Heilmann, Christine; Peters, Georg
SUMMARY The definition of the heterogeneous group of coagulase-negative staphylococci (CoNS) is still based on diagnostic procedures that fulfill the clinical need to differentiate between Staphylococcus aureus and those staphylococci classified historically as being less or nonpathogenic. Due to patient- and procedure-related changes, CoNS now represent one of the major nosocomial pathogens, with S. epidermidis and S. haemolyticus being the most significant species. They account substantially for foreign body-related infections and infections in preterm newborns. While S. saprophyticus has been associated with acute urethritis, S. lugdunensis has a unique status, in some aspects resembling S. aureus in causing infectious endocarditis. In addition to CoNS found as food-associated saprophytes, many other CoNS species colonize the skin and mucous membranes of humans and animals and are less frequently involved in clinically manifested infections. This blurred gradation in terms of pathogenicity is reflected by species- and strain-specific virulence factors and the development of different host-defending strategies. Clearly, CoNS possess fewer virulence properties than S. aureus, with a respectively different disease spectrum. In this regard, host susceptibility is much more important. Therapeutically, CoNS are challenging due to the large proportion of methicillin-resistant strains and increasing numbers of isolates with less susceptibility to glycopeptides. PMID:25278577
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
Kudinha, T.; C. Simango
This study was carried out to determine the prevalence of coagulase-negative staphylococci in clinical and subclinical mastitis in commercial and small-scale farms in Zimbabwe. Thirty five quarter milk samples from clinical mastitis cases and 371 quarter milk samples from cows with subclinical mastitis were cultured for bacterial pathogens. The most frequent pathogens isolated in clinical mastitis were the enteric bacteria (31.4 %), followed by coagulase negative staphylococci (22.9 %) and th...
Kudinha, T; Simango, C
This study was carried out to determine the prevalence of coagulase-negative staphylococci in clinical and subclinical mastitis in commercial and small-scale farms in Zimbabwe. Thirty five quarter milk samples from clinical mastitis cases and 371 quarter milk samples from cows with subclinical mastitis were cultured for bacterial pathogens. The most frequent pathogens isolated in clinical mastitis were the enteric bacteria (31.4%), followed by coagulase negative staphylococci (22.9%) and then Staphylococcus aureus (17.1%), whereas in subclinical mastitis S. aureus (34.2%) and coagulase-negative staphylococci were (33.2%) the most common. Bacillus species were only isolated in milk samples from subclinical mastitis. Coagulase-negative staphylococci were observed in mixed infections with other bacteria in only 2.2 of the 406 milk samples from clinical and subclinical mastitis where they were isolated together with Bacillus species in 6 of the 9 mixed infection cases. About 95% of the milk samples from which 131 coagulase-negative staphylococci were isolated had correspondingly high somatic cell counts. The coagulase-negative staphylococci isolated most frequently were S. chromogenes (7.9%), S. epidermidis (7.4%) and S. hominis (5.9%). They were all associated with high somatic cell counts. All the coagulase-negative staphylococci isolates were susceptible to cloxacillin and erythromycin, and more than 90% of the isolates were susceptible to neomycin, penicillin and streptomycin. The highest resistance was to tetracycline (17.6%), followed by lincomycin (13.7%). About 8% of the isolates were resistant to both penicillin and streptomycin.
Full Text Available This study was carried out to determine the prevalence of coagulase-negative staphylococci in clinical and subclinical mastitis in commercial and small-scale farms in Zimbabwe. Thirty five quarter milk samples from clinical mastitis cases and 371 quarter milk samples from cows with subclinical mastitis were cultured for bacterial pathogens. The most frequent pathogens isolated in clinical mastitis were the enteric bacteria (31.4 %, followed by coagulase negative staphylococci (22.9 % and then Staphylococcus aureus (17.1 %, whereas in subclinical mastitis S. aureus (34.2 % and coagulase-negative staphylococci were (33.2 % the most common. Bacillus species were only isolated in milk samples from subclinical mastitis. Coagulase-negative staphylococci were observed in mixed infections with other bacteria in only 2.2 % of the 406 milk samples from clinical and subclinical mastitis where they were isolated together with Bacillus species in 6 of the 9 mixed infection cases. About 95 % of the milk samples from which 131 coagulase-negative staphylococci were isolated had correspondingly high somatic cell counts. The coagulase-negative staphylococci isolated most frequently were S. chromogenes (7.9 %, S. epidermidis (7.4 % and S. hominis (5.9 %. They were all associated with high somatic cell counts. All the coagulase-negative staphylococci isolates were susceptible to cloxacillin and erythromycin, and more than 90 %of the isolates were susceptible to neomycin, penicillin and streptomycin. The highest resistance was to tetracycline (17.6 %, followed by lincomycin (13.7 %. About 8 % of the isolates were resistant to both penicillin and streptomycin.
Full Text Available The study was carried out to investigate the prevalence and antimicrobial susceptibility of Coagulase-Negative Staphylococci isolated from Bovine Mastitis in and around Dharwad region. A total of 310 samples were screened and 180 confirmed Coagulase-Negative Staphylococci were obtained. The antimicrobial susceptibility of Coagulase-Negative Staphylococci against 10 antimicrobial agents was tested using the disc diffusion method. The highest numbers of Coagulase-Negative Staphylococci were susceptible to ceftriaxone 83.88% followed by cefotaxime 79.41%, methicillin 76.47%, ciprofloxacin 73.52%, erythromycin 70.05%, amikacin 66.11%, gentamycin 42.94%, amoxicillin 36.76%, ampicillin 29.41%, and the lowest susceptibility was shown in penicillin 23.23% . The results indicated that the increase in prevalence and antibiotic resistance pattern of the Coagulase-Negative Staphylococci isolated from bovine mastitis exhibited the highest degree of susceptible to ceftriaxone of all the tested antimicrobial agents. [Vet. World 2011; 4(4.000: 158-161
de Freitas Guimarães, Felipe; Nóbrega, Diego Borin; Richini-Pereira, Virginia Bodelão; Marson, Pâmela Merlo; de Figueiredo Pantoja, José Carlos; Langoni, Helio
The objective of this study was to isolate and identify the main staphylococcal species causing bovine mastitis in 10 Brazilian dairy herds and study their capability to produce enterotoxins. Herds were selected based on size and use of milking technology, and farms were visited once during the study. All mammary glands of all lactating cows were screened using the California Mastitis Test (CMT) and a strip cup. A single aseptic milk sample (20 mL) was collected from all CMT-positive quarters. Identification of Staphylococcus spp. was performed using conventional microbiology, and PCR was used to determine the presence of enterotoxin-encoding genes (sea, seb, sec, and sed). Of the 1,318 CMT-positive milk samples, Staphylococcus spp. were isolated from 263 (19.9%). Of these isolates, 135 (51%) were coagulase-positive staphylococci (CPS) and 128 (49%) were coagulase-negative staphylococci (CNS). Eighteen different species of CNS were isolated, among which S. warneri, S. epidermidis and S. hyicus were the most frequent. The distribution of Staphylococcus species was different among herds: S. epidermidis was found in 8 herds, S. warneri was found in 7 herds, and S. hyicus in 6 herds. Some of the CNS species (S. saprophyticus ssp. saprophyticus, S. auricularis, S. capitis, and S. chromogenes) were isolated in only one of the farms. Genes related to production of enterotoxins were found in 66% (n=85) of all CNS and in 35% of the CPS isolates. For both CNS and CPS isolates, the most frequently identified enterotoxin genes were sea, seb, and sec; the prevalence of sea differed between CPS (9.5%) and CNS (35.1%) isolates. Staphylococcus warneri isolates showed a greater percentage of sea than seb, sec, or sed, whereas S. hyicus isolates showed a greater percentage of sea than sec. Over 60% of CNS belonged to 3 major species, which carried 62.2 to 81.3% of the enterotoxin genes. The high prevalence highlights the potential for food poisoning caused by these species. For
Xu, Jiancheng; Yan, Chaoying; Wang, Liqiang; Zhou, Qi
This study was to investigate the antimicrobial resistance of coagulase-negative Staphylococci isolated in 8 consecutive years in the First Bethune Hospital. Disk diffusion test was used to study the antimicrobial resistance. The data were analyzed by WHONET 5 software according to Clinical and Laboratory Standards Institute (CLSI). Most of 2484 strains of coagulase-negative Staphylococci were collected from blood 925 (37.2%), secretions 652 (26.2%) and urine 323 (13.0%) during the past 8 years. The rates of methicillin-resistant coagulase-negative Staphylococci (MRCNS) were between 79.4% and 81.5% during the past 8 years, respectively. In recent 8 years, the antimicrobial resistance of coagulase-negative Staphylococci had increased. Monitoring the antimicrobial resistance to coagulase-negatives Staphylococci should be strengthened. The change of the antimicrobial resistance should be investigated in order to direct rational drug usage in the clinic and prevent bacterial strain of drug resistance from being transmitted.
Full Text Available Introduction: Drug resistant Staphylococci are the most important agents of nosocomial infections. In this survey, effect of different antibiotics on these bacteria and their drug resistance was investigated. Methods: The study included 500 strains of Staphylococci. Minimum Inhibitory Concentrations of all antibiotics was determined by the broth macro dilution technique and standard methods from the National Committee for Clinical Laboratory Standards. Result : Oxacillin resistance of S. aureus was 14.2% and that of coagulase-negative staphylococci was 53.4%. The activity of different antibiotics is presented in detail. Conclusion: Surveillance of strains resistant to methicillin is necessary.
Full Text Available Resistance of staphylococci to methicillin is important especially in the case of Staphylococcus aureus isolates. Its impact in veterinary medicine is not exactly specified in coagulase-negative staphylococci; however, these staphylococci may represent an important reservoir of resistance genes. The study aimed at detecting resistance to methicillin in coagulase-negative staphylococci from raw materials and foodstuffs of animal origin and assessing the tests frequently used to determine this resistance. Coagulase-negative staphylococci (198 isolates of 12 species were tested. Resistance to methicillin was determined by the disk diffusion method using oxacillin and cefoxitin disks, microdilution method, detection of PBP2a and the mecA gene. Of the tested isolates, 109 (55.1% were classified as resistant by the diffusion test with oxacillin, 32 isolates (16.2% by the test with cefoxitin and 50 isolates (25.3% on the basis of oxacillin minimum inhibitory concentration (MIC. No resistant isolates were incorrectly identified as susceptible when using the disk diffusion method with oxacillin (sensitivity of 100%. However, apart from 22 correctly classified resistant isolates, another 87 isolates were incorrectly identified as resistant as well (specificity of 50.6%. The test with cefoxitin showed the lowest (45.5% sensitivity in determination of resistant isolates. By contrast, this test was the most precise in classification of resistant isolates (specificity of 87.5%. When using the microdilution method, resistant strains were identified with the sensitivity and specificity of 68.2% and 80.1%, respectively. The results revealed substantial variability of methicillin-resistant isolates ranging from 16.2% to 55.1%, depending on the phenotyping methods and recommended interpretation criteria used. Therefore, it is advisable to reconsider the current interpretation criteria in the case of coagulasenegative staphylococci of animal origin (with the
Full Text Available In this research, the susceptibility to clindamycin, tetracycline, amikacin, amoxicillin + clavulanic acid, enrofloxacine, vancomycin, trimethoprim + sulphametoxazol, tobramycin, chloramphenicol, ciprofloxacin, erythromycin, penicillin and trimethoprim was tested in coagulase-negative staphylococci (n=78 and lactic acid bacteria (n=30 by means of disk diffusion test and E-test. The isolates were collected from soft and hard cheeses, butter and brine. All isolates of coagulase-negative staphylococci were susceptible to clindamycin, amikacin, amoxicillin + clavulanic acid, enrofloxacine, vancomycin, chloramphenicol and ciprofloxacin according to CLSI breakpoints. A total of 30 staphylococci isolates (38.46 % were resistant to erythromycin, 18 to penicillin (23.07 %, 4 to tetracycline (5.12 %, and one isolate to trimethoprim, tobramicin and trimethoprim + sulphametoxazol (1.28 %. Among 78 tested staphylococci, 35 of them were resistant to at least one antimicrobial substance (44.87 %. The rate of resistant isolates of different soft cheese types ranged from 22 to 70 %, while resistant staphylococci were absent in hard cheese and brine. The growth of lactic acid bacteria was not influenced by trimethoprim + sulphametoxazol (n=29, vancomycin (n=29, trimethoprim (n=28, amikacin (n=10 and tobramycin (n=10. The results show that significant part of apathogenic microbiota in different dairy products is phenotypically resistant to antimicrobial agents.
Fowoyo, P T; Ogunbanwo, S T
Coagulase-negative staphylococci have become increasingly recognized as the etiological agent of some infections. A significant characteristic of coagulase-negative staphylococci especially strains isolated from animals and clinical samples is their resistance to routinely used antibiotics although, resistant strains isolated from fermented foods have not been fully reported. A total of two hundred and fifty-five CoNS isolates were subjected to antimicrobial susceptibility test using the disc diffusion technique. The minimum inhibitory concentration of the isolates to the tested antibiotics was determined using the microbroth dilution method. Methicillin resistant strains were confirmed by detection of methicillin resistant genes (mecA) and also employing cefoxitin screening test. The isolates were confirmed to be methicillin resistant by the detection of mecA genes and the cefoxitin screening test. The isolates demonstrated appreciable resistance to ampicillin (86.7%), sulfomethoxazole-trimethoprim (74.9%), amoxicillin-clavulanic acid (52.5%) and oxacillin (35.7%). Methicillin resistance was exhibited by 13 out of the 255 isolates although no mecA gene was detected. It was also observed that the methicillin resistant isolates were prevalent in these traditional foods; iru, kindirmo, nono and wara. This study has ameliorated the incidence of multiple antibiotic resistant coagulase-negative staphylococci in Nigerian fermented foods and if not tackled adequately might lead to horizontal transfer of antibiotic resistance from food to man.
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
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.
Sampimon, O C; Vernooij, J C A; Mevius, D J; Sol, J
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 coagulase-negative staphylococci. In 2004 this proportion was 42.2%. The proportion of coagulase-negative staphylococci of the bacteria isolated from milk samples from cows with clinical mastitis was 7.3% in 1999 and 14.1% in 2004. In this study, the susceptibility of 108 coagulase-negative staphylococci to oxacillin, cefquinome, streptomycin, neomycin, penicillin, and the combination of nafcillin, penicillin, and streptomycin was tested. The isolates were cultured from milk collected from cows with mastitis and typed using the Api-Staph system. Eight species were identified. Staphylococcus chromogenes was the predominant species (41.7%), followed by Staphylococcus xylosus (15.7%) and Staphylococcus simulans (10.2%). With the agar dilution method all strains proved to be sensitive to cefquinome and 90% to oxacillin. Three isolates (2.8%) were mecA-positive. Despite the agar dilution results, these three isolates should be considered resistant to all beta-lactam antibiotics (penicillins, penicillins combined with a beta-lactamase inhibitor and all generations of cephalosporins). In the agar diffusion test, all isolates proved to be sensitive to the combination of nafcillin-penicillin-streptomycin, 99% were sensitive to neomycin and 1% intermediate sensitive, and 95% were sensitive to streptomycin, 4% resistant, and 1% intermediate sensitive. The coagulase-negative staphylococci were highly resistant to penicillin (37.4%), although the level of resistance varied between species, from 0% for Staphylococcus simulans to 100% for Staphylococcus saprophyticus. Because coagulase-negative staphylococci are resistant to several antibiotics, sensitivity testing is important for targeted treatment of mastitis.
John, Joseph F.; Harvin, Alexander M
Coagulase-negative staphylococci (CNS) are a heterogenous group of Gram-positive cocci that are widespread commensals among mammalia. Unlike their coagulase-positive counterpart, Staphylococcus aureus, CNS produce few virulence patterns and normally refrain from invading tissue. Yet, not only can CNS cause infections in normal host tissue, but modern medicine has also seen their rise as opportunists that display adherence to medical device materials to produce a protective biofilm. CNS have h...
Kennedy F. Chah
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.
Full Text Available Antimicrobial resistance patterns and gene coding for methicillin resistance (mecA were determined in 25 S. aureus and 75 Coagulase Negative Staphylococci (CNS strains isolates from half-udder milk samples collected from goats with subclinical mastitis. Fourteen (56.0% S. aureus and thirty-one (41.3% CNS isolates were resistant to one or more antimicrobial agents. S. aureus showed the highest resistance rate against kanamycin (28.0%, oxytetracycline (16.0%, and ampicillin (12.0%. The CNS tested were more frequently resistant to ampicillin (36.0% and kanamycin (6.7%. Multiple antimicrobial resistance was observed in eight isolates, and one Staphylococcus epidermidis was found to be resistant to six antibiotics. The mecA gene was not found in any of the tested isolates. Single resistance against β-lactamics or aminoglicosides is the most common trait observed while multiresistance is less frequent.
J. Sedef Göçmen
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
Maria de Lourdes RS Cunha
Full Text Available Coagulase-negative staphylococci (CNS species identification is still difficult for most clinical laboratories. The scheme proposed by Kloos and Schleifer and modified by Bannerman is the reference method used for the identification of staphylococcal species and subspecies; however, this method is relatively laborious for routine use since it requires the utilization of a large number of biochemical tests. The objective of the present study was to compare four methods, i.e., the reference method, the API Staph system (bioMérieux and two methods modified from the reference method in our laboratory (simplified method and disk method, in the identification of 100 CNS strains. Compared to the reference method, the simplified method and disk method correctly identified 100 and 99% of the CNS species, respectively, while this rate was 84% for the API Staph system. Inaccurate identification by the API Staph method was observed for Staphylococcus epidermidis (2.2%, S. hominis (25%, S. haemolyticus (37.5%, and S. warneri (47.1%. The simplified method using the simple identification scheme proposed in the present study was found to be efficient for all strains tested, with 100% sensitivity and specificity and proved to be available alternative for the identification of staphylococci, offering, higher reliability and lower cost than the currently available commercial systems. This method would be very useful in clinical microbiology laboratory, especially in places with limited resources.
Ciro C. Rossi
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.
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.
Schoenfelder, Sonja M K; Dong, Ying; Feßler, Andrea T; Schwarz, Stefan; Schoen, Christoph; Köck, Robin; Ziebuhr, Wilma
Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) have globally emerged in animal husbandry. In addition to methicillin resistance, LA-MRSA may carry a variety of novel and uncommon antimicrobial resistance genes. Occurrence of the same resistance genes in coagulase-negative staphylococci (CoNS) and S. aureus suggests an ongoing genetic exchange between LA-MRSA and other staphylococci whose driving forces in the ecological niche of the farm environment are, however, still poorly understood. To assess the potential of CoNS as putative reservoirs for antibiotic resistance genes, we analysed the antimicrobial susceptibility of CoNS from dust and manure samples obtained in 41 pig farms in Germany, most of them (36 of 41) with a proven LA-MRSA/MSSA history. Among the 344 isolates analysed, 18 different CoNS species were identified and S. sciuri represented the most prevalent species (46%). High resistance rates were detected for tetracycline (71%), penicillin (65%) and oxacillin (64%) as well as fusidic acid (50%), which was mainly due to reduced susceptibility among S. sciuri isolates. S. sciuri exhibited pronounced multiresistance, and many isolates were characterised by the carriage of a number of uncommon (multi)resistance genes (e.g. cfr, apmA, fexA) and decreased susceptibility towards last resort antibiotics such as linezolid and daptomycin. The combined data suggest that S. sciuri harbours a significant resistance gene pool that requires further attention. We hypothesise that members of this species, due to their flexible lifestyle, might contribute to the spread of such genes in livestock environments. Copyright © 2016 Elsevier B.V. All rights reserved.
Klimiene, I; Virgailis, M; Pavilonis, A; Siugzdiniene, R; Mockeliunas, R; Ruzauskas, M
The objectives of this study were to determine the prevalence and antimicrobial resistance of coagulase-negative staphylococci (CNS) isolated from dairy cows with subclinical mastitis. Antimicrobial resistance in staphylococci were evaluated by breakpoint values specific to the species (EU-CAST). The presence of resistance-encoding genes was detected by multiplex PCR. A total of 191 CNS isolates were obtained. The CNS isolates were typically resistant to penicillin (67.4%), tetracyc-line (18.9%), and erythromycin (13.7%). CNS isolates (78.0%) were resistant to at least one antimicrobial compound, and 22.0% were multiresistant. The multiresistant isolates were predominantly Staphylococcus chromogenes (28.6%), Staphylococcus warneri (19%) and Staphylococcus haemolyticus (14.3%). According to MIC pattern data, multiresistant isolates showed the highest resistance (p<0.05) rates to penicillin (85.7%), tetracycline (66.7%), and erythromycin (48.2%), but all of them were sensitive to daptomycin, oxacillin, qiunupristin/dalfopristin, and vancomycin. S. chromogenes (9.5%), S. haemolyticus (4.8%), and S. capitis ss capitis (2.4%) strains were resistant to methicillin; their resistance to oxacillin and penicillin was more than 8 mg/l. A high rate of resistance to penicillin was linked to a blaZ gene found in 66.6% of the isolated multiresistant CNS strains. Resistance to tetracycline via the tetK (38.1%) gene and penicillin via the mecA (23.8%) gene were detected less frequently. Gene msrAB was responsible for macrolides and lincosamides resistance and detected in 28.6% of the CNS isolates. Antimicrobial resistance genes were identified more frequently in S. epidermidis, S. chromogenes, and S. warneri.
H.F.M. Willemse-Erix; J-W. Jachtenberg (Jan-Willem); H. Barutçi; G.J. Puppels (Gerwin); A.F. van Belkum (Alex); A. Voss (Andreas); K. Maquelin (Kees)
textabstractCoagulase-negative staphylococci (CNS) are among the most frequently isolated bacterial species in clinical microbiology, and most CNS-related infections are hospital acquired. Distinguishing between these frequently multiple-antibiotic-resistant isolates is important for both treatment
Sánchez Mainar, María; Weckx, Stefan; Leroy, Frédéric
Within ecosystems that are poor in carbohydrates, alternative substrates such as arginine may be of importance to coagulase-negative staphylococci (CNS). However, the versatility of arginine conversion in CNS remains largely uncharted. Therefore, a set of 86 strains belonging to 17 CNS species was screened for arginine deiminase (ADI), arginase, and nitric oxide synthase (NOS) activities, in view of their ecological relevance. In fermented meats, for instance, ADI could improve bacterial comp...
Sousa, Margarida; Silva, Nuno; Igrejas, Gilberto; Sargo, Roberto; Benito, Daniel; Gómez, Paula; Lozano, Carmen; Manageiro, Vera; Torres, Carmen; Caniça, Manuela; Poeta, Patrícia
Wild animal populations in contact with antimicrobials and antimicrobial resistant bacteria that are daily released into the environment are able to become unintentional hosts of these resistant microorganisms. To clarify this issue, our study evaluated the presence of antibiotic resistance determinants on coagulase-negative staphylococci recovered from birds of prey and studied their genetic relatedness by pulsed-field gel electrophoresis (PFGE). The unusual vga(A) and erm(T) genes, which co...
Bansal, B. K.; Gupta, D. K.; T. A. Shafi; Sharma, S
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...
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.
Jakeen K. El-Jakee
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.
Zadoks, Ruth N; Watts, Jeffrey L
Coagulase-negative staphylococci (CNS) are isolated commonly from bovine milk and skin. Their impact on udder health and milk quality is debated. It has been suggested that sources and consequences of infection may differ between CNS species. Species-specific knowledge of the impact and epidemiology of CNS intramammary infections is necessary to evaluate whether species-specific infection control measures are feasible and economically justified. Accurate measurement of impact, sources, and transmission mechanisms requires accurate species level identification of CNS. Several phenotypic and genotypic methods for identification of CNS species are available. Many methods were developed for use in human medicine, and their ability to identify bovine CNS isolates varies. Typeability and accuracy of typing methods are affected by the distribution of CNS species and strains in different host species, and by the ability of test systems to incorporate information on new CNS species into their experimental design and reference database. Generally, typeability and accuracy of bovine CNS identification are higher for genotypic methods than for phenotypic methods. As reviewed in this paper, DNA sequence-based species identification of CNS is currently the most accurate species identification method available because it has the largest reference database, and because a universally meaningful quantitative measure of homology with known species is determined. Once sources, transmission mechanisms, and impact of different CNS species on cow health, productivity and milk quality have been identified through use of epidemiological data and accurate species identification methods, appropriate methods for routine use in research and diagnostic laboratories can be proposed.
Fowoyo, P T; Ogunbanwo, S T
The incidence of coagulase-negative staphylococci (CoNS) may render food unsafe, as the clinical isolates have been reported to exude virulent traits. A total of 255 CoNS isolates from 6 traditional fermented foods (nono, kunu, wara, iru, ogi, and kindirmo) from North Central Nigeria, identified as Staphylococcus epidermidis, Staphylococcus simulans, Staphylococcus xylosus, Staphylococcus kloosii, and Staphylococcus caprae, were investigated for virulence traits. The strains were examined for biofilm formation and production of hyaluronidase, DNase, TNase, haemolysins, and superantigenic toxins (SEA, SEB, SEC, SED, and TSST-1) using standard and genotypic methods. The analysis of virulence factors revealed the production of slime in 200 isolates (78.4%); α-haemolysin in 136 (53.3%); β-haemolysin in 43 (16.9%); DNase in 199 (78.0%); TNase in 29 (11.4%); hyaluronidase in 125 (49.0%); TSST-1 in 119 (46.7%); and enterotoxin-producing isolates SEA, SEB, SEC, and SED in 61 (23.9%), 19 (7.5%), 9 (3.5%), and 8 (3.1%), respectively. PCR analysis detected tsst-1, sea, seb, and sec genes. The ability of these microorganisms to exhibit virulence evokes the potential to cause disease especially under determinate conditions or in immune-compromised patients. The occurrence of CoNS in food should not be ignored nor their pathogenic potential considered as insignificant, rather safety measures should be taken to reduce or totally eliminate their occurrence in foods.
Full Text Available Ghotaslou R1, Gharashi Z2, Pour-asl M3 1. Assistant professor, Department of pathobiology, Faculty of medicine, Tabriz University of medical sciences 2. Associate professor, Department of pediatrics, Tabriz University of medical sciences 3. Instructor, Department of pathobiology, Faculty of health, Tabriz University of medical sciences Abstract Background: The most common microorganism in blood cultures is Coagulase Negative Staphylococci (CNSt. Exact diagnosis of bacterial agents in blood and their antibiograms are an important value in septicemia therapy. The purpose of the present study is to identify and determine antibiotic susceptibility of CNSt isolated from blood in children. Materials and methods: Samples were collected from 270 patients with CNSt positive blood cultures during 2 years (2003-2004 in pediatric center of Tabriz by routine microbiological methods. The disc agar diffusion technique (kriby-Bauer was used to evaluate the susceptibility test of CNSt. Results: In our study mean age of patients was 10 month. 54.4% and 45.6% were male and female repectively. In 270 CNSt, the resistance to Penicillin, Ampicilline, Cephalexin, Ceftizoxime, Ceftriaxon and Vancomycin were respectively 98.1%, 95.9%, 62.3%, 55.9%, 53.7% and 2.6%. Conclusion: Antibiotic susceptibility of CNSt is not predictable and multiresistant strains are common. Vancomycin is the drug of choice, since only 2.6% resistant CNSt were noticed.
Valeria Cataneli Pereira
Full Text Available Coagulase-negative staphylococci (CoNS are the microorganisms most frequently isolated from clinical samples and are commonly found in neonatal blood cultures. Oxacillin is an alternative treatment of choice for CoNS infections; however, resistance to oxacillin can have a substantial impact on healthcare by adversely affecting morbidity and mortality. The objective of this study was to detect and characterise oxacillin-resistant CoNS strains in blood cultures of newborns hospitalised at the neonatal ward of the University Hospital of the Faculty of Medicine of Botucatu. One hundred CoNS strains were isolated and the mecA gene was detected in 69 of the CoNS strains, including 73.2% of Staphylococcus epidermidis strains, 85.7% of Staphylococcus haemolyticus strains, 28.6% of Staphylococcus hominis strains and 50% of Staphylococcus lugdunensis strains. Among these oxacillin-resistant CoNS strains, staphylococcal cassette chromosome mec (SCCmec type I was identified in 24.6%, type II in 4.3%, type III in 56.5% and type IV in 14.5% of the strains. The data revealed an increase in the percentage of CoNS strains isolated from blood cultures from 1991-2009. Furthermore, a predominant SCCmec profile of the oxacillin-resistant CoNS strains isolated from neonatal intensive care units was identified with a prevalence of SCCmec types found in hospital-acquired strains.
Abimanyu, Nagarajan; Krishnan, Arunkumar; Murugesan, Saravanan; Subramanian G, Kaushik; Gurumurthy, Sivakumar; Krishnan, Padma
Introduction: Methicillin-Resistant Staphylococcus aureus (MRSA) has become a major public health problem in both hospitals and communities. Panton – Valentine Leucocidin (PVL) has been reported to be an important marker for the highly pathogenic community acquired S. aureus infections. A rapid detection of these MRSA is very important for its treatment. The specific detection of MRSA is always a problem due to the prevalence of methicillin resistance among the coagulase negative Staphylococci. Hence, this study was done to develop a rapid triplex PCR for the detection of PVL positive MRSA and for the simultaneous differentiation of MRSA from Coagulase Negative Staphylococci (CoNS). Materials and Methods: We developed a triplex PCR for the specific detection of PVL positive Community Acquired (CA) – MRSA and for its simultaneous differentiation from the coagulase negative Staphylococci. We used PCR for targeting the fem A gene which is specific for S. aureus, mecA which is specific for methicillin-resistance and luk - PV which is specific for the PVL toxin. The method was evaluated with a total of 100 clinical isolates of Staphylococcus spp. Results: The triplex PCR was successfully standardized by using the reference strains and it was evaluated by using clinical strains. The method was found to be rapid, highly sensitive (100%), specific (99%) and cost effective. Conclusion: Triplex PCR can be used as a diagnostic tool for the detection of the highly pathogenic strains of CA-MRSA. PMID:23542876
Full Text Available Aims: It is very important to detect the coagulase-negative Staphylococci, which produce the hospital infections. Being one of the most expensive and time-consuming stages before the polymerase chain reaction (PCR, DNA extraction is one of the primary stages of PCR. Then, it should be noticed that the elimination of the stage might save time and costs. The aim of this study was to compare two PCR methods including the method with the utilization of the extracted DNA with the extraction kit and the direct PCR method in the detection of the methicillin-resistant genes in the coagulase-negative Staphylococci. Materials & Methods: In the descriptive cross-sectional study, 135 Staphylococcus epidermidis and 88 Staphylococcus saprophyticus samples were studied, separated from blood, wounds, urinary catheter, and urine samples of patients hospitalized in the treatment centers of Zahedan. The direct PCR was done on the Staphylococcus saprophyticus and Staphylococcus epidermidis colonies. PCR with the extracted DNA was done for mecA and 16srDNA genes using the extraction kit, and the results were compared. Findings: In both methods, mecA and 16srDNA genes were successfully amplified in 310bp and 420bp related to Staphylococcus bacteria identifying gene and methicillin resistant gene, respectively. In addition, there were approximately the same band qualities. Conclusion: In order to save time and costs, the direct PCR method can be used to detect methicillin-resistant coagulase-negative Staphylococci.
Hogt, A.H.; Dankert, J.; Feijen, Jan
Adhesion of coagulasef-negative staphylococci (CNS) was studied onto a homologous series of methacrylate polymers and copolymers. The materials varied in wettability (contact angles) and were either positively or negatively charged (zetapotential). Bacterial adhesion experiments performed in a
Goldstein, Rachel Rosenberg; Kleinfelter, Lara; He, Xin; Micallef, Shirley A; George, Ashish; Gibbs, Shawn G; Sapkota, Amy R
Coagulase negative staphylococci (CoNS) are leading causes of nosocomial infections and community-acquired methicillin-resistant CoNS (MRCoNS) infections are increasing. CoNS have been previously detected in reclaimed water. To date, no studies have evaluated the prevalence of CoNS carriage among humans exposed to reclaimed water in the U.S. We examined the prevalence and odds of CoNS and antibiotic-resistant CoNS carriage in spray irrigators exposed to reclaimed water compared to controls. We collected nasal and dermal swab samples from 19 reclaimed water spray irrigation workers (n=96 total samples) and 24 controls (n=92 total samples). Samples were analyzed for CoNS using culture-based assays. Isolates were confirmed using biochemical tests and PCR. Antimicrobial susceptibility testing was performed using disk diffusion. Data were analyzed by two-sample proportion tests, logistic regression, and generalized linear mixed effects models. The prevalence of CoNS, antibiotic-resistant CoNS, and MRCoNS carriage among spray irrigation workers was 79% (15/19), 32% (6/19), and 16% (3/19), compared to 13% (3/24), 4% (1/24), and 0% (0/24) of controls. Spray irrigators were more likely to be carriers of CoNS (p<0.01), antibiotic-resistant CoNS (p<0.01), and MRCoNS (p=0.02) compared to controls. The odds of CoNS carriage significantly increased with exposure to reclaimed water (p=0.04) even accounting for changes over time (p=0.05). Our data highlight the need to further examine the potential dissemination of CoNS and antibiotic-resistant CoNS from reclaimed water into the environment and human communities and related public health implications. Copyright © 2017 Elsevier B.V. All rights reserved.
Asaad, Ahmed Morad; Ansar Qureshi, Mohamed; Mujeeb Hasan, Syed
Identification of coagulase-negative staphylococci (CoNS) as nosocomial pathogens or contaminants is significant for microbiologists and clinicians. This study aimed to determine the frequency of isolation and antimicrobial resistance patterns of CoNS isolates from nosocomial bloodstream infections (BSIs) and to identify risk factors associated with true bacteremia caused by these emerging pathogens in a Saudi tertiary care hospital. All CoNS-positive cultures from inpatients were identified using the standard methods during a 10-month period. Antimicrobial susceptibility testing was done using the reference broth microdilution method. A total of 208 isolates were identified; of these 75 (32.2%) were considered infection associated, and 133 (67.8%) were considered contamination. S. epidermidis accounted for 34.7% of bacteremia cases, followed by S. hominis (21.3%), S. haemolyticus (16%), and S. saprophyticus (12%). Central venous catheters (p ≤ 0.0001), prior antibiotic therapy (p ≤ 0.0001), the occurrence of more than one positive blood culture (p ≤ 0.0001), and intensive care unit (ICU) admission (p = 0.007) were all independently associated with CoNS bacteremia. Overall, all isolates were highly resistant to penicillin (94.7%), oxacillin (90.7%), and erythromycin (85.3%). The rates of susceptibility to vancomycin, daptomycin, and teicoplanin were 98.7%, 98.7%, and 93.3%, respectively. Our results further highlight that accurate identification and susceptibility testing of CoNS isolates from nosocomial BSIs are crucial to minimize excessive antibiotic use and unnecessary catheter removal. In addition, daptomycin may be an efficient alternative therapeutic option for CoNS resistant to oxacillin and other commonly used antibiotics.
Artini, Marco; Cellini, Andrea; Papa, Rosanna; Tilotta, Marco; Scoarughi, Gian Luca; Gazzola, Simona; Fontana, Cecilia; Tempera, Gianna; Cocconcelli, Pier Sandro; Selan, Laura
Coagulase-negative staphylococci (CoNS) belong to saprophytic microbiota on the skin and mucous membranes of warm-blooded animals and humans, but are also isolated from foodstuffs such as meat, cheese, and milk. In other circumstances, some CoNS can act as pathogens. Thus the presence of CoNS may not be an immediate danger to public health, but can become a risk factor. In particular antibiotic-resistant genes could be transferred to other potentially pathogenic microorganisms. Furthermore, CoNS are known to be strong biofilm producers and this is also a risk factor for public health. The aim of the present work was to determine the genotypic and phenotypic profiles of 106 CoNS belonging to four different species isolated from five different Italian cheeses for the presence of some adhesion and virulence features. In order to verify a possible correlation between the formation of biofilm and staphylococcal virulence factors, we checked the presence of adhesin genes by PCR and we investigated the ability of these strains to make biofilm at different temperatures. Furthermore, in some conditions, we analyzed surface proteins and autolytic pattern of selected strains. In conclusion, we checked the presence of norA and mecA genes responsible for fluoroquinolones and methicillin resistance, respectively. We found resistant genes in a proportion of the food isolates in amounts of 9.4% (mecA) and 5.7% (norA). These data support the importance to continuously examine the microbiota not only for the creation of a database but also to safeguard public health. © The Author(s) 2015.
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
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
Sampimon, O.C.; Lam, T.G.J.M.; Mevius, D.J.; Schukken, Y.H.; Zadoks, R.N.
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,
van der Mei, HC; van de Belt-Gritter, B; Reid, G; Bialkowska-Hobrzanska, H; Busscher, HJ
Physico-chemical cell surface properties of 23 coagulase-negative staphylococcal strains, including contact angles, zeta potentials and elemental cell surface composition were measured, together with the adhesion of all strains to hexadecane, The data were employed in a hierarchical cluster
Nielsen, J; Kolmos, H J; Rosdahl, V T
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...... specimens were obtained from the skin at the insertion site and the hub, and blood cultures were obtained from the catheter. Among coagulase-negative staphylococci, S. epidermidis was the most frequently (80%) isolated species, and two biotypes accounted for 55.7% of the 41 biotypes isolated. 11 septicaemia...... cases due to coagulase-negative staphylococci occurred, all caused by S. epidermidis, and the incidence of S. epidermidis septicaemia was 21% among patients and 16% among catheter periods. S. epidermidis septicaemia occurred in 17%, 31% and 33% of all catheter periods in which S. epidermidis...
John, Joseph F; Harvin, Alexander M
Coagulase-negative staphylococci (CNS) are a heterogenous group of Gram-positive cocci that are widespread commensals among mammalia. Unlike their coagulase-positive counterpart, Staphylococcus aureus, CNS produce few virulence patterns and normally refrain from invading tissue. Yet, not only can CNS cause infections in normal host tissue, but modern medicine has also seen their rise as opportunists that display adherence to medical device materials to produce a protective biofilm. CNS have historically been more resistant to antimicrobials, including the beta-lactam antibiotics, than S. aureus and some hospitals reveal rates of oxacillin resistance in CNS approaching 90%. Cross resistance to non-beta-lactam agents has been a recurrent theme over the past 40 years in the CNS. Thus, there has been a pressing need for newer antimicrobial agents with good antistaphylococcal activity. Those new agents tend to have excellent antistaphylococcal activity include daptomycin, linezolid, oritavancin, telavancin, tigecycline, dalbavancin, new quinolones, and ceftibiprole, several of which have unique mechanisms of action. The MIC₉₀ for these new compounds typically ranges from 0.5-4 mug/mL. Staphylococcal biofilm formation is quite common in CNS infections and markedly increases the MIC for most older antimicrobials. Several of the newer agents offer some promise of penetration of biofilm to inhibit or kill adherent staphylococci. CNS will likely remain a major cause of infections in the modern age, evolve further antimicrobial resistance mechanisms, and require development of newer antimicrobials for curative therapy.
