRMA and sequencing revealed that Staphylococcus chromogenes, S. xylosus, S. simulans, and S. sciuri had multiple genotypes, complicating their resolution by HRMA. As the 3 genotypes of S. chromogenes had distinct melt curves, the 3 distinct genotypes were employed as reference strains in a blinded trial of 156 CNS isolates to identify S. chromogenes. HRMA correctly identified all S. chromogenes isolates which were later confirmed by sequencing. Staphylococcus chromogenes (68%) was most frequently found among the CNS isolates, followed by S. haemolyticus (10%) and S. xylosus (6%). The present study revealed that HRMA of 16S rRNA gene (V1-V2) could be used as a rapid, efficient, low-cost, and minimally cumbersome technique for S. chromogenes identification, the most common CNS derived from bovine milk. Copyright © 2013 Elsevier Inc. All rights reserved.

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

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.

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

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M. N. Boukhatem

2015-05-01

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.

Species Identification And Antibiotic Susceptibilities Of Coagulase- Negative Staphylococci Isolated From Urinary Tract Infection Specimens

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Hashmi, A.; Abdullah, F. E.

2016-01-01

Objective: To determine the incidence of Coagulase- negative S. aureusin urinary tract infections and sensitivities of these isolates to antimicrobial agents. Study Design: Cohort study. Place and Duration of Study: Dr. Essa Laboratory and Immunology and Infectious Disease Research Laboratory (IIDRL), Microbiology Department, University of Karachi, from January 2009 to January 2010. Methodology: Urine specimens, suggestive of urinary tract infection (UTI), were identified. Speciation of isolates was done using API-20 Staph.system. Screening of extracellular products was done using SDS-PAGE electrophoresis and Hemolysin on blood-agar plates. Minimum inhibitory concentration (MICs) of antibiotics was estimated by microtiter well plate method. Frequency and percentages were determined and chi-square test was used for comparing proportions with significance at p < 0.05. Results: Coagulase - negative S. aureus(CONS) were the cause of urinary tract infection in 56 out of 1866 outpatient (3 percent) and 164 of 1261 in patient (13 percent), urinary tract infections (p < 0.001). Two hundred and twenty CONS isolates were identified. The most common CONS identified was S. saprophyticus (31 percent, 68 strains). The relative frequency of Coagulase - negative S. aureus was 6 percent (13 strains). All isolates were sensitive to Vancomycin and Linezolid. Resistance was 69 percent to Ampicillin, 53 percent to Methicillin, and 37.5 percent to Ciprofloxacin. Conclusion: CONS are a potential uropathogens, with capability of slime production and resistance to common empirical prescriptions. This also warrants formulation of an appropriate antibiotic policy that covers CONS. (author)

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.

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Cirkovic, Ivana; Bozic, Dragana D; Draganic, Veselin; Lozo, Jelena; Beric, Tanja; Kojic, Milan; Arsic, Biljana; Garalejic, Eliana; Djukic, Slobodanka; Stankovic, Slavisa

2016-01-01

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. 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. 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 < 0.05, respectively), while BGBU1-4 crude extract inhibited biofilm formation by all L. monocytogenes isolates (p < 0.01 and p < 0.05, respectively). Both bacteriocins in concentrations of 100 AU/mL and 200 AU/mL reduced the amount of 24 h old CoNS and L. monocytogenes biofilms (p < 0.05, p < 0.01, p < 0.001). This study suggests that novel 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.

Genetic characterization of antimicrobial resistance in coagulase-negative staphylococci from bovine mastitis milk.

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Frey, Yvonne; Rodriguez, Joan Peña; Thomann, Andreas; Schwendener, Sybille; Perreten, Vincent

2013-04-01

Coagulase-negative staphylococci (CNS; n=417) were isolated from bovine milk and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Nineteen different species were identified, and Staphylococcus xylosus, Staphylococcus chromogenes, Staphylococcus haemolyticus, and Staphylococcus sciuri were the most prevalent species. Resistance to oxacillin (47.0% of the isolates), fusidic acid (33.8%), tiamulin (31.9%), penicillin (23.3%), tetracycline (15.8%), streptomycin (9.6%), erythromycin (7.0%), sulfonamides (5%), trimethoprim (4.3%), clindamycin (3.4%), kanamycin (2.4%), and gentamicin (2.4%) was detected. Resistance to oxacillin was attributed to the mecA gene in 9.7% of the oxacillin-resistant isolates. The remaining oxacillin-resistant CNS did not contain the mecC gene or mecA1 promoter mutations. The mecA gene was detected in Staphylococcus fleurettii, Staphylococcus epidermidis, Staph. haemolyticus, and Staph. xylosus. Resistance to tetracycline was attributed to the presence of tet(K) and tet(L), penicillin resistance to blaZ, streptomycin resistance to str and ant(6)-Ia, and erythromycin resistance to erm(C), erm(B), and msr. Resistance to tiamulin and fusidic acid could not be attributed to an acquired resistance gene. In total, 15.1% of the CNS isolates were multidrug resistant (i.e., resistant to 2 or more antimicrobials). The remaining CNS isolates were susceptible to antimicrobials commonly used in mastitis treatment. Methicillin-resistant CNS isolates were diverse, as determined by mecA gene sequence analysis, staphylococcal cassette chromosome mec typing, and pulsed-field gel electrophoresis. Arginine catabolic mobile element types 1 and 3 were detected in both methicillin-resistant and methicillin-susceptible Staph. epidermidis and were associated with sequence types ST59 and ST111. Because this study revealed the presence of multidrug-resistant CNS in a heterogeneous CNS population, we recommend antibiogram analysis

LAB Bacteriocins Controlling the Food Isolated (Drug-Resistant Staphylococci

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Jesús Perales-Adán

2018-06-01

Full Text Available Staphylococci are a group of microorganisms that can be often found in processed food and they might pose a risk for human health. In this study we have determined the content of staphylococci in 7 different fresh goat-milk cheeses. These bacteria were present in all of them, ranging from 103 to 106 CFU/g based on growth on selective media. Thus, a set of 97 colonies was randomly picked for phenotypic and genotypic identification. They could be clustered by RAPD-PCR in 10 genotypes, which were assigned by 16S rDNA sequencing to four Staphylococcus species: Staphylococcus aureus, Staphylococcus chromogenes, S. simulans, and S. xylosus. Representative strains of these species (n = 25 were tested for antibiotic sensitivity, and 11 of them were resistant to at least one of the antibiotics tested, including erythromycin, amoxicillin-clavulanic acid and oxacillin. We also tested two bacteriocins produced by lactic acid bacteria (LAB, namely the circular bacteriocin AS-48 and the lantibiotic nisin. These peptides have different mechanism of action at the membrane level. Nevertheless, both were able to inhibit staphylococci growth at low concentrations ranging between 0.16–0.73 μM for AS-48 and 0.02–0.23 μM for nisin, including the strains that displayed antibiotic resistance. The combined effect of these bacteriocins were tested and the fractional inhibitory concentration index (FICI was calculated. Remarkably, upon combination, they were active at the low micromolar range with a significant reduction of the minimal inhibitory concentration. Our data confirms synergistic effect, either total or partial, between AS-48 and nisin for the control of staphylococci and including antibiotic resistant strains. Collectively, these results indicate that the combined use of AS-48 and nisin could help controlling (pathogenic staphylococci in food processing and preventing antibiotic-resistant strains reaching the consumer in the final products.

Clinical and molecular features of methicillin-resistant, coagulase-negative staphylococci of pets and horses.

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Kern, Andrea; Perreten, Vincent

2013-06-01

To determine the antibiotic resistance and fingerprint profiles of methicillin-resistant coagulase-negative staphylococci (MRCoNS) from animal infections among different practices and examine the history of antibiotic treatment. Isolates were identified by mass spectrometry and tested for antimicrobial resistance by broth dilution, microarrays and sequence analysis of the topoisomerases. Diversity was assessed by PFGE, icaA PCR and staphylococcal cassette chromosome mec (SCCmec), arginine catabolic mobile element (ACME) and multilocus sequence typing. Clinical records were examined retrospectively. MRCoNS were identified as Staphylococcus epidermidis (n=20), Staphylococcus haemolyticus (n=17), Staphylococcus hominis (n=3), Staphylococcus capitis (n=1), Staphylococcus cohnii (n=1) and Staphylococcus warneri (n=1). PFGE identified one clonal lineage in S. hominis isolates and several in S. haemolyticus and S. epidermidis. Fourteen sequence types were identified in S. epidermidis, with sequence type 2 (ST2) and ST5 being predominant. Ten isolates contained SCCmec IV, seven contained SCCmec V and the others were non-typeable. ACMEs were detected in 11 S. epidermidis isolates. One S. hominis and 10 S. epidermidis isolates were icaA positive. In addition to mecA-mediated β-lactam resistance, the most frequent resistance was to gentamicin/kanamycin [aac(6')-Ie-aph(2')-Ia, aph(3')-III] (n=34), macrolides/lincosamides [erm(C), erm(A), msr, lnu(A)] (n=31), tetracycline [tet(K)] (n=22), streptomycin [str, ant(6)-Ia] (n=20), trimethoprim [dfr(A), dfr(G)] (n=17), sulfamethoxazole (n = 34) and fluoroquinolones [amino acid substitutions in GyrA and GrlA] (n=30). Clinical data suggest selection through multiple antibiotic courses and emphasize the importance of accurate diagnosis and antibiograms. MRCoNS from animal infection sites are genetically heterogeneous multidrug-resistant strains that represent a new challenge in the prevention and therapy of infections in veterinary

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.

[Oxacillin-resistant coagulase-negative Staphylococci bacteremia at a teaching hospital in Santa Maria, State of Rio Grande do Sul, Brazil].

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Rigatti, Fabiane; Tizotti, Maísa Kraulich; Hörner, Rosmari; Domingues, Vanessa Oliveira; Martini, Rosiéli; Mayer, Letícia Eichstaedt; Khun, Fábio Teixeira; de França, Chirles Araújo; da Costa, Mateus Matiuzzi

2010-01-01

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. After identification (MicroScan - Siemens), the isolates were tested for antimicrobial sensitivity using disk diffusion and automation (MicroScan - Siemens). The presence of mecA gene was identified by the polymerase chain reaction molecular technique. The most common species was Staphylococcus epidermidis (n=40, 67%). The mecA gene was detected in 54 (90%) strains, while analysis of the sensitivity profiles revealed a high rate of resistance to multiple classes of antimicrobial drugs. However, all isolates were uniformly sensitive to vancomycin and tigecycline. The cefoxitin disk was the phenotypic method that best correlated with the gold standard. Analysis of the clinical significance of CoNS isolated from hemocultures and the precise detection of oxacillin resistance represent decisive factors for the correct choice of antibiotic therapy. Although vancomycin constitutes the normal treatment in most Brazilian hospitals, reduction in its use is recommended.

Antibiotic resistance of coagulase-negative staphylococci isolated from artisanal Naples-type salami.

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Mauriello, G; Moschetti, G; Villani, F; Blaiotta, G; Coppola, S

2000-01-01

In the present paper 42 isolates from Italian salami were specified as Staphylococcus xylosus (30), Staph. capitis (1), Staph. saprophyticus (1), Staph. hominis (1), Staph. simulans (1), Staph. cohnii (1) and as Staph. spp. (7). These strains were coagulase-negative and were examined for resistance/sensitivity against 25 antibiotics including beta-lactams (7), macrolides (3), amynoglicosides (5), glycopeptides, lincosamides (4) and novobiocin, fusidic acid, chloramphenicol, rifampicin, tetracycline, minocycline. More than 64% of the strains were resistant to lincomycin, penicillin G, amoxicillin, fusidic acid and novobiocin. All the strains were multiresistant and displayed at least three resistances. Over 75% had a multiple antibiotic resistance (MAR) index between 0.2 and 0.5.

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

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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 <0.05). These results confirm the high prevalence of S. epidermidis SCCmec 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.

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

beta-Lactams and macrolides are important antibiotics for treatment of staphylococcal infections in both humans and animals. The aim of the study was to investigate the occurrence, species distribution and clonality of methicillin and erythromycin-resistant staphylococci in the nasal cavity of dogs......, 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...

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

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

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.

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

Science.gov (United States)

Boamah, Vivian Etsiapa; Agyare, Christian; Odoi, Hayford; Adu, Francis; Gbedema, Stephen Yao; Dalsgaard, Anders

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. 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 from the Brong Ahafo (48.83%), Ashanti (33.59%), and Greater Accra (17.78%) regions. Isolates from poultry litter constituted 55.47%, and farm workers 44.53%. All the isolates were susceptible to amoxicillin/clavulanic acid and amikacin. The isolates exhibited high resistance toward tetracycline (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

A study of coagulase-negative staphylococci (CoNS isolated from bovine mastitis for the presence of penicillin and methicillin resistance-encoding genes in the north west of Iran

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Dastmalchi Saei, H.

2014-11-01

Full Text Available Coagulase-negative staphylococci (CoNS are often associated with bovine mastitis and may be resistant to antimicrobial therapy. The aim of the current study was to investigate the presence of blaZ (responsible for penicillin resistance and mecA (responsible for methicillin resistance genes among 108 CoNS belonging to 9 different species isolated from bovine mastitis in seven dairy herds (H1-H7. Of 108 CoNS isolates, 44 were Staphylococcus haemolyticus, 17 S. chromogenes, 11 S. epidermidis, 11 S. warneri, 11 S. cohnii, 6 S. simulans, 4 S. hominis, 3 S. capitis, and 1 S. xylosus. The blaZ was detected in 65.7% (n=71 of all Staphylococcus spp. isolates. Five isolates were positive for the presence of mecA gene (4.6%, including 2 S. hominis, 1 S. haemolyticus, 1 S. epidermidis, and 1 S. warneri. All mecA-carrying CoNS were also positive for the blaZ gene and were recovered from two studied herds (H3 and H6. Some variations were also observed in distribution of both blaZ and mecA genes between CoNS species. This study demonstrates that CoNS from bovine mastitis can be reservoirs of blaZ gene. This study also provides evidence of the presence of methicillin resistant CoNS (MR-CoNS and emphasizes the need for their epidemiological monitoring, in order to prevent the risk of spread to human through direct contact and/or consumption of contaminated food.

Safety Evaluation of the Coagulase-Negative Staphylococci Microbiota of Salami: Superantigenic Toxin Production and Antimicrobial Resistance

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Raquel Soares Casaes Nunes

2015-01-01

Full Text Available The risks of contracting staphylococci food poisoning by the consumption of improperly manufactured salami and the possibility of this food being reservoirs for antibiotic resistance were evaluated. Nineteen coagulase-negative staphylococci (CNS strains were found in commercial and artisanal salami. The species in commercial salami were S. saprophyticus, S. sciuri, S. xylosus, and S. carnosus. Artisanal salami showed S. succinus, S. epidermidis, and S. hominis but no S. carnosus. Phylogenetic analyses grouped the strains into three major staphylococcal species groups, comprised of 4 refined clusters with similarities superior to 90%. Fifteen strains harbored multiple enterotoxin genes, with high incidence of seb/sec and sea, 57% and 50%, respectively, intermediate incidence of sed/seh/selm and sei/seln/tst-H, 33% and 27%, correspondingly, and low incidence of see/selj/selo and seg, of respectively 13% and 1%. Real time RT-PCR and enzyme-linked-immunosorbent assays confirmed the enterotoxigenicity of the strains, which expressed and produced enterotoxins in vitro. The CNS strains showed multiresistance to several antimicrobials of therapeutic importance in both human and veterinarian medicine, such as β-lactams, vancomycin, and linezolid. The effective control of undue staphylococci in fermented meat products should be adopted to prevent or limit the risk of food poisoning and the spread of antimicrobial-resistant strains.

Antimicrobial susceptibility of coagulase-negative staphylococci isolated from meat-producing ewes with mastitis

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A.M.M.P Della Libera

2010-12-01

Full Text Available Avaliou-se a sensibilidade antimicrobiana in vitro de 121 cepas de estafilococos coagulase-negativa isolada de leite de ovelhas Santa Inês, aos fármacos: penicilina, amoxicilina, ampicilina, estreptomicina, oxaciclina, neomicina, cefalotina, gentamicina e sulfonamida. A resistência à sulfonamida foi a mais frequente (27,3%, seguida pela estreptomicina (14,0% e pela oxaciclina (14,0%, enquanto da gentamicina (1,6% foi a menos frequente. Todas as cepas foram sensíveis a pelo menos um antimicrobiano, e 20,3% das cepas apresentaram resistência múltipla. Os resultados mostram a importância de Staphylococci coagulase-negativas como agentes causadores de mastite em ovinos, e o perfil de resistência múltipla indica a importância da determinação da resistência à oxaciclina como indicador da presença de ilhas de patogenicidade que contêm fatores de virulência e resistência a outros antimicrobianos que contribuem para a sobrevivência da bactéria ao tratamento.

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

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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

Prevalence and antibiotic resistance of Staphylococci isolated from ...

African Journals Online (AJOL)

, 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 ...

CRISPR-Cas Systems Features and the Gene-Reservoir Role of Coagulase-Negative Staphylococci

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Ciro C. Rossi

2017-08-01

Full Text Available The claimed role of gene reservoir of coagulase-negative staphylococci (CoNS could be contradicted by estimates that CRISPR/Cas systems are found in the genomes of 40–50% of bacteria, as these systems interfere with plasmid uptake in staphylococci. To further correlate this role with presence of CRISPR, we analyzed, by computational methods, 122 genomes from 15 species of CoNS. Only 15% of them harbored CRISPR/Cas systems, and this proportion was much lower for S. epidermidis and S. haemolyticus, the CoNS most frequently associated with opportunistic infections in humans. These systems are of type II or III, and at least two of them are located within SCCmec, a mobile genetic element of Staphylococcus bacterial species. An analysis of the spacers of these CRISPRs, which come from exogenous origin, allowed us to track the transference of the SCCmec, which was exchanged between different strains, species and hosts. Some of the spacers are derived from plasmids described in Staphylococcus species that are different from those in which the CRISPR are found, evidencing the attempt (and failure of plasmid transference between them. Based on the polymorphisms of the cas1 gene in CRISPRs of types II and III, we developed a multiplex polymerase chain reaction (PCR suitable to screen and type CRISPR systems in CoNS. The PCR was tested in 59 S. haemolyticus strains, of which only two contained a type III cas1. This gene was shown to be expressed in the exponential growth, stationary phase and during biofilm formation. The low abundance of CRISPRs in CoNS is in accordance with their role as gene reservoirs, but when present, their spacers sequence evidence and give an insight on the dynamics of horizontal genetic transfer among staphylococci.

Bacteriocins of Non-aureus Staphylococci Isolated from Bovine Milk

Science.gov (United States)

Carson, Domonique A.; Barkema, Herman W.; Naushad, Sohail

2017-01-01

ABSTRACT 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

Emergence of methicillin-resistant coagulase-negative staphylococci resistant to linezolid with rRNA gene C2190T and G2603T mutations.

Science.gov (United States)

Cidral, Thiago André; Carvalho, Maria Cícera; Figueiredo, Agnes Marie Sá; de Melo, Maria Celeste Nunes

2015-10-01

The aim of this article were to determinate the mechanism of linezolid resistance in coagulase-negative methicillin-resistant staphylococci from hospitals in the northeast of Brazil. We identified the isolates using VITEK(®) 2 and MALDI-TOF. Susceptibility to antibiotics was measured by the disk-diffusion method and by Etest(®) . Extraction of the whole genome DNA was performed, followed by screening of all the strains for the presence of mecA and cfr genes. The domain V region of 23S rRNA gene was sequenced and then aligned with a linezolid-susceptible reference strain. Pulsed-field gel electrophoresis (PFGE) macro-restriction analysis was performed. Three linezolid-resistant Staphylococcus hominis and two linezolid-resistant Staphylococcus epidermidis strains were analyzed. The isolates showed two point mutations in the V region of the 23S rRNA gene (C2190T and G2603T). We did not detect the cfr gene in any isolate by PCR. The S. hominis showed the same pulsotype, while the S. epidermidis did not present any genetic relation to each other. In conclusion, this study revealed three S. hominis and two S. epidermidis strains with resistance to linezolid due to a double mutation (C2190T and G2603T) in the domain V of the 23S rRNA gene. For the first time, the mutation of C2190T in S. epidermidis is described. This study also revealed the clonal spread of a S. hominis pulsotype between three public hospitals in the city of Natal, Brazil. These findings highlight the importance of continued vigilance of linezolid resistance in staphylococci. © 2015 APMIS. Published by John Wiley & Sons Ltd.

Identification, typing, ecology and epidemiology of coagulase negative staphylococci associated with ruminants.

Science.gov (United States)

Vanderhaeghen, Wannes; Piepers, Sofie; Leroy, Frédéric; Van Coillie, Els; Haesebrouck, Freddy; De Vliegher, Sarne

2015-01-01

Since phenotypic methods to identify coagulase negative staphylococci (CNS) from the milk of ruminants often yield unreliable results, methods for molecular identification based on gene sequencing or fingerprinting techniques have been developed. In addition to culture-based detection of isolates, culture-independent methods may be of interest. On the basis of molecular studies, the five CNS species commonly causing intramammary infections (IMI) are Staphylococcus chromogenes, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus simulans and Staphylococcus xylosus. Current knowledge suggests that S. chromogenes is a bovine-adapted species, with most cases of IMI due to this bacterium being opportunistic. S. haemolyticus also appears to be an opportunistic pathogen, but this bacterium occupies a variety of habitats, the importance of which as a source of IMI remains to be elucidated. S. xylosus appears to be a versatile species, but little is known of its epidemiology. S. epidermidis is considered to be a human-adapted species and most cases of IMI appear to arise from human sources, but the organism is capable of residing in other habitats. S. simulans typically causes contagious IMI, but opportunistic cases also occur and the ecology of this bacterium requires further study. Further studies of the ecology and epidemiology of CNS as a cause of IMI in cattle are required, along with careful attention to classification of these bacteria and the diseases they cause. Copyright © 2014 Elsevier Ltd. All rights reserved.

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

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

Characterization of Coagulase-Negative Staphylococci and pheno-genotypic beta lactam resistance evaluation in samples from bovine Intramammary infection

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D.A. Melo

Full Text Available ABSTRACT This study aimed to identify Coagulase-Negative Staphylococci (CoNS species isolated from bovine mastitis, through phenotypic and PCR-RFLP (Restriction Fragment Length Polymorphism-Polimerase Chain Reaction methods and to compare both techniques to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS technique. Among them, the PCR-RFLP method, using a partially conserved sequence of the groEL gene, is a promising alternative, because of its reproducibility and reliability. On the other hand, the proteomic technique MALDI-TOF MS provides an accurate and much faster diagnosis and has been increasingly employed in microbiological identification. The pheno-genotypic profiles of beta-lactam resistance were also investigated, this characterization is important, considering that the use of antimicrobials is a key element for mastitis control in dairy farms. The concordance of the phenotypic, PCR-RFLP and MALDI-TOF MS assays to identify CoNS species was 77,5% (31/40. S. chromogenes was the species most frequently isolated. Antibiotic resistance rate was relatively low, registering values of 25.5% to penicillin, 9.6% to oxacillin and 6.2% to cefoxitin. Resistance to imipenem, cephalotin and amoxicillin+clavulanate was not observed. The mecA gene and its variant were detected in 7.6% and 4,1% of the isolates respectively. The blaZ gene was found in 43.2% of the strains resistant to penicillin.

Staphylococcal species associated with bovine mastitis in the North West of Iran: Emerging of coagulase-negative staphylococci

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Somayyeh Hosseinzadeh

2014-06-01

Full Text Available Staphylococcal mastitis is a major and costly problem of dairy cattle all over the world. The objective of this study was to isolate and identify the main staphylococcal species causing bovine mastitis in 7 dairy herds located in 4 different areas of East and West Azerbaijan provinces, Iran. Of the 158 mastitic milk samples collected, 113 staphylococcal isolates were identified (71.5% on the basis of cultural and biochemical features as well as by genus specific PCR. Then, species level identification of staphylococcal isolates was carried out using restriction fragment length polymorphism (RFLP analysis of the gap gene (933 bp. On the basis of polymerase chain reaction-RFLP, 10 different patterns were identified. Of 113 isolates, 5 (4.4% were Staphylococcus aureus and 108 (95.6% were coagulase-negative staphylococci (CoNS. Overall, nine different species of CoNS were identified as: 44 Staphylococcus haemolyticus (40.7%, 17 Staphylococcus chromogenes (15.7%, 11 Staphylococcus epidermidis, Staphylococcus warneri and Staphylococcus cohnii each (10.2%, 6 Staphylococcus simulans (5.5%, 4 Staphylococcus hominis (3.7%, 3 Staphylococcus capitis (2.7% and 1 Staphylococcus xylosus (0.9%. S. haemolyticus, S. chromogenes and S. warneri were the only species identified from clinical mastitis. No significant difference in staphylococcal IMI was found among the studied herds and regions. This study demonstrated that CoNS, especially S. haemolyticus and S. chromogenes, were predominant and thus be considered as emerging pathogens causing mastitis in the North West of Iran. Our results also revealed that the gap PCR-RFLP was useful for identifying staphylococcal isolates derived from bovine mastitis at species level.

Methicillin resistance of airborne coagulase-negative staphylococci in homes of persons having contact with a hospital environment.

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Lis, Danuta O; Pacha, Jerzy Z; Idzik, Danuta

2009-04-01

The persons having contact with a hospital environment (hospital personnel workers and discharged patients) are highly exposed to colonization with multidrug-resistant bacteria. The aim of this study was to evaluate the airborne Staphylococcus genus features in homes in which inhabitants have had contact with the hospital environment. Airborne bacteria were collected using a 6-stage Anderson impactor. The Staphylococcus species composition and resistance to methicillin, and other antimicrobial agents among 3 coagulase-negative staphylococci (CNS) species (S cohnii spp cohnii, S epidermidis, S hominis), were determined. Antibiotic resistance of isolates was tested using the agar screen method with methicillin, the polymerase chain reaction technique to detect the mecA gene, and the disk diffusion method. A higher prevalence of methicillin-resistant (MR) strains among the species isolated (40% of S epidermidis, 40% of S hominis, and 60% of S cohnii spp cohnii) was found in homes of persons who had contact with a hospital environment compared with the reference homes (only 12% of S hominis). The mecA gene was revealed in all MR S epidermidis strains and in some MR S hominis (50%) and S cohnii spp cohnii (33%) strains. All isolated MR CNS strains were susceptible to vancomycin, rifampicin, and linezolid. High numbers of airborne multidrug-resistant MR CNS in the homes of persons having contact with a hospital environment indicates that such inhabitants pose a risk of intrafamilial spreading of MR strains via air.

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.

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.

Bacteriocins of Non-aureus Staphylococci Isolated from Bovine Milk.

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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

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

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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. Copyright © 2013 Elsevier Ltd. All rights reserved.

Cytotoxic Activity of Coagulase-Negative Staphylococci in Bovine Mastitis

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Zhang, Songlin; Maddox, Carol W.

2000-01-01

Secreted toxins play important roles in the pathogenesis of bacterial infections. In this study, we examined the presence of secreted cytotoxic factors of coagulase-negative staphylococci (CoNS) from bovine clinical and subclinical mastitis. A 34- to 36-kDa protein with cell-rounding cytotoxic activity was found in many CoNS strains, especially in Staphylococcus chromogenes strains. The protein caused cell detachment and cell rounding in several cell lines, including HEp-2, Int 407, CHO-K1, and Y-1 cells. Native protein recovered from nondenatured polyacrylamide gel electrophoresis showed both cytotoxic activity and casein hydrolysis activity. The purified protein had a pH optimal at 7.2 to 7.5 and a pI of 5.1 and was heat labile. The proteolytic activity could be inhibited by zinc and metal specific inhibitors such as 1,10-phenanthroline and EDTA, indicating that it is a metalloprotease. Protein mass analysis and peptide sequencing indicated that the protein is a novel metalloprotease. Different bacterial strains expressed variable levels of 34- to 36-kDa protease, which may provide an indication of strain virulence. PMID:10678913

Prevalence and characteristics of methicillin-resistant coagulase-negative staphylococci from livestock, chicken carcasses, bulk tank milk, minced meat, and contact persons

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Zweifel Claudio

2011-01-01

Full Text Available Abstract Background Methicillin-resistant coagulase-negative staphylococci (MR-CNS are of increasing importance to animal and public health. In veterinary medicine and along the meat and milk production line, only limited data were so far available on MR-CNS characteristics. The aim of the present study was to evaluate the prevalence of MR-CNS, to identify the detected staphylococci to species level, and to assess the antibiotic resistance profiles of isolated MR-CNS strains. Results After two-step enrichment and growth on chromogenic agar, MR-CNS were detected in 48.2% of samples from livestock and chicken carcasses, 46.4% of samples from bulk tank milk and minced meat, and 49.3% of human samples. Using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS, 414 selected MR-CNS strains belonged to seven different species (S. sciuri, 32.6%; S. fleurettii, 25.1%; S. haemolyticus, 17.4%; S. epidermidis, 14.5%, S. lentus, 9.2%; S. warneri, 0.7%; S. cohnii, 0.5%. S. sciuri and S. fleurettii thereby predominated in livestock, BTM and minced meat samples, whereas S. epidermidis and S. haemolyticus predominated in human samples. In addition to beta-lactam resistance, 33-49% of all 414 strains were resistant to certain non-beta-lactam antibiotics (ciproflaxacin, clindamycin, erythromycin, tetracycline. Conclusions A high prevalence of MR-CNS was found in livestock production. This is of concern in view of potential spread of mecA to S. aureus (MRSA. Multiresistant CNS strains might become an emerging problem for veterinary medicine. For species identification of MR-CNS isolated from different origins, MALDI-TOF MS proved to be a fast and reliable tool and is suitable for screening of large sample amounts.

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.

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

Conjunctival sac bacterial flora isolated prior to cataract surgery

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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

Nasal carriage of methicillin resistant staphylococci

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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...

Comparative evaluation of methods for the detection of biofilm formation in coagulase-negative staphylococci and correlation with antibiogram

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Shrestha LB

2018-04-01

Full Text Available Lok Bahadur Shrestha, Narayan Raj Bhattarai, Basudha Khanal Department of Microbiology and Infectious Diseases, B. P. Koirala Institute of Health Sciences, Dharan, Nepal Introduction: Coagulase-negative staphylococci (CNS are normal commensals of human skin and mucous membranes. The objective of the study was to determine the prevalence of CNS among clinical isolates, characterize them up to species level, compare the three conventional methods for detection of biofilm formation, and study their antimicrobial susceptibility pattern.Materials and methods: CNS were obtained from various clinical samples including blood, urine, central venous catheter tips, endotracheal tube aspirate, and pus during a 1-year period (July 1, 2014, to June 30, 2015. Characterization up to species level was done using biochemical tests, and biofilm formation was detected by tube adherence, Congo red agar, and tissue culture plate method. Antimicrobial susceptibility testing was performed following Clinical and Laboratory Standards Institute guidelines.Results: A total of 71 CNS isolates, comprising of seven species were obtained. Staphylococcus epidermidis was the most common species followed by S. saprophyticus and S. haemolyticus. We detected biofilm formation in 71.8% of isolates. Considering the fact that tissue culture plate method is the gold standard, sensitivity of tube adherence method and Congo red agar method was found as 82% and 78%, respectively. The isolates exhibited high resistance toward penicillin (90%, azithromycin (60%, co-trimoxazole (60%, and ceftriaxone (40%, while all were susceptible to vancomycin and linezolid. Biofilm former isolates showed higher resistance than the non-formers.Conclusion: Among 71 CNS isolated, S. epidermidis was the most common isolate followed by S. saprophyticus and S. haemolyticus. Biofilm formation was detected in 71.8% of the isolates. All of the methods were effective in detecting biofilm-producing CNS strains. The

Prevalence and Genetic Basis of Antimicrobial Resistance in Non-aureus Staphylococci Isolated from Canadian Dairy Herds

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Nobrega, Diego B.; Naushad, Sohail; Naqvi, S. Ali; Condas, Larissa A. Z.; Saini, Vineet; Kastelic, John P.; Luby, Christopher; De Buck, Jeroen; Barkema, Herman W.

2018-01-01

Emergence and spread of antimicrobial resistance is a major concern for the dairy industry worldwide. Objectives were to determine: (1) phenotypic and genotypic prevalence of drug-specific resistance for 25 species of non-aureus staphylococci, and (2) associations between presence of resistance determinants and antimicrobial resistance. Broth micro-dilution was used to determine resistance profiles for 1,702 isolates from 89 dairy herds. Additionally, 405 isolates were sequenced to screen for resistance determinants. Antimicrobial resistance was clearly species-dependent. Resistance to quinupristin/dalfopristin was common in Staphylococcus gallinarum (prevalence of 98%), whereas S. cohnii and S. arlettae were frequently resistant to erythromycin (prevalence of 63 and 100%, respectively). Prevalence of resistance was 10% against β-lactams and tetracyclines. In contrast, resistance to antimicrobials critically important for human medicine, namely vancomycin, fluoroquinolones, linezolid and daptomycin, was uncommon (< 1%). Genes encoding multidrug-resistance efflux pumps and resistance-associated residues in deducted amino acid sequences of the folP gene were the most frequent mechanisms of resistance, regardless of species. The estimated prevalence of the mecA gene was 17% for S. epidermidis. Several genes, including blaZ, mecA, fexA, erm, mphC, msrA, and tet were associated with drug-specific resistance, whereas other elements were not. There were specific residues in gyrB for all isolates of species intrinsically resistant to novobiocin. This study provided consensus protein sequences of key elements previously associated with resistance for 25 species of non-aureus staphylococci from dairy cattle. These results will be important for evaluating effects of interventions in antimicrobial use in Canadian dairy herds. PMID:29503642

Prevalence and Genetic Basis of Antimicrobial Resistance in Non-aureus Staphylococci Isolated from Canadian Dairy Herds

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Diego B. Nobrega

2018-02-01

Full Text Available Emergence and spread of antimicrobial resistance is a major concern for the dairy industry worldwide. Objectives were to determine: (1 phenotypic and genotypic prevalence of drug-specific resistance for 25 species of non-aureus staphylococci, and (2 associations between presence of resistance determinants and antimicrobial resistance. Broth micro-dilution was used to determine resistance profiles for 1,702 isolates from 89 dairy herds. Additionally, 405 isolates were sequenced to screen for resistance determinants. Antimicrobial resistance was clearly species-dependent. Resistance to quinupristin/dalfopristin was common in Staphylococcus gallinarum (prevalence of 98%, whereas S. cohnii and S. arlettae were frequently resistant to erythromycin (prevalence of 63 and 100%, respectively. Prevalence of resistance was 10% against β-lactams and tetracyclines. In contrast, resistance to antimicrobials critically important for human medicine, namely vancomycin, fluoroquinolones, linezolid and daptomycin, was uncommon (< 1%. Genes encoding multidrug-resistance efflux pumps and resistance-associated residues in deducted amino acid sequences of the folP gene were the most frequent mechanisms of resistance, regardless of species. The estimated prevalence of the mecA gene was 17% for S. epidermidis. Several genes, including blaZ, mecA, fexA, erm, mphC, msrA, and tet were associated with drug-specific resistance, whereas other elements were not. There were specific residues in gyrB for all isolates of species intrinsically resistant to novobiocin. This study provided consensus protein sequences of key elements previously associated with resistance for 25 species of non-aureus staphylococci from dairy cattle. These results will be important for evaluating effects of interventions in antimicrobial use in Canadian dairy herds.

Bacterial biofilms with emphasis on coagulase-negative staphylococci

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A Oliveira

2008-01-01

Full Text Available In addition to their capacity to attach to surfaces, various groups of microorganisms also produce an extracellular polymeric substance known as "slime". This slime forms a thin layer around cells known as biofilm. Thus, biofilm structure comprises bacterial cells and an extracellular polymeric substance. It also presents a defined architecture, providing the microorganisms with an excellent protective environment and favoring the exchange of genetic material between cells as well as intercellular communication. The ability to produce biofilm is observed in a large group of bacteria, including coagulase-negative staphylococci (CNS which are the predominant microorganisms of normal skin flora and have been implicated as the causative agents of hospital infections. Bacteremia caused by these agents is common in immunodepressed persons, in patients with cancer, in adult and neonatal intensive care units (ICU and in patients using catheters or other prosthetic devices. The pathogenicity of CNS infections is probably related to the production of slime, which adheres preferentially to plastic and smooth surfaces, forming a biofilm that protects against attacks from the immune system and against antibiotic treatment, a fact hindering the eradication of these infections. The main objective of the present review was to describe basic and genetic aspects of biofilm formation and methods for its detection, with emphasis on biofilm creation by CNS and its relationship with diseases caused by these microorganisms which are becoming increasingly more frequent in the hospital environment.

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...

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

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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.

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

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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...... usage in both animal production and in humans....... 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...

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

Short communication: Evaluation of MALDI-TOF mass spectrometry and a custom reference spectra expanded database for the identification of bovine-associated coagulase-negative staphylococci.

Science.gov (United States)

Cameron, M; Perry, J; Middleton, J R; Chaffer, M; Lewis, J; Keefe, G P

2018-01-01

This study evaluated MALDI-TOF mass spectrometry and a custom reference spectra expanded database for the identification of bovine-associated coagulase-negative staphylococci (CNS). A total of 861 CNS isolates were used in the study, covering 21 different CNS species. The majority of the isolates were previously identified by rpoB gene sequencing (n = 804) and the remainder were identified by sequencing of hsp60 (n = 56) and tuf (n = 1). The genotypic identification was considered the gold standard identification. Using a direct transfer protocol and the existing commercial database, MALDI-TOF mass spectrometry showed a typeability of 96.5% (831/861) and an accuracy of 99.2% (824/831). Using a custom reference spectra expanded database, which included an additional 13 in-house created reference spectra, isolates were identified by MALDI-TOF mass spectrometry with 99.2% (854/861) typeability and 99.4% (849/854) accuracy. Overall, MALDI-TOF mass spectrometry using the direct transfer method was shown to be a highly reliable tool for the identification of bovine-associated CNS. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

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

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

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

Identification of Coagulase-Negative Staphylococci from Bovine Intramammary Infection by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

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Gonçalves, Juliano Leonel; Barreiro, Juliana Regina; Braga, Patrícia Aparecida de Campos; Prada e Silva, Luis Felipe; Eberlin, Marcos Nogueira

2014-01-01

Coagulase-negative staphylococci (CoNS) are among the main pathogens causing bovine intramammary infection (IMI) in many countries. However, one of the limitations related to the specific diagnosis of CoNS is the lack of an accurate, rapid, and convenient method that can differentiate the bacterial species comprising this group. The aim of this study was to evaluate the ability of matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) to accurately identify CoNS species in dairy cow IMI. In addition, the study aimed to determine the frequency of CoNS species causing bovine IMI. A total of 108 bacterial isolates were diagnosed as CoNS by microbiological cultures from two milk samples collected from 21 dairy herds; the first sample was collected at the cow level (i.e., 1,242 composite samples from all quarters), while the second sample was collected at the mammary quarter level (i.e., 1,140 mammary samples collected from 285 cows). After CoNS isolation was confirmed by microbiological culture for both samples, all CoNS isolates (n = 108) were genotypically differentiated by PCR restriction fragment length polymorphism (RFLP) analysis of a partial groEL gene sequence and subjected to the MALDI-TOF MS identification procedure. MALDI-TOF MS correctly identified 103 (95.4%) of the CoNS isolates identified by PCR-RFLP at the species level. Eleven CoNS species isolated from bovine IMI were identified by PCR-RFLP, and the most prevalent species was Staphylococcus chromogenes (n = 80; 74.1%). In conclusion, MALDI-TOF MS may be a reliable alternative method for differentiating CoNS species causing bovine IMI. PMID:24622096

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

OpenAIRE

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

2016-01-01

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.

Novel Multiplex PCR Assay for Detection of Chlorhexidine-Quaternary Ammonium, Mupirocin, and Methicillin Resistance Genes, with Simultaneous Discrimination of Staphylococcus aureus from Coagulase-Negative Staphylococci.

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McClure, Jo-Ann; Zaal DeLongchamp, Johanna; Conly, John M; Zhang, Kunyan

2017-06-01

Methicillin-resistant Staphylococcus aureus (MRSA) is a clinically significant pathogen that is resistant to a wide variety of antibiotics and responsible for a large number of nosocomial infections worldwide. The Agency for Healthcare Research and Quality and the Centers for Disease Control and Prevention recently recommended the adoption of universal mupirocin-chlorhexidine decolonization of all admitted intensive care unit patients rather than MRSA screening with targeted treatments, which raises a serious concern about the selection of resistance to mupirocin and chlorhexidine in strains of staphylococci. Thus, a simple, rapid, and reliable approach is paramount in monitoring the prevalence of resistance to these agents. We developed a simple multiplex PCR assay capable of screening Staphylococcus isolates for the presence of antiseptic resistance genes for chlorhexidine and quaternary ammonium compounds, as well as mupirocin and methicillin resistance genes, while simultaneously discriminating S. aureus from coagulase-negative staphylococci (CoNS). The assay incorporates 7 PCR targets, including the Staphylococcus 16S rRNA gene (specifically detecting Staphylococcus spp.), nuc (distinguishing S. aureus from CoNS), mecA (distinguishing MRSA from methicillin-susceptible S. aureus ), mupA and mupB (identifying high-level mupirocin resistance), and qac and smr (identifying chlorhexidine and quaternary ammonium resistance). Our assay demonstrated 100% sensitivity, specificity, and accuracy in a total of 23 variant antiseptic- and/or antibiotic-resistant control strains. Further validation of our assay using 378 randomly selected and previously well-characterized local clinical isolates confirmed its feasibility and practicality. This may prove to be a useful tool for multidrug-resistant Staphylococcus monitoring in clinical laboratories, particularly in the wake of increased chlorhexidine and mupirocin treatments. Copyright © 2017 American Society for Microbiology.

Understanding the physiology and adaptation of staphylococci: a post-genomic approach.

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Becker, Karsten; Bierbaum, Gabriele; von Eiff, Christof; Engelmann, Susanne; Götz, Friedrich; Hacker, Jörg; Hecker, Michael; Peters, Georg; Rosenstein, Ralf; Ziebuhr, Wilma

2007-11-01

Staphylococcus aureus as well as coagulase-negative staphylococci are medically highly important pathogens characterized by an increasing resistance rate toward many antibiotics. Although normally being skin and mucosa commensals, some staphylococcal species and strains have the capacity to cause a wide range of infectious diseases. Many of these infections affect immunocompromised patients in hospitals. However, community-acquired staphylococcal infections due to resistant strains are also currently on the rise. In the light of this development, there is an urgent need for novel anti-staphylococcal therapeutic and prevention strategies for which a better understanding of the physiology of these bacteria is an essential prerequisite. Within the past years, staphylococci have been in the focus of genomic research, resulting in the determination and publication of a range of full-genome sequences of different staphylococcal species and strains which provided the basis for the design and application of DNA microarrays and other genomic tools. Here we summarize the results of the project group 'Staphylococci' within the research network 'Pathogenomics' giving new insights into the genome structure, molecular epidemiology, physiology, and genetic adaptation of both S. aureus and coagulase-negative staphylococci.

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

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

2016-01-01

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. PMID:26912749

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.

Rapid detection of methicillin resistance in coagulase-negative staphylococci by a penicillin-binding protein 2a-specific latex agglutination test.

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Horstkotte, M A; Knobloch, J K; Rohde, H; Mack, D

2001-10-01

The detection of PBP 2a by the MRSA-Screen latex agglutination test with 201 clinical coagulase-negative staphylococci had an initial sensitivity of 98% and a high degree of specificity for Staphylococcus epidermidis strains compared to PCR for mecA. Determination of oxacillin MICs evaluated according to the new breakpoint (0.5 microg/ml) of the National Committee for Clinical Laboratory Standards exhibited an extremely low specificity for this population.

Coagulase-negative staphylococci: pathogenesis, occurrence of antibiotic resistance genes and in vitro effects of antimicrobial agents on biofilm-growing bacteria.

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Szczuka, Ewa; Jabłońska, Lucyna; Kaznowski, Adam

2016-12-01

Coagulase-negative staphylococci (CoNS) are opportunistic pathogens that particularly cause infections in patients with implanted medical devices. The present research was performed to study the virulence potential of 53 clinical isolates of Staphylococcus capitis, Staphylococcus auricularis, Staphylococcus lugdunensis, Staphylococcus simulans, Staphylococcus cohnii and Staphylococcus caprae. All clinical strains were clonally unrelated. Isolates carried genes encoding resistance to β-lactam (mecA) (15 %), aminoglycoside [aac(6')/aph(2″)(11 %), aph (3')-IIIa (15 %), ant(4')-Ia (19 %)] and macrolide, lincosamide and streptogramin B (MLSB) [erm(A) (4 %), erm(B) (13 %), erm(C) (41 %), msr(A) (11 %)] antibiotics. CoNS isolates (64 %) were able to form biofilms. Confocal laser scanning microscopy revealed that these biofilms formed a three-dimensional structure composed mainly of living cells. All biofilm-positive strains carried the ica operon. In vitro studies demonstrated that a combination treatment with tigecycline and rifampicin was more effective against biofilms than one with ciprofloxacin and rifampicin. The minimum biofilm eradication concentration values were 0.062-0.5 µg ml-1 for tigecycline/rifampicin and 0.250-2 µg ml-1 for ciprofloxacin/rifampicin. All CoNS strains adhered to the human epithelial cell line HeLa, and more than half of the isolates were able to invade the HeLa cells, although most invaded relatively poorly. The virulence of CoNS is also attributed to their cytotoxic effects on HeLa cells. Incubation of HeLa cells with culture supernatant of the CoNS isolates resulted in cell death. The results indicate that the pathogenicity of S. capitis, S. auricularis, S. lugdunensis, S. cohnii and S. caprae is multi-factorial, involving the ability of these bacteria to adhere to human epithelial cells, form biofilms and invade and destroy human cells.

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

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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

Role of Enterotoxin-Producing Staphylococci in Zoonotic Infections in Iran

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Abdolmajid Ghasemian

2018-05-01

Full Text Available Context: Staphylococci spp, are enterotoxin-producing zoonotic agents causing a variety of infections such as mastitis in animals and wound bite infections in humans. This review was conducted to determine the prevalence of Staphylococci infections especially to uncover enterotoxin-producing species in Iran. Evidence acquisition: for this review, words of "Staphylococcus", "zoonotic", "prevalence", "animals", "human" and "Iran" were searched in the internet engines such as Google Scholar, PubMed, Google, Science Direct and so on. Patients with no history of contact with animals were also included in the study for comparison aims. Both veterinary and human coagulase positive isolates were included. Data was analyzed with Graph Pad Prism 6, meta-analysis section. A total of 20 previous studies (450 clinical samples were found. S. intermedius was the predominant isolate identified in veterinary sources. Other coagulase positive spp such as S. hycus and S. simulans were isolated with lower prevalence, but S. delphini has not been detected. Conclusion: S. intermedius was the most isolate identified in veterinary sources with potential of causing infections in humans. Other coagulase positive spp such as S. hycus and S. simulans were isolated with lower prevalence, but S. delphini was not detected.

[Resistance to heavy metals in ruminal staphylococci].

Science.gov (United States)

Lauková, A

1994-01-01

Ruminal, coagulase-negative, urease and bacteriocin-like substances producing staphylococci were screened for their heavy metal ions and antibiotics resistance. All strains tested were resistant to disodium arsenate at a minimal inhibition concentration (MIC > 5 g/l) and cadmium sulphate (MIC > 4 g/l). MIC = 50-60 mg/l was determined in eight staphylococci screened in mercury chloride resistance test (Tab. I). Silver nitrate resistance was detected in seven of the bacteria used (MIC = 40-50 mg/l). All strains were novobiocin resistant. Staphylococcus cohnii subsp. urealyticum SCU 40 was found as a strain with resistance to all heavy metal ions and 5 antibiotics (Tab. II). In addition, this strain produced bacteriocin-like substance which inhibited growth of six indicators of different origin (Tab. II). The most of staphylococci were detected as heavy metal ion polyresistant strains and antibiotic polyresistant strains producing antimicrobial substances with inhibition effects against at least one indicator of different origin. These results represent the first information on heavy metal ion resistance in ruminal bacteria. They also show relation or coresistance between heavy metal ions and antibiotics. Resulting from this study, staphylococci can be used as a bioindicator model for animal environmental studies. In addition, it can be used for specific interactions studies within the framework of ruminal bacterial ecosystem and also mainly with regard to molecular genetic studies.

Occurrence of bovine mastitis and isolation of Staphyloccocus ...

African Journals Online (AJOL)

Occurrence of bovine mastitis and isolation of Staphyloccocus species from ... the samples that were positive for CMT were found to be culture positive and out of ... S. aureus and 12.3% (12/97) were coagulase-negative staphylococci (CONS).

Rapid Detection of Methicillin Resistance in Coagulase-Negative Staphylococci by a Penicillin-Binding Protein 2a-Specific Latex Agglutination Test

OpenAIRE

Horstkotte, Matthias A.; Knobloch, Johannes K.-M.; Rohde, Holger; Mack, Dietrich

2001-01-01

The detection of PBP 2a by the MRSA-Screen latex agglutination test with 201 clinical coagulase-negative staphylococci had an initial sensitivity of 98% and a high degree of specificity for Staphylococcus epidermidis strains compared to PCR for mecA. Determination of oxacillin MICs evaluated according to the new breakpoint (0.5 μg/ml) of the National Committee for Clinical Laboratory Standards exhibited an extremely low specificity for this population.

Antibiotic resistance profile of staphylococci from clinical sources ...

African Journals Online (AJOL)

JOHN

was recorded in 2 infants while another 2 had impetigo along with urinary tract infection. Urinary tract infection with skin sepsis was recorded in 6 subjects and tetanus occurred along with septicemia in 2 subjects. Distribution of Staphylococci. Altogether, a total of 178 staphylococcal isolates were cultured from 245 samples ...

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

Nasal carriage of methicillin-resistant coagulase-negative staphylococci in healthy humans is associated with occupational pig contact in a dose-response manner.

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Li, Ling; Chen, Zhiyao; Guo, Dan; Li, Shunming; Huang, Jingya; Wang, Xiaolin; Yao, Zhenjiang; Chen, Sidong; Ye, Xiaohua

2017-09-01

This study aimed to explore the association between occupational pig contact and human methicillin-resistant coagulase-negative staphylococci (MRCoNS) carriage. We conducted a cross-sectional study of pig exposed participants and controls in Guangdong, China, using a multi-stage sampling design. Participants provided a nasal swab for MRCoNS analysis and resulting isolates were tested for antibiotic susceptibility. The dose-response relation was examined using log binomial regression or Poisson regression models. The adjusted prevalence of MRCoNS carriage in pig exposed participants was 1.67 times (95% CI: 1.32-2.11) higher than in controls. The adjusted average number of resistance to different antibiotic classes of MRCoNS isolates from pig exposed participants was 1.67 times (95% CI: 1.46-1.91) higher than those from controls. Notably, we found the frequency and duration of occupational pig contact was associated with increased prevalence and increased number of resistance to different antibiotic classes of MRCoNS in a dose-response manner. When examining these relations by MRCoNS species, there was still evidence of similar exposure-response relations. Additionally, the proportion of tetracycline-resistant and tet(M)-containing MRCoNS isolates was significantly higher in pig exposed participants than in controls. These findings suggested a potential transmission of MRCoNS from livestock to humans by occupational livestock contact, and the presence of phenotypic and genotypic tetracycline resistance may aid in the differentiation of animal origins of MRCoNS isolates. Copyright © 2017 Elsevier B.V. All rights reserved.

Characterization of staphylococci and their role in pathology of children

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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

Native Chromium Resistant Staphylococci Species from a Fly Ash ...

African Journals Online (AJOL)

Sixty-six chromium-resistant Staphylococci species belonging to S. epidermidis, S. aureus, S. saprophyticus and S. arlettae were previously isolated from a chromium-polluted Fly ash (FA) dumping site in South Africa. However the genetic mechanisms responsible for chromium resistance were not known. Polymerase chain ...

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

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

Science.gov (United States)

Singh, S; Dhawan, B; Kapil, A; Kabra, S K; Suri, A; Sreenivas, V; Das, B K

2016-01-01

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. 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. 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. 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 guide in the formulation of hospital infection

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

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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.

Antibiotic resistance profile of staphylococci from clinical sources ...

African Journals Online (AJOL)

Infants, children and the aged are among the groups most vulnerable to microbial infections more so when these microbial agents become resistant to antimicrobials. The antibiotic resistant profile of Staphylococcus aureus and selected coagulase negative staphylococci were determined by standard methods. Of the 178 ...

[Investigation of biofilm formation properties of staphylococcus isolates].

Science.gov (United States)

Öcal, Duygu Nilüfer; Dolapçı, İştar; Karahan, Zeynep Ceren; Tekeli, Alper

2017-01-01

Biofilm production is an important virulence factor which allows staphylococci to adhere to medical devices. The principal component of biofilm is a "polysaccharide intercellular adhesin (PIA)" which is composed of a beta-1,6-N-acetylglucosamine polymer synthesized by an enzyme (N-acetylglucosamine transferase) encoded by the ica operon found on the bacterial chromosome. This operon is composed of four genes (A, B, C, and D), and a transposable element IS256. In this study, we aimed to determine the biofilm production characteristics of invasive/non-invasive staphylococcus isolates and different staphylococcus species. Biofilm production of 166 staphylococci was phenotypically investigated on Congo Red Agar (CRA); the presence of icaA, icaD and IS256 genes were investigated by polymerase chain reaction (PCR). 74 of the isolates (44.6%) were identified as methicillin resistant Staphylococcus aureus (MRSA), 25 (15.1%) as methicillin sensitive S.aureus (MSSA), 25 (37.3%) as Staphylococcus hominis, 20 (12%) as S.epidermidis, ten (15%) as Staphylococcus haemolyticus, nine (13.4%) as Staphylococcus capitis, two (3%) Staphylococcus saprophyticus and one (1.5%) as Staphylococcus warnerii. Of the MRSA strains, 52 were isolated from blood and 22 from nose; all MSSA strains were isolated from nose cultures. Coagulase-negative staphylococci (CoNS) strains were composed of invasive and non-invasive strains isolated from nose, catheter tip and blood cultures from patients with catheter. Production with CRA method was found to be statistically significant in invasive isolates (paureus isolates produced biofilm on CRA (paureus when compared with CoNS. Carriage of three genes and biofilm formation capacity of invasive isolates can cause refractory infections and the importance of carriage and hospital infections of these bacteria, it is important to prevent the spread of these isolates. A combination of phenotypic and genotypic tests is recommended for the investigation of biofilm

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

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

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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. PMID:23820161

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.

Impact of Serum Vancomycin Trough Levels in the Treatment of Central Nervous System Shunt Infections Caused by Coagulase-Negative Staphylococci.

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Gibson, Ashley; Kaplan, Sheldon L; Vallejo, Jesus G

2018-04-26

Coagulase-negative staphylococci (CoNS) are a common cause of pediatric ventricular shunt infections. The Infectious Diseases Society of America recommends vancomycin serum troughs of 15-20 µg/mL when treating CoNS shunt infections in adult patients. We report a series of pediatric cases of CoNS shunt infections in which clinical cure was obtained with troughs < 15 µg/mL. These findings question the relevance of this recommendation in pediatric patients. © 2018 S. Karger AG, Basel.

An Activity of Thioacyl Derivatives of 4-Aminoquinolinium Salts towards Biofilm Producing and Planktonic Forms of Coagulase-Negative Staphylococci

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Robert D. Wojtyczka

2015-01-01

Full Text Available Microorganisms present in different environments have developed specific mechanisms of settling on various abiotic and biotic surfaces by forming a biofilm. It seems to be well justified to search for new compounds enabling biofilm reduction, which is highly resistant to antibiotics. This study was thus an initial assessment of the antibacterial activity of two new quinoline derivatives of a structure of 3-thioacyl 1-methyl 4-arylaminoquinolinium salts against coagulase-negative staphylococci (CoNS isolated from a hospital environment, in a form of both biofilms and in planktonic form. Thirty-three stains of CoNS isolated from the hospital environment (air, surfaces and seven reference strains from the ATCC collection were selected for the study. The mean MIC value for 1-methyl-3-benzoylthio-4-(4-chlorophenylaminoquinolinum chloride (4-chlorophenylamino derivative was 42.60 ± 19.91 μg/mL, and in the case of strains subjected to 1-methyl-3-benzoylthio-4-(4-fluorophenylaminoquinolinum chloride (4-fluorophenylamino derivative activity, the mean MIC value was 43.20 ± 14.30 μg/mL. The mean concentration of 4-chlorophenylamino derivative that inhibited biofilm formation was 86.18 ± 30.64 μg/mL. The mean concentration of 4-fluorophenylamino derivatives that inhibited biofilm formation was higher and amounted to 237.09 ± 160.57 μg/mL. Based on the results, both derivatives of the examined compounds exhibit high antimicrobial activity towards strains growing both in planktonic and biofilm form.

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

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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.

Genotypes, antibiotic resistance, and virulence factors of staphylococci from ready-to-eat food.

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Podkowik, Magdalena; Bystroń, Jarosław; Bania, Jacek

2012-01-01

Sixty-seven staphylococcal isolates belonging to 12 species were obtained from 70 ready-to-eat food products. Staphylococcus aureus (n=25), and Staphylococcus epidermidis (n=13) were dominant. Susceptibility to penicillin, oxacillin, tetracycline, clindamycin, gentamicin, erythromycin, ciprofloxacin, and vancomycin was determined. All investigated S. aureus isolates were resistant to at least one antibiotic, and fifteen isolates were resistant to four and more antibiotics. Thirty-eight coagulase-negative staphylococci (CNS) isolates were resistant to at least one antibiotic, and seventeen to four and more antibiotics. Fifteen CNS isolates were mecA positive, and grew in the presence of 6 μg/mL oxacillin. All S. aureus isolates were mecA-negative. Arginine catabolic mobile element (ACME) was found in seven S. epidermidis isolates. Five S. epidermidis isolates harbored ica operon, ACME and were able to form biofilm. Three of them also possessed IS256 element and were mecA-positive. The expression of icaA gene was comparable in five ica-positive S. epidermidis isolates. One of six mecA positive S. epidermidis isolates was classified as sequence type (ST)155, one as ST110, and two as ST88. Two methicillin-resistant Staphylococcus epidermis (MRSE) belonged to new STs, that is, ST362, and ST363. Enterotoxin genes were found in 92% of S. aureus isolates. No enterotoxin gene was detected in analyzed CNS population. We show that ready-to-eat products are an important source of antibiotic-resistant CNS and potentially virulent strains of S. epidermidis, including genotypes undistinguishable from hospital-adapted clones.

In vitro activity of ceftaroline against staphylococci from prosthetic joint infection.

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Park, Kyung-Hwa; Greenwood-Quaintance, Kerryl E; Patel, Robin

2016-02-01

We tested the in vitro activity of ceftaroline by Etest against staphylococci recovered from patients with prosthetic joint infection, including 97 Staphylococcus aureus isolates (36%, oxacillin resistant) and 74 Staphylococcus epidermidis isolates (74%, oxacillin resistant). Ceftaroline inhibited all staphylococci at ≤0.5 μg/mL. The ceftaroline MIC(90/50) values for methicillin-susceptible S. aureus, methicillin-susceptible S. epidermidis, methicillin-resistant S. aureus, and methicillin-resistant S. epidermidis were 0.19/0.125, 0.094/0.047, 0.5/0.38, and 0.38/0.19 μg/mL, respectively. Based on these in vitro findings, ceftaroline should be further evaluated as a potential therapeutic option for the treatment of prosthetic joint infection caused by methicillin-susceptible and methicillin-resistant S. aureus and S. epidermidis. Copyright © 2016 Elsevier Inc. All rights reserved.

Species-specific sensitivity of coagulase-negative Staphylococci to single antibiotics and their combinations.

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Szymańska, Grazyna; Szemraj, Magdalena; Szewczyk, Eligia M

2011-01-01

The activity of beta-lactam antibiotics (oxacillin, cloxacillin, cephalotin), vancomycin, gentamicin and rifampicin applied in vitro individually and in combination against 37 nosocomial methicillin-resistant strains of coagulase-negative staphylococci (CNS) was assessed to demonstrate the heterogeneity of this group of bacteria and estimate the chance of the efficacy of such therapy. The strains belonged to four species: Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus cohnii, Staphylococcus hominis. They originated from a hospital environment and from the skin of medical staff of the intensive care unit of a paediatric ward at a university hospital. All strains were methicillin-resistant, according to CLSI standards, but individual strains differed in MIC(ox) values. Susceptibility to other tested antibiotics was also characteristic for the species. The increased susceptibility to antibiotics in combinations, tested by calculating the fractional inhibitory concentration (FIC) index, concerned 26 out of 37 investigated strains and it was a feature of a particular species. Combinations of vancomycin and cephalotin against S. epidermidis and oxacillin with vancomycin were significant, as well as cephalotin and rifampicin in growth inhibition of multiresistant S. haemolyticus strains.

Coagulase-negative staphylococci from non-mastitic bovine mammary gland: characterization of Staphylococcus chromogenes and Staphylococcus haemolyticus by antibiotic susceptibility testing and pulsed-field gel electrophoresis.

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Pate, Mateja; Zdovc, Irena; Avberšek, Jana; Ocepek, Matjaž; Pengov, Andrej; Podpečan, Ožbalt

2012-05-01

During routine microbiological examination of milk samples from dairy cows without clinical signs of mastitis, quarter milk samples of 231 dairy cows from 12 herds were investigated for the presence of coagulase-negative staphylococci (CNS). The isolates were identified on the basis of colony morphology, Gram staining, catalase and coagulase test and the commercial kit, API Staph. CNS was detected in 29% (67/231) of the cows. A total of seven CNS species were identified with the most prevalent being Staphylococcus (Staph.) chromogenes (30%) and Staph. haemolyticus (28·8%), followed by Staph. simulans (11·2%), Staph. xylosus (11·2%), Staph. epidermidis (7·5%), Staph. hyicus (6·3%) and Staph. sciuri (5%). The predominant species, Staph. chromogenes and Staph. haemolyticus, were further characterized by antibiotic susceptibility testing using the agar disc diffusion method (Kirby-Bauer) and by pulsed-field gel electrophoresis (PFGE). Considerable resistance to ampicillin and penicillin was observed in both species. Isolates with identical or highly similar PFGE profiles were detected at the herd level despite a marked heterogeneity seen for both species. On the basis of somatic cell count, absence of clinical signs of inflammation and heterogeneity of genotypes, we assume that CNS isolated in this study could not be considered as important causative agents of the bovine mammary gland inflammation.

Research on the Phenotypic Characterization of Mrsa Strains Isolated from Animals

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Iulia Maria BUCUR

2017-05-01

Full Text Available Keywords: chromogen, methicillin, MRSA, resistance Introduction: Currently, both in staphylococci isolated from animals with different diseases, as well as in humans, the MRSA strains (Methicillin Resistant S. aureus are monitored, as the methicillin resistance is associated with the resistance to other antibiotic groups. Methicillin resistance is encoded by mec staphylococcal chromosomal cassettes (SCCmec, which are islands of resistance. These strains can be identified by molecular biology tests and tests that reveal several phenotypic characteristics. The research was made in order to characterize and identify phenotypically the MRSA staphylococci strains isolated from animals. Materials and Methods: Researches were made on 240 coagulase positive and coagulase negative strains of staphylococci. Mannitol fermentation was tested on Champan medium, free coagulase was revealed on Baird-Parker medium and to identify S. aureus subsp. aureus was used the chromogenic medium Chromatic Staph. Methicillin-resistant strains were detected by disc diffusion method, using biodiscs with methicillin, oxacillin and cefoxitin. Also, to identify the MRSA strains, was used the chromogenic medium Chromatic MRSA. Results: The isolates were positive to mannitol and produced complete haemolysis or were unhaemolytic. A total of 44 strains produced free coagulase on Baird-Parker medium, considered coagulase positive strains, while 196 were coagulase negative strains. The isolates conducted differently to methicillin: 22,08% of strains were resistant, 51,25% of strains were susceptible and 26,66% had intermediate resistance, while the resistant strains to oxacillin were 42,91%. The increased frequency of methicillin-resistant strains of staphylococci and, particularly, MRSA strains, determined using the cefoxitin disk diffusion test, which is more reliable than methicillin and oxacillin. On the MRSA chromogenic medium, the methicillin-resistant strains of staphylococci

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.

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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.

Biofilm synthesis and its relationship with genetic characteristics in clinical methicillin-resistant staphylococci

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Nikolaos Giormezis

2015-09-01

Full Text Available Staphylococcus aureus can cause a broad range of infections, including skin infections, pneumonia and bacteraemia. Coagulase-negative staphylococci (CNS, mainly S. epidermidis, have also emerged as important pathogens, especially in immunocompromised patients or those with prosthetic devices, such as intravascular catheters or biomaterials. Of great importance in the initiation of these infections is the ability of staphylococci to adhere to various surfaces, such as host tissues and prosthetic devices and to form biofilm. The staphylococcal adhesins are encoded by a number of genes such as fnbA (S. aureus fibronectin binding protein A, sasG (S. aureus surface protein G, aap (S. epidermidis accumulation associated protein, bhp (Bap homologue protein and fbe (fibrinogen binding protein epidermidis. In this study, 106 methicillin-resistant S. aureus (MRSA, 145 methicillin-resistant S. epidermidis (MRSE and 70 non-epidermidis methicillin-resistant CNS (MR-CNS; 58 S. haemolyticus, 10 S. hominis and two S. lugdunensis were compared in terms of biofilm formation, antimicrobial resistance, clonal distribution and adhesin genes carriage. Isolates were classified into pulsotypes by PFGE and assigned to sequence types by MLST. In total, 121/321 isolates (37.7% produced biofilm and 219 (68.2% carried ica operon. The majority was multidrug resistant (94.7% and carried one or more adhesin genes. MRSE and all other MR-CNS prevailed in biofilm formation (P < 0.001 and antimicrobial resistance (P < 0.05 as compared to MRSA. MRSE also prevailed in ica carriage compared to the other methicillin-resistant staphylococci (P ≤ 0.007 Among MRSE, isolates from bacteraemias prevailed in biofilm formation (P = 0.031, whereas, strains from prosthetic device-associated infections carried more frequently aap (P = 0.003. Even though PFGE showed genetic diversity among MRSE, MLST revealed three major clones (ST2, ST5, ST16. MRSA isolates were less diverse, with five PFGE

Adherence of Staphylococci to plastic, mesothelial cells and mesothelial extracellular matrix

NARCIS (Netherlands)

Betjes, M. G.; Tuk, C. W.; Struijk, D. G.; Krediet, R. T.; Arisz, L.; Beelen, R. H.

1992-01-01

In this study we have investigated whether mesothelial cells (MC) and mesothelial extracellular matrix (ECM) are suitable substrates for the adherence of Staphylococci. Mesothelial cells were isolated from the peritoneal dialysis effluent by making use of their lack of Fc-receptors and capacity to

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

OpenAIRE

BASSO, ANA P.; MARTINS, PAULA D.; NACHTIGALL, GISELE; SAND, SUELI VAN DER; MOURA, TIANE M. DE; FRAZZON, ANA PAULA G.

2014-01-01

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...

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

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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.

Incidence of enterotoxigenic staphylococci and their toxins in foods.

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Soriano, J M; Font, G; Rico, H; Moltó, J C; Mañes, J

2002-05-01

Of 504 food samples collected from cafeterias, 19 (3.8%) yielded strains of enterotoxigenic staphylococci, and 10 (52.6%), 4 (21.1%), 3 (15.8%), and 2 (10.5%) of these strains produced enterotoxins C (SEC), D (SED), B (SEB), and A (SEA), respectively. Moreover, SEA, SEB, and SEC were isolated from three hamburger samples. Of 181 food samples collected from four restaurants before the implementation of the hazard analysis and critical control point (HACCP) system, 7 (3.9%) were found to contain enterotoxigenic strains, and SED, SEC, and SEA were produced by 4 (57.1%), 2 (28.6%), and 1 (14.3%) of these strains, respectively. One meatball sample with SEC was detected in a restaurant. After the implementation of the HACCP system in four restaurants, neither enterotoxigenic staphylococci nor enterotoxins were detected in 196 studied samples.

The Staphylococci Phages Family: An Overview

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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.

Rapid identification of staphylococci by Raman spectroscopy

Czech Academy of Sciences Publication Activity Database

Rebrošová, K.; Šiler, Martin; Samek, Ota; Růžička, F.; Bernatová, Silvie; Holá, V.; Ježek, Jan; Zemánek, Pavel; Sokolová, J.; Petráš, P.

2017-01-01

Roč. 7, NOV (2017), s. 1-8, č. článku 14846. ISSN 2045-2322 R&D Projects: GA ČR(CZ) GA15-20645S; GA MŠk(CZ) LO1212; GA MŠk ED0017/01/01 Institutional support: RVO:68081731 Keywords : coagulase-negative staphylococci * Raman spectroscopy * rapid identification Subject RIV: BH - Optics, Masers, Lasers OBOR OECD: Optics (including laser optics and quantum optics) Impact factor: 4.259, year: 2016

Trends of Bacterial Keratitis Culture Isolates in Jerusalem; a 13- Years Analysis.

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Michael Politis

Full Text Available 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.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.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 = <0.001. None of the pathogens had developed any resistance to Vancomycin. (P = 0.88.Coagulase negative staphylococci were the predominant bacteria isolated from patients with 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.

Intramammary infection with coagulase-negative staphylococci at parturition: Species-specific prevalence, risk factors, and effect on udder health.

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De Visscher, A; Piepers, S; Haesebrouck, F; De Vliegher, S

2016-08-01

Coagulase-negative staphylococci (CNS) are the main cause of bovine intramammary infections (IMI) in many countries. Despite a high prevalence of CNS IMI at parturition, species-specific risk factor studies, relying on accurate identification methods, are lacking. Therefore, this observational study aimed at determining the prevalence and distribution of different CNS species causing IMI in fresh heifers and dairy cows in Flemish dairy herds and identifying associated species- and subgroup-specific risk factors at the herd, cow, and quarter level. The effect on udder health was investigated as well. Staphylococcus chromogenes, S. sciuri, and S. cohnii were the most frequently isolated species. The only CNS species causing IMI in fresh heifers and dairy cows in all herds was Staphylococcus chromogenes, whereas large between-herd differences in distribution were observed for the other species. Quarters from heifers and quarters with an inverted teat end had higher odds of being infected with S. chromogenes, S. simulans, or S. xylosus as well as with S. chromogenes solely. Prepartum teat apex colonization with S. chromogenes increased the likelihood of S. chromogenes IMI in the corresponding quarters at parturition. Quarters with dirty teat apices before calving were more likely to be infected with S. cohnii, S. equorum, S. saprophyticus, or S. sciuri, supporting the environmental nature of these CNS species. Three species (S. chromogenes, S. simulans, and S. xylosus) were associated with a higher quarter somatic cell count at parturition as compared with uninfected quarters. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Mutasynthesis of lincomycin derivatives with activity against drug-resistant staphylococci

Czech Academy of Sciences Publication Activity Database

Ulanová, Dana; Novotná, Jitka; Smutná, Yvona; Kameník, Zdeněk; Gažák, Radek; Šulc, Miroslav; Sedmera, Petr; Kadlčík, Stanislav; Plháčková, Kamila; Janata, Jiří

2010-01-01

Roč. 54, č. 2 (2010), s. 927-930 ISSN 0066-4804 R&D Projects: GA MŠk 2B08064; GA AV ČR IAA500200810 Institutional research plan: CEZ:AV0Z50200510 Keywords : COAGULASE-NEGATIVE STAPHYLOCOCCI * ERYTHROMYCIN RESISTANCE * SUBSTRATE-SPECIFICITY Subject RIV: EE - Microbiology, Virology Impact factor: 4.672, year: 2010

Genetic regulation of the intercellular adhesion locus in staphylococci

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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.

Risk Factors for Nasal Colonization by Methicillin-Resistant Staphylococci in Healthy Humans in Professional Daily Contact with Companion Animals in Portugal.

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Rodrigues, Ana Catarina; Belas, Adriana; Marques, Cátia; Cruz, Luís; Gama, Luís T; Pomba, Constança

2018-05-01

Methicillin-resistant staphylococci (MRS), namely Staphylococcus aureus (MRSA) and Staphylococcus pseudintermedius (MRSP), are opportunistic agents of great importance in human and veterinary medicine. The aims of this study were to investigate the frequency, persistence, and risk factors associated with nasal colonization by MRS in people in daily contact with animals in Portugal. Seventy-nine out of 129 (61.2%) participants were found to be colonized by, at least, one methicillin-resistant (MR) staphylococci species (MR Staphylococcus epidermidis [n = 68], MRSA [n = 19], MR Staphylococcus haemolyticus [n = 7], MRSP [n = 2], and other coagulase-negative staphylococci [n = 4]). Three lineages were identified among the MRSA isolates (n = 7): the major human healthcare clone in Portugal (ST22-t032-IV, n = 3), the livestock-associated MRSA (ST398-t108-V, n = 3), and the New York-/Japan-related clone (ST105-t002-II, n = 1). MRSP isolates belonged to the European clone ST71-II-III. We identified two risk factors for nasal colonization by MRS in healthy humans: (i) being a veterinary professional (veterinarian and veterinary nurse) (p < 0.0001, odds ratio [OR] = 6.369, 95% confidence interval [CI, 2.683-15.122]) and (ii) have contacted with one MRSA- or MRSP-positive animal (p = 0.0361, OR = 2.742, 95% CI [1.067-7.045]). The follow-up study revealed that the majority (85%) remain colonized. This study shows that MRS in veterinary clinical practice is a professional hazard and highlights the need to implement preventive measures to minimize spread.

Distribution of coagulase-positive staphylococci in humans and dogs

OpenAIRE

Jurate Sleiniute; Jurate Siugzdaite

2015-01-01

Staphylococci form part of the normal flora of humans and a wide variety of animals. Some staphylococcal species also cause infections for human and household pets. The aim of this study was to investigate the prevalence of coagulase-positive staphylococci in humans and dogs within a common household. Coagulase-positive staphylococci could not be identified phenotypically in all cases. Molecular methods were used for the correct identification of pathogenic staphylococci. The prevalence of St...

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.

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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.

Antimicrobial susceptibility of Staphylococcus saprophyticus and urethral staphylococci.

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Marrie, T J; Kwan, C

1982-01-01

The activity of eight antimicrobial agents was determined against 115 isolates of Staphylococcus saprophyticus. All were susceptible to ampicillin, cephalexin, and trimethoprim-sulfamethoxazole and resistant to nalidixic acid and novobiocin. A bimodal pattern of susceptibility to erythromycin was observed: 80% were inhibited by 0.25 microgram/ml, whereas 13% required greater than or equal to 128 micrograms/ml. The following urethral staphylococci were susceptible to ampicillin, cephalexin, and nitrofurantoin but resistant to nalidixic acid: S. epidermidis, S. hominis, S. haemolyticus, S. warneri, S. simulans, and S. cohnii. PMID:6982679

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

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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.

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

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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

Biotechnological Methods for Precise Diagnosis of Methicillin Resistance in Staphylococci

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Aija Zilevica

2005-04-01

Full Text Available Antimicrobial resistance is one of the most urgent problems in medicine nowadays. The purpose of the study was to investigate the microorganisms resistant to first-line antimicrobials, including gram-positive cocci, particularly the methicillin-resistant Staphylococcus aureus and coagulase-negative Staphylococci, the major agents of nosocomial infections. Owing to the multi-resistance of these agents, precise diagnosis of the methicillin resistance of Staphylococci is of greatest clinical importance. It is not enough to use only conventional microbiological diagnostic methods. Biotechnological methods should be also involved. In our studies, the following methicillin resistance identification methods were used: the disk diffusion method, detection of the mecA gene by PCR, E-test and Slidex MRSA test. For molecular typing, PFGL, RAPD tests and detection of the coa gene were used. All the MRS strains were multiresistant to antibacterials. No vancomycine resistance was registered.

Antimicrobial resistance in coagulase-positive staphylococci isolated from companion animals in Australia: A one year study.

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Saputra, Sugiyono; Jordan, David; Worthing, Kate A; Norris, Jacqueline M; Wong, Hui S; Abraham, Rebecca; Trott, Darren J; Abraham, Sam

2017-01-01

Methicillin-resistant coagulase-positive staphylococci (CoPS) have become increasingly recognised as opportunistic pathogens that limit therapeutic options in companion animals. The frequency of methicillin resistance amongst clinical isolates on an Australia-wide level is unknown. This study determined antimicrobial susceptibility patterns for CoPS isolated from clinical infections in companion animals (dogs, cats and horses) as part of the first nation-wide survey on antimicrobial resistance in animal pathogens in Australia for a one-year period (January 2013 to January 2014). Clinical Staphylococcus spp. isolates (n = 888) obtained from 22 veterinary diagnostic laboratories were identified by MALDI-TOF mass spectrometry and subjected to antimicrobial susceptibility testing for 16 antimicrobials, representing 12 antimicrobial classes. Potential risk factors associated with methicillin resistance in Staphylococcus pseudintermedius isolates from dogs were analysed based on demographic factors and clinical history, including gender, age, previous antimicrobial treatment, chronic and/or recurrent diseases and site of infections. The most commonly identified CoPS were S. pseudintermedius (70.8%; dogs n = 616, cats n = 13) and S. aureus (13.2%, horses n = 53, dogs n = 47 and cats n = 17). Overall, the frequency of methicillin resistance among S. pseudintermedius (MRSP) and S. aureus (MRSA) was 11.8% and 12.8%, respectively. MRSP isolates were strongly associated with resistance to fluoroquinolones (OR 287; 95%CI 91.2-1144.8) and clindamycin (OR 105.2, 95%CI 48.5-231.9). MRSA isolates from dogs and cats were also more likely to be resistant to fluoroquinolones (OR 5.4, 95%CI 0.6-252.1), whereas MRSA from horses were more likely to be resistant to rifampicin. In multivariate analysis, MRSP-positive status was significantly associated with particular infection sites, including surgical (OR 8.8; 95%CI 3.74-20.7), and skin and soft tissue (OR 3.9; 95%CI 1.97-7.51). S

Bacteriologic Profile and Antibiogram of Blood Culture Isolates from a Children's Hospital in Kabul

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Tariq, O. M.

2014-01-01

Objective: To identify the bacterial pathogens causing paediatric septicaemia in Kabul and to determine their antibiogram to improve empirical antibiotic therapy. Study Design: Cross-sectional study. Place and Duration of Study: Microbiology Laboratory of FMIC, Kabul, Afghanistan, from January 2010 to June 2012. Methodology: Blood cultures from suspected cases of sepsis were processed in BD (Becton Dickinson, USA) for culture BACTEC 9240 Blood Culture System. Positive growths were examined and isolates were identified by conventional biochemical tests. Bacteria were identified to the species level using various Analytical Profile Index (API) identification strips. Antibiotic susceptibility testing was performed by modified Kirby-Bauer disk diffusion method. Drug resistant strains were studied for extended spectrum beta lactamase (ESBL) production by combination disk method and for methicillin resistant Staphylococcus aureus (MRSA) by Cefoxitin disk diffusion method. Results: Out of a total 3360 blood cultures received from in-patients, 410 yielded monomicrobial growth; hence the frequency of positive blood culture was 12.2%. Out of a total 410 isolates, 212 (51.71%) were gram-negative bacilli and 184 (44.88%) were gram-positive cocci. In addition, 14 (3.41%) Candida species were also isolated. The frequently isolated species of gram-negative bacteria belonged to Enterobacteriaceae and included 66 Klebsiella (16.1%), 42 Enterobacter (10.2%), 35 Escherichia (E.) coli (8.5%) and 16 Serratia (3.9%) species. In addition, 21 (5.12%) Pseudomonas species were also isolated. Correspondingly, amongst gram-positive cocci, the most frequently isolated species were 108 coagulase-negative Staphylococci (26.34%) followed by 49 Staphylococcus aureus (11.95%) and 21 Streptococcus species (5.12%). Among gram-negative isolates, those that produced ESBL i.e., 110 out of 212 (51.9%) were found to be multidrug-resistant and showed high resistance to commonly used antibiotics namely

Bacteriologic profile and antibiogram of blood culture isolates from a children's hospital in Kabul.