Holtzman, Carol; Whitney, Dana; Barlam, Tamar; Miller, Nancy S
Peptide nucleic acid fluorescence in situ hybridization (PNA FISH) was instituted at Boston Medical Center for the rapid identification of coagulase-negative staphylococci (CoNS). Without active notification or antimicrobial stewardship intervention, a pre- and postimpact analysis showed no benefit of this assay with respect to the length of hospital stay or vancomycin use.
Brokke, P.; Brokke, P.; Dankert, J.; Hogt, A.H.; Feijen, Jan
The cell surface hydrophobicity of 14 encapsulated and 21 non-encapsulated coagulase-negative staphylococci (CN staph) as determined with the salt aggregation test (SAT) as well as with the xylene-water method ranged widely. Non-encapsulated strains adhered well onto fluorinated
Performance of API Staph ID 32, Staph-Zym for identification of coagulase-negative staphylococci isolated from bovine milk samples NETHERLANDS (Sampimon, O.C.) NETHERLANDS Received: 2008-06-06 Revised: 2008-11-03 Accepted: 2008-11-06
Sousa, Margarida; Silva, Nuno; Igrejas, Gilberto; Sargo, Roberto; Benito, Daniel; Gómez, Paula; Lozano, Carmen; Manageiro, Vera; Torres, Carmen; Caniça, Manuela; Poeta, Patrícia
Wild animal populations in contact with antimicrobials and antimicrobial resistant bacteria that are daily released into the environment are able to become unintentional hosts of these resistant microorganisms. To clarify this issue, our study evaluated the presence of antibiotic resistance determinants on coagulase-negative staphylococci recovered from birds of prey and studied their genetic relatedness by pulsed-field gel electrophoresis (PFGE). The unusual vga(A) and erm(T) genes, which confer resistance to clindamycin and erythromycin, respectively, were detected in Staphylococcus sciuri or Staphylococcus xylosus strains and the tet(K) gene in Staphylococcus kloosii. The PFGE patterns showed that three S. xylosus (isolated of Strix aluco and Otus scops) and two S. sciuri (recovered from Strix aluco and Milvus migrans) were clonally indistinguishable. These animals could be a source of unusual antimicrobial resistance determinants for highly used antibiotics in veterinary clinical practice.
Mediano, Pilar; Fernández, Leonides; Jiménez, Esther; Arroyo, Rebeca; Espinosa-Martos, Irene; Rodríguez, Juan M; Marín, María
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).
Voronina, O L; Kunda, M S; Dmitrienko, O A; Liubasovskaia, L A; Kovalishena, O V; Popov, D A; Lunin, V G
Comparative analysis of species diversity of sample of coagulase-negative staphylococci (CNS) isolated in hospitals of different specializations. For identification of 102 CNS strains, biochemical systems manufactured by NPO "Diagnostic Systems", VITEK 2 Compact, and BBL Crystal as well as sequencing of fragments of tuf and gap genes were used. Greater differentiating capability of genotyping compared with phenotyping methods for species identification of staphylococci was demonstrated. Six CNS species were identified in the sample: S. epidermidis, S. haemolyticus, S. hominis, S. warneri, S. capitis, and S. pasteuri. The largest species diversity was noted for strains from maternity hospitals in Nizhny Novgorod and Kulakov Scientific Center for Obstetrics, Gynecology and Perinatology. Strains isolated from blood of patients in Bakulev Center for Cardiovascular Surgery were represented mostly by S. epidermidis and S. haemolyticus. Differences in species diversity of CNS--causative agents of neonatal conjunctivitis and omphalitis--were observed. Two species of CNS: S. epidermidis and S. haemolyticus pose special threat as nosocomial pathogens both in hospitals for adults and obstetrical facilities. Additionally, in neonatal units it is necessary to control such species as S. warneri, S. capitis, S. pasteuri.
Bexiga, Ricardo; Rato, Márcia G; Lemsaddek, Abdelhak; Semedo-Lemsaddek, Teresa; Carneiro, Carla; Pereira, Helena; Mellor, Dominic J; Ellis, Kathryn A; Vilela, Cristina L
The objectives of this study were to compare the impact of different coagulase-negative species (CNS) on udder health measured in terms of individual quarter milk somatic cell count (SCC) and duration of intramammary infection, and to get some insight into most likely routes of infection for different CNS species. This longitudinal observational study was performed on four farms that were sampled at 4-week intervals for a total of 12 visits each. Quarters infected with CNS were followed through time with milk samples being submitted for bacteriological culture and SCC determination. PCR amplification of the internal transcribed spacer region and sequencing of the sodA and rpoB genes were used for species allocation. Pulsed-field gel electrophoresis (PFGE) was performed to assess strain identity. The percentage of quarters affected per farm varied between 6 and 35%, with the most frequently isolated CNS species being Staphylococcus epidermidis, followed by Staph. simulans, Staph. chromogenes and Staph. haemolyticus. It was possible to follow 111 intramammary infections due to CNS through time. Duration of infection had a mean of 188 d and was not significantly different between CNS species. Geometric mean quarter SCC overall was 132 000 cells/ml and was also not significantly different between CNS species. Despite the possibility of a different epidemiology of infection, the impact in terms of udder health seems to be similar for different CNS species.
Aral, Murat; Keleş, Erol; Okur, Erdoğan; Alpay, H Cengiz; Yilmaz, Mustafa
To investigate the pathogenicity and antibiotic resistance of coagulase-negative staphylococci (CNS) isolated from the maxillary and ethmoid sinuses of patients undergoing endoscopic sinus surgery for chronic sinusitis. Ninety-three patients (63 males, 30 females) aged between 19 - 68 years, who had undergone functional endoscopic sinus surgery (FESS) for chronic sinusitis, were included in the study. Nasal mucosa, skin and adjacent structures were cleansed with povidone-iodine solution before surgery to prevent a probable contamination. In all patients, nasal swabs were taken before and after the application of povidone-iodine solution. Colonies isolated and identified as Staphylococci in cultures were further investigated for pathogenicity and antibiotic susceptibility. Slime test was used to determine the pathogenicity of CNS. The relationship between antibiotic resistance of pathogenic and non-pathogenic CNS was compared by chi2 analysis. While bacterial growth rate was 62.3% in nasal swab cultures taken before the application of povidone-iodine solution, it decreased to 12.9% after the application of solution. Microorganisms were isolated in 95.6% of cultures taken from maxillary sinuses and in 91.3% of cultures obtained from ethmoid sinuses during the FESS. The most frequently isolated microorganism in each of the sinuses was CNS. Slime test was carried out in 30 CNS isolated. Twelve of these were slime positive and 18 were slime negative. While 83.3% of CNS isolated was resistant to penicilin, all of CNS were sensitive to vancomycin and teikoplanine. The difference between slime positive and slime negative CNS for gentamicin and ciprofloxacin resistance was statistically significant (p<0.05). We consider that the pathogenicity tests like slime production and antimicrobial susceptibilities of CNS frequently isolated from the patients with chronic sinusitis should be investigated and also these microorganisms should be kept in mind in the selection of empiric
J. Sedef Göçmen
Full Text Available Objectives: In our study we investigated the effects ofparenteral nutrition (PN solutions on Coagulase negativestaphylococci (CoNS biofilm production.Materials and methods: Thirty nine CoNS strains isolatedfrom hemocultures and a reference strain (ATCC 12228Staphylococcus epidermidis were included. Bacterial dilutionswere made in Tryptic Soy Broth (TSB. The experimentalmediums were 1. Glucose, 2. Amino acid, 3. Lipid,4. Glucose+ Amino acid+ lipid, 5. Glucose+ Amino acid, 6.Glucose+ Lipid, 7. Amino acid+ Lipid, and 8. Control (TSB.Biofilm formation was evaluated by “quantitative microdilutionplaque test”. The values greater than cut off valueare considered as positive. Biofilm positivity was dividedinto 3 groups (mild, moderate and intensive and all otherstrains under cutoff value were accepted as negative. Thenumbers of biofilm positive strains derived from 1-7. mediumswere compared with each other, and with the resultsof control.Results: The three-component PN solution and two componentPN solutions containing glucose+ lipid and aminoacid+ lipid were found to increase the biofilm productionactivity of CoNS when compared to the control group.Slime positivity in medium 1 and 2 was lower than controlsignificantly, in medium 4, 6, and 7 slime positivity washigher considerably. The indifferent results were obtainedwithin the mediums 1, 2, 3 and within the mediums 4, 5, 6,and 7.Conclusions: In our study, it was found that, glucose, aminoacid and lipid solutions which were building structuresof PN decreased the biofilm production when used solitary.However use of the compounds increased the biofilmproduction. Therefore, we can conclude that PN solutionsgiven as mixtures in routine practice increase the risk ofcatheter infection. J Clin Exp Invest 2012; 3(4: 505-509Key words: Catheter-related infections, biofilm, parenteralnutrition
Dufour, S; Dohoo, I R; Barkema, H W; Descôteaux, L; Devries, T J; Reyher, K K; Roy, J-P; Scholl, D T
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
Soeorg, Hiie; Metsvaht, Tuuli; Eelmäe, Imbi; Metsvaht, Hanna Kadri; Treumuth, Sirli; Merila, Mirjam; Ilmoja, Mari-Liis; Lutsar, Irja
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.
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.
B. K. Bansal
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.
Smith, Peter A; Powers, Michael E; Roberts, Tucker C; Romesberg, Floyd E
The arylomycins are a class of natural-product antibiotics that act via the inhibition of type I signal peptidase (SPase), and we have found in diverse bacteria that their activity is limited by the presence of a resistance-conferring Pro residue in SPase that reduces inhibitor binding. We have also demonstrated that Staphylococcus epidermidis, which lacks this Pro residue, is extremely susceptible to the arylomycins. Here, to further explore the potential utility of the arylomycins, we report an analysis of the activity of a synthetic arylomycin derivative, arylomycin C₁₆, against clinical isolates of S. epidermidis and other coagulase-negative staphylococci (CoNS) from distinct geographical locations. Against many important species of CoNS, including S. epidermidis, S. haemolyticus, S. lugdunensis, and S. hominis, we find that arylomycin C₁₆ exhibits activity equal to or greater than that of vancomycin, the antibiotic most commonly used to treat CoNS infections. While the susceptibility was generally correlated with the absence of the previously identified Pro residue, several cases were identified where additional factors also appear to contribute.
Oliveira, Fernando; Lima, Cláudia Afonso; Brás, Susana; França, Ângela; Cerca, Nuno
Coagulase-negative staphylococci (CoNS) are common bacterial colonizers of the human skin. They are often involved in nosocomial infections due to biofilm formation in indwelling medical devices. While biofilm formation has been extensively studied in Staphylococcus epidermidis, little is known regarding other CoNS species. Here, biofilms from six different CoNS species were characterized in terms of biofilm composition and architecture. Interestingly, the ability to form a thick biofilm was not associated with any particular species, and high variability on biofilm accumulation was found within the same species. Cell viability assays also revealed different proportions of live and dead cells within biofilms formed by different species, although this parameter was particularly similar at the intraspecies level. On the other hand, biofilm disruption assays demonstrated important inter- and intraspecies differences regarding extracellular matrix composition. Lastly, confocal laser scanning microscopy experiments confirmed this variability, highlighting important differences and common features of CoNS biofilms. We hypothesized that the biofilm formation heterogeneity observed was rather associated with biofilm matrix composition than with cells themselves. Additionally, our results indicate that polysaccharides, DNA and proteins are fundamental pieces in the process of CoNS biofilm formation. © FEMS 2015. All rights reserved.
Bansal, B. K.; Gupta, D. K.; Shafi, T. A.; Sharma, S.
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. PMID:27047108
Bansal, B K; Gupta, D K; Shafi, T A; Sharma, S
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.
Semedo-Lemsaddek, Teresa; Carvalho, Laura; Tempera, Carolina; Fernandes, Maria H; Fernandes, Maria J; Elias, Miguel; Barreto, António S; Fraqueza, Maria J
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®
Ruaro, Angela; Andrighetto, Christian; Torriani, Sandra; Lombardi, Angiolella
The aim of the current study was to detect coagulase-negative staphylococci (CNS) in raw milk and cheeses produced in North Italy, and to analyze isolates for their biodiversity, safety aspects and technological properties. Molecular identification methods revealed a high biodiversity among isolates and assigned them to 17 species. The most recovered species were Staphylococcus equorum (12%), Staphylococcus lentus (12%), Staphylococcus simulans (12%), Staphylococcus sciuri (10%), and Staphylococcus xylosus (9%). The presence of ten transferable antibiotic resistance (AR) genes was verified by PCR and 19% of isolates were positive, with tet(K) being the most frequent gene (10%); interestingly, no strain carried multiple AR genes. Twenty-four isolates displayed hemolytic activity; tyrosine decarboxylase gene (tdcA) was found in two isolates, while histidine decarboxylase gene (hdcA) and enterotoxin genes (se) were not detected. Isolates were further characterized for the presence of some relevant technological properties; 16% of isolates displayed proteolytic activity and 39% lipolytic activity, while no one of the isolates was found to exhibit antimicrobial activity against Staphylococcus aureus. This study provided evidence of a low occurrence of safety hazards in CNS isolated from dairy products. Copyright © 2012 Elsevier Ltd. All rights reserved.
Fajardo Olivares, Miguel; Hidalgo Orozco, Rocío; Rodríguez Garrido, Saray; Gaona Álvarez, Cristina; Sánchez Silos, Rosa María; Hernández Rastrollo, Ramón; Martínez Tallo, Emilia; Cordero Carrasco, Juan Luis
Coagulase-negative-Staphylococci (CNS) are the major cause of bacteraemia and sepsis in newborns. CNS methicillin resistance and its loss of sensitivity to glycopeptide antibiotics, make treatment significantly more difficult in positive cocci infections. To study MIC vancomycin, teicoplanin and linezolid in different species of CNS methicillin resistant isolates from blood cultures from paediatric patients. Clinically relevant CNS methicillin resistant isolates from paediatric blood cultures from different hospitalization wards were tested. The isolates were identified by biochemical tests by means in the Combo panels 31 of MicroScan (Dade Behring, Siemens). Resistance to oxacillin and susceptibility to vancomycin, teicoplanin and linezolid were tested by microdilution panels as cited above. We also tested teicoplanin and linezolid sensitivity using Etest. 50 methicillin resistant strains were isolated: 37 (74%)S. epidermidis, 7 (14%) S. hominis, 4 (8%) S. haemolyticus and 2 (4%) Staphylococcus spp. 26 strains were observed with reduced susceptibility to vancomycin MIC = 2 mg/L, (22 S. epidermidis, 2 S. haemolyticus and 2 Staphylococcus spp.) and 21 strains with loss of susceptibility to teicoplanin, MIC = 4-16 mg/L (20 S. epidermidis and 1 S. haemolyticus). No CNS linezolid resistant was found. There is a linear correlation between increased vancomycin MIC and teicoplanin MIC. There is a statistically significant difference (p teicoplanin in the vancomycin group = 2 mg/L with respect to the vancomycin group ≤ 1 mg/L. We also observed very low levels of linezolid MIC for all strains.
Jeong, Do-Won; Lee, Bitnara; Her, Jae-Young; Lee, Kwang-Geun; Lee, Jong-Hoon
To select starters for the production of meju and doenjang, traditional Korean fermented soybean foods, we assessed the safety and technological properties of their predominant isolates, Staphylococcus saprophyticus, Staphylococcus succinus and Staphylococcus xylosus. Phenotypic antibiotic resistance, hemolysis and biofilm formation were strain-specific. None of the S. succinus isolates exhibited antibiotic resistance or hemolytic activities. Thirty-three selected strains, identified through safety assessments of 81 coagulase-negative staphylococci (CNS) isolates, produced cadaverine, putrescine, and tyramine, but not histamine in the laboratory setting. The production of these three biogenic amines may, however, be insignificant considering the high levels of tyramine produced by the control, Enterococcus faecalis. The 33 CNS strains could grow on tryptic soy agar containing 21% NaCl (w/v), exhibited acid producing activity at 15% NaCl, and expressed strain-specific protease and lipase activities. S. succinus 14BME1, the selected starter candidate, produced significant amounts of benzeneacetic acid, 2,3-butanediol, trimethylpyrazine, and tetramethylpyrazine through soybean fermentation. Copyright © 2016 Elsevier B.V. All rights reserved.
Janssens, M; Van der Mijnsbrugge, A; Sánchez Mainar, M; Balzarini, T; De Vuyst, L; Leroy, F
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.
Bexiga, Ricardo; Ellis, Kathryn A; Vilela, Cristina L; Mellor, Dominic J
Coagulase-negative staphylococci (CNS) are the most frequently isolated bacteria from milk samples in several studies worldwide. Despite their relative frequency, specific measures aiming at their control are not well established. One possible measure to include in a control programme is lactational antimicrobial treatment. The decision to perform such treatment, as well as other actions on farm, should be based on the likelihood of financial return. A deterministic model was used to evaluate whether performing an antimicrobial treatment during the lactation for quarters infected with CNS was financially justifiable. Input variables for the impact of CNS on udder health were based on a previous study by the same authors and on available literature on the subject. Prices included in the model were based on 2009/2010 conditions in Portugal. The average result per antimicrobial treated quarter was a net loss of €38·74. Performing a sensitivity analysis to evaluate how systematic variation of the input variables of the model would lead to outcome changes showed that variation in input variables nearly always led to a negative outcome, with the greatest variation in losses observed for variation in the length of treatment and milk withdrawal period (-€46·26 to -€28·49). The situations in which a net benefit was to be expected included the bulk tank somatic cell count decreasing to a level corresponding to a premium payment or to penalties being avoided, and the prevention of transmission of CNS in the milking parlour when the possibility of transmission was at its highest level. For most situations, lactational treatment of CNS subclinical mastitis was not financially justifiable.
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
Dolder, C; van den Borne, B H P; Traversari, J; Thomann, A; Perreten, V; Bodmer, M
Bacteriological status, evaluation of udder symmetry, udder hygiene, and teat end scores of 92 dairy cows were assessed on 3 Swiss dairy farms in a longitudinal 1-yr study to determine risk factors for intramammary infection (IMI) with coagulase-negative staphylococci (CNS) species. Farm visits were performed monthly including sterile quarter milk sampling and udder evaluation of all lactating cows. Milk samples were evaluated for the presence of staphylococci using selective agar plates. Species identification was performed using MALDI-TOF mass spectrometry. Intramammary infection was defined as milk samples having ≥100 cfu per mL of milk according to culture results. Overall, 3,151 quarter samples were included in the statistical analysis. Staphylococcus chromogenes, Staphylococcus haemolyticus, Staphylococcus xylosus, and a Staphylococcus warneri-like species were the 4 most prevalent CNS species found. Hierarchical multivariable logistic regression models were built to evaluate risk factors for species-specific CNS IMI. Risk factors for Staph. chromogenes IMI were presence in herd B, the period from June 2014 to August 2014 and December 2014 to February 2015, and presence of udder edema. For Staph. haemolyticus, the relevant risk factor included coinfection with Staph. xylosus coinfection with other than the above-mentioned CNS species ("others") and the period from June 2014 to November 2014. Coinfection with Staph. haemolyticus and "others," the periods from June 2014 to August 2014 and December 2014 to February 2015, early phase of lactation (1-60 d in milk), and belonging to herd B were significantly associated with Staph. xylosus IMI. Mid and late lactation, coinfection with Staph. xylosus, and the period September 2014 to May 2015 were identified as significant risk factors for Staph. warneri-like IMI. For Staph. chromogenes, 60.6 and 26% of the variance was observed at the quarter and cow level, respectively, whereas for the other investigated species
Full Text Available Methicillin-resistant coagulase negative staphylococci (MR-CoNS are the major cause of infectious diseases because of their potential ability to form biofilm and colonize the community or hospital environments. This study was designed to investigate the biofilm producing ability, and the presence of mecA, icaAD, bap and fnbA genes in MR-CoNS isolates. The MR-CoNS used in this study were isolated from various samples of community environment and five wards of hospital environments, using mannitol salt agar (MSA supplemented with 4 μg/ml of oxacillin. The specie level of Staphylococcus haemolyticus, Staphylococcus epidermidis, Staphylococcus hominis and Staphylococcus warneri was identified by specific primers of groESL (S. haemolyticus, rdr (S. epidermidis and nuc (S. hominis and S. warneri. The remainder isolates were identified by tuf gene sequencing. Biofilm production was determined using Congo red agar (CRA and Microtiter plate (MTP assay. The mecA and biofilm associated genes (icaAD, fnbA and bap were detected using PCR method. From the 558 samples from community and hospital environments, 292 MR-CoNS were isolated (41 from community environments, and 251 from hospital environments. S. haemolyticus (41.1% and S. epidermidis (30.1% were the predominant species in this study. Biofilm production was detected in 265 (90.7% isolates by CRA, and 260 (88.6% isolates were detected by MTP assay. The staphylococci isolates derived from hospital environments were more associated with biofilm production than the community-derived isolates. Overall, the icaAD and bap genes were detected in 74 (29.5% and 14 (5.6% of all isolates from hospital environments. When tested by MTP, the icaAD gene from hospital environment isolates was associated with biofilm biomass. No association was found between bap gene and biofilm formation. The MR-CoNS isolates obtained from community environments did not harbor the icaAD and bap genes. Conversely, fnbA gene presented
Seng, Rathanin; Kitti, Thawatchai; Thummeepak, Rapee; Kongthai, Phattaraporn; Leungtongkam, Udomluk; Wannalerdsakun, Surat; Sitthisak, Sutthirat
Methicillin-resistant coagulase negative staphylococci (MR-CoNS) are the major cause of infectious diseases because of their potential ability to form biofilm and colonize the community or hospital environments. This study was designed to investigate the biofilm producing ability, and the presence of mecA, icaAD, bap and fnbA genes in MR-CoNS isolates. The MR-CoNS used in this study were isolated from various samples of community environment and five wards of hospital environments, using mannitol salt agar (MSA) supplemented with 4 μg/ml of oxacillin. The specie level of Staphylococcus haemolyticus, Staphylococcus epidermidis, Staphylococcus hominis and Staphylococcus warneri was identified by specific primers of groESL (S. haemolyticus), rdr (S. epidermidis) and nuc (S. hominis and S. warneri). The remainder isolates were identified by tuf gene sequencing. Biofilm production was determined using Congo red agar (CRA) and Microtiter plate (MTP) assay. The mecA and biofilm associated genes (icaAD, fnbA and bap) were detected using PCR method. From the 558 samples from community and hospital environments, 292 MR-CoNS were isolated (41 from community environments, and 251 from hospital environments). S. haemolyticus (41.1%) and S. epidermidis (30.1%) were the predominant species in this study. Biofilm production was detected in 265 (90.7%) isolates by CRA, and 260 (88.6%) isolates were detected by MTP assay. The staphylococci isolates derived from hospital environments were more associated with biofilm production than the community-derived isolates. Overall, the icaAD and bap genes were detected in 74 (29.5%) and 14 (5.6%) of all isolates from hospital environments. When tested by MTP, the icaAD gene from hospital environment isolates was associated with biofilm biomass. No association was found between bap gene and biofilm formation. The MR-CoNS isolates obtained from community environments did not harbor the icaAD and bap genes. Conversely, fnbA gene presented in MR
Sánchez Mainar, María; Stavropoulou, Despoina Angeliki; Leroy, Frédéric
The production of fermented meats, such as fermented sausage, relies on the metabolic activities of lactic acid bacteria and catalase-positive cocci, in particular the group of coagulase-negative staphylococci (CNS). Conventional use of CNS as meat starter cultures usually leads to an appropriate cured colour development based on their nitrate reductase activity, whereas their catalase activity reduces oxidative damage. In addition, CNS metabolism contributes to flavour, although the precise effects are difficult to estimate. There are reasons to believe that these basic technological features of CNS can be further enlarged by exploring their full metabolic potential. Non-negligible differences in metabolism among and within different species of CNS indicate that a rational selection of strains may lead to the development of novel starter cultures with enhanced functionality. Firstly, the use of CNS strains with a superior ability to use alternative energy sources, such as nucleosides or arginine, may improve culture competitiveness and survival. Secondly, cured colour generation could be optimised to lower the amounts of curing salts needed, either by selecting for efficient nitrate-reducing CNS strains or by exploring the potential alternative based on nitric oxide synthase activity. Thirdly, CNS with specific aroma-producing abilities may help to accentuate specific flavours, whereby the selection of wild-type strains from artisan-type fermented sausages seems attractive in the framework of innovation-through-tradition. Finally, bacteriocin-producing CNS strains may offer solutions for bioprotection towards meat pathogens such as Clostridium botulinum and Staphylococcus aureus. Overall, making use of the metabolic inter- and intraspecies heterogeneity of CNS is promising for the elaboration of healthier, tastier, and safer fermented meats. Yet, the proposed strategies are sometimes still overly theoretical and speculative, requiring further proof
Garza-González, Elvira; López, Daniel; Pezina, Cesar; Muruet, Walter; Bocanegra-García, Virgilio; Muñoz, Ivan; Ramírez, Camilo; Llaca-Díaz, Jorge M
The objective of this study was to determine the distribution of staphylococcal cassette chromosome mec (SCCmec) elements in meticillin-resistant coagulase-negative staphylococci (MR-CoNS) isolated from a tertiary-care hospital in Mexico and to examine the relationship to drug resistance. Fifty selected MR-CoNS isolates collected from catheters (n=15), blood (n=15), bone (n=9), bronchial lavage (n=2) and urine (n=2) and one isolate each from an abscess, cerebrospinal fluid, eye, pleural effusion, synovial fluid, tracheal aspirate and wound secretion were examined. Susceptibility testing was performed by the broth microdilution method. SCCmec types were determined by multiplex PCR and PFGE was carried out as described previously for Staphylococcus aureus. Among the MR-CoNS strains studied, the most frequently isolated species were Staphylococcus epidermidis (n=26) and Staphylococcus haemolyticus (n=13). Staphylococcus cohnii (n=5), Staphylococcus hominis (n=3), Staphylococcus sciuri (n=1), Staphylococcus pasteuri (n=1) and the recently described species Staphylococcus pettenkoferi (n=1) were also identified. The most frequent MR-CoNS genotype identified was SCCmec type IVa in S. epidermidis isolates, which also showed a high diversity in their PFGE patterns. A clone was found that amplified both SCCmec III and V elements in five isolates examined. The single MR S. pettenkoferi isolate harboured SCCmec type IVd and the single MR S. pasteuri isolate harboured SCCmec type I. The carriage of SCCmec type III was associated with resistance or intermediate resistance to meropenem (P IVa and the high genetic diversity among MR-CoNS strains. As far as is known, this is the first report describing the newly identified S. pettenkoferi possessing SCCmec IVd and S. pasteuri harbouring SCCmec type I. MR-CoNS harbouring SCCmec type III were found to be more resistant to meropenem.
Breyne, K; De Vliegher, S; De Visscher, A; Piepers, S; Meyer, E
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
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
Adilson De Oliveira
Full Text Available Background Coagulase-negative staphylococci (CoNS are one of the most frequent causative agents of neonatal nosocomial infections, especially in premature and low-weight newborns. Risk factors for infection include extracellular polysaccharide production and consequent biofilm formation that permit adhesion to the smooth surface of catheters and other medical devices. The objective of this study was to identify CoNS strains isolated from 105 newborns admitted to the Neonatal Unit of our hospital, and to evaluate the association of biofilm production and host risk factors with the occurrence of infection. Methods CoNS isolates were identified and classified as significant or contaminant based on clinical and laboratory data of the newborn medical records. Perinatal risk factors for infection, neonatal clinical evolution, and antibiotic treatment were analysed. In addition, the presence of genes ( icaA, icaC and icaD responsible for biofilm production in CoNS was investigated. Results Among the 130 CoNS strains studied, 66 (50.8% were classified as clinically significant and 64 (49.2% as contaminant. There was no difference in the detection of biofilm-specific genes between CoNS strains isolated from newborns with (81.8% and without infection (84.3%, although 11 (91.7% of the 12 children whose death was related to CoNS were infected with strains that were positive for these genes. Forty-five (83.3% of the 54 newborns infected with CoNS were premature and 33 (61.1% had a birth weight ≤ 1,500 g. Most newborns infected with CoNS had been submitted to invasive procedures, including catheter use (85.2%, parenteral nutrition (61.1%, and mechanical ventilation (57.4%. S. epidermidis was the most frequently isolated species (81.5% and was more related to infection (86.3% than to contamination (76.5%. Conclusion Most newborns infected with CoNS presented factors that contributed to the colonization and development of infection with these microorganisms
Development and evaluation of hexaplex PCR for rapid detection of methicillin, cadmium/zinc and antiseptic-resistant staphylococci, with simultaneous identification of PVL-positive and -negative Staphylococcus aureus and coagulase negative staphylococci.
Panda, Sasmita; Kar, Sarita; Choudhury, Ranginee; Sharma, Savitri; Singh, Durg V
We developed a multiplex PCR to detect the presence of methicillin- (mecA), cadmium/zinc-(czrC) and antiseptic-resistant (qacA/B) staphylococci and to identify Panton-Valentine leukocidin (PVL)-positive and -negative Staphylococcus aureus and coagulase-negative staphylococci (CoNS) from infected and healthy eyes. The assay was validated on 177 staphylococci comprising of 55 each of S. aureus and CoNS isolated from infected eyes and five S. aureus and 62 CoNS isolated from healthy eyes and nine direct ocular samples. Nine direct ocular samples for in situ testing consisted of corneal scrapings (4), conjunctiva swabs (2) and others (3). Multiplex PCR result was correlated with genotype data obtained with single PCR and dot-blot assay. The control strains that were positive in multiplex PCR for 16S rRNA, nuc, mecA, pvl, czrC and qacA/B genes were also positive in the dot-blot assay. The specificity of amplified genes obtained with reference strains was further confirmed by DNA sequencing. The single step-hexaplex PCR method can be used for rapid detection of mecA, nuc, pvl, czrC and qacA/B genes in staphylococci with simultaneous identification of PVL-positive and -negative S. aureus and CoNS from a variety of ocular samples. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.
Mcdivitt, Maxine E.; Jerome, Norge W.
A fibrinogen-polymyxin medium and Staphylococcus Medium 110 were used in the isolation of coagulase-positive staphylococci in raw milk. Results indicated that both media allow the growth of some rods and of many coagulase-negative cocci. A significantly greater number of coagulase-positive staphylococci were identified by the tube test than were revealed by halo formation on fibrinogen-polymyxin medium. PMID:14325871
Abimanyu, Nagarajan; Krishnan, Arunkumar; Murugesan, Saravanan; Subramanian G, Kaushik; Gurumurthy, Sivakumar; Krishnan, Padma
Introduction: Methicillin-Resistant Staphylococcus aureus (MRSA) has become a major public health problem in both hospitals and communities. Panton – Valentine Leucocidin (PVL) has been reported to be an important marker for the highly pathogenic community acquired S. aureus infections. A rapid detection of these MRSA is very important for its treatment. The specific detection of MRSA is always a problem due to the prevalence of methicillin resistance among the coagulase negative Staphylococc...
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
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.
Chajęcka-Wierzchowska, Wioleta; Zadernowska, Anna; Nalepa, Beata; Sierpińska, Magda; Łaniewska-Trokenheim, Łucja
The aim of this work was to study the pheno- and genotypical antimicrobial resistance profile of coagulase negative staphylococci (CoNS) isolated from 146 ready-to-eat food of animal origin (cheeses, cured meats, sausages, smoked fishes). 58 strains were isolated, they were classified as Staphylococcus xylosus (n = 29), Staphylococcus epidermidis (n = 16); Staphylococcus lentus (n = 7); Staphylococcus saprophyticus (n = 4); Staphylococcus hyicus (n = 1) and Staphylococcus simulans (n = 1) by phenotypic and genotypic methods. Isolates were tested for resistance to erythromycin, clindamycin, gentamicin, cefoxitin, norfloxacin, ciprofloxacin, tetracycline, tigecycline, rifampicin, nitrofurantoin, linezolid, trimetoprim, sulphamethoxazole/trimethoprim, chloramphenicol, quinupristin/dalfopristin by the disk diffusion method. PCR was used for the detection of antibiotic resistance genes encoding: methicillin resistance--mecA; macrolide resistance--erm(A), erm(B), erm(C), mrs(A/B); efflux proteins tet(K) and tet(L) and ribosomal protection proteins tet(M). For all the tet(M)-positive isolates the presence of conjugative transposons of the Tn916-Tn1545 family was determined. Most of the isolates were resistant to cefoxitin (41.3%) followed by clindamycin (36.2%), tigecycline (24.1%), rifampicin (17.2%) and erythromycin (13.8%). 32.2% staphylococcal isolates were multidrug resistant (MDR). All methicillin resistant staphylococci harboured mecA gene. Isolates, phenotypic resistant to tetracycline, harboured at least one tetracycline resistance determinant on which tet(M) was most frequent. All of the isolates positive for tet(M) genes were positive for the Tn916-Tn1545 -like integrase family gene. In the erythromycin-resistant isolates, the macrolide resistance genes erm(C) or msr(A/B) were present. Although coagulase-negative staphylococci are not classical food poisoning bacteria, its presence in food could be of public health significance due to the possible spread of
Antimicrobial Resistance Pattern and Minimum Inhibitory Concentration of Vancomycin among Staphylococcus aureus and Coagulase-Negative Staphylococci Isolated from Clinical Specimens of Children in Tabriz
Shahram Abdoli Oskouie
Full Text Available Background & Objectives: Staphylococci are among common causes of community acquired and nosocomial infections around the world. Over the last decade, the resistance of these bacteria in hospital environments is increasing to various antibiotics such as vancomycin. The aim of present study was to determine the antimicrobial resistance pattern and Minimum Inhibitory Concentration (MIC values among a clinical collection of staphylococci isolated from hospitalized children in Tabriz. Methods: In this prospective and descriptive study, 88 staphylococcal isolates including 53 S. aureus and 35 coagulase-negative staphylococcus species were recovered from various clinical specimens referred to microbiology laboratory of Children Hospital during study period (April 2011 to March 2012. Susceptibility of the isolates against 15 different antimicrobial agents and MIC values of vancomycin was tested using standard disk diffusion and E-test methods respectively. Results: According to the results of drug susceptibility testing, vancomycin and rifampin were the most effective but clindamycin and penicillin were the least effective drugs against tested isolates. Accordingly, the prevalence of methicillin resistant Staphylococcus aureus (MRSA strains was determined more than 80%. According to MIC values, 13.2% of S. aureus and 3.3% of coagulase-negative staphylococcus isolates showed intermediate resistance to vancomycin. None of the isolates was fully resistant to vancomycin isolates in this study. Conclusion: Although fully vancomycin resistant staphylococci was not found among tested isolates in this study, there was VISA strains. Since there are reports on the emergence of VRSA strains from Iran and other countries, it is necessary for the clinician to care in prescription of vancomycin as a selective drug against staphylococcal infections. Moreover, the necessity of MIC measurement in determining of vancomycin susceptibility is more apparent.
De Visscher, A; Haesebrouck, F; Piepers, S; Vanderhaeghen, W; Supré, K; Leroy, F; Van Coillie, E; De Vliegher, S
This study aimed at testing the applicability of mannitol salt agar (MSA), a medium generally used in human medicine for differentiating Staphylococcus aureus from coagulase-negative staphylococci (CNS), for culturing bovine-associated CNS species. All test isolates from a comprehensive collection of well-identified CNS species, including both reference strains and field isolates, were able to grow. Subsequently, bulk milk samples and teat apex swabs were used to examine the capability of MSA for yielding CNS under field conditions. Sixty-nine and 47 phenotypically different colonies were retrieved from bulk milk and teat apices, respectively. The majority of isolates from teat apices were staphylococci, whereas in bulk milk, staphylococci formed a minority. After 24h of growth, recovery of separate colonies of CNS was much more convenient on MSA compared to a non-selective blood agar. The results of this study indicate that MSA is a suitable medium for both growth and recovery of bovine-associated CNS. Copyright © 2013 Elsevier Ltd. All rights reserved.
Mahmmod, Yasser Saadeldien Ibrahim; Svennesen, Line; Pedersen, Karl
Coagulase-negative staphylococci (CNS) frequently colonize teat skin and are common findings cultured from milk samples of cows with subclinical intramammary infections (IMI). Several species are related to IMI, but knowledge about the epidemiology of CNS species is limited. Cows in automatic...... milking systems (AMS) may have increased risk for teat colonization and IMI because more than 60 cows are milked several times daily with the same milking unit. The objectives of this study are (1) to investigate patterns of CNS species in milk samples and teat skin swabs in nine AMS herds and (2......) to identify the predisposing cow level risk factors for specific CNS IMI and teat colonization.In each herd, 30- 40 cows with somatic cell counts > 200,000 cells/ml in the previous milk recording are randomly selected and teat skin swabs and aseptic quarter foremilk samples are taken. Teat skin swabs...
Osman, Kamelia; Alvarez-Ordóñez, Avelino; Ruiz, Lorena; Badr, Jihan; ElHofy, Fatma; Al-Maary, Khalid S; Moussa, Ihab M I; Hessain, Ashgan M; Orabi, Ahmed; Saad, Alaa; Elhadidy, Mohamed
The objectives of this study were to characterize the diversity and magnitude of antimicrobial resistance among Staphylococcus species recovered from imported beef meat sold in the Egyptian market and the potential mechanisms underlying the antimicrobial resistance phenotypes including harboring of resistance genes (mecA, cfr, gyrA, gyrB, and grlA) and biofilm formation. The resistance gene mecA was detected in 50% of methicillin-resistant non-Staphylococcus aureus isolates (4/8). Interestingly, our results showed that: (i) resistance genes mecA, gyrA, gyrB, grlA, and cfr were absent in Staphylococcus hominis and Staphylococcus hemolyticus isolates, although S. hominis was phenotypically resistant to methicillin (MR-non-S. aureus) while S. hemolyticus was resistant to vancomycin only; (ii) S. aureus isolates did not carry the mecA gene (100%) and were phenotypically characterized as methicillin- susceptible S. aureus (MSS); and (iii) the resistance gene mecA was present in one isolate (1/3) of Staphylococcus lugdunensis that was phenotypically characterized as methicillin-susceptible non-S. aureus (MSNSA). Our findings highlight the potential risk for consumers, in the absence of actionable risk management information systems, of imported foods and advice a strict implementation of international standards by different venues such as CODEX to avoid the increase in prevalence of coagulase positive and coagulase negative Staphylococcus isolates and their antibiotic resistance genes in imported beef meat at the Egyptian market.