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Tariq, Tariq Mahmud

2014-06-01

To identify the bacterial pathogens causing paediatric septicaemia in Kabul and to determine their antibiogram to improve empirical antibiotic therapy. Cross-sectional study. Microbiology Laboratory of FMIC, Kabul, Afghanistan, from January 2010 to June 2012. Blood cultures from suspected cases of sepsis were processed in BD (Becton Dickinson, USA) for culture BACTEC™ 9240 Blood Culture System. Positive growths were examined and isolates were identified by conventional biochemical tests. Bacteria were identified to the species level using various Analytical Profile Index (API) identification strips. Antibiotic susceptibility testing was performed by modified Kirby-Bauer disk diffusion method. Drug resistant strains were studied for extended spectrum beta lactamase (ESBL) production by combination disk method and for methicillin resistant Staphylococcus aureus (MRSA) by Cefoxitin disk diffusion method. Out of a total 3360 blood cultures received from in-patients, 410 yielded monomicrobial growth; hence the frequency of positive blood culture was 12.2%. Out of a total 410 isolates, 212 (51.71%) were gram-negative bacilli and 184 (44.88%) were gram-positive cocci. In addition, 14 (3.41%) Candida species were also isolated. The frequently isolated species of gram-negative bacteria belonged to Enterobacteriaceae and included 66 Klebsiella (16.1%), 42 Enterobacter (10.2%), 35 Escherichia (E.) coli (8.5%) and 16 Serratia (3.9%) species. In addition, 21 (5.12%) Pseudomonas species were also isolated. Correspondingly, amongst gram-positive cocci, the most frequently isolated species were 108 coagulase-negative Staphylococci (26.34%) followed by 49 Staphylococcus aureus (11.95%) and 21 Streptococcus species (5.12%). Among gram-negative isolates, those that produced ESBL i.e., 110 out of 212 (51.9%) were found to be multidrug-resistant and showed high resistance to commonly used antibiotics namely Ampicillin, Gentamicin, 3rd generation Cephalosporins, Fluoroquinolones and

The importance of staphylococci and threshold value of somatic cell count for diagnosis of sub-clinical mastitis in Pirlak sheep at mid-lactation.

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Ozenc, E; Seker, E; Baki Acar, D; Birdane, M K; Darbaz, I; Dogan, N

2011-12-01

This study investigated the bacterial agents causing sub-clinical mastitis and the mean somatic cell counts (SCC) of milk in Pirlak sheep at mid-lactation. The percentage of infected udder halves was 11.4% (53/464). The most frequently isolated species were coagulase-negative staphylococci (CNS) (64.2%), followed by Staphylococcus aureus (24.5%) and Escherichia coli (11.3%). Among the CNS, the most common species was Staphylococcus epidermidis (38.2%). The other species isolated from milk samples were Staphylococcus xylosus (17.7%), Staphylococcus chromogenes (14.7%), Staphylococcus simulans (8.8%) and Staphylococcus hyicus (8.8%). The mean SCC for culture positive and negative samples was 1742×10(3) and 161×10(3) cells/ml, respectively. A significant difference (pmastitis in Pirlak sheep. This is the first study to describe the bacterial agents causing sub-clinical mastitis and threshold limit for SCC in Pirlak sheep in Turkey. © 2011 Blackwell Verlag GmbH.

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

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Schwab, Frank; Gastmeier, Petra; Meyer, Elisabeth

2014-01-01

We investigated the relationship between average monthly temperature and the most common clinical pathogens causing infections in intensive care patients. 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. 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%. 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 host susceptibility to different bacterial pathogens

[In vitro activity of ertapenem against clinical bacterial isolates in 69 Spanish medical centers (E-test study)].

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Gobernado, M; Sanz-Rodríguez, C; Villanueva, R; Torroba, L; Redondo, E; González-Esteban, J

2007-12-01

This study was conducted to assess the in vitro activity of ertapenem against clinical bacterial isolates from patients with community-acquired intra-abdominal and lower tract respiratory infections in Spain in 2003. As the study was conducted before the marketing of ertapenem, it was also useful to define a baseline susceptibility pattern for ertapenem in each of the participating hospitals for later surveillance studies. Each partipating site identified a variable number of aerobic and facultative bacteria isolated from patients with community-acquired intra-abdominal infection or pneumonia using standard procedures. E-test strips were used for determining the minimum inhibitory concentration (MIC) of ertapenem, while for other antimicrobials either quantitative dilution techniques or qualitative diffusion procedures were used according to each microbiology laboratory's routine practice. MIC breakpoints for categorization of susceptibility provided by the CLSI were used for interpreting MIC values. A total of 2,901 recent clinical isolates from patients with community-acquired intra-abdominal infection or pneumonia hospitalized in 69 Spanish medical centers were tested. These isolates included 2,039 Gram-negative bacteria (1,646 Enterobacteriaceae, 216 Haemophilus, 123 non-fermenting Gram-negative bacteria [NFGNB] and 54 others) and 862 Gram-positive bacteria (556 pneumococci, 159 staphylococci, 96 streptococci other than S. pneumoniae, 44 enterococci and 7 others). Ertapenem was very active in vitro against Enterobacteriaceae (99.8% susceptible), Haemophilus (96.3% susceptible), pneumococci (99.6% susceptible, of which 31% were penicillin non-susceptible strains), streptococci other than S. pneumoniae (99.0% susceptible) and methicillin-susceptible staphylococci (94.8% susceptible). For other Gram-positive and Gram-negative pathogens for which ertapenem susceptible breakpoints have not been defined, MIC(90) values were 0.38 and 0.064 mg/l, respectively. As

Activity of vancomycin, linezolid, and daptomycin against staphylococci and enterococci isolated in 5 Greek hospitals during a 5-year period (2008-2012).

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Papadimitriou-Olivgeris, Matthaios; Kolonitsiou, Fevronia; Zerva, Loukia; Lebessi, Evangelia; Koutsia, Chryssa; Drougka, Eleanna; Sarrou, Styliani; Giormezis, Nikolaos; Vourli, Sofia; Doudoulakakis, Anastassios; Konsolakis, Christos; Marangos, Markos; Anastassiou, Evangelos D; Petinaki, Efthimia; Spiliopoulou, Iris

2015-12-01

The tendency of vancomycin, linezolid, and daptomycin MICs was investigated among 6920 staphylococci and enterococci during a 5-year period. Antimicrobial consumption was determined. Decrease of vancomycin MIC was detected associated with reduction in consumption. Linezolid and daptomycin remained active. An upward trend of linezolid MIC for methicillin-resistant staphylococci was observed. Copyright © 2015 Elsevier Inc. All rights reserved.

Microbiological and molecular characterization of human clinical isolates of Staphylococcus cohnii, Staphylococcus hominis, and Staphylococcus sciuri.

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Garza-González, Elvira; Morfin-Otero, Rayo; Martínez-Vázquez, Manuel A; Gonzalez-Diaz, Esteban; González-Santiago, Omar; Rodríguez-Noriega, Eduardo

2011-12-01

The incidence of coagulase-negative staphylococci reported as causative agents of nosocomial infections has risen in the last decade. The aim of this study was to characterize biofilm formation, antibiotic resistance, SCCmec type, and genetic relatedness in clinical isolates of Staphylococcus cohnii, Staphylococcus hominis, and Staphylococcus sciuri recovered from humans. Clinically relevant isolates of S. cohnii (n = 15), S. hominis (n = 9), and S. sciuri (n = 6), were collected from patients. Biofilm formation was evaluated using crystal violet staining, drug susceptibility was assessed using the broth microdilution method, and methicillin resistance was measured using the cefoxitin disk test. SCCmec was typed using 2 different methodologies, and genetic relatedness was determined by pulsed-field gel electrophoresis (PFGE). Sixty percent (9/15) of S. cohnii, 33% (3/9) of S. hominis, and 50% (3/6) of S. sciuri isolates were categorized as weak producers of biofilm. None of the isolates were resistant to vancomycin or linezolid. All 3 species showed a high resistance (> 66%) to ampicillin, levofloxacin, erythromycin, and ceftriaxone, and the majority of the isolates were methicillin-resistant. PFGE revealed that the S. cohnii isolates comprised 1 dominant clone. The S. cohnii, S. hominis, and S. sciuri isolates analyzed in this study showed a high methicillin resistance and resistance to other antimicrobials. The results of this study strongly suggest that coagulase-negative staphylococci harbour new SCCmec elements. We report the first case of a clone of S. cohnii associated with human disease.

Investigation of Linezolid Resistance in Staphylococci and Enterococci

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Gallegos, Michael; Alspaugh, Debbie

2016-01-01

The objective of this study was to investigate an apparent increase in linezolid-nonsusceptible staphylococci and enterococci following a laboratory change in antimicrobial susceptibility testing from disk diffusion to an automated susceptibility testing system. Isolates with nonsusceptible results (n = 27) from Vitek2 were subjected to a battery of confirmatory testing which included disk diffusion, Microscan broth microdilution, Clinical and Laboratory Standards Institute (CLSI) reference broth microdilution, gradient diffusion (Etest), 23S rRNA gene sequencing, and cfr PCR. Our results show that there is poor correlation between methods and that only 70 to 75% of isolates were confirmed as linezolid resistant with alternative phenotypic testing methods (disk diffusion, Microscan broth microdilution, CLSI broth microdilution, and Etest). 23S rRNA gene sequencing identified mutations previously associated with linezolid resistance in 16 (59.3%) isolates, and the cfr gene was detected in 3 (11.1%) isolates. Mutations located at positions 2576 and 2534 of the 23S rRNA gene were most common. In addition, two previously undescribed variants (at positions 2083 and 2345 of the 23S rRNA gene) were also identified and may contribute to linezolid resistance. PMID:26935728

Antimicrobial resistance in coagulase-positive staphylococci isolated from companion animals in Australia: A one year study.

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Sugiyono Saputra

Full Text Available Methicillin-resistant coagulase-positive staphylococci (CoPS have become increasingly recognised as opportunistic pathogens that limit therapeutic options in companion animals. The frequency of methicillin resistance amongst clinical isolates on an Australia-wide level is unknown. This study determined antimicrobial susceptibility patterns for CoPS isolated from clinical infections in companion animals (dogs, cats and horses as part of the first nation-wide survey on antimicrobial resistance in animal pathogens in Australia for a one-year period (January 2013 to January 2014. Clinical Staphylococcus spp. isolates (n = 888 obtained from 22 veterinary diagnostic laboratories were identified by MALDI-TOF mass spectrometry and subjected to antimicrobial susceptibility testing for 16 antimicrobials, representing 12 antimicrobial classes. Potential risk factors associated with methicillin resistance in Staphylococcus pseudintermedius isolates from dogs were analysed based on demographic factors and clinical history, including gender, age, previous antimicrobial treatment, chronic and/or recurrent diseases and site of infections. The most commonly identified CoPS were S. pseudintermedius (70.8%; dogs n = 616, cats n = 13 and S. aureus (13.2%, horses n = 53, dogs n = 47 and cats n = 17. Overall, the frequency of methicillin resistance among S. pseudintermedius (MRSP and S. aureus (MRSA was 11.8% and 12.8%, respectively. MRSP isolates were strongly associated with resistance to fluoroquinolones (OR 287; 95%CI 91.2-1144.8 and clindamycin (OR 105.2, 95%CI 48.5-231.9. MRSA isolates from dogs and cats were also more likely to be resistant to fluoroquinolones (OR 5.4, 95%CI 0.6-252.1, whereas MRSA from horses were more likely to be resistant to rifampicin. In multivariate analysis, MRSP-positive status was significantly associated with particular infection sites, including surgical (OR 8.8; 95%CI 3.74-20.7, and skin and soft tissue (OR 3.9; 95%CI 1.97-7.51. S

Antibiotic resistance patterns of coagulase-negative staphylococcus strains isolated from blood cultures of septicemic patients in Turkey.

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Koksal, F; Yasar, H; Samasti, M

2009-01-01

The aim of this study is to determine antibiotic resistance patterns and slime production characteristics of coagulase-negative Staphylococci (CoNS) caused nosocomial bacteremia. A total of 200 CoNS strains were isolated from blood samples of patients with true bacteremia who were hospitalized in intensive care units and in other departments of Istanbul University Cerrahpasa Medical Hospital between 1999 and 2006. Among 200 CoNS isolates, Staphylococcus epidermidis was the most prevalent species (87) followed by Staphylococcus haemolyticus (23), Staphylococcus hominis (19), Staphylococcus lugdunensis (18), Staphylococcus capitis (15), Staphylococcus xylosus (10), Staphylococcus warneri (8), Staphylococcus saprophyticus (5), Staphylococcus lentus (5), Staphylococcus simulans (4), Staphylococcus chromogenes (3), Staphylococcus cohnii (1), Staphylococcus schleiferi (1), and Staphylococcus auricularis (1). Resistance to methicillin was detected in 67.5% of CoNS isolates. Methicillin-resistant CoNS strains were determined to be more resistant to antibiotics than methicillin-susceptible CoNS strains. Resistance rates of methicillin-resistant and methicillin-susceptible CoNS strains to the antibacterial agents, respectively, were as follows: gentamicin 90% and 17%, erythromycin 80% and 37%, clindamycin 72% and 18%, trimethoprim-sulfamethoxazole 68% and 38%, ciprofloxacin 67% and 23%, tetracycline 60% and 45%, chloramphenicol 56% and 13% and fusidic acid 25% and 15%. None of the strains were resistant to vancomycin and teicoplanin. Slime production was detected in 86 of 200 CoNS strains. Resistance to methicillin was found in 81% of slime-positive and in 57% of slime-negative strains. Our results indicated that there is a high level of resistance to widely used agents in causative methicillin-resistant CoNS strains. However fusidic acid has the smallest resistance ratio, with the exception of glycopeptides. Additionally, most S. epidermidis strains were slime

Characterization of Staphylococcus species isolated from raw milk and milk products (lben and jben) in North Morocco.

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Bendahou, Abdrezzak; Lebbadi, Mariam; Ennanei, Latifa; Essadqui, Fatima Z; Abid, Mohammed

2008-06-01

To investigate the incidence and antibiotic resistance of staphylococcal strains isolated from milk and milk products and to trace the ecological origin of the Staphylococcus aureus isolated. Eighty-one samples of raw milk, lben (whey) and jben (cheese) were analyzed for the presence of staphylococcal strains. Isolates were identified by Gram stains, tests for coagulase, the API staph system and the WalkAway 40/96, which also determines the antimicrobial susceptibility profiles. The S. aureus strains were biotyped, and variable regions of the coagulase gene were amplified using the polymerase chain reaction. The identification results showed a predominance of coagulase-negative staphylococci (54 %). Coagulase-positive staphylococci that were identified were divided into 3 groups comprising S. aureus (40%), Staphylococcus intermedius (2 %) and Staphylococcus hyicus (4%). Among the S. aureus that was isolated, biotype C was the predominant biotype. Among 40 coagulase gene PCR-amplification products, 37 produced a single band, while 3 isolates produced two bands. The antimicrobial susceptibility-profile of the staphylococcal strains revealed a high incidence of S. aureus to penicillin G. In addition, Staphylococcus lentus presented considerable resistance to the oxacillin, erythromycin and lincomycin. The presence of staphylococci in raw milk, lben and jben in areas of northern Morocco poses a health hazard, so it is necessary for the public health inspectors to properly examine the conditions during production, storage and commercialization of all products made with unpasteurized milk.

Ultrastructural changes in biofilm forms of staphylococci cultivated in a mixed culture with lactobacilli

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G. Lavryk

2017-02-01

Full Text Available The capacity of opportunistic bacteria for biofilm formation plays an important role in the development of chronic inflammatory processes, which are difficult to treat. To improve antimicrobial therapy methods, the influence of lactobacilli on the ultrastructure of biofilm-forming clinical strains of staphylococci when co-cultured was investigated. 5 biofilm-forming clinical strains of S. aureus from the skin of acne vulgaris patients (n = 24 were isolated. Using transmission electron microscopy (TEM the morphological changes of S. aureus cells in the mixed culture with standard strains of Lactobacillus plantarum 8P-A3 and clinical strains of L. fermentum (n = 4 were studied. It was found that in 48 hours after the inoculation on the medium of samples of mixed cultures of L. plantarum 8P-A3 and S. aureus growth of staphylococci was not revealed. Only in some cases of mixed cultures of L. fermentum and biofilm-forming staphylococci was growth of S. aureus obtained. In electron diffraction patterns of control samples of 24-hour staphylococcal monocultures and 48-hour lactobacilli monocultures, natural development of the population at the cellular level was observed. Destructive changes under the influence of lactobacilli (probiotic and clinical strains were detected in all ultrathin sections of the cells of biofilm-forming and planktonic staphylococci. Significant destructive changes in the cell wall of the staphylococci were observed: thickening, obtaining of irregular form, detachment of the cytoplasmic membrane, the complete destruction of the peptidoglycan layer and the emergence of "shadow cells". On all electron diffraction patterns fibrillar-threadlike structures of DNA could not be observed, but in some cases mesosome-like formations were poorly contrasted. It was established that the surface S-layer of lactobacilli was expressed on a significantly larger scale in the mixed culture with staphylococci. In mixed culture of clinical strains

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.

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

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

2016-01-01

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

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

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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

[Left-sided native valve endocarditis by coagulase-negative staphylococci: an emerging disease].

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Haro, Juan Luis; Lomas, José M; Plata, Antonio; Ruiz, Josefa; Gálvez, Juan; de la Torre, Javier; Hidalgo-Tenorio, Carmen; Reguera, José M; Márquez, Manuel; Martínez-Marcos, Francisco; de Alarcón, Arístides

2008-05-01

To describe the epidemiological, clinical, and prognostic characteristics of patients with left-sided native valve endocarditis (LNVE) caused by coagulase-negative staphylococci (CoNS). Prospective multicenter study of endocarditis cases reported in the Andalusian Cohort for the Study of Cardiovascular Infections between 1984 and 2005. Among 470 cases of LNVE, 39 (8.3%) were caused by CoNS, a number indicating a 30% increase in the incidence of this infection over the last decade. The mean age of affected patients was 58.32 +/- 15 years and 27 (69.2%) were men. Twenty-one patients (53.8%) had previous known valve disease and half the episodes were considered nosocomial (90% of them from vascular procedures). Median time interval from the onset of symptoms to diagnosis was 14 days (range: 1-120). Renal failure (21 cases, 53.8%), intracardiac damage (11 cases, 28.2%), and central nervous system involvement (10 cases, 25.6%) were the most frequent complications. There were only 3 cases (7.7%) of septic shock. Surgery was performed in 18 patients (46.2%). Nine patients (23.1%) died, overall. Factors associated with higher mortality in the univariate analysis were acute renal failure (P = 0.023), left-sided ventricular failure (P = 0.047), and time prior to diagnosis less than 21 days (P = 0.018). As compared to LNVE due to other microorganisms, the patients were older (P = 0.018), had experienced previous nosocomial manipulation as the source of bacteremia (P < 0.001), and developed acute renal failure more frequently (P = 0.001). Mortality of LNVE due to CoNS was lower than mortality in Staphylococcus aureus infection, but higher than in Streptococcus viridans infection. Left-sided native valve endocarditis due to CoNS is now increasing because of the ageing of the population. This implies more frequent invasive procedures (mainly vascular) as a consequence of the concomitant disease. Nonetheless, the mortality associated with LNVE due to CoNS does not seem to be

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

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

Factors associated with methicillin-resistant coagulase-negative staphylococci as causing organisms in deep sternal wound infections after cardiac surgery

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R. Sommerstein

2015-07-01

Full Text Available Established preoperative antibiotic prophylaxis in cardiac surgery is ineffective against methicillin-resistant coagulase-negative staphylococci (CoNS. This case–control study aimed to determine factors predicting deep sternal wound infections due to methicillin-resistant CoNS. All cardiac surgery patients undergoing sternotomy between June 2009 and March 2013 prospectively documented in a Swiss tertiary care center were included. Among 1999 patients, 82 (4.1% developed deep sternal wound infection. CoNS were causal in 36 (44% patients, with 25/36 (69% being methicillin resistant. Early reintervention for noninfectious causes (odds ratio (OR 4.3; 95% confidence interval (CI 1.9–9.5 was associated with methicillin-resistant CoNS deep sternal wound infection. Among CoNS deep sternal wound infection, perioperative antimicrobial therapy (p 0.002, early reintervention for noninfectious causes (OR 7.9; 95% CI 0.9–71.1 and time between surgery and diagnosis of infection over 21 days (OR 10.8; 95% CI 1.2–97.8 were associated with methicillin resistance. These findings may help to better tailor preoperative antimicrobial prophylaxis.

Frequency of antiseptic resistance genes in clinical staphycocci and enterococci isolates in Turkey

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Seyda Ignak

2017-08-01

Full Text Available Abstract Background Disinfectants and antiseptics are biocides widely used in hospitals to prevent spread of pathogens. It has been reported that antiseptic resistance genes, qac’s, caused tolerance to a variety of biocidal agents, such as benzalkonium chloride (BAC and chlorhexidine digluconate (CHDG in Staphylococcus spp. isolates. We aimed to search the frequency of antiseptic resistance genes in clinical Staphylococcus spp. and Enterococcus spp. isolates to investigate the possible association with antiseptic tolerance and antibiotic resistance. Methods Antiseptic resistance genes (qacA/B, smr, qacG, qacH, and qacJ isolated from Gram-positive cocci (69 Staphylococcus spp. and 69 Enterococcus spp. were analyzed by PCR method. The minimum inhibitory concentrations (MICs of BAC and CHDG were determined by agar dilution method, whereas antibiotic susceptibility was analyzed by disk diffusion method according to Clinical and Laboratory Standards Institute (CLSI criteria. Results The frequency of antiseptic resistance genes was found to be high (49/69; 71.0% in our clinical staphylococci isolates but absent (0/69; 0% in enterococci isolates. The frequency of qacA/B and smr genes was higher (25/40; 62.5% and 7/40; 17.5%, respectively in coagulase negative staphylococci (CNS when compared to Staphylococcus aureus strains (3/29; 10.3%, and 4/29; 13.8%, respectively. In contrast, the frequency of qacG and qacJ genes was higher (11/29; 37.9% and 8/29; 27.5%, respectively in S. aureus than those of CNS (5/40; 12.5%, 10/40; 25.0% strains. qacH was not identified in none of the strains. We found an association between presence of antiseptic resistance genes and increased MIC values of BAC (>4 μg/mL in staphylococci and it was found to be statistically statistically significant (p < 0.01. We also showed that MICs of BAC and CHDG of vancomycin-resistant enterococci (VRE isolates were significantly higher than those of vancomycin

Trends and molecular mechanisms of antimicrobial resistance in clinical staphylococci isolated from companion animals over a 16 year period.

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Couto, Natacha; Monchique, Cláudia; Belas, Adriana; Marques, Cátia; Gama, Luís T; Pomba, Constança

2016-06-01

The objective of this study was to investigate the evolution of resistance to antimicrobials, corresponding mechanisms and molecular characteristics of Staphylococcus spp., between 1999 and 2014. Susceptibility to 38 antimicrobials was determined for 632 clinical staphylococcal isolates obtained from companion animals (dogs, cats, horses and other animals). Twenty antimicrobial resistance genes, including mecA and mecC, were screened by PCR. Methicillin-resistant staphylococci were characterized by spa (Staphylococcus aureus), SCCmec, MLST and PFGE typing. Statistical analyses were performed using SAS v9.3 and differences were considered relevant if P ≤ 0.05. The mecA gene was identified in 74 staphylococcal isolates (11.6%): 11 MRSA (40.7%), 40 methicillin-resistant Staphylococcus pseudintermedius (MRSP; 8.7%) and 23 methicillin-resistant CoNS (26.7%). Resistance to the majority of antimicrobials and the number of mecA-positive isolates increased significantly over time. Eighteen spa types were identified, including two new ones. MRSA isolates were divided into three PFGE clusters that included ST22-IV, ST105-II, ST398-V and ST5-VI. Most methicillin-resistant Staphylococcus epidermidis isolates were of clonal complex (CC) 5, including a new ST, and clustered in eight PFGE clusters. MRSP were grouped into five PFGE clusters and included ST45-NT, ST71-II-III, ST195-III, ST196-V, ST339-NT, ST342-IV and the new ST400-III. Methicillin-resistant Staphylococcus haemolyticus clustered in two PFGE clusters. The significant increase in antimicrobial-resistant and mecA-positive isolates in recent years is worrying. Furthermore, several isolates are MDR, which complicates antimicrobial treatment and increases the risk of transfer to humans or human isolates. Several clonal lineages of MRSA and methicillin-resistant S. epidermidis circulating in human hospitals and the community were found, suggesting that companion animals can become infected with and contribute to the

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

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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

Cultural characters of a newly recognized group of hospital staphylococci.

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Jevons, M P; John, M; Parker, M T

1966-07-01

Members of a newly recognized group of hospital staphylococci, which are believed to have arisen from 83A staphylococci by lysogenization, differ from them in several cultural characters. Some but not all of these characters appear to be determined by the carriage of phage.

Short communication: Species group-specific predictors at the cow and quarter level for intramammary infection with coagulase-negative staphylococci in dairy cattle throughout lactation.

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De Visscher, A; Piepers, S; Supré, K; Haesebrouck, F; De Vliegher, S

2015-08-01

Coagulase-negative staphylococci (CNS) are frequently isolated from quarters with subclinical mastitis, teat apices, and the cows' environment. Virulence, ecology, epidemiological behavior, and effect on udder health vary between different CNS species. Staphylococcus chromogenes, Staph. simulans, and Staph. xylosus are frequently present in milk and have a more substantial effect on quarter milk somatic cell count than other species. Therefore, these species are considered the "more relevant" CNS. As species-specific factors associated with CNS intramammary infection (IMI) have not yet been identified and susceptibility for IMI differs between cows and quarters, this study aimed to identify predictors for CNS IMI at the cow and quarter level (some of them changing over time) with a specific focus on the aforementioned more relevant CNS. Precise data were available from a longitudinal study (3,052 observations from 344 quarters from 86 dairy cows belonging to 3 commercial dairy herds). All CNS were molecularly identified to the species level, and multivariable, multilevel logistic regression models taking into account the longitudinal nature of the data, were fit to study the likelihood of infection. Staphylococcus chromogenes, Staph. xylosus, and Staph. cohnii were the most frequently isolated species from CNS IMI in older cows, whereas Staph. chromogenes, Staph. xylosus, and Staph. simulans were the main species found in IMI in heifers. Quarters from heifers (as opposed to multiparous cows), from heifers and multiparous cows in third or fourth month in lactation (as opposed to early lactation, <60 d in milk), and with an increasing quarter milk SCC were more likely to be infected with the more relevant CNS species. Quarter milk SCC was identified as the sole statistically significant predictor for IMI with other CNS species, although the size of the effect was lower [odds ratio of 1.6 (1.4-1.9) vs. 2.1 (1.8-2.5)] than the effect for IMI with the more relevant CNS

Identification of a novel vga(E) gene variant that confers resistance to pleuromutilins, lincosamides and streptogramin A antibiotics in staphylococci of porcine origin.

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Li, Jun; Li, Beibei; Wendlandt, Sarah; Schwarz, Stefan; Wang, Yang; Wu, Congming; Ma, Zhiyong; Shen, Jianzhong

2014-04-01

To investigate the genetic basis of pleuromutilin resistance in coagulase-negative staphylococci of porcine origin that do not carry known pleuromutilin resistance genes and to determine the localization and genetic environment of the identified resistance gene. Plasmid DNA of two pleuromutilin-resistant Staphylococcus cohnii and Staphylococcus simulans isolates was transformed into Staphylococcus aureus RN4220. The identified resistance plasmids were sequenced completely. The candidate gene for pleuromutilin resistance was cloned into shuttle vector pAM401. S. aureus RN4220 transformants carrying this recombinant shuttle vector were tested for their MICs. S. cohnii isolate SA-7 and S. simulans isolate SSI1 carried the same plasmid of 5584 bp, designated pSA-7. A variant of the vga(E) gene was detected, which encodes a 524 amino acid ATP-binding cassette protein. The variant gene shared 85.7% nucleotide sequence identity and the variant protein 85.3% amino acid sequence identity with the original vga(E) gene and Vga(E) protein, respectively. The Vga(E) variant conferred cross-resistance to pleuromutilins, lincosamides and streptogramin A antibiotics. Plasmid pSA-7 showed an organization similar to that of the apmA-carrying plasmid pKKS49 from methicillin-resistant S. aureus and the dfrK-carrying plasmid pKKS966 from Staphylococcus hyicus. Sequence comparisons suggested that recombination events may have played a role in the acquisition of this vga(E) variant. A novel vga(E) gene variant was identified, which was located on a small plasmid and was not associated with the transposon Tn6133 [in contrast to the original vga(E) gene]. The plasmid location may enable its further dissemination to other staphylococci and possibly also to other bacteria.

Antibacterial susceptibility profiles of subclinical mastitis pathogens isolated in Batna and Setif Governorates (East of Algeria

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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

SYNERGISTIC EFFECTS OF ETHANOL MEDICINAL PLANT EXTRACTS WITH ERYTHROMYCIN AGAINST SKIN STRAINS OF STAPHYLOCOCCI WITH INDUCIBLE PHENOTYPE OF MLS-RESISTANCE

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Yurchyshyn O.I.

2017-10-01

decreased in 32-64 times and Geranium pratense L. rhizomes extract antimicrobial concentration was decreased in 64-256 times. Ethanol extracts of Betula verrucosa L. buds (average FICI 0.473±0.20, Arctostaphylos uva-ursi (L. Spreng. leaves (average FICI 0.143±0.18 and Tamarix ramosissima Ledeb. leaves (average FICI 0.189±0.29 showed synergic action with erythromycin against 71.4-85.7% tested strains. Ethanol extracts of Sanguisorba officinalis L. roots showed non-interactive action with antibiotic against 42.8% isolates of staphylococci. Additive interaction with erythromycin for this extract was observed against 28.6% and synergic action against 28.6% strains (average FICI 0.812±0.52. Biota orientalis (L. Endl. (Platycladus orientalis (L. Franco fruits extract and Cotinus coggygria Scop. (Rhus cotinus R. leaves extract exhibited non-interactive action with antibiotic against all tested strains (average FICI 2.0±0.0. Experimental data indicate that combination of plant extracts with macrolides in therapeutic regimens against MLS-resistant staphylococci is promising, particularly for the treatment of pyoderma. The introduction of combined chemotherapy in clinical practice can actually help to solve two problems of modern medicine - slow the process of microorganisms (such as staphylococcus resistance to antibiotics acquiring and improve treatment of infections caused by resistant strains. Detection of bacteria antibiotic resistance modifiers in various plants stimulates to their intensive phytochemical study for isolation and identification of the active components. It will help to investigate the mechanisms of synergy on the molecular level. Conclusion. BAC of medicinal and aromatic plants potentiate antimicrobial activity of macrolides against skin isolates of staphylococci with inducible MLS-resistance. Alnus incana L. fruits ethanolic extract demonstrates the best direct antimicrobial activity and in combination with ERY synergistically inhibits the growth of S

The diversity of antimicrobial resistance genes among staphylococci of animal origin.

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Wendlandt, Sarah; Feßler, Andrea T; Monecke, Stefan; Ehricht, Ralf; Schwarz, Stefan; Kadlec, Kristina

2013-08-01

Staphylococci of animal origin harbor a wide variety of resistance genes. So far, more than 40 different resistance genes have been identified in staphylococci from animals. This includes genes that confer resistance to virtually all classes of antimicrobial agents approved for use in animals, such as penicillins, cephalosporins, tetracyclines, macrolides, lincosamides, phenicols, aminoglycosides, aminocyclitols, pleuromutilins, and diaminopyrimidines. The gene products of some of these resistance genes confer resistance to only specific members of a class of antimicrobial agents, whereas others confer resistance to the entire class or even to members of different classes of antimicrobial agents. The resistance mechanisms specified by the resistance genes fall into three major categories: (i) enzymatic inactivation, (ii) active efflux, or (iii) protection/modification/replacement of the cellular target sites of the antimicrobial agents. Mobile genetic elements, in particular plasmids and transposons, play a major role as carriers of antimicrobial resistance genes in animal staphylococci. They facilitate the exchange of resistance genes with staphylococci of human origin but also with other Gram-positive bacteria. Copyright © 2013 Elsevier GmbH. All rights reserved.

Identification of Clinical Staphylococcal Isolates from Humans by Internal Transcribed Spacer PCR

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Couto, Isabel; Pereira, Sandro; Miragaia, Maria; Sanches, Ilda Santos; de Lencastre, Hermínia

2001-01-01

The emergence of coagulase-negative staphylococci not only as human pathogens but also as reservoirs of antibiotic resistance determinants requires the deployment and development of methods for their rapid and reliable identification. Internal transcribed spacer-PCR (ITS-PCR) was used to identify a collection of 617 clinical staphylococcal isolates. The amplicons were resolved in high-resolution agarose gels and visually compared with the patterns obtained for the control strains of 29 staphylococcal species. Of the 617 isolates studied, 592 (95.95%) were identified by ITS-PCR and included 11 species: 302 isolates of Staphylococcus epidermidis, 157 of S. haemolyticus, 79 of S. aureus, 21 of S. hominis, 14 of S. saprophyticus, 8 of S. warneri, 6 of S. simulans, 2 of S. lugdunensis, and 1 each of S. caprae, S. carnosus, and S. cohnii. All species analyzed had unique ITS-PCR patterns, although some were very similar, namely, the group S. saprophyticus, S. cohnii, S. gallinarum, S. xylosus, S. lentus, S. equorum, and S. chromogenes, the pair S. schleiferi and S. vitulus, and the pair S. piscifermentans and S. carnosus. Four species, S. aureus, S. caprae, S. haemolyticus, and S. lugdunensis, showed polymorphisms on their ITS-PCR patterns. ITS-PCR proved to be a valuable alternative for the identification of staphylococci, offering, within the same response time and at lower cost, higher reliability than the currently available commercial systems. PMID:11526135

Identification of clinical staphylococcal isolates from humans by internal transcribed spacer PCR.

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Couto, I; Pereira, S; Miragaia, M; Sanches, I S; de Lencastre, H

2001-09-01

The emergence of coagulase-negative staphylococci not only as human pathogens but also as reservoirs of antibiotic resistance determinants requires the deployment and development of methods for their rapid and reliable identification. Internal transcribed spacer-PCR (ITS-PCR) was used to identify a collection of 617 clinical staphylococcal isolates. The amplicons were resolved in high-resolution agarose gels and visually compared with the patterns obtained for the control strains of 29 staphylococcal species. Of the 617 isolates studied, 592 (95.95%) were identified by ITS-PCR and included 11 species: 302 isolates of Staphylococcus epidermidis, 157 of S. haemolyticus, 79 of S. aureus, 21 of S. hominis, 14 of S. saprophyticus, 8 of S. warneri, 6 of S. simulans, 2 of S. lugdunensis, and 1 each of S. caprae, S. carnosus, and S. cohnii. All species analyzed had unique ITS-PCR patterns, although some were very similar, namely, the group S. saprophyticus, S. cohnii, S. gallinarum, S. xylosus, S. lentus, S. equorum, and S. chromogenes, the pair S. schleiferi and S. vitulus, and the pair S. piscifermentans and S. carnosus. Four species, S. aureus, S. caprae, S. haemolyticus, and S. lugdunensis, showed polymorphisms on their ITS-PCR patterns. ITS-PCR proved to be a valuable alternative for the identification of staphylococci, offering, within the same response time and at lower cost, higher reliability than the currently available commercial systems.

Further studies on staphylococci in meats. III. Occurrence and characteristics of coagulase-positive strains from a variety of nonfrozen market cuts.

Science.gov (United States)

JAY, J M

1962-05-01

From 34 retail grocery stores and meat markets, 209 samples of nonfrozen meats were obtained and analyzed for coagulase-positive Staphylococcus aureus, employing six selective media. Sixty-seven (38.7%) of 173 samples obtained from 27 stores yielded S. aureus. No coagulase-positive S. aureus was isolated from 36 samples obtained from 7 of the stores. The 67 meats yielded 272 isolates from 10 different kinds of meats. There were 162 physiological strains represented when classified by store and 36 strains classified without regard to store of origin. The larger stores yielded fewer meats with staphylococci than the smaller stores. The meats from which S. aureus was recovered in the order of frequency of percentage recovery are as follows: chicken, pork liver, fish, spiced ham, round beef steak, hamburger, beef liver, pork chops, veal steak, and lamb chops. The following seven meats did not yield staphylococci: bologna, shucked oysters, olive and pickle loaf, salami, wieners, and chopped ham. Eighty-eight per cent of the isolates produced pigment, 85% were gelatinase positive, only 1 strain failed to form a precipitate on egg yolk agar, 92% formed deoxyribonuclease, 87% produced bound coagulase, 91% produced the alpha-hemolysin, 70% the delta-, 22% the beta-, and 6% were nil in this regard. The isolates are compared with hospital and other food strains, and their possible source in the meats is discussed.

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...

Prevalence of the antibiotic resistance genes in coagulase-positive- and negative-Staphylococcus in chicken meat retailed to consumers

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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

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, mec A 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, mec A 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 mec A 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

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.

Antibiotic resistance of staphylococci from humans, food and different animal species according to data of the Hungarian resistance monitoring system in 2001.

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Kaszanyitzky, Eva J; Jánosi, Sz; Egyed, Zsuzsanna; Agost, Gizella; Semjén, G

2003-01-01

Based on data of the Hungarian resistance monitoring system the antibiotic resistance of Staphylococcus strains of human and animal origin was studied. No methicillin-resistant staphylococci harbouring mecA gene were isolated from animals in 2001. Penicillin resistance, mediated by penicillinase production, was the most frequent among Staphylococcus aureus strains isolated from humans (96%), from bovine mastitis (55%), from foods (45%) and from dogs. In staphylococci isolated from animals low resistance percentages to aminoglycosides (0-2%), fluoroquinolones (0.5-3%) and sulphonamides (0.5-4%) were found but in strains isolated humans these figures were higher (1-14%, 5-18% and 3-31%, respectively). The most frequent antibiotic resistance profiles of strains isolated from animals and food were penicillin/tetracycline, penicillin/lincomycin and penicillin/lincomycin/tetracycline. Penicillin/tetracycline resistance was exhibited by strains from mastitis (3), samples from the meat industry (31), poultry flocks (1), poultry industry (1), noodle (1) and horses (2). Penicillin/lincomycin resistance was found in 10 Staphylococcus strains from mastitis, 1 from the dairy industry, 1 from the meat industry and 6 from dogs. Isolates from mastitis (2), from the dairy industry (2), from pigs (1), from the meat industry (1) and from poultry (1) harboured penicillin/lincomycin/tetracycline resistance pattern. Multiresistant strains were usually isolated only from one and sometimes from two animal species; therefore, the spread of defined resistant strains (clones) among different animal species could not be demonstrated. These results also suggest that the transfer of antibiotic resistance of S. aureus from animals to humans probably occurs less frequently than is generally assumed.

Comparison phenotypic and genotypic identification of Staphylococcus species isolated from bovine mastitis

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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

Isolation and molecular characterization of a urease-negative Actinobacillus pleuropneumoniae mutant.

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Ito, Hiroya; Takahashi, Sayaka; Asai, Tetsuo; Tamura, Yutaka; Yamamoto, Koshi

2018-01-01

An atypical urease-negative mutant of Actinobacillus pleuropneumoniae serovar 2 was isolated in Japan. Nucleotide sequence analysis of the urease gene cluster revealed that the insertion of a short DNA sequence into the cbiM gene was responsible for the urease-negative activity of the mutant. Veterinary diagnostic laboratories should be watchful for the presence of aberrant urease-negative A. pleuropneumoniae isolates.

The prevalence of carriage of meticillin-resistant staphylococci by veterinary dermatology practice staff and their respective pets.

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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. © 2010 The Authors. Journal compilation © 2010 ESVD and ACVD.

Reliability of the MicroScan WalkAway PC21 panel in identifying and detecting oxacillin resistance in clinical coagulase-negative staphylococci strains.

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Olendzki, A N; Barros, E M; Laport, M S; Dos Santos, K R N; Giambiagi-Demarval, M

2014-01-01

The purpose of this study was to determine the reliability of the MicroScan WalkAway PosCombo21 (PC21) system for the identification of coagulase-negative staphylococci (CNS) strains and the detection of oxacillin resistance. Using molecular and phenotypic methods, 196 clinical strains were evaluated. The automated system demonstrated 100 % reliability for the identification of the clinical strains Staphylococcus haemolyticus, Staphylococcus hominis and Staphylococcus cohnii; 98.03 % reliability for the identification of Staphylococcus epidermidis; 70 % reliability for the identification of Staphylococcus lugdunensis; 40 % reliability for the identification of Staphylococcus warneri; and 28.57 % reliability for the identification of Staphylococcus capitis, but no reliability for the identification of Staphylococcus auricularis, Staphylococcus simulans and Staphylococcus xylosus. We concluded that the automated system provides accurate results for the more common CNS species but often fails to accurately identify less prevalent species. For the detection of oxacillin resistance, the automated system showed 100 % specificity and 90.22 % sensitivity. Thus, the PC21 panel detects oxacillin-resistant strains, but is limited by the heteroresistance that is observed when using most phenotypic methods.

Clinical significance of the isolation of Staphylococcus epidermidis from bone biopsy in diabetic foot osteomyelitis

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Javier Aragón-Sánchez

2010-08-01

Full Text Available Introduction: Coagulase-negative staphylococci are considered as microorganisms with little virulence and usually as contaminants. In order to establish the role of Staphylococcus epidermidis as a pathogen in diabetic foot osteomyelitis, in addition to the isolation of the sole bacterium from the bone it will be necessary to demonstrate the histopathological changes caused by the infection. Methods: A consecutive series of 222 diabetic patients with foot osteomyelitis treated surgically in the Diabetic Foot Unit at La Paloma Hospital (Las Palmas de Gran Canaria, Canary Islands, Spain between 1 October 2002 and 31 October 2008. From the entire series including 213 bone cultures with 241 isolated organisms, we have analyzed only the 139 cases where Staphylococci were found. We analyzed several variables between the two groups: Staphylococcus aureus versus Staphylococcus epidermidis. Results: Of the 134 patients included in this study, Staphlylococcus epidermidis was found as the sole bacterium isolated in 11 cases and accompanied by other bacteria in 12 cases. Staphlylococcus aureus was found as the sole bacterium isolated in 72 cases and accompanied by other bacteria in 39 cases. Histopathological changes were found in the cases of osteomyelitis where Staphylococcus epidermidis was the sole bacterium isolated. Acute osteomyelitis was found to a lesser extent when Staphylococcus epidermidis was the sole bacterium isolated but without significant differences with the cases where Staphylococcus aureus was the sole bacterium isolated. Conclusion: Staphylococcus epidermidis should be considered as a real pathogen, not only a contaminant, in diabetic patients with foot osteomyelitis when the bacterium is isolated from the bone. No differences in the outcomes of surgical treatment have been found with cases which Staphlylococcus aureus was isolated.

Virulence factors genes of Staphylococcus spp. isolated from caprine subclinical mastitis.

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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.

Unraveling the microbiota of teat apices of clinically healthy lactating dairy cows, with special emphasis on coagulase-negative staphylococci.

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Braem, G; De Vliegher, S; Verbist, B; Piessens, V; Van Coillie, E; De Vuyst, L; Leroy, F

2013-03-01

Swab samples (n=72) obtained from the teat apex of lactating dairy cows without visual signs of inflammation (n=18) were gathered on 2 well-managed Flemish dairy herds (herds 1 and 2) during the same month to assess the bacterial diversity of teat apices before milking. A combination of both culture-dependent [plating and (GTG)(5)-PCR fingerprinting of the colonies] and culture-independent [denaturing gradient gel electrophoresis (PCR-DGGE)] techniques indicated that the teat apices contain a wide diversity of bacterial genera. Despite a low bacterial load, 20 bacterial genera of 3 phyla (Actinobacteria, Firmicutes, and Proteobacteria) were present. The most prevalent bacteria were the coagulase-negative staphylococci (CNS), encompassing a total of 15 species, which were identified to the species level using a combination of (GTG)(5)-PCR fingerprinting, gene sequencing (16S ribosomal RNA and rpoB genes), and a novel PCR-DGGE technique based on the tuf-PCR amplicon. Overall bacterial diversity did not differ significantly between the herds or between noninfected and subclinically infected quarters in herd 1. In herd 1, borderline significant lower CNS species diversity was found on teat apices of noninfected quarters compared with subclinically infected quarters. The most prevalent CNS species were Staphylococcus haemolyticus and Staphylococcus equorum in both herds and Staphylococcus carnosus in herd 2. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Antibacterial activities of coagulase-negative staphylococci from bovine teat apex skin and their inhibitory effect on mastitis-related pathogens.

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Braem, G; Stijlemans, B; Van Haken, W; De Vliegher, S; De Vuyst, L; Leroy, F

2014-05-01

To explore antibacterial activities of coagulase-negative staphylococci (CoNS) from teat apices of dairy cows towards mastitis-causing pathogens. Of 254 CoNS, 38 displayed bacteriocin-like activity after a first screening. Seven of these strains displayed activity against at least one mastitis-related pathogen (Streptococcus uberis, Streptococcus dysgalactiae and Staphylococcus aureus). Staphylococcus chromogenes L217 displayed the strongest inhibitory effect, being active against all tested mastitis-related pathogens and most tested CoNS. Based on cation exchange and reversed-phase chromatography, in addition to N-terminal Edman degradation and PCR, the antibacterial peptide was identified as a nukacin-type bacteriocin and named nukacin L217. Although staphylococcal bacteriocins are generally found in the cell-free supernatants of liquid cultures, Staph. chromogenes L217 only led to detectable activity when grown on agar medium. Bacteriocin-like activities are not uncommon among CoNS from teat apices and may inhibit mastitis-causing pathogens, as found for nukacin L217 production by Staph. chromogenes L217. Nukacin L217 is the first identified bacteriocin of the species Staph. chromogenes and displays unusual production kinetics, that is, requiring surface growth of its producer. The fact that nukacins are produced by different CoNS species suggests a role in the teat skin ecosystem. © 2014 The Society for Applied Microbiology.

Occurrence and characterization of Staphylococcus bacteria isolated from poultry in Western Poland.

Science.gov (United States)

Marek, Agnieszka; Stepień-Pyśniak, Dagmara; Pyzik, Ewelina; Adaszek, Łukasz; Wilczyński, Jarosław; Winiarczyk, Stanisław

2016-01-01

In the pathology of poultry, infections caused by Staphylococcus spp. are taking on increasing significance. Although the Staphylococcus species most frequently isolated from these animals is Staphylococcus aureus, the literature data indicate that other species, both coagulase-positive and coagulase-negative, can also cause infections in birds. The aim of the study was to assess the frequency of occurrence of Staphylococcus infections in various poultry species in Western Poland and to test the susceptibility of isolated strains to selected antibiotics. The results obtained showed a relatively high rate of Staphylococcus infection in the poultry. From 2805 samples tested 302 strains (10.8%) of Staphylococcus were isolated. As many as 25 Staphylococcus species were distinguished among the strains isolated. S. cohnii (23.50%), S. aureus (15.89%) and S. lentus (13.90%) accounted for the highest percentages. Over half of the isolated staphylococci exhibited resistance to five of the antibiotics applied, with the highest percentage of resistant strains, 65%, noted for enrofloxacin.

Tigecycline activity tested against 26,474 bloodstream infection isolates: a collection from 6 continents.

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Sader, Helio S; Jones, Ronald N; Stilwell, Matthew G; Dowzicky, Michael J; Fritsche, Thomas R

2005-07-01

The activity of tigecycline (formerly GAR936), a novel glycylcycline, was tested against recent bloodstream infection (BSI) pathogen isolates from 6 continents. Frequency of clinical occurrence of these pathogens was determined and their antibiograms assessed using reference broth microdilution methods. A total of 26474 strains were tested for tigecycline susceptibility according to the Clinical and Laboratory Standards Institute (formerly the National Committee for Clinical Laboratory Standards) by the M7-A6 guidelines with interpretations from M100-S15 and the package insert. The rank order of pathogens was Staphylococcus aureus (33.1%), Escherichia coli (14.0%), coagulase-negative staphylococci (13.5%), Enterococcus spp. (12.3%), Klebsiella spp. (5.7%), Pseudomonas aeruginosa (4.2%), Enterobacter spp. (3.0%), beta-hemolytic streptococci (2.9%), Streptococcus pneumoniae (2.3%), and viridans group streptococci (1.4%). Tigecycline exhibited a broader spectrum of activity against BSI isolates when compared to ciprofloxacin, tetracycline, aminoglycosides, and many beta-lactams (imipenem). Tigecycline was highly active against most pathogens tested, including staphylococci (MIC(90), 0.5 microg/mL), enterococci (MIC90, 0.25 microg/mL), streptococci (MIC(90), < or =0.12 microg/mL), Escherichia coli (MIC90, 0.25 microg/mL), Klebsiella spp. (MIC90, 1 mmicrog/mL), and Enterobacter spp. (MIC(90), 2 mmicrog/mL), but showed limited inhibition of Pseudomonas aeruginosa (MIC90, 16 microg/mL) and indole-positive or indole-negative Proteae (MIC90, 4-8 microg/mL). In summary, tigecycline exhibited a wide spectrum of antimicrobial potency versus BSI isolates collected worldwide. Serious infections in nosocomial environments should benefit from tigecycline use among the investigational phase 3 agents focused toward resistant strains.

A study of bacterial isolates from corneal specimens and their antibiotic resistance profile

International Nuclear Information System (INIS)

Begum, N. N.; Al-Khattaf, Abdulaziz S.; Yeboah, E. A.; Kambal, Abdel-Majed M.; Al-Mansouri, Samir M.

2006-01-01

We aim to examine the spectrum of bacteria causing corneal infections and their antibiotic susceptibility patterns. This will serve as a guideline for empiric therapy of corneal infections. We conducted the study over a period of 18 months from March 2001 through December 2002 in King Abdul-Aziz University Hospital, Riyadh, Kingdom of Saudi Arabia. Corneal specimens taken from 200 patients were inoculated directly onto different types of media. The isolates were identified and then tested against the appropriate topical or systemic antibiotics. Sixty-seven (33.5%) of the total specimens were culture positive and 133 (66.5%) were culture negative. Fourteen (7%) of these showed organisms in the Gram stained smears and correlated well with the culture reports. Of the 67 positive cultures, 53 (79.1%) were Gram-positive bacteria mostly coagulase-negative Staphylococci 29 (43.3%) followed by Streptococcus pneumoniae (S. pneumoniae) 13 (19.4%). Among Gram-negative bacteria 14 (20.9%), Pseudomonas aeruginosa (P. aeruginosa) 10 (14.9%) was the predominant isolate. All the isolates were sensitive to ofloxacin and the commonly used ocular antibiotics. All the isolated bacteria were sensitive to ofloxacin, a fluoroquinolone. Having marked potency for broad-spectrum activity against both Gram-positive and Gram-negative bacteria, make the fluoroquinolones especially the newer generations, a potential single drug therapy for corneal infections. (author)

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

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Basso, Ana P; Martins, Paula D; Nachtigall, Gisele; Van Der Sand, Sueli; De Moura, Tiane M; Frazzon, Ana Paula G

2014-12-01

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.

Characterization of Staphylococcus spp. strains in milk from buffaloes with mastitis in Brazil: the need to identify to species level to avoid misidentification

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V. Coimbra-e-Souza

Full Text Available ABSTRACT Mastitis is an inflammation of the mammary gland that affects dairy cattle worldwide causing economic losses. Coagulase-negative staphylococci (CNS are the predominant cause of this type of infection. We have recently showed that coagulase-positive staphylococci could be misidentified. So, the aim of this study was to characterize the Staphylococcus spp. strains initially classified as coagulase-negative Staphylococci, isolated from buffalo with subclinical mastitis. Milk of buffaloes with mastitis in herds was collected and 9 strains were identified as CNS by phenotypic tests. Molecular methodologies latter identified the strains as coagulase-negative Staphylococcus chromogenes (5, coagulase-positive Staphylococcus hyicus (2 and coagulase-positive Staphylococcus aureus (2. Our results strongly support the need to identify the isolates to a species level in order to avoid misidentification and to be aware of the classification using the coagulase test alone.

Antimicrobial susceptibility pattern and plasmid-mediated ...

African Journals Online (AJOL)

negative Staphylococci (CoNS) were isolated from clinical samples and isolates subjected to antibiotic susceptibility testing, plasmid curing and plasmid DNA isolation. Result: The highest percentages isolates were recovered from urine samples and ...

Negative masses, even if isolated, imply self-acceleration, hence a catastrophic world

International Nuclear Information System (INIS)

Cavalleri, G.; Tonni, E.

1997-01-01

The conjecture of the existence of negative masses together with ordinary positive masses leads to runaway motions even if no self-reaction is considered. Pollard and Dunning-Davies have shown other constraints as a modification of the principle of least action and that negative masses can only exist at negative temperature, and must be adiabatically separate from positive masses. They show here that the self-reaction on a single isolated negative mass implies a runaway motion. Consequently, the consideration of self-fields and relevant self-reaction excludes negative masses even if isolated

Prevalence of antiseptic resistance genes increases in staphylococcal isolates from orthokeratology lens wearers over initial six-month period of use.

Science.gov (United States)

Guang-Sen, Shi; Boost, Maureen; Cho, Pauline

2016-06-01

The purpose of this study was to investigate the prevalence of antiseptic-resistance (QAC) genes in staphylococci colonizing periorbital tissues and accessories of orthokeratology (ortho-k) lens wearers over a 6-month period and determine the effect of their presence on minimum inhibitory (MIC) and bactericidal concentrations (MBC) of disinfectants and log reduction of multipurpose contact lens solutions (MPS). Staphylococci were isolated from periorbital tissues and accessories of patients before commencing ortho-k therapy and at two subsequent visits. Presence of QAC genes in 116 S. aureus and 67 CNS isolates was determined by PCR and association with period of ortho-k use determined. MICs and MBCs of staphylococci gene-positive were compared with gene-negative strains and the effectiveness of four MPS for rigid contact lenses investigated. S. aureus carriage rates in the conjunctiva decreased significantly from 41.2 % (baseline) to 11.8 % (3-months), and 13.3 % (6-months) (p trend 0.03), while CNS increased from 58.8 %(baseline) to 94.1 % (3-months), and 93.3 % (6-months) (p trend 0.02). Prevalence of qacA/B increased considerably over time (S. aureus: 4.4 % to 15.4 %, CNS: 6.7 % to 25 %), but frequency of smr was relatively stable. Only five CNS isolates harboured qacH. MICs and MBCs of gene-positive isolates were significantly increased and three MPS did not achieve a 3-log reduction of many QAC-positive strains. Ortho-k lens wear contributed to changes of staphylococcal carriage rates in the conjunctival sac. Use of MPS containing quaternary ammonium compounds may select for the carriage of organisms harbouring QAC genes, as the low concentration of disinfecting agents is not adequate for killing gene-positive strains.

Gevoeligheid voor diverse antibiotica van coagulase negatieve staphylococcen, geisoleerd uit melkmonsters van Nederlands rundvee

NARCIS (Netherlands)

Sampimon, O.C.; Vernooij, J.C.A.; Mevius, D.J.; Sol, J.

2007-01-01

During recent years the prevalence of coagulase-negative staphylococci in milk samples from Dutch dairy cows has increased. In 1999 16.2% of the bacteria isolated from milk collected from cows with subclinical mastitis were coagulasenegative staphylococci. In 2004 this proportion was 42.2%. The

Further evidence for the existence of environmental and host-associated species of coagulase-negative staphylococci in dairy cattle.

Science.gov (United States)

De Visscher, Anneleen; Supré, Karlien; Haesebrouck, Freddy; Zadoks, Ruth N; Piessens, Veerle; Van Coillie, Els; Piepers, Sofie; De Vliegher, Sarne

2014-08-27

Coagulase-negative staphylococci (CNS) are abundantly present in the dairy farm environment and on bovine skin and mucosae. They are also the most prevalent bacteria causing bovine intramammary infections (IMI). Reservoirs and transmission routes of CNS are not yet fully unraveled. The objectives of this study were to explore the distribution of CNS in parlor-related extramammary niches and to compare it to the distributions of CNS causing IMI in those herds. Niches that were targeted in this study were cows' teat apices, milking machine unit liners, and milker's skin or gloves. Each of the three herds had its own CNS microbiota in those niches. The most prevalent species in the parlor-related extramammary niches were Staphylococcus cohnii, S. fleurettii, and S. equorum in the first, second, and third herd, respectively, whereas S. haemolyticus and S. sciuri were found in all herds. S. cohnii and S. fleurettii, as well as S. haemolyticus, which was present in each herd, were also frequently found in milk samples. By contrast, S. chromogenes, S. simulans, and S. xylosus favored the mammary gland, whereas S. equorum was more common in the parlor-associated niches. Within each herd, species distribution was similar between teat apices and milking machine unit liners. In conclusion, some of the extramammary niches related to the milking process might act as infection sources for IMI-causing CNS. This study provides further evidence that the group of CNS species is comprised of environmental, opportunistic and host-adapted species which differ in ecology. Copyright © 2014 Elsevier B.V. All rights reserved.

SadA-Expressing Staphylococci in the Human Gut Show Increased Cell Adherence and Internalization

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Arif Luqman

2018-01-01

Full Text Available Summary: A subgroup of biogenic amines, the so-called trace amines (TAs, are produced by mammals and bacteria and can act as neuromodulators. In the genus Staphylococcus, certain species are capable of producing TAs through the activity of staphylococcal aromatic amino acid decarboxylase (SadA. SadA decarboxylates aromatic amino acids to produce TAs, as well as dihydroxy phenylalanine and 5-hydroxytryptophan to thus produce the neurotransmitters dopamine and serotonin. SadA-expressing staphylococci were prevalent in the gut of most probands, where they are part of the human intestinal microflora. Furthermore, sadA-expressing staphylococci showed increased adherence to HT-29 cells and 2- to 3-fold increased internalization. Internalization and adherence was also increased in a sadA mutant in the presence of tryptamine. The α2-adrenergic receptor is required for enhanced adherence and internalization. Thus, staphylococci in the gut might contribute to gut activity and intestinal colonization. : Luqman et al. examine the sadA gene and argue that it contributes to TAs. They found that neuromodulator-producing staphylococci were present in the gut of most probands. The produced neuromodulators enhanced the adherence and internalization of staphylococci to cells in culture. Keywords: adherence, aromatic amino acid decarboxylase, gut microbiota, internalization, neuromodulator, neurotransmitter, staphylococcus

Biofilm Formation by Bacteria Isolated from Intravenous Catheters

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Sina Hedayati

2015-10-01

Full Text Available Background: Reports on the association of nosocomial bacterial infections with indwelling medical devices such as intravenous catheters (IVC has increased in recent years. The potential to form biofilm on these devices seems to be the main reason for establishment of such infections. The aim of this study was to measure the potential of biofilm formation by bacterialisolates from IVCs.Methods: Seventy-one IVCs were collected from hospitalized patients in ICU, NICU, hematology and oncology wards at Taleghani Hospital from Jan 2010 to Jan 2011. The bacterial isolates were identified using the standard biochemical tests and the potential to form biofilms was determined by the microtiter plate assay method (MTP and colony morphology using Congo red agar plates (CRA.Results: Overall, 54 (71% IVCs were colonized and 76 bacteria were isolated among which, 64 (84.2% were coagulase negative staphylococci (CoNS, 3 (3.9% S. aureus, 3 (3.9% Enterococcus spp., 2 (2.6% E. coli and 4 (5.3% were miscellaneous isolates not further identified. Among the CoNS, biofilm formation was observed in 68.7% and 82.8% of bacteriausing MTP and CRA methods, respectively. S. aureus and E. coli isolates also were biofilm producers but Enterococcus and other unknown isolates were biofilm negative.Conclusions: Our results confirm that the prevalent biofilm forming bacteria on IVCs were CoNS and that was the reason for high rates of nosocomial infections.

In vitro susceptibility of gram-negative bacterial isolates to chlorhexidine gluconate.

Science.gov (United States)

Mengistu, Y; Erge, W; Bellete, B

1999-05-01

To investigate the susceptibility of clinical isolates of gram-negative bacteria to chlorhexidine gluconate. Prospective laboratory study. Tikur Anbessa Hospital, Addis Ababa, Ethiopia. Clinical specimens from 443 hospital patients. Significant number of gram negative bacteria were not inhibited by chlorhexidine gluconate (0.02-0.05%) used for antisepsis. Four hundred and forty three strains of gram-negative bacteria were isolated from Tikur Anbessa Hospital patients. Escherichia coli (31.6%) and Klebsiella pneumoniae (23%) were the most frequently isolated bacteria followed by Proteus species (13.3%), Pseudomonas species (9.2%), and Citrobacter species (6.1%). Each organism was tested to chlorhexidine gluconate (CHG), minimum inhibitory concentration (MIC) ranging from 0.0001% to 1%w/v. All Salmonella species and E. coli were inhibited by CHG, MIC or = 0.1%). Our results showed that a significant number of the gram-negative bacterial isolates were not inhibited by CHG at the concentration used for disinfection of wounds or instruments (MIC 0.02-0.05% w/v). It is therefore important to select appropriate concentration of this disinfectant and rationally use it for disinfection and hospital hygiene. Continuing follow up and surveillance is also needed to detect resistant bacteria to chlorhexidine or other disinfectants in time.

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

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

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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

Prevalence of Methicillin Resistant Staphylococcus aureus in Clinical Samples of Teerthankar Mahaveer Medical College Hospital and Research Centre (TMMCH & RC, Moradabad (UP, India

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Bina Pani Gupta

2017-06-01

Full Text Available Staphylococcus aureus is the emerging and prevalent pathogen causing serious infections in community and hospital associated diseases. S. aureus resistant to methicillin is nowadays a big and expanding problem of concern in India. Amongst the different pathogens, S. aureus is being studied for prevalence of infections and drug resistance behavior. The present study describes the dominance of Staphylococcus aureus prevalence in the clinical samples of TMU, Moradabad, India. The study showed the isolation of 450 cultures of S. aureus from different samples. Amongst which, 234 isolates of S. aureus were from pus, 164 isolates were from blood, 15 isolates were from respiratory fluid samples, 33 isolates were from urine samples and 04 isolates were from ear swabs and nasal swabs. These strains of S. aureus were screened for characteristic coagulase assay. The strains were found to be coagulase positive and coagulases negative both. It was observed that, amongst, 450 isolates of Staphylococci, 185 (41.11% strains were coagulase positive and 265 (58.88% were coagulase negative. A total of 142 (76.75% of the coagulase positive staphylococci strains shows resistance to methicillin and 202 (76.22% coagulase negative strains showed methicillin resistance. Methicillin resistance was consistent when tested with other antibiotics in coagulase positive strains but when studied about coagulase negative strains, about 12.5% strains showed sensitivity with other antibiotics although they were found resistant when checked with methicillin. It was determined that, on an average, 85 (18.88% Staphylococci strains were resistant.

Resistance to antimicrobials and biofilm formation in Staphylococcus spp. isolated from bovine mastitis in the Northeast of Brazil.

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da Costa Krewer, Carina; Santos Amanso, Evandro; Veneroni Gouveia, Gisele; de Lima Souza, Renata; da Costa, Mateus Matiuzzi; Aparecido Mota, Rinaldo

2015-03-01

Mastitis is the principal disease affecting dairy herds worldwide. The aim of the present study was to characterize phenotypic and genotypic features associated with resistance to antimicrobials in Staphylococcus spp. isolated from 2064 milk samples of 525 lactating cows in the Northeast of Brazil. Of the 218 isolates analyzed, 57.8% were characterized as Staphylococcus aureus, 28% as coagulase-positive staphylococci other than S. aureus (oCPS), and 14.2% as coagulase-negative staphylococci (CNS). The test for susceptibility to antimicrobials showed amoxicillin (32.6%) to be the less effective drug in vitro, and the multi-drug resistance (MDR) rate for beta-lactams varied from 0 to 0.75. The genotypic characterization showed that 93.1% of the samples were tested positive for the blaZ gene, while none amplified mecA. The antibiotic efflux mechanism was observed in 0.9% of isolates. The biofilm formation was found in 3.7 and 96.3% of samples, respectively, on Congo red agar and on the microplate adhesion test, while the icaD gene was present in 92.2% of Staphylococcus spp. The high frequency of blaZ gene observed in this study was associated with the resistance of most Staphylococcus spp. to one or more of the beta-lactams tested, which are routinely used in Brazilian herds for mastitis treatment. The biofilm formation was also detected in the isolates analyzed being an important characteristic for pathogenicity and antimicrobial resistance of bacteria.

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

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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.

Design and test of a novel isolator with negative resistance electromagnetic shunt damping

International Nuclear Information System (INIS)

Yan, Bo; Zhang, Xinong; Niu, Hongpan

2012-01-01

This paper proposes a negative resistance electromagnetic shunt damping vibration isolator and investigates the effectiveness of the isolator. The isolator consists of a shunt circuit and a pair of electromagnet and permanent magnets that are pasted onto a box-shaped spring. A kind of negative resistance shunt impedance is proposed to cancel the inherent resistance of the electromagnet. The electromechanical coupling coefficient and the electromagnetic damping force calculation formula are obtained by Biot–Savart’s law and Ampère’s law, respectively. A single degree of freedom system is employed to verify the performance of the proposed isolator. The governing equation is established. The performance of the proposed isolator under a half-cycle sine pulse is investigated and discussed. Experiments were carried out and the results agreed well with the numerical predictions. Both the results demonstrate that the negative resistance electromagnetic shunt damping vibration isolator could suppress vibration transmitted to the structure effectively. (paper)

[Role of proton-motive force in the conjugative DNA transport in Staphylococci].

Science.gov (United States)

Gavriliuk, V G; Vinnikov, A I

1997-01-01

Sensitivity of the conjugative process in staphylococci to the action of uncouplers of oxidative phosphorylation and inhibitors of electron transport systems have been proved, that testifies to the energy-dependent character of conjugative transport of DNA. Proceeding of the conjugation process depends upon the generation of delta microH+ on the membrane of both the donor and recipient cells. contribution of protonmotive forces to providing for the transfer of plasmids during conjugation to staphylococci has been defined.

NDM 1 Gene Carrying Gram negative Bacteria Isolated from Rats ...

African Journals Online (AJOL)

In this study, we screened 56 Gram negative bacteria comprising: 3 isolates of Enterobacter ludwigii, 30 Pseudomonas aeruginosa, 22 Proteus mirabilis, and 1 Aeromonas caviae isolated from oral cavity and rectum of rats captured from commercial poultry houses in Ibadan, Oyo State, Nigeria that were resistant to at least ...

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.

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.

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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

A daily diary study of perceived social isolation, dietary restraint, and negative affect in binge eating.

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Mason, Tyler B; Heron, Kristin E; Braitman, Abby L; Lewis, Robin J

2016-02-01

Negative affect and dietary restraint are key predictors of binge eating, yet less is known about the impact of social factors on binge eating. The study sought to replicate and extend research on the relationships between negative affect, dietary restraint, perceived social isolation and binge eating using a daily diary methodology. College women (N = 54) completed measures of dietary restraint, negative affect, perceived social isolation, and binge eating daily for 14 days. Participants completed the measures nightly each day. A series of generalized estimating equations showed that dietary restraint was associated with less binge eating while controlling for negative affect and for perceived social isolation separately. Negative affect and perceived social isolation were associated with greater binge eating while controlling for restraint in separate analyses, but only perceived social isolation was significant when modeled simultaneously. All two-way interactions between negative affect, dietary restraint, and perceived social isolation predicting binge eating were nonsignificant. This study furthers our understanding of predictors of binge eating in a nonclinical sample. Specifically, these data suggest perceived social isolation, negative affect, and dietary restraint are important variables associated with binge eating in daily life and warrant further research. Copyright © 2015 Elsevier Ltd. All rights reserved.

Plasmid-Mediated Antimicrobial Resistance in Staphylococci and Other Firmicutes.

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Schwarz, Stefan; Shen, Jianzhong; Wendlandt, Sarah; Fessler, Andrea T; Wang, Yang; Kadlec, Kristina; Wu, Cong-Ming

2014-12-01

In staphylococci and other Firmicutes, resistance to numerous classes of antimicrobial agents, which are commonly used in human and veterinary medicine, is mediated by genes that are associated with mobile genetic elements. The gene products of some of these antimicrobial resistance genes confer resistance to only specific members of a certain class of antimicrobial agents, whereas others confer resistance to the entire class or even to members of different classes of antimicrobial agents. The resistance mechanisms specified by the resistance genes fall into any of three major categories: active efflux, enzymatic inactivation, and modification/replacement/protection of the target sites of the antimicrobial agents. Among the mobile genetic elements that carry such resistance genes, plasmids play an important role as carriers of primarily plasmid-borne resistance genes, but also as vectors for nonconjugative and conjugative transposons that harbor resistance genes. Plasmids can be exchanged by horizontal gene transfer between members of the same species but also between bacteria belonging to different species and genera. Plasmids are highly flexible elements, and various mechanisms exist by which plasmids can recombine, form cointegrates, or become integrated in part or in toto into the chromosomal DNA or into other plasmids. As such, plasmids play a key role in the dissemination of antimicrobial resistance genes within the gene pool to which staphylococci and other Firmicutes have access. This chapter is intended to provide an overview of the current knowledge of plasmid-mediated antimicrobial resistance in staphylococci and other Firmicutes.

Vibration Isolation System Using Negative Stiffness(Advances in Motion and Vibration Control Technology)

OpenAIRE

水野, 毅; 高崎, 正也

2003-01-01

A new vibration isolation system using negative stiffness realized by active control technique is proposed in this paper. The serial connection of a normal spring and a suspension system with negative stiffness enables the isolation system to have low stiffness for vibration from the ground and high (theoretically infinite) stiffness against direct disturbance acting on the isolation table. A control method of realizing negative stiffness with a linear actuator is presented in an analytical f...

Identification of methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from burn patients by multiplex PCR.

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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.

Central Venous Catheter (CVC related infections: a local retrospective study

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Manuela Fresu

2008-06-01

Full Text Available Background. Central venous catheter (CVC related infection is associated with significant increases in morbidity, mortality, and health care cost.This local surveillance study was carry out to monitor the frequency of occurrence of CVC-related blood stream infections. Materials and methods. During the period January – December 2005, 226 CVC specimens were analyzed (quantitative method and microrganism identification from positive samples was performed by Vitek II. In 53 patients it was possible to compare quantitative results with those obtained from blood cultures. Results. Positive CVC samples were 125 (55% and 130 microrganisms were isolated: 109 Gram-positives (84%, 4 Gram-negatives (3%, and 17 mycetes (13%. Among pathogens collected simultaneously from CVC and blood samples, the most frequently isolated were Staphylococcus spp. (30% coagulase-negative staphylococci and 20%. S. aureus and Candida spp. (45%. In the group of patients that presented positive CVC and negative blood samples the most frequently recovered microrganisms were staphylococci. Many isolates (33% were polymicrobial. Conclusions. Catheter-related infections occurred in those patients who presented the same pathogen in both CVC and blood cultures. These infections were principally caused by staphylococci and Candida spp. On the contrary, a possible CVC contamination could be suspected when positive CVC and negative blood cultures were found.

Prevalence of mecA gene among staphylococci from clinical ...

African Journals Online (AJOL)

Background: The staphylococci have increasingly been associated with infections worldwide and anti-microbial resistance has made these versatile pathogens more recalcitrant in the hospital setting. Objectives: This study sought to investigate the occurrence and distribution of Staphylococcus species as well as determine ...

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.

Isolation of methicillin-resistant Staphylococcus pseudintermedius from breeding dogs.

Science.gov (United States)

Rota, Ada; Milani, Chiara; Drigo, Ilenia; Drigo, Michele; Corrò, Michela

2011-01-01

The overuse of antimicrobials can select resistant bacteria strains; staphylococci have the ability to become resistant to all beta-lactam antimicrobials and are a significant concern in human medicine and a growing issue for veterinary medicine. Because antimicrobials are sometimes incorrectly used in breeding kennels, the objective of the work was to assess the occurrence of methicillin-resistant coagulase-positive staphylococci in breeding dogs. The research was carried out in 13 kennels that were allotted to three categories according to the intensity of antimicrobial use. Vaginal and milk swabs were taken from 87 healthy bitches around parturition and also from multiple organs of 27 of their pups that died within the first 2 weeks. Standard bacteriological examinations were carried out and coagulase-positive staphylococci were identified. All the coagulase-positive staphylococci resulted to be Staphylococcus pseudintermedius. Susceptibility to oxacillin and the presence of the mecA gene were tested. Nine out of 89 strains (six isolated from the bitches' milk and three from dead puppies, all belonging to kennels characterized by an excessive use of antimicrobials) were multidrug-resistant, methicillin-resistant and mecA positive. Our results confirm that excessive use of antimicrobials entails the risk of selecting resistant staphylococci strains. Our data also indicate that the bacterial flora of healthy dogs belonging to specific populations may act as a reservoir of resistance genes. Copyright © 2011 Elsevier Inc. All rights reserved.

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

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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

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

Heifers infected with coagulase-negative staphylococci in early lactation have fewer cases of clinical mastitis and higher milk production in their first lactation than noninfected heifers.

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Piepers, S; Opsomer, G; Barkema, H W; de Kruif, A; De Vliegher, S

2010-05-01

Intramammary infections (IMI) in recently calved dairy heifers are more common than was formerly believed but their relevance for future performance has been studied only rarely. In the present study, the association between the IMI status of fresh heifers and their subsequent udder health, milk production, and culling in first lactation was explored. Quarter milk samples were collected between 1 and 4 d in milk (DIM) and between 5 and 8 DIM from 191 dairy heifers in 20 dairy herds for bacteriological culturing and somatic cell count (SCC) analysis. Monthly milk recording data including composite milk SCC and test-day milk yield (MY) were obtained for the first 285 DIM or until culling. Farmer-recorded clinical mastitis cases were available. Data were analyzed using mixed models and survival analysis. Approximately 80% of the fresh heifers (79.8%) had at least one culture-positive quarter. Coagulase-negative staphylococci (CNS) were the most frequently isolated pathogens (72%), followed by esculin-positive streptococci (4.6%) and Staphylococcus aureus (3.5%). Overall geometric mean SCC at quarter level decreased between the first and second samplings from 348,000 to 116,000 cells/mL. Heifers infected with CNS had an intermediate average test-day SCC (84,000 cells/mL) during the first 285 DIM compared with noninfected heifers (53,000 cells/mL) and heifers infected with major pathogens (195,000 cells/mL). Heifers infected with major pathogens had a much lower average daily MY (18.3kg) during first lactation compared with noninfected animals (21.3kg). That CNS-infected heifers out-produced their noninfected counterparts could be at least partially explained by their significantly lower incidence of clinical mastitis (incidence risk 3.6 vs. 21.0%) during first lactation compared with noninfected heifers. We conclude that although CNS cause the majority of IMI in heifers around calving, they should not be a reason for serious concern. Copyright 2010 American Dairy

Persistence of mixed staphylococci assemblages following disinfection of hospital room surfaces.

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Sigler, V; Hensley, S

2013-03-01

The distribution of staphylococcal assemblages on surfaces in hospital rooms was assessed before and after daily disinfection with quaternary ammonia products. DNA was extracted from enrichment cultures of bacteria, which were swabbed from each of nine surface types, and subjected to analysis by staphylococci-specific, denaturing gradient gel electrophoresis. A genetic marker for Staphylococcus epidermidis/kloosii was detected on all surface types before and after cleaning, whereas markers for Staphylococcus aureus and Staphylococcus lugdunensis were detected on five surface types. Overall, genetic makers for several staphylococci known to colonize and infect humans remained ubiquitous in each room following daily disinfection practices. Copyright © 2013 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

Presence of superantigen genes and antimicrobial resistance in Staphylococcus isolates obtained from the uteri of dairy cows with clinical endometritis.

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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. British Veterinary Association.

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

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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.

HEALTH AND HYGIENIC CONDITIONS OF EWE'S MILK PROCESSING FROM THE ASPECT OF FOOD SAFETY

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Jana Pukáčová

2010-07-01

Full Text Available Totally, 47 strains of S. aureus and 578 coagulase negative staphylococci were detected in samples from raw ewe milk. The 35 out 47 isolates of S. aureus from ewe milk were positive for the presence of staphylococcal enterotoxin genes: sea(4 %, sec (48 % a sed (48 %. Staphylococcus epidermis (33.04%, Staphylococcus caprae (21.28% were more prevalent. Staphylococcus chromogenes (7.44 %, Staphylococcus hominis (7.09%, Staphylococcus xylosus (6,92 %, a Staphylococcus warneri (6.40 % were isolated also in ewes milk. Staphylococcus haemolyticus (3.11 %, Staphylococcus capitis (2.94 %, Staphylococcus simulans (2.08 % and Staphylococcus saprophyticus (1.73 % were isolated very rarely from the taken individual milk ewe samples. Sporadically, only in few cases, the others coagulase negative staphylococci were isolated (Staphylococcus cohnii cohnii, Staphylococcus sciuri, Staphylococcus closii, Staphylococcus lugdunensis, Staphylococcus auricularis and Staphylococcus equorum.   doi:10.5219/24

Utilization of Cypermethrin by bacteria isolated from irrigated soils ...

African Journals Online (AJOL)

Soil bacteria capable of utilizing Cypermethrin as a source of carbon were isolated using enrichment technique. The bacteria were Psuedomonas aeruginosa, Serratia spp Micrococcus sp, Staphylococci and Streptococcus sp. Growth of P. aeruginosa was determined in the presence of 1:106 and 1:105 Cypermethrin in ...

A novel magnetorheological elastomer isolator with negative changing stiffness for vibration reduction

International Nuclear Information System (INIS)

Yang, J; Sun, S S; Li, W H; Alici, G; Du, H; Deng, H X

2014-01-01

Magneto-rheological elastomers (MREs) have attracted notable credits in the development of smart isolators and absorbers due to their controllable stiffness and damping properties. For the purpose of mitigating unwanted structural and/or machinery vibrations, the traditional MRE-based isolators have been generally proven effective because the MR effect can increase the stiffness when the magnetic field is strengthened. This study presents a novel MRE isolator that experienced reduced stiffness when the applied current was increased. This innovative work was accomplished by applying a hybrid magnet (electromagnet and permanent magnets) onto a multilayered MRE structure. To characterise this negative changing stiffness concept, a multilayered MRE isolator with a hybrid magnet was first designed, fabricated and then tested to measure its properties. An obvious reduction of the effective stiffness and natural frequency of the proposed MRE isolator occurred when the current was continuously adjusted. This device could also work as a conventional MRE isolator as its effective stiffness and natural frequency also increased when a negative current was applied. Further testing was carried out on a one-degree-of-freedom system to assess how effectively this device could isolate vibration. In this experiment, two cases were considered; in each case, the vibration of the primary system was obviously attenuated under ON-OFF control logic, thus demonstrating the feasibility of this novel design as an alternative adaptive vibration isolator. (paper)

SadA-Expressing Staphylococci in the Human Gut Show Increased Cell Adherence and Internalization.

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Luqman, Arif; Nega, Mulugeta; Nguyen, Minh-Thu; Ebner, Patrick; Götz, Friedrich

2018-01-09

A subgroup of biogenic amines, the so-called trace amines (TAs), are produced by mammals and bacteria and can act as neuromodulators. In the genus Staphylococcus, certain species are capable of producing TAs through the activity of staphylococcal aromatic amino acid decarboxylase (SadA). SadA decarboxylates aromatic amino acids to produce TAs, as well as dihydroxy phenylalanine and 5-hydroxytryptophan to thus produce the neurotransmitters dopamine and serotonin. SadA-expressing staphylococci were prevalent in the gut of most probands, where they are part of the human intestinal microflora. Furthermore, sadA-expressing staphylococci showed increased adherence to HT-29 cells and 2- to 3-fold increased internalization. Internalization and adherence was also increased in a sadA mutant in the presence of tryptamine. The α2-adrenergic receptor is required for enhanced adherence and internalization. Thus, staphylococci in the gut might contribute to gut activity and intestinal colonization. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

A novel MSCRAMM sub-family in Coagulase negative staphylococcal species

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Srishtee eArora

2016-04-01

Full Text Available Coagulase negative staphylococci are important opportunistic pathogens. Staphylococcus epidermidis, a coagulase negative staphylococcus, is the third leading cause of nosocomial infections in the US. Surface proteins like Microbial Surface Components Recognizing Adhesive Matrix Molecules (MSCRAMMs are major virulence factors of pathogenic gram positive bacteria. Here, we identified a new chimeric protein; SesJ in S. epidermidis, which represents a prototype of a new subfamily of MSCRAMMs. Structural predictions show that SesJ has structural features characteristic of a MSCRAMM along with a N-Terminal repeat region and an Aspartic acid containing repeat region, features that have not been previously observed in staphylococcal MSCRAMMs but have been found in other surface proteins from gram positive bacteria. We identified and analyzed structural homologs of SesJ in three other coagulase negative staphylococci. These homologs of SesJ have an identical structural organization but varying sequence identities within the domains. Using flow cytometry, we also show that SesJ is expressed constitutively on the surface of a representative S. epidermidis strain, from early exponential to stationary growth phase. Thus SesJ is positioned to interact with protein targets in the environment and play a role in S. epidermidis virulence.

Use of a primary isolation medium for recovery of methicillin-resistant Staphylococcus aureus.

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Van Enk, R A; Thompson, K D

1992-01-01

Clinical specimens frequently contain methicillin-resistant Staphylococcus aureus (MRSA) isolates in low numbers or mixed with methicillin-susceptible staphylococci, which can obscure MRSA on nonselective media. By using an oxacillin-containing mannitol-salt-based selective and differential medium on 936 respiratory specimens, we recovered 45% more MRSA isolates (29 versus 20) than on nonselective media alone.