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
Lin, Yu-Tzu; Hung, Wei-Chun; Tsai, Jui-Chang; Leong, Kin Hong; Chen, Hsiao-Jan; Hsueh, Po-Ren; Teng, Lee-Jene
We examined the fusidic acid resistance determinants fusB, fusC, fusD, and fusF in coagulase-negative staphylococci (CoNS) clinical isolates. Among 208 resistant isolates, the fusB gene was the most common determinant in each species except in S. hominis subsp. hominis or in species carrying fusD or fusF. In S. hominis subsp. hominis, the fusC gene was the major determinant responsible for fusidic acid resistance. To understand the genetic context of fusC in S. hominis subsp. hominis, 31 fusC-positive S. hominis subsp. hominis isolates were examined. Among those isolates, 14 carried SCCfusC, 3 carried an SCC 476 -like element, and 8 carried a new SCC structure, SCC 3390 . As shown by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) analyses, the S. hominis subsp. hominis clinical isolates showed limited clonality. Taken together, SCCfusC has been found in S. hominis subsp. hominis, S. epidermidis, S. haemolyticus, S. capitis subsp. ureolyticus and S. aureus, suggesting its wide distribution and spreading among different species of staphylococci. Copyright © 2018 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.
Casaes Nunes, Raquel Soares; Pires de Souza, Camilla; Pereira, Karen Signori; Del Aguila, Eduardo Mere; Flosi Paschoalin, Vânia Margaret
Minas Frescal is a typical Brazilian fresh cheese and one of the most popular dairy products in the country. This white soft, semiskimmed, nonripened cheese with high moisture content is obtained by enzymatic coagulation of cow milk using calf rennet or coagulants, usually in industrial dairy plants, but is also manufactured in small farms. Contamination of Minas Frescal by several staphylococci has been frequently reported. Coagulase-negative staphylococci (CNS) strains are maybe the most harmful, as they are able to produce heat-stable enterotoxins with super antigenic activities in food matrices, especially in dairy products such as soft cheeses. The aim of the present study was to investigate the presence of CNS strains in Minas Frescal marketed in southeastern Brazil concerning the risk of staphylococci food poisoning by the consumption of improperly manufactured cheese and the possibility of these food matrices being a reservoir of staphylococcal resistance to antimicrobials. Ten distinct CNS strains were found in 6 cheeses from distinct brands. The most frequent species were Staphylococcus saprophyticus (40%), Staphylococcus xylosus (30%), Staphylococcus sciuri (20%), and Staphylococcus piscifermentans (10%). Three strains were identified to the Staphylococcus genera. Three major species groups composed of 3 refined clusters were grouped by phylogenetic analyses with similarities over to 90%. All CNS strains carried multiple enterotoxin genes, with high incidence of sea and seb (90 and 70%, respectively), followed by sec/see, seh/sei, and sed with intermediate incidence (60, 50, and 40%, respectively), and, finally, seg/selk/selq/selr and selu with the lowest incidence (20 and 10%, respectively). Real-time reverse transcription PCR and ELISA assays confirmed the enteroxigenic character of the CNS strains, which expressed and produced the enterotoxins in vitro. The CNS strains showed multiresistance to antimicrobial agents such as β-lactams, vancomycin, and
Decousser, Jean-Winoc; Desroches, Marine; Bourgeois-Nicolaos, Nadège; Potier, Julien; Jehl, François; Lina, Gérard; Cattoir, Vincent; Vandenesh, François; Doucet-Populaire, Florence
Multiresistance in staphylococci constitutes a major challenge for the antimicrobial chemotherapy of invasive infections such as bacteraemia or bone and joint infections (BJIs). A nationwide prospective study was performed to detect antimicrobial resistance trends among staphylococci causing invasive infections. Between October 2011 and February 2012, 367 meticillin-resistant Staphylococcus aureus (MRSA) and 695 coagulase-negative staphylococci (CoNS) were collected from 37 French hospitals, mainly from bacteraemia (59.9%) and osteoarticular infections (29.0%). Minimum inhibitory concentrations (MICs) were determined by broth microdilution, and specific screening and confirmation tests were performed to detect heterogeneous vancomycin-intermediate S. aureus (hVISA). Staphylococcal isolates exhibiting a linezolid MIC>4 mg/L were further characterised to determinate their clonal relationships and the mechanism of resistance. MRSA exhibited additional resistances, including levofloxacin (82% associated resistance), gentamicin (13.6%), fusidic acid (13.6%) and rifampicin (6.5%), compromising oral step-down therapy in BJIs. Only two hVISA strains (0.5%) were identified. Among the CoNS, mainly Staphylococcus epidermidis (506/695; 72.8%), resistance to first- and second-line agents was more common. Linezolid resistance was identified in 10 CoNS (1.4%). The most frequent linezolid resistance mechanism was the G2576T mutation in 23S rDNA (9/10). For the first time in France, the cfr gene was found in five related sequence type 2 (ST2) S. epidermidis from two different hospitals, in association with ribosomal RNA and L3 ribosomal protein mutations. These national data must be considered when selecting empirical treatment for invasive staphylococcal infections. Moreover, the emergence and spread of linezolid-resistant CoNS carrying the cfr gene is of concern. Copyright © 2015 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.
Sieradzki, Krzysztof; Villari, Paolo; Tomasz, Alexander
Of 41 methicillin-resistant coagulase-negative staphylococcal clinical isolates collected during a 5-month period between late 1995 and early 1996, 28 showed tube dilution teicoplanin MICs of 4 to 8 μg/ml which increased to 16 to 32 μg/ml upon prolonged incubation. Cultures of such bacteria were heterogeneous; they contained subpopulations with frequencies of 10−5 to 10−4 that could grow on up to 50 μg of teicoplanin per ml. The same cultures were also heterogeneous with respect to susceptibility to vancomycin; while the MICs for the majority of cells were 2 to 4 μg/ml, subpopulations that could grow on 6 to 12 μg of vancomycin per ml were also present at frequencies of 10−5 to 10−7. Selective enrichment of such cultures for the resistant subpopulation occurred with relative ease under laboratory conditions. Heterogeneous phenotypes for teicoplanin (but not for vancomycin) susceptibility were also identified in several Staphylococcus epidermidis isolates collected during the preantibiotic era. The addition of half the MIC of teicoplanin inhibited autolysis and caused formation of cellular aggregates which disintegrated to individual bacteria in the stationary phase when the titer of teicoplanin in the medium fell to undetectable levels, indicating removal of the antibiotic from the culture medium by the bacteria. PMID:9449268
Renneberg, J; Rieneck, K; Gutschik, E
The purpose of the investigation was to evaluate two commercially available identification systems: a new modification of the Staph-Zym system (Rosco, Tåstrup, Denmark) and the Staph ID 32 API system (API System, BioMérieux, Paris, France). A local standard method to be used in routine laboratories was also evaluated. A total of 200 staphylococcal isolates, including strains from both the American Type Culture Collection and the Czechoslovak Collection of Microorganisms as well as 89 clinical isolates, were used in tests of all three identification systems. The Staph ID 32 API system identified from 50 to 100% of the reference strains and 82.1% of the clinical isolates correctly. The Staph-Zym system identified from 90 to 100% of the reference strains and 82.1% of the clinical isolates correctly. Most misidentifications were of minor importance, but in both systems major failures appeared (Staphylococcus aureus was identified as a coagulase-negative staphylococcus). Both systems needed backup from a reference laboratory to determine if two isolates were of the same strain. PMID:7615720
Hammad, Ahmed M; Watanabe, Wataru; Fujii, Tomoko; Shimamoto, Tadashi
Staphylococci are not part of the normal fish microflora. The presence of staphylococci on fish is an indication of (a) post-harvest contamination due to poor personnel hygiene, or (b) disease in fish. The aim of this study was to determine the prevalence, molecular genetic characteristics, antibiotic resistance and virulence factors of methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MR-CoNS) isolated from 200 samples of retail ready-to-eat raw fish (sashimi) collected from the Japanese prefecture of Hiroshima. We characterized 180 staphylococcal strains. A majority of the grocery stores surveyed (92%, 23/25) contained fish contaminated with Staphylococcus species. We recovered 175 S. aureus isolates from 174 (87%, 174/200) samples, with 170 isolates of MSSA. For the MRSA and MR-CoNS, 10 isolates were obtained from 10 samples (5%, 10/200) collected from 10 shops (40%, 10/25) belonging to four supermarket chains. SCCmec typing revealed the presence of a type IV.1 SCCmec cassette in S. warneri isolates, a type II.1 SCCmec cassette in S. haemolyticus isolates and a cassette in methicillin-resistant S. aureus (MRSA) isolates that could not be typed. Molecular typing of two MRSA isolates by spa sequencing and multilocus sequence typing (MLST) identified t1767 and ST8, respectively. Antibiotic resistance genes that confer resistance to aminoglycosides, tetracyclines, β-lactams, macrolides, lincosamides and streptogramin B (MLS(B)) antibiotics were detected. Genes encoding one or more of the following virulence factors: staphylococcal enterotoxins (seb, and sed), toxic shock syndrome toxin 1 (tst), exfoliative toxin (etaA) were detected in 14.2% (25/175) of S. aureus isolates. The accessory gene regulator (agr) typing of S. aureus isolates revealed that agr type 1 was most prevalent (96.5%, 169/175) followed by type 2 (2.2%, 4/175) and type 3 (1.1%, 2/175). None of
Bochniarz, M; Wawron, W
The aim of the present study was to evaluate the antibiotic susceptibility of methicillin-susceptible (MS) and methicillin-resistant (MR) coagulase-negative Staphylococcus (CNS) strains isolated from milk of cows with mastitis. The study was conducted on 100 CNS strains (20 MRCNS and 80 MSCNS) isolated from milk samples of 86 cows from the Lublin (Poland) region farms. Antibiotic susceptibility of microorganisms was evaluated using the disc-diffusion method on the Mueller-Hinton agar according to the guidelines of the National Committee for Clinical Laboratory Standards (NCCLS). The highest efficacy against MSCNS was demonstrated for cephalosporin antibiotics, i.e. cefacetril (91.3%), ceftiofur (67.5%), cefoperazone (66.3%) and cephalexin (60.0% of susceptible MSCNS strains). Moreover, a high percentage of vancomycin-susceptible strains was demonstrated (83.8%). The activity of combination of amoxicillin with clavulanic acid and gentamicin was found weaker (63.8% and 61.3% of susceptible strains, respectively). About 50.0% of MSCNS were susceptible to erythromycin, enrofloxacine and amoxicillin. A large proportion of CNS was resistant to neomycin, penicillin, tetracycline, streptomycin, lincomycin and ampicillin (28.8%, 30.0%, 31.3%, 31.3%, 33.8% and 33.8% of susceptible strains, respectively). The highest percentage of MRCNS was susceptible to vancomycin (75.0%), erythromycin (65.0%) and streptomycin (50.0%). Their susceptibility to enrofloxacine (35.0%) as well as gentamicin and tetracycline (30.0%) was markedly lower. The lowest activity was found for lincomycin and neomycin (20.0% of susceptible MRCNS strains, each).
Loonen, A J M; Jansz, A R; Kreeftenberg, H; Bruggeman, C A; Wolffs, P F G; van den Brule, A J C
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.
Zaria, L T
When tested on solid media the growth of 19 Dermatophilus congolensis strains was inhibited by antibiotic-producing staphylococci isolated from pigs. Two strains, D congolensis D11 and D15, which were very sensitive to the producers and caused lesions of dermatophilosis in a mouse model, were further used to investigate the ability of the producers to inhibit lesion formation by the strains of D congolensis. The simultaneous application of the antibiotic-producing staphylococci and D congolensis suppressed formation of the lesions in the mouse.
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.
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.
McClure, Jo-Ann; Zaal DeLongchamp, Johanna; Conly, John M; Zhang, Kunyan
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.
Huber, Helen; Ziegler, Dominik; Pflüger, Valentin; Vogel, Guido; Zweifel, Claudio; Stephan, Roger
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. 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). 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.
Brilene, Tatjana; Soeorg, Hiie; Kiis, Merilin; Sepp, Epp; Kõljalg, Siiri; Lõivukene, Krista; Jürna-Ellam, Marika; Kalinina, Jelena; Stšepetova, Jelena; Metsvaht, Tuuli; Lutsar, Irja
Coagulase-negative staphylococci (CoNS) are the leading cause of late-onset sepsis (LOS) in neonates. Increasing resistance of CoNS to beta-lactams and aminoglycosides has led to widespread use of vancomycin, which in turn may lead to resistance to vancomycin. Thus, combination therapy of LOS has been advocated. We aimed to determine the interaction of oxacillin and gentamicin against CoNS. In 2005, 34 isolates of oxacillin- and gentamicin-resistant CoNS were obtained from blood samples of neonates with LOS. Combination effect was tested using the checkerboard method, E-test with the other antibiotic incorporated in the medium (E-test-1) and two E-test strips placed in a cross-formation (E-test-2). Of 34 isolates 61.8%, 53% and 73.5% revealed synergy or an additive effect when tested by the checkerboard method, E-test-1 and E-test-2, respectively. Results of all three tests were concordant for six (17.6%) isolates, four showing synergy, and two indifference. Our in vitro results support that combination therapy with penicillinase-resistant penicillin and aminoglycoside can be an alternative to vancomycin. © 2013 APMIS. Published by John Wiley & Sons Ltd.
S. Pehlivanlar Onen
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.
Michel, Astrid; Syring, Claudia; Steiner, Adrian; Graber, Hans U
The association between the contagious Staphylococcus aureus genotype B (GTB) and the presence of coagulase-negative staphylococci (CNS) and Streptococcus spp. (non-agalactiae streptococci), was investigated, and the identification of problem herds without genotyping was evaluated. Milk samples from 10 herds with Staph. aureus GTB herd problems (PH cases) were compared with samples from 19 herds with at least one Staph. aureus isolate of non-B genotype (CH cases). All samples were bacteriologically analysed and Staph. aureus genotyping carried out using a ribosomal spacer-PCR. Cow and quarter prevalences of Staph. aureus, CNS and Streptococcus spp. differed significantly between PH and CH groups. PH cases were highly associated with decreased cow prevalences of CNS and Streptococcus spp. These altered prevalences also contributed significantly to the identification of problem herds without resorting to genotyping. Common herd-level risk factors did not explain the difference between the prevalences in PH and CH cases. Copyright © 2010 Elsevier Ltd. All rights reserved.
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.
De Visscher, A; Piepers, S; Supré, K; Haesebrouck, F; De Vliegher, S
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, Dairy Science Association. Published by Elsevier Inc. All rights reserved.
Cameron, M; Perry, J; Middleton, J R; Chaffer, M; Lewis, J; Keefe, G P
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.
Ma, Xiao Xue; Wang, En Hua; Liu, Yong; Luo, En Jie
Coagulase-negative staphylococci (CoNS) have become increasingly recognized as important agents of nosocomial infection. One of the characteristics of CoNS is their resistance to multiple antimicrobial agents commonly used for the treatment of staphylococcal infections. CoNS strains (n = 745) isolated from a university teaching hospital in China between 2004 and 2009 were tested for antibiotic resistance. The antibiotics were placed into three categories based on resistance levels of the CoNS strains to these antibiotics: high resistance (resistance rate >70 %), including penicillin G, oxacillin and erythromycin; medium resistance (resistance rate between 30 and 70 %), including tetracycline, clindamycin, ciprofloxacin, trimethoprim/sulfamethoxazole and chloramphenicol; and low resistance (resistance rate teicoplanin increased from 4.5 to 6.7 % between 2008 and 2009. A one-step vancomycin agar selection experiment and subsequent population analysis revealed potentially vancomycin-resistant subpopulations that have been selected from the teicoplanin-non-susceptible strains. Vigilant surveillance of nosocomial isolates of CoNS is needed to determine their resistance to glycopeptides.
Cerca, Nuno; Martins, Silvia; Cerca, Filipe; Jefferson, Kimberly K; Pier, Gerald B; Oliveira, Rosário; Azeredo, Joana
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.
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
Fontes, Cláudia Oliveira; Silva, Vânia Lúcia; de Paiva, Mayara Rodrigues Brandão; Garcia, Rafaela Alvim; Resende, Juliana Alves; Ferreira-Machado, Alessandra Barbosa; Diniz, Cláudio Galuppo
Coagulase-negative staphylococci (CoNS), which are generally neglected as foodborne bacteria, are emerging as significant opportunistic pathogens that may be highly resistant to available antimicrobial drugs. In this study, antimicrobial susceptibility patterns, mecA gene occurrence, and virulence-associated characteristics were evaluated in CoNS isolated from soft cheese in Brazil. A total of 227 bacterial isolates were recovered from 35 cheese samples belonging to 5 batches with 7 different trademarks. The CoNS counts ranged from 10(6) to 10(7) CFU/g. High antimicrobial resistance percentages were observed for oxacillin (76.2%), penicillin (78.5%), erythromycin (67.8%), gentamicin (47.2%), clindamycin (35.7%), rifampicin (26.8%), azithromycin (14.7%), tetracycline (14.7%), levofloxacin (14.2%), and sulfamethoxazole-trimethoprim (11.9%). A low antimicrobial resistance percentage was observed for chloramphenicol (2.3%), and all of the tested bacteria were susceptible to vancomycin and linezolid. In total, a multiple antibiotic resistance (MAR) index of >0.2 was observed for 80.6% of the isolated CoNS. However, the MAR index ranged from 50% to 92.6% when only bacterial cheese isolates belonging to the same trademark were considered. Regarding to the prevalence of CoNS carrying mecA gene, 81.5% of the isolated strains were mecA(+) , and 76.2% of these were phenotypically resistant to oxacillin. Three isolates carried the enterotoxin A gene (sea), 29.5% produced biofilm in a laboratory test, and α- or ß-hemolysis were observed for 3% and 5.2%, respectively. This study highlights the extent of the antimicrobial resistance phenomenon in neglected foodborne microorganisms and the potential public health risks that are related to the consumption of CoNS-contaminated soft cheese. © 2013 Institute of Food Technologists®
Lok Bahadur Shrestha
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.
Szczuka, Ewa; Jabłońska, Lucyna; Kaznowski, Adam
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.
Sampimon, O C; Zadoks, R N; De Vliegher, S; Supré, K; Haesebrouck, F; Barkema, H W; Sol, J; Lam, T J G M
In this study, the accuracy of two phenotypic tests, API Staph ID 32 and Staph-Zym, was determined for identification of coagulase-negative staphylococci (CNS) from bovine milk samples in comparison with identification based on DNA-sequencing. A total of 172 CNS isolated from bovine milk were classified into 17 species. The most frequently isolated species based on rpoB sequencing were Staphylococcus chromogenes and Staphylococcus epidermidis, followed by Staphylococcus xylosus, Staphylococcus warneri and Staphylococcus equorum (37, 13, 9, 8 and 6% of isolates, respectively). The API Staph ID 32 correctly identified 41% of the CNS isolates. Best agreement with rpoB sequence based species identification was found for S. epidermidis, Staphylococcus hyicus and S. xylosus (100, 89 and 87%, respectively). The positive predictive value was 89, 100 and 52%, respectively. Poor sensitivity was observed for 3 of the 5 most frequently found species, S. chromogenes (37%), Staphylococcus warneri (15%) and S. equorum (0%) albeit with specificity of 100%. The Staph-Zym needed additional tests for 66% of the isolates and identified 31% of the CNS isolates correctly. Good sensitivity was found for S. epidermidis, S. simulans and S. xyloxus (100, 78 and 73%, respectively). The positive predictive value was 89, 78 and 98%, respectively. Poor sensitivity was observed for S. chromogenes, S. warneri and S. equorum (0, 54 and 0%, respectively) but with a specificity of 100, 99 and 100%, respectively. Both phenotypic tests misidentified a large proportion of CNS isolates and were thus unsuitable for identification of CNS species from bovine milk samples.
Evaluation of BD MAX Staph SR Assay for Differentiating Between Staphylococcus aureus and Coagulase-Negative Staphylococci and Determining Methicillin Resistance Directly From Positive Blood Cultures.
Lee, Jaewoong; Park, Yeon Joon; Park, Dong Jin; Park, Kang Gyun; Lee, Hae Kyung
We evaluated the performance of the BD MAX StaphSR Assay (SR assay; BD, USA) for direct detection of Staphylococcus aureus and methicillin resistance not only in S. aureus but also in coagulase-negative Staphylococci (CNS) from positive blood cultures. From 228 blood culture bottles, 103 S. aureus [45 methicillin-resistant S. aureus (MRSA), 55 methicillin-susceptible S. aureus (MSSA), 3 mixed infections (1 MRSA+Enterococcus faecalis, 1 MSSA+MRCNS, 1 MSSA+MSCNS)], and 125 CNS (102 MRCNS, 23 MSCNS) were identified by Vitek 2. For further analysis, we obtained the cycle threshold (Ct) values from the BD MAX system software to determine an appropriate cutoff value. For discrepancy analysis, conventional mecA/mecC PCR and oxacillin minimum inhibitory concentrations (MICs) were determined. Compared to Vitek 2, the SR assay identified all 103 S. aureus isolates correctly but failed to detect methicillin resistance in three MRSA isolates. All 55 MSSA isolates were correctly identified by the SR assay. In the concordant cases, the highest Ct values for nuc, mecA, and mec right-extremity junction (MREJ) were 25.6, 22, and 22.2, respectively. Therefore, we selected Ct values from 0-27 as a range of positivity, and applying this cutoff, the sensitivity/specificity of the SR assay were 100%/100% for detecting S. aureus, and 97.9%/98.1% and 99.0%/95.8% for detecting methicillin resistance in S. aureus and CNS, respectively. We propose a Ct cutoff value for nuc/mec assay without considering MREJ because mixed cultures of MSSA and MRCNS were very rare (0.4%) in the positive blood cultures.
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.
Prevalence and genetic diversity of arginine catabolic mobile element (ACME) in clinical isolates of coagulase-negative staphylococci: identification of ACME type I variants in Staphylococcus epidermidis.
Onishi, Mayumi; Urushibara, Noriko; Kawaguchiya, Mitsuyo; Ghosh, Souvik; Shinagawa, Masaaki; Watanabe, Naoki; Kobayashi, Nobumichi
Arginine catabolic mobile element (ACME), a genomic island consisting of the arc and/or opp3 gene clusters found in staphylococcal species, is related to increased bacterial adaptability to hosts. Staphylococcus epidermidis is considered a major ACME reservoir; however, prevalence and genetic diversity of ACME in coagulase-negative staphylococci (CNS) have not yet been well characterized for clinical isolates in Japan. A total of 271 clinical isolates of CNS in a Japanese hospital were investigated for the presence and genotype of ACME and SCCmec. The prevalence of ACME-arcA was significantly higher (pepidermidis (45.8%) than in other CNS species (3.7%). ACME in S. epidermidis isolates (n=87) were differentiated into type I (n=33), variant forms of type I (ΔI, n=26) newly identified in this study, type II (n=6), and type ΔII (n=19). ACME-type ΔI, which were further classified into three subtypes, lacked some genetic components between the arc and opp3 clusters in archetypal type I, whereas the arc and opp3 clusters were intact. The arc cluster exhibited high sequence identity (95.8-100%) to that of type I ACME; in contrast, the opp3 cluster was highly diverse, and showed relatively lower identities (94.8-98.7%) to the identical regions in type I ACME. Twenty-one isolates of ΔI ACME-carrying S. epidermidis possessed SCCmec IVa and belonged to ST5 (clonal complex 2). Phylogenetic analysis revealed that isolates harboring ACME ΔI in this study clustered with previously reported S. epidermidis strains with other lineges, suggesting that S. epidermidis originally had some genetic variations in the opp3 cluster. In summary, ACME type ΔI, a truncated variant of ACME-I, was first identified in S. epidermidis, and revealed to be prevalent in ST5 MRSE clinical isolates with SCCmec IVa. Copyright © 2013 Elsevier B.V. All rights reserved.
Maria de Lourdes Ribeiro de Souza da Cunha
Full Text Available Staphylococcal food poisoning is caused by ingestion of enterotoxins preformed in the food contaminated essentially through human manipulation or raw material obtained from animals. Although coagulase-positive Staphylococcus aureus is the main agent responsible for food intoxication, some researches emphasise that coagulase-negative staphylococci (CNS are able to produce staphylococcal enterotoxins and may be a potential cause of food poisoning. In the present study CNS were isolated from foods and the toxigenic capacity of the strains determined. A total of 88 food samples were analysed and 22.7% were positive for CNS strains. Staphylococcal counts ranged from 3.0 x 10² to 1.4 x 10(6 CFU/g or mL of food examined. S. epidermidis predominated among the isolates (40%. Further isolates included S. xylosus (20%, S. warneri (20%, S. saccharolyticus (15%, and S. hominis (5%. Four isolates were positive for enterotoxin genes, as detected by polymerase chain reaction, with sea being the predominant gene. Although no enterotoxin production was detected by the reverse passive latex agglutination method, the data showed that the toxigenic capacity of CNS should not be ignored, requiring investigation of this group of microorganisms in food.A intoxicação alimentar estafilocócica ocorre devido à ingestão de alimentos contaminados com enterotoxinas. Essa contaminação tem sido oriunda, principalmente, da manipulação humana, ou de matérias-primas procedentes de animais portadores. Embora Staphylococcus aureus coagulase positiva, seja o principal agente de intoxicação alimentar, alguns pesquisadores enfatizam que os estafilococos coagulase-negativa (ECN podem produzir as enterotoxinas estafilocócicas, podendo contribuir para a intoxicação alimentar. Este estudo teve como objetivos isolar os ECN de alimentos e verificar a capacidade enterotoxigênica dessas linhagens. Foram estudadas 88 amostras de alimentos, sendo que 22,7% foram positivas para
Boamah, Vivian Etsiapa; Agyare, Christian; Odoi, Hayford
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...... (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......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...
Qiao, B; Cui, J-Y; Sun, L; Yang, S; Zhao, Y-L
To develop and evaluate cross-priming amplification (CPA) combined with immuno-blotting for the detection of coagulase-positive Staphylococci including Staphylococcus aureus. Twenty-four sets of cross and detection primers were designed according to four sequences of coagulase gene in Staph. aureus. The most specific primer pair was screened out for the next amplification and interaction. The specificity was evaluated in a total of 53 species of Staph. aureus and non-Staph. aureus. Two red lines indicating positive were always observed on the BioHelix Express strip for 12 subspecies of Staph. aureus. In contrast, only one signal line showing negative results was detected in all of non-Staph. aureus samples. The limit of detection (LOD) of CPA was 3·6 ± 2·7 fg for the genomic DNA, which is about 100 and 10 times sensitive than those of PCR and loop-mediated isothermal amplification respectively. For the pure culture of Staph. aureus and milk powders, the LODs of CPA were about 1·34 CFU per reaction and 5·2 ± 3·7 CFU per 100 g of milk powder respectively. The CPA method was also successfully applied to evaluate the contamination of Staph. aureus in 318 samples of daily food. CPA is a very sensitive and rapid method to detect Staph. aureus by simple laboratory instrument. It is the first report on the application of the CPA with immuno-blotting for detection of coagulase-positive Staphylococci including Staph. aureus. © 2015 The Society for Applied Microbiology.
Cameron, M; Barkema, H W; De Buck, J; De Vliegher, S; Chaffer, M; Lewis, J; Keefe, G P
This study evaluated matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS) for the identification of bovine-associated coagulase-negative staphylococci (CNS), a heterogeneous group of different species. Additionally, we aimed to expand the MALDI-ToF MS database with new reference spectra as required to fill the gaps within the existing commercial spectral library. A total of 258 isolates of CNS were used in the study, covering 16 different CNS species. The majority of the isolates were previously identified by rpoB gene sequencing (n = 219), and the remainder were identified by sequencing of 16S rRNA, hsp60, or both rpoB and hsp60. The genotypic identification was considered the gold standard identification. All MALDI-ToF MS identifications were carried out using the direct transfer method. In a preliminary evaluation (n = 32 isolates; 2 of each species) with the existing commercial database, MALDI-ToF MS showed a typeability of 81% (26/32) and an accuracy of 96% (25/26). In the main evaluation (n = 226 isolates), MALDI-ToF MS with the existing commercial Biotyper (Bruker Daltonics Inc., Billerica, MA) database achieved a typeability of 92.0% (208/226) and an accuracy of 99.5% (207/208). Based on the assessment of the existing commercial database and prior knowledge of the species, a total of 13 custom reference spectra, covering 8 species, were created and added to the commercial database. Using the custom reference spectra expanded database, isolates were identified by MALDI-ToF MS with 100% typeability and 100% accuracy. Whereas the MALDI-ToF MS manufacturer's cutoff for species-level identification is 2.000, the reduction of the species level cutpoint to ≥1.700 improved the species-level identification rates (from 64 to 92% for the existing commercial database) when classifying CNS isolates. Overall, MALDI-ToF MS using the direct transfer method was shown to be a highly reliable tool for the identification of bovine
Faghri, Jamshid; Razavi, Mohamad Reza
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
Bochniarz, Mariola; Zdzisińska, Barbara; Wawron, Władysław; Szczubiał, Marek; Dąbrowski, Roman
The aim of the study was to evaluate the concentrations of cytokines IL-4, IL-6, and IL-10 and acute phase protein amyloid A in milk and in serum from cows with subclinical mastitis caused by coagulase-negative staphylococci and from healthy cows. The blood and milk samples were obtained from 35 midlactation, multiparous (between parities 2 and 4) Holstein-Friesian cows. In the milk samples from 20 cows with subclinical mastitis, the following species of Staphylococcus were detected: Staphylococcus xylosus (8 samples), Staphylococcus chromogenes (6 samples), Staphylococcus haemolyticus (2 samples), Staphylococcus simulans (2 samples), and Staphylococcus sciuri (2 samples). The results of the present study indicate that the level of IL-6 in cows suffering from subclinical mastitis tended to be high in both serum and milk (432.09 and 254.32 pg/mL) compared with the level in healthy cows (164.47 and 13.02 pg/mL, respectively). Amyloid A value also was significantly higher in milk of unhealthy cows compared with cows without subclinical mastitis (790.2 and 360.5 ng/mL). No significant differences were found in levels of amyloid A in serum of both tested groups of cows (2,680.0 and 2,720.0 ng/mL). In contrast, concentration of IL-4 was significantly lower both in serum and in milk of cows with staphylococcal mastitis (86.1 and 123.17 pg/mL) compared with control animals (413.5 and 670.2 pg/mL). The level of IL-10 also was significantly higher in milk of healthy cows than in infected cows (39.78 and 22.5 pg/mL); however, differences in serum levels of this cytokine between tested groups were significantly less important (220.6 and 175.1 pg/mL). Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
Identification of ABC transporter genes conferring combined pleuromutilin-lincosamide-streptogramin A resistance in bovine methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci.
Wendlandt, Sarah; Kadlec, Kristina; Feßler, Andrea T; Schwarz, Stefan
The aim of this study was to investigate the genetic basis of combined pleuromutilin-lincosamide-streptogramin A resistance in 26 unrelated methicillin-resistant Staphylococcus aureus (MRSA) and coagulase-negative staphylococci (CoNS) from dairy cows suffering from mastitis. The 26 pleuromutilin-resistant staphylococcal isolates were screened for the presence of the genes vga(A), vga(B), vga(C), vga(E), vga(E) variant, sal(A), vmlR, cfr, lsa(A), lsa(B), lsa(C), and lsa(E) by PCR. None of the 26 isolates carried the genes vga(B), vga(C), vga(E), vga(E) variant, vmlR, cfr, lsa(A), lsa(B), or lsa(C). Two Staphylococcus haemolyticus and single Staphylococcus xylosus, Staphylococcus lentus, and Staphylococcus hominis were vga(A)-positive. Twelve S. aureus, two Staphylococcus warneri, as well as single S. lentus and S. xylosus carried the lsa(E) gene. Moreover, single S. aureus, S. haemolyticus, S. xylosus, and Staphylococcus epidermidis were positive for both genes, vga(A) and lsa(E). The sal(A) gene was found in a single Staphylococcus sciuri. All ABC transporter genes were located in the chromosomal DNA, except for a plasmid-borne vga(A) gene in the S. epidermidis isolate. The genetic environment of the lsa(E)-positive isolates was analyzed using previously described PCR assays. Except for the S. warneri and S. xylosus, all lsa(E)-positive isolates harbored a part of the previously described enterococcal multiresistance gene cluster. This is the first report of the novel lsa(E) gene in the aforementioned bovine CoNS species. This is also the first identification of the sal(A) gene in a S. sciuri from a case of bovine mastitis. Moreover, the sal(A) gene was shown to also confer pleuromutilin resistance. Copyright © 2015 Elsevier B.V. All rights reserved.
Sánchez Mainar, María; Leroy, Frédéric
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
Factors associated with intramammary infection in dairy cows caused by coagulase-negative staphylococci, Staphylococcus aureus, Streptococcus uberis, Streptococcus dysgalactiae, Corynebacterium bovis, or Escherichia coli.
Taponen, S; Liski, E; Heikkilä, A-M; Pyörälä, S
The aim of this study was to determine risk factors for bovine intramammary infection (IMI) associated with the most common bacterial species in Finland. Large databases of the Finnish milk-recording system and results of microbiological analyses of mastitic milk samples from Valio Ltd. (Helsinki, Finland) were analyzed. The study group comprised 29,969 cows with IMI from 4,173 dairy herds. A cow with a quarter milk sample in which DNA of target species was detected in the PathoProof Mastitis PCR Assay (Thermo Fisher Scientific, Waltham, MA) was determined to have IMI. Only cows with IMI caused by the 6 most common pathogens or groups of pathogens, coagulase-negative staphylococci (CNS), Staphylococcus aureus, Streptococcus uberis, Streptococcus dysgalactiae, Corynebacterium bovis, and Escherichia coli, were included. The control group comprised 160,176 IMI-free cows from the same herds as the study group. A multilevel logistic regression model was used to study herd- and cow-specific risk factors for incidence of IMI. Pathogen-specific results confirmed those of earlier studies, specifically that increasing parity increases prevalence of IMI regardless of causative pathogen. Holsteins were more susceptible to IMI than Nordic Reds except when the causative pathogen was CNS. Occurrence of IMI caused by C. bovis was not related to milk yield, in contrast to IMI caused by all other pathogens investigated. Organic milk production was associated with IMI only when the causative pathogen of IMI was Staph. aureus; Staph. aureus IMI was more likely to occur in conventional than in organic production. Cows in older freestall barns with parlor milking had an increased probability of contracting an IMI compared with cows in tiestall barns or in new freestall barns with automatic milking. This was the case for all IMI, except those caused by CNS, the prevalence of which was not associated with the milking system, and IMI caused by Staph. aureus, which was most common in cows
Maria de Lourdes R.S. Cunha
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
Dos Santos, Danielle Cabral; Lange, Carla Christine; Avellar-Costa, Pedro; Dos Santos, Katia Regina Netto; Brito, Maria Aparecida Vasconcelos Paiva; Giambiagi-deMarval, Marcia
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.
Application of Matrix-assisted Laser Desorption Ionization Time-of-flight Mass Spectrometry for Identification of Coagulase-negative Staphylococci Isolated from Milk of Cows with Subclinical Mastitis.
Banach, T; Bochniarz, M; Łyp, P; Adaszek, Ł; Wawron, W; Furmaga, B; Skrzypczak, M; Ziętek, J; Winiarczyk, S
The aim of this study was to use matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of coagulase-negative staphylococci (CNS) isolated from the milk of cows with subclinical mastitis. The study material consisted of 33 isolates of CNS, identified by the results of API Staph tests, obtained from the milk of cows with subclinical mastitis. Based on the spectra analyses, MALDI-TOF MS tests of 33 bacterial samples allowed identification of the microorganisms in 27 cases (81.8%). The most frequent cause of subclinical mastitis was found to be Staphylococcus sciuri (39%), while S. vitulinus was detected in 15% of the milk samples. The results obtained indicate that MALDI-TOF MS can be used for the identification of CNS isolated from bovine mastitis as a method supplementary to biochemical tests.
A multiplex PCR assay for the rapid and sensitive detection of methicillin-resistant Staphylococcus aureus and simultaneous discrimination of Staphylococcus aureus from coagulase-negative staphylococci.
Xu, Benjin; Liu, Ling; Liu, Li; Li, Xinping; Li, Xiaofang; Wang, Xin
Methicillin-resistant Staphylococcus aureus (MRSA) is a global health concern, which had been detected in food and food production animals. Conventional testing for detection of MRSA takes 3 to 5 d to yield complete information of the organism and its antibiotic sensitivity pattern. So, a rapid method is needed to diagnose and treat the MRSA infections. The present study focused on the development of a multiplex PCR assay for the rapid and sensitive detection of MRSA. The assay simultaneously detected 4 genes, namely, 16S rRNA of the Staphylococcus genus, femA of S. aureus, mecA that encodes methicillin resistance, and one internal control. It was rapid and yielded results within 4 h. The analytical sensitivity and specificity of the multiplex PCR assay was evaluated by comparing it with the conventional method. The analytical sensitivity of the multiplex PCR assay at the DNA level was 10 ng DNA. The analytical specificity was evaluated with 10 reference staphylococci strains and was 100%. The diagnostic evaluation of MRSA was carried out using 360 foodborne staphylococci isolates, and showed 99.1% of specificity, 96.4% of sensitivity, 97.5% of positive predictive value, and 97.3% of negative predictive value compared to the conventional method. The inclusion of an internal control in the multiplex PCR assay is important to exclude false-negative cases. This test can be used as an effective diagnostic and surveillance tool to investigate the spread and emergence of MRSA. © 2012 Institute of Food Technologists®
Duration of the natural infection of the mammary gland by coagulase negative staphylococci in primiparous cows /Duração das infecções naturais da glândula mamaria por estafilococos coagulase negativos em vacas primíparas
Juliana Alves Dias
Full Text Available One hundred fourteen primiparous cows wre analized. They wre from nine milk properties of the west region of São Paulo sate. On the seventh day of lactation. 42 (36.84% showed infections in 69 (41.07% quarters. The main aetiologic agents of the intramammary infections on the seventh day were coagulase-negative staphylococci (CMS with 53,62%, followed by Corynebacterium spp with 26r08% and coagulase-positive staphylococci (CPS, with 11,59% of the positive samples. From the seventh day of lactation on there was a decreasing tendency of the infections by SCN in the previously tnfected mammary quarter (r = 0,63, r2= 0,4 and simultaneously there were new infections by other pathogen in these quarter. On the 150th day, the CNS were not isolated. There was not significant difference (P3 0.05 about the number of infections by udder quarter There was significant difference (P£0rO1 between the average of the somatic cell count (SCC of the milk samples. It. comes from the infected quarters by SCN (397 x 103/ mL and of negative quarters (104 x 103/mL.