The effect of clary sage oil on staphylococci responsible for wound infections.

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Sienkiewicz, Monika; Głowacka, Anna; Poznańska-Kurowska, Katarzyna; Kaszuba, Andrzej; Urbaniak, Anna; Kowalczyk, Edward

2015-02-01

The spreading of bacterial antibiotic resistance among clinical strains of pathogenic bacteria has made investigators to search for other active antibacterial agents which could provide a valuable complement to the existing therapies. To determine the antibacterial activity of clary sage oil (Salvia sclarea L.) against Staphylococcus clinical strains which were isolated from patients with wound infections. A comprehensive evaluation of Staphylococcus clinical strain resistance to antibiotics was performed. The constituents of clary sage oil were assayed by GC-FID-MS analysis. The minimal inhibitory concentration (MIC) of the tested essential oil against staphylococci by the micro-dilution broth method was determined. The clary sage oil was active against Staphylococcus aureus, S. epidermidis and S. xylosus with MIC values ranging from 3.75 to 7.00 µl/ml. The results of the in vitro tests encourage to use formulations containing sage oil as the active natural antimicrobial agent. Because of its antimicrobial properties clary sage oil may be applied to treat wounds and skin infections.

Significance of coagulase negative Staphylococcus from blood cultures: persisting problems and partial progress in resource constrained settings.

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Sidhu, Shailpreet K; Malhotra, Sita; Devi, Pushpa; Tuli, Arpandeep K

2016-12-01

Coagulase negative Staphylococcus (CoNS) is frequently isolated from blood cultures but their significance is difficult to interpret. CoNS bacteria which are often previously dismissed as culture contaminants are attracting greater importance as true pathogens in the past decades. Clinical evaluation of these isolates suggests that although there is a relative increase of CoNS associated bloodstream infections in recent years, the microorganisms still remain the most common contaminants in blood cultures. The objective of this study was to determine the significance of CoNS isolated from blood cultures. A retrospective study was conducted to evaluate the rate of contamination in blood cultures in a tertiary care hospital. The paired specimens of blood were cultured using conventional culture methods and the isolates of coagulase negative staphylococci were identified by standard methodology. Clinical data, laboratory indices, microbiological parameters and patient characteristics were analyzed. Of 3503 blood samples, CoNS were isolated from blood culture of 307 patients (8.76%). The isolates were reported as true pathogens of bloodstream infections in only 74 out of 307 cases (24.1%). In the vast majority, 212 of 307 (69.0%), they were mere blood culture contaminants and reported as insignificant/contaminant. Determining whether a growth in the blood culture is a pathogen or a contaminant is a critical issue and multiple parameters have to be considered before arriving at a conclusion. Ideally, the molecular approach is for the most part a consistent method in determining the significant isolates of CoNS. However, in countries with inadequate resources, species identification and antibiogram tests are recommended when determining significance of these isolates.

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

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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

Prevalence and antimicrobial resistance pattern of coagulase ...

African Journals Online (AJOL)

Prevalence and antimicrobial resistance pattern of coagulase negative Staphylococci isolated from pigs and in-contact humans in Jos Metropolis, Nigeria. ... (53/401) of the isolates were CoNS species based on confirmatory test with Microgen biochemical kit and were further subjected to antibiotic susceptibility testing.

Comparison Predominant Oral micro-flora in Subjects with and without Complete Denture Referred to Yazd Dentistry Department

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Abbas Ali Jafari

2014-11-01

Results: The non-aureus staphylococcus and alpha-hemolytic streptococci showed the highest positive culture among the isolated microorganisms in both groups, whereas beta hemolytic streptococci showed the least percent of isolated microorganism in both groups. The higher density of non-aureus Staphylococci, α-hemolitic Streptococci, Gram negative cocobasillus, non-pathogenic Neisseria, Candida and Corynebacterium were recovered from oral samples of denture users in compare with dentate subjects (P= 0.0001. There was also seen a statistical significant correlation between the number of isolated microorganisms and the duration of denture utilization in denture users (P=0.013. Conclusion: Results of the present study showed that complete denture can be act as a predisposal factor in overgrowing of several oral micro-flora particularly Candida, non-aureus Staphylococci, α-hemolytic streptococci, gram negative cocobacillus, non-pathogenic Neisseria, and Corynebacterium, which emphasized the users denture hygine.

The analysis of milk components and pathogenic bacteria isolated from bovine raw milk in Korea.

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Park, Y K; Koo, H C; Kim, S H; Hwang, S Y; Jung, W K; Kim, J M; Shin, S; Kim, R T; Park, Y H

2007-12-01

Bovine mastitis can be diagnosed by abnormalities in milk components and somatic cell count (SCC), as well as by clinical signs. We examined raw milk in Korea by analyzing SCC, milk urea nitrogen (MUN), and the percentages of milk components (milk fat, protein, and lactose). The associations between SCC or MUN and other milk components were investigated, as well as the relationships between the bacterial species isolated from milk. Somatic cell counts, MUN, and the percentages of milk fat, protein, and lactose were analyzed in 30,019 raw milk samples collected from 2003 to 2006. The regression coefficients of natural logarithmic-transformed SCC (SCCt) on milk fat (-0.0149), lactose (-0.8910), and MUN (-0.0096), and those of MUN on milk fat (-0.3125), protein (-0.8012), and SCCt (-0.0671) were negative, whereas the regression coefficient of SCCt on protein was positive (0.3023). When the data were categorized by the presence or absence of bacterial infection in raw milk, SCCt was negatively associated with milk fat (-0.0172), protein (-0.2693), and lactose (-0.4108). The SCCt values were significantly affected by bacterial species. In particular, 104 milk samples infected with Staphylococcus aureus had the highest SCCt (1.67) compared with milk containing other mastitis-causing bacteria: coagulase-negative staphylococci (n = 755, 1.50), coagulase-positive staphylococci (except Staphylococcus aureus; n = 77, 1.59), Streptococcus spp. (Streptococcus dysgalactiae, n = 37; Streptococcus uberis, n = 12, 0.83), Enterococcus spp. (n = 46, 1.04), Escherichia coli (n = 705, 1.56), Pseudomonas spp. (n = 456, 1.59), and yeast (n = 189, 1.52). These results show that high SCC and MUN negatively affect milk components and that a statistical approach associating SCC, MUN, and milk components by bacterial infection can explain the patterns among them. Bacterial species present in raw milk are an important influence on SCC in Korea.

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

International Nuclear Information System (INIS)

Le, Thanh Danh; Ahn, Kyoung Kwan

2012-01-01

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

Survey of Staphylococcus isolates among hospital personnel, environment and their antibiogram with special emphasis on methicillin resistance

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Shobha K

2005-01-01

Full Text Available The objective of this study was to find the prevalence of Staphylococcus spp. carriage among hospital personnel and hospital environment and their antibiogram with special emphasis on methicillin resistance. A total of 205 samples from hospital personnel and environment were collected from casualty, oncology and multidisciplinary cardiac unit ward of Kasturba Medical College Hospital, Manipal. Samples were collected using sterile cotton wool swabs and inoculated into brain heart infusion broth. Subcultures were done onto blood agar and MacConkey′s agar. Isolates were identified by standard methods up to species level. Antimicrobial susceptibility test was performed according to standardized disc diffusion Kirby-Bauer method. Each of the isolates was screened for methicillin resistance using oxacillin disc on Mueller Hinton agar plate followed by MIC for methicillin and cefoxitin susceptibility test by disc diffusion method. Sixty five out of 205 strains (31.7% were Staphylococcus spp. and all of them were coagulase negative. Most of the strains belonged to S.epidermidis 49.23%(32/65 followed by S. saprophyticus 26.15%(17/65. Maximum isolates of S.epidermidis were from anterior nares 28.12%(9/32 strains of S.epidermidis . Highest number of methicillin resistant coagulase negative strains (3/9, 33.33% were isolated from stethoscope of multidisciplinary cardiac unit ward followed by carriers in the anterior nares (2/9, 22.22%. Methicillin resistant coagulase negative staphylococci are prevalent in anterior nares of hospital personnel and in the hospital environment thereby providing a definite source for hospital acquired infection. All isolates were sensitive to vancomycin, ciprofloxacin and amikacin.

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

Science.gov (United States)

Souza Antunes, Ana Lúcia; Secchi, Carina; Reiter, Keli Cristine; Rodrigues Perez, Leandro Reus; Peixoto de Freitas, Ana Lúcia; Alves d'Azevedo, Pedro

2007-09-01

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 microg) and cefoxitin (30 microg) 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.

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.

Carriage of Staphylococcus species in the veterinary visiting dog population in mainland UK: molecular characterisation of resistance and virulence.

Science.gov (United States)

Wedley, Amy L; Dawson, Susan; Maddox, Thomas W; Coyne, Karen P; Pinchbeck, Gina L; Clegg, Peter; Jamrozy, Dorota; Fielder, Mark D; Donovan, David; Nuttall, Tim; Williams, Nicola J

2014-05-14

This study investigated the prevalence of nasal carriage of staphylococci in dogs and determined the characteristics of the isolates. A total of 724 dogs from 87 veterinary practices across the mainland UK were screened for carriage of Staphylococcus spp. All isolates were examined for meticillin resistance (MR) and the presence of the mecA gene investigated in those isolates showing resistance. All coagulase-positive staphylococci and MR coagulase-negative staphylococci (MRCoNS) were subjected to antimicrobial susceptibility testing. Spa typing and DNA microarray analysis of resistance and virulence genes was carried out on all MR S. aureus (MRSA) and a subset of meticillin susceptible S. aureus (MSSA). Staphylococci were isolated from 399 (55.1%) of the dogs; only seven (1%) carried MRSA, all of which were identified as the dominant UK healthcare-associated strain (EMRSA-15, ST22). MSSA was identified in 47 (6.5%) dogs, the sequence types of which have been suggested as precursors to successful MRSA clones. Forty (5.5%) dogs carried MRCoNS, while no dogs carried MR S. pseudintermedius, although this is increasingly reported in mainland Europe. Resistance to antimicrobials among the isolates varied between species, with multidrug resistance (MDR) in 87.5% of MRCoNS and 21.8% of coagulase positive staphylococci. Microarray analysis of MRSA and a subset of MSSA isolates identified numerous virulence genes associated with pathogenesis, which are commonly identified in isolates of human origin. However, no isolates carried Panton-Valentine leukocidin (PVL) genes. This study suggests that MRSA carriage is low in the vet visiting dog population, but there is a diverse range of virulence and resistance determinants in canine S. aureus and MRCoNS isolates. Copyright © 2014 Elsevier B.V. All rights reserved.

Species Distribution and Antibiotic Resistance in Coagulase ...

African Journals Online (AJOL)

Purpose: The antimicrobial susceptibility of 149 coagulase-negative staphylococci (CoNS) isolates from faecal samples of children in Ile-Ife, Nigeria, was evaluated in order to determine their contribution to antimicrobial resistance in the community. Methods: The isolates were identified to the species level by conventional ...

Distribution of non-aureus staphylococci species in udder quarters with low and high somatic cell count, and clinical mastitis.

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Condas, Larissa A Z; De Buck, Jeroen; Nobrega, Diego B; Carson, Domonique A; Roy, Jean-Philippe; Keefe, Greg P; DeVries, Trevor J; Middleton, John R; Dufour, Simon; Barkema, Herman W

2017-07-01

The effect of non-aureus staphylococci (NAS) in bovine mammary health is controversial. Overall, NAS intramammary infections (IMI) increase somatic cell count (SCC), with an effect categorized as mild, mostly causing subclinical or mild to moderate clinical mastitis. However, based on recent studies, specific NAS may affect the udder more severely. Some of these apparent discrepancies could be attributed to the large number of species that compose the NAS group. The objectives of this study were to determine (1) the SCC of quarters infected by individual NAS species compared with NAS as a group, culture-negative, and major pathogen-infected quarters; (2) the distribution of NAS species isolated from quarters with low SCC (mastitis; and (3) the prevalence of NAS species across quarters with low and high SCC. A total of 5,507 NAS isolates, 3,561 from low SCC quarters, 1,873 from high SCC quarters, and 73 from clinical mastitis cases, were obtained from the National Cohort of Dairy Farms of the Canadian Bovine Mastitis Research Network. Of quarters with low SCC, high SCC, or clinical mastitis, 7.6, 18.5, and 4.3% were NAS positive, respectively. The effect of NAS IMI on SCC was estimated using mixed-effect linear regression; prevalence of NAS IMI was estimated using Bayesian analyses. Mean SCC of NAS-positive quarters was 70,000 cells/mL, which was higher than culture-negative quarters (32,000 cells/mL) and lower than major pathogen-positive quarters (129,000 to 183,000 cells/mL). Compared with other NAS species, SCC was highest in quarters positive for Staphylococcus capitis, Staphylococcus gallinarum, Staphylococcus hyicus, Staphylococcus agnetis, or Staphylococcus simulans. In NAS-positive quarters, Staphylococcus xylosus (12.6%), Staphylococcus cohnii (3.1%), and Staphylococcus equorum (0.6%) were more frequently isolated from quarters with low SCC than other NAS species, whereas Staphylococcus sciuri (14%) was most frequently isolated from clinical mastitis cases

Retrospective Analysis of Blood Stream Infections and Antibiotic Susceptibility Pattern of Gram Negative Bacteria in a Tertiary Care Cancer Hospital

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Radha Rani D

2017-12-01

Full Text Available Background: Bacterial bloodstream infections are important causes of morbidity and mortality globally. The aim of the present study was to determine the bacterial profile of bloodstream infections and their antibiotic susceptibility pattern among the clinically diagnosed cases of sepsis in cancer patients. Methods: In the present study, etiological and antimicrobial susceptibility profile of blood cultures over a period of 1 year at a tertiary cancer care hospital was done. Blood culture positive isolates were identified using standard microbiological methods and by Fully automated BD Phoenix 100. The antibiotic susceptibility pattern of the organisms was performed by Kirby-Bauer disc diffusion method and MIC (Minimum inhibitory concentration was done by Fully automated BD Phoenix 100. Results: There were 1178 blood culture samples, of which 327 (27.7% were identified to be culture positive. Out of 327 positive cultures, 299 (91.4% showed bacterial growth, Gram negative were 161 (53.8% and Gram positive were 138 (46.1%. Candida species were isolated from 13 (3.97% of positive samples and 15 samples showed contamination. The most common Gram-negative isolate was. Escherichia coli (37.80% and Gram-positive isolate was coagulasenegative staphylococci (52.80%. Escherichia coli showed highest sensitivity to amikacin (83.60% and sensitivity to piperacillin+ tazobactum and cefaperazone+sulbactam was 54.09% and 52.45% respectively. High degree of resistance was found to cephalosporins and levofloxacin. Conclusion: The results indicate high level of antimicrobial resistance among Gram negative bacilli in septicemic patients. The results warrant continuous monitoring of antimicrobial pattern so as to build geographical epidemiological data.

Antimicrobial susceptibility patterns and CTX-M β-lactamase producing clinical isolates from burn patients in Islamabad, Pakistan

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Mubbashir Hussain

2017-08-01

Full Text Available Objective: To evaluate the prevalence of extended spectrum beta-lactamases (ESBL in clinical isolates from burn patients using phenotypic and genotypic analyses. Methods: During 2015–2016, a total of 126 samples were collected at a tertiary care hospital, Islamabad. Antibiotic sensitivity and ESBL prevalence were evaluated according to the Clinical Laboratory and Standards Institute, and molecular analysis of the CTX-M type ESBL gene was performed in 225 bacterial isolates from these samples. Results: The most prevalent bacterial species were Escherichia coli (28.4%, Pseudomonas aeruginosa (22.2%, Staphylococcus aureus (19.6%, Klebsiella pneumoniae (16.4%, and coagulase-negative staphylococci (13.3%. Of the 225 bacterial isolates, 89 (39.5% were found to be ESBL producers. The isolates were highly susceptible to meropenem (88% and imipenem (84%, followed by the aminoglycoside amikacin (81%. Molecular epidemiology of the ESBL isolates indicated 19% prevalence of CTX-M. Resistance to antibiotics was exhibited by 28% isolates. Conclusions: In the present study, bacteria such as P. aeruginosa, K. pneumoniae, S. aureus, and E. coli isolated from burn patients exhibited resistance to one or more antibiotics and produced large amounts of ESBL. Further studies are needed to investigate the virulence and epidemiology of CTX-M type ESBL in clinical isolates from burn patients.

Subclinical mastitis in sheep: Causes and their sensitivity to antibiotics

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Vakanjac Slobodanka

2010-01-01

Full Text Available The California mastitis test (CMT was used to examine the milk of 6609 sheep, actually, from 13218 mammary complexes. A total of 986 milk samples were found to contain an increased number of somatic cells, and causes of mastitis were isolated from 910 (92.3% of these samples. The most frequently isolated microorganisms were: E. coli isolated from 189 (20.76% samples, Micrococcus sp from 186 (20.43% samples, Bacillus sp. from 177 (19.45%, Staphylococcus aureus from 172 (18.9% samples, and Proteus sp. from 121 (13.29% samples of sheep milk. Coagulase-negative staphylococci (CNS were isolated in a much smaller number, from 25 (2.74% samples, Streptococcus sp. was isolated from 19 (2.08% samples, Pseudomonas sp. from 14 (1.53% samples, haemolytic E. coli was isolated from only 6 (0.65% samples, and Streptococcus agalactiae from only one sample. Among the 25 isolates of coagulase-negative staphylococci, 16 (64% isolates were identified as Staphylococcus chromogenes, and 9 (36% isolates as Staphylococcus simulans. The sensitivity of the isolated causes of mastitis to antibiotics was examined using the disc diffusion method. .

[The HVR genotypes and their relationship with the resistance of methicillin-resistant staphylococci].

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Liao, F; Fan, X; Lü, X; Feng, P

2001-06-01

To investigate the HVR-PCR genotype of methicillin-resistant Staphylococci in local hospitals and compare it with the antibiograms, with aview to selecting effective antibacterial agents, moreover, to discuss preliminarily its role in molecular epidemiology. The minimal inhibitory concentrations(MICs) of 86 MRSA, 10 MRSE(Mc'S. epidemidis), 5 MSSE(Mc'S. epidemidis), 8 MRSH(Mc'S. haemolyticus) and 5 MSSH(Mc'S. haemolyticus) clinical isolates collected from 4 local hospitals were tested by serial two-fold agar dilution method; their DNA were extracted by moved basic lytic method, whose polymerase chain reaction(PCR) products amplified, based on the size of mec-associated hypervariable region(HVR) were analyzed by PAG vertical and agarose gel electrophoresis. MRSA, MRSE and MRSH were grouped into 4, 3 and 2 HVR genotypes respectively according to the size of the PCR products. The PCR products amplified from 9 of 10 MRSE isolates were the same as the products amplified from MRSA isolates. MRSA strains in this study were mainly HVR genotypes A and D, which accounted for 52.32% and 39.53%; Genotypes B and C were the most multi-drug resistant, but genotype D was multi-sensitive. The I genotype of MRSE was multi-drug resistant, but its genotype III was multi-drug sensitive. The genotype a of MRSH was more resistant than genotype b. These results suggest that HVR-PCR genotype method is an easy and fast method for epidemiological investigation of nosocomial infections caused by MRSA, and it is helpful for clinical selection of antibacterial agents. This method can compare the mec determinants of MRSA and Mc'CNSt isolates and hence to search for the origin of the mec determinant.

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

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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.

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

Central venous catheter-related bloodstream infections in cancer patients

International Nuclear Information System (INIS)

Butt, T.; Afzal, R.K.; Ahmad, R.N.; Hussain, I.; Anwar, M.

2004-01-01

Objective: To determine the frequency of central venous catheter-related bloodstream infections (CR-BSI) in cancer patients and the antimicrobial susceptibility pattern of the isolates. Subjects and Methods: Cancer patients requiring short or long-term central venous catheterization at the time of admission or thereafter were included. Catheter tips on removal were cultured quantitatively; specimens of blood and pus were cultured qualitatively. Isolates were identified and antimicrobial susceptibility testing was performed by standard techniques. Results: Eighty-nine patients were included in the study. The frequency of CR-BSI was 17%. Out of the 19 organisms isolated, 10 (53%) were Gram-positive cocci, 8 (42%) were Gram-negative rods and 1 (5%) was a fungus. Coagulase negative staphylococci (27%) were the predominant pathogens. Among the staphylococci, 46% of the isolates were methicillin-resistant. All Gram-positive isolates were susceptive to glycopeptides. Gram-negative rods were resistant to most of the commonly used antimicrobial groups. Conclusion: Central venous catheter is an important source of bloodstream infections in cancer patients. Most of the infections are caused by Gram-positive cocci. Rigorous infection control measures and continuous surveillance is required to curb the frequency of these infections. (author)

Increases of Antibiotic Resistance in Excessive Use of Antibiotics in Smallholder Dairy Farms in Northern Thailand

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W. Suriyasathaporn

2012-09-01

Full Text Available Antibiotic resistance patterns of bacterial isolates from both quarter teat-tip swabs and their quarter milk samples were evaluated in smallholder dairy farms in northern Thailand with excessive use of antibiotics (HIGH compared with normal use (NORM. Results from teat-tip swab samples showed that the percentage of Bacillus spp. resistance to overall antibiotics was significantly lower in the NORM group than that of the HIGH group, whereas, the resistance percentage of coagulase-negative staphylococci in the NORM group was higher than that of the HIGH one. The overall mastitis-causing bacteria isolated from milk samples were environmental streptococci (13.8%, coagulase-negative staphylococci (9.9%, Staphylococcus aureus (5.4%, and Corynebacterium bovis (4.5%. Both staphylococci and streptococci had significantly higher percentages of resistance to cloxacillin and oxacillin in the HIGH group when compared to the NORM one. An occurrence of vancomycin-resistant bacteria was also observed in the HIGH group. In conclusion, the smallholder dairy farms with excessive use of antibiotics had a higher probability of antibiotic-resistant pattern than the farms with normal use.

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.

Increasing trend of community-acquired methicillin-resistant: Staphylococcal carriers: An alarming bell for urgent measures

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Poongodi Lakshmi Santhana Kumarasamy

2015-01-01

Full Text Available Background: An increase in the incidence of infections caused by community-associated-methicillin resistant Staphylococcus aureus (MRSA has been reported. Hence, the knowledge of resistance pattern of these isolates is a precondition for alleviating emerging antibiotic resistance and devising better treatment strategies Aim: To find out the prevalence of community-acquired methicillin-resistant staphylococcal strains from nasal carriers. Materials and Methods: A total of 352 nasal swabs collected during routine health checkup were analyzed. Results: Of the 58 (16% staphylococci isolated, 32 (55% were S. aureus and 26 (45% were coagulase-negative staphylococci (CoNS. Methicillin resistance was observed in 7 (22% of staphylococci aureus and 11 (42% of CoNS. "D test" was positive in 1 (14% MRSA, 2 (8% methicillin-susceptible S. aureus and 2 (8% methicillin resistant-CoNS. Conclusion: Effective implementation of the antibiotic policy along with measures like hand wash, isolation of patients will reduce the incidence of resistance.

Evaluation of pet contact as a risk factor for carriage of multidrug-resistant staphylococci in nursing home residents.

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Gandolfi-Decristophoris, Paola; De Benedetti, Anna; Petignat, Christiane; Attinger, Monica; Guillaume, Jan; Fiebig, Lena; Hattendorf, Jan; Cernela, Nicole; Regula, Gertraud; Petrini, Orlando; Zinsstag, Jakob; Schelling, Esther

2012-03-01

Pets, often used as companionship and for psychological support in the therapy of nursing home residents, have been implicated as reservoirs for antibiotic-resistant bacteria. We investigated the importance of pets as reservoirs of multidrug-resistant (MDR) staphylococci in nursing homes. We assessed the carriage of MDR staphylococci in pets and in 2 groups of residents, those living in nursing homes with pets and those living without pet contacts. We collected demographic, health status, and human-pet contact data by means of questionnaires. We assessed potential bacteria transmission pathways by investigating physical resident-to-pet contact. The observed prevalence of MDR staphylococci carriage was 84/229 (37%) in residents living with pets and 99/216 (46%) in those not living with pets (adjusted odds ratio [aOR], 0.6; 95% confidence interval [CI], 0.4-0.9). Active pet contact was associated with lower carriage of MDR staphylococci (aOR, 0.5; 95% CI, 0.4-0.8). Antibiotic treatment during the previous 3 months was associated with significantly increased risk for MDR carriage in residents (aOR, 3.1; 95% CI, 1.8-5.7). We found no evidence that the previously reported benefits of pet contact are compromised by the increased risk of carriage of MDR staphylococci in residents associated with interaction with these animals in nursing homes. Thus, contact with pets, always under good hygiene standards, should be encouraged in these settings. Copyright © 2012 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.

Comparative assessment of antibiotic susceptibility of coagulase-negative staphylococci in biofilm versus planktonic culture as assessed by bacterial enumeration or rapid XTT colorimetry.

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Cerca, Nuno; Martins, Silvia; Cerca, Filipe; Jefferson, Kimberly K; Pier, Gerald B; Oliveira, Rosário; Azeredo, Joana

2005-08-01

To quantitatively compare the antibiotic susceptibility of biofilms formed by the coagulase-negative staphylococci (CoNS) Staphylococcus epidermidis and Staphylococcus haemolyticus with the susceptibility of planktonic cultures. Several CoNS strains were grown planktonically or as biofilms to determine the effect of the mode of growth on the level of susceptibility to antibiotics with different mechanisms of action. The utility of a new, rapid colorimetric method that is based on the reduction of a tetrazolium salt (XTT) to measure cell viability was tested by comparison with standard bacterial enumeration techniques. A 6 h kinetic study was performed using dicloxacillin, cefazolin, vancomycin, tetracycline and rifampicin at the peak serum concentration of each antibiotic. In planktonic cells, inhibitors of cell wall synthesis were highly effective over a 3 h period. Biofilms were much less susceptible than planktonic cultures to all antibiotics tested, particularly inhibitors of cell wall synthesis. The susceptibility to inhibitors of protein and RNA synthesis was affected by the biofilm phenotype to a lesser degree. Standard bacterial enumeration techniques and the XTT method produced equivalent results both in biofilms and planktonic assays. This study provides a more accurate comparison between the antibiotic susceptibilities of planktonic versus biofilm populations, because the cell densities in the two populations were similar and because we measured the concentration required to inhibit bacterial metabolism rather than to eradicate the entire bacterial population. While the biofilm phenotype is highly resistant to antibiotics that target cell wall synthesis, it is fairly susceptible to antibiotics that target RNA and protein synthesis.

Molecular Typing and Antimicrobial Susceptibility of Methicillin-Resistant Staphylococcus aureus Isolated from Bovine Milk in Tanzania.

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Mohammed, Jibril; Ziwa, Michael Henry; Hounmanou, Yaovi Mahuton Gildas; Kisanga, Adela; Tuntufye, Huruma Nelwike

2018-01-01

Methicillin-resistant Staphylococcus aureus (MRSA) in raw milk can be transmitted from animals to humans, and in Tanzania raw milk is sold in local markets and consumed as purchased. This study was performed to determine the molecular characteristics and antimicrobial susceptibility pattern of MRSA strains isolated from raw bovine milk sold at local markets in Tanzania. A total of 117 raw milk samples were cultured on Baird-Parker medium to isolate S. aureus and PCR was used for amplification of gltB gene for S. aureus identification and the presence of mecA gene for methicillin-resistant strains. Coagulase-negative (CN) S. aureus were reconfirmed using tube coagulase, DNase, and API Staph tests. MRSA isolates were spa typed whereas antimicrobial susceptibility testing was performed by the disc diffusion method. Forty-six coagulase positives (CP) and two CN S. aureus were identified. Most strains were resistant to penicillin (72%), and 3 isolates: 2 CN S. aureus and 1 coagulase-negative Staphylococci (CNS), were phenotypically resistant to vancomycin, oxacillin, and cefoxitin and were confirmed to carry mecA. Resistance to clindamycin, trimethoprim-sulfamethoxazole, and tetracycline was 23.9%, 30.4%, and 41.3%, respectively. Twelve isolates exhibited multidrug resistance; however, only one mecA positive strain among the three was typeable and belonged to spa type t2603. This study reports for the first time the presence of CN variant of MRSA, which was assigned the spa type t2603, and the presence of multidrug resistant S. aureus isolates from bovine milk in Morogoro, Tanzania.

Bacterial Isolates and Their Antimicrobial Susceptibility Patterns of Wound Infections among Inpatients and Outpatients Attending the University of Gondar Referral Hospital, Northwest Ethiopia

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Aynalem Mohammed

2017-01-01

Full Text Available Background. The widespread uses of antibiotics, together with the length of time over which they have been available, have led to the emergence of resistant bacterial pathogens contributing to morbidity and mortality. This study was aimed to assess bacterial isolates and their drug susceptibility patterns from inpatients and outpatients with pus and/or wound discharge. Methods. A cross-sectional study was conducted at the University of Gondar Referral Hospital from March to May, 2014. Wound swab samples were collected from each study participant and inoculated into appropriate media. The bacterial pathogens were identified using standard microbiological methods. Antimicrobial susceptibility tests were performed using disk diffusion technique following Kirby-Bauer method. Results. A total of 137 study subjects were included in the study with bacterial isolation rate of 115 (83.9%. Of all, 81 (59.1% were males. Seventy-seven (57% of the isolates were Gram-negative and 59 (43% were Gram-positive. From the total isolates, Staphylococcus aureus was the most predominant isolate 39/115 (34% followed by Klebsiella species (13%, coagulase negative staphylococci spp. (12% and Pseudomonas aeruginosa. Gram-positive isolates were resistant to ampicillin (86.4%, amoxicillin (83%, penicillin (81.3%, oxacillin (74.6%, and tetracycline (59.4%, while Gram-negative isolates were resistant to amoxicillin (97.4%, ampicillin (94.8%, tetracycline (72.7%, trimethoprim/sulfamethoxazole (66%, and chloramphenicol (54.5%. Conclusion. High prevalence of bacterial isolates was found, Staphylococcus aureus being the most dominant. High rates of multiple drug resistance pathogens to the commonly used antimicrobial agents were isolated. Therefore, concerned bodies should properly monitor the choice of antibiotics to be used as prophylaxis and empiric treatment in the study area.

Prevalence of coagulase-positive staphylococci, other than Staphylococcus aureus, in bovine mastitis.

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Roberson, J R; Fox, L K; Hancock, D D; Gay, J M; Besser, T E

1996-01-01

To determine prevalence and relevance of coagulase-positive Staphylococcus hyicus and S intermedius intramammary infections (IMI) in dairy cows and determine the ability of the 4-hour tube coagulase (TC) test to differentiate the coagulase-positive staphylococci (CPS). Prevalence of CPS was determined for primiparous cows (point prevalence and prevalence at first parturition) and multiparous cows (point prevalence) of 2 herd groups: 10% CPS IMI prevalence = high prevalence (HP). For prevalence, cows of 22 dairy herds. For TC, 1,038 CPS strains isolated from cow milk. Speciation of CPS from aseptically collected composite milk samples was performed. Coagulase-positive isolates from 4 cow groups were tested for their ability to coagulate rabbit plasma by 4 hours: LP and HP primiparous cows at parturition, and LP and HP cows any time after first parturition. Of 487 CPS in the prevalence study, 82.1% were S aureus, 17.7% were coagulase-positive S hyicus, and 0.2% were S intermedius. Of all CPS IMI in LP herds, 34% were coagulase-positive S hyicus; of all CPS IMI in HP herds, 9% were coagulase-positive S hyicus. Coagulase-positive S hyicus appeared to persist to the end of lactation in 4 cows (mean linear somatic cell count = 3.7). The TC test was > or = 97% sensitive, < or = 33% specific, and had a predictive value positive range of 60 to 97% for S aureus isolates. Coagulase-positive S hyicus appears capable of inducing chronic, low-grade IMI. Staphylococcus intermedius does not appear to be an important mastitis pathogen. The TC test is not valid to use as the sole method to differentiate CPS species.

Molecular epidemiology of clinical and carrier strains of methicillin resistant Staphylococcus aureus (MRSA in the hospital settings of north India

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Dar Mohammad J

2006-09-01

Full Text Available Abstract Background The study was conducted between 2000 and 2003 on 750 human subjects, yielding 850 strains of staphylococci from clinical specimens (575, nasal cultures of hospitalized patients (100 and eye & nasal sources of hospital workers (50 & 125 respectively in order to determine their epidemiology, acquisition and dissemination of resistance genes. Methods Organisms from clinical samples were isolated, cultured and identified as per the standard routine procedures. Susceptibility was measured by the agar diffusion method, as recommended by the Nat ional Committee for Clinical Laboratory Standards (NCCLS. The modified method of Birnboin and Takahashi was used for isolation of plasmids from staphylococci. Pulsed-field gel electrophoresis (PFGE typing of clinical and carrier Methicillin resistant Staphylococcus aureus (MRSA strains isolated during our study was performed as described previously. Results It was shown that 35.1% of Staphylococcus aureus and 22.5% of coagulase-negative staphylococcal isolates were resistant to methicillin. Highest percentage of MRSA (35.5% was found in pus specimens (n = 151. The multiple drug resistance of all MRSA (n = 180 and Methicillin resistant Coagulase-negative Staphylococcus aureus (MRCNS (n = 76 isolates was detected. In case of both methicillin-resistant as well as methicillin-sensitive Saphylococcal isolates zero resistance was found to vancomycin where as highest resistance was found to penicillin G followed by ampicillin. It was shown that the major reservoir of methicillin resistant staphylococci in hospitals are colonized/infected inpatients and colonized hospital workers, with carriers at risk for developing endogenous infection or transmitting infection to health care workers and patients. The results were confirmed by molecular typing using PFGE by SmaI-digestion. It was shown that the resistant markers G and T got transferred from clinical S. aureus (JS-105 to carrier S. aureus (JN-49

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

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...... of these species. Approximately half of the mastitis cases were clinical, and in the majority clinical signs were mild. The severity and persistence of intramammary infection were unaffected by CNS species. Fifty-nine percent of the quarter cases were treated with antimicrobials, and the rest were left without...... 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...

Frequency in-vitro antibiotic susceptibility pattern of aerobic isolated from PUS at IIMCT-Railway Hospital Rawalpindi

International Nuclear Information System (INIS)

Khan, A. B.; Hassan, M. U.; Rehman, M. U.; Muzaffar, M.

2006-01-01

A total of 302 samples of pus/pus swabs were cultured aerobically on routine media. One hundred and seventy two bacteria isolated from the samples showing positive growth were identified by standard methods and their antibiotic susceptibility pattern was determined using Kirby-Bauer disk diffusion method. Out of 302 processed samples, 162 samples showed positive result on culture revealing the growth of 172 microorganism of different genera. The spectrum of these isolated bacteria included staphylococcus aureus (51.11%), Escherichia coli (22.9%) pseudomonas aeruginosa (20.93%) and miscellaneous gram negative bacilli (5.81%). The staphylococcus aureus in our study revealed relatively good susceptibility to cloxacillin, flucloxacillin and first generation cephalosporins. In this study 9% of the S. aureus were methicillin resistant. Susceptibility to ampicillin, erythromycin and co-trimoxazole was low. Aminoglycosides and quinolones also showed reasonably good activity against staphylococci. Against Escherichia coli and pseudomonas aeruginosa the activity of quinolones was relatively low when compared with amikacin. Piperacillin+ tazobactam and impepenem/ meropenem revealed a better activity. (author)

Oxacilin-resistant Coagulase-negative staphylococci (CoNS bacteremia in a general hospital at São Paulo city, Brasil Bacteremias por Staphylococcus coagulase negativos oxacilina resistentes em um hospital na cidade de São Paulo, Brasil

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P.A. D'Azevedo

2008-12-01

Full Text Available 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.Staphylococcus coagulase negativos (SCoN, especialmente Staphylococcus epidermidis tem se tornado causa importante de infecções da corrente circulatória nas últimas décadas. Além disso, percentuais de resistência a meticilina entre os SCoN têm aumentado significativamente, levando ao uso de glicopeptídeos nestes pacientes. O objetivo deste estudo foi avaliar onze casos consecutivos de bacteremia clinicamente relevantes por SCoN oxacilina resistentes em um hospital localizado na cidade de São Paulo, Brasil. Cinco diferentes espécies foram identificadas por diferentes métodos fenotípicos, incluindo S. epidermidis (5, S. haemolyticus (3, S. hominis (1, S. warneri (1 e S. cohnii subsp urealyticus (1. Diferentes perfis

The First Report of Drug Resistant Bacteria Isolated from the Brown-Banded Cockroach, Supella longipalpa, in Ahvaz, South-western Iran.

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Babak Vazirianzadeh

2014-06-01

Full Text Available The brown-banded cockroach, Supella longipalpa is known as a carrier of pathogenic bacteria in urban environments, but its role is not well documented regarding the carriage of antibiotic-resistant pathogenic bacteria in Iran. The aim of this study was to determine the resistance bacteria isolated from the brown-banded cockroach in Ahvaz, south west of Iran.Totally 39 cockroaches were collected from kitchen area of houses and identified. All specimens were cultured to isolate the bacterial agents on blood agar and MacConky agar media. The microorganisms were identified using necessary differential and biochemical tests. Antimicrobial susceptibility tests were performed for isolated organisms by Kirby-Bauer's disk diffusion according to NCLI guideline, using 18 antibiotics.From the 39 collected S. langipalpa, 179 bacterial agents were isolated, 92 of alimentary ducts and 87 of external body surfaces. Isolated bacteria from cockroaches were identified as Enterobacter spp., Klebsiella spp., Citrobacter spp., Escherichia coli, Salmonella spp., Proteus spp., coagulase negative staphylococci, Serratia marcescens, Staphylococcus aureus, and Bacillus species. The pattern resistance rates were determined for gram negative bacilli and gram positive cocci regarding 18 antibiotics.The brown-banded cockroach can be involved in the spread of drug resistant bacteria and increases the possibility of contacting human environment to drug resistant bacteria. Therefore, the potential of removing this insect should be improved. This is the first original report of drug resistant bacteria isolated from the brown-banded cockroach of Iran.

High resistance rate against 15 different antibiotics in aerobic gram-negative bacteria isolates of cardiology intensive care unit patients

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Küçükates E

2002-01-01

Full Text Available Aerobic gram negative bacteria were isolated and examined microbiologically from various clinical samples of 602 patients hospitalized between January 1997 and December 2000 in surgical and coronary intensive care units (ICUs. A total of 827 isolates were obtained from 602 patients. The majority of microorganisms were isolated from the respiratory tract (50.3% and blood (39.9%. Pseudomonas spp. were the most frequently isolated gram negative species (32.7%, followed by Acinetobacter spp. (24.0% and Klebsiella pneumoniae (19.4%. High resistance rates to all antibiotics studied were observed. Imipenem and meropenem were the most effective antibiotics against gram negatives.

CD14-negative isolation enhances chondrogenesis in synovial fibroblasts.

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Bilgen, Bahar; Ren, Yuexin; Pei, Ming; Aaron, Roy K; Ciombor, Deborah McK

2009-11-01

Synovial membrane has been shown to contain mesenchymal stem cells. We hypothesized that an enriched population of synovial fibroblasts would undergo chondrogenic differentiation and secrete cartilage extracellular matrix to a greater extent than would a mixed synovial cell population (MSCP). The optimum doses of transforming growth factor beta 1 (TGF-beta1) and insulin-like growth factor 1 (IGF-1) for chondrogenesis were investigated. CD14-negative isolation was used to obtain a porcine cell population enriched in type-B synovial fibroblasts (SFB) from an MSCP. The positive cell surface markers in SFB were CD90, CD44, and cadherin-11. SFB and MSCP were cultured in the presence of 20 ng/mL TGF-beta1 for 7 days, and SFB were demonstrated to have higher chondrogenic potential. Further dose-response studies were carried out using the SFB cells and several doses of TGF-beta1 (2, 10, 20, and 40 ng/mL) and/or IGF-1 (1, 10, 100, and 500 ng/mL) for 14 days. TGF-beta1 supplementation was essential for chondrogenesis and prevention of cell death, whereas IGF-1 did not have a significant effect on the SFB cell number or glycosaminoglycan production. This study demonstrates that the CD14-negative isolation yields an enhanced cell population SFB that is more potent than MSCP as a cell source for cartilage tissue engineering.

Imipenem-resistant Gram-negative bacterial isolates carried by persons upon medical examination in Korea.

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Kim, So Yeon; Shin, Sang Yop; Rhee, Ji-Young; Ko, Kwan Soo

2017-08-01

Carbapenem-resistant Gram-negative bacteria (CR-GNB) have emerged and disseminated worldwide, become a great concern worldwide including Korea. The prevalence of fecal carriage of imipenem-resistant Gram-negative bacteria (IR-GNB) in persons in Korea was investigated. Stool samples were collected from 300 persons upon medical examination. Samples were screened for IR-GNB by using MacConkey agar with 2 μl/ml imipenem. Species were identified by 16S rRNA gene sequence analysis, and antimicrobial susceptibility was determined by the broth microdilution method. In total, 82 IR-GNB bacterial isolates were obtained from 79 (26.3%) out of 300 healthy persons. Multilocus sequence typing analysis showed very high diversity among IR P. aeruginosa, S. maltophilia, and E. cloacae isolates, and pulsed-field gel electrophoresis revealed five main pulsotypes of IR P. mirabilis. As for the presence of metallo-β-lactamases (MBLs), only one IMP-25-producing S. marcescens isolate was identified. Although only one carbapenemase-producing isolate was identified, the high colonization rates with IR-GNB isolates in this study is notable because carriers may be a reservoir for the dissemination of resistant pathogens within the community as well as in health care institutions.

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.

Prevalence of microorganisms associated with udder infections in dairy goats on small-scale farms in Kenya : short communication

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E.N. Ndegwa

2001-07-01

Full Text Available Six hundred and thirty clinically-normal milk samples from dairy goat flocks comprising a mixed population of German Alpine, Toggenburg, Saanen and Galla crosses were exam-ined over a 3-month period to determine the prevalence of bacterial organisms. Bacteria were isolated in 28.7 % of the milk samples (181/630 either singly (92.8 % or in combination (7.2 %. The most prevalent bacterial organisms were Staphylococcus spp. (60.3 %, followed by Micrococcus spp. (17.7 %, Acinetobacter spp. (5 %, Actinomyces spp. (5% and Streptococcus spp. (1.1 %. The Staphylococcus spp. were mainly coagulase negative (64.3 %. Coagulase-- negative staphylococci and coagulase-positive staphylococci accounted for 37.5 % and 22.7 % respectively of the total bacteria isolated. The isolation of bacteria, some of which are important in clinical and subclinical mastitis, in apparently normal caprine milk, indicates that particular attention should be given to the management of these dairy goat flocks in order to avoid the development of cases of clinical mastitis.

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

International Nuclear Information System (INIS)

Varghese, S.; Jose, S.; Varghese, S.; Kuriakose, S.; Jose, S.

2013-01-01

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

Competitive time- and density-dependent adhesion of staphylococci and osteoblasts on crosslinked poly(ethylene glycol)-based polymer coatings in co-culture flow chambers.

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Saldarriaga Fernández, Isabel C; Busscher, Henk J; Metzger, Steve W; Grainger, David W; van der Mei, Henny C

2011-02-01

Biomaterial-associated infections (BAI) remain a serious clinical complication, often arising from an inability of host tissue-implant integration to out-compete bacterial adhesion and growth. A commercial polymer coating based on polyethylene glycol (PEG), available in both chemically inert and NHS-activated forms (OptiChem(®)), was compared for simultaneous growth of staphylococci and osteoblasts. In the absence of staphylococci, osteoblasts adhered and proliferated well on glass controls and on the NHS-reactive PEG-based coating over 48 h, but not on the inert PEG coating. Staphylococcal growth was low on both PEG-based coatings. When staphylococci were pre-adhered on surfaces for 1.5 h to mimic peri-operative contamination, osteoblast growth and spreading was reduced on glass but virtually absent on both reactive and inert PEG-based coatings. Thus although NHS-reactive, PEG-based coatings stimulated tissue-cell interactions in the absence of contaminating staphylococci, the presence of adhering staphylococci eliminated osteoblast adhesion advantages on the PEG surface. This study demonstrates the importance of using bacterial and cellular co-cultures compared to monocultures when assessing functionalized biomaterials coatings for infectious potential. Copyright © 2010 Elsevier Ltd. All rights reserved.

Isolated perifacial lymph node metastasis in oral squamous cell carcinoma with clinically node-negative neck.

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Agarwal, Sangeet Kumar; Arora, Sowrabh Kumar; Kumar, Gopal; Sarin, Deepak

2016-10-01

The incidence of occult perifacial nodal disease in oral cavity squamous cell carcinoma is not well reported. The purpose of this study was to evaluate the incidence of isolated perifacial lymph node metastasis in patients with oral squamous cell carcinoma with a clinically node-negative neck. The study will shed light on current controversies and will provide valuable clinical and pathological information in the practice of routine comprehensive removal of these lymph node pads in selective neck dissection in the node-negative neck. Prospective analysis. This study was started in August 2011 when intraoperatively we routinely separated the lymph node levels from the main specimen for evaluation of the metastatic rate to different lymph node levels in 231 patients of oral squamous cell cancer with a clinically node-negative neck. The current study demonstrated that 19 (8.22%) out of 231 patients showed ipsilateral isolated perifacial lymph node involvement. The incidence of isolated perifacial nodes did not differ significantly between the oral tongue (7.14%) and buccal mucosa (7.75%). Incidence was statistically significant in cases with lower age group (oral squamous cell carcinoma with a clinically node-negative neck. The incidence of isolated perifacial involvement is high in cases of buccal mucosal and tongue cancers. A meticulous dissection of the perifacial nodes seems prudent when treating the neck in oral cavity squamous cell carcinoma. 4 Laryngoscope, 126:2252-2256, 2016. © 2016 The American Laryngological, Rhinological and Otological Society, Inc.

Characterization of fatty acid modifying enzyme activity in staphylococcal mastitis isolates and other bacteria

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Lu Thea

2012-06-01

Full Text Available Abstract Background Fatty acid modifying enzyme (FAME has been shown to modify free fatty acids to alleviate their bactericidal effect by esterifying fatty acids to cholesterol or alcohols. Although it has been shown in previous studies that FAME is required for Staphylococcus aureus survival in skin abscesses, FAME is poorly studied compared to other virulence factors. FAME activity had also been detected in coagulase-negative staphylococci (CNS. However, FAME activity was only surveyed after a bacterial culture was grown for 24 h. Therefore if FAME activity was earlier in the growth phase, it would not have been detected by the assay and those strains would have been labeled as FAME negative. Results Fifty CNS bovine mastitis isolates and several S. aureus, Escherichia coli, and Streptococcus uberis strains were assayed for FAME activity over 24 h. FAME activity was detected in 54% of CNS and 80% S. aureus strains surveyed but none in E. coli or S. uberis. While some CNS strains produced FAME activity comparable to the lab strain of S. aureus, the pattern of FAME activity varied among strains and across species of staphylococci. All CNS that produced FAME activity also exhibited lipase activity. Lipase activity relative to colony forming units of these CNS decreased over the 24 h growth period. No relationship was observed between somatic cell count in the milk and FAME activity in CNS. Conclusions Some staphylococcal species surveyed produced FAME activity, but E. coli and S. uberis strains did not. All FAME producing CNS exhibited lipase activity which may indicate that both these enzymes work in concert to alter fatty acids in the bacterial environment.

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

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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 genes...... showed the monophyly of the strains characterized and the reference strains of P. multocida. The sucrose-negative strains formed two groups, one related to reference strains of P. multocida and the other related to a separate species-like group (taxon 45 of Bisgaard). DNA-DNA hybridization further...... 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-negative...

Plasmids of Staphylococcus cohnii isolated from the intensive-care unit.

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Szewczyk, E M; Rózalska, M; Cieślikowski, T; Nowak, T

2004-01-01

Numerous isolates of both subspecies of Staphylococcus cohnii were found in the environment of the intensive-care unit of a pediatric hospital. These isolates carried in their cells many plasmids, up to fourteen, of a wide range of sizes ( 56 kb). Striking was the occurrence of large plasmids not very common in staphylococci. These were present in > 80% of S. cohnii isolates. Fifty-two different plasmid profiles were found in 79 investigated isolates belonging to S. cohnii ssp. cohnii and S. cohnii ssp. urealyticus. Isolates similar in plasmid profiles were grouped in antibiotic-resistance clusters established for 9 antibiotics (gentamicin, ciprofloxacin, clindamycin, erythromycin, tetracycline, chloramphenicol, mupirocin, trimethoprim-sulfamethoxazole, vancomycin) using the method of unweighted pair group mathematical averages (UPGMA). Many isolates were multiresistant to antibiotics and produced bacteriocins.

Identification of Staphylococcus spp. isolated during the ripening process of a traditional minas cheese

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B.M. Borelli

2011-04-01

Full Text Available The population dynamics of Staphylococcus spp. was studied during the ripening of Canastra Minas cheese at three farms located in the State of Minas Gerais, Brazil. The presence of coagulase (coa, thermonuclease (nuc, and enterotoxin (sea, seb, sec, and sed genes was investigated in Staphylococcus strains isolated during the 60-day cheese-ripening period. The presence of the staphylococcal enterotoxins A, C, and D was also investigated in the cheese samples. Cheese samples that were matured for 0, 7, 15, 30, and 45 days presented staphylococci counts from 10³ to 10(8cfu/g. All isolates considered coagulase-positive by physiological tests had the coa gene. However, no association was observed between the results obtained with biochemical tests and those obtained by PCR using gene-specific primers for coagulase-negative strains. Coagulase and thermonuclease genes occurred simultaneously in 41.3% of Staphylococcus spp. tested. None of the investigated Staphylococcus strains expressed enterotoxins SEA, SEB, SEC, and SED. Enterotoxins A, C, and D were not detected in any of the cheese samples.

Antibiotic resistance, ability to form biofilm and susceptibility to copper alloys of selected staphylococcal strains isolated from touch surfaces in Polish hospital wards

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Anna Różańska

2017-08-01

Full Text Available Abstract Background Despite the employment of sanitary regimes, contact transmission of the aetiological agents of hospital infections is still exceedingly common. The issue of microbe transmission becomes particularly important when facing multidrug-resistant microorganisms such as methicillin-resistant staphylococci. In the case of deficiencies in cleaning and disinfection procedures, hospital equipment made of copper alloys can play an important role, complementing traditional hospital hygiene procedures. The objective of this study was to characterize staphylococcal strains isolated from touch surfaces in Polish hospital wards in terms of their drug resistance, ability to form biofilm and susceptibility to antimicrobial activity of copper alloys. Methods The materials for the study were 95 staphylococcal strains isolated from touch surfaces in 13 different hospital wards from Małopolska province (the south of Poland. Phenotypic and genotypic antibiotic resistance were checked for erythromycin, clindamycin, gentamycin, ciprofloxacin, trimethoprim/sulfamethoxazole and mupirocin. Biofilm formation ability for the tested strains was checked with the use of culture on Congo red agar. Susceptibility to copper, tin bronze, brass and new silver was tested using a modification of the Japanese standard. Results Over 67% of the analysed staphylococcal strains were methicillin-resistant (MR. Four strains were resistant to all of the tested antibiotics, and 14 were resistant to all except mupirocin. Strains classified as MR had significantly increased resistance to the remaining antibiotic groups. About one-third of the analysed strains revealed biofilm-forming ability. Among the majority of species, biofilm-forming and non-biofilm-forming strains were distributed evenly; in the case of S. haemolyticus only, negative strains accounted for 92.8%. Susceptibility to copper alloys was different between strains and rather lower than in the case of the SA

Disinfectant and antibiotic activities: a comparative analysis in Brazilian hospital bacterial isolates

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Guimarães Márcia Aparecida

2000-01-01

Full Text Available Nosocomial infections are an important cause of morbidity and mortality all over the world. It has been shown that appropriate environmental hygienic and disinfection practices can be very helpful to hospital infection control. The purpose of this study was to evaluate the bactericidal activity of some disinfectants against antibiotic-susceptible and antibiotic-resistant hospital bacterial isolates. The susceptibility of 27 clinical isolates to disinfectants and antibiotics was determined by the Association of Official Analytical Chemist?s (AOAC Use-Dilution method and by the Kirby-Bauer method, respectively. All strains tested were susceptible to sodium hypochlorite, glutaraldehyde and to the association quaternary ammonium - formaldehyde - ethyl alcohol disinfectants. However, the susceptibility of strains to phenol and to one quaternary ammonium compound was variable. Among twenty-one antibiotic-multiresistant strains (methicillin-resistant staphylococci, Enterococcus spp, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus mirabilis, Enterobacter cloacae, Serratia marcescens and Escherichia coli eleven (52% and eight (38% strains were resistant to the quaternary ammonium and phenol compounds, respectively. Among six isolates that demonstrated susceptibility to antibiotics (staphylococci, Enterococcus spp, P. mirabilis, E. cloacae and E. coli two strains (33% showed resistance to these disinfectants. The results demonstrated the lack of correlation between antibiotic-susceptibility and susceptibility to disinfectants in hospital strains.

Different phenotypic and molecular mechanisms associated with multidrug resistance in Gram-negative clinical isolates from Egypt

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Helmy OM

2017-12-01

Full Text Available Omneya M Helmy, Mona T Kashef Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo, Egypt Objectives: We set out to investigate the prevalence, different mechanisms, and clonal relatedness of multidrug resistance (MDR among third-generation cephalosporin-resistant Gram-negative clinical isolates from Egypt.Materials and methods: A total of 118 third-generation cephalosporin-resistant Gram-negative clinical isolates were included in this study. Their antimicrobial susceptibility pattern was determined using Kirby–Bauer disk diffusion method. Efflux pump-mediated resistance was tested by the efflux-pump inhibitor-based microplate assay using chlorpromazine. Detection of different aminoglycoside-, β-lactam-, and quinolone-resistance genes was done using polymerase chain reaction. The genetic diversity of MDR isolates was investigated using random amplification of polymorphic DNA.Results: Most of the tested isolates exhibited MDR phenotypes (84.75%. The occurrence of efflux pump-mediated resistance in the different MDR species tested was 40%–66%. Acinetobacter baumannii isolates showed resistance to most of the tested antibiotics, including imipenem. The blaOXA-23-like gene was detected in 69% of the MDR A. baumannii isolates. The MDR phenotype was detected in 65% of Pseudomonas aeruginosa isolates, of which only 23% exhibited efflux pump-mediated resistance. On the contrary, efflux-mediated resistance to piperacillin and gentamicin was recorded in 47.5% of piperacillin-resistant and 25% of gentamicin-resistant MDR Enterobacteriaceae. Moreover, the plasmid-mediated quinolone-resistance genes (aac(6’-Ib-cr, qnrB, and qnrS were detected in 57.6% and 83.33% of quinolone-resistant MDR Escherichia coli and Klebsiella pneumoniae isolates, respectively. The β-lactamase-resistance gene blaSHV-31 was detected for the first time in one MDR K. pneumoniae isolate from an endotracheal tube specimen in Egypt

The frequency of resistance to antibiotics of most frequently isolated bacteria from blood cultures during the period 1997-2002

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Mirović Veljko

2004-01-01

Full Text Available The aim of this study was to determine the frequency of resistance to antibiotics of the most frequently isolated bacteria from blood cultures of hospitalized patients during the period 1997-2002. The resistance to antibiotics was determined by disk diffusion method according to National Committee for Clinical Laboratory Standards procedures. The majority of staphylococci isolates were resistant to methicillin, and the proportion of methicillin-resistant Staphylococcus aureus was stable (76.8-81.6%, during the follow-up period. None of the staphylococci isolates were resistant to vancomycin, but there was a very high incidence of high-level resistance of enterococci to aminoglycosides (47.2-72.2%. In 1998, only one strain among enterococci was resistant to vancomycin (Enterococcus faecium, VanA fenotype. Enterococcus spp isolates expressed variable frequency of resistance to ampicillin (15-40.1% during the follow-up period. Among Enterobacteriaceae there were no isolates resistant to imipenem, but dramatic increase of the resistance to ceftriaxone was found from 35.9% in 1997 to 95.9% in 2002 (p<0.001. Extended spectrum beta-lactamases production was found in all the species of enterobacteria isolates. Resistance to imipenem was observed in Acinetobacter spp isolates in 2002 for the first time. Pseudomonas spp isolates expressed high and very variable resistance to all antibiotics tested during the follow-up period.

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.

Science.gov (United States)

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.

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.

Antibiotic resistance in bacteria Staphylococcus spp. isolated from samples of raw sheep's milk

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Milan Vasiľ

2016-12-01

Full Text Available From samples of raw sheep's milk were determined results of bacteriological examination from two herds in region of Eastern Slovakia in three years lasting study. The occurrence of Staphylococcus spp. 41.6% (124 was determined from 298 samples. The seven species of staphylococci were on a regular basis isolated: S. epidermidis (34, S. chromogenes (26, S. aureus (16. Alternately have been recorded S. warneri (16, S. schleiferi (15, S. haemolyticus (9 and S. xylosus (8. All isolated pathogens were tested by in vitro test on Mueller-Hinton agar by disc methods on resistance to 10 types of antibiotics.  Highest value of resistance was determined to Penicilin 21.0%, Neomycin 10.5% and Novobiocin 9.7%. Lower resistance was in to Oxacilin 7.2% and Amoxicilin 6.5%. Minimal resistance was founded to Cefoxitin 0.8%, Linkomycin 2.4%, Erytromycin, and Streptomycin 3.2%. Was founded total resistance (21.0% to all antibiotics in S. epidermidis (34 during the three years, S. chromogenes (26 showed resistance to 8 types of antibiotics (12.9%, S. aureus (16 to 6 antibiotics (10.5% and S. warneri (16 to 4 antibiotics (5.6%. It was confirmed that sheep's milk remains a major source of staphylococci. Bacteria in comparison with isolates from cows' raw milk, showed lower values of resistance, but were resistant to more than two antibiotics. Recorded occurrence of resistance in staphylococci may be connected with a minimum use of antibiotics in the treatment of mastitis and other diseases in sheep herds. Reported resistance to the tested antibiotics became the basis for the recommendation to use preparations to treat mastitis in sheep principally by the detection of resistance to antibiotics contained.

Incidence of bovine clinical mastitis in Jammu region and antibiogram of isolated pathogens

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Adil Majid Bhat

2017-08-01

Full Text Available Aim: This study was conducted to evaluate the incidence of clinical mastitis in bovines of Jammu region, to identify the infectious organisms responsible for it, and the antimicrobial sensitivity of isolated pathogens. Materials and Methods: The study was conducted on cases that were presented to the Medicine Division of Teaching Veterinary Clinical Complex, Faculty of Veterinary Sciences and Animal Husbandry, R.S. Pura, Jammu, Jammu and Kashmir. A total of 260 cases of bovines were presented from June 30, 2012, to July 01, 2013, out of which 30 cases were of clinical mastitis. The diagnosis of clinical mastitis was made on the basis of history and clinical examination of affected animals. Results: Animal and quarter-wise incidence of clinical mastitis were found to be 11.5% and 5.76%, respectively. Of the 23 isolates obtained, Staphylococcus aureus (60.87% was the most frequently isolated organism, followed by coagulase negative Staphylococci (13.04%, Streptococcus uberis (4.35%, Streptococcus dysgalactiae (8.69%, and Escherichia coli (13.04%. The antimicrobial sensitivity of isolates revealed maximum sensitivity to enrofloxacin, gentamicin, amoxicillin/ sulbactam, ceftriaxone/tazobactam, ceftizoxime, ampicillin/sulbactam and least sensitivity for oxytetracycline and penicillin. Conclusion: Staphylococcus spp. is the major causative agent of clinical mastitis in bovines of Jammu region. The causative agents of the clinical mastitis were most sensitive to enrofloxacin and gentamicin.

The Antibiotic Resistance Profiles of Bacterial Strains Isolated from Patients with Hospital-Acquired Bloodstream and Urinary Tract Infections

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Hamed Ghadiri

2012-01-01

Full Text Available Treatment of nosocomial infections is becoming difficult due to the increasing trend of antibiotics resistance. Current knowledge on antibiotic resistance pattern is essential for appropriate therapy. We aimed to evaluate antibiotic resistance profiles in nosocomial bloodstream and urinary tract pathogens. A total of 129 blood stream and 300 urinary tract positive samples were obtained from patients referring to Besat hospital over a two-year period (2009 and 2010. Antibiotic sensitivity was ascertained using the Kirby-Bauer disk diffusion technique according to CLSI guidelines. Patient's data such as gender and age were recorded. The ratio of gram-negative to gram-positive bacteria in BSIs was 1.6 : 1. The most prevalent BSI pathogen was Coagulase-Negative Staphylococci (CoNS. The highest resistance rate of CoNS was against penicillin (91.1% followed by ampicillin (75.6%, and the lowest rate was against vancomycin (4.4%. Escherichia coli was the most prevalent pathogen isolated from urinary tract infections (UTIs. Ratio of gram-negative to gram-positive bacteria was 3.2 : 1. The highest resistance rate of E. coli isolates was against nalidixic acid (57.7%. The present study showed that CoNS and E. coli are the most common causative agents of nosocomial BSIs and UTIs, and control of infection needs to be addressed in both antibiotic prescription and general hygiene.

Mastite subclínica por Staphylococcus coagulase negativa em ovinos de corte

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T.S. Tejada

2012-08-01

Full Text Available Microorganisms that cause mastitis were isolated in a herd of sheep and their sensitivity to antimicrobials was tested. Twenty-one sheep had their mammary glands monitored during lactation. Eighty-eight milk sample from CMT-positive sheep were analyzed in order to isolate the etiologic agent of mastitis. Coagulase negative Staphylococcus were isolated from 29.5% of samples. Ampicillin and penicillin were the antimicrobials with higher number of resistant strains. The importance of coagulase negative staphylococci has been demonstrated, especially S. lentus and S. sciuri as a cause of ovine mastitis.

Intra-species diversity and epidemiology varies among coagulase-negative Staphylococcus species causing bovine intramammary infections.

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Piessens, V; De Vliegher, S; Verbist, B; Braem, G; Van Nuffel, A; De Vuyst, L; Heyndrickx, M; Van Coillie, E

2012-02-24

Although many studies report coagulase-negative staphylococci (CNS) as the predominant cause of subclinical bovine mastitis, their epidemiology is poorly understood. In the current study, the genetic diversity within four CNS species frequently associated with bovine intramammary infections, Staphylococcus haemolyticus, S. simulans, S. chromogenes, and S. epidermidis, was determined. For epidemiological purposes, CNS genotypes recovered from bovine milk collected on six Flemish dairy farms were compared with those from the farm environment, and their distribution within the farms was investigated. Genetic diversity was assessed by two molecular typing techniques, amplification fragment length polymorphism (AFLP) and random amplification of polymorphic DNA (RAPD) analysis. Subtyping revealed the highest genetic heterogeneity among S. haemolyticus isolates. A large variety of genotypes was found among environmental isolates, of which several could be linked with intramammary infection, indicating that the environment could act as a potential source for infection. For S. simulans, various genotypes were found in the environment, but a link with IMI was less obvious. For S. epidermidis and S. chromogenes, genetic heterogeneity was limited and the sporadic isolates from environment displayed largely the same genotypes as those from milk. The higher clonality of the S. epidermidis and S. chromogenes isolates from milk suggests that specific genotypes probably disseminate within herds and are more udder-adapted. Environmental sources and cow-to-cow transmission both seem to be involved in the epidemiology of CNS, although their relative importance might substantially vary between species. Copyright © 2011 Elsevier B.V. All rights reserved.

Antimicrobial activity of extracts from macroalgae Ulva lactuca against clinically important Staphylococci is impacted by lunar phase of macroalgae harvest.

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Deveau, A M; Miller-Hope, Z; Lloyd, E; Williams, B S; Bolduc, C; Meader, J M; Weiss, F; Burkholder, K M

2016-05-01

Staphylococcus aureus is a common human bacterial pathogen that causes skin and soft tissue infections. Methicillin-resistant Staph. aureus (MRSA) are increasingly drug-resistant, and thus there is great need for new therapeutics to treat Staph. aureus infections. Attention has focused on potential utility of natural products, such as extracts of marine macroalgae, as a source of novel antimicrobial compounds. The green macroalgae Ulva lactuca produces compounds inhibitory to human pathogens, although the effectiveness of U. lactuca extracts against clinically relevant strains of Staph. aureus is poorly understood. In addition, macroalgae produce secondary metabolites that may be influenced by exogenous factors including lunar phase, but whether lunar phase affects U. lactuca antimicrobial capacity is unknown. We sought to evaluate the antibacterial properties of U. lactuca extracts against medically important Staphylococci, and to determine the effect of lunar phase on antimicrobial activity. We report that U. lactuca methanolic extracts inhibit a range of Staphylococci, and that lunar phase of macrolagae harvest significantly impacts antimicrobial activity, suggesting that antimicrobial properties can be maximized by manipulating time of algal harvest. These findings provide useful parameters for future studies aimed at isolating and characterizing U. lactuca anti-Staphylococcal agents. The growing prevalence of antibiotic-resistant human pathogens such as methicillin-resistant Staphylococcus aureus (MRSA) has intensified efforts towards discovery and development of novel therapeutics. Marine macroalgae like Ulva lactuca are increasingly recognized as potential sources of antimicrobials, but the efficacy of U. lactuca extracts against common, virulent strains of Staph. aureus is poorly understood. We demonstrate that U. lactuca methanolic extracts inhibit a variety of clinically relevant Staphylococcus strains, and that the antimicrobial activity can

Microbiological and molecular characterization of Staphylococcus hominis isolates from blood.

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Soraya Mendoza-Olazarán

Full Text Available BACKGROUND: Among Coagulase-Negative Staphylococci (CoNS, Staphylococcus hominis represents the third most common organism recoverable from the blood of immunocompromised patients. The aim of this study was to characterize biofilm formation, antibiotic resistance, define the SCCmec (Staphylococcal Chromosomal Cassette mec type, and genetic relatedness of clinical S. hominis isolates. METHODOLOGY: S. hominis blood isolates (n = 21 were screened for biofilm formation using crystal violet staining. Methicillin resistance was evaluated using the cefoxitin disk test and the mecA gene was detected by PCR. Antibiotic resistance was determined by the broth microdilution method. Genetic relatedness was determined by pulsed-field gel electrophoresis (PFGE and SCCmec typed by multiplex PCR using two different methodologies described for Staphylococcus aureus. RESULTS: Of the S. hominis isolates screened, 47.6% (10/21 were categorized as strong biofilm producers and 23.8% (5/21 as weak producers. Furthermore, 81% (17/21 of the isolates were methicillin resistant and mecA gene carriers. Resistance to ampicillin, erythromycin, and trimethoprim was observed in >70% of isolates screened. Each isolate showed a different PFGE macrorestriction pattern with similarity ranging between 0-95%. Among mecA-positive isolates, 14 (82% harbored a non-typeable SCCmec type: eight isolates were not positive for any ccr complex; four contained the mec complex A ccrAB1 and ccrC, one isolate contained mec complex A, ccrAB4 and ccrC, and one isolate contained the mec complex A, ccrAB1, ccrAB4, and ccrC. Two isolates harbored the association: mec complex A and ccrAB1. Only one strain was typeable as SCCmec III. CONCLUSIONS: The S. hominis isolates analyzed were variable biofilm producers had a high prevalence of methicillin resistance and resistance to other antibiotics, and high genetic diversity. The results of this study strongly suggested that S. hominis isolates harbor

Methicillin-Resistant Staphylococcus aureus from Brazilian Dairy Farms and Identification of Novel Sequence Types.

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Oliveira, C J B; Tiao, N; de Sousa, F G C; de Moura, J F P; Santos Filho, L; Gebreyes, W A

2016-03-01

The aim of this study was to investigate the phenotypic and genotypic diversity and anti-microbial resistance among staphylococci of dairy herds that originated from Paraiba State, north-eastern Brazil, a region where such studies are rare. Milk samples (n = 552) were collected from 15 dairy farms. Isolates were evaluated for anti-microbial susceptibility by Kirby-Bauer disc diffusion method. Confirmation of methicillin-resistant Staphylococcus aureus (MRSA) was performed using multiplex PCR targeting mecA and nuc genes in addition to phenotypic assay based on PBP-2a latex agglutination. Clonal relatedness of isolates was determined by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) genotyping. Staphylococci were detected in 269 (49%) of the samples. Among these, 65 (24%) were S. aureus. The remaining 204 isolates were either coagulase-negative staphylococci (n = 188; 70%) or coagulase positive other than S. aureus (n = 16; 6%). Staphylococci were cultured in seven (35%) of the 20 hand swab samples, from which five isolates were S. aureus. The isolates were most commonly resistant against penicillin (43%), ampicillin (38%) and oxacillin (27%). The gene mecA was detected in 21 S. aureus from milk and in one isolate from a milker's hand. None of the isolates were resistant to vancomycin. PFGE findings showed high clonal diversity among the isolates. Based on MLST, we identified a total of 11 different sequence types (STs 1, 5, 6, 83, 97, 126, 1583, 1622, 1623, 1624 and 1625) with four novel STs (ST1622-ST1625). The findings show that MRSA is prevalent in milk from semi-extensive dairy cows in north-eastern Brazil, and further investigation on its extent in various types of milk production systems and the farm-to-table continuum is warranted. © 2015 Blackwell Verlag GmbH.

Microbiological and molecular identification of bacterial species isolated from nasal and oropharyngeal mucosa of fuel workers in Riyadh, Saudi Arabia.

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AlWakeel, Suaad S

2017-09-01

This study aimed to determine the bacterial species colonizing the nasal and oropharyngeal mucosa of fuel workers in Central Riyadh, Saudi Arabia on a microbiological and molecular level. Throat and nasal swab samples were obtained from 29 fuel station attendants in the period of time extending from March to May 2014 in Riyadh, Saudi Arabia. Microbiological identification techniques were utilized to identify the bacterial species isolated. Antibiotic sensitivity was assessed for each of the bacterial isolates. Molecular identification techniques based on PCR analysis of specific genomic sequences was conducted and was the basis on which phylogeny representation was done for 10 randomly selected samples of the isolates. Blood was drawn and a complete blood count was conducted to note the hematological indices for each of the study participants. Nineteen bacterial species were isolated from both the nasal cavity and the oropharynx including Streptococcus thoraltensis , alpha-hemolytic streptococci, Staphylococcus hominis , coagulase-negative staphylococci, Leuconostoc mesenteroides , Erysipelothrix rhusiopathiae and several others. We found 100% sensitivity of the isolates to ciprofloxacin, cefuroxime and gentamicin. Whereas cefotaxime and azithromycin posted sensitivities of 85.7% and 91.4%, respectively. Low sensitivities (fuel products may be a contributing factor to bacterial colonization of the respiratory tract in fuel workers.

Comprehensive Phylogenetic Analysis of Bovine Non-aureus Staphylococci Species Based on Whole-Genome Sequencing

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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

In vitro efficacy and pharmacodynamic indices for antibiotics against coagulase-negative staphylococcus endophthalmitis isolates.

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Harper, Tom; Miller, Darlene; Flynn, Harry W

2007-05-01

To compare pharmacodynamic indices and minimal inhibitory concentrations for vancomycin, gatifloxacin, moxifloxacin, linezolid, and combined quinupristin and dalfopristin for historic and current human coagulase-negative staphylococcus (CoNS) endophthalmitis isolates. Experimental study. Fifty-nine CoNS endophthalmitis isolates retrieved from patients at the Bascom Palmer Eye Institute from 1993 through 2006. Coagulase-negative staphylococcal endophthalmitis isolates were recovered from the microbiology specimen bank, rehydrated, and processed for susceptibility testing using standard microbiological protocols. E tests were used to determine and compare mean inhibitory concentration for 50% of isolates (MIC50) and mean inhibitory concentration for 90% of isolates (MIC90) values. Peak concentration (C(max)) was defined as the maximum attainable aqueous concentration using topical or oral therapy, or both. The MIC50 and MIC90 values for each antibiotic are the minimum concentrations that inhibit 50% and 90% of CoNS endophthalmitis isolates, respectively. Significance was determined by the McNemar test. Pharmacodynamic indices (C(max)/MIC) were calculated using determined MIC values and published intraocular drug concentrations for topical and oral dosing. The pharmacodynamic index was defined as the achievable aqueous humor concentration of an antibiotic divided by the concentration of the antibiotic required to inhibit a specified percentage of microbiologic isolates. Pharmacodynamic indices for new and conventional antibiotics. General in vitro susceptibility patterns in descending order were vancomycin (100%), linezolid (100%), quinupristin and dalfopristin (98%), moxifloxacin (48%), and gatifloxacin (47%). The corresponding MIC50 and MIC90 values were vancomycin, 2 microg/ml and 3 microg/ml, respectively; linezolid, 1 microg/ml and 4 microg/ml; quinupristin and dalfopristin, 0.25 microg/ml and 0.5 microg/ml; moxifloxacin, 0.75 microg/ml and > or =32 microg

Trends in udder health and emerging mastitogenic pathogens in South African dairy herds

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I.M. Petzer

2009-05-01

Full Text Available The aim of this study was to retrospectively analyse the results of milk samples obtained from South African dairy herds during the period 1996 to April 2007 in order to identify possible trends in isolates of microorganisms and their pathogenicity under field conditions. Milk samples were obtained from 7 of the 9 provinces in South Africa where there are low numbers of dairy cows. Although there is scientific limitation to a country wide survey, such as the variation in herd size, management skills, parity, milk yield, milking frequency and other parameters, the size of this database helps to give a fair indication of general udder health in South Africa. Cytology and routine bacteriology were performed on 379 000 milk samples of lactating cows and bacteriology on 11 946 samples from non-lactating cows. According to the results obtained, mastitis did not decrease in South Africa over the test period. The prevalence of mastitis and teat canal infection was lowest in 2002. Mastitis and teat canal infection increased from 2002 to 2006 from 8.1 % and 24.1 % to 15.4 and 30.0 % respectively. The percentage of mastitogenic pathogens isolated from cows over these years also varied. Previously unknown or almost eradicated mastitogenic pathogens such as αβ haemolytic Staphylococcus aureus which is thought to be of human origin, Streptococcus agalactiae and Enterococcus canis were responsible for numerous mastitis outbreaks seen in the test samples. Coagulase-negative staphylococci were the most frequently isolated bacteria in milk samples from both lactating and dry cows, followed by Staphylococcus aureus and Streptococcus agalactiae. Although Staphylococcus aureus remained the principal mastitogenic pathogen in South Africa, owing to its chronic nature and resultant economic losses, most cases of mastitis were caused by coagulase-negative staphylococci. This finding increases the importance of coagulase-negative staphylococci (formerly described as a

Comparative activity of ceftobiprole against Gram-positive and Gram-negative isolates from Europe and the Middle East: the CLASS study.

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Rossolini, Gian M; Dryden, Matthew S; Kozlov, Roman S; Quintana, Alvaro; Flamm, Robert K; Läuffer, Jörg M; Lee, Emma; Morrissey, Ian; CLASS Study Group

2011-01-01

to assess the in vitro activity of ceftobiprole and comparators against a recent collection of Gram-positive and Gram-negative pathogens, in order to detect potential changes in susceptibility patterns, and to evaluate the Etest assay for ceftobiprole susceptibility testing. contemporary Gram-positive and Gram-negative isolates (excluding extended-spectrum β-lactamase-producing isolates) from across Europe and the Middle East were collected, and their susceptibility to ceftobiprole, vancomycin, teicoplanin, linezolid, ceftazidime and cefepime was assessed using the Etest method. Quality testing [using Etest and broth microdilution (BMD)] was conducted at a central reference laboratory. some 5041 Gram-positive and 4026 Gram-negative isolates were included. Against Gram-positive isolates overall, ceftobiprole had the lowest MIC50 (0.5 mg/L), compared with 1 mg/L for its comparators (vancomycin, teicoplanin and linezolid). Against methicillin-resistant Staphylococcus aureus, all four agents had a similar MIC90 (2 mg/L), but ceftobiprole had a 4-fold better MIC90 (0.5 mg/L) against methicillin-susceptible strains. Only 38 Gram-positive isolates were confirmed as ceftobiprole resistant. Among Gram-negative strains, 86.9%, 91.7% and 95.2% were susceptible to ceftobiprole, ceftazidime and cefepime, respectively. Pseudomonas aeruginosa was less susceptible to all three antimicrobials than any other Gram-negative pathogen. There was generally good agreement between local Etest results and those obtained at the reference laboratory (for ceftobiprole: 86.8% with Gram-negatives; and 94.7% with Gram-positives), as well as between results obtained by BMD and Etest methods (for ceftobiprole: 98.2% with Gram-negatives; and 98.4% with Gram-positives). ceftobiprole exhibits in vitro activity against a wide range of Gram-positive and Gram-negative pathogens, including multidrug-resistant strains. No changes in its known susceptibility profile were identified.

SOME ASPECTS OF NASAL CARRIAGE OF STAPHYLOCOCCI.

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NOBLE, W C; WILLIAMS, R E; JEVONS, M P; SHOOTER, R A

1964-01-01

The nasal carrier status of 3,736 patients was determined throughout their stay in hospital. The carrier rate on admission, which was highest in patients under 20 years of age, did not appear to vary with season.The carriage of strains resistant to penicillin increased with the patients' stay in hospital from 13.% on admission to 20.5% on discharge, and the acquisition of these strains was enhanced by the administration of antibiotics. Patients discharged from hospital carrying strains of staphylococci acquired in hospital lost them more readily than patients discharged carrying the strain which they had carried on admission, 31% of those discharged carrying strains resistant to penicillin and tetracycline being readmitted carrying these strains compared with 69% of those discharged carrying strains sensitive to these antibiotics.

Atorvastatin affects negatively respiratory function of isolated endothelial mitochondria.

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Broniarek, Izabela; Jarmuszkiewicz, Wieslawa

2018-01-01

The purpose of this research was to elucidate the direct effects of two popular blood cholesterol-lowering drugs used to treat cardiovascular diseases, atorvastatin and pravastatin, on respiratory function, membrane potential, and reactive oxygen species formation in mitochondria isolated from human umbilical vein endothelial cells (EA.hy926 cell line). Hydrophilic pravastatin did not significantly affect endothelial mitochondria function. In contrast, hydrophobic calcium-containing atorvastatin induced a loss of outer mitochondrial membrane integrity, an increase in hydrogen peroxide formation, and reductions in maximal (phosphorylating or uncoupled) respiratory rate, membrane potential and oxidative phosphorylation efficiency. The atorvastatin-induced changes indicate an impairment of mitochondrial function at the level of ATP synthesis and at the level of the respiratory chain, likely at complex I and complex III. The atorvastatin action on endothelial mitochondria was highly dependent on calcium ions and led to a disturbance in mitochondrial calcium homeostasis. Uptake of calcium ions included in atorvastatin molecule induced mitochondrial uncoupling that enhanced the inhibition of the mitochondrial respiratory chain by atorvastatin. Our results indicate that hydrophobic calcium-containing atorvastatin, widely used as anti-atherosclerotic agent, has a direct negative action on isolated endothelial mitochondria. Copyright © 2017. Published by Elsevier Inc.

Antibacterial Effect of Copper on Microorganisms Isolated from Bovine Mastitis

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Reyes-Jara, Angelica; Cordero, Ninoska; Aguirre, Juan; Troncoso, Miriam; Figueroa, Guillermo

2016-01-01

The antimicrobial properties of copper have been recognized for several years; applying these properties to the prevention of diseases such as bovine mastitis is a new area of research. The aim of the present study was to evaluate in vitro the antimicrobial activity of copper on bacteria isolated from subclinical and clinical mastitis milk samples from two regions in Chile. A total of 327 microorganisms were recovered between March and September 2013, with different prevalence by sample origin (25 and 75% from the central and southern regions of Chile, respectively). In the central region, Escherichia coli and coagulase negative Staphylococci (CNS) were the most frequently detected in clinical mastitis cases (33%), while in the southern region S. uberis, S. aureus, and CNS were detected with frequencies of 22, 21, and 18%, respectively. Antibiotic susceptibility studies revealed that 34% of isolates were resistant to one or more antibiotics and the resistance profile was different between bacterial species and origins of isolation of the bacteria. The minimum inhibitory concentration of copper (MIC-Cu) was evaluated in all the isolates; results revealed that a concentration as low as 250 ppm copper was able to inhibit the great majority of microorganisms analyzed (65% of isolates). The remaining isolates showed a MIC-Cu between 375 and 700 ppm copper, and no growth was observed at 1000 ppm. A linear relationship was found between the logarithm of viable bacteria number and time of contact with copper. With the application of the same concentration of copper (250 ppm), CNS showed the highest tolerance to copper, followed by S. uberis and S. aureus; the least resistant was E. coli. Based on these in vitro results, copper preparations could represent a good alternative to dipping solutions, aimed at preventing the presence and multiplication of potentially pathogenic microorganisms involved in bovine mastitis disease. PMID:27199953

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

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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

Aerobic bacteria cultured from the mouth of the American opossum (Didelphis virginiana) with reference to bacteria associated with bite infections.