Taponen, Suvi; Simojoki, Heli; Haveri, Maarit; Larsen, Helle D; Pyörälä, Satu
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-typing of CNS was compared with the phenotypic identification. A total of 133 clinical or subclinical quarter cases of intramammary infection caused by CNS from the practice area of the Ambulatory Clinic of the University of Helsinki were studied. Bacteriological diagnosis was based on biochemical (API) testing. Staphylococcus simulans (43.6%) followed by S. chromogenes (23.3%) were the most common CNS species isolated from the milk samples. Ninety-nine isolates were genotyped using AFLP-analysis. Only 75.0% of S. chromogenes and S. simulans isolates identified with API test were clustered with the type strains 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 beta-lactamase-negative CNS was treated with penicillin G and that due to beta-lactamase-positive CNS with cloxacillin. Nineteen percent of the isolates were beta-lactamase-positive. The bacterial cure rate for quarters treated with antimicrobials was high, 85.9%, as opposed to only 45.5% for untreated quarters. Bacterial cure rates for the most common CNS species or AFLP clusters were not statistically different. Further studies on identification of CNS species are needed.
Veras, Jamaira Fereira; do Carmo, Luiz Simeão; Tong, Lawrence C; Shupp, Jeffrey W; Cummings, Christiano; Dos Santos, Deise Aparecida; Cerqueira, Mônica Maria Oliveira Pinho; Cantini, Alvaro; Nicoli, Jacques Robert; Jett, Marti
The purpose of this study was to identify enterotoxin genes from isolates of coagulase-negative staphylococci and coagulase-positive staphylococci obtained from dairy products, responsible for 16 outbreaks of food poisoning. From the pool of 152 staphylococcal isolates, 15 coagulase-negative and 15 coagulase-positive representatives were selected for this study. The 15 coagulase-negative isolates were tested for the presence of coa and femA genes, which are known to be characteristic of Staphylococcus aureus. After testing for enterotoxin genes by polymerase chain reaction (PCR), the 30 selected isolates were tested for the presence of toxin by immunoassay. Seven of the coagulase-negative isolates amplified the coa gene and were subsequently reclassified as coagulase-positive. Twenty-one of 30 selected isolates had staphylococcal enterotoxin genes and most of these produced toxin as well. The most frequently encountered enterotoxin genes were sea and seb. Among eight coagulase-negative isolates, five had enterotoxin genes, all of which were found to have detectable toxin by immunoassay. The results from this study demonstrate that coagulase-negative as well as coagulase-positive staphylococci isolated from dairy products are capable of genotypic and phenotypic enterotoxigenicity. Furthermore, these data demonstrate that PCR is a sensitive and specific method for screening outbreak isolates regardless of coagulase expression.
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%. .
A comparison of PCR detection of Meca with oxacillin disk susceptibility testing in different media and sceptor automated system for both Staphylococcus aureus and coagulase-negative staphylococci isolates
Full Text Available Purpose: To evaluate three methods for 406 isolates of Staphylococcus aureus and coagulase-negative staphylococci (CNS for the detection of methicillin resistance (MR using National Committee for Clinical Laboratory Standards (NCCLS new interpretive criteria. Methods: We used polymerase chain reaction (PCR as a gold standard method to evaluate three methods [disk diffusion with Mueller-Hinton agar (MHA and mannitol salt agar (MSA and Sceptor system (Becton Dickinson, USA] for the detection of mecA gene. The isolates that were methicillin-resistant with any of the three tests were evaluated further for MR by E-test. Results: MHA, MSA and Sceptor showed sensitivities of 100, 100 and 99% for S. aureus and 100, 82.6 and 72.1% for CNS, respectively. The specificities of the same methods were found as 100, 90.1 and 99.3% for S. aureus and 79.2, 95.8 and 97.2% for CNS, respectively. E-test showed 100% sensitivity for both S. aureus and CNS. Forty-eight CNS and 16 S. aureus isolates, which presented discrepancies with the three phenotypic methods (MHA disk diffusion method, MSA disk diffusion method and Sceptor, were correctly classified as resistant/susceptible with the E-test when compared with PCR. Only five CNS isolates, which were mecA-negative with PCR were resistant with E-test. Analysis of 248 S. aureus revealed that MHA is superior to other phenotype-based susceptibility testing methods in detecting MR. When we examined the results of 158 CNS, none of the three methods proved efficient in detecting MR. Conclusions: We conclude that although the accuracy of the MHA disk diffusion test for the detection of MR approaches the accuracy of PCR for S. aureus isolates, the need for easy and reliable methods of detecting MR in CNS still remains.
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
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.
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
Maria de Lourdes Ribeiro de Souza da Cunha
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
Comparison of growth on mannitol salt agar, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, VITEK® 2 with partial sequencing of 16S rRNA gene for identification of coagulase-negative staphylococci.
Ayeni, Funmilola A; Andersen, Camilla; Nørskov-Lauritsen, Niels
Mannitol salt agar (MSA) is often used in resources' limited laboratories for identification of S. aureus however, coagulase-negative staphylococci (CoNS) grows and ferments mannitol on MSA. 171 strains of CoNS which have been previously misidentified as S. aureus due to growth on MSA were collected from different locations in Nigeria and two methods for identification of CoNS were compared i.e. ViTEK 2 and MALDI-TOF MS with partial 16S rRNA gene sequencing as gold standard. Partial tuf gene sequencing was used for contradicting identification. All 171 strains (13 species) grew on MSA and ferments mannitol. All tested strains of S. epidermidis, S. haemolyticus, S. nepalensis, S. pasteuri, S. sciuri,, S. warneri, S. xylosus, S. capitis were correctly identified by MALDI-TOF while variable identification were observed in S. saprophyticus and S. cohnii (90%, 81%). There was low identification of S. arlettae (14%) while all strains of S. kloosii and S. gallinarum were misidentified. There is absence of S. gallinarum in the MALDI-TOF database at the period of this study. All tested strains of S. epidermidis, S. gallinarum, S. haemolyticus, S. sciuri,, S. warneri, S. xylosus and S. capitis were correctly identified by ViTEK while variable identification were observed in S. saprophyticus, S. arlettae, S. cohnii, S. kloosii, (84%, 86%, 75%, 60%) and misidentification of S. nepalensis, S. pasteuri. Partial sequencing of 16S rRNA gene was used as gold standard for most strains except S. capitis and S. xylosus where the two species were misidentified by partial sequencing of 16S rRNA contrary to MALDI-TOF and ViTEK identification. Tuf gene sequencing was used for correct identification. Characteristic growth on MSA for CoNS is also identical to S. aureus growth on the media and therefore, MSA could not differentiate between S. aureus and CoNS. The percentage accuracy of ViTEK was better than MALDI-TOF in identification of CoNS. Although partial sequencing of
Nosocomial sepsis is a major cause of morbidity in the neonatal intensive care unit (NICU), with coagulase-negative staphylococci (CONS) generally reported to be the most frequent causative micro-organisms. There is substantial evidence for the association between CONS sepsis and indwelling
Crisley, F. D.; Peeler, J. T.; Angelotti, R.
Five selective media for the detection and enumeration of coagulase-positive staphylococci were evaluated for their efficiency in the recovery of 17 strains of coagulase-positive staphylococci from foods. They were Staphylococcus Medium 110 (SM-110), tellurite-glycine-agar (TGA), egg-tellurite-glycine-pyruvate-agar (ETGPA), tellurite-egg-agar (TEA), and tellurite-polymyxin-egg yolk-agar (TPEY). Statistical analysis by the rank correlation method of the efficiency with which these media recovered staphylococci from pure 24-hr Brain Heart Infusion cultures revealed the following efficiencies in descending order: (i) TPEY, (ii) ETGPA, (iii) TGA, (iv) TEA, (v) SM-110. Growth of 17 strains of coagulase-negative cocci on these media showed the following approximate descending order of inhibition to these organisms: (i) ETGPA, (ii) TEA, (iii) SM-110, (iv) TGA, (v) TPEY. The appearance of colonies of the various coagulase-negative strains on each medium was studied for the degree to which they could be confused with colonies of coagulase-positive strains. Nineteen food contaminants, including Proteus vulgaris, Bacillus sp., Escherichia coli, Erwinia sp., fecal streptococci, and others, were also studied for similarities in appearance to staphylococci and for ability to grow on the selective media. The influence of five sterile food homogenates (frozen chicken and tuna pies, custard, smoked ham, and raw whole egg) on recovery of 1,500 enterotoxigenic staphylococci (three strains) per milliliter was determined by statistical analysis. Three main effects (culture, media, and food) and three interactions (media with food, food with cultures, and media with culture) were found to be significant. Recovery on TPEY was influenced less by food than the other selective media and showed optimal recovery ability from sterile custard, eggs, and ham. TGA recovered well from sterile chicken pie and custard, SM-110 from sterile custard, and TEA from sterile ham. None of the media was
Winstel, Volker; Kühner, Petra; Rohde, Holger; Peschel, Andreas
Coagulase-negative staphylococci (CoNS) are recognized as significant opportunistic pathogens. However, current knowledge of virulence mechanisms is very limited because a significant proportion of CoNS are refractory to available techniques for DNA transformation. We describe an efficient protocol for plasmid transfer using bacteriophage Φ187, which can transduce plasmid DNA to a wide range of CoNS from a unique, engineered Staphylococcus aureus strain. The use of a restriction-deficient, modification-proficient S. aureus PS187 mutant, which has a CoNS-type bacteriophage surface receptor, allows plasmid transfer to CoNS even when they are refractory to electroporation. Once the Φ187 titer reaches 10(9) plaque-forming units per milliliter, plasmid transfer can be accomplished within 1-2 d. Thus, our protocol is a major technical advance offering attractive opportunities for research on CoNS-mediated infections.
Nikolina Velizarova Rusenova
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.
Staphylococci: colonies morphological caracteristics, coagulase and EEA production colected from cooled raw milk samples/ Estafilococos: morfologia das colônias, produção de coagulase e enterotoxina a, em amostras isoladas de leite cru refrigerado
Wladimir P. Silva
Full Text Available The stories of food poisoning from staphylococci, mainly by S. aureus, are frequently associates to the ingestion of food with 100 ng of enterotoxin/g or mL. The production of enterotoxin using staphylococci is strongly associated with the production of coagulase. In this paper, milk samples of 51 milk properties in Londrina (RS and 50 in Pelotas had been evaluated. All the 101 samples had presented counts of coagulase positive staphylococci (CPS. From 552 colonies studied, 256 were CPS where 60% of the colonies were typical in Baird Parker agar and, from 296 coagulase negative staphylococcus (CNS, in mean, 60% were atypical colonies. Due to the enterotoxigenic capacity, were tested 109 CPS, being 5,5% (6 positive and 18 CNS where 1 (5,5% were positive. The frequency of CPS EEA (5,5% producer was similar of the CNS. There is a wide milk contamination with CPS, where, a big part (40% of the atypical colonies is coagulase producers.Os relatos de intoxicação estafilocócica, principalmente por S. aureus, são frequentemente associados à ingestão de 100 ng de enterotoxina/g ou mL de alimento. A produção de enterotoxina pelos estafilococos está associada fortemente à produção de coagulase. Neste trabalho foram avaliadas amostras de leite de 51 propriedades leiteiras em Londrina (RS e 50 em Pelotas. Todas as 101 amostras apresentaram contagens de estafilococos coagulase positivos (ECP. Das 552 colônias avaliadas, 256 foram coagulase positivas e destas cerca de 60% eram típicas no àgar Baird Parker e, em média, 60% do total de 296 estafilococos coagulase negativos (ECN eram colônias atípicas. Quanto à capacidade enterotoxigênica, testou-se 109 colônias ECP, sendo 5,5% (6 positivas e 18 ECN, sendo 1 (5,5% positiva. Assim, a freqüência de ECN produtores de EEA (5,5% foi semelhante a dos ECP. Conclui-se que há extensa contaminação do leite cru por ECP, sendo grande parte (40% das colônias atípicas produtoras de coagulase.
Krasniqi, Xhevdet; Rexhepi, Sylejman; Gashi, Masar; Berisha, Blerim; Abazi, Flora; Koçinaj, Dardan
We report a case of a 49-year-old patient who developed poststaphylococcal coagulase negative reactive arthritis. The woman presented with constitutional symptoms, arthritis, urinary infection and conjunctivitis. The blood culture was positive for the staphylococcal coagulase negative infection. Erythrocyte sedimentation rate and C-reactive protein were elevated, whereas the rheumatoid factor was negative. Radiographic findings confirmed diagnosis of pleuropneumonia, and one year later of chronic asymmetric sacroileitis. Physicians should be aware of possible reactive arthritis after staphylococcal coagulase negative bacteremia.
Martins, Paula Dalcin; de Almeida, Taiana Trindade; Basso, Ana Paula; de Moura, Tiane Martin; Frazzon, Jeverson; Tondo, Eduardo César; Frazzon, Ana Paula Guedes
Coagulase-positive staphylococci (CPS) cause staphylococcal food poisoning. Recently, these bacteria have received increasing attention due to their potential role in the dissemination of antibiotic resistance markers. The present study aimed to evaluate coagulase-positive staphylococci counts, species distribution, enterotoxin genes prevalence, and the antibiotic resistance profile of CPS isolated from in natura chicken meat. Fifteen frozen and 15 chilled industrialized, uncooked chicken parts or entire carcasses were used. Staphylococcal counts revealed that frozen chicken meat samples displayed the lowest CPS count compared with chilled chicken meat samples (pStaphylococcus aureus (62%) was the most common species, followed by S. intermedius, S. delphini, and S. schleiferi subsp. coagulans (10% each) and S. hyicus (8%). The polymerase chain reaction identification of sea, seb, sec, sed, and see genes revealed that 70% of the isolates harbored at least one enterotoxin gene, with sea and sed being the most frequently encountered ones. Two of the 50 investigated strains harbored three different enterotoxin genes. A high frequency of isolates resistant to penicillin, teicoplanin, oxacillin, and clindamycin was observed, and 80% of CPS were found to be resistant to at least one of the 11 tested antimicrobials. Vancomycin-resistant S. aureus and S. intermedius showed minimum inhibitory concentrations of 512 and 64 μg/mL, respectively. These isolates might indicate the dissemination of vancomycin resistance in the community and imply food safety hazards.
Effects of intra-mammary bacterial infection with coagulase negative staphylococci and stage of lactation on shedding of epithelial cells and infiltration of leukocytes into milk: comparison among cows, goats and sheep.
Leitner, Gabriel; Merin, Uzi; Krifucks, Oleg; Blum, Shlomo; Rivas, Ariel L; Silanikove, Nissim
The effects of mammary gland bacterial infection and stage of lactation on leukocyte infiltration into the mammary gland were compared among cows, goats and sheep. Animals were at two stages of lactation: mid or late. In mid-lactation animals, bacterial-free glands and coagulase negative Staphylococcus (CNS)-infected glands were compared. In late lactation only uninfected glands were studied. Of mid-lactation bacteria-free animals, goats had the highest number of leukocytes and % polymorphonuclears (PMNs), whereas sheep had the lowest and leukocytes number in cows were intermediate between sheep and goats. Based on %PMN, two cell clusters were found in sheep, which overlapped with the parallel cell clusters of cows and goats, but with a slightly higher number of leukocytes in each cell cluster. At late lactation, goats had higher values for %PMN and leukocyte numbers in comparison to cows, which had a similar cellular profile to sheep. The cellular immune response to CNS infection was similar for the three animal species, although the number of cells was different, while the basal cell level at mid-lactation and especially at the end of lactation was species specific. Copyright © 2012 Elsevier B.V. All rights reserved.
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
Lucía E. Alcaráz
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
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
Adália Dias Dourado Oliveira
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
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.
Krasniqi, Xhevdet; Rexhepi, Sylejman; Gashi, Masar; Berisha, Blerim; Abazi, Flora; Ko?inaj, Dardan
We report a case of a 49-year-old patient who developed poststaphylococcal coagulase negative reactive arthritis. The woman presented with constitutional symptoms, arthritis, urinary infection and conjunctivitis. The blood culture was positive for the staphylococcal coagulase negative infection. Erythrocyte sedimentation rate and C-reactive protein were elevated, whereas the rheumatoid factor was negative. Radiographic findings confirmed diagnosis of pleuropneumonia, and one year later of chr...
López-Fabal, Fátima; Román Alonso, Federico; Almagro Moltó, María; Sanz Rodríguez, Nuria; Gómez-Garcés, José Luis
We recovered 22 coagulase-negative staphylococci isolates in our hospital to study their identity, susceptibility, epidemiological profile, linezolid resistance mechanisms, and the possibilities of different antibiotic combinations. Isolate identification was performed using mass spectrometry (Vitek-MS, bioMérieux). Susceptibility testing was carried out with the Vitek-2 system and the broth microdilution method according to CLSI guidelines. Pulsed-field gel electrophoresis (PFGE) was performed to analyze the genetic relationship between isolates. Linezolid resistance mechanisms were evaluated by PCR/sequencing: presence of cfr gene, point mutations in domain V of 23S ribosomal RNA and additional ribosomal mutations (in the rplC, rplD and rplV genes). The in vitro activity of linezolid was investigated alone and in combination with another three antibiotics acting on different cellular targets, using E-test strips. Twenty isolates were identified as Staphylococcus epidermidis, and 2 as Staphylococcus hominis. PFGE showed that isolates belonged to diverse clones, 21 of them presented mutations in the domain V region of 23S rRNA and the cfr gene was found in 54.5%. Prior administration of linezolid was documented in most of cases. Linezolid in combination with gentamicin showed a synergistic activity in 45.5% of isolates. Staphylococcus epidermidis was the most prevalent linezolid-resistant coagulase-negative staphylococci. All isolates showed increased MIC values compared to other anti-staphylococcal drugs and several linezolid resistance mechanisms. Our data suggest that linezolid plus gentamicin could be a synergistic combination against linezolid-resistant coagulase-negative staphylococci. Copyright © 2012 Elsevier España, S.L. All rights reserved.
Normanno, G; Firinu, A; Virgilio, S; Mula, G; Dambrosio, A; Poggiu, A; Decastelli, L; Mioni, R; Scuota, S; Bolzoni, G; Di Giannatale, E; Salinetti, A P; La Salandra, G; Bartoli, M; Zuccon, F; Pirino, T; Sias, S; Parisi, A; Quaglia, N C; Celano, G V
Staphylococcus aureus is a very common organism capable of producing several enterotoxins (SEs) that cause intoxication symptoms of varying intensity in humans when ingested through contaminated food. This paper reports the results of an investigation on the presence of Coagulase-Positive Staphylococci (CPS) and S. aureus in several food products marketed in Italy and on food contact surface swabs sampled from the food industry. A total of 11,384 samples were examined and 1971 of them (17.3%) were found to contain CPS. The assays performed on 541 CPS strains led to the identification of 537 S. aureus strains on which characterization of type A, B, C and D staphylococcal enterotoxins (SEA, SEB, SEC and SED) was performed. A total of 298 S. aureus strains (55.5%) produced one or more SEs: 33.9% of the strains produced SEC, 26.5% SEA, 20.5% SEA+SED, 13.4% SED, 2.7% SEB, 1.7% SEA+SEB, 0.7% SEC+SED and 0.3% produced SEA+SEC and SEB+SEC. The investigation highlighted that these organisms are very common and constitute a potential risk for consumers' health.
Schwarz, Stefan; Alesík, Eva; Werckenthin, Christiane; Grobbel, Mirjam; Lübke-Becker, Antina; Wieler, Lothar H; Wallmann, Jürgen
A total of 248 coagulase-positive and coagulase-variable staphylococci from two indications of swine (infections of the urinary/genital tract including strains from the mastitis metritis agalactia syndrome as well as infections of the skin) as well as two indications of dogs/cats (respiratory tract infections and infections of skin/ear/mouth) were investigated for their susceptibility to numerous antimicrobial agents. Regardless of the animal origin and indication, the most frequently detected resistance properties were resistances against penicillin G (53-77%) and ampicillin (42-75%), tetracycline (33-52%) as well as erythromycin (13-27%). Oxacillin-resistant staphylococci were rarely detected.
Bogiel, Tomasz; Mikucka, Agnieszka; Deptuła, Aleksander; Gospodarek, Eugenia
Many identification and relatedness studies methods had been commonly used for epidemiological studies in microbiological laboratories. Apart from phenotypic methods, genotypic are also often used. The aim of this study was to compare, obtained by PFGE chromosomal DNA patterns of methicillin-resistant S. epidermidis strains isolated from clinical material. 46 methicillin-resistant S. epidermidis strains were included in this study. Most of them were isolated from wound swabs (65.2%) and catheters (19.6%) from different surgical clinics (76.1%). To identify strains and receive biochemical profiles, ID 32 Staph tests and GPI cards of Vitek 1 system were used. Pulsed-field gel electrophoresis and Tenover et al. interpretation were used to compare chromosomal DNA patterns of examined strains. 44 and 42 PFGE patterns of chromosomal DNA were received, using visual interpretation classifying two pairs of strains as the same, two pairs as closely related and three pairs as probably related. Strains classified as identical and similar in visual evaluation were indistinguishable in Molecular Analyst DST interpretation, probably due to tolerance in bands location pattern. Strains probably related in visual interpretation represent at least 96% similarity in Molecular Analyst DST but different susceptibility and biochemical profiles obtained by ID 32 Staph and Vitek 1. PFGE analysis had foremost capacity to distinguish methicillin-resistant S. epidermidis strains using visual interpretation and Molecular Analyst DST (Bio-Rad) program and seems to be useful method in epidemiological studies. Strains with the same PFGE pattern, had different susceptibility and biochemical profiles.
Full Text Available Microbiological analyses of chicken eggs in Recife and Salvador have shown a high occurrence of Salmonella in the egg shells and yolks. Likewise, the occurrence of Salmonella plus coagulase-positive staphylococci in Coalho cheese reached alarming levels. The data revealed a significant risk of infections and intoxications from consuming these foods in the cities.
Moura, Tiane Martin de; Campos, Fabrício Souza; d'Azevedo, Pedro Alves; Van Der Sand, Sueli Teresinha; Franco, Ana Cláudia; Frazzon, Jeverson; Frazzon, Ana Paula Guedes
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. 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. 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%). 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.
Tiane Martin de Moura
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.
Oliveira Lis Christina de
Full Text Available This study evaluated the middle ear microbiota and antimicrobial susceptibility patterns from strains isolated from dogs with otitis media. A total of 62 dogs obtained from Zoonoses Control Center of Fortaleza City Ceará State / Brazil were studied over a 10-month period (August/2003 to June/2004. Of the total, 46.8% (n=30 of the animals were positive for otitis media and the infection was monomicrobial in 76.6% of them. The most frequent isolated agents were coagulase-positive Staphylococci (CPS-55% and Pseudomonas sp (10%. For S. intermedius (n=13 and S. aureus subsp aureus (n=9, respectively, the greater resistance rates were observed using penicillin G (30.76% and 44.44%, ampicillin (7.69% and 44.44%, erythromycin (23.07% and 44.44%, clindamycin (23.07% and 44.44% and thrimethropim/sulfamethoxazol (15.38% and 33.33%.
Velázquez-Guadarrama, Norma; Olivares-Cervantes, Alma L; Salinas, Eva; Martínez, Leticia; Escorcia, Magdalena; Oropeza, Ricardo; Rosas, Irma
Coagulase-positive staphylococci (CoPS) are opportunistic pathogens carrying various mechanisms of resistance that have a large number of virulence factors, and whose ability to induce illness is associated with the host. This study aimed to investigate the presence of environmental coagulase-positive staphylococci, their susceptibility profile, clonal relationship and ability to form biofilm. The 16S rRNA genes from CoPS isolates were analyzed, and their antibiotic susceptibility was evaluated using the agar dilution method in accordance with Clinical and Laboratory Standards Institute guidelines. The clonal profile was obtained by pulsed-field gel electrophoresis (PFGE) and biofilm formation was measured by a crystal violet retention assay. A total of 72 Staphylococcus spp. strains were isolated from air, metal surfaces, and nostrils from humans, dogs, cats, and birds. Three species were identified: Staphylococcus aureus (17%), Staphylococcus intermedius (63%), and Staphylococcus pseudintermedius (21%). Ninety three percent (93%) of the strains were resistant to at least one of 13 tested antibiotics. S. pseudintermedius strains were the only resistant ones to methicillin while most of these isolates were multidrug-resistant, had significantly higher ability to form biofilm and PFGE grouped into seven different patterns, without showing clonal dispersion among animals and environmental isolates. This study suggests that dogs, cat, and air are environmental sources potentially carrying multidrug-resistant S. pseudintermedius, which survives in different environments through biofilm formation and multidrug resistance, characteristics that can be transmitted horizontally to other bacteria and exacerbate the problem of antibiotic resistance in humans. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.
Lerbeck, Anne Mette; Tersbøl, Britt Pinkowski; Styrishave, Bjarne
596 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......Development of antimicrobial resistance has been assigned to excess and misuse of antimicrobial agents. Staphylococci are part of the normal flora but are also potential pathogens that have become essentially resistant to many known antibiotics. Resistances in coagulase negative staphylococci (Co...
Lerbech, Anne Mette; Opintan, Japheth A; Bekoe, Samuel Oppong
= 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......Development of antimicrobial resistance has been assigned to excess and misuse of antimicrobial agents. Staphylococci are part of the normal flora but are also potential pathogens that have become essentially resistant to many known antibiotics. Resistances in coagulase negative staphylococci (Co...
Full Text Available Introduction. It is very important to know the resistant bacteria to common used antibiotics in our community. Staphylococcus coagulase positive was the main cause of infection in infectious disease. This study was done to demonstrate the pattern of resistance to batalactamase antibiotics among staphiococci. Methods. During a period of five month, 38 coagulase positive staphylococcus isolates were identified from various clinical specimens from 600 patients at the AI-Zahra university hospital (affiliated to IUMSHS. Results. Betalactamase production assays using rapid acidometric and iodometric tests showed that 78.9 percent and 73.6 percent of isolates were positive, respectively. The differnce in determination rate between acidometric and iodometric tests was not statistically significant. Moreover the acidometric test was cheaper and more easy to perform than iodometric test. In vitro sensitivity testing using the disc diffusion method showed that all of isolates were resistant to carbenicillin, ampicillin and amoxicillin, while 7.9 percent and 13.2 percent were resistant to cefazolin and cephalexin, respectively. Discussion. We recommend use of cefazolin, cephalexin and oxacillin for treatment of patients with staphylococcus infections.
Amin, V. M.; Olson, N. F.
Catalase activities of intact cells and cell-free extracts of coagulase-positive staphylococcal cultures 105B and 558D isolated from milk, culture 25042 from a clinical source, and Staphylococcus aureus 196E were determined at 32.2 C. Cultures were treated with 0.025 and 0.05% hydrogen peroxide at 37.8 and 54.4 C and without hydrogen peroxide at 54.4 C to determine the relationship between catalase activity and resistance to these treatments. The relationship held true for cultures 105B and 196E; culture 105B had the lowest catalase activity and lowest resistance to H2O2 at 37.8 C, whereas S. aureus 196E possessed a high catalase activity and was most resistant at 37.8 C. Catalase activities of cell-free extracts of cultures 25042, 558, and 196E were similar, but resistance to H2O2 at 37.8 C was greater for culture 196E. The lower resistance of culture 25042 was related to low catalase activities of whole cells of this culture, which were only one-third that of whole cells of culture 196E. Culture 558 was least resistant to heat treatment at 54.4 C and showed the greatest sensitivity to added H2O2 at this temperature. PMID:5645413
Walther Frans J
Full Text Available Abstract Background Coagulase negative staphylococci (CoNS are the most common cause of neonatal sepsis in the Neonatal Intensive Care Unit (NICU. A minority of neonates does not respond to vancomycin therapy and develops persistent bacteremia, which may be treated with rifampin. We evaluated the use of rifampin in persistent CoNS bacteremia. Methods Retrospective study of 137 neonates with CoNS bacteremia during admission to a tertiary NICU between July 2006 and July 2009. Main outcome measures were total duration of bacteremia and the adequacy of vancomycin and rifampin therapy. Results 137/1696 (8.0% neonates developed a CoNS bacteremia. Eighteen were treated with rifampin because of persistent bacteremia (3 positive blood cultures at least 48 hours apart with clinical symptoms or (a serious suspicion of an intravascular thrombus. Duration of bacteremia prior to rifampin therapy (8.0 ± 3.6 days was positively correlated (p Conclusion Rifampin may be effective in the treatment of persistent CoNS infections in neonates. Outcome may be improved by adequate monitoring of vancomycin trough levels.
Rota, Ada; Corrò, Michela; Drigo, Ilenia; Bortolami, Alessio; Börjesson, Stefan
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
The nasal carriage and antibiotic susceptibility pattern of coagulase negative staphylococci (CoNS) isolates from healthy pigs and in-contact humans were investigated in 300 pigs and 101 in-contact humans from pig farms and abattoir. The results indicate that 13.2% (53/401) of the isolates were CoNS species based on ...
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 ...
Baĭl'ozov, D; Georgiev, L; Zakhariev, Ts
Studied were 382 coagulase-positive and 290 coagullase-negative Staphylococcus strains isolated from birds during their post-slaughter handling, from the vats for water cooling as well as from the women working on the slaughter-house conveyor. Studied were the phage behaviour, fibrinolysine production, the crystal-violet test and the type of hemolysines. It was established that the coagulase positive staphylococci were lysed slightly by the human and the bovine set of phages. The main sources of staphylococci of both human and bovine origin appeared to be the nasal cavity and the hands of workers. Evisceration proved to contribute to the tenfold higher count of staphylococci in the slaughtered birds. Most of the investigated strains of Staphilococcus aureus and Staphylococcus epidermidis differed from the human and the bovine type and were probably staphylococci of avian origin.
Klapes, N. Arlene; Vesley, Donald
A rapid, in situ thermonuclease test that identifies colonies of Staphylococcus aureus among staphylococci isolated from swimming pool water by membrane filtration recovery on various selective and differential media is described.
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
The use of antibiotics in farm management (growing crops and raising animals) has become a major area of concern. Its implications is the consequent emergence of antibiotic resistant bacteria (ARB) and accordingly their access into the human food chain with passage of antibiotic resistance genes (ARG) to the normal human intestinal microbiota and hence to other pathogenic bacteria causative human disease. Therefore, we pursued in this study to unravel the frequency and the quinolone resistance determining region, mecA and cfr genes of methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA), methicillin-resistant coagulase-negative staphylococci (MRCNS) and methicillin-susceptible coagulase-negative staphylococci (MSCNS) isolated from the retail trade of ready-to-eat raw chicken meat samples collected during 1 year and sold across the Great Cairo area. The 50 Staphylococcus isolated from retail raw chicken meat were analyzed for their antibiotic resistance phenotypic profile on 12 antibiotics (penicillin, oxacillin, methicillin, ampicillin-sulbactam, erythromycin, tetracycline, clindamycin, gentamicin, ciprofloxacin, chloramphenicol, sulfamethoxazole-trimethoprim, and vancomycin) and their endorsement of the quinolone resistance determining region, mecA and cfr genes. The isolation results revealed 50 isolates, CPS (14) and CNS (36), representing ten species (S. aureus, S. hyicus, S. epidermedius, S. lugdunensis, S. haemolyticus, S. hominus, S. schleiferi, S. cohnii, S. intermedius, and S. lentus). Twenty seven isolates were methicillin-resistant. Out of the characterized 50 staphylococcal isolates, three were MRSA but only 2/3 carried the mecA gene. The ARG that bestows resistance to quinolones, β-lactams, macrolides, lincosamides, and streptogramin B [MLS(B)] in MRSA and MR-CNS were perceived. According to the available literature, the present investigation was a unique endeavor into the identification of the quinolone
Kamelia Mahmoud Osman
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
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
The use of antibiotics in farm management (growing crops and raising animals) has become a major area of concern. Its implications is the consequent emergence of antibiotic resistant bacteria (ARB) and accordingly their access into the human food chain with passage of antibiotic resistance genes (ARG) to the normal human intestinal microbiota and hence to other pathogenic bacteria causative human disease. Therefore, we pursued in this study to unravel the frequency and the quinolone resistance determining region, mecA and cfr genes of methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA), methicillin-resistant coagulase-negative staphylococci (MRCNS) and methicillin-susceptible coagulase-negative staphylococci (MSCNS) isolated from the retail trade of ready-to-eat raw chicken meat samples collected during 1 year and sold across the Great Cairo area. The 50 Staphylococcus isolated from retail raw chicken meat were analyzed for their antibiotic resistance phenotypic profile on 12 antibiotics (penicillin, oxacillin, methicillin, ampicillin-sulbactam, erythromycin, tetracycline, clindamycin, gentamicin, ciprofloxacin, chloramphenicol, sulfamethoxazole-trimethoprim, and vancomycin) and their endorsement of the quinolone resistance determining region, mecA and cfr genes. The isolation results revealed 50 isolates, CPS (14) and CNS (36), representing ten species (S. aureus, S. hyicus, S. epidermedius, S. lugdunensis, S. haemolyticus, S. hominus, S. schleiferi, S. cohnii, S. intermedius, and S. lentus). Twenty seven isolates were methicillin-resistant. Out of the characterized 50 staphylococcal isolates, three were MRSA but only 2/3 carried the mecA gene. The ARG that bestows resistance to quinolones, β-lactams, macrolides, lincosamides, and streptogramin B [MLS(B)] in MRSA and MR-CNS were perceived. According to the available literature, the present investigation was a unique endeavor into the identification of the quinolone
Lopez, Julien; Tatar, Zuzana; Tournadre, Anne; Couderc, Marion; Pereira, Bruno; Soubrier, Martin; Dubost, Jean-Jacques
Coagulase-negative staphylococci (CoNS) are increasingly implicated in recent patient series of spondylodiscitis, but there are no series of CoNS-spondylodiscitis available. The objective of this study was to compare the characteristics of patients with spontaneous CoNS-spondylodiscitis with those patients with Staphylococcus aureus (SA) spondylodiscitis. This was a retrospective single center study involving 147 spontaneous infectious spondylodiscitis cases observed between 2000 and 2015. The 26 cases of CoNS-spondylodiscitis (15 confirmed) were compared with 30 cases of SA-spondylodiscitis. CoNS infection was considered confirmed if the same CoNS was isolated in at least two samples at two different times. Patients with CoNS-spondylodiscitis were older (70 vs. 61 years of age; p = 0.01), had associated cancer more often (15% vs. 0%; p = 0.04) and had a longer diagnostic delay (>15 days in 88% vs. 60%; p = 0.01); experienced fever less often (19% vs. 50%; p = 0.01), and had lower white blood cell (7.6 vs. 9.9G/L; p = 0.01) and polymorphonuclear leucocyte counts (5.6 vs. 7.5G/L; p = 0.04). Patients with CoNS spondylodiscitis had less pronounced inflammatory syndrome (erythrocyte sedimentation rate [ESR]: 62 vs. 81 mm at 1 h; p = 0.03; CRP: 60 vs. 147 mg/L; p = 0.0003) and less common (ESR < 30 mm: 23% vs. 0%; p = 0.01; CRP < 10 mg/L: 23% vs. 0%; p = 0.005) in comparison with patients with SA infection. The infection entry site was most often an intravascular catheter (20% vs. 3%; p = 0.008). The level of positive percutaneous needle biopsies was comparable between CoNS and SA. Two patients who died both had SA infections. CoNS-spondylodiscitis involved at least 10% of spontaneous spondylodiscitis cases and was more common in elderly patients, afflicted by comorbidities, and its presentation was less virulent than that of those with SA-spondylodiscitis.
Rasmussen, Rasmus V; Snygg-Martin, Ulrika; Olaison, Lars
The aim of this study was to investigate in-hospital mortality and 12-month mortality in patients with coagulase-negative Staphylococcus (CoNS) compared to Staphylococcus aureus (S. aureus) infective endocarditis (IE). We used a prospective cohort study of 66 consecutive CoNS and 170 S. aureus IE...
ESQUEMA SIMPLIFICADO PARA IDENTIFICAÇÃO DE ESTAFILOCOCOS COAGULASE-POSITIVOS ISOLADOS DE MASTITE BOVINA SIMPLIFIED SCHEME FOR IDENTIFICATION OF COAGULASE-POSITIVE STAPHYLOCOCCI ISOLATED FROM BOVINE MASTITIS
Maria Aparecida Vasconcelos Paiva Brito
to bacitracin, aerobic acid from glycerol in the presence of 0.4mug m-1 of eritromicin, catalase, and coagulated rabbit plasma by the tube test. The acriflavin susceptibility was tested on Baird Parker and P agar plates containing 7mug m-1 of acriflavin. Five coagulase-negative S. hyicus strains identified by the API Staph system and S. aureus ATCC 29213 were used as test controls. Thirty-eight out of the 49 CPS were identified as S. aureus and 11 as S. hyicus, while S. intermedius was not found. The API Staph system was used to confirm the identification of the coagulase-positive strains of S. hyicus, seven S. aureus strains not producing acetoin and four negative for anaerobic fermentation of mannitol. All the strains identified as S. aureus were resistant to acriflavin whereas S. hyicus were sensitive. It was concluded that the sensitivity to acriflavin can be used in addition to the tests of coagulase and acetoin production to differentiate CPS isolated from bovine mastitis.
Full Text Available Clindamycin has been used successfully to treat pneumonia and soft-tissue infections caused by methicillin-resistant Staphylococcus aureus. However, inducible clindamycin resistance has been described as a cause of treatment failure of such infections. A total of 159 staphylococcal isolates from different clinical specimens from burn patients in Tripoli Burn Center were tested for inducible clindamycin resistance by the disk-diffusion induction test. Inducible clindamycin resistance was detected in 66.2% of 65 methicillin-resistant S. aureus isolates and in none of 55 methicillin-sensitiveS. aureus, 10 methicillin-resistant coagulase negative staphylococci and 29 methicllin-sensitive coagulase negative staphylococci isolates. In our setting, clindamycin can be used for the treatmentof infections due to staphylococci, but we recommend that staphylococci isolates, particularly methicillin-resistant S. aureus, are tested by the D-test before treatment.
Lenart-Boroń, Anna; Wolny-Koładka, Katarzyna; Stec, Joanna; Kasprowic, Andrzej
This study assessed the antimicrobial resistance of airborne Staphylococcus spp. strains isolated from healthcare facilities in southern Poland. A total of 55 isolates, belonging to 10 coagulase-negative staphylococci (CoNS) species, isolated from 10 healthcare facilities (including hospitals and outpatient units) were included in the analysis. The most frequently identified species were Staphylococcus saprophyticus and Staphylococcus warneri, which belong to normal human skin flora, but can also be the cause of common and even severe nosocomial infections. Disk diffusion tests showed that the bacterial strains were most frequently resistant to erythromycin and tetracycline and only 18% of strains were susceptible to all tested antimicrobials. Polymerase chain reaction amplification of specific gene regions was used to determine the presence of the Macrolide-Lincosamide-Streptogramin resistance mechanisms in CoNS. The molecular analysis, conducted using specific primer pairs, identified the msrA1 gene, encoding active efflux pumps in bacterial cells, as the most frequent resistance gene. As many as seven antibiotic resistance genes were found in one isolate, whereas the most common number of resistance genes per isolate was five (n = 17). It may be concluded that drug resistance was widely spread among the tested strains, but the resulting antimicrobial resistance profile indicates that in the case of infection, the use of antibiotics from the basic antibiogram group will be effective in therapy. However, before administering treatment, determination of the specific antimicrobial resistance should be conducted, particularly in the case of hospitalized patients.
Beenen, L F; Touw, D J; Hekker, T A; Haring, D A
Following craniotomy for a medulloblastoma in the posterior cranial fossa, a 6-year old girl developed a ventriculitis with coagulase negative staphylococci associated with the use of a ventriculostomy. Treatment with intravenous (i.v.) and intraventricular (ivt) vancomycin resulted in negative
Beenen, L. F.; Touw, D. J.; Hekker, T. A.; Haring, D. A.
Following craniotomy for a medulloblastoma in the posterior cranial fossa, a 6-year old girl developed a ventriculitis with coagulase negative staphylococci associated with the use of a ventriculostomy. Treatment with intravenous (i.v.) and intraventricular (ivt) vancomycin resulted in negative
Maria de Fátima Azevedo Gayoso
Full Text Available OBJETIVO: Avaliar a suscetibilidade, in vitro, de Staphylococcus coagulase negativa (SCoN, isolados da conjuntiva e córnea, à meticilina, às fluoroquinolonas e aos aminoglicosídeos. MÉTODOS: Foram analisadas retrospectivamente 707 amostras oculares de SCoN quanto à suscetibilidade aos antimicrobianos pelo teste de disco difusão, durante o período de janeiro de 2000 a dezembro de 2003. RESULTADOS: Houve um aumento do número de SCoN em isolados da conjuntiva (n=57, ano de 2000 e n=153, ano de 2003 e da córnea (n=28, ano de 2000 e n=78, ano de 2003. A freqüência de SCoN resistentes à meticilina isolados da conjuntiva e da córnea, aumentou (1,8 a 19,6% e 14,3 a 29,3% respectivamente durante o período avaliado. Não houve diferença estatisticamente significante nos anos estudados, nos percentuais de SCoN resistentes às fluoroquinolonas, nas conjuntivas (ofloxacina: 1,8 a 7,8% e ciprofloxacina: 3,5 a 9,2% e nas córneas (ofloxacina: 14,3 a 9,0% e ciprofloxacina:14,3 a 10,3%. Avaliando-se os resultados das amostras isoladas da conjuntiva, observou-se um aumento na resistência à tobramicina: 15,8 a 34,6%; e à gentamicina: 10,5 a 25,5%; mas não houve mudança no perfil de resistência das amostras da córnea à tobramicina: 28,6 a 26,9% e à gentamicina: 21,4 a 23,1%. CONCLUSÃO: Houve diminuição na suscetibilidade in vitro dos SCoN para meticilina, tobramicina e gentamicina. As fluoroquinolonas, representadas pela ofloxacina e ciprofloxacina, demonstraram ter um padrão estável de suscetibilidade in vitro.PURPOSE: To assess the in vitro susceptibility of conjunctival and corneal coagulase negative Staphylococcus (CoNS to methicillin, fluoroquinolones and aminoglycosides. METHODS: A total of 707 conjunctival and corneal CoNS disk diffusion test results were retrospectively analyzed, from January 2000 through December 2003. RESULTS: From 2000 to 2003, there was an increase in number of CoNS isolated from conjunctiva (n=57 to n=153
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 ...
Terpstra, S; Noordhoek, GT; Voesten, HGJ; Degener, JE
One approach for prosthetic vascular surgery is to continue antimicrobial prophylaxis while intravascular lines and catheters are in place. However this may give rise to antimicrobial resistance in the colonizing bacterial flora. We studied 37 patients undergoing vascular surgery, who received
Soge, Olusegun O; Meschke, John S; No, David B; Roberts, Marilyn C
The aim of this study was to isolate and characterize methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative Staphylococcus spp. (MRCoNS) from marine water and intertidal beach sand from public beaches in Washington State, USA. Fifty-one staphylococci from Washington State beaches were characterized using antimicrobial susceptibility testing, carriage of acquired tetracycline and/or macrolide resistance genes, staphylococcal cassette chromosome mec (SCCmec) typing, the BBL Crystal Gram-Positive ID System and/or 16S rRNA sequencing, coagulase test and multilocus sequence typing (MLST) for MRSA. Five multidrug-resistant MRSA SCCmec type I, of which three were MLST type ST45, one ST59 and one a new MLST type, ST1405, plus one susceptible non-typeable (NT) MRSA ST30 were characterized. Thirty-three MRCoNS isolates, representing 21 strains from 9 Staphylococcus spp., carried a range of SCCmec types [I (2), II (6), III (3), V (2), I/II (1) and NT (7)] and varied in their antibiotic susceptibility to other antibiotic classes and carriage of acquired tetracycline/macrolide resistance gene(s). MRSA and MRCoNS donors co-transferred tet(M) and erm(A) genes to an Enterococcus faecalis recipient at a frequency of 10(-8). This is the first report of MRSA and MRCoNS isolated from marine water and intertidal beach sand. The MLST types and antibiotic carriage of five MRSA isolates were similar to hospital MRSA isolates rather than US community-acquired MRSA isolates. Our results suggest that public marine beaches may be a reservoir for transmission of MRSA to beach visitors as well as an ecosystem for exchange of antibiotic resistance genes among staphylococci and related genera.
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
Walther, Birgit; Hermes, Julia; Cuny, Christiane; Wieler, Lothar H; Vincze, Szilvia; Abou Elnaga, Yassmin; Stamm, Ivonne; Kopp, Peter A; Kohn, Barbara; Witte, Wolfgang; Jansen, Andreas; Conraths, Franz J; Semmler, Torsten; Eckmanns, Tim; Lübke-Becker, Antina
Since the relationship between dogs and their owners has changed, and dogs moved from being working dogs to family members in post-industrial countries, we hypothesized that zoonotic transmission of opportunistic pathogens like coagulase positive staphylococci (CPS) is likely between dogs and their owners. CPS- nasal carriage, different aspects of human-to-dog relationship as well as potential interspecies transmission risk factors were investigated by offering nasal swabs and a questionnaire to dog owners (108) and their dogs (108) at a dog show in 2009. S. aureus was found in swabs of 20 (18.5%) humans and two dogs (1.8%), and spa types which correspond to well known human S. aureus lineages dominated (e.g. CC45, CC30 and CC22). Multilocus sequence typing (MLST) of the two canine strains revealed ST72 and ST2065 (single locus variant of ST34). Fifteen dogs (13.9%) and six owners (5.6%) harboured S. pseudintermedius, including one mecA-positive human isolate (MRSP). Pulsed field gel electrophoresis (PFGE) revealed that one dog/owner pair harboured indistinguishable S. pseudintermedius- isolates of ST33. Ten (48%) of the 21 S. pseudintermedius-isolates showed resistance towards more than one antimicrobial class. 88.9% of the dog owners reported to allow at least one dog into the house, 68.5% allow the dog(s) to rest on the sofa, 39.8% allow their dogs to come onto the bed, 93.5% let them lick their hands and 52.8% let them lick their face. Bivariate analysis of putative risk factors revealed that dog owners who keep more than two dogs have a significantly higher chance of being colonized with S. pseudintermedius than those who keep 1-2 dogs (powners and their dogs is possible. Further investigation regarding interspecies transmission and the diverse adaptive pathways influencing the epidemiology of CPS (including MRSA and MRSP) in different hosts is needed.
Differentiation between Staphylococcus aureus and coagulase-negative Staphylococcus species by real-time PCR including detection of methicillin resistants in comparison to conventional microbiology testing.
Klaschik, Sven; Lehmann, Lutz E; Steinhagen, Folkert; Book, Malte; Molitor, Ernst; Hoeft, Andreas; Stueber, Frank
Staphylococcus aureus has long been recognized as a major pathogen. Methicillin-resistant strains of S. aureus (MRSA) and methicillin-resistant strains of S. epidermidis (MRSE) are among the most prevalent multiresistant pathogens worldwide, frequently causing nosocomial and community-acquired infections. In the present pilot study, we tested a polymerase chain reaction (PCR) method to quickly differentiate Staphylococci and identify the mecA gene in a clinical setting. Compared to the conventional microbiology testing the real-time PCR assay had a higher detection rate for both S. aureus and coagulase-negative Staphylococci (CoNS; 55 vs. 32 for S. aureus and 63 vs. 24 for CoNS). Hands-on time preparing DNA, carrying out the PCR, and evaluating results was less than 5 h. The assay is largely automated, easy to adapt, and has been shown to be rapid and reliable. Fast detection and differentiation of S. aureus, CoNS, and the mecA gene by means of this real-time PCR protocol may help expedite therapeutic decision-making and enable earlier adequate antibiotic treatment. © 2014 Wiley Periodicals, Inc.
Boo, N Y; Suhaida, A R; Rohana, J
This case-control study aimed to determine whether catheter use was significantly associated with coagulase-negative staphylococci (CoNS) colonisation and/or sepsis in neonates. Weekly swabs of the nose, umbilicus, rectum, wounds, eye discharge and intravenous catheter tips (after removal) of infants admitted to the neonatal intensive care unit of Universiti Kebangsaan Malaysia Medical Centre, Malaysia, were cultured. CoNS sepsis was diagnosed if pure growth of CoNS was cultured from the peripheral blood specimen of symptomatic infants. For each infant with CoNS colonisation or sepsis, a control infant was retrospectively and randomly selected from unaffected infants in the ward. Multivariate analyses were performed to determine whether catheter use was a significant risk factor. CoNS colonisation was detected in 113 (8.7%) infants. CoNS sepsis was found in 12 (10.6%) infants with CoNS colonisation and 7 (0.6%) infants without CoNS colonisation. Multivariate analysis showed that the following were significantly associated with CoNS colonisation: conjunctivitis (adjusted odds ratio [OR] 8.2, 95% confidence interval [CI] 1.9–34.8, p = 0.005); central venous catheters (adjusted OR 5.8, 95% CI 1.9–17.8, p = 0.002); and nasopharyngeal and/or oral suctioning more than twice in the 48 hours before positive culture (adjusted OR 7.3, 95% CI 3.3–16.2, p colonisation and sepsis.
Park, Joo Youn; Fox, Lawrence K; Seo, Keun Seok; McGuire, Mark A; Park, Yong Ho; Rurangirwa, Fred R; Sischo, William M; Bohach, Gregory A
Coagulase-negative staphylococci (CNS) are the most frequently isolated pathogens from cows with intramammary infection (IMI). Although API STAPH ID 20, a commercially available identification system, and PCR-restriction fragment length polymorphism (PCR-RFLP) of the gap gene (gap PCR-RFLP) have been successfully applied for the identification of CNS isolates from human specimens, their accuracy in the identification of veterinary isolates has not been fully established. In this study, we identified 263 CNS isolates from bovine IMI at species level by partial 16S rRNA gene sequence analysis as the definitive test. Species identification obtained using partial 16S rRNA gene sequence analysis was compared to results from the API STAPH ID 20 and gap PCR-RFLP analysis. Eleven different CNS species were identified by partial 16S rRNA gene sequence analysis. Only 76.0% (200/263) of the species identification results obtained by API STAPH ID 20 matched those obtained by partial 16S rRNA gene sequence analysis, whereas 97.0% (255/263) of the species identification results obtained by the gap PCR-RFLP analysis matched those obtained by partial 16S rRNA gene sequence analysis. The gap PCR-RFLP analysis could be a useful and reliable alternative method for the species identification of CNS isolates from bovine IMI and appears to be a more accurate method of species identification than the API STAPH ID 20 system. Copyright © 2010 Elsevier B.V. All rights reserved.
De Visscher, A; Piepers, S; Haesebrouck, F; De Vliegher, S
Coagulase-negative staphylococci (CNS) are the main cause of bovine intramammary infections and are also abundantly present in extramammary habitats such as teat apices. Teat apex colonization (TAC) with CNS has already been explored in lactating dairy cows at the species level, whereas this is not true for dry cows and end-term heifers. Therefore, the aim of this observational study was to describe CNS TAC in nonlactating dairy cows and end-term heifers in Flemish dairy herds and to identify associated risk factors at the herd, cow, and quarter level. All CNS were molecularly identified to the species level using transfer RNA intergenic spacer PCR (tDNA-PCR) and sequencing of the 16S rRNA gene, allowing for species-specific statistical analyses using multivariable, multilevel logistic regression. Staphylococcus devriesei, Staphylococcus chromogenes, Staphylococcus haemolyticus, and Staphylococcus equorum were the most frequently isolated species. Staphylococcus chromogenes was the sole species colonizing teat apices of cows and heifers in all herds, whereas large between-herd differences were observed for the other species. Teat apices of red and white Holstein Friesians, of quarters dried off without an internal teat sealer, and swabbed in months with lower precipitation and higher ambient temperature were significantly more likely to be colonized by S. devriesei. Slightly dirty teat apices and teat apices swabbed in months with lower precipitation had higher odds of being colonized by S. chromogenes, whereas teat apices sampled in months with lower precipitation and higher ambient temperature were more likely to be colonized by S. haemolyticus. Dirty teat apices and teat apices swabbed in months with lower ambient temperature in combination with low precipitation had higher odds of being colonized by S. equorum. Diverse factors explaining CNS TAC, yet mostly related to humidity, ambient temperature, and hygiene, substantiate differences in epidemiological
Moura,Tiane Martin de; Campos,Fabrício Souza; d'Azevedo,Pedro Alves; Van Der Sand,Sueli Teresinha; Franco,Ana Cláudia; Frazzon,Jeverson; Frazzon,Ana Paula Guedes
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 staphyl...
Full Text Available BACKGROUND: Since the relationship between dogs and their owners has changed, and dogs moved from being working dogs to family members in post-industrial countries, we hypothesized that zoonotic transmission of opportunistic pathogens like coagulase positive staphylococci (CPS is likely between dogs and their owners. METHODOLOGY/PRINCIPAL FINDINGS: CPS- nasal carriage, different aspects of human-to-dog relationship as well as potential interspecies transmission risk factors were investigated by offering nasal swabs and a questionnaire to dog owners (108 and their dogs (108 at a dog show in 2009. S. aureus was found in swabs of 20 (18.5% humans and two dogs (1.8%, and spa types which correspond to well known human S. aureus lineages dominated (e.g. CC45, CC30 and CC22. Multilocus sequence typing (MLST of the two canine strains revealed ST72 and ST2065 (single locus variant of ST34. Fifteen dogs (13.9% and six owners (5.6% harboured S. pseudintermedius, including one mecA-positive human isolate (MRSP. Pulsed field gel electrophoresis (PFGE revealed that one dog/owner pair harboured indistinguishable S. pseudintermedius- isolates of ST33. Ten (48% of the 21 S. pseudintermedius-isolates showed resistance towards more than one antimicrobial class. 88.9% of the dog owners reported to allow at least one dog into the house, 68.5% allow the dog(s to rest on the sofa, 39.8% allow their dogs to come onto the bed, 93.5% let them lick their hands and 52.8% let them lick their face. Bivariate analysis of putative risk factors revealed that dog owners who keep more than two dogs have a significantly higher chance of being colonized with S. pseudintermedius than those who keep 1-2 dogs (p<0.05. CONCLUSIONS/RECOMMENDATIONS: In conclusion, CPS transmission between dog owners and their dogs is possible. Further investigation regarding interspecies transmission and the diverse adaptive pathways influencing the epidemiology of CPS (including MRSA and MRSP in
N Bhaskar Prabhu
Full Text Available A total of 109 cases of pyoderma were studied bacteriologicaliy by culture. Folliculitis cox~stituted the largest clinical group followed by impetigo and eczematoid dermatitis. Coagulase positive staphylococci were found to be the predominant aetiological agents in pure form and also in association with other infecting agents. Coagulase negative staphylococci constituted the next common organism, followed by beta-haemolytic streptococci. Nasal and throat swabs from all the cases were subjected to bacteriologic study and the association of nasal and throat staphylococci with pyoderma lesions was studied. Majority (56.66% of staph-aureus strains were non typable; 56.66% of typable strains belonged to phage type III, 26.67% to phage type II and 16.67% to phage type I. The throat staphylococcal phage types were found to be associated more with pyoderma than nasal phage types.
Turutoglu H.; Mudul S.; Pehlivanoglu F.
In this study, it was aimed to investigate the mastitis pathogens and to determine the b-lactamase activities of staphylococci isolated from bovine mastitis milk samples and the sensitivities of these isolates to various antibiotics. For this purpose, 1180 dairy cows On 131 farms were examined for mastitis and 496 mastitis milk samples were taken 'from 249 cows. Staphylococcus aureus was present in 33.16%, coagulase negative staphylococci in 31.10 %, Eschehchia coli in 11.57 %, S...
T V Ramani
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.
De Buyser, M L; Lombard, B; Schulten, S M; In't Veld, P H; Scotter, S L; Rollier, P; Lahellec, C
The methods of European and International Organisations for Standardization for the enumeration of coagulase-positive staphylococci (CPS, Staphylococcus aureus and other species) described in EN ISO 6888 Part 1 and Part 2: 1999 were validated by order of the European Commission (Standards, Measurement and Testing Fourth Framework Programme Project SMT4-CT96-2098). EN ISO 6888-1 prescribes the use of Baird-Parker (BP) agar whereas EN ISO 6888-2 prescribes the use of Rabbit Plasma Fibrinogen Agar (RPFA). The objective was to determine the precision of each method in terms of repeatability (r) and reproducibility (R) using three different food types inoculated with various levels of S. aureus and a typical background flora. The results are intended for publication in the associated standards. Cheese, meat and dried egg powder were examined by 24 laboratories from 16 countries in Europe. Each participant received eight test materials per food type: blind duplicates at four inoculum levels (0, 10(3), 10(4) to 10(5), 10(5) to 10(6) cfu/g). In addition, two reference materials (RM) (capsules containing milk powder inoculated with S. aureus) were included in the study. All test materials were subjected to stringent homogeneity and stability testing before being used in the collaborative trial. Two statistical methods were used to calculate the precision parameters. Draft EN ISO 16140: 2000 method appeared more appropriate to the case of microbiological data than ISO 5725-2: 1994 method and was retained to calculate the precision data. Concerning EN ISO 6888-1, overall values for repeatability (r) when used with food test materials was r=log(10) 0.28 (expressed as an absolute difference between log(10)-transformed test results). For the reference materials, r=log(10) 0.19. Overall values for reproducibility (R) when used with food test materials were R=log(10) 0.43. For the reference materials, R=log(10) 0.39. Concerning EN ISO 6888-2, overall values for repeatability (r
Flamm, Robert K; Rhomberg, Paul R; Kaplan, Nachum; Jones, Ronald N; Farrell, David J
Staphylococcus aureus and coagulase-negative staphylococci (CoNS) are responsible for a wide variety of human infections. The investigational antibacterial Debio1450 (previously AFN-1720), a prodrug of Debio1452 (previously AFN-1252), specifically targets staphylococci without significant activity against other Gram-positive or Gram-negative species. Debio1452 inhibits FabI, an enzyme critical to fatty acid biosynthesis in staphylococci. The activity of Debio1452 against CoNS, methicillin-susceptible S. aureus (MSSA), and methicillin-resistant S. aureus (MRSA), including significant clones, was determined. A globally diverse collection of 574 patient isolates from 35 countries was tested that included CoNS (6 species, 103 strains), MSSA (154 strains), MRSA (163 strains), and molecularly characterized strains (including spa-typed MRSA clones; 154 strains). The isolates were tested for susceptibility by CLSI broth microdilution methods against Debio1452 and 10 comparators. The susceptibility rates for the comparators were determined using CLSI and EUCAST breakpoint criteria. All S. aureus and CoNS strains were inhibited by Debio1452 concentrations of ≤ 0.12 and ≤ 0.5 μg/ml, respectively. The MIC50s for MSSA, MRSA, and molecularly characterized MRSA strains were 0.004 μg/ml, and the MIC90s ranged from 0.008 to 0.03 μg/ml. The MICs were higher for the CoNS isolates (MIC50/90, 0.015/0.12 μg/ml). Among S. aureus strains, resistance was common for erythromycin (61.6%), levofloxacin (49.0%), clindamycin (27.6%), tetracycline (15.7%), and trimethoprim-sulfamethoxazole (7.0%). Debio1452 demonstrated potent activity against MSSA, MRSA, and CoNS. Debio1452 showed significantly greater activity overall (MIC50, 0.004 μg/ml) than the other agents tested against these staphylococcal species, which included dominant MRSA clones and strains resistant to currently utilized antimicrobial agents. Copyright © 2015, American Society for Microbiology. All Rights Reserved.
Cybulska, Janina; Jeljaszewicz, J.
Cybulska, Janina (State Institute of Hygiene, Warsaw, Poland), and J. Jeljaszewicz. Bacteriostatic activity of serum against staphylococci. J. Bacteriol. 91:953–962. 1966.—Antistaphylococcal activity of normal serum against strains exhibiting various patterns of coagulase, clumping-factor, and staphylokinase production is not connected with the presence of these factors. Purified coagulase does not influence this property of serum. Coagulase-negative strains with clumping-factor activity grow in normal serum as typical pathogenic staphylococci. Serum bacteriostatic activity against staphylococci may be reversed by several nonspecific factors, such as sterile broth, supernatant fluids of coagulase-negative strains, and ammonium sulfate precipitates of culture supernatant fluids of various staphylococci. Immune sera with a high agglutinating titer for staphylococcal cells do not prevent growth of serum-resistant strains; serum-susceptible strains are inhibited as in normal serum control. Activation or blocking of the serum fibrinolytic system does not influence serum bacteriostatic activity. The growth rate of serum-resistant strains is identical in serum and in Todd-Hewitt broth; serum-susceptible strains are inhibited to the inoculum level, but decreases and increases in viable count are noted during a 24-hr observation period. Observations made with sera of 10 animal species clearly demonstrated differences in serum bacteriostatic activity, mouse serum being completely noninhibitory and cat serum only weakly inhibitory. The technique of quantitative determination of serum susceptibility of staphylococci is described, and the importance of serum antistaphylococcal activity in vitro is discussed. Experimental staphylococcal infection produced in rabbits by intravenous injection of different Staphylococcus aureus strains did not result in significant changes in serum antistaphylococcal activity. The technique of experimental infection used caused chronic
E. J. Ridgway; A.P.R. Wilson; M. C. Kelsey
textabstractCultures of nasal or presternal swabs form part of the routine preoperative screening of patients on the cardiac surgical ward. During a trial of antibiotic prophylaxis in 314 patients, preoperative isolates of Staphylococcus aureus and coagulase-negative staphylococci were compared with
Zemelman, Raúl; Longeri, Luis
To evaluate the pathogenicity of staphylococci from bovine raw milk, the general characteristics of 775 strains isolated from 798 samples of milk were studied. The coagulase test was performed by use of rabbit plasma. Chromogenesis, mannitol fermentation, and gelatin liquefaction were investigated on Chapman's Medium 110, after 48 hr of incubation. Production of β-hemolysin, which has been considered indicative of pathogenic staphylococci of animal origin, was determined by streaking different strains on sheep blood-agar plates in the presence of a strain of Lancefield group B streptococci. Plates were incubated at 37 C for 24 hr, and strong hemolysis was produced in the zone of interaction of β-hemolysin and some substance liberated by streptococcus (CAMP test). Of 404 strains found to be coagulase-positive, 95.8% exhibited a deep-orange pigment, 76.5% produced β-hemolysin, 91.8% fermented mannitol, and 75% liquefield gelatin. Of 371 strains which gave a negative coagulase test, about 16% fermented mannitol and liquefied gelatin; none of these strains produced β-hemolysin. When results are grouped according to pigmentation and coagulase production, β-hemolysin seems to be developed by pathogenic strains of Staphylococcus aureus only. If suitability of these tests for investigation of pathogenicity is compared, production of β-hemolysin appears to be the most useful one, since no “false positive” results were found. The use of the CAMP test as a simple and rapid technique to determine production of β-hemolysin by pathogenic strains of animal staphylococci during routine bacteriological work is suggested. Images Fig. 1 PMID:14325873
Davidson, T J; Dohoo, I R; Donald, A W; Hariharan, H; Collins, K
The epidemiology and importance of coagulase negative staphylococcal (CNS) mastitis in Prince Edward Island had not been documented. To investigate this, a cohort of 84 cows at seven farms were quarter sampled eight times over a lactation, commencing with samples taken prior to drying off in the previous lactation. Thirteen species of CNS were isolated. The quarter prevalence of CNS mastitis varied from 4.8% to 6.4% in the first five months of lactation and increased to 14.2 to 16.6% in the last four months of lactation. The geometric mean somatic cell counts (SCC) for quarters infected with CNS and uninfected quarters were 90 x 10(3) and 64 x 10(3) respectively (difference significant at p > 0.005). The two month new infection risk of CNS was 9.0% while the two month elimination risk was 74.4%. Infection with CNS did not alter the risk of subsequent infection with Staphylococcus aureus. The results from this project support the classification of CNS as a minor pathogen in mastitis control programs. PMID:1477796
Ocurrence of Vibrio spp., positive coagulase staphylococci and enteric bacteria in oysters (Crassostrea gigas harvested in the south bay of Santa Catarina island, Brazil Ocorrência de Vibrio spp., estafilococos coagulase positivo e bactérias entéricas em ostras (Crassostrea gigas cultivadas na baía sul da ilha de Santa Catarina, Brasil
Roberta Juliano Ramos
Full Text Available The aim of this study was to assess the contamination of oysters (Crassostrea gigas, harvested in six different regions of the South Bay of Santa Catarina Island, with Coliforms at 45 ºC, Escherichia coli, Vibrio spp., positive coagulase staphylococci, and Salmonella sp. over a period of one year. One hundred eighty oyster samples were collected directly from their culture sites and analyzed. Each sample consisted of a pool of 12 oysters. All of the samples analyzed showed absence of Salmonella, 18 (10% samples showed presence of Escherichia coli, 15 (8.3% samples were positive for V. alginolyticus, and Vibriocholerae was detected in 4 samples (2.2%. The counts of positive-coagulase staphylococci varied from O objetivo deste estudo foi avaliar a contaminação de ostras (Crassostrea gigas cultivadas em diferentes regiões da Baía Sul da Ilha de Santa Catarina, por coliformes a 45 ºC, Escherichia coli, Vibrio spp. Estafilococos coagulase positiva e Salmonella sp., durante o período de um ano. Foram analisadas 180 amostras, coletadas diretamente no local de cultivo. Todas as amostras analisadas apresentaram ausência de Salmonella, 18 (10% amostras apresentaram presença de Escherichia coli, 15 (8,3% amostras positivas para Vibrio alginolyticus e V. cholerae foi detectado em 4 amostras (2,2%. As contagens de Estafilococos coagulase positiva variaram de <10 a 1,9 x 102 UFC.g-1, enquanto que as contagens de coliformes a 45 ºC e E. coli variaram de <3 a 1,5 x 102 NMP.g-1 e <3 e 4,3 x 10 NMP.g-1, respectivamente. As contagens de V. parahaemolyticus e V. vulnificus variaram de <3 a 7 NMP.g-1, para ambos os microrganismos, sugerindo um monitoramento tanto destas espécies quanto da temperatura das águas marinhas nas regiões de cultivo. Com base nos resultados das análises microbiológicas, as amostras analisadas mostraram qualidade bacteriológica aceitável, ou seja, dentro dos parâmetros estabelecidos na legislação brasileira.
Ericson, Jessica E; Thaden, Joshua; Cross, Heather R; Clark, Reese H; Fowler, Vance G; Benjamin, Daniel K; Cohen-Wolkowiez, Michael; Hornik, Christoph P; Smith, P Brian
Coagulase-negative Staphylococcus (CoNS) is the most common cause of bloodstream infections (BSI) in hospitalized infants. CoNS BSI is most reliably treated with vancomycin; however, concerns about side effects and promoting resistance often delay empirical vancomycin therapy until culture results become available. All infants with CoNS BSI discharged from 348 neonatal intensive care units managed by the Pediatrix Medical Group from 1997 to 2012 were identified. Empirical vancomycin therapy was defined as vancomycin exposure on the day of the first positive blood culture. Delayed vancomycin therapy was defined as vancomycin exposure 1-3 days after the first positive blood culture. We used multivariable logistic regression with random effects for site to evaluate the association between the use of empirical vancomycin therapy versus delayed vancomycin therapy and 30-day mortality, controlling for gestational age, small-for-gestational age status, postnatal age on the day of the first positive culture, oxygen requirement, ventilator support and inotropic support on the day the first positive culture was obtained. A total of 4364 infants with CoNS BSI were identified; 2848 (65%) were treated with empirical vancomycin. The median postnatal age at first positive culture was 14 days (interquartile range: 9, 21). Unadjusted 30-day mortality was similar for infants treated with empirical vancomycin and infants treated with delayed vancomycin therapy [166/2848 (6%) vs. 69/1516 (4%); P = 0.08]. There was no significant difference in 30-day mortality on multivariable analysis [odds ratio: 1.14 (0.84, 1.56)]. The median duration of bacteremia was 1 day longer for infants with delayed vancomycin therapy [4 days (interquartile range: 2, 6) vs. 3 days (2, 5); P empirical vancomycin therapy for CoNS BSI was not associated with improved mortality.
Ozkan, Hilal; Cetinkaya, Merih; Koksal, Nilgün; Celebi, Solmaz; Hacımustafaoglu, Mustafa
The aim of this study was to determine the causative agents in early, late- and very late-onset sepsis in preterm infants. The demographic features, risk factors, clinical and laboratory findings in sepsis types were also defined. A total of 151 preterm infants with culture-proven neonatal sepsis were enrolled in this prospective study. The infants were classified into three groups with regard to the onset of sepsis: early onset sepsis (EOS), late-onset sepsis (LOS) and very late-onset sepsis (VLOS). A sepsis screen including whole blood count, blood smear, infection markers and cultures was performed before initiating antibiotic therapy. EOS, LOS and VLOS groups consisted of 23, 86 and 42 infants, respectively. Coagulase-negative staphylococci (CONS) was the most common organism in all sepsis groups. The main factors associated with EOS included presence of premature rupture of membranes, antibiotic use in pregnancy and choriamnionitis. Previous antibiotic use was the main factor associated with LOS, while low birthweight was the main factor in infants with VLOS. Although mortality rate due to Gram-negative bacteria and fungi was higher, CONS was an important cause of mortality in infants with LOS and VLOS. CONS was found to be the most common causative organism in three sepsis types in preterm neonates. Although the mortality rate due to CONS was lower in EOS, it was an important cause of mortality in LOS and VLOS. CONS seems to be the main pathogen in neonatal sepsis in developing countries, as in developed countries. © 2013 The Authors. Pediatrics International © 2013 Japan Pediatric Society.
Blayney, Marc P; Al Madani, Mahmud
Following the introduction of cloxacillin and gentamicin as the first line of treatment for possible late-onset sepsis (LOS) in the authors' neonatal intensive care unit (NICU), it was subsequently noted that very low birth weight (VLBW) infants improved clinically, despite subsequently positive blood cultures for oxacillin-resistant, coagulase-negative Staphylococcus (CONS). The results of the management of VLBW infants with CONS sepsis during one calendar year, based on clinical rather than laboratory findings, are presented. VLBW infants with LOS were identified through the neonatal database, and the charts of those with CONS were reviewed for antibiotic usage, antibiotic resistance pattern, clearance of CONS from the blood and NICU discharge status. Oxacillin sensitivity was determined by the presence of the mecA gene. From January 1 to December 31, 2002, 27 VLBW infants, treated in the authors' NICU for LOS due to CONS, were identified. The mean age of LOS infants with CONS was 15 days (median 12 days; range three to 54 days), the mean birth weight (+/- SD) was 904+/-247 g, and the mean gestational age at birth (+/- SD) was 27+/-2 weeks. All infants were started on cloxacillin and gentamicin, and improved clinically over the first 48 h. Six isolates were sensitive to cloxacillin. Twenty-three infants grew oxacillin-resistant CONS, eight of whom had persistence of CONS on repeat culture secondary to central lines. Two infants grew two strains of CONS. Eighteen of 22 infants (82%) with in vitro oxacillin-resistant CONS had clearance of bacteremia with cloxacillin and gentamicin. Ten infants (37%) received vancomycin, based on the authors' guidelines. There were no cases of prolonged bacteremia requiring rifampicin. Three infants died, but none of the deaths could be attributed to CONS. The authors describe clinical improvement with clearance of CONS using cloxacillin and gentamicin, despite laboratory results suggesting oxacillin resistance. The authors' unit
Cristina García de la Mària
Full Text Available This study describes coagulase-negative staphylococcal (CoNS infective endocarditis (IE epidemiology at our institution, the antibiotic susceptibility profile, and the influence of vancomycin minimum inhibitory concentration (MIC on patient outcomes. One hundred and three adults with definite IE admitted to an 850-bed tertiary care hospital in Barcelona from 1995-2008 were prospectively included in the cohort. We observed that CoNS IE was an important cause of community-acquired and healthcare-associated IE; one-third of patients involved native valves. Staphylococcus epidermidis was the most frequent species, methicillin-resistant in 52% of patients. CoNS frozen isolates were available in 88 patients. Vancomycin MICs of 2.0 μg/mL were common; almost all cases were found among S. epidermidis isolates and did not increase over time. Eighty-five patients were treated either with cloxacillin or vancomycin: 38 patients (Group 1 were treated with cloxacillin, and 47 received vancomycin; of these 47, 27 had CoNS isolates with a vancomycin MIC <2.0 μg/mL (Group 2, 20 had isolates with a vancomycin MIC ≥ 2.0 μg/mL (Group 3. One-year mortality was 21%, 48%, and 65% in Groups 1, 2, and 3, respectively (P = 0.003. After adjusting for confounders and taking Group 2 as a reference, methicillin-susceptibility was associated with lower 1-year mortality (OR 0.12, 95% CI 0.02-0.55, and vancomycin MIC ≥ 2.0 μg/mL showed a trend to higher 1-year mortality (OR 3.7, 95% CI 0.9-15.2; P=0.069. Other independent variables associated with 1-year mortality were heart failure (OR 6.2, 95% CI 1.5-25.2 and pacemaker lead IE (OR 0.1, 95%CI 0.02-0.51. In conclusion, methicillin-resistant S.epidermidis was the leading cause of CoNS IE, and patients receiving vancomycin had higher mortality rates than those receiving cloxacillin; mortality was higher among patients having isolates with vancomycin MICs ≥ 2.0 μg/mL.
Christina W Obiero
Full Text Available Bloodstream infection is a common cause of morbidity in children aged <5 years in developing countries. In studies reporting bacteremia in Africa, coagulase-negative Staphylococci (CoNS are commonly isolated. However, it is currently unclear whether children who are highly susceptible to infection because of severe acute malnutrition (SAM or HIV should be treated with antimicrobials specifically to cover CoNS. We aimed to determine the clinical significance of CoNS amongst children admitted to a rural hospital in Kenya in relation to nutritional and HIV status.Systematically collected clinical and microbiological surveillance data from children aged 6-59 months admitted to Kilifi County Hospital (2007-2013 were analysed. Multivariable regression was used to test associations between CoNS isolation from blood cultures and SAM (MUAC <11.5cm or nutritional oedema (kwashiorkor, and HIV serostatus; and among children with SAM or HIV, associations between CoNS isolation and mortality, duration of hospitalization and clinical features.CoNS were isolated from blood culture in 906/13,315 (6.8% children, of whom 135/906 (14.9% had SAM and 54/906 (6.0% were HIV antibody positive. CoNS isolation was not associated with SAM (MUAC<11.5cm (aOR 1.11, 95% CI 0.88-1.40 or kwashiorkor (aOR 0.84, 95% CI 0.48-1.49, or a positive HIV antibody test (aOR 1.25, 95% CI 0.92-1.71. Among children with SAM or a positive HIV antibody test, CoNS isolation was not associated with mortality or prolonged hospitalization.In a large, systematic study, there was no evidence that antimicrobial therapy should specifically target CoNS amongst children with SAM or HIV-infection or exposure.
Martínez-Meléndez, Adrián; Morfín-Otero, Rayo; Villarreal-Treviño, Licet; Camacho-Ortíz, Adrián; González-González, Gloria; Llaca-Díaz, Jorge; Rodríguez-Noriega, Eduardo; Garza-González, Elvira
The mechanisms contributing to persistence of coagulase-negative staphylococci are diverse; to better understanding of their dynamics, the characterization of nosocomial isolates is needed. Our aim was to characterize phenotypic and molecular characteristics of Staphylococcus epidermidis and Staphylococcus haemolyticus human blood isolates from two tertiary care hospitals in Mexico, the Hospital Universitario in Monterrey and the Hospital Civil in Guadalajara. Antimicrobial susceptibility was determined. Biofilm formation was assessed by crystal violet staining. Detection of the ica operon and Staphylococcal Cassette Chromosome mec typing were performed by PCR. Clonal relatedness was determined by Pulsed-fiel gel electrophoresis and Multi locus sequence typing. Methicillin-resistance was 85.5% and 93.2% for S. epidermidis and S. haemolyticus, respectively. Both species showed resistance >70% to norfloxacin, clindamycin, levofloxacin, trimethoprim/sulfamethoxazole, and erythromycin. Three S. epidermidis and two S. haemolyticus isolates were linezolid-resistant (one isolate of each species was cfr+). Most isolates of both species were strong biofilm producers (92.8% of S. epidermidis and 72.9% of S. haemolyticus). The ica operon was amplified in 36 (43.4%) S. epidermidis isolates. SCCmec type IV was found in 47.2% of the S. epidermidis isolates and SCCmec type V in 14.5% of S. haemolyticus isolates. No clonal relatedness was found in either species. Resistance to clindamycin, levofloxacin, erythromycin, oxacillin, and cefoxitin was associated with biofilm production for both species (pepidermidis isolates belonged to ST23; isolate with SCCmec type IV belonged to ST7, and isolate with SCCmec type III belonged to ST2. This is the first report of ST7 in Mexico. There was a high genetic diversity in both species, though both species shared characteristics that may contibute to virulence. Copyright © 2016 Elsevier Editora Ltda. All rights reserved.
Blanco, Claudia; Núñez, María Ximena
To determine species of staphylococci in chronic conjunctivitis, their antibiotic susceptibility pattern, patient treatments, clinical course, and clinical conditions. In this prospective study, 243 conjunctival cultures were taken from 191 patients with chronic conjunctivitis, we obtained staphylococci susceptibility patterns with E-test, and they were analyzed in coagulase-positive and negative. The minimum inhibitory concentration for 90% of isolates (MIC90) was determined for Staphylococcus aureus and Staphylococcus epidermidis. Additionally, clinical follow-up and associated factors of all patients were analyzed depending on methicillin resistance (MR) or susceptibility (MS) bacterial state. One hundred and eight (44%) cultures were positive; 81 positive cultures were Gram-positive of which, 77 were staphylococci, 29 coagulase-positive with S. aureus as the most prevalent, 89% MS, and 11% MR. And 48 were coagulase-negative with S. epidermidis as the most isolated with 36% of MS and 64% of MR. Poor susceptibility was found in the staphylococcus coagulase-negative/MR group. Moxifloxacin and vancomycin show the best in vitro activity for all isolates. The MIC90 of moxifloxacin and vancomycin were 0.064/1.5, 0.64/3.0, and 1/3.0 for S. aureus-MS, S. epidermidis-MS, and S. epidermidis-MR, respectively. The most frequently associated factors found in patients with positive culture for staphylococcus were exposure to the health care system 23 (29.87%) of 77 patients and dry eye 23 (29.87%) of 77 patients. Both with a proportion of 3 in 10. Coagulase-negative staphylococci were the most frequently isolated from the conjunctiva with 58.33% of MR; even though multiresistance was detected, their susceptibility to a fourth-generation fluoroquinolone, commonly used, such as moxifloxacin, was preserved.
Bansal, B K; Gupta, D K; Shafi, T A; Sharma, S
...) 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...
Jarløv, J O; Rosdahl, V T; Mortensen, I
-level of the isoxazolyl penicillins showed a high degree of uniformity. However more strains were resistant to cloxacillin and oxacillin than to dicloxacillin and flucloxacillin. Only a weak correlation was found between beta-lactamase production, and resistance to the six antibiotics. Methicillin was the most stable...
Renneberg, J; Rieneck, K; Gutschik, E
The purpose of the investigation was to evaluate two commercially available identification systems: a new modification of the Staph-Zym system (Rosco, Tåstrup, Denmark) and the Staph ID 32 API system (API System, BioMérieux, Paris, France). A local standard method to be used in routine laboratories was also evaluated. A total of 200 staphylococcal isolates, including strains from both the American Type Culture Collection and the Czechoslovak Collection of Microorganisms as well as 89 clinical...
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.
Romanelli, Roberta Maia de Castro; Anchieta, Lêni Márcia; Bueno E Silva, Ana Carolina; de Jesus, Lenize Adriana; Rosado, Viviane; Clemente, Wanessa Trindade
The aim of this study was to compare two different empiric treatments for late-onset neonatal sepsis, vancomycin and oxacillin, in a neonatal intensive care unit with a high prevalence of coagulase-negative Staphylococcus. A cross-sectional study was conducted in an neonatal intensive care unit from 2011 to 2014. Data from the medical records of at-risk newborns were collected daily. Infections were defined according to the National Health Surveillance Agency criteria. Data analysis was performed using an internal program. There was a significant reduction in the number of Staphylococcus aureus infections (p=0.008), without endocarditis, meningitis, or lower respiratory tract infection, as well as a reduction in the frequency of deaths related to S. aureus infection. There were no significant changes in the incidence of Gram-negative bacterial or fungal infections. An increase in coagulase-negative Staphylococcus infections was observed (p=0.022). However, there was no measured increase in related morbidity and mortality. There was a reduction in the median number of days of treatment with oxacillin from 11.5 to 6 days (pneonatal late-onset sepsis with use of oxacillin showed a significant reduction in S. aureus infections, as well as a reduction in the frequency of infections with major organ system involvement and mortality due to infection with this microorganism. As a result, oxacillin can be considered as an effective treatment for late-onset sepsis, making it possible to avoid broad-spectrum antibiotics. Copyright © 2016. Published by Elsevier Editora Ltda.
Roberta Maia de Castro Romanelli
Full Text Available Abstract Objective: The aim of this study was to compare two different empiric treatments for late-onset neonatal sepsis, vancomycin and oxacillin, in a neonatal intensive care unit with a high prevalence of coagulase-negative Staphylococcus. Methods: A cross-sectional study was conducted in an neonatal intensive care unit from 2011 to 2014. Data from the medical records of at-risk newborns were collected daily. Infections were defined according to the National Health Surveillance Agency criteria. Data analysis was performed using an internal program. Results: There was a significant reduction in the number of Staphylococcus aureus infections (p = 0.008, without endocarditis, meningitis, or lower respiratory tract infection, as well as a reduction in the frequency of deaths related to S. aureus infection. There were no significant changes in the incidence of Gram-negative bacterial or fungal infections. An increase in coagulase-negative Staphylococcus infections was observed (p = 0.022. However, there was no measured increase in related morbidity and mortality. There was a reduction in the median number of days of treatment with oxacillin from 11.5 to 6 days (p < 0.001 and an increase of one day in the median number of days of treatment with vancomycin (p = 0.046. Conclusions: Modification of the empiric treatment regimen for neonatal late-onset sepsis with use of oxacillin showed a significant reduction in S. aureus infections, as well as a reduction in the frequency of infections with major organ system involvement and mortality due to infection with this microorganism. As a result, oxacillin can be considered as an effective treatment for late-onset sepsis, making it possible to avoid broad-spectrum antibiotics.
Caracterização fenotípica da resistência a antimicrobianos e detecção do gene mecA em Staphylococcus spp. coagulase-negativos isolados de amostras animais e humanas Phenotypic characterization of antimicrobial resistance and detection of the mecA gene in coagulase-negative Staphylococcus spp. isolates from animal and human samples
Lidiane de Castro Soares
Full Text Available Os estafilococos coagulase-negativos (ECN fazem parte da microbiota normal da pele e, apesar de terem sido considerados saprófitas por muito tempo, o seu significado clínico como agente etiológico tem aumentado com o passar dos anos. Neste estudo, foram obtidos 72 isolados de ECN a partir de amostras do conduto auditivo de cães, de mastite bovina e de infecções humanas. Staphylococcus xylosus foi o microrganismo mais isolado, nas amostras animais, e S. cohnii subsp. cohnii em humanos. Os isolados foram avaliados de modo a traçar o perfil fenotípico de sua resistência aos antimicrobianos mais indicados no tratamento de infecções estafilocócicas. Foi detectado um elevado nível de resistência à penicilina e ampicilina. A gentamicina, a vancomicina e a associação ampicilina+sulbactam foram eficientes frente aos isolados testados. A resistência à oxacilina foi avaliada por meio dos testes de difusão em disco modificada, ágar screen, microdiluição em caldo e diluição em ágar para constatar, se à semelhança do que ocorre com os estafilococos coagulase-positivo, esta pode ser mediada pelo gene mecA e apresentada de forma heterogênea. A presença do gene mecA foi determinada pelo método da Reação em Cadeia de Polimerase (PCR, sendo 5,6% dos isolados mecA positivos.Coagulase-negative staphylococci (SCN make part of the normal microbiota skin and although they have been considered saprophytics for years, nowadays their clinical significance as an etiologic agent has increased. In this study, 72 SCN isolates obtained from external ear canals of dogs, bovine mastitis and human nosocomial infections were evaluated. Staphylococcus xylosus was the most prevalent microorganism in animal samples and S. cohnii subsp. cohnii in human samples. SCN isolates were evaluated in order to establish a phenotypical resistance pattern towards the most indicated antibiotics for staphyloccocal infections. A high level of resistance to penicillin
Chu, V. H.; Miro, J. M.; Hoen, B.; Cabell, C. H.; Pappas, P. A.; Jones, P.; Stryjewski, M. E.; Anguera, I.; Braun, S.; Muñoz, P.; Commerford, P.; Tornos, P.; Francis, J.; Oyonarte, M.; Selton-Suty, C.; Morris, A. J.; Habib, G.; Almirante, B.; Sexton, D. J.; Corey, G. R.; Fowler, V. G.; Gordon, David; Devi, Uma; Spelman, Denis; van der Meer, Jan T. M.; Kauffman, Carol; Bradley, Suzanne; Armstrong, William; Giannitsioti, Efthymia; Giamarellou, Helen; Lerakis, Stamatios; del Rio, Ana; Moreno, Asuncion; Mestres, Carlos A.; Pare, Carlos; Garcia de la Maria, Cristina; Claramonte, Xavier; Armero, Yolanda; de Lazzario, Elisa; Marco, Francesc; Gatell, Jose M.; Miro, José M.; Almela, Manel; Azqueta, Manuel; Jiménez-Expósito, Maria Jesús; de Benito, Natividad; Ramirez, Jose; Perez, Noel; Almirante, Benito; Fernandez-Hidalgo, Nuria; Rodriguez de Vera, Pablo; Tornos, Pilar; Falcó, Vicente; Sidani, Nisreen; Kanj-Sharara, Souha; Kanafani, Zeina; Raglio, Annibale; Goglio, Antonio; Gnecchi, Fabrizio; Suter, Fredy; Valsecchi, Grazia; Rizzi, Marco; Ravasio, Veronica; Hoen, Bruno; Chirouze, Catherine; Leroy, Joel; Plesiat, Patrick; Bernard, Yvette; Casey, Anna; Lambert, Peter; Watkin, Richard; Elliott, Tom; Patel, Mukesh; Dismukes, William; Pan, Angelo; Caros, Giampiero; Tribouilloy, Amel Brahim Mathiron Christophe; Goissen, Thomas; Delahaye, Armelle; Delahaye, Francois; Vandenesch, Francois; Vizzotti, Carla; Nacinovich, Francisco M.; Marin, Marcelo; Trivi, Marcelo; Lombardero, Martin; Cortes, Claudia; Casabe, José Horacio; Altclas, Javier; Kogan, Silvia; Clara, Liliana; Sanchez, Marisa; Commerford, Anita; Hansa, Cass; Deetlefs, Eduan; Ntsekhe, Mpiko; Commerford, Patrick; Wray, Dannah; Steed, Lisa L.; Church, Preston; Cantey, Robert; Morris, Arthur; Holland, David; Murdoch, David; Read, Kerry; Raymond, Nigel; Lang, Selwyn; Chambers, Stephen; Kotsanas, Despina; Korman, Tony M.; Peterson, Gail; Purcell, Jon; Southern, Paul M.; Shah, Manisha; Bedimo, Roger; Reddy, Arjun; Levine, Donald; Dhar, Gaurav; Hanlon-Feeney, Alanna; Hannan, Margaret; Kelly, Sinead; Wang, Andrew; Cabell, Christopher H.; Woods, Christopher W.; Sexton, Daniel J.; Benjamin, Danny; Corey, G. Ralph; McDonald, Jay R.; Federspiel, Jeff; Engemann, John J.; Reller, L. Barth; Drew, Laura; Caram, L. B.; Stryjewski, Martin; Morpeth, Susan; Lalani, Tahaniyat; Fowler, Vance; Chu, Vivian; Mazaheri, Bahram; Neuerburg, Carl; Naber, Christoph; Athan, Eugene; Henry, Margaret; Harris, Owen; Alestig, Eric; Olaison, Lars; Wikstrom, Lotta; Snygg-Martin, Ulrika; Francis, Johnson; Venugopal, K.; Nair, Lathi; Thomas, Vinod; Chaiworramukkun, Jaruwan; Pachirat, Orathai; Chetchotisakd, Ploenchan; Suwanich, Tewan; Kamarulzaman, Adeeba; Tamin, Syahidah Syed; Premru, Manica Mueller; Logar, Mateja; Lejko-Zupanc, Tatjana; Orezzi, Christina; Klein, John; Bouza, Emilio; Moreno, Mar; Rodríguez-Créixems, Marta; Marín, Mercedes; Fernández, Miguel; Muñoz, Patricia; Fernández, Rocío; Ramallo, Victor; Raoult, Didier; Thuny, Franck; Habib, Gilbert; Casalta, Jean-Paul; Fournier, Pierre-Edouard; Chipigina, Natalia; Kirill, Ozerecky; Vinogradova, Tatiana; Kulichenko, Vadim P.; Butkevich, O. M.; Lion, Christine; Selton-Suty, Christine; Alla, Francois; Coyard, Hélène; Doco-Lecompte, Thanh; Iarussi, Diana; Durante-Mangoni, Emanuele; Ragone, Enrico; Dialetto, Giovanni; Tripodi, Marie Françoise; Utili, Riccardo; Casillo, Roberta; Kumar, A. Sampath; Sharma, Gautam; Dickerman, Stuart A.; Street, Alan; Eisen, Damon Peter; Grigg, Leeanne; Abrutyn, Elias; Michelet, Christian; Tattevin, Pierre; Donnio, Pierre Yves; Fortes, Claudio Querido; Edathodu, Jameela; Al-Hegelan, Mashael; Font, Bernat; Anguera, Ignasi; Guma, Joan Raimon; Cereceda, M.; Oyonarte, Miguel J.; Mella, Rodrigo Montagna; Garcia, Patricia; Jones, Sandra Braun; Ramos, Auristela Isabel de Oliveira; Paiva, Marcelo Goulart; Tranchesi, Regina Aparecida de Medeiros; Woon, Lok Ley; Lum, Luh-Nah; Tan, Ru-San; Rees, David; Kornecny, Pam; Lawrence, Richard; Dever, Robyn; Post, Jeffrey; Jones, Phillip; Ryan, Suzanne; Harkness, John; Feneley, Michael; Rubinstein, Ethan; Strahilewitz, Jacob; Ionac, Adina; Mornos, Cristian; Dragulescu, Stefan; Forno, Davide; Cecchi, Enrico; de Rosa, Francesco; Imazio, Massimo; Trinchero, Rita; Wiesbauer, Franz; Gattringer, Rainer; Deans, Greg; Andrasevic, Arjana Tambic; Barsic, Bruno; Klinar, Igor; Vincelj, Josip; Bukovski, Suzana; Krajinovic, Vladimir
OBJECTIVE: To describe the contemporary features of coagulase-negative staphylococcal (CoNS) prosthetic valve endocarditis (PVE). DESIGN: Observational study of prospectively collected data from a multinational cohort of patients with infective endocarditis. Patients with CoNS PVE were compared to
Mossel, D. A. A.
Mossel, D. A. A. (Central Institute for Nutrition and Food Research T.N.O., Utrecht, The Nethrlands). Attempt in classification of catalase-positive staphylococci and micrococci. J. Bacteriol. 84:1140–1147. 1962.—About 390 strains of Staphylococcus aureus, isolated from clinical material, and about 190 strains of coagulase-negative staphylococci and micrococci from strictly nonclinical habitats were studied by the following recently recommended biochemical tests: anaerobic dissimilation (“fermentation”) of mannitol, gelatin liquefaction, type of growth on tellurite-glycine agar, hydrolysis of urea, and KCN tolerance. The latter three tests appeared either not specific for, or not positive for, most S. aureus strains. Virtually all strains of S. aureus were gelatin-positive, but 71% of the other types of cocci also liquefied gelatin. Rapid anaerobic breakdown of mannitol, however, was shown by ca. 95% of the strains of S. aureus, and late fermentation by an additional 3%. Of 105 obligately aerobic coagulase tive cocci (micrococci), none fermented mannitol; of 40 facultatively anaerobic, coagulase-negative cocci (staphylococci), only 7 (18%) fermented mannitol. Oxidative metabolism of mannitol occurred in only three (mannitol salt agar while 13% of the S. aureus strains tested were white. A key to the classification of catalase-positive cocci consistent with that currently used for Enterobacteriaceae has been based on these figures. PMID:13936221
López, Yolanda; Samudio, Margarita; Fariña, Norma; Castillo, Verónica; Abente, Sonia; Nentwich, Martin M; González-Britez, Nilsa; Laspina, Florentina; Carron, Agustín; Cibils, Diógenes; de Kaspar, Herminia Miño
In this prospective study, multiplex polymerase chain reaction (PCR) was used to identify genes encoding virulence factors (ica, atlE and mecA) in Coagulase-negative Staphylococcus (CNS) isolates from the ocular microbiota of patients undergoing cataract surgery and to investigate possible changes in the CNS profile due to antibiotic prophylaxis. Between 09/2011 and 08/2013, patients undergoing cataract surgery were recruited at the Department of Ophthalmology, National University of Asuncion, Paraguay. In the eye to be operated on, patients received moxifloxacin 0.5 % eye drops four times at the day before surgery and a last drop 1 hour before surgery (T1). The other eye remained as control (T0). Conjunctival swabs were taken from both eyes 1 hour after the last drop. The presence of genes encoding biofilm formation (ica and atlE) and methicillin resistance (mecA) was detected by a multiplex PCR. Of the 162 patients (162 study eyes, 162 fellow eye as control group), 87 (53.7 %) eyes were positive for CNS at T0 yielding 96 CNS isolates; 70 eyes (43.2 %) were positive at T1 yielding 77 CNS isolates. For this study, 43 CNS isolates (44.8 %) from T0 and 45 (64.3 %) from T1 were used. Of the total isolates, 81.8 % (72/88) had at least one virulence factor gene (37/43 from T0 and 35/45 from T1) (p = 0.314). Simultaneous detection of ica and atlE genes was higher in T0 (58.0 %) than T1 (46.7 %), but the difference was not significant (p = 0.28). A high frequency of genes encoding virulence factors was observed in the coagulase-negative Staphylococcus isolates. The use of moxifloxacin did not significantly modify the CNS virulence factor profiles.
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.
Yazgi, H; Uyanik, M H; Ayyildiz, A
This study investigated the colonization of slime-producing coagulase-negative Staphylococcus (CoNS) in 80 patient wards in Turkey (40 vinyl and 40 ceramic tile floors). A total of 480 samples that included 557 CoNS isolates were obtained. Slime production was investigated with the Christensen method and methicillin-susceptibility was tested by the disk-diffusion method. There was a significant difference in the percentage of slime-producing CoNS isolates on vinyl (12.4%) versus ceramic tile flooring (4.4%). From vinyl flooring, the percentage of slime producing methicillin-resistant CoNS (MRCoNS) (8.9%) was significantly higher than for methicillin-sensitive CoNS (MSCoNS) (3.6%), whereas there was no difference from ceramic tile flooring (2.5% MRCoNS versus 1.8% MSCoNS). The most commonly isolated slime-producing CoNS species was S. epidermidis on both types of flooring. It is concluded that vinyl flooring seems to be a more suitable colonization surface for slime-producing CoNS than ceramic tile floors. Further studies are needed to investigate bacterial strains colonized on flooring materials, which are potential pathogens for nosocomial infections.
Laurence Van Melderen
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.
Short communication: Identification of coagulase-negative staphylococcus species from goat milk with the API Staph identification test and with transfer RNA-intergenic spacer PCR combined with capillary electrophoresis.
Koop, G; De Visscher, A; Collar, C A; Bacon, D A C; Maga, E A; Murray, J D; Supré, K; De Vliegher, S; Haesebrouck, F; Rowe, J D; Nielen, M; van Werven, T
Coagulase-negative staphylococci (CNS) are the most commonly isolated bacteria from goat milk, but they have often been identified with phenotypic methods, which may have resulted in misclassification. The aims of this paper were to assess the amount of misclassification of a phenotypic test for identifying CNS species from goat milk compared with transfer RNA intergenic spacer PCR (tDNA-PCR) followed by capillary electrophoresis, and to apply the tDNA-PCR technique on different capillary electrophoresis equipment. Milk samples were collected from 416 does in 5 Californian dairy goat herds on 3 occasions during lactation. In total, 219 CNS isolates were identified at the species level with tDNA-PCR and subjected to the API 20 Staph identification test kit (API Staph; bioMérieux, Durham, NC). If the same species was isolated multiple times from the same udder gland, only the first isolate was used for further analyses, resulting in 115 unique CNS isolates. According to the tDNA-PCR test, the most prevalent CNS species were Staphylococcus epidermidis, Staphylococcus caprae, and Staphylococcus simulans. Typeability with API staph was low (72%). Although the API Staph test was capable of identifying the majority of Staph. epidermidis and Staph. caprae isolates, sensitivity for identification of Staph. simulans was low. The true positive fraction was high for the 3 most prevalent species. It was concluded that the overall performance of API Staph in differentiating CNS species from goat milk was moderate to low, mainly because of the low typeability, and that genotypic methods such as tDNA-PCR are preferred. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
Full Text Available Introduction: Staphylococci release a large number of enzymes. Some of these, such as coagulase, beta lactamase, hemolysins and biofilms are considered indices of pathogenicity. The aim of the current study was based on the isolation and identification of Staphylococcus aureus and coagulase negative Staphylococci (CNS strains from sheep sub clinical mastitis and examining their biofilm, beta lactamase, hemolysins production and antibiotic resistance pattern. Materials and methods: 55 Staphylococci strains were isolated from seventy cases of sheep subclinical mastitis. Thirty three were determined as Staphylococcus aureus (60% and 22 (40% as CNS. The hemolytic activity was evaluated by plating Staphylococci strains on 5% bovine blood agar. The biofilm assay was performed by using micro titer plates. Beta Lactamase production was detected by test tube iodometric technique and susceptibility to antimicrobial agents was determined for isolated strains by the disk diffusion method. Results: Twenty six (78.8% S. aureus strains were biofilm producers. For CNS (59.9% strains were positive in biofilm production. Two isolates (6.06%, of S. aureus were α, the same number β and 6 (18.2% isolates were ∂ hemolysin producers. Six isolates of CNS (27.27% were α and ten (45.45% ∂ hemolysin producers. Sixteen S. aureus (48.5% and five CNS (22.72% isolates were positive in beta lactamase production. The isolated Staphylococci show a low sensitivity pattern to methicillin and streptomycin. Discussion and conclusion: A high percentage of strains make α toxin that play a role in S. aureus biofilm formation. Twenty one out of 33 (63.63% isolated Staphylococci were biofilm producers that can have deleterious effects because biofilm formation is thought to play an important role in the survival of virulent strains of Staphylococci. Sixteen out of 33 (48.5% isolated S. aureus were positive in beta lactamase test, Excluding resistant to methicillin, all of these
Full Text Available A total of 187 isolates from several clinical specimens were identified to species level as 129 Staphylococcus aureus strains and 58 coagulase-negative staphylococci (CNS strains by the API Staph System (Biomerieux. Slime production was detected both by the conventional Christensen's method as well as by the Congo red agar method. Seventy-two strains of staphylococci isolates (38.5% were found to be slime producers by Christensen's test tube method whereas 58 strains (31% were slime positive with Congo red agar method. There was no statistically significant difference between the two methods for the detection of slime production (P > 0.05. Susceptibility of isolates against antimicrobial agents was tested by the disk diffusion method. Staphylococcal species had resistance to one or more antibiotics. Among the various antimicrobial agents, oxacillin (71.1% and erythromycin (47.1% showed higher resistance than most of the agents used against all isolates. Oxacillin resistant S. aureus (ORSA and oxacillin resistant coagulase-negative staphylococci (ORCNS, 97 (75.2% and 36 (62.1% respectively were frequently observed in strains isolated from clinical materials. Among the ORSA strains, two strains were resistant to vancomycin. Moreover, 96 (74.4% of 129 S. aureus strains were positive for blactamase enzyme. However, 78 (81.25% of 96 b-lactamase positive S. aureus strains were b-lactamase positive ORSA isolates, but none of them had vancomycin resistance.
Okolie, Charles E
The genus Staphylococcus includes pathogenic and non-pathogenic facultative anaerobes. Due to the plethora of virulence factors encoded in its genome, the species Staphylococcus aureus is known to be the most pathogenic. S. aureus strains harboring genes encoding virulence and antibiotic resistance are of public health importance. In clinical samples, however, pathogenic S. aureus is often mixed with putatively less pathogenic coagulase-negative staphylococci (CoNS), both of which can harbor mecA, the genetic driver for staphylococcal methicillin-resistance. In this chapter, the detailed practical procedure for operating a real-time pentaplex PCR assay in blood cultures is described. The pentaplex real-time PCR assay simultaneously detects markers for the presence of bacteria (16S rRNA), coagulase-negative staphylococcus (cns), S. aureus (spa), Panton-Valentine leukocidin (pvl), and methicillin resistance (mecA).
Stepanović, S; Cirković, I; Djukić, S; Vuković, D; Svabić-Vlahović, M
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.
Planchon, Stella; Desvaux, Mickaël; Chafsey, Ingrid; Chambon, Christophe; Leroy, Sabine; Hébraud, Michel; Talon, Régine
Staphylococcus xylosus is a Gram-positive bacterium found on the skin of mammals and frequently isolated from food plants and fermented cheese or meat. To gain further insight in protein determinants involved in biofilm formation by this coagulase-negative Staphylococcus, a comparative proteomic analysis between planktonic and sessile cells was performed. With the use of a protocol previously developed, protein patterns of the cytoplasmic and cell envelope fractions were compared by 2-DE. Following protein identification by MALDI-TOF mass spectrometry and bioinformatic analyses, this study revealed differences in expression levels of 89 distinct proteins with 55 up-expressed and 34 down-expressed proteins in biofilm compared to planktonic cells. Most proteins differentially expressed were related to nitrogen and carbon metabolisms. Besides amino acid biosynthesis and protein translation, protein determinants related to protein secretion were up-expressed in biofilm, suggesting a more active protein trafficking in sessile cells. While up-expression of several enzymes involved in pentose phosphate and glycolytic pathways was observed in biofilm, connections with unexpected metabolic routes were further unravelled. Indeed, this proteomic analysis allowed identifying novel proteins that could be involved in a previously uncovered exopolysaccharide biosynthetic pathway in S. xylosus as well as several enzymes related to polyketide biosynthesis. This findings are particularly relevant considering exopolysaccharide production in S. xylosus is ica-independent contrary to coagulase-negative model strain Staphylococcus epidermidis RP62A.
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
Outbreaks associated to bloodstream infections with Staphylococcus aureus and coagulase-negative Staphylococcus spp in premature neonates in a university hospital from Brazil Surtos associados a infecções sanguíneas por Staphylococcus aureus e Staphylococcus spp coagulase-negativa em neonatos prematuros em um hospital universitário do Brasil
Denise Von Dolinger de Brito
Full Text Available Staphylococcus aureus and coagulase-negative staphylococci (CoNS are among the most important nosocomial pathogens in patients from neonatal intensive care units, mainly in bloodstream infections. The main objective of this study is to determine the occurrence of nosocomial infections by these microorganisms using two surveillance systems (Laboratorial Surveillance and National Nosocomial Infection Surveillance System and to determine the most important risk factors during a two-year period (2001-2002. Two outbreaks by both methicillin susceptible S. aureus (MSSA (1.5% and methicillin resistant CoNS (MRCoNS (1.0% were observed, from January to February/02 and August to September/02. Endemic incidence rates of 3.77% and 5.16% of S. aureus and CoNS, respectively were detected. Risk factors included age or = 7 days and utilization of polietilene central vascular catheter (CVC through vein dissection (phlebotomy, but none of these independent factors were confirmed by the multivariate analysis. However, oxacillin resistant CoNS prevailed (66.0% in the epidemic episodes. Molecular analysis by pulsed field gel electrophoresis showed the polyclonal nature of S. aureus isolates. In conclusion, two outbreaks were identified of mixed etiology by MSSA and MRCoNS associated to the lack of an adequate material (central venous catheter for neonates, related invasive procedure. The outbreaks were controlled with the substitution of polietilene CVC for peripherally inserted central catheter.Staphylococcus aureus e Estafilococos coagulase-negativa (ECN estão entre os patógenos hospitalares mais importantes em pacientes de unidades de terapia intensiva neonatal, principalmente em infecções da corrente sanguínea. O principal objetivo deste estudo foi determinar a ocorrência de infecções hospitalares por estes microrganismos usando dois sistemas de vigilância (laboratorial e "National Nosocomial Infection Surveillance" - NNIS e determinar os fatores de
Bagcigil, Funda A.; Moodley, Arshnee; Baptiste, Keith E.
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...
Taponen, S.; Simojoki, H.; Haveri, M.
-typing of CNS was compared with the phenotypic identification. A total of 133 clinical or subclinical quarter cases of intramammary infection caused by CNS from the practice area of the Ambulatory Clinic of the University of Helsinki were studied. Bacteriological diagnosis was based on biochemical (API) testing....... Staphylococcus simulans (43.6%) followed by S. chromogenes (23.3%) were the most common CNS species isolated from the milk samples. Ninety-nine isolates were genotyped using AFLP-analysis. Only 75.0% of S. chromogenes and S. simulans isolates identified with API test were clustered with the type strains...
Full Text Available We analyzed the occurrence and mechanisms of fusidic acid resistance present in staphylococci isolated from 59 healthy volunteers. The fingers of the volunteers were screened for the presence of staphylococci, and the collected isolates were tested for resistance to fusidic acid. A total of 34 fusidic acid resistant staphylococcal strains (all were coagulase-negative were isolated from 22 individuals (22/59, 37.3%. Examination of the resistance genes revealed that acquired fusB or fusC was present in Staphylococcus epidermidis, Staphylococcus capitis subsp. urealyticus, Staphylococcus hominis subsp. hominis, Staphylococcus warneri and Staphylococcus haemolyticus. Resistance islands (RIs carrying fusB were found in S. epidermidis and S. capitis subsp. urealyticus, while staphylococcal chromosome cassette (SCC-related structures harboring fusC were found in S. hominis subsp. hominis. Genotypic analysis of S. epidermidis and S. hominis subsp. hominis indicated that the fus elements were disseminated in diverse genetic strain backgrounds. The fusC elements in S. hominis subsp. hominis strains were highly homologous to SCCfusC in the epidemic sequence type (ST 239/SCCmecIII methicillin-resistant S. aureus (MRSA or the pseudo SCCmec in ST779 MRSA. The presence of acquired fusidic acid resistance genes and their genetic environment in commensal staphylococci suggested that the skin commensal staphylococci may act as reservoir for fusidic acid resistance genes.
Lilenbaum, Walter; Moraes, Ismar A; Cardoso, Verônica S; Varges, Renato G; Ferreira, Ana Maria R; Pissinatti, Alcides
The purpose of this study was to provide current data on Staphylococcus species from the vaginas of clinically normal captive lion tamarins and to determine the antimicrobial susceptibility of these isolates. Samples were collected from 25 adult lion tamarins, processed to isolate Staphylococcus species, and tested for susceptibility to penicillin G, gentamicin, chloramphenicol, tetracycline, trimethoprim-sulfamethoxazole, streptomycin, ampicillin, and rifampicin. Isolates with the typical characteristics of the genus Staphylococcus were recovered from all 25 samples. Coagulase-negative species were the most common (68% of the isolates), and the most frequently isolated species (10 samples) was S. simulans. Other coagulase-negative species, including S. saprophyticus (n=5), S. epidermidis (n=1), and S. arlettae (n=1), were also recovered. Coagulase-positive Staphylococci were obtained from eight animals (six of from the S. aureus species and two from S. intermedius). Resistance to antibiotics was frequently observed, and 88% of the isolates (23 samples) showed resistance to at least one drug. Resistance to penicillin G was a common finding, and the most active antimicrobial agents were chloramphenicol and gentamicin. Coagulase-positive strains were more frequently resistant to antibiotics (79.7%, average=6.4 drugs) than coagulase-negative strains (38.2%, average=3.0 drugs). The high frequency of resistance observed in those isolates is surprising and very alarming. A detailed history of the use of antimicrobial drugs in these subjects did not reveal any previous exposure to any of the tested antibiotics that could justify the observed resistance rate. Copyright (c) 2006 Wiley-Liss, Inc.
Aliye Gulmez Saglam
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.
Sağlam, Aliye Gülmez; Şahin, Mitat; Çelik, Elif; Çelebi, Özgür; Akça, Doğan; Otlu, Salih
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. 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. 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. 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.
Schmidt, Vanessa M; Williams, Nicola J; Pinchbeck, Gina; Corless, Caroline E; Shaw, Stephen; McEwan, Neil; Dawson, Susan; Nuttall, Tim
Coagulase-positive (CoPS) and coagulase-negative (CoNS) staphylococci are normal commensals of the skin and mucosa, but are also opportunist pathogens. Meticillin-resistant (MR) and multidrug-resistant (MDR) isolates are increasing in human and veterinary healthcare. Healthy humans and other animals harbour a variety of staphylococci, including MR-CoPS and MR-CoNS. The main aims of the study were to characterise the population and antimicrobial resistance profiles of staphylococci from healthy non-vet visiting and non-antimicrobial treated Labrador retrievers in the UK. Nasal and perineal samples were collected from 73 Labrador retrievers; staphylococci isolated and identified using phenotypic and biochemical methods. They were also confirmed by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS), PCR of the nuc gene and PCR and sequencing of the tuf gene. Disc diffusion and minimum inhibitory concentration (MIC) susceptibility tests were determined for a range of antimicrobials. In total, 102 CoPS (S. pseudintermedius n = 91, S. aureus n = 11) and 334 CoNS isolates were detected from 99% of dogs in this study. In 52% of dogs CoNS only were detected, with both CoNS and CoPS detected in 43% dogs and CoPS only detected in 4% of dogs. Antimicrobial resistance was not common among CoPS, but at least one MDR-CoNS isolate was detected in 34% of dogs. MR-CoNS were detected from 42% of dogs but no MR-CoPS were isolated. S. epidermidis (52% of dogs) was the most common CoNS found followed by S. warneri (30%) and S. equorum (27%), with another 15 CoNS species isolated from ≤ 15% of dogs. S. pseudintermedius and S. aureus were detected in 44% and 8% of dogs respectively. MR- and MDR-CoPS were rare. However a high prevalence of MR- and MDR-CoNS were found in these dogs, even though they had no prior antimicrobial treatment or admission to veterinary premises. These findings are of concern due to the potential for opportunistic
Mehmeti, Ibrahim; Behluli, Behlul; Mestani, Mergim; Ademi, Arsim; Nes, Ingolf F; Diep, Dzung B
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.
Silva, Lívia Viganor da; Araújo, Manuela Tedesco; Santos, Kátia Regina Netto dos; Nunes, Ana Paula Ferreira
Methicillin-resistant Staphylococcus aureus (MRSA) and coagulase-negative Staphylococcus spp (CNS) are the most common pathogens that cause serious long term infections in patients. Despite the existence of new antimicrobial agents, such as linezolid, vancomycin (VAN) remains the standard therapy for the treatment of infections caused by these multidrug-resistant strains. However, the use of VAN has been associated with a high frequency of therapeutic failures in some clinical scenarios, mainly with decreasing concentration of VAN. This work aims to evaluate the synergic potential of VAN plus sulfamethoxazole/trimethoprim (SXT), VAN plus rifampin (RIF) and VAN plus imipenem (IPM) in sub-minimum inhibitory concentrations against 22 clinical strains of MRSA and CNS. The checkerboard method showed synergism of VAN/RIF and VAN/SXT against two and three of the 22 strains, respectively. The combination of VAN with IPM showed synergistic effects against 21 out of 22 strains by the E-test method. Four strains were analyzed by the time-kill curve method and synergistic activity was observed with VAN/SXT, VAN/RIF and especially VAN/IPM in sub-inhibitory concentrations. It would be interesting to determine if synergy occurs in vivo. Evidence of in vivo synergy could lead to a reduction of the standard VAN dosage or treatment time.
Full Text Available Abstract Background Accurate identification of isolates belonging to genus Enterococcus, Streptococcus, coagulase-negative Staphylococcus, and Lactococcus at the species level is necessary to provide a better understanding of their pathogenic potential, to aid in making clinical decisions, and to conduct epidemiologic investigations,especially when large blind samples must be analyzed. It is useful to simultaneously identify species in different genera using a single primer pair. Methods We developed a primer pair based on the tuf gene (encoding elongation factor sequence to identify 56 Gram-positive cocci isolates. Results The target sequences were amplified from all 56 samples. The sequencing results and the phylogenetic tree derived from the partial tuf gene sequences identified the isolates as three enterococcal species, two lactococcal species, two staphylococcal species, and six streptococcal species, as well as eight isolates that were novel species of the genus Streptococcus. Partial gene sequence analysis of the sodA, dnaK, and 16S RNA genes confirmed the results obtained by tuf gene sequencing. Conclusion Based on the uniform amplification of the tuf gene from all samples and the ability to identify all isolates at both the genus and species levels, we conclude that the primer pair developed in this research provides a powerful tool for identifying these organisms in clinical laboratories where large blind samples are used.
Li, Xuerui; Xing, Juan; Li, Baoyu; Wang, Pu; Liu, Jixing
Accurate identification of isolates belonging to genus Enterococcus, Streptococcus, coagulase-negative Staphylococcus, and Lactococcus at the species level is necessary to provide a better understanding of their pathogenic potential, to aid in making clinical decisions, and to conduct epidemiologic investigations,especially when large blind samples must be analyzed. It is useful to simultaneously identify species in different genera using a single primer pair. We developed a primer pair based on the tuf gene (encoding elongation factor) sequence to identify 56 Gram-positive cocci isolates. The target sequences were amplified from all 56 samples. The sequencing results and the phylogenetic tree derived from the partial tuf gene sequences identified the isolates as three enterococcal species, two lactococcal species, two staphylococcal species, and six streptococcal species, as well as eight isolates that were novel species of the genus Streptococcus. Partial gene sequence analysis of the sodA, dnaK, and 16S RNA genes confirmed the results obtained by tuf gene sequencing. Based on the uniform amplification of the tuf gene from all samples and the ability to identify all isolates at both the genus and species levels, we conclude that the primer pair developed in this research provides a powerful tool for identifying these organisms in clinical laboratories where large blind samples are used.
Jay, James M.
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 α-hemolysin, 70% the δ-, 22% the β-, 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. PMID:14451457
Full Text Available Introduction: Blood culture is still the gold standard for the detection of the causative agent of sepsis. Especially in intensive care patients and those with vascular catheters, the most common organisms isolated are coagulase-negative staphylococci (CoNS and Staphylococcus aureus, both characterized by multidrug resistance. Purposes of our work are the study of the incidence of markers of resistance in staphylococci and evaluation of potential changes over the years. Materials and methods: In the period January 2008-June 2011 5239 blood cultures were analyzed.They were mainly obtained from the departments of Intensive Care, Cardiology, Hematology, General Medicine, Emergency Medicine, Infectious Diseases, Oncology, Pulmonology and Pediatric Hematoncology. The vials containing the blood were incubated in the BACTEC 9120 automated tool of Becton Dickinson and susceptibility testing performed with the Phoenix instrument of the same company. Results:Within a total of 5239 blood cultures, 3967 (75.7% were negative and 1272 (24.3% positive. Fungi were isolated in 6.2% (79 of the positive ones, Gram-negative bacteria in 24.6% (313 and Gram-positive bacteria in 69.2% (880. Within the latter, 187 (21.2% were not staphylococcal isolates, 693 (78.8% were stafiloccocci mainly represented by S. epidermidis, S. aureus, S. hominis, S. haemolyticus and S. saprophyticus. Of the 693 staphylococcal isolates, 436 (62.9% were b lactamase producers, and between them 336 (77.1% were methicillin resistant, while only 3 of 436 (0.69% were S. aureus resistant to vancomycin as well.The incidence of markers of resistance was very high, especially in patients in intensive care and cardiac surgery, who are usually subjected to combined antibiotic therapy. In the three years studied there were no statistically significant differences in the resistance of staphylococci. Conclusions: The data show an alarming high number of multi-resistant staphylococci, which is often a
[The development and testing of reagents kit for detection and qualitative evaluation of DNA of methicillin sensitive and methicillin resistant Staphylococcus aureus and also methicillin resistant coagulase negative Staphylococcus spp. applying technique of polymerase chain reaction in "real time" mode].
Skachkova, T S; Shipulina, O Iu; Domonova, É A; Subbotovskaia, A I; Kozyreva, V S; Il'ina, V N; Shipulin, G A
The reagents kit is developed to identify and quantitatively detect DNA of methicillin sensitive and methicillin resistant Staphylococcus aureus, methicillin resistant coagulase negative Staphylococcus spp. in biological material using technique of polymerase chain reaction with hybridizational fluorescent detection and having higher analytical and diagnostic characteristics. The application of the given reagents kit makes it possible to optimize the epidemiologic monitoring of propagation of methicillin resistant strains of Staphylococcus spp. Significantly decreasing duration and laboriousness of study.
Coton, Emmanuel; Mulder, Niels; Coton, Monika; Pochet, Sylvie; Trip, Hein; Lolkema, Juke S.
A multiplex PCR method, aimed at the detection of genes associated with biogenic amine production, identified the odc gene encoding ornithine decarboxylase in 1 of 15 strains of Staphylococcus epidermidis. The ability of the positive strain, S. epidermidis 2015B, to produce putrescine in vitro was demonstrated by high-performance liquid chromatography (HPLC). In this strain, the odc gene was detected on plasmid DNA, suggesting that the ability to form putrescine is carried by a mobile element, which explains the fact that the trait is strain dependent within the S. epidermidis species. A 6,292-bp nucleotide sequence harboring the putative odc gene was determined. S. epidermidis ornithine decarboxylase (ODC) showed 60 to 65% sequence identity with known ODCs of Gram-positive as well as Gram-negative bacteria. Downstream of the odc gene, a gene encoding a putative amino acid transporter was found that shared 59% sequence identity with the ornithine/putrescine exchanger (PotE) of Escherichia coli. Cloning and expression of the potE gene of S. epidermis 2015B in Lactococcus lactis demonstrated that the gene product transported ornithine and putrescine into the cells and efficiently exchanged putrescine for ornithine. Analysis of the flanking regions showed high identity levels with different S. epidermidis plasmid sequences, which would confirm the plasmidic location of the odc operon. It follows that the odc and potE gene pair encodes a putrescine-producing pathway in S. epidermis 2015B that was acquired through horizontal gene transfer. PMID:20581187
DETERMINACIÓN DEL GEN aac(6´-aph(2´´ ASOCIADO CON RESISTENCIA A AMINOGLUCÓSIDOS EN CEPAS DE STAPHYLOCOCCUS COAGULASA NEGATIVA EN UNA UNIDAD NEONATAL EN BOGOTÁ Detection of gene aac(6´-aph(2´´ in associated with aminoglucosides resistance in Staphylococcus coagulase-negative strains isolates from one neonatal unit in Bogota
Liliana Muñoz M
Full Text Available Antecedentes. La causa más común de resistencia a antibióticos aminoglucósidos en bacterias Gram positivas, especialmente en S. epidermidis, es la enzima modificante AAC(6’-APH(2", capaz de acetilar y fosforilar un amplio rango de antibióticos. Objetivo. Determinar la presencia del gen aac(6’-aph(2" en cepas de Staphylococcus coagulasa negativa aisladas en infecciones neonatales, e investigar la concordancia con las pruebas de sensibilidad in-vitro. Material y métodos. Se determinó la presencia del gen aac(6’-aph(2" en 63 cepas de estafilococos coagulasa negativa provenientes de hemocultivos y puntas de catéter, de la Unidad de Neonatología del Instituto Materno Infantil, en Bogotá. Resultados. Staphylococcus epidermidis fue el germen más frecuente y el gen aac(6’-aph(2" estaba presente en 55 (87,3% cepas; de éstas, 42 (73,4% cepas provenían de hemocultivos y 13 (23,6% cepas de punta de catéter. La susceptibilidad a gentamicina se determinó mediante concentración inhibitoria mínima y para amikacina por difusión en disco. No se encontró asociación estadísticamente significativa entre la presencia del gen aac(6’-aph(2" y la resistencia a gentamicina y amikacina. Conclusión. La presencia del gen aac(6’-aph(2" de resistencia a aminoglucósidos es muy alta en cepas de Staphylococcus epidermidis. Diferencias en la expresión del gen pueden explicar parcialmente la falta de consistencia con las pruebas de susceptibilidad utilizadas en la clínica.Background. The most common cause of resistance to aminoglucosides is the acetilation and phosphorilation of the antibiotic by the enzyme AAC(6’APH(2". Objetive. To determine the presence of the gene aac(6’-aph(2" in strains of coagulase-negative Staphylococci isolated from infected neonates and to investigate the concordance with the susceptibility tests in-vitro. Materials and methods. the aac(6’-aph(2" gene was determined in 63 coagulase-negative Staphylococci strains
Aarestrup, Frank Møller; Agersø, Yvonne; Ahrens, Peter
The species distribution, susceptibility to 19 antimicrobial agents and presence of selected genes encoding resistance to macrolides, streptogramins and tetracyclines were examined among 118 staphylococcal isolates from infections of poultry in Denmark. Isolates were identified using a combination...... 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...
Cercenado, Emilia; Morosini, María Isabel
Because of the increase in bacterial resistance, there is a need for new antimicrobial agents. In particular, Staphylococcus aureus is a frequent cause of severe infections and has an extraordinary capacity to develop antibiotic multiresistance, including resistance to glycopeptides, linezolid, and daptomycin. Although the incidence of methicillin-resistant S. aureus (MRSA) seems to have stabilized in the last few years, its wide dissemination in healthcare settings and in the community is a cause of concern. Ceftaroline is a new broad-spectrum cephalosporin with bactericidal activity against Gram-positive bacteria, including MRSA and multidrug-resistant Streptococcus pneumoniae. In addition, this drug is active against staphylococci showing resistance to glycopeptides, linezolid, and daptomycin. The ceftaroline MIC90 against MRSA ranges from 0.5-2mg/L and that against methicillin-resistant coagulase-negative staphylococci is 0.5mg/L. Ceftaroline has also good activity against respiratory pathogens including Haemophilus influenzae and Moraxella catarrhalis. Although this drug is active against Enterobacteriaceae, it does not retain activity when these isolates produce extended-spectrum beta-lactamases, carbapenemases or hyperproduce AmpC. Ceftaroline is not active against nonfermentative Gram-negative bacilli. Ceftaroline is an interesting addition to the therapeutic armamentarium against MRSA and constitutes an important option for the treatment of polymicrobial infections caused by multidrug-resistant Gram-positive microorganisms. Copyright © 2014 Elsevier España, S.L. All rights reserved.
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
Maria de Fatima Borges
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.
Ifeoma Chinyere UGWU
Full Text Available This study was conducted to isolate and detect methicillin-resistant staphylococci (MRS in healthy broilers in Nsukka Southeast, Nigeria and determine the antibiogram of the isolates. Cloacal and skin swabs were collected from each of 101 randomly sampled broilers meant for slaughter. The samples were processed for isolation and identification of methicillin-resistant Staphylococcus species, following standard methods. Confirmation of methicillin-resistance by the isolates was done using penicillin binding protein 2a (PBP2a kit. Phenotypic resistance of the isolates to antimicrobial agents was determined using disc diffusion method. Out of 202 samples processed, 200 (99.01% yielded positive growth of staphylococci on oxacillin-supplemented oxacillin-resistance staphylococcal agar base (ORSAB. A total of 200 methicillin-resistant staphylococcal isolates were obtained. Of these, 91 (45.5% were identified as methicillin-resistant coagulase-positive Staphylococcus (MRCoPS, while 109 (54.5% were identified as methicillin-resistant coagulase-negative Staphylococcus species (MRCoNS. Out of the 91 MRCoPS, 53 (58.2% were identified as methicillin-resistant Staphylococcus aureus (MRSA. Resistance of the isolates was 99.5% to erythromycin and chloramphenicol, 100% to oxacillin, 76.5% to gentamicin, 96.5% to clindamycin, 92.5% to ciprofloxacin, 99% to sulphamethoxazole/trimethoprim and tetracycline, and 98.5% to streptomycin and cefoxitin. All the isolates were multidrug resistant. This study has shown that healthy broilers reared and slaughtered in Nsukka Southeast, Nigeria harbour multidrug-resistant MRS and thus serve as their reservoirs.
Full Text Available One hundred and sixty seven isolates of staphylococci isolated from the inpatients of a tertiary care referral hospital in South India were speciated and activity of oxacillin, glycopeptides, linezolid and quinupristin/dalfopristin against these isolates was tested by broth microdilution method. Of the 114 coagulase negative staphylococci (CoNS, 49.1 % were S. haemolyticus, isolated predominantly from urine (64.6%, while the rest belonged to 11 other species. More than half the isolates of S. aureus (52.8% and 68.4% of the CoNS were oxacillin resistant. All the strains were uniformly susceptible to vancomycin, linezolid and quinupristin/dalfopristin; but 25.6% isolates of S. haemolyticus showed reduced susceptibility to teicoplanin (MIC: 8-16 mg/L. Our study demonstrates the high prevalence of oxacillin resistance among hospital isolates of S. aureus and CoNS in India. Vancomycin, along with the newer agents like linezolid and quinupristin/dalfopristin remains the drug of choice for treating multi drug resistant staphylococcal infections.
However, gene ChrA1 was shown to be present only in the genome of S. aureus. These results showed that Chr genes could be distributed from Cupriavidus metallidurans (Gram negative bacteria) to Staphylococci species (Gram positive bacteria), probably by horizontal transfer. Soixante-six Staphylococci résistants au ...
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
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.
Mallardo, Karina; Nizza, Sandra; Fiorito, Filomena; Pagnini, Ugo; De Martino, Luisa
To investigate the prevalence of methicillin-resistant staphylococci (MRS) which is a potencial risk factor of transmission between animals and humans in different types of horses (harness racing-horses, breeding mares and riding-horses) and to compare the antimicrobial resistance of the isolates. A total of 191 healthy horses, housed at different locations of the Campania Region (Italy), were included in the study. Nasal swab samples were collected from each nostril of the horses. The mecA gene was detected by a nested PCR technique. Antibiotic susceptibility was tested for each isolate. MRS was isolated from nasal samples of 68/191 (35.6%; 95% CI: 28.9%-42.9%) healthy horses. All isolates were coagulase-negative with the exception of two coagulase-positive MRS strains, identified as Staphylococcus aureus and Staphylococcus pseudintermedius, 2/83 (2.4%; 95% CI: 0.4%-9.2%). Interestingly, both coagulase-positive MRS isolates were from harness racing-horses. These horses also presented a significantly higher positivity for MRS (53.3%; 95% CI: 40.1%-66.1%) than the breeding mares and riding-horses groups. Antibiotic susceptibility testing showed difference between isolates due to different origins except for an almost common high resistance to aminopenicillins, such as ampicillin and amoxicillin. It can be concluded that harness racing-horses may act as a significant reservoir of MRS as compared to breeding mares and riding-horses.
Full Text Available Abstract Background The staphylococci are implicated in a variety of human infections; however, many clinical microbiology laboratories in Nigeria do not identify staphylococci (in particular coagulase negative staphylococci - CNS to the species level. Moreover, data from multi-centre assessment on antibiotic resistance and epidemiology of the staphylococci are not available in Nigeria. This study investigated 91 non-duplicate staphylococcal isolates obtained from the microbiology laboratories of eight hospitals in Nigeria during the period January to April 2010. Methods Identification and antibiotic susceptibility testing was performed using the VITEK 2 system, detection of resistance genes by PCR, and molecular characterization was determined by SCCmec typing, spa and multilocus sequence typing (MLST. Results All the isolates were susceptible to mupirocin, tigecycline, vancomycin and linezolid, but 72.5% of CNS and 82.3% of Staphylococcus aureus were resistant to cotrimoxazole, while multiresistance was observed in 37 of the 40 CNS isolates. Untypeable SCCmec types (ccrC/Class A mec and ccr-negative/Class C2 mec gene complex in two methicillin-resistant S. aureus (MRSA were identified. Additionally, ccr-negative/Class A mec and ccr type 4/Class C2 mec gene complex was detected in one isolate each of S. sciuri and S. haemolyticus, respectively. The S. aureus isolates were classified into 21 spa types including two new types (t8987, t9008 among the methicillin-susceptible S. aureus (MSSA isolates. Two (CC8-SCCmecnon-typeable and CC88-SCCmec IV and four (CC8-SCCmec III/IV/V; CC30-SCCmec II/III; CC88-SCCmec IV; and ST152-SCCmecnon-typeable MRSA clones were identified in Maiduguri (North-East Nigeria and South-West Nigeria, respectively. The proportion of Panton-Valentine leukocidin (PVL-positive MSSA was high (44.4% and 56.3% of these strains were associated with sequence type (ST 152. Conclusions The identification of multiresistant mecA positive
Aquino, G de V; Maluta, R P; de Ávila, F A
The aim of this work was to establish the prevalence of methicillin-resistant Staphylococci (MRS) in the animals and staff of a teaching and research farm. Samples of dairy cattle (36), beef cattle (26), sheep (19), horses (21), pigs (23), goats (23) and humans (13) were collected and screened for the presence of MRS. The detection of mecA gene was performed by PCR to determine the resistance of the samples to methicillin. Antimicrobial-resistance testing to penicillin, meropenem, ceftriaxone, cephalothin, oxacillin, levofloxacin, enrofloxacin, chloramphenicol, ciprofloxacin, gentamicin, clindamycin, erytromycin, linezolid, sulfamethoxazole/trimethoprim, tetracycline, doxycycline and vancomycin was performed on the mecA+ isolates. From the 161 samples, four methicillin-resistant coagulase-negative Staphylococci (MRCoNS) were isolated from human beings (31%), whereas none was isolated from animals (0%). No methicillin-resistant Staphylococcus aureus (MRSA) were isolated. All of the MRCoNS isolates from this work presented different antimicrobial resistance patterns. MRCoNS may be present in humans associated with animals while not present in the animals. Selective pressure outside of the farm and a lack of MRCoNS transmission between humans and animals may be responsible for this lack of correlation. © 2011 Blackwell Verlag GmbH.
Gómez, Paula; Lozano, Carmen; Benito, Daniel; Estepa, Vanesa; Tenorio, Carmen; Zarazaga, Myriam; Torres, Carmen
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
Watters Amy A
Full Text Available Abstract Background Daptomycin is a cyclic lipopeptide with potent activity and broad spectrum against Gram-positive bacteria currently used for the treatment of complicated skin and skin structure infections and bacteremia, including right sided endocarditis. We evaluated the in vitro activity of this compound and selected comparator agents tested against clinical strains of staphylococci and enterococci collected in European medical centers in 2005. Methods A total of 4,640 strains from 23 medical centers located in 10 European countries, Turkey and Israel (SENTRY Program platform were tested for susceptibility by reference broth microdilution methods according to Clinical and Laboratory Standards Institute guidelines and interpretative criteria. Mueller-Hinton broth was supplemented to 50 mg/L Ca++ for testing daptomycin. Results for oxacillin (methicillin-resistant staphylococci and vancomycin-resistant enterococci were analyzed separately. Results Oxacillin resistance rates among Staphylococcus aureus varied from 2.1% in Sweden to 42.5% in the United Kingdom (UK and 54.7% in Ireland (29.1% overall, while vancomycin resistance rates varied from 0.0% in France, Sweden and Switzerland to 66.7% in the UK and 71.4% in Ireland among Enterococcus faecium (17.9% overall. All S. aureus strains were inhibited at daptomycin MIC of 1 mg/L (MIC50/90, 0.25/0.5 mg/L; 100.0% susceptible and only one coagulase-negative staphylococci strain (0.1% showed an elevated (>1 mg/L daptomycin MIC value (4 mg/L. Among E. faecalis (MIC50/90, 0.5/1 mg/L; 100% susceptible the highest daptomycin MIC value was 2 mg/L; while among E. faecium (MIC50/90, 2/4 mg/L; 100% susceptible the highest MIC result was 4 mg/L. Conclusion Daptomycin showed excellent in vitro activity against staphylococci and enterococci collected in European medical centers in 2005 and resistance to oxacillin, vancomycin or quinupristin/dalfopristin did not compromise its activity overall against these
Shi, Guang-sen; Boost, Maureen; Cho, Pauline
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.
Maria Teresa Mascellino
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.
Vivian de Lima Spode Coutinho
Full Text Available INTRODUCTION: Resistance to macrolides, lincosamides and streptogramins B (MLS B antibiotics in staphylococci may be due to modification in ribosomal target methylase encoded by erm genes. The expression of MLS B resistance lead to three phenotypes, namely constitutive resistance (cMLS B, inducible resistance (iMLS B, and resistance only to macrolides and streptogramins B (MS B. The iMLS B resistance is the most difficult to detect in the clinical laboratory. OBJECTIVE: This study investigated the expression of MLS B resistance and the prevalence of the erm genes among 152 clinical isolates of Staphylococcus aureus and coagulase-negative Staphylococcus (CNS from Hospital de Clínicas de Porto Alegre. METHODS: Primary MLS B resistance was detected by the disk diffusion method. Isolates with iMLS B phenotype were tested by double-disk induction method. All isolates were tested by a genotypic assay, PCR with specific primers. RESULTS: A total of 46.7% of staphylococci were positive for cMLS B; 3.3% for iMLS B and 3.3% for MS B. One or more erm genes were present in 50.1% of isolates. The gene ermA was detected in 49 isolates, ermC in 29 and ermB in 3. CONCLUSION: The prevalence of the ermA, ermB and ermC genes were 29.6%, 17.1% and 0.66% respectively, and constitutive resistance was the most frequent as compared to the other two phenotypes.
negative staphylococci >(CoNS) and. Staphylococcus aureus that ... Keywords: vancomycin, methicillin resistance, Staphylococcus aureus, coagulase-negative staphylococci, antimicrobial agents ... freshly prepared 1% aqueous starch solution.
Patel, Nimesh; Miller, Darlene; Relhan, Nidhi; Flynn, Harry W
Rapid identification of pathogens causing endophthalmitis may improve treatment outcomes through early administration of species-specific medication. The current study reports a new molecular application of peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) with Staphylococcus-specific molecular PNA probes for the potential rapid detection of common pathogens causing endophthalmitis. An experimental study was designed to evaluate the proof of concept at the microbiology laboratory of the Bascom Palmer Eye Institute. Stored culture-positive staphylococci endophthalmitis isolates obtained from prior vitreous samples (n = 15), along with broth as negative controls (n = 5) were used. Inoculum was prepared to a final concentration of 1 × 105 colony-forming units/mL to ensure that the isolates were viable. Smears of samples were fixed and hybridized using QuickFISH protocol with probes for Staphylococcus. With PNA-FISH technique, Staphylococcus aureus was identified in 9 of 10 samples and coagulase-negative staphylococci were identified in 10 of 10 samples. Detection time was 20 minutes. This study serves a proof of concept using a new microbial detection system with FISH probes, and may have the potential for clinical use in the rapid and accurate identification of isolates from patients with endophthalmitis.
Species level identification of coagulase negative Staphylococcus spp. from buffalo using matrix-assisted laser desorption ionization-time of flight mass spectrometry and cydB real-time quantitative PCR.
Pizauro, Lucas J L; de Almeida, Camila C; Soltes, Glenn A; Slavic, Durda; Rossi-Junior, Oswaldo D; de Ávila, Fernando A; Zafalon, Luiz F; MacInnes, Janet I
Incorrect identification of Staphylococcus spp. can have serious clinical and zoonotic repercussions. Accordingly, the aim of this study was to determine if matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and/or cydB real- time quantitative PCR (qPCR) could be used to accurately identify coagulase negative Staphylococcus spp. (CoNS) obtained from buffalo milk and milking environment samples. Seventy-five of 84 CoNS isolates could be identified to the species level (score value >1.99) using MALDI-TOF MS. However, as determined by cytochrome d ubiquinol oxidase subunit II (cydB) qPCR and by 16S RNA and cydB gene sequencing, 10S. agnetis strains were wrongly identified as S. hyicus by MALDI-TOF MS. In addition, 9 isolates identified by MALDI-TOF only to the genus level (score values between 1.70 and 1.99) could be identified to species by cydB qPCR. Our findings suggest that MALDI-TOF MS is a reliable method for rapid identification of S. chromogenes and S. epidermidis (species of interest both in human and veterinary medicine) and may be able to correctly identify other Staphylococcus spp. However, at present not all Staphylococcus spp. found in buffalo milk can be accurately identified by MALDI-TOF MS and for these organisms, the cydB qPCR developed in the current study may provide a reliable alternative method for rapid identification of CoNS species. Copyright © 2017 Elsevier B.V. All rights reserved.
Cloxacillin versus vancomycin for presumed late-onset sepsis in the Neonatal Intensive Care Unit and the impact upon outcome of coagulase negative staphylococcal bacteremia: a retrospective cohort study.
Lawrence, Sarah L; Roth, Virginia; Slinger, Robert; Toye, Baldwin; Gaboury, Isabelle; Lemyre, Brigitte
Coagulase negative staphylococcus (CONS) is the main cause of late-onset sepsis in Neonatal Intensive Care Units (NICU). Although CONS rarely causes fulminant sepsis, vancomycin is frequently used as empiric therapy. Indiscriminate use of vancomycin has been linked to the emergence of vancomycin resistant organisms. The objective of this study was to compare duration of CONS sepsis and mortality before and after implementation of a policy of selective vancomycin use and compare use of vancomycin between the 2 time periods. A retrospective study was conducted of infants > or =4 days old, experiencing signs of sepsis with a first positive blood culture for CONS, during two 12-month periods. Late-onset sepsis was treated empirically with vancomycin and gentamicin during period 1, and cloxacillin and gentamicin during period 2. The confidence interval method was used to assess non-inferiority of the outcomes between the two study groups. There were 45 episodes of CONS sepsis during period 1 and 37 during period 2. Duration of sepsis was similar between periods (hazard ratio of 1.00, 95%CI: 0.64, 1.57). One death during period 2 was possibly related to CONS sepsis versus none in period 1. Vancomycin was used in 97.8% of episodes in period 1 versus 81.1% of episodes in period 2. Although we failed to show non-inferiority of duration of sepsis in the cloxacillin and gentamicin group compared to the vancomycin and gentamicin group, duration of sepsis was clinically similar. Restricting vancomycin for confirmed cases of CONS sepsis resistant to oxacillin appears effective and safe, and significantly reduces vancomycin use in the NICU.
Cloxacillin versus vancomycin for presumed late-onset sepsis in the Neonatal Intensive Care Unit and the impact upon outcome of coagulase negative staphylococcal bacteremia: a retrospective cohort study
Full Text Available Abstract Background Coagulase negative staphylococcus (CONS is the main cause of late-onset sepsis in Neonatal Intensive Care Units (NICU. Although CONS rarely causes fulminant sepsis, vancomycin is frequently used as empiric therapy. Indiscriminate use of vancomycin has been linked to the emergence of vancomycin resistant organisms. The objective of this study was to compare duration of CONS sepsis and mortality before and after implementation of a policy of selective vancomycin use and compare use of vancomycin between the 2 time periods. Methods A retrospective study was conducted of infants ≥4 days old, experiencing signs of sepsis with a first positive blood culture for CONS, during two 12-month periods. Late-onset sepsis was treated empirically with vancomycin and gentamicin during period 1, and cloxacillin and gentamicin during period 2. The confidence interval method was used to assess non-inferiority of the outcomes between the two study groups. Results There were 45 episodes of CONS sepsis during period 1 and 37 during period 2. Duration of sepsis was similar between periods (hazard ratio of 1.00, 95%CI: 0.64, 1.57. One death during period 2 was possibly related to CONS sepsis versus none in period 1. Vancomycin was used in 97.8% of episodes in period 1 versus 81.1% of episodes in period 2. Conclusion Although we failed to show non-inferiority of duration of sepsis in the cloxacillin and gentamicin group compared to the vancomycin and gentamicin group, duration of sepsis was clinically similar. Restricting vancomycin for confirmed cases of CONS sepsis resistant to oxacillin appears effective and safe, and significantly reduces vancomycin use in the NICU.
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
... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Coagulase plasma. 866.2160 Section 866.2160 Food... DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2160 Coagulase plasma. (a) Identification. Coagulase plasma is a device that consists of freeze-dried animal or human plasma that is...
Podkowik, Magdalena; Bystroń, Jarosław; Bania, Jacek
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.
Full Text Available PSM-mec is a secreted virulence factor that belongs to the phenol-soluble modulin (PSM family of amphipathic, alpha-helical peptide toxins produced by Staphylococcus species. All known PSMs are core genome-encoded with the exception of PSM-mec, whose gene is found in specific sub-types of SCCmec methicillin resistance mobile genetic elements present in methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci. In addition to the cytolytic translational product, PSM-mec, the psm-mec locus encodes a regulatory RNA. In S. aureus, the psm-mec locus influences cytolytic capacity, methicillin resistance, biofilm formation, cell spreading and the expression of other virulence factors, such as other PSMs, which results in a significant impact on immune evasion and disease. However, these effects are highly strain-dependent, which is possibly due to differences in PSM-mec peptide versus psm-mec RNA-controlled effects. Here, we summarize the functional properties of PSM-mec and the psm-mec RNA molecule and their roles in staphylococcal pathogenesis and physiology.
Cybulska, J.; Jeljaszewicz, J.
The growth of 100 staphylococcal strains in normal rabbit serum was studied. Determinations were carried out by the tube method previously described and by a new plate method, and the results agreed in both cases. The strains of staphylococci used differed in the production of bound coagulase, clumping factor, and staphylokinase, and exhibited different phage patterns. Their multiplication in serum was not dependent on any of the above-mentioned characteristics. The newly elaborated plate test is characterized by easy and quick performance and allows for the determination of 25 to 36 strains on one Petri plate. Images PMID:5320886
Ünal, Nilgün; Askar, Şinasi; Macun, Hasan Ceyhun; Sakarya, Fatma; Altun, Belgin; Yıldırım, Murat
The aims of this study were to determine the existence of pvl gene, some toxin genes, and mecA gene in Staphylococcus aureus strains isolated from sheep milk and to examine antimicrobial resistance profiles in staphylococci from sheep and goats' milk. The milk samples were collected from 13 different small ruminant farms in Kirikkale province from February to August 2009. A total of 1,604 half-udder milk samples from 857 ewes and 66 half-udder milk samples from 33 goats were collected. Staphylococcus spp. were isolated and identified from the samples. Toxin genes and mecA gene among S. aureus strains were determined by PCR. Antimicrobial susceptibility of staphylococci was examined by the disk diffusion method on Mueller-Hinton agar, and interpreted according to the Clinical Laboratory Standards Institute (CLSI) guidelines. The prevalence of subclinical intramammary infection in both ewes and goats was 5.2%. The most prevalent subclinical mastitis agents were coagulase-negative staphylococci and S. aureus with prevalences 2.8% (n:46) and 1.3% (n = 21), respectively. The prevalence of resistances in isolated Staphylococcus spp. to penicilin G, tetracycline, erythromycin, gentamicin, and enrofloxacin were found as 26.9% (18), 7.5% (5), 6.0% (4), 3.0% (2), and 1.5% (1), respectively. Only 3 of the 21 S. aureus ewe isolates (13.4%) were shown to harbor enterotoxin genes being either seh, sej or sec. However, fourteen (66.6%) of the 21 S. aureus isolates had pvl gene while none of the isolates harbored mecA gene. In conclusion, Staphylococci were shown to be the most prevalent bacteria isolated from subclinical mastitis of ewes and goats and these isolates were susceptible to most of the antibiotics. In addition, S. aureus strains isolated from ewes were harboring few staphylococcal enterotoxin genes. However, Panton-Valentine leukocidin produced by S. aureus could be an important virulence factor and contribute to subclinical mastitis pathogenicity.
Clifford C. Ugwu
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.
David R Cue
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.
Development of a new pentaplex real-time PCR assay for the identification of poly-microbial specimens containing Staphylococcus aureus and other staphylococci, with simultaneous detection of staphylococcal virulence and methicillin resistance markers.
Okolie, Charles E; Wooldridge, Karl G; Turner, David P; Cockayne, Alan; James, Richard
Staphylococcus aureus strains harbouring genes encoding virulence and antibiotic resistance are of public health importance. In clinical samples, pathogenic S. aureus is often mixed with putatively less pathogenic coagulase-negative staphylococci (CoNS), both of which can harbour mecA, the gene encoding staphylococcal methicillin-resistance. There have been previous attempts at distinguishing MRSA from MRCoNS, most of which were based on the detection of one of the pathognomonic markers of S. aureus, such as coa, nuc or spa. That approach might suffice for discrete colonies and mono-microbial samples; it is inadequate for identification of clinical specimens containing mixtures of S. aureus and CoNS. In the present study, a real-time pentaplex PCR assay has been developed which simultaneously detects markers for bacteria (16S rRNA), coagulase-negative staphylococcus (cns), S. aureus (spa), Panton-Valentine leukocidin (pvl) and methicillin resistance (mecA). Staphylococcal and non-staphylococcal bacterial strains (n = 283) were used to validate the new assay. The applicability of this test to clinical samples was evaluated using spiked blood cultures (n = 43) containing S. aureus and CoNS in mono-microbial and poly-microbial models, which showed that the 5 markers were all detected as expected. Cycling completes within 1 h, delivering 100% specificity, NPV and PPV with a detection limit of 1.0 × 10(1) to 3.0 × 10(1) colony forming units (CFU)/ml, suggesting direct applicability in routine diagnostic microbiology. This is the most multiplexed real-time PCR-based PVL-MRSA assay and the first detection of a unique marker for CoNS without recourse to the conventional elimination approach. There was no evidence that this new assay produced invalid/indeterminate test results. Copyright © 2015 Elsevier Ltd. All rights reserved.
María del Pilar Sánchez Bonilla
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.
Wang, Hye-Young; Kim, Sunghyun; Kim, Jungho; Park, Soon-Deok; Uh, Young; Lee, Hyeyoung
Methicillin-resistant Staphylococcus aureus (MRSA) is the most prevalent cause of bloodstream infections (BSIs) and is recognized as a major nosocomial pathogen. This study aimed to evaluate a newly designed multiplex real-time PCR assay capable of the simultaneous detection of mecA, S. aureus, and coagulase-negative staphylococci (CoNS) in blood culture specimens. The Real-MRSA and Real-MRCoNS multiplex real-time PCR assays (M&D, Republic of Korea) use the TaqMan probes 16S rRNA for Staphylococcus spp., the nuc gene for S. aureus, and the mecA gene for methicillin resistance. The detection limit of the multiplex real-time PCR assay was 10(3) CFU/ml per PCR for each gene target. The multiplex real-time PCR assay was evaluated using 118 clinical isolates from various specimen types and a total of 350 positive blood cultures from a continuous monitoring blood culture system. The results obtained with the multiplex real-time PCR assay for the three targets were in agreement with those of conventional identification and susceptibility testing methods except for one organism. Of 350 positive bottle cultures, the sensitivities of the multiplex real-time PCR kit were 100% (166/166 cultures), 97.2% (35/36 cultures), and 99.2% (117/118 cultures) for the 16S rRNA, nuc, and mecA genes, respectively, and the specificities for all three targets were 100%. The Real-MRSA and Real-MRCoNS multiplex real-time PCR assays are very useful for the rapid accurate diagnosis of staphylococcal BSIs. In addition, the Real-MRSA and Real-MRCoNS multiplex real-time PCR assays could have an important impact on the choice of appropriate antimicrobial therapy, based on detection of the mecA gene. Copyright © 2014, American Society for Microbiology. All Rights Reserved.
Wang, Hye-young; Kim, Sunghyun; Kim, Jungho; Park, Soon-Deok
Methicillin-resistant Staphylococcus aureus (MRSA) is the most prevalent cause of bloodstream infections (BSIs) and is recognized as a major nosocomial pathogen. This study aimed to evaluate a newly designed multiplex real-time PCR assay capable of the simultaneous detection of mecA, S. aureus, and coagulase-negative staphylococci (CoNS) in blood culture specimens. The Real-MRSA and Real-MRCoNS multiplex real-time PCR assays (M&D, Republic of Korea) use the TaqMan probes 16S rRNA for Staphylococcus spp., the nuc gene for S. aureus, and the mecA gene for methicillin resistance. The detection limit of the multiplex real-time PCR assay was 103 CFU/ml per PCR for each gene target. The multiplex real-time PCR assay was evaluated using 118 clinical isolates from various specimen types and a total of 350 positive blood cultures from a continuous monitoring blood culture system. The results obtained with the multiplex real-time PCR assay for the three targets were in agreement with those of conventional identification and susceptibility testing methods except for one organism. Of 350 positive bottle cultures, the sensitivities of the multiplex real-time PCR kit were 100% (166/166 cultures), 97.2% (35/36 cultures), and 99.2% (117/118 cultures) for the 16S rRNA, nuc, and mecA genes, respectively, and the specificities for all three targets were 100%. The Real-MRSA and Real-MRCoNS multiplex real-time PCR assays are very useful for the rapid accurate diagnosis of staphylococcal BSIs. In addition, the Real-MRSA and Real-MRCoNS multiplex real-time PCR assays could have an important impact on the choice of appropriate antimicrobial therapy, based on detection of the mecA gene. PMID:24648566
Ramos, Roberta Juliano; Pereira, Murilo Anderson; Miotto, Letícia Adélia; Faria, Renata D'Aquino; Silveira Junior, Nelson; Vieira, Cleide Rosana Werneck
The aim of this study was to assess the contamination of oysters (Crassostrea gigas), harvested in six different regions of the South Bay of Santa Catarina Island, with Coliforms at 45 ºC, Escherichia coli, Vibrio spp., positive coagulase staphylococci, and Salmonella sp. over a period of one year. One hundred eighty oyster samples were collected directly from their culture sites and analyzed. Each sample consisted of a pool of 12 oysters. All of the samples analyzed showed absence of Salmone...
Full Text Available Accurate and rapid typing of S. aureus is crucial to the control of its infections and minimizing its leakage to the food chain. The primary purpose of this research was to isolate S. aureus from camels’ meat and nasal swabs and to characterize the isolates for coagulase production and the presence of methicillin gene using PCR-RFLP of coagulase gene. A total of 264 camel’s meat and nasal swabs were collected from abattoirs or meat markets and were used in the study. Ninety two percent of samples showed typical colonies of S. aureus on Baird-Parker agar with a mean count 2.5 × 104 ± 1.8 × 104 CFU g-1. Upon confirmation of the isolates using S. aureus specific thermonuclease gene (nuc PCR primers, only 64 isolates contained the specific product and thus were confirmed as S. aureus. However, when tested for the presence of coagulase gene, only 48 of them were positive while the other 16 were coagulase negative. Coagulase gene-RFLP revealed 19 distinct patterns when the gene was digested with Alu I and Cfo I. The typing revealed that the 48 classified isolates were genetically diverse and comprised a heterogeneous population with 14 genotypes at a 44.4% similarity level. When the coagulase positive isolates were tested for the presence of methicillin resistance (mec A gene, 37 of the isolates were positive while the other 11 isolates were negative. The high heterogeneity among S. aureus isolates might be due to cross contamination between camel carcasses in slaughter houses and from handlers and their utensils.
Bernardi, Adilson A. C.; Pizzolitto, Elisabeth Loshchagin [UNESP; Pizzolitto, Antonio Carlos [UNESP
Slime production is an important virulence factor of coagulase-negative Staphylococcus spp., allowing them to attach to smooth surfaces of biomaterials, and it has been associated with infections of implanted medical devices. In the present study the production of slime capsules in 27 strains of coagulase-negative Staphylococcus was investigated by culture in Congo Red agar (77.7% positivity), spectrophotometric or microplate method (81.4% positivity) and scanning electron microscopy (88.9% p...
Turner, G. C.; Cox, P. E.
Cloxacillin-resistant strains of Staphylococcus aureus were detected by their ability to grow on agar containing 1·6 μg./ml. of cloxacillin, a more reliable method than the disc-diffusion test. At Sefton General Hospital, Liverpool, between 1963 and 1965, there was an increase in the number of infections among in-patients that were caused by staphylococci resistant to cloxacillin despite the fact that use of the antibiotic was largely confined to an isolation ward for patients with staphylococcal sepsis. Although there is no evidence yet that staphylococci resistant to cloxacillin will become as common in hospital practice as those resistant to penicillin and tetracycline it is clear that there is a need for continued vigilance and measures to prevent spread of staphylococci from infected patients. PMID:5188891
Baptiste, Keith Edward; Williams, Kerry; Williams, Nicola J.
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...
Objective: To determine the prevalence of antibiotic susceptibility of Staphylococcus species isolated from foremilk samples. Setting: Milk was collected from five farms within a 70 km radius of Gaborone, Botswana. Subjects: Two hundred and twenty five staphylococci isolates from foremilk samples. Main outcome measures: ...
carcasses ( saprophytic = growing on dead tissues). Sellin et al. (1975) at England’s University of Bristol Medical ., -School conducted a prospective...Barr, H. 1980. Urinary tract infection caused by Staphylococcus saprophyticus. J. Infect. Dis., 142:510-515. Kaye, D. 1976. Infecting microorganisms , p
BSI) due to their immune-compromised status, repeated hospitalizations, and various procedures. Staphylococci, particularly coagulase negative Staphylococci (CoNS) are noteworthy pathogens in such cases, and the emergence of.
Bendahou, Abdrezzak; Abid, Mohammed; Bouteldoun, Nadine; Catelejine, Dierick; Lebbadi, Mariam
The aim of this research was to determine the prevalence of enterotoxin genes (sea-seo) in Coagulase Positive Staphylococcus (CPS) isolated from unpasteurized milk and milk products. These results were compared with the results obtained by using the detection kit SET-RPLA for the specific detection of staphylococcal enterotoxins (SEA-SED). Eighty-one samples of milk and milk products were analyzed for the presence of Staphylococcus strains. Forty-six coagulase positive Staphylococcus isolates were tested for the production of staphylococcal enterotoxins (SEA-SED) by using the reversed passive latex agglutination method. The strains were also tested for the presence of se genes (sea-seo) by polymerase chain reaction. One or more classical enterotoxin products (SEA-SED) were observed in 39% of the strains tested, while se genes were detected in 56.5%. SEA and sea were most commonly detected. For newly discovered se genes among CPS isolates tested in this study, except the seh gene which was revealed in four isolates (8.7 %), none of the strains harbored any of the other se genes (see, seg, sei, sej, sek, sel, sem, seo and sen). The finding of a pathogen such as staphylococci-producing SEs and containing se genes in milk and milk products in northern Morocco may indicate a problem for public health in this region. The presence of enterotoxigenic strains in food does not always necessarily mean that the toxin will be produced. For that reason, the combination of both methods (RPLA and PCR) is a guarantee for success in diagnostic analysis tests.
Subramanian, Arunagiri; Chitalia, Vidushi K; Bangera, Kushminda; Vaidya, Shashikant P; Warke, Rajas; Chowdhary, Abhay; Deshmukh, Ranjana A
Staphylococcus aureus is a facultative anaerobic Gram positive coccal bacterium whose incidence ranges to different infections. It is a cause of various uncomplicated skin infections, abscesses, septicaemia/bacteraemia, gastroenteritis, endocarditis, toxic shock syndrome and food intoxications. Various methods with varied time, sensitivities, specificities and costs are available, but may not be used as a reliable test for the identification and differentiation of S. aureus. Therefore, there is a need to evaluate newer tests. To compare the conventional tests with a commercial available kit for reliable, cost effective identification and confirmation of S. aureus. The current prospective study was conducted in the Department of Clinical Pathology, Haffkine Institute for a period of six months. A total of 341 clinical isolates of staphylococci isolated from pus, urine, blood culture and sterile body fluids were subjected to conventional tests like Tube Coagulase Test (TCT) using Rabbit Plasma (RP) and Human Plasma (HP), culture media such as Mannitol Salt Agar (MSA) and Deoxyribonuclease (DNase) media in parallel to HiaureusTM Coagulase Confirmation Kit (HACCK), a commercially available kit for identification of S. aureus. Amplification of the femA gene was used as a comparative reference point test to calculate the sensitivity, specificity and concordance values of the conventional tests. Amongst the coagulase based tests, HACCK was 100% sensitive and specific. The TCT using RP was 98.58% sensitive while TCT using HP was less sensitive (95.37%). A total of 100% specificity was observed for TCT using RP while TCT using HP was 96.68% specific. The MSA and DNase media were 97.86% vs 96.44% and 96.67% vs 91.67% sensitive and specific respectively. The combination tests had varying sensitivity and specificity ranges. The HACCK demonstrated 100% concordance with femA amplification and was labelled as an ideal perfect test (κ=1) with MSA as an alternative test for S
Faria, Nuno A; Conceição, Teresa; Miragaia, Maria
. A total of 54 patients (80%) were colonized with staphylococci that included nine different species identified by internal transcribed spacer PCR (ITS-PCR) and 16S RNA sequencing. The highest rates of colonization were found for S. epidermidis (58%) and S. aureus (39%). Methicillin resistance was present......%). Carriage of multiple species was detected in 20 patients (30%), 16 of whom were colonized with both S. aureus and S. epidermidis. In two cases, simultaneous carriage of different methicillin resistant species was detected. However, the strains carried different SCCmec types. Additional studies in the same...
Anderson, J C
During the course of an acute, fatal mastitis in mice caused by intra-mammary injection of approximately 10(5) cfu of Staphylococcus aureus, small numbers of staphylococci were detected in liver, spleen, kidney and heart. Transport from the mammary gland was by the venous blood system with no evidence of lymph node involvement. Intravenous injection of 100 times more staphylococci (approximately 10(7) cfu) than were fatal by the intramammary route failed to kill the mice and small numbers of staphylococci were found in the visceral organs. However, intravenous injection of approximately 10(8) cfu of Staph. aureus was fatal and large numbers of staphylococci were found in the visceral organs. Treatment with cloxacillin (500 micrograms) by the intramammary or subcutaneous routes, or equally divided between these two routes, cleared the visceral organs of staphylococci; subcutaneous therapy had a limited and variable effect on numbers of staphylococci in the mammary gland while intramammary and divided treatments significantly reduced the numbers of staphylococci. Thus, staphylococci are disseminated to visceral organs during acute staphylococcal mastitis, but this makes a negligible contribution to the clinical disease.
Jaén, N; Martínez-Pastor, J C; Muñoz-Mahamud, E; García-Ramiro, S; Bosch, J; Mensa, J; Soriano, A
To update the clinical information of the 47 patients with a prosthetic joint infection due to Gram-negative bacilli included in a previous study and to reassess the predictors of failure after a longer follow-up. Using the electronic files of our hospital, all the information regarding readmissions to the hospital, new surgical procedures and the reason for the new surgery (infection, aseptic loosening), and the last visit in the hospital were registered. The medical chart of the 35 patients that were considered in remission in the previous publication was reviewed. In 30 patients no clinical evidence of failure was detected and no additional surgery on the previously infected prosthesis was necessary and they were considered in long-term remission. In 5 cases a late complication was identified. One case had a reinfection due to coagulase-negative staphylococci after 22 months from the open debridement and required a 2-stage revision surgery. The other 4 cases developed an aseptic loosening and it was necessary to perform a 1-stage exchange. Receiving a fluoroquinolone when all the Gram-negatives involved in the infection were susceptible to fluoroquinolones was the only factor associated with remission in the univariate analysis (p=0.002). After a long-term follow-up, our results support the importance of using fluoroquinolones in acute PJI due to Gram-negative bacilli.
Comparação entre o sistema Petrifilm RSA® e a metodologia convencional para a enumeração de estafilococos coagulase positiva em alimentos Comparison of Petrifilm RSA system with the traditional methodology for the enumeration of coagulase-positive Staphylococcus in foods
Anderson de Souza Sant'ana
Full Text Available A pesquisa de estafilococos coagulase positiva em alimentos é feita através de sua detecção e enumeração em meios seletivos e diferenciais e posterior caracterização pelos testes de coagulase, termonuclease, Gram e catalase. Estes testes podem requerer até quatro dias para obtenção dos resultados finais, além do tempo e material dispensados ao seu uso. As placas Petrfilm® RSA são uma alternativa a esta metodologia. No presente estudo, foram analisadas 62 amostras de diferentes alimentos comparando-se a metodologia tradicional (ABP, seguido dos testes de coagulaseou Staphytect Plus e o sistema Petrifilm. O sistema Petrifilm® RSA diferiu significativamente da metodologia tradicional, dando médias de contagem superiores. Ao se considerar colônias típicas e atípicas, o ABP seguido de confirmação pelo Staphytect Plus diferiu significativamente dos demais métodos testados. O Petrifilm RSA é uma alternativa para a enumeração de estafilococos coagulase positivos em alimentos, em virtude do menor tempo (aproximadamente 31 horas, para obter-se resultados realmente quantitativos. Deve-se testar tanto colônias típicas quanto atípicas crescidas no ABP, para se evitar falhas na quantificação dos resultados. Esta contagem é mais significativa ao se usar o teste de aglutinação em látex (Staphytect Plus. A análise de estafilococos em alimentos apresenta diversas limitações.Foods are examined for the presence of coagulase-positive staphylococci using selective and differential media followed by characterization for coagulase, thermoestable nuclease, Gram and catalase tests. These tests may require up to four days to obtain the results, besides time and material used. The Petrifilm® rapid S.aureus count plate is a alternative to traditional methodology. In this study, 62 samples of several foods were analysed comparing the BPA method followed by coagulase or latex agglutination tests and Petrifilm® RSA method. The Petrifilm
Oliveira, Ana Maria; Padovani, Carlos Roberto; Miya, Norma Teruko Nagô; Sant'ana, Anderson S; Pereira, José Luiz
In this study, the enterotoxigenic potential of Staphylococcus strains (n = 574) isolated from raw milk samples (n = 140) was determined for their capacity to produce staphylococcal enterotoxins. In addition, the relationship between the presence of enterotoxins, coagulase, and thermonuclease (Tnase) was assessed. The results showed that 19% of Staphylococcus was enterotoxigenic, being able to produce at least one of the staphylococcal enterotoxins (A, B, C, and D). Most of the strains were able to produce enterotoxin D (68.8%), whereas 12.8% of the Staphylococcus strains were able to produce staphylococcal enterotoxin A. Besides, the production of more than one type of enterotoxins by the same strain was observed. Tnase was considered the best marker for enterotoxigenic potential of isolates, although some of them were negative for coagulase and Tnase but positive for enterotoxin production. Therefore, either the use of Tnase to assess Staphylococcus enterotoxigenic potential or the use of simple and easy screening tests for enterotoxin production should receive more attention when evaluating the pathogenic potential of foodborne Staphylococcus strains. Due to the association of both coagulase positive Staphylococcus and coagulase negative Staphylococcus with foodborne disease outbreaks, regulators and industries should pay more attention to enterotoxigenic Staphylococcus rather than focusing only on S. aureus or coagulase positive Staphylococcus. Finally, data found here suggest a high risk of staphylococcal intoxication with the consumption of raw milk or dairy products made from raw milk.
, 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 ...
Vanderlinde, P B; Fegan, N; Mills, L; Desmarchelier, P M
Coagulase positive staphylococci isolated at an Australian abattoir, from beef carcasses, hands, and environmental samples, were typed by the DNA macrorestriction patterns obtained following PFGE of SmaI digests. The predominant PFGE pattern of isolates collected before evisceration was different from the dominant pattern of isolates collected after evisceration. PFGE patterns for isolates from workers' hands and from concordant carcasses were indistinguishable. PFGE types among isolates collected from non-evisceration abattoir personnel and clerical staff were distinct from patterns among isolates collected from the slaughter floor personnel. Twenty-nine (85%) of the 34 isolates collected from carcasses after evisceration exhibited PFGE patterns that were 93% homologous. During evisceration the carcasses were handled extensively and it is proposed that the hands of workers at this abattoir are the primary source of staphylococcal contamination of carcasses on the slaughter floor. This paper highlights the usefulness of PFGE as an epidemiological tool for determining the source of staphylococcal contamination in the food industry.
Ziad Jaradat; Akram Al Aboudi; Mahmoud Shatnawi; Qotaibah Ababneh
.... The primary purpose of this research was to isolate S. aureus from camels' meat and nasal swabs and to characterize the isolates for coagulase production and the presence of methicillin gene using PCR-RFLP of coagulase gene...
Desmarchelier, P M; Higgs, G M; Mills, L; Sullivan, A M; Vanderlinde, P B
The contamination of beef carcasses with coagulase-positive staphylococci (CPS) was studied at three beef abattoirs (A, B and C). The incidence and the number of CPS were determined on cattle hides immediately after slaughter and on three carcass sites (brisket, flank and round) at different points during processing along the slaughter line. The incidence of CPS on cattle hides ranged from 20 to 68.6%. At abattoir A, 6.5% of the carcasses sampled before evisceration were contaminated with CPS, compared to 40% of the carcasses after evisceration. The incidence on carcasses changed little during further processing; however, after chilling for 72 h, the incidence increased to 83%. After evisceration, the brisket and flank areas were more often contaminated than the round. A similar pattern of contamination was observed at abattoir B. At abattoir C, 26.7% of the samples collected before evisceration were contaminated and this fell to 16.7% after evisceration. After chilling for 72 h, the incidence of carcass contamination with CPS increased to 46.7%. The average number of CPS on contaminated carcasses prior to and after overnight chilling was less than 50 colony-forming units (cfu)/cm2 and, after weekend chilling, increased to 64 and 112 cfu/cm2 in abattoirs A and B, respectively. Of the isolates tested, 71.4% produced staphylococcal enterotoxin and 21% could not be classified phenotypically. The hands of workers and environmental sites associated with the evisceration process were examined for CPS at abattoir A. Hands were heavily contaminated and were the likely source of CPS contamination at this abattoir.
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.
Beça, Nuno; Bessa, Lucinda Janete; Mendes, Ângelo; Santos, Joana; Leite-Martins, Liliana; Matos, Augusto J F; da Costa, Paulo Martins
Staphylococcus pseudintermedius is the most prevalent coagulase-positive Staphylococcus inhabitant of the skin and mucosa of dogs and cats, causing skin and soft tissue infections in these animals. In this study, coagulase-positive Staphylococcus species were isolated from companion animals, veterinary professionals, and objects from a clinical veterinary environment by using two particular culture media, Baird-Parker RPF agar and CHROMagar Staph aureus. Different morphology features of colonies on the media allowed the identification of the species, which was confirmed by performing a multiplex polymerase chain reaction (PCR). Among 23 animals, 15 (65.2%) harbored coagulase-positive Staphylococcus, being 12 Staphylococcus pseudintermedius carriers. Four out of 12 were methicillin-resistant S. pseudintermedius (MRSP). All veterinary professionals had coagulase-positive Staphylococcus (CoPS) species on their hands and two out of nine objects sampled harbored MRSP. The antimicrobial-resistance pattern was achieved for all isolates, revealing the presence of many multidrug-resistant CoPS, particularly S. pseudintermedius . The combined analysis of the antimicrobial-resistance patterns shown by the isolates led to the hypothesis that there is a possible crosscontamination and dissemination of S. aureus and S. pseudintermedius species between the three types of carriers sampled in this study that could facilitate the spread of the methicillin-resistance phenotype.
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.
Agarwal, Astha; Jain, Amita
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. 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. 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. 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.
Sansevere, Emily A; Robinson, D Ashley
Horizontal gene transfer plays a significant role in spreading antimicrobial resistance and virulence genes throughout the genus Staphylococcus , which includes species of clinical relevance to humans and animals. While phages and plasmids are the most well-studied agents of horizontal gene transfer in staphylococci, the contribution of integrative conjugative elements (ICEs) has been mostly overlooked. Experimental work demonstrating the activity of ICEs in staphylococci remained frozen for years after initial work in the 1980s that showed Tn 916 was capable of transfer from Enterococcus to Staphylococcus . However, recent work has begun to thaw this field. To date, 2 families of ICEs have been identified among staphylococci - Tn 916 that includes the Tn 5801 subfamily, and ICE 6013 that includes at least 7 subfamilies. Both Tn 5801 and ICE 6013 commonly occur in clinical strains of S. aureus . Tn 5801 is the most studied of the Tn 916 family elements in staphylococci and encodes tetracycline resistance and a protein that, when expressed in Escherichia coli , inhibits restriction barriers to incoming DNA. ICE 6013 is among the shortest known ICEs, but it still includes many uncharacterized open reading frames. This element uses an IS 30 -like transposase as its recombinase, providing some versatility in integration sites. ICE 6013 also conjugatively transfers among receptive S. aureus strains at relatively higher frequency than Tn 5801 . Continued study of these mobile genetic elements may reveal the full extent to which ICEs impact horizontal gene transfer and the evolution of staphylococci.
María Espino Hernández
unit of Hospital Ginecobstétrico "América Arias" in Havana, Cuba, from 1998 to 2000. METHODS: A retrospective study was carried out of the clinical and laboratory results obtained in 15 patients with sepsis caused by multiresistant staphylococci who received combined therapy with azlocillin and amikacin, according to hospital guidelines on the use of antibiotics. We used a broth microdilution method to study the patterns of resistance shown by isolated strains to 10 of the antibiotics in use. In vitro synergy tests, specifically the checkerboard technique with microtitration plates, were used to observe the effects of treatment in 8 patients. RESULTS: Twelve coagulase-negative staphylococci and three Staphylococcus aureus isolates showed five different patterns of resistance on the basis of their sensitivity to oxacillin, three aminoglycosides, and vancomycin. Six of the synergy tests showed a considerable synergistic effect, with an average three-fold reduction in the minimum inhibitory concentrations (MIC of the two antibiotics used to treat the patients. No antagonistic effects were noted, and the combined antibiotics showed an overall clinical effectiveness of 91.7%. CONCLUSIONS: The test showed that the therapeutic combination used was effective, but further studies are needed before conclusive results are obtained.
Noble, W. C.; Lloyd, D. H.; Appiah, S. N.
In an acute model of skin infection with Dermatophilus congolensis in the mouse, lesions can be prevented by simultaneous application of staphylococci which produce antibiotics; non-producer staphylococci fail to inhibit lesion formation.
Noble, W C; Lloyd, D H; Appiah, S N
In an acute model of skin infection with Dermatophilus congolensis in the mouse, lesions can be prevented by simultaneous application of staphylococci which produce antibiotics; non-producer staphylococci fail to inhibit lesion formation.
Camila Alencar Moreira; Lis Christina de Oliveira; Marina Silveira Mendes; Thiago de Melo Santiago; Eduardo Bedê Barros; Cibele Barreto Mano de Carvalho
This study determined the species of 54 staphylococci isolates from canine otitis and their ability to produce biofilm through the Congo red agar method, confirmed by scanning electron microscopy. The most frequently identified species were S. intermedius and S. simulans. Results showed that 30% of the strains were biofilm producers.
Camila Alencar Moreira
Full Text Available This study determined the species of 54 staphylococci isolates from canine otitis and their ability to produce biofilm through the Congo red agar method, confirmed by scanning electron microscopy. The most frequently identified species were S. intermedius and S. simulans. Results showed that 30% of the strains were biofilm producers.
Moreira, Camila Alencar; de Oliveira, Lis Christina; Mendes, Marina Silveira; Santiago, Thiago de Melo; Barros, Eduardo Bedê; de Carvalho, Cibele Barreto Mano
This study determined the species of 54 staphylococci isolates from canine otitis and their ability to produce biofilm through the Congo red agar method, confirmed by scanning electron microscopy. The most frequently identified species were S. intermedius and S. simulans. Results showed that 30% of the strains were biofilm producers.
Abstract. 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 ...
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 ...
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 ...
Bunke, C M; Brier, M E; Golper, T A
The use of the "peritonitis rate" in the management of patients undergoing peritoneal dialysis is assuming importance in comparing the prowess of facilities, care givers and new innovations. For this to be a meaningful outcome measure, the type of infection (causative pathogen) must have less clinical significance than the number of infections during a time interval. The natural history of Staphylococcus aureus, pseudomonas, and fungal peritonitis would not support that the outcome of an episode of peritonitis is independent of the causative pathogen. Could this concern be extended to other more frequently occurring pathogens? To address this, the Network 9 Peritonitis Study identified 530 episodes of single organism peritonitis caused by a gram positive organism and 136 episodes caused by a single non-pseudomonal gram negative (NPGN) pathogen. Coincidental soft tissue infections (exit site or tunnel) occurred equally in both groups. Outcomes of peritonitis were analyzed by organism classification and by presence or absence of a soft tissue infection. NPGN peritonitis was associated with significantly more frequent catheter loss, hospitalization, and technique failure and was less likely to resolve regardless of the presence or absence of a soft tissue infection. Hospitalization and death tended to occur more frequently with enterococcal peritonitis than with other gram positive peritonitis. The outcomes in the NPGN peritonitis group were significantly worse (resolution, catheter loss, hospitalization, technique failure) compared to coagulase negative staphylococcal or S. aureus peritonitis, regardless of the presence or absence of a coincidental soft tissue infection. Furthermore, for the first time, the poor outcomes of gram negative peritonitis are shown to be independent of pseudomonas or polymicrobial involvement or soft tissue infections. The gram negative organism appears to be the important factor. In addition, the outcome of peritonitis caused by S. aureus
Aarestrup, Frank Møller; Dangler, C. A.; Sordillo, L. M.
for restriction fragment length polymorphism (RFLP) of the coagulase gene. A variable region of the coagulase gene was amplified using the polymerase chain reaction (PCR) followed by AluI restriction enzyme digestion. A total of 15 different RFLP patterns were observed. The predominant pattern was found in 35...
Hettinga, K.A.; Valenberg, van H.J.F.; Lam, T.J.G.M.; Hooijdonk, van A.C.M.
The ability to detect mastitis pathogens based on their volatile metabolites was studied. Milk samples from cows with clinical mastitis, caused by Staphylococcus aureus, coagulase-negative staphylococci, Streptococcus uberis, Streptococcus dysgalactiae, and Escherichia coli were collected. In
The isolates in 66 (64.71%) cases were Gram positive of which 36 (54.55%) were Streptococcus spp., 24 (36.36%) Staphylococcus aureus and 6 (9.09%) cases coagulase negative Staphylococcus (CONS). Both Streptococci and coagulase negative Staphylococci were highly sensitive (100%) to Linezolid, Vancomycin and ...
Walther, Birgit; Hermes, Julia; Cuny, Christiane; Wieler, Lothar H; Vincze, Szilvia; Abou Elnaga, Yassmin; Stamm, Ivonne; Kopp, Peter A; Kohn, Barbara; Witte, Wolfgang; Jansen, Andreas; Conraths, Franz J; Semmler, Torsten; Eckmanns, Tim; Lübke-Becker, Antina
.... CPS- nasal carriage, different aspects of human-to-dog relationship as well as potential interspecies transmission risk factors were investigated by offering nasal swabs and a questionnaire to dog owners (108) and their dogs (108...
Staphylococci constituted 30.7% of the total bacterial isolates with Staphylococcus aureus accounting for only 10.2%. Other Gram-positive cocci cultured include, Staphylococcus spp (20.5%), coagulase negative staphylococci (CONS) (12.8%) and Streptococcus spp (1.2%). Gram-negative rods constituted 55.1% of the total ...
Meyer, Cornelia; Fredriksson-Ahomaa, Maria; Stüber, Elisabeth; Thiel, Susanne; Märtlbauer, Erwin
Staphylococcus aureus are a hazard to human health since they can cause infections and food poisoning. Antimicrobial resistant strains render the treatment of infections problematic and contribute to the spread of antimicrobial resistance. They are therefore of great public concern. This study determined the resistance pattern of coagulase-positive S. aureus (CPSA) isolated from nasal swabs of 100 slaughter pigs from one farm in Uruguay. Out of 69 animals, 71 CPSA were collected. Minimum inhibitory concentrations of 20 antimicrobials were determined using the broth microdilution method in accordance with CLSI recommendations. No methicillin-resistant S. aureus were detected. All CPSA were resistant to three or more classes of antimicrobials (i.e., multiresistant), whereby all CPSA were resistant to spectinomycin. Most of the isolates (46%) were resistant to six classes of antimicrobials. Almost all isolates were resistant to penicillin (99%), ampicillin (99%), gentamicin (96%), tetracycline (90%), and tilmicosin (87%). Very high resistance rates were observed against erythromycin (77%) and clindamycin (70%). High resistance was observed against tiamulin (40%), enrofloxacin (31%), and florfenicol (23%) and low resistance was observed against amoxicillin/clavulanic acid (4%). All CPSA isolates were mecA negative. The results of the present study could be related to an overuse of antimicrobials in pig production and should encourage veterinarians and pig holders to practice a controlled administration of chemotherapeutics in pig husbandry.
Robert D. Wojtyczka
Full Text Available Synergistic interactions between commonly used antibiotics and natural bioactive compounds may exhibit therapeutic benefits in a clinical setting. Berberine, an isoquinoline-type alkaloid isolated from many kinds of medicinal plants, has proven efficacy against a broad spectrum of microorganisms. The aim of the presented work was to assess the antibacterial activity of berberine chloride in light of the effect exerted by common antibiotics on fourteen reference strains of Staphylococccus spp., and to evaluate the magnitude of interactions of berberine with these antistaphylococcal antibiotics. In our study minimum inhibitory concentrations (MIC of berberine chloride against CoNS ranged from 16 to 512 µg/mL. The most noticeable effects were observed for S. haemolyticus ATCC 29970, S. epidermidis ATCC 12228, S. capitis subsp. capitis ATCC 35661, S. galinarium ATCC 700401, S. hominis subsp. hominis ATCC 27844, S. intermedius ATCC 29663 and S. lugdunensis ATCC 49576. The most significant synergistic effect was noticed for berberine in combination with linezolid, cefoxitin and erythromycin. The synergy between berberine and antibiotics demonstrates the potential application of compound combinations as an efficient, novel therapeutic tool for antibiotic-resistant bacterial infections.
Ian R Monk
Full Text Available Most strains of Staphylococcus aureus and Staphylococcus epidermidis possess a strong restriction barrier that hinders exchange of DNA. Recently, major advances have been made in identifying and characterizing the restriction-modification systems involved. In particular a novel type IV restriction enzyme that recognizes cytosine methylated DNA has been shown to be the major barrier to transfer of plasmid DNA from Escherichia coli into S. aureus and S. epidermidis. One or sometimes two type I restriction-modification systems provide an additional barrier. Here we review the recent advances in understanding of restriction systems in staphylococci and highlight how this has been exploited to improve our ability to manipulate genetically previously untransformable strains.
ABSTRACT: This study identified some virulence factors in staphylococci isolated from the faecal samples of children in Ile-. Ife, Nigeria and also determined the relationships in the expressions of these virulent factors in the S. aureus and coagulase negative ... phenotypic identification of staphylococci was performed by ...
Tirosh-Levy, Sharon; Steinman, Amir; Carmeli, Yehuda; Klement, Eyal; Navon-Venezia, Shiri
Methicillin-resistant staphylococci (MRS), and specifically Methicillin-resistant Staphylococcus aureus (MRSA) colonization or infection have become a serious emerging condition in equine hospitals, with complex concerns regarding animals, personnel and public health. The objectives of this study were to evaluate the prevalence and risk factors for colonization by Staphylococci, MRS, and MRSA among horses in Israel. Nasal swabs were collected from horses at 17 riding stables (n=206), and from hospitalized horses admitted to a veterinary hospital (n=84). Species identification was performed by pta gene PCR, RFLP analysis and sequencing. MRS was identified by the presence of mecA. Genetic relatedness of MRSA isolates was determined by spa typing and MLST. SCCmec-type and pvl gene were determined. Univariable and multivariable statistical analysis were used to identify potential risk factors. Colonization with Staphylococci was found among 3.8% of farm horses and 50.6% of hospitalized horses (pMRSA was found exclusively among hospitalized horses (7.2%). All MRSA isolates belonged to a unique single multi-drug-resistant clone, ST5-SCCmec V, pvl-negative, spa-type t535. Risk factors for colonization with MRS were pure bred, hospitalization and antibiotic use. This is the first surveillance study of Staphylococci in horses in Israel, and the first report on the presence of a unique MRSA strain among hospital horses, recognizing the veterinary hospital as a potential reservoir for MRSA, an antibiotic resistant pathogen with human relevance. Copyright © 2015. Published by Elsevier B.V.
Wilkins, Judd R.; Hatten, Mary B.; Salmirs, Seymour
After horizontal rotation on a clinostat at 1 rev/min, subsurface colonies of staphylococci in soft agar were compact and spherical; nonrotated colonies were diffuse and elongated. Images PMID:5369302
Kateete, David P; Kimani, Cyrus N; Katabazi, Fred A; Okeng, Alfred; Okee, Moses S; Nanteza, Ann; Joloba, Moses L; Najjuka, Florence C
The ideal identification of Staphylococcus aureus clinical isolates requires a battery of tests and this is costly in resource limited settings. In many developing countries, the tube coagulase test is usually confirmatory for S. aureus and is routinely done using either human or sheep plasma. This study evaluated Mannitol salt agar and the deoxyribonuclease (DNase) test for improving the efficiency of the tube coagulase test in resource limited settings. The efficiency of human and sheep plasma with tube coagulase tests was also evaluated. One hundred and eighty Gram positive, Catalase positive cocci occurring in pairs, short chains or clusters were subjected to growth on Mannitol salt agar, deoxyribonuclease and tube coagulase tests. Of these, isolates that were positive for at least two of the three tests (n = 60) were used to evaluate the performance of the tube coagulase test for identification of S. aureus, using PCR-amplification of the nuc gene as a gold standard. Human plasma was more sensitive than sheep plasma for the tube coagulase test (sensitivity of 91% vs. 81% respectively), but both plasmas had very low specificity (11% and 7% respectively). The sensitivity and specificity of the tube coagulase test (human plasma) was markedly improved when Mannitol salt agar and DNase were introduced as a tri-combination test for routine identification of Staphylococcus aureus (100% specificity and 75% sensitivity). The specificity and sensitivity of Mannitol salt agar/DNase/tube coagulase (sheep plasma) combination was 100% and 67%, respectively. The efficiency of the tube coagulase test can be markedly improved by sequel testing of the isolates with Mannitol salt agar, DNase and Tube coagulase. There is no single phenotypic test (including tube coagulase) that can guarantee reliable results in the identification of Staphylococcus aureus.
Kateete David P
Full Text Available Abstract Background The ideal identification of Staphylococcus aureus clinical isolates requires a battery of tests and this is costly in resource limited settings. In many developing countries, the tube coagulase test is usually confirmatory for S. aureus and is routinely done using either human or sheep plasma. This study evaluated Mannitol salt agar and the deoxyribonuclease (DNase test for improving the efficiency of the tube coagulase test in resource limited settings. The efficiency of human and sheep plasma with tube coagulase tests was also evaluated. Methods One hundred and eighty Gram positive, Catalase positive cocci occurring in pairs, short chains or clusters were subjected to growth on Mannitol salt agar, deoxyribonuclease and tube coagulase tests. Of these, isolates that were positive for at least two of the three tests (n = 60 were used to evaluate the performance of the tube coagulase test for identification of S. aureus, using PCR-amplification of the nuc gene as a gold standard. Results Human plasma was more sensitive than sheep plasma for the tube coagulase test (sensitivity of 91% vs. 81% respectively, but both plasmas had very low specificity (11% and 7% respectively. The sensitivity and specificity of the tube coagulase test (human plasma was markedly improved when Mannitol salt agar and DNase were introduced as a tri-combination test for routine identification of Staphylococcus aureus (100% specificity and 75% sensitivity. The specificity and sensitivity of Mannitol salt agar/DNase/tube coagulase (sheep plasma combination was 100% and 67%, respectively. Conclusion The efficiency of the tube coagulase test can be markedly improved by sequel testing of the isolates with Mannitol salt agar, DNase and Tube coagulase. There is no single phenotypic test (including tube coagulase that can guarantee reliable results in the identification of Staphylococcus aureus.
Bina Pani Gupta
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.
Schwab, Frank; Gastmeier, Petra; Meyer, Elisabeth
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
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
Ziad Jaradat; Akram Al Aboudi; Mahmoud Shatnawi; Qotaibah Ababneh
Accurate and rapid typing of S. aureus is crucial to the control of its infections and minimizing its leakage to the food chain. The primary purpose of this research was to isolate S. aureus from camels’ meat and nasal swabs and to characterize the isolates for coagulase production and the presence of methicillin gene using PCR-RFLP of coagulase gene. A total of 264 camel’s meat and nasal swabs were collected from abattoirs or meat markets and were used in the study. Ninety two percent of sam...
Skirrow, M. B.; Rogers, K. B.
Two hundred and thirty-seven selected, penicillin-resistant isolates of Staph. aureus collected over a 13-month period were examined retrospectively for cloxacillin/methicillin resistance; 44 were found to be resistant. Six other strains which had been preserved from an earlier period were also found to be resistant. All 50 cloxacillin-resistant strains were in addition resistant to streptomycin and several other antibiotics. According to their resistance patterns and reactions to phages, they could be classified into two categories. The proportion of cloxacillin-resistant strains among the total number of staphylococci isolated during the 13-month period was estimated to be 2·7%. Children most at risk from infection were those undergoing long-term hospital treatment and on several occasions children transferred from other hospitals in the region were found to be carrying resistant strains. Most infections caused by these strains were minor but they were nevertheless capable of producing severe illness and they were responsible for 33% of all staphylococcal wound infections. PMID:5206516
Darwish, Samah F.; Asfour, Hanaa A. E.
A total of 40 S. aureus and 68 coagulase negative Staphylococcus (CNS) isolates from bovine subclinical mastitis were investigated for their ability to form biofilm as one of the most important virulence factors.Using Congo Red Agar (CRA) method, 32.5%, 35%, and 32.5% of S. aureus strains were strong, intermediate, and negative biofilm producers, while in CNS the percentages were 29.5%, 42.6%, and 27.9%, respectively. By microtiter plate (MTP) method, 52.5%, 27.5%, and 20% of S. aureus isolates were strong, moderate, and weak biofilm producers, while in CNS the percentages were 44%, 30.9%, and 19.2%, respectively. Indian ink staining was used to detect the EPS layer of biofilm producers. All isolates were screened for presence of biofilm related genes, eno, icaA, icaD, and bap. In S. aureus isolates, the positive rates of eno, icaA, icaD, and bap genes were 75%, 15%, 62.5%, and 2.5% while in CNS were 92.6%, 5.9%, 47.1%, and 4.4%, respectively. The eno gene had the highest rate while the bap gene had the lowest rate. Presence of icaA and icaD genes was not always correlated with biofilm production. This study demonstrated high prevalence of Staphylococcus biofilm producers among bovine mastitis in Egypt. Therefore, attention must be paid toward implementation of new ways for effective treatment of such infections. PMID:24298212
Stevens, M; Piepers, S; Supré, K; De Vliegher, S
The main objectives of this study were to quantify the consumption of antimicrobials on a convenience sample of dairy herds and to determine the association between herd-level antimicrobial consumption and inhibition zone diameters (IZD) of non-aureus staphylococci and Staphylococcus aureus isolates from subclinical mastitis cases. Also, the association between the IZD of non-aureus staphylococci and Staph. aureus isolates within a herd was studied. Antimicrobial consumption data on 56 Flemish dairy farms were obtained between 2013 and 2014 by so-called garbage can audits and expressed as antimicrobial treatment incidence (ATI), with the unit of ATI being the number of defined daily doses animal (DDDA) used per 1,000 cow-days. The average total ATI in adult dairy cattle for all active substances was 18.73 DDDA per 1,000 cow-days and ranged from 6.28 to 42.13 DDDA between herds. The ATI of critically important (for human health) antimicrobials was 6.91 DDDA per 1,000 cow-days; that is, 37% of total antimicrobial consumption. The average ATI for intramammary therapy of (sub)clinical mastitis, intramammary dry-cow therapy, and systemically administered therapy was 5.20, 6.70, and 6.73 DDDA, respectively. The IZD of 239 non-aureus staphylococci and 88 Staph. aureus isolates originating from milk samples from cows with subclinical mastitis collected on selected dairy herds were determined using Kirby-Bauer disk diffusion and ranged between 6 and 42 mm. Because only a limited number of clinical breakpoints (Clinical and Laboratory Standards Institute) and epidemiological cut-off values (European Committee on Antimicrobial Susceptibility Testing) are available for mastitis-causing bacteria in bovine, IZD were used as a proxy for antimicrobial resistance. Inhibition zone diameters of non-aureus staphylococci for cefquinome, a critically important β-lactam antibiotic, were negatively associated with the ATI of critically important β-lactam for systemically administered
Nowakiewicz, Aneta; Ziółkowska, Grażyna; Zięba, Przemysław; Gnat, Sebastian; Wojtanowicz-Markiewicz, Katarzyna; Trościańczyk, Aleksandra
The aim of the study was molecular analysis of coagulase-positive isolates of Staphylococcus bacteria obtained from wild animals and evaluation of their resistance to antimicrobial agents. A total of 76 rectal swabs were taken from wild animals. The species of the Staphylococcus isolates was determined by MALDI TOF MS, susceptibility to antimicrobials was evaluated by phenotypic and molecular methods, epidemiological analysis (ADSRRS-fingerprinting) was also carried out. MRSA isolate was typed by MLST and spa-typing. The animals tested, were carriers (n=38) of coagulase-positive Staphylococcus (S. aureus, S. pseudintermedius and S. delphini B). Analyzed isolates were resistant to 1 or 2 antimicrobials, which was confirmed by the presence of genes (blaZ, ermA, ermB, msrA, tetK and tetM). A multi-drug resistant and methicillin-resistant isolate of S. aureus was obtained as well (MRSA, ST8, t1635, PVL-positive and ACME-negative). The ADSRRS-fingerprinting method enabled interspecific and intraspecific differentiation of coagulase-positive Staphylococcus isolates, revealing a certain degree of correlation between the species of the isolate, and the degree of similarity between the isolates. The presence of resistance genes in 13% (5/38) of the isolates obtained from wild animals, including one methicillin-resistant isolate, is relatively small in comparison to the degree of colonization by resistant strains in humans, livestock or pets. Nevertheless, due to the possibility of contact between wild animals, domestic animals and humans, transmission of resistant strains is possible, as suggested by our isolation of a MRSA strain typed as ST8 and specific spa type t1635, which had previously been isolated exclusively from humans. Copyright © 2015 Elsevier Ltd. All rights reserved.
Sampimon, O.C.; Vernooij, J.C.A.; Mevius, D.J.; Sol, J.
De laatste jaren worden in Nederland in toenemende mate coagulase negatieve staphylococcen geïsoleerd uit melkmonsters van melkkoeien. Van de bacteriologisch positieve melkmonsters, afkomstig van koeien met een subklinische mastitis, werden bij GD in 1999 in 16,2 procent en in 2004 in 42,2 procent
Yamashita, Kenji; Shimizu, Akira; Kawano, Junichi; Uchida, Eiji; Haruna, Akihiro; Igimi, Shizunobu
Staphylococci were isolated from the external auditory meatus in 14 (48.3%) of 29 dogs affected