OpenAIRE

Howell, J M; Dalsey, W C

1990-01-01

The American opossum inflicts bite injuries both when hunted for food and when accidentally provoked when handled in captivity. This study involved aerobically culturing organisms from the mouths of seven wild opossums (Didelphis virginiana). Isolates included streptococci, coagulase-positive and -negative staphylococci, Aeromonas spp., Citrobacter freundii, Eikenella corrodens, and Escherichia coli.

Evidence for the evolutionary steps leading to mecA-mediated β-lactam resistance in staphylococci

DEFF Research Database (Denmark)

Rolo, Joana; Worning, Peder; Boye Nielsen, Jesper

2017-01-01

the most primitive staphylococci. In this study we aimed to identify evolutionary steps linking these mecA precursors to the β-lactam resistance gene mecA and the resistance phenotype. We sequenced genomes of 106 S. sciuri, S. vitulinus and S. fleurettii strains and determined their oxacillin...

Inducible Clindamycin Resistance in Staphylococcus Species

International Nuclear Information System (INIS)

Afridi, F. I.; Zeb, M.; Farooqi, B. J.; Murtaza, G.; Hussain, A.

2014-01-01

Objective: To determine the frequency of inducible clindamycin resistance in clinical isolates of Staphylococcus species by phenotypic D-test. Study Design: Observational study. Place and Duration of Study: Ziauddin University Hospital, Karachi, from July to December 2011. Methodology: Consecutive clinical isolates of Staphylococcus species were collected and identified by conventional microbiological techniques. Antimicrobial susceptibility testing and inducible clindamycin resistance was carried out by performing D-test using CLSI criteria. Methicillin resistance was detected by using Cefoxitin disk as a surrogate marker. Statistical analysis was performed by SPSS version-17. Results: A total of 667 clinical isolates of Staphylococcus species were obtained during the study period. In these isolates, 177 (26.5%) were Staphylococcus aureus, and 490 (73.5%) were coagulase negative Staphylococci. The total frequency of inducible clindamycin resistance among isolates of Staphylococcus species was 120/667 (18%). Frequency of inducible clindamycin resistance among coagulase negative Staphylococci group and Staphylococcus aureus group were 18.57% and 16.38% respectively. Median age of patients in D-test positive group was 19.5 (1 - 54) years. Conclusion: The frequency of inducible clindamycin resistance among Staphylococcus species may differ in different hospital setup. Clinical microbiology laboratories should implement testing simple and effective D-test on all Staphylococcus species. D-test positive isolates should be reported clindamycin resistant to decrease treatment failure. (author)

In vitro antibacterial activity of doripenem against clinical isolates from French teaching hospitals: proposition of zone diameter breakpoints.

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Lascols, C; Legrand, P; Mérens, A; Leclercq, R; Armand-Lefevre, L; Drugeon, H B; Kitzis, M D; Muller-Serieys, C; Reverdy, M E; Roussel-Delvallez, M; Moubareck, C; Lemire, A; Miara, A; Gjoklaj, M; Soussy, C-J

2011-04-01

The aims of the study were to determine the in vitro activity of doripenem, a new carbapenem, against a large number of bacterial pathogens and to propose zone diameter breakpoints for clinical categorization in France according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) minimum inhibitory concentration (MIC) breakpoints. The MICs of doripenem were determined by the broth microdilution method against 1,547 clinical isolates from eight French hospitals. The disk diffusion test was performed (10-μg discs) according to the Comité de l'Antibiogramme de la Société Française de Microbiologie (CASFM) method. The MIC(50/90) (mg/L) values were as follows: methicillin-susceptible Staphylococcus aureus (MSSA) (0.03/0.25), methicillin-resistant Staphylococcus aureus (MRSA) (1/2), methicillin-susceptible coagulase-negative staphylococci (MSCoNS) (0.03/0.12), methicillin-resistant coagulase-negative staphylococci (MRCoNS) (2/8), Streptococcus pneumoniae (0.016/0.25), viridans group streptococci (0.016/2), β-hemolytic streptococci (≤0.008/≤0.008), Enterococcus faecalis (2/4), Enterococcus faecium (128/>128), Enterobacteriaceae (0.06/0.25), Pseudomonas aeruginosa (0.5/8), Acinetobacter baumannii (0.25/2), Haemophilus influenzae (0.12/0.25), and Moraxella catarrhalis (0.03/0.06). According to the regression curve, the zone diameter breakpoints were 24 and 19 mm for MICs of 1 and 4 mg/L, respectively. This study confirms the potent in vitro activity of doripenem against Pseudomonas aeruginosa, Acinetobacter, Enterobacteriaceae, MSSA, MSCoNS, and respiratory pathogens. According to the EUCAST MIC breakpoints (mg/L) ≤1/>4 for Enterobacteriaceae, Pseudomonas aeruginosa, and Acinetobacter, and ≤1/>1 for streptococci, pneumococci, and Haemophilus, the zone diameter breakpoints could be (mm) ≥24/<19 and ≥24/<24, respectively.

A Stewart isolator with high-static-low-dynamic stiffness struts based on negative stiffness magnetic springs

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Zheng, Yisheng; Li, Qingpin; Yan, Bo; Luo, Yajun; Zhang, Xinong

2018-05-01

In order to improve the isolation performance of passive Stewart platforms, the negative stiffness magnetic spring (NSMS) is employed to construct high static low dynamic stiffness (HSLDS) struts. With the NSMS, the resonance frequencies of the platform can be reduced effectively without deteriorating its load bearing capacity. The model of the Stewart isolation platform with HSLDS struts is presented and the stiffness characteristic of its struts is studied firstly. Then the nonlinear dynamic model of the platform including both geometry nonlinearity and stiffness nonlinearity is established; and its simplified dynamic model is derived under the condition of small vibration. The effect of nonlinearity on the isolation performance is also evaluated. Finally, a prototype is built and the isolation performance is tested. Both simulated and experimental results demonstrate that, by using the NSMS, the resonance frequencies of the Stewart isolator are reduced and the isolation performance in all six directions is improved: the isolation frequency band is increased and extended to a lower-frequency level.

Changes of the Quinolones Resistance to Gram-positive Cocci Isolated during the Past 8 Years in the First Bethune Hospital

Science.gov (United States)

Xu, Jiancheng; Chen, Qihui; Yao, Hanxin; Zhou, Qi

This study was to investigate the quinolones resistance to gram-positive cocci isolated in the First Bethune Hospital during the past 8 years. 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). The rates of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative Staphylococci (MRCNS) were 50.8%∼83.3% and 79.4%∼81.5%during the past 8 years, respectively. In recent 8 years, the quinolones resistance to gram-positive cocci had increased. Monitoring of the quinolones resistance to gram-positive cocci should be strengthened. The change of the antimicrobial resistance should be investigated in order to guide rational drug usage in the clinic and prevent bacterial strain of drug resistance from being transmitted.

Production of enterotoxins of Staphylococcus spp. isolated from samples of sheep milk

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František Zigo

2014-02-01

Full Text Available In our study was followed occurrence of mastitis in herd of 430 sheep of breed zoslachtena valaska with hand milking technology examined two times during one lactation season. Individual examination consisted from clinical examination of udder and microbiological examination of milk samples. By PCR was determined presence of genes coding production of enterotoxins, and by ELISA methods production individual types of enterotoxins. From individual forms of mastitis were frequently detected subacute (6.7%, subclinical (5.7% and acute (2.9%. The coagulase-negative staphylococci (CNS were identified in 102 (65.4% from all 156 positive isolates. The CNS and S. aureus caused subacute (5.1%, subclinical (3.9% and acute (2.4% forms of mastitis. The most frequently isolated were S. epidermidis, followed by S. chromogenes and S. xylosus from ewes with subacute and subclinical mastitis. From acute and chronical forms of mastitis were  predominantly isolated S. aureus, S. uberis and S. epidermidis. The production of staphylococcal enterotoxins (SE - SEA, SEB, SEC, SED and the presence of genes sec (3, sea (2, seb (2 and sed (2 were determined in S. aureus, S. epidermidis, S. schleiferi and S. chromogenes, respectively. The results suggested on the high occurrence (12.4% of subacute and subclinical forms. Confirmed production of enterotoxins and presence of genes coding their production present a risk for human health and decreased a quality of milk and products from sheep´s milk.

The in vitro activity of flomoxef compared to four other cephalosporins and imipenem.

Science.gov (United States)

Shah, P M; Knothe, H

1991-01-01

The antibacterial activity of the oxacephalosporin flomoxef was evaluated in comparison to cefpirome, cefuzoname, cefotaxime, ceftazidime, and imipenem against fresh clinical isolates. Flomoxef is an antibiotic with strong antibacterial activity against staphylococci including methicillin-resistant strains and streptococci with the exception of Enterococcus faecalis and Enterococcus faecium. It is very active against gram-negative cocci and rods including gram-positive and gram-negative anaerobes. Against Pseudomonas sp. flomoxef has no activity.

Characterization of Staphylococcus aureus nasal colonization rates ...

African Journals Online (AJOL)

No resistance was detected to linezolid and vancomycin in all isolates. Coagulase negative staphylococci carriage was 87% in all our volunteers and was highest in the HCWs (93%). In conclusion, the study shows the need for a periodic screening of both the community and hospital personnel to adopt strategies for treating ...

Aerobic bacteria cultured from the mouth of the American opossum (Didelphis virginiana) with reference to bacteria associated with bite infections.

Science.gov (United States)

Howell, J M; Dalsey, W C

1990-01-01

The American opossum inflicts bite injuries both when hunted for food and when accidentally provoked when handled in captivity. This study involved aerobically culturing organisms from the mouths of seven wild opossums (Didelphis virginiana). Isolates included streptococci, coagulase-positive and -negative staphylococci, Aeromonas spp., Citrobacter freundii, Eikenella corrodens, and Escherichia coli. PMID:2229365

Characterization of the Extended-Spectrum beta-Lactamase Producers among Non-Fermenting Gram-Negative Bacteria Isolated from Burnt Patients

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Mojdeh Hakemi Vala

2013-09-01

Full Text Available Please cite this article as: Hakemi Vala M, Hallajzadeh M, Fallah F, Hashemi A, Goudarzi H. Characterization of the extended-spectrum beta-lactamase producers among non-fermenting gram-negative bacteria isolated from burnt patients. Arch Hyg Sci 2013;2(1:1-6. Background & Aims of the Study: Extended-spectrum beta-Lactamases (ESBLs represent a major group of beta-lactamases which are responsible for resistance to oxyimino-cephalosporins and aztreonam and currently being identified in large numbers throughout the world. The objective of this study was to characterize ESBL producers among non-fermenter gram-negative bacteria isolated from burnt patients. Materials & Methods: During April to July 2012, 75 non-fermenter gram-negative bacilli were isolated from 240 bacterial cultures collected from wounds of burnt patients admitted to the Burn Unit at Shahid Motahari Hospital (Tehran, Iran. Bacterial isolation and identification was done using standard methods. Antimicrobial susceptibility testing was performed by disk diffusion method for all strains against selected antibiotics and minimum inhibitory concentration was determined by microdilution test. The ability to produce ESBL was detected through double disk synergy test among candidate strains. Results: Of 75 non-fermenter isolates, 47 Pseudomonas aeruginosa and 28 Acinetobacter baumannii were identified. The resistance of P. aeruginosa isolates to tested antibiotics in antibiogram test were 100% to cefpodoxime, 82.98% to ceftriaxone, 78.73% to imipenem, 75% to meropenem, 72.72% to gentamicin, 69.23% to ciprofloxacin and aztreonam, 67.57% to cefepime, 65.95% to ceftazidime, and 61.53% to piperacillin. The results for Acinetobacter baumannii were 100% to ceftazidime, cefepime, ciprofloxacin, imipenem, meropenem, cefpodoxime, and cefotaxim, 96.85% to gentamicin, 89.65% to ceftriaxone, 65.51% to aztreonam, and 40% to piperacillin. Double disk synergy test showed that 21 (28% of non

Process-driven bacterial community dynamics are key to cured meat colour formation by coagulase-negative staphylococci via nitrate reductase or nitric oxide synthase activities.

Science.gov (United States)

Sánchez Mainar, María; Leroy, Frédéric

2015-11-06

The cured colour of European raw fermented meats is usually achieved by nitrate-into-nitrite reduction by coagulase-negative staphylococci (CNS), subsequently generating nitric oxide to form the relatively stable nitrosomyoglobin pigment. The present study aimed at comparing this classical curing procedure, based on nitrate reductase activity, with a potential alternative colour formation mechanism, based on nitric oxide synthase (NOS) activity, under different acidification profiles. To this end, meat models with and without added nitrate were fermented with cultures of an acidifying strain (Lactobacillus sakei CTC 494) and either a nitrate-reducing Staphylococcus carnosus strain or a rare NOS-positive CNS strain (Staphylococcus haemolyticus G110), or by relying on the background microbiota. Satisfactory colour was obtained in the models prepared with added nitrate and S. carnosus. In the presence of nitrate but absence of added CNS, however, cured colour was only obtained when L. sakei CTC 494 was also omitted. This was ascribed to the pH dependency of the emerging CNS background microbiota, selecting for nitrate-reducing Staphylococcus equorum strains at mild acidification conditions but for Staphylococcus saprophyticus strains with poor colour formation capability when the pH decrease was more rapid. This reliance of colour formation on the composition of the background microbiota was further explored by a side experiment, demonstrating the heterogeneity in nitrate reduction of a set of 88 CNS strains from different species. Finally, in all batches prepared with S. haemolyticus G110, colour generation failed as the strain was systematically outcompeted by the background microbiota, even when imposing milder acidification profiles. Thus, when aiming at colour formation through CNS metabolism, technological processing can severely interfere with the composition and functionality of the meat-associated CNS communities, for both nitrate reductase and NOS activities

Monocytes isolated by positive and negative magnetic sorting techniques show different molecular characteristics and immunophenotypic behaviour [version 3; referees: 2 approved

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Jashdeep Bhattacharjee

2018-03-01

Full Text Available Background: Magnetic sorting of cells, based on  microbead conjugated antibodies (Abs, employs positive as well as negative immunomagnetic separation methods, for isolation of a specific cell population. These microbeads are suggested to be nontoxic, biodegradable carriers conjugated to various antibodies. Isolation of cells through positive selection involves the attachment of antibody conjugated microbeads to the cells of interest, followed by their isolation in the presence of a strong magnetic field to obtain higher purity. Negative selection involves attachment of microbead conjugated antibodies to all other cell populations except the cells of interest, which remain untagged. In the present study, we compared the two methods for their effect on functional and immunophenotypic behavior of isolated CD14+ monocytes. Methods: Peripheral blood mononuclear cells (PBMCs were isolated from blood collected from healthy volunteers by density gradient centrifugation. Human blood derived monocytes were isolated through positive selection and negative selection, making use of the appropriate monocyte isolation kit. Monocytes were then stimulated with lipopolysaccharide (LPS and their activation and proliferation capacity were examined. The degradation or dissociation of cell-bound microbeads was also investigated. Results: We observed an impaired LPS sensitivity as well as poor activation and proliferation capacity upon stimulation by LPS in positively sorted CD14+ monocytes as compared to negatively sorted CD14+ monocytes. The attached microbeads did not degrade and remained attached to the cells even after 6 days of culture. Conclusions: Our results suggest that positively sorted CD14+ cells exhibit hampered functionality and may result in inaccurate analysis and observations in downstream applications. However, these cells can be used for immediate analytical procedures.

Antimicrobial resistance of Staphylococcus species isolated from Lebanese dairy-based products.

Science.gov (United States)

Zouhairi, O; Saleh, I; Alwan, N; Toufeili, I; Barbour, E; Harakeh, S

2012-12-04

The study evaluated the antimicrobial resistance of molecularly characterized strains of Staphylococcus aureus and S. saprophyticus isolated from 3 Lebanese dairy-based food products that are sometimes consumed raw: kishk, shanklish and baladi cheese. Suspected Staphylococcus isolates were identified initially using standard biochemical tests, then strains that were confirmed by polymerase chain reaction (29 S. aureus and 17 S. saprophyticus) were evaluated for their susceptibility to different antimicrobials. The highest levels of contamination with staphylococci were in baladi cheese. Resistance rates ranged from 67% to gentamicin to 94% to oxacillin and clindamycin. The results suggest that these locally made dairy-based foods may act as vehicles for the transmission of antimicrobial-resistant Staphylococcus spp.

Worldwide Endemicity of a Multidrug-Resistant Staphylococcus capitis Clone Involved in Neonatal Sepsis.

Science.gov (United States)

Butin, Marine; Martins-Simões, Patricia; Rasigade, Jean-Philippe; Picaud, Jean-Charles; Laurent, Frédéric

2017-03-01

A multidrug-resistant Staphylococcus capitis clone, NRCS-A, has been isolated from neonatal intensive care units in 17 countries throughout the world. S. capitis NRCS-A prevalence is high in some neonatal intensive care units in France. These data highlight the worldwide endemicity and epidemiologic relevance of this multidrug-resistant, coagulase-negative staphylococci clone.

BIOLOGICAL PROPERTIES OF STAPHYLOCOCCUS AND YEAST-LIKE FUNGI OF THE GENUS CANDIDA, ISOLATED FROM ONCOLOGICAL PATIENTS

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Fomina NS

2012-12-01

Full Text Available The results of the spectrum of microorganisms, which are the causative agents of infectious complications of the oral cavity in patients with oncological pathology are presented in the article. The sensitivity of isolated clinical strains of microorganisms to antibiotics, antifungal, antiseptic decasan, miramistin, gorosten, septefril is shown. The results of antiseptic resistance formation to staphylococci and Candida to dekasan, miramistin, septefril, gorosten are described.

BIOLOGICAL PROPERTIES OF STAPHYLOCOCCUS AND YEAST-LIKE FUNGI OF THE GENUS CANDIDA, ISOLATED FROM ONCOLOGICAL PATIENTS

OpenAIRE

Fomina NS; Fomin OO

2012-01-01

The results of the spectrum of microorganisms, which are the causative agents of infectious complications of the oral cavity in patients with oncological pathology are presented in the article. The sensitivity of isolated clinical strains of microorganisms to antibiotics, antifungal, antiseptic decasan, miramistin, gorosten, septefril is shown. The results of antiseptic resistance formation to staphylococci and Candida to dekasan, miramistin, septefril, gorosten are described.

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

, isolates possessed single, bipolar, unsheathed flagella and were urease-negative. They were positive for oxidase and reduced nitrate to nitrite but did not hydrolyse hippurate or indoxyl acetate, grew on charcoal agar and were resistant to cephalothin. 16S rDNA sequences from four strains were determined...

Heterogeneity and phylogenetic relationships of community-associated methicillin-sensitive/resistant Staphylococcus aureus isolates in healthy dogs, cats and their owners.

Science.gov (United States)

Wan, M T; Fu, S Y; Lo, Y P; Huang, T M; Cheng, M M; Chou, C C

2012-01-01

To investigate the distribution of staphylococcal enterotoxin genes (se) and the molecular features of community-associated methicillin-sensitive/resistant Staphylococcus aureus (CA-MSSA/MRSA) isolates in the nostrils of healthy pets and their owners. A total of 114 Staph. aureus isolates were identified from 1563 nasal swab samples, and CA-MRSA accounted for 20·2% (n = 23) of the total identified isolates. CA-MRSA isolates (91·3%, 21/23) harboured higher percentage of se than did CA-MSSA isolates (58·2%, 53/91) (P human bond caused by CA-staphylococci in the commonwealth and the need to take cautions worldwide. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

Prevalence of Staphylococcus aureus Isolated From Various Foods of Animal Origin in Kırşehir, Turkey and Their Enterotoxigenicity

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M. Dilek Avşaroğlu

2016-12-01

Full Text Available The aim of this study was to detect Staphylococcus aureus contamination to different types of animal origin foods collected in the Kırşehir province of Turkey and to examine their enterotoxin production ability. Out of 120 food samples 38 suspected colonies were obtained and 23 of them were identified as S. aureus by biochemical and molecular analyses. Other species detected were S. chromogenes, S. cohnii ssp. cohnii, S. hominis, S. lentus, S. warneri, and S. xylosus. The isolates were also analysed with regard to carry mecA gene. None of them was found to have mecA gene indicating susceptibility to methicillin. To determine the enterotoxigenic ability of the isolates phenotypically, reversed-passive-latex-agglutination test against SEA-SED was used. Six out of 23 S. aureus isolates were determined to produce SEA, SEC and SED. Three of them had only one enterotoxin production, whereas others had SEA and SED production together. The results of phenotypic analyses were confirmed by PCR based examination. None of the coagulase-negative staphylococci were found to be enterotoxigenic by both phenotypical and PCR-based analyses. In conclusion, enterotoxigenic S. aureus is a risk in foods of animal origin in Kırşehir and its counties.

Microbiological and molecular identification of bacterial species isolated from nasal and oropharyngeal mucosa of fuel workers in Riyadh,

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Suaad S. AlWakeel

2017-09-01

Full Text Available This study aimed to determine the bacterial species colonizing the nasal and oropharyngeal mucosa of fuel workers in Central Riyadh, Saudi Arabia on a microbiological and molecular level. Throat and nasal swab samples were obtained from 29 fuel station attendants in the period of time extending from March to May 2014 in Riyadh, Saudi Arabia. Microbiological identification techniques were utilized to identify the bacterial species isolated. Antibiotic sensitivity was assessed for each of the bacterial isolates. Molecular identification techniques based on PCR analysis of specific genomic sequences was conducted and was the basis on which phylogeny representation was done for 10 randomly selected samples of the isolates. Blood was drawn and a complete blood count was conducted to note the hematological indices for each of the study participants. Nineteen bacterial species were isolated from both the nasal cavity and the oropharynx including Streptococcus thoraltensis, alpha-hemolytic streptococci, Staphylococcus hominis, coagulase-negative staphylococci, Leuconostoc mesenteroides, Erysipelothrix rhusiopathiae and several others. We found 100% sensitivity of the isolates to ciprofloxacin, cefuroxime and gentamicin. Whereas cefotaxime and azithromycin posted sensitivities of 85.7% and 91.4%, respectively. Low sensitivities (<60% sensitivity to the antibiotics ampicillin, erythromycin, clarithromycin and norfloxacin were observed. Ninety-seven percent similarity to the microbial bank species was noted when the isolates were compared to it. Most hematological indices recorded were within the normal range. In conclusion, exposure to toxic fumes and compounds within fuel products may be a contributing factor to bacterial colonization of the respiratory tract in fuel workers.

Bovine-associated CNS species resist phagocytosis differently

Science.gov (United States)

2013-01-01

Background Coagulase-negative staphylococci (CNS) cause usually subclinical or mild clinical bovine mastitis, which often remains persistent. Symptoms are usually mild, mostly only comprising slight changes in the appearance of milk and possibly slight swelling. However, clinical mastitis with severe signs has also been reported. The reasons for the differences in clinical expression are largely unknown. Macrophages play an important role in the innate immunity of the udder. This study examined phagocytosis and killing by mouse macrophage cells of three CNS species: Staphylococcus chromogenes (15 isolates), Staphylococcus agnetis (6 isolates) and Staphylococcus simulans (15 isolates). Staphylococcus aureus (7 isolates) was also included as a control. Results All the studied CNS species were phagocytosed by macrophages, but S. simulans resisted phagocytosis more effectively than the other CNS species. Only S. chromogenes was substantially killed by macrophages. Significant variations between isolates were seen in both phagocytosis and killing by macrophages and were more common in the killing assays. Significant differences between single CNS species and S. aureus were observed in both assays. Conclusion This study demonstrated that differences in the phagocytosis and killing of mastitis-causing staphylococci by macrophages exist at both the species and isolate level. PMID:24207012

Distribution of the multidrug resistance gene cfr in Staphylococcus species isolates from swine farms in China.

Science.gov (United States)

Wang, Yang; Zhang, Wanjiang; Wang, Juan; Wu, Congming; Shen, Zhangqi; Fu, Xiao; Yan, Yang; Zhang, Qijing; Schwarz, Stefan; Shen, Jianzhong

2012-03-01

A total of 149 porcine Staphylococcus isolates with florfenicol MICs of ≥ 16 μg/ml were screened for the presence of the multiresistance gene cfr, its location on plasmids, and its genetic environment. In total, 125 isolates carried either cfr (16 isolates), fexA (92 isolates), or both genes (17 isolates). The 33 cfr-carrying staphylococci, which included isolates of the species Staphylococcus cohnii, S. arlettae, and S. saprophyticus in which the cfr gene has not been described before, exhibited a wide variety of SmaI pulsed-field gel electrophoresis patterns. In 18 cases, the cfr gene was located on plasmids. Four different types of cfr-carrying plasmids--pSS-01 (n = 2; 40 kb), pSS-02 (n = 3; 35.4 kb), pSS-03 (n = 10; 7.1 kb), and pBS-01 (n = 3; 16.4 kb)--were differentiated on the basis of their sizes, restriction patterns, and additional resistance genes. Sequence analysis revealed that in plasmid pSS-01, the cfr gene was flanked in the upstream part by a complete aacA-aphD-carrying Tn4001-like transposon and in the downstream part by a complete fexA-carrying transposon Tn558. In plasmid pSS-02, an insertion sequence IS21-558 and the cfr gene were integrated into transposon Tn558 and thereby truncated the tnpA and tnpB genes. The smallest cfr-carrying plasmid pSS-03 carried the macrolide-lincosamide-streptogramin B resistance gene erm(C). Plasmid pBS-01, previously described in Bacillus spp., harbored a Tn917-like transposon, including the macrolide-lincosamide-streptogramin B resistance gene erm(B) in the cfr downstream region. Plasmids, which in part carry additional resistance genes, seem to play an important role in the dissemination of the gene cfr among porcine staphylococci.

eap Gene as novel target for specific identification of Staphylococcus aureus.

Science.gov (United States)

Hussain, Muzaffar; von Eiff, Christof; Sinha, Bhanu; Joost, Insa; Herrmann, Mathias; Peters, Georg; Becker, Karsten

2008-02-01

The cell surface-associated extracellular adherence protein (Eap) mediates adherence of Staphylococcus aureus to host extracellular matrix components and inhibits inflammation, wound healing, and angiogenesis. A well-characterized collection of S. aureus and non-S. aureus staphylococcal isolates (n = 813) was tested for the presence of the Eap-encoding gene (eap) by PCR to investigate the use of the eap gene as a specific diagnostic tool for identification of S. aureus. Whereas all 597 S. aureus isolates were eap positive, this gene was not detectable in 216 non-S. aureus staphylococcal isolates comprising 47 different species and subspecies of coagulase-negative staphylococci and non-S. aureus coagulase-positive or coagulase-variable staphylococci. Furthermore, non-S. aureus isolates did not express Eap homologs, as verified on the transcriptional and protein levels. Based on these data, the sensitivity and specificity of the newly developed PCR targeting the eap gene were both 100%. Thus, the unique occurrence of Eap in S. aureus offers a promising tool particularly suitable for molecular diagnostics of this pathogen.

Browse Title Index

African Journals Online (AJOL)

Items 301 - 350 of 458 ... F.C. Thomas, O.S. Aduloju, B Okediran, E.S. Ajibola, E.N. Maduagwu. Vol 37, No 3 (2016), Prevalence and antimicrobial resistance pattern of coagulase negative Staphylococci isolated from pigs and in-contact humans in Jos Metropolis, Nigeria, Abstract PDF. A.H. Momoh, J. K. P. Kwaga, M. Bello, ...

Aerobic bacteria from mucous membranes, ear canals, and skin wounds of feral cats in Grenada, and the antimicrobial drug susceptibility of major isolates.

Science.gov (United States)

Hariharan, Harry; Matthew, Vanessa; Fountain, Jacqueline; Snell, Alicia; Doherty, Devin; King, Brittany; Shemer, Eran; Oliveira, Simone; Sharma, Ravindra N

2011-03-01

In a 2-year period 54 feral cats were captured in Grenada, West Indies, and a total of 383 samples consisting of swabs from rectum, vagina, ears, eyes, mouth, nose and wounds/abscesses, were cultured for aerobic bacteria and campylobacters. A total of 251 bacterial isolates were obtained, of which 205 were identified to species level and 46 to genus level. A commercial bacterial identification system (API/Biomerieux), was used for this purpose. The most common species was Escherichia coli (N=60), followed by Staphylococcus felis/simulans (40), S. hominis (16), S. haemolyticus (12), Streptococcus canis (9), Proteus mirabilis (8), Pasteurella multocida (7), Streptococcus mitis (7), Staphylococcus xylosus (7), S. capitis (6), S. chromogenes (4), S. sciuri (3), S. auricularis (2), S. lentus (2), S. hyicus (2), Streptococcus suis (2) and Pseudomonas argentinensis (2). Sixteen other isolates were identified to species level. A molecular method using 16S rRNA sequencing was used to confirm/identify 22 isolates. Salmonella or campylobacters were not isolated from rectal swabs. E. coli and S. felis/simulans together constituted 50% of isolates from vagina. S. felis/simulans was the most common species from culture positive ear and eye samples. P. multocida was isolated from 15% of mouth samples. Coagulase-negative staphylococci were the most common isolates from nose and wound swabs. Staphylococcus aureus, or S. intemedius/S. pseudintermedius were not isolated from any sample. Antimicrobial drug resistance was minimal, most isolates being susceptible to all drugs tested against, including tetracycline. Copyright © 2010 Elsevier Ltd. All rights reserved.

Microbiota and anthropic interference on antimicrobial resistance profile of bacteria isolated from Brazilian maned-wolf (Chrysocyon brachyurus

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Olney Vieira-da-Motta

2013-12-01

Full Text Available Both the study of Brazilian wild mammal fauna and the conditions that foster the preservation of endangered species, such as Brazilian Maned-wolf (Chrysocyon brachyurus, in wild life are of extreme importance. In order to study the resistance profile of microbiota bacterial colonizing Brazilian Maned-wolf, this work investigated samples from eight male captive and free roaming animals originating from different Brazilian geographical regions. Samples for microbiological purposes were collected with swabs and kept in appropriate transport medium. Using routine microbiological techniques, the isolated bacteria were tested toward antimicrobial drugs by the agar disk diffusion method. Results showed that all samples from wild animals were sensitive toward all drugs tested. Conversely, the resistance profile of bacteria isolated from captive animals varied among strains and animal body site location. Escherichia coli samples from prepuce, anus and ear showed multi-resistance toward at least four drugs, especially against erythromycin and tetracycline, followed by Proteus mirabilis and P. vulgaris strains isolated from anus and ear. Among Gram-positive bacteria, strains of coagulase-negative staphylococci showed multi-resistance mainly toward erythromycin and amoxicillin. The work discusses these findings and suggests that profile of multi-resistance bacteria from captive subjects may be attributed to direct contact with human or through lifestyle factors such as feeding, predation or contact of animals with urban animals such as birds, rodents, and insects from surrounding environments.

Microbiota and anthropic interference on antimicrobial resistance profile of bacteria isolated from Brazilian Maned-wolf (Chrysocyon brachyurus).

Science.gov (United States)

Vieira-da-Motta, Olney; Eckhardt-de-Pontes, Luiz Antonio; Petrucci, Melissa Paes; dos Santos, Israel Pereira; da Cunha, Isabel Candia Nunes; Morato, Ronaldo Gonçalves

2013-12-01

Both the study of Brazilian wild mammal fauna and the conditions that foster the preservation of endangered species, such as Brazilian Maned-wolf (Chrysocyon brachyurus), in wild life are of extreme importance. In order to study the resistance profile of microbiota bacterial colonizing Brazilian Maned-wolf, this work investigated samples from eight male captive and free roaming animals originating from different Brazilian geographical regions. Samples for microbiological purposes were collected with swabs and kept in appropriate transport medium. Using routine microbiological techniques, the isolated bacteria were tested toward antimicrobial drugs by the agar disk diffusion method. Results showed that all samples from wild animals were sensitive toward all drugs tested. Conversely, the resistance profile of bacteria isolated from captive animals varied among strains and animal body site location. Escherichia coli samples from prepuce, anus and ear showed multi-resistance toward at least four drugs, especially against erythromycin and tetracycline, followed by Proteus mirabilis and P. vulgaris strains isolated from anus and ear. Among Gram-positive bacteria, strains of coagulase-negative staphylococci showed multi-resistance mainly toward erythromycin and amoxicillin. The work discusses these findings and suggests that profile of multi-resistance bacteria from captive subjects may be attributed to direct contact with human or through lifestyle factors such as feeding, predation or contact of animals with urban animals such as birds, rodents, and insects from surrounding environments.

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.

Staphylococcus aureus penetrate the interkeratinocyte spaces created by skin-infiltrating neutrophils in a mouse model of impetigo.

Science.gov (United States)

Imanishi, Ichiro; Hattori, Shinpei; Hisatsune, Junzo; Ide, Kaori; Sugai, Motoyuki; Nishifuji, Koji

2017-02-01

Impetigo is a bacterial skin disease characterized by intraepidermal neutrophilic pustules. Previous studies have demonstrated that exfoliative toxin producing staphylococci are isolated in the cutaneous lesions of human and canine impetigo. However, the mechanisms of intraepidermal splitting in impetigo remain poorly understood. To determine how staphylococci penetrate the living epidermis and create intraepidermal pustules in vivo using a mouse model of impetigo. Three Staphylococcus aureus strains harbouring the etb gene and three et gene negative strains were epicutaneously inoculated onto tape-stripped mouse skin. The skin samples were subjected to time course histopathological and immunofluorescence analyses to detect intraepidermal neutrophils and infiltrating staphylococci. To determine the role of neutrophils on intraepidermal bacterial invasion, cyclophosphamide (CPA) was injected intraperitoneally into the mice to cause leucopenia before the inoculation of etb gene positive strains. In mice inoculated with etb gene positive S. aureus, intraepidermal pustules resembling impetigo were detected as early as 4 h post-inoculation (hpi). Neutrophils in the epidermis were detected from 4 hpi, whereas intraepidermal staphylococci was detected from 6 hpi. The dimensions of the intraepidermal clefts created in mice inoculated with etb gene positive strains at 6 hpi were significantly larger than those in mice inoculated with et gene negative strains. In CPA treated mice, staphylococci or neutrophils were not detected in the deep epidermis until 6 hpi. Our findings indicate that intraepidermal neutrophils play an important role in S. aureus invasion into the living epidermis in a mouse model of impetigo. © 2016 ESVD and ACVD.

Susceptibility of different bacterial species isolated from food animals to copper sulphate, zinc chloride and antimicrobial substances used for disinfection

DEFF Research Database (Denmark)

Aarestrup, Frank Møller; Hasman, Henrik

2004-01-01

that Danish bacterial isolates from livestock so far have not or have only to a limited degree developed resistance to antimicrobial compounds commonly used for disinfection. Acquired copper resistance was only found in enterococci. There were large differences in the intrinsic susceptibility of the different...... of susceptibilities to the different antimicrobial agents. Large variations were observed in the susceptibility of the different bacterial species to the different compounds. Staphylococci were in general very susceptible to all antimicrobial compounds tested. The Salmonella isolates were in general less susceptible...

Antimicrobial susceptibility of coagulase-negative Staphylococcus species isolated from bovine milk.

Science.gov (United States)

Sawant, A A; Gillespie, B E; Oliver, S P

2009-02-16

Coagulase-negative Staphylococcus (CNS) isolates (n=168) obtained from milk from heifers and dairy cows were screened for minimum inhibitory concentration (MIC) to antimicrobials used commonly for mastitis therapy. Of the 10 CNS species included in the study, the predominant species were Staphylococcus chromogenes (n=61), Staphylococcus epidermidis (n=37), Staphylococcus hyicus (n=37), and Staphylococcus simulans (n=16). The majority of CNS was susceptible to ampicillin, oxacillin, cephalothin, and ceftiofur. Erythromycin and pirlimycin were also very effective in vitro inhibitors of CNS. The only exception was observed with S. epidermidis. Of 37 S. epidermidis evaluated, 13 (35%) exhibited efflux-based resistance to erythromycin (> or =16 microg/ml) encoded by msrA and one isolate carried ermC encoding ribosomal methylase-based resistance to both erythromycin (> or =64 microg/ml) and pirlimycin (> or =64 microg/ml). A total of 17 S. epidermidis, 11 S. chromogenes, and one S. hyicus exhibited phenotypic resistance to ampicillin (> or =0.5 microg/ml). Constitutive beta-lactamase production was observed in all ampicillin resistant isolates except 4 S. epidermidis that exhibited inducible beta-lactamase production. Induced beta-lactamase production was also observed in 13 S. epidermidis that were phenotypically susceptible to the entire MIC panel. All isolates that produced beta-lactamase either constitutively or by induction carried blaZ. S. epidermidis (n=12, 32%) that were resistant to methicillin (oxacillin > or =0.5 microg/ml) carried low affinity penicillin-binding protein encoded by mecA. Most multi-drug resistant (MDR) S. epidermidis (> or =2 resistance genes) were resistant to ampicillin, erythromycin and methicillin. All except one MDR S. epidermidis had icaAB, which encodes for polysaccharide intercellular adhesion. Based on pulsed field gel electrophoresis, MDR S. epidermidis were closely related genotypically, and were isolated from different cows on the

Multicenter Evaluation of the Bruker MALDI Biotyper CA System for the Identification of Clinical Aerobic Gram-Negative Bacterial Isolates.

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Matthew L Faron

Full Text Available The prompt and accurate identification of bacterial pathogens is fundamental to patient health and outcome. Recent advances in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS have revolutionized bacterial identification in the clinical laboratory, but uniform incorporation of this technology in the U.S. market has been delayed by a lack of FDA-cleared systems. In this study, we conducted a multicenter evaluation of the MALDI Biotyper CA (MBT-CA System (Bruker Daltonics Inc, Billerica, MA for the identification of aerobic gram-negative bacteria as part of a 510(k submission to the FDA. A total of 2,263 aerobic gram negative bacterial isolates were tested representing 23 genera and 61 species. Isolates were collected from various clinical sources and results obtained from the MBT-CA System were compared to DNA sequencing and/or biochemical testing. Isolates that failed to report as a "high confidence species ID" [log(score ≥2.00] were re-tested using an extraction method. The MBT-CA System identified 96.8% and 3.1% of isolates with either a "high confidence" or a "low confidence" [log(score value between 1.70 and <2.00] species ID, respectively. Two isolates did not produce acceptable confidence scores after extraction. The MBT-CA System correctly identified 99.8% (2,258/2,263 to genus and 98.2% (2,222/2,263 to species level. These data demonstrate that the MBT-CA System provides accurate results for the identification of aerobic gram-negative bacteria.

Multicenter Evaluation of the Bruker MALDI Biotyper CA System for the Identification of Clinical Aerobic Gram-Negative Bacterial Isolates.

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Faron, Matthew L; Buchan, Blake W; Hyke, Josh; Madisen, Neil; Lillie, Jennifer L; Granato, Paul A; Wilson, Deborah A; Procop, Gary W; Novak-Weekley, Susan; Marlowe, Elizabeth; Cumpio, Joven; Griego-Fullbright, Christen; Kindig, Sandra; Timm, Karen; Young, Stephen; Ledeboer, Nathan A

2015-01-01

The prompt and accurate identification of bacterial pathogens is fundamental to patient health and outcome. Recent advances in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) have revolutionized bacterial identification in the clinical laboratory, but uniform incorporation of this technology in the U.S. market has been delayed by a lack of FDA-cleared systems. In this study, we conducted a multicenter evaluation of the MALDI Biotyper CA (MBT-CA) System (Bruker Daltonics Inc, Billerica, MA) for the identification of aerobic gram-negative bacteria as part of a 510(k) submission to the FDA. A total of 2,263 aerobic gram negative bacterial isolates were tested representing 23 genera and 61 species. Isolates were collected from various clinical sources and results obtained from the MBT-CA System were compared to DNA sequencing and/or biochemical testing. Isolates that failed to report as a "high confidence species ID" [log(score) ≥2.00] were re-tested using an extraction method. The MBT-CA System identified 96.8% and 3.1% of isolates with either a "high confidence" or a "low confidence" [log(score) value between 1.70 and <2.00] species ID, respectively. Two isolates did not produce acceptable confidence scores after extraction. The MBT-CA System correctly identified 99.8% (2,258/2,263) to genus and 98.2% (2,222/2,263) to species level. These data demonstrate that the MBT-CA System provides accurate results for the identification of aerobic gram-negative bacteria.

Phage type and sensitivity to antibiotics of Staphylococcus aureus film-forming strains isolated from airway mucosa

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O. S. Voronkova

2014-10-01

Full Text Available Today film-forming strains of bacteria play very important role in clinical pathology. Staphylococci are ones of most dangerous of them. This bacteria can determine different pathological processes, for example, complication of airway mucosa. The ability to form a biofilm is one of the main properties of nosocomial strains. These strains should be monitored and their carriers are to be properly treated. To determine the origin of staphylococci strains we used bacteriophages from the International kit. The aim of research was to determine the phage type of staphylococci film-forming strains, that were isolated from naso-pharingial mucosa. Phage typing has been carried out for 16 film-forming strains of S. aureus. To solve this problem, we used the International phage kit by Fisher’s method. As a result, sensitivity to phages from the International kit showed 53.8% of studied strains of S. aureus. 64.3% of sensitivity strains were lysed by one of the phage, 21.4% – were by two of the phages, 14.3% – by three of the phages. Isolates were sensitive to phages: 81 – 42.9%, 75 – 35.7%, 28.6% were sensitive to phages 47 and 53. All cases of detection of sensitivity to phage 47 coincided with the ability to form biofilm. Among non-film-forming strains there was no sensitive strains for this phage. Film-forming strains resist to erythromycin (62.5%, ciprofloxacin (43.8%, gentamicin (56.3%, tetracycline (87.5%, amoxicillin (93.8%, and cefuroxime (37.5%. All cases of sensitivity to phage 47 coincided with resistance to erythromycin, amoxicillin and tetracycline. For two of these strains, we also defined resistance to gentamicin and for one of them – to ciprofloxacin. Results of research allowed to relate the bacterial cultures for determining the type. This may have implications for studying of film-forming ability, because surface structures of bacterial cell take place in this process. Belonging of an isolate to specific phage type may

Antibiotic susceptibility of methicillin-resistant staphylococci (MRS) of food origin: A comparison of agar disc diffusion method and a commercially available miniaturized test.

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Buzón-Durán, Laura; Capita, Rosa; Alonso-Calleja, Carlos

2018-06-01

Methicillin-resistant staphylococci (MRS) are a major concern to public and animal health. Thirty MRS (Staphylococcus aureus, S. cohnii, S. epidermidis, S. haemolyticus, S. hominis, S. lentus, S. lugdunensis, S. sciuri, and S. xylosus) isolates from meat and poultry preparations were tested for antimicrobial susceptibility to 11 antimicrobials (belonging to seven different categories) of clinical significance using both the standard agar disc diffusion method and a commercially available miniaturized system (Sensi Test Gram-positive). It is worth stressing that 16 isolates (53.33%) exhibited an extensively drug-resistant phenotype (XDR). The average number of resistances per strain was 4.67. These results suggest that retail meat and poultry preparations are a likely vehicle for the transmission of multi-drug resistant MRS. Resistance to erythromycin was the commonest finding (76.67% of strains), followed by tobramycin, ceftazidime (66.67%), ciprofloxacin (56.67%) and fosfomycin (53.33%). An agreement (kappa coefficient) of 0.64 was found between the two testing methods. Using the agar disc diffusion as the reference method, the sensitivity, specificity and accuracy of the miniaturized test were 98.44%, 69.44% and 83.33%, respectively. Most discrepancies between the two methods were due to isolates that were susceptible according to the disc diffusion method but resistant according to the miniaturized test (false positives). Copyright © 2017. Published by Elsevier Ltd.

Direct Identification and Antimicrobial Susceptibility Testing of Bacteria From Positive Blood Culture Bottles by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry and the Vitek 2 System.

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Jo, Sung Jin; Park, Kang Gyun; Han, Kyungja; Park, Dong Jin; Park, Yeon-Joon

2016-03-01

We evaluated the reliability and accuracy of the combined use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) bacterial identification and Vitek 2 antimicrobial susceptibility testing (AST) for bacteria from positive blood culture bottles. Direct identification and AST were performed in parallel to the standard methods in monomicrobial positive blood culture bottles. In total, 254 isolates grown on aerobic and/or anaerobic bottles were identified with MALDI-TOF Vitek MS (bioMérieux, France), and 1,978 microorganism/antimicrobial agent combinations were assessed. For isolates from anaerobic bottles, an aliquot of the culture broth was centrifuged, washed, and filtered through a nylon mesh. For isolates from aerobic/pediatric bottles, a lysis step using 9.26% ammonium chloride solution and 2% saponin solution was included. The overall correct identification rate was 81.8% (208/254) and that for gram-positive/gram-negative isolates was 73.9%/92.6%, respectively, and it was 81.8%, 87.6%, and 57.9% for isolates from aerobic, anaerobic, and pediatric bottles, respectively. Identification was not possible in 45 cases, and most of these isolates were streptococci (N=14) and coagulase-negative staphylococci (N=11). Misidentification occurred only in one case. Compared with standard methods, direct AST showed 97.9% (1,936/1,978) agreement with very major error of 0.25%, major error of 0.05%, and minor error of 1.8%. This simple and cost-effective sample preparation method gives reliable results for the direct identification and AST of bacteria. For the identification of streptococci and coagulase-negative staphylococci, the method should be further improved.

Antibiotic exposure in a low-income country: screening urine samples for presence of antibiotics and antibiotic resistance in coagulase negative staphylococcal contaminants.

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Anne Mette Lerbech

Full Text Available 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 (CoNS are suggested to evolve due to positive selective pressure following antibiotic treatment. This study investigated the presence of the nine most commonly used antimicrobial agents in human urine from outpatients in two hospitals in Ghana in relation to CoNS resistance. Urine and CoNS were sampled (n = 246 and n = 96 respectively from patients in two hospitals in Ghana. CoNS were identified using Gram staining, coagulase test, and MALDI-TOF/MS, and the antimicrobial susceptibility to 12 commonly used antimicrobials was determined by disk diffusion. Moreover an analytical method was developed for the determination of the nine most commonly used antimicrobial agents in Ghana by using solid-phase extraction in combination with HPLC-MS/MS using electron spray ionization. The highest frequency of resistance to CoNS was observed for penicillin V (98%, trimethoprim (67%, and tetracycline (63%. S. haemolyticus 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 agents were detected in 64% of the analysed urine samples (n = 121 where the most frequently detected antimicrobials were ciprofloxacin (30%, trimethoprim (27%, and metronidazole (17%. The major findings of this study was that the prevalence of detected antimicrobials in urine was more frequent than the use reported by the patients and the prevalence of resistant S. haemolyticus was more frequent than other resistant CoNS species when antimicrobial

Microbiological analysis and sensitivity profile of Staphylococcus spp. in subclinical mastitis of dairy goats

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S.R.S. Salaberry

2016-04-01

Full Text Available Subclinical mastitis in goats causes economic losses and risks to public health. Given the need for research that shows the most isolated staphylococci species and sensibility tests comparing the resistance between coagulase-negative (CNS and positive Staphylococcus (CPS goats with subclinical mastitis, the aim of this study was to identify the microorganisms isolated from milk samples of goats with subclinical mastitis, as well as define the staphylococci species and determine the sensitivity profile of Staphylococcus spp. to antimicrobials. To collect samples, tests were performed for mug of black background and California mastitis test (CMT, collecting milk from CMT positive animals. A total of 226 samples from seven herds of dairy goats was collected and forwarded to the laboratory, where they were seeded for the isolation of the microorganism and implementing the antibiotic sensibility test. Of these, 122 samples had bacterial growth and the most isolated staphylococci species were: S. epidermidis (24.55%, S. lugdunensis (15.40% and S. intermedius (13.64%. Samples showed increased resistance to antimicrobials: penicillin (81.8%, oxacillin (60.0% and ampicillin (55.5%. Greater sensitivity to: enrofloxacin (99.1%, erythromycin (98.2%, gentamicin (98.2% and vancomycin (98.2% were observed. The S. epidermidis showed higher antimicrobial resistance to amoxicillin and penicillin than S. lugdunensis and S. intermedius. Similar resistance in vitro between CNS and CPS was observed to most antimicrobials. It is important to control the overuse of antibiotics to prevent the emergence of resistant strains.

Biofilm Formation and Antimicrobial Susceptibility of Staphylococcus epidermidis Strains from a Hospital Environment

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Robert D. Wojtyczka

2014-04-01

Full Text Available The hospital environment microflora comprise a wide variety of microorganisms which are more or less pathogenic and where staphylococci are one of the most common types. The aim of the presented study was to evaluate the prevalence of the biofilm forming coagulase-negative staphylococci (CoNS in a hospital environment as a risk factor for nosocomial infections. Among 122 isolated and tested strains of CoNS the most frequent were: S. epidermidis—32 strains, S. haemolyticus—31 strains, S. capitis subsp. capitis— 21 strains, S. hominis—11 strains, S. cohnii subsp. cohnii—nine strains. In case of CoNS, the main molecule responsible for intercellular adhesion is a polysaccharide intercellular adhesin (PIA, encoded on the ica gene operon. The analysis revealed the presence of the icaADBC operon genes in 46.88% of S. epidermidis isolates. IcaA and icaD were present in 34.38% and 28.13% of strains respectively while IcaC gene was present in 37.50% of strains. IcaB gene was found in 21.88% of S. epidermidis strains. In 15 (63% strains all icaADBC operon genes were observed. The assessment of antibacterial drugs susceptibility demonstrated that analyzed CoNS strains were highly resistant to macrolides and lincosamides and more sensitive to rifampicin and linezolid. Our data indicates that the hospital environment can be colonized by biofilm forming coagulase-negative staphylococci and transmission of these strains can cause an increased risk of serious nosocomial infections.

Biofilm formation and antimicrobial susceptibility of Staphylococcus epidermidis strains from a hospital environment.

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Wojtyczka, Robert D; Orlewska, Kamila; Kępa, Małgorzata; Idzik, Danuta; Dziedzic, Arkadiusz; Mularz, Tomasz; Krawczyk, Michał; Miklasińska, Maria; Wąsik, Tomasz J

2014-04-25

The hospital environment microflora comprise a wide variety of microorganisms which are more or less pathogenic and where staphylococci are one of the most common types. The aim of the presented study was to evaluate the prevalence of the biofilm forming coagulase-negative staphylococci (CoNS) in a hospital environment as a risk factor for nosocomial infections. Among 122 isolated and tested strains of CoNS the most frequent were: S. epidermidis-32 strains, S. haemolyticus-31 strains, S. capitis subsp. capitis- 21 strains, S. hominis-11 strains, S. cohnii subsp. cohnii-nine strains. In case of CoNS, the main molecule responsible for intercellular adhesion is a polysaccharide intercellular adhesin (PIA), encoded on the ica gene operon. The analysis revealed the presence of the icaADBC operon genes in 46.88% of S. epidermidis isolates. IcaA and icaD were present in 34.38% and 28.13% of strains respectively while IcaC gene was present in 37.50% of strains. IcaB gene was found in 21.88% of S. epidermidis strains. In 15 (63%) strains all icaADBC operon genes were observed. The assessment of antibacterial drugs susceptibility demonstrated that analyzed CoNS strains were highly resistant to macrolides and lincosamides and more sensitive to rifampicin and linezolid. Our data indicates that the hospital environment can be colonized by biofilm forming coagulase-negative staphylococci and transmission of these strains can cause an increased risk of serious nosocomial infections.

Impaired glucocorticoid-mediated HPA axis negative feedback induced by juvenile social isolation in male rats.

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Boero, Giorgia; Pisu, Maria Giuseppina; Biggio, Francesca; Muredda, Laura; Carta, Gianfranca; Banni, Sebastiano; Paci, Elena; Follesa, Paolo; Concas, Alessandra; Porcu, Patrizia; Serra, Mariangela

2018-05-01

We previously demonstrated that socially isolated rats at weaning showed a significant decrease in corticosterone and adrenocorticotropic hormone (ACTH) levels, associated with an enhanced response to acute stressful stimuli. Here we shown that social isolation decreased levels of total corticosterone and of its carrier corticosteroid-binding globulin, but did not influence the availability of the free active fraction of corticosterone, both under basal conditions and after acute stress exposure. Under basal conditions, social isolation increased the abundance of glucocorticoid receptors, while it decreased that of mineralocorticoid receptors. After acute stress exposure, socially isolated rats showed long-lasting corticosterone, ACTH and corticotrophin releasing hormone responses. Moreover, while in the hippocampus and hypothalamus of group-housed rats glucocorticoid receptors expression increased with time and reached a peak when corticosterone levels returned to basal values, in socially isolated rats expression of glucocorticoid receptors did not change. Finally, social isolation also affected the hypothalamic endocannabinoid system: compared to group-housed rats, basal levels of anandamide and cannabinoid receptor type 1 were increased, while basal levels of 2-arachidonoylglycerol were decreased in socially isolated rats and did not change after acute stress exposure. The present results show that social isolation in male rats alters basal HPA axis activity and impairs glucocorticoid-mediated negative feedback after acute stress. Given that social isolation is considered an animal model of several neuropsychiatric disorders, such as generalized anxiety disorder, depression, post-traumatic stress disorder and schizophrenia, these data could contribute to better understand the alterations in HPA axis activity observed in these disorders. Copyright © 2018 Elsevier Ltd. All rights reserved.

Prevalence and antibiotic susceptibility of coagulase-negative Staphylococcus species from bovine subclinical mastitis in dairy herds in the central region of Argentina.

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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. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

Interaction of Staphylococci with Human B cells.

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Tyler K Nygaard

Full Text Available Staphylococcus aureus is a leading cause of human infections worldwide. The pathogen produces numerous molecules that can interfere with recognition and binding by host innate immune cells, an initial step required for the ingestion and subsequent destruction of microbes by phagocytes. To better understand the interaction of this pathogen with human immune cells, we compared the association of S. aureus and S. epidermidis with leukocytes in human blood. We found that a significantly greater proportion of B cells associated with S. epidermidis relative to S. aureus. Complement components and complement receptors were important for the binding of B cells with S. epidermidis. Experiments using staphylococci inactivated by ultraviolet radiation and S. aureus isogenic deletion mutants indicated that S. aureus secretes molecules regulated by the SaeR/S two-component system that interfere with the ability of human B cells to bind this bacterium. We hypothesize that the relative inability of B cells to bind S. aureus contributes to the microbe's success as a human pathogen.

Contaminant bacteria in traditional-packed honey

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Hening Tjaturina Pramesti

2007-03-01

Full Text Available Honey may be contaminated by microorganisms during its harvesting, processing, and packaging. Honey selected for clinical purposes must safe, sterile, and contain antimicrobial activity, so it must be evaluated using laboratory testing. The aim of this descriptive laboratory study was to isolate and identify the bacterial contaminant in the traditional-packed honey dealing with the use of honey for medical purposes. the colony forming units of honey sample cultured on blood agar were counted using Stuart bacterial colony counter. The suspected bacterial colonies were isolated and identified based on cultural morphology characteristics. The isolates of suspected bacterial colonies were stained according to Gram and Klein method and then were examined by the biochemical reaction. The results showed that there were two contaminant bacteria. Gram-positive cocci which were presumptively identified as coagulase-negative Staphylococci and gram-positive rods which were presumptively identified as Bacillus subtilis. In conclusion, the contaminant bacteria were regarded as low pathogen bacteria. The subtilin enzyme of B subtilis may cause an allergic reaction and coagulase-negative Staphylococci, Staphylococcus epidermidis is also an opportunist pathogen. Inevitably, for medical purposes, traditional-packed honey must be well filtered, water content above 18%, and standardized sterilization without loss of an antibacterial activity or change in properties.

Incidenza della meticillino-resistenza in Staphylococcus aureus e stafilococchi coagulasi-negativi isolati da emocolture

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Alessandra Siddi

2007-12-01

Full Text Available Background: Staphylococci are major cause of nosocomial blood stream infections.This local surveillance study was carry out to monitor frequency of occurrence of Staphylococcus aureus and coagulase-negative staphylococci (CoNS in blood stream infections and the incidence of methicillin-resistant (MET-R strains. Materials and methods: During the period January – December 2006, 9840 blood specimens were analyzed and microrganisms from positive samples were collected. Bacterial identifications were performed according to the standard methods (Murray, 2003.We evaluated, in particular, the antibiotic-resistance phenotype of staphylococci employing disk diffusion test as suggested by the CLSI (2006. The following antimicrobial agents were tested: oxacillin, penicillin, amoxiciclin-clavulanate, cefalotin, cefamandole, imipenem, teicoplanin, linezolid, ciprofloxacin, erythromycin, clindamicin, rifampicin, trimethoprim-sulfamethoxazole, gentamicin, doxiciclin, fosfomycin. Results: The microrganisms isolated were 551: 370 Gram-positives (67%, 131 Gram-negatives (24%, 11 anaerobes (2% and 39 mycetes (7%. In particular, 121 S. epidermidis, 75 S. aureus, 42 S. haemolyticus and other 39 CoNS were analyzed: methicillin-resistance occurred in more than 80% of S.aureus strains collected from Intensive Care Units (ICU and in about 50 % of those isolated from other divisions. In CoNS the incidence of MET-R ranged from 30 to 80 %, the higher values were registered among S. epidermidis and S. haemolyticus. MET-R strains were characterized by high resistance rates even to ciprofloxacin (from 47 to 100%, erythromycin (from 70 to 100%, and in same cases to gentamicin (from 23 to 86% also. Conclusions: Staphylococci are the prevalent cause of blood stream infections.The distinctive feature of MET-R strains is their resistance not only to all b-lactam antibiotics, but also to a wide range of other antimicrobial agents. However, the glycopeptide teicoplanin remains 100

Staphylococcus species and their Methicillin-Resistance in 7424 Blood Cultures for Suspected Bloodstream Infections

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Ariana ALMAŞ

2011-06-01

Full Text Available Objectives: The aim of this study was to evaluate the distribution of Staphylococcus species in bloodstream infections and to assess their susceptibility to methicillin. Material and Methods: Between January 1st 2008 - December 31st 2010, 7424 blood culture sets were submitted to the Laboratory Department of the Hospital for Clinical Infectious Diseases in Cluj-Napoca, Romania. The blood cultures were performed using BacT/Alert until January 2010 and BacT/Alert 3D automated system (bioMérieux after that date. The blood culture bottles were incubated at 37°C in a continuously monitoring system for up to 7 days. The strain identifications were performed by conventional methods, ApiStaph galleries and Vitek 2 Compact system. Susceptibility to methicillin was determined by disk diffusion method with cefoxitin disk and by using Vitek 2 Compact system. Results: From the total number of performed blood cultures, 568 were positive with Staphylococcus species. From 168 bacteriemic episodes 103 were with Staphylococcus aureus. Among 65 coagulase-negative staphylococci isolates, Staphylococcus epidermidis was the most frequently isolated species (34, followed by Staphylococcus hominis (15, Staphylococcus haemolyticus (8, Staphylococcus saprophyticus (3, Staphylococcus cohnii (1, Staphylococcus auricularis (1, and 3 strains that were not identified at species level. Methicillin resistance was encountered in 53.40% of Staphylococcus aureus strains and in 80% of coagulase-negative staphylococci. Conclusions: An important percentage of blood cultures were contaminated with Staphylococcus species. The main species identified in true bacteriemia cases were Staphylococcus aureus and Staphylococcus epidermidis. The percentage of methicillin-resistance, proved to be high not only for coagulase-negative staphylococci but also for Staphylococcus aureus.

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

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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.

Bacterial subclinical mastitis and its effect on milk yield in low-input dairy goat herds.

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Gelasakis, A I; Angelidis, A S; Giannakou, R; Filioussis, G; Kalamaki, M S; Arsenos, G

2016-05-01

The objectives of this study were (1) to record the major pathogens associated with subclinical mastitis (SCM), (2) to calculate their incidence during the milking period, and (3) to estimate the effect of SCM on daily milk yield (DMY) for goats reared under low-input management schemes. Dairy goats (n=590) of Skopelos and indigenous Greek breeds from 4 herds were randomly selected for the study. The study included monthly monitoring, milk yield recording, and bacteriological analyses of milk of individual goats during the course of 2 successive milking periods. Incidence and cumulative incidence were calculated for SCM cases. Moreover, 2 mixed linear regression models were built to assess the effects of (1) SCM and (2) different pathogens isolated from SCM cases, on DMY. The estimated incidence and cumulative incidence of SCM for the first and the second year of the study were 69.5 and 96.4 new cases of SCM/1,000 goat-months, and 24.1 and 31.7%, respectively. A total of 755 milk samples were subjected to microbiological examination, resulting in 661 positive cultures. Coagulase-negative and coagulase-positive staphylococci were isolated from 50.2 and 34.5% of the positive cultures, respectively. The incidence of infections (new infections per 1,000 goat-months) for the first and the second year of the study were 34 and 53 for coagulase-negative staphylococci, 23 and 28 for coagulase-positive staphylococci, 3 and 5 for Streptococcus/Enterococcus spp., and 5.5 and 9.1 for gram-negative bacteria. Goats with SCM had lower DMY when compared with goats without SCM (ca. 47g/d, corresponding to a 5.7% decrease in DMY). In particular, goats with SCM due to coagulase-positive staphylococci infection produced approximately 80g/d less milk (a reduction of ca. 9.7%) compared with uninfected ones, whereas SCM due to gram-negative bacteria resulted in approximately 15% reduction in DMY. Investigating the epidemiology of SCM and its effects on production traits is critical for

Investigate Nasal Colonize Staphylococcus Species Biofilm Produced

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Cemil Demir

2014-03-01

Full Text Available Aim: 127 S.aureus and 65 CoNS strains were isolated from patients noses%u2019. To produce a biofilm ability was investigated using three different methods. Slime-positive and negative staphylococcies%u2019 resistance were evaluated against different antibiotics. Material and Method: Swap samples puted 7% blood agar. Staphylococcus aureus and coagulase-negative staphylococci (CoNS isolates biofilm produced ability were investigated using Congo Red Agar (CRA, microplates (MP and Standard Tube (ST methods. In addition to that, presence of antibiotic resistance of the staphylococcal isolates are determined agar disc diffusion method. Results: The rate of biofilm producing Staphylococcus spp strains was found to be 72.4%, 67.7%, and 62.9%, respectively with CRA, MP, and ST tests. There was no significant relationship among the tests (p>0.05. In addition, antibiotic resistance of Staphylococcus spp. against various antibiotics was also determined by the agar disk diffusion method. Resistance rates of biofilm positive (BP Staphylococcus spp for penicilin G, ampicilin, amocycilin/clavulanic acid, tetracyclin, eritromycin, gentamycin, and enrofloxacin 71.7%, 69.7%, 6.2%, 20.7%, 21.4%, 1.4%, and 0.7%, respectively. Resistance rates of biofilm negative (BN spp for 42.6%, 23.4%, 4.3%, 14.9%, 19.1%, 0.0%, 0.0% respectively. All Staphylococcus isolates were found to be susceptible to vancomycin and teicaplonin. Although BP strains antibiotic resistance rates were observed higher than BN strains. But resistance rates were not found statistically significant (p>0.05. Discussion: CRA is the reliablity and specifity method to determine Staphylococcus spp. biofilm produce ability.

Efficacy of a marine bacterial nuclease against biofilm forming microorganisms isolated from chronic rhinosinusitis.

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Robert C Shields

Full Text Available BACKGROUND: The persistent colonization of paranasal sinus mucosa by microbial biofilms is a major factor in the pathogenesis of chronic rhinosinusitis (CRS. Control of microorganisms within biofilms is hampered by the presence of viscous extracellular polymers of host or microbial origin, including nucleic acids. The aim of this study was to investigate the role of extracellular DNA in biofilm formation by bacteria associated with CRS. METHODS/PRINCIPAL FINDINGS: Obstructive mucin was collected from patients during functional endoscopic sinus surgery. Examination of the mucous by transmission electron microscopy revealed an acellular matrix punctuated occasionally with host cells in varying states of degradation. Bacteria were observed in biofilms on mucosal biopsies, and between two and six different species were isolated from each of 20 different patient samples. In total, 16 different bacterial genera were isolated, of which the most commonly identified organisms were coagulase-negative staphylococci, Staphylococcus aureus and α-haemolytic streptococci. Twenty-four fresh clinical isolates were selected for investigation of biofilm formation in vitro using a microplate model system. Biofilms formed by 14 strains, including all 9 extracellular nuclease-producing bacteria, were significantly disrupted by treatment with a novel bacterial deoxyribonuclease, NucB, isolated from a marine strain of Bacillus licheniformis. Extracellular biofilm matrix was observed in untreated samples but not in those treated with NucB and extracellular DNA was purified from in vitro biofilms. CONCLUSION/SIGNIFICANCE: Our data demonstrate that bacteria associated with CRS form robust biofilms which can be reduced by treatment with matrix-degrading enzymes such as NucB. The dispersal of bacterial biofilms with NucB may offer an additional therapeutic target for CRS sufferers.

Time course of radiometric detection of positive blood cultures in childhood

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Meadow, W.L.; Schwartz, I.K.

1986-01-01

We have determined the time course of radiometric detection of microbial growth in 2348 positive blood culture specimens obtained at Wyler Children's Hospital during a 5-year interval. Overall 72 and 88% of isolates were detected within 48 and 72 hours after sampling, respectively. For pathogenic organisms aerobic detection was generally more rapid and more inclusive than anaerobic detection. At 48 hours of incubation the detection of six potential pathogens (Salmonella sp., Haemophilus influenzae, Group D streptococci, Neisseria meningitidis, coagulase-negative staphylococci, Candida sp.) was significantly delayed compared with detection of other pathogenic organisms recovered from blood. At 72 hours of incubation the detection rates remained less than 95% for H. influenzae, Staphylococcus aureus, Klebsiella sp., coagulase-negative staphylococci, Group D streptococci and Candida sp. These data should assist clinical decisions regarding duration of antibiotic therapy for the presumptive diagnosis of bacteremia in children

Time course of radiometric detection of positive blood cultures in childhood

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Meadow, W.L.; Schwartz, I.K.

1986-05-01

We have determined the time course of radiometric detection of microbial growth in 2348 positive blood culture specimens obtained at Wyler Children's Hospital during a 5-year interval. Overall 72 and 88% of isolates were detected within 48 and 72 hours after sampling, respectively. For pathogenic organisms aerobic detection was generally more rapid and more inclusive than anaerobic detection. At 48 hours of incubation the detection of six potential pathogens (Salmonella sp., Haemophilus influenzae, Group D streptococci, Neisseria meningitidis, coagulase-negative staphylococci, Candida sp.) was significantly delayed compared with detection of other pathogenic organisms recovered from blood. At 72 hours of incubation the detection rates remained less than 95% for H. influenzae, Staphylococcus aureus, Klebsiella sp., coagulase-negative staphylococci, Group D streptococci and Candida sp. These data should assist clinical decisions regarding duration of antibiotic therapy for the presumptive diagnosis of bacteremia in children.

Understanding Bacterial Isolates in Blood Culture and Approaches Used to Define Bacteria as Contaminants: A Literature Review.

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Hossain, Belal; Islam, Mohammad Shahidul; Rahman, Atiqur; Marzan, Mahfuza; Rafiqullah, Iftekhar; Connor, Nicholas E; Hasanuzzaman, Mohammad; Islam, Maksuda; Hamer, Davidson H; Hibberd, Patricia L; Saha, Samir K

2016-05-01

Interpretation of blood culture isolates is challenging due to a lack of standard methodologies for identifying contaminants. This problem becomes more complex when the specimens are from sick young infants, as a wide range of bacteria can cause illness among this group. We used 43 key words to find articles published between 1970 and 2011 on blood culture isolates and possible contaminants in the PubMed database. Experts were also consulted to obtain other relevant articles. Selection of articles followed systematic methods considering opinions from more than 1 reviewer. After reviewing the titles of 3869 articles extracted from the database, we found 307 relevant to our objective. Based on the abstracts, 42 articles were selected for the literature review. In addition, we included 7 more articles based on cross-references and expert advice. The most common methods for differentiating blood culture isolates were multiple blood cultures from the same subject, antibiograms and molecular testing. Streptococcus pneumoniae, Hemophilus influenzae, Neisseria meningitidis and group A and B streptococcus were always considered as pathogens, whereas Bacillus sp., Diphtheroids, Propionibacterium and Micrococcus were commonly regarded as contaminants. Coagulase-negative staphylococci were the most frequent isolates and usually reported as contaminants unless the patient had a specific condition, such as long-term hospitalization or use of invasive devices (catheters). Inaccurate interpretation of blood culture may falsely guide treatment and also has long-term policy implications. The combination of clinical and microbiological knowledge, patient's clinical history and laboratory findings are essential for appropriate interpretation of blood culture.

Resistance to Antibiotics in Strains of Staphylococcus spp., Enterococcus spp. and Escherichia coli Isolated from Rectal Swabs of Pigs

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M. Kolář

2008-01-01

Full Text Available The study aimed at determining the level of resistance of selected bacterial species (Staphylococcus spp., Enterococcus spp., Escherichia coli isolated from rectal swabs of pigs to antimicrobial agents. The tested strains were isolated from piglets aged 7 to 30 days. Bacterial species were identified by standard microbiological techniques and susceptibility to antibiotics was determined quantitatively by the standard microdilution method. Resistance of the Staphylococcus aureus strain to oxacillin was confirmed by detection of the mecA gene and PBP2a. A total of 115 Staphylococcus spp. isolates were collected. In the case of Staphylococcus aureus, the methicillin-resistant strain (MRSA was identified. Moreover, higher frequency of coagulase-negative staphylococci with minimum inhibitory concentration of oxacillin ≥ 0.5 mg/l was noticed. Inducible resistance to clindamycin in the Staphylococcus hominis strain was also detected. The strains of Enterococcus spp. (61 isolates exhibited high resistance to tetracycline (98.5%, erythromycin (86.8% and chloramphenicol (54.4%. Vancomycin-resistant enterococci were not isolated. In the case of Escherichia coli strains (111 isolates, higher frequency of resistant strains to tetracycline (81.1% and ampicillin (62.2% was documented. Resistance to fluoroquinolones and production of broad-spectrum β-lactamases was not noticed. The presented study may be considered as a pilot project assessing the prevalence of resistant bacteria in piglets kept on a single farm. It demonstrated the presence of resistant strains of Staphylococcus spp., including one MRSA strain, Enterococcus spp. and Escherichia coli. These strains may be present as a result of postnatal colonization with both bacterial microflora of dams and environmental microflora.

Conjunctival endogenous microbiota in patients submitted to cataract surgery Microbiota endógena conjuntival em pacientes submetidos à cirurgia de catarata

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Claudete I. Locatelli

2003-07-01

Full Text Available Bacterial isolation, identification and antimicrobial susceptibility tests were carried out in ocular material collected with swab and polimethylmethacrylate (PMMA or silicone intraocular lenses (IOL from forty six patients submitted to cataract surgery. Seventy six isolates and seven different microorganisms were identified. Coagulase-negative staphylococci (CNS were the predominant microorganisms isolated from swabs (71.4% of cases, PMMA lenses (81.3% and silicon lenses (77.8%. Coagulase-negative staphylococci isolates revealed high resistance to penicillin G followed by tetracycline, chloramphenicol and aminoglicosides. However, these isolates displayed great susceptibility to vancomycin, cephalothin and ofloxacin. Except for penicillin G, Staphylococcus aureus was very sensitive to the antimicrobial agents including oxacillin. Among Gram-negatives, Proteus mirabilis was prevalent and presented high resistance to tetracycline and chloramphenicol. Enterococcus isolates were vancomycin sensitive.A partir de material ocular coletado de 46 pacientes submetidos à cirurgia de catarata foram realizados isolamento, identificação e teste de susceptibilidade de microrganismos frente a antimicrobianos, utilizando-se suabes e lentes intraoculares (LIO. Foram obtidos 76 isolados e identificados 7 tipos de microrganismos. Estafilococos coagulase-negativos (CNS foram os microrganismos mais freqüentemente detectados de suabes (71,4% dos casos, lentes de PMMA (81,3% e lentes de silicone (77,8%. Isolados de CNS apresentaram elevada resistência à penicilina G, seguida por tetraciclina, cloranfenicol e aminoglicosídeos. No entanto, estes isolados mostraram grande sensibilidade à vancomicina, cefalotina e ofloxacina. Com exceção da penicilina G, os isolados de Staphylococcus aureus foram bastante sensíveis aos agentes antimicrobianos, incluindo a oxacilina. Entre as gram-negativas, Proteus mirabilis foi a bactéria mais freqüente e também se mostrou

Antistaphylococcal activity of LBM415, a new peptide deformylase inhibitor, compared with those of other agents.

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Credito, Kim; Lin, Gengrong; Ednie, Lois M; Appelbaum, Peter C

2004-10-01

The MICs of LBM415, a new peptide diformylase inhibitor, were isolates of Staphylococcus aureus and coagulase-negative staphylococci. LBM415 MICs were similar irrespective of whether the strains were methicillin susceptible or resistant. All strains were also susceptible to vancomycin, linezolid, ranbezolid, daptomycin, oritavancin, and quinupristin-dalfopristin. LBM415 at the MIC was bacteriostatic after 24 h.

[Identification of anaerobic gram-negative bacilli isolated from various clinical specimens and determination of antibiotic resistance profiles with E-test methods].

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Demir, Cengiz; Keşli, Recep

2018-01-01

The aim of this study was to identify gram-negative anaerobic bacilli isolated from various clinical specimens that were obtained from patients with suspected anaerobic infections and to determine the antibiotic resistance profiles by using the antibiotic concentration gradient method. The study was performed in Afyon Kocatepe University Ahmet Necdet Sezer Research and Practice Hospital, Medical Microbiology Laboratory between 1 November 2014 and 30 October 2015. Two hundred and seventyeight clinical specimens accepted for anaerobic culture were enrolled in the study. All the samples were cultivated anaerobically by using Schaedler agar with 5% defibrinated sheep blood and Schaedler broth. The isolated anaerobic gram-negative bacilli were identified by using both the conventional methods and automated identification system (VITEK 2, bioMerieux, France). Antibiotic susceptibility tests were performed with antibiotic concentration gradient method (E-test, bioMerieux, France); against penicillin G, clindamycin, cefoxitin, metronidazole, moxifloxacin, imipenem, meropenem, ertapenem and doripenem for each isolate. Of the 28 isolated anaerobic gram-negative bacilli; 14 were identified as Bacteroides fragilis group, 9 were Prevotella spp., and 5 were Fusobacterium spp. The highest resistance rate was found against penicillin (78.5%) and resistance rates against clindamycin and cefoxitin were found as 17.8% and 21.4%, respectively. No resistance was found against metronidazole, moxifloxacin, imipenem, meropenem, ertapenem and doripenem. As a result, isolation and identification of anaerobic bacteria are difficult, time-consuming and more expensive when compared with the cost of aerobic culture. The rate of anaerobic bacteria isolation may be increased by obtaining the appropriate clinical specimen and appropriate transportation of these specimens. We believe that the data obtained from the study in our center may offer benefits for the follow up and treatment of infections

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

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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. PMID:27482361

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

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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Karlowsky, James A; Hoban, Daryl J; Hackel, Meredith A; Lob, Sibylle H; Sahm, Daniel F

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. Copyright © 2017 Sociedade Brasileira de Infectologia. Published by Elsevier Editora Ltda. All rights reserved.

Microbiological Findings in Ready-to-Eat and Precooked Food Distributed in Public Catering Halls in Cagliari Province, Italy.

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Brignardello, Silvana; Sabiu, Rosangela; Tedde, Tiziana; Cocco, Enrica; Pitzalis, Gabriella; Meli, Clara; Cogoni, Maria Paola

2014-08-28

During this research 159 samples of ready-to-eat and precooked food were examined for the detection of Salmonella spp., Listeria monocytogenes , Escherichia coli , coagulase-positive staphylococci and Enterobacteriaceae , bacterial count. All samples were negative for Salmonella spp. and Listeria monocytogenes ; although a low count of coagulase-positive staphylococci (no. 5 samples) and E. coli (no. 3) was found. We proceeded with the identification of the isolates on every sample with high, moderate and low-count of Enterobacteriaceae , to better understand the microbial ecology. The Enterobacteriaceae species most frequently detected were Pantoea spp. (29.62%), Enterobacter cloacae (20.37%) and Serratia liquefaciens (12.96%); other different species of Enterobacteriaceae were detected with a ready-to-eat and precooked food, to proceed with the identification of every microorganism.

[A case of subarachnoid hemorrhage due to infective endocarditis by methicillin-resistant coagulase-negative staphylococcus].

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Kajikawa, Shunsuke; Oeda, Tomoko; Park, Kwiyoung; Yamamoto, Kenji; Sugiyama, Hiroshi; Sawada, Hideyuki

2017-12-27

A 77-year-old man visited our hospital with unstable gait following 2 months of anorexia. Brain MRI showed multiple infarcts; cardiac echocardiography revealed mitral-valve vegetation; and blood culture revealed methicillin-resistant coagulase-negative staphylococci. The patient was diagnosed with infective endocarditis (IE). Subarachnoid hemorrhage (SAH) developed ten days after antibiotic treatment. Intracranial aneurysm was not found. We speculated that chronic inflammation of the cerebral arterial walls by bacteria of low virulence was associated with SAH complication. The vegetation disappeared following additional gentamicin administration and the patient recovered to walk.

Detection and Antibiotic Susceptibility Pattern of Biofilm Producing Gram Positive and Gram Negative Bacteria Isolated From a Tertiary Care Hospital of Pakistan

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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.

Isolation, Biochemical and Molecular Identification, and In-Vitro Antimicrobial Resistance Patterns of Bacteria Isolated from Bubaline Subclinical Mastitis in South India.

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P L Preethirani

Full Text Available Buffaloes are the second largest source of milk. Mastitis is a major impediment for milk production, but not much information is available about bubaline mastitis, especially subclinical mastitis. The aim of this study was to (a investigate the application of various tests for the diagnosis of bubaline subclinical mastitis, (b identify the major bacteria associated with it, and (c evaluate the antibiotic resistance pattern of the bacteria. To this end, 190 quarter milk samples were collected from 57 domesticated dairy buffaloes from organized (64 samples and unorganized (126 samples sectors. Of these, 48.4%, 40.0%, 45.8%, 61.1%, and 61.6% were positive for subclinical mastitis by somatic cell count, electrical conductivity, California mastitis test, bromothymol blue test, and N-acetyl glucosaminidase test, respectively. As compared to the gold standard of somatic cell count, California mastitis test performed the best. However, a combination of the two methods was found to be the best option. Microbiological evaluation, both by biochemical methods as well as by monoplex and multiplex polymerase chain reaction, revealed that coagulase-negative staphylococci were the most predominant (64.8% bacteria, followed by streptococci (18.1%, Escherichia coli (9.8% and Staphylococcus aureus (7.3%. Most of the pathogens were resistant to multiple antibiotics, especially to β-lactam antibiotics. We propose that California mastitis test be combined with somatic cell count for diagnosis of subclinical mastitis in domestic dairy buffaloes. Further, our results reveal high resistance of the associated bacteria to the β-lactam class of antibiotics, and a possible major role of coagulase-negative staphylococci in causing the disease in India.

Isolation, Biochemical and Molecular Identification, and In-Vitro Antimicrobial Resistance Patterns of Bacteria Isolated from Bubaline Subclinical Mastitis in South India.

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Preethirani, P L; Isloor, Shrikrishna; Sundareshan, S; Nuthanalakshmi, V; Deepthikiran, K; Sinha, Akhauri Y; Rathnamma, D; Nithin Prabhu, K; Sharada, R; Mukkur, Trilochan K; Hegde, Nagendra R

2015-01-01

Buffaloes are the second largest source of milk. Mastitis is a major impediment for milk production, but not much information is available about bubaline mastitis, especially subclinical mastitis. The aim of this study was to (a) investigate the application of various tests for the diagnosis of bubaline subclinical mastitis, (b) identify the major bacteria associated with it, and (c) evaluate the antibiotic resistance pattern of the bacteria. To this end, 190 quarter milk samples were collected from 57 domesticated dairy buffaloes from organized (64 samples) and unorganized (126 samples) sectors. Of these, 48.4%, 40.0%, 45.8%, 61.1%, and 61.6% were positive for subclinical mastitis by somatic cell count, electrical conductivity, California mastitis test, bromothymol blue test, and N-acetyl glucosaminidase test, respectively. As compared to the gold standard of somatic cell count, California mastitis test performed the best. However, a combination of the two methods was found to be the best option. Microbiological evaluation, both by biochemical methods as well as by monoplex and multiplex polymerase chain reaction, revealed that coagulase-negative staphylococci were the most predominant (64.8%) bacteria, followed by streptococci (18.1%), Escherichia coli (9.8%) and Staphylococcus aureus (7.3%). Most of the pathogens were resistant to multiple antibiotics, especially to β-lactam antibiotics. We propose that California mastitis test be combined with somatic cell count for diagnosis of subclinical mastitis in domestic dairy buffaloes. Further, our results reveal high resistance of the associated bacteria to the β-lactam class of antibiotics, and a possible major role of coagulase-negative staphylococci in causing the disease in India.

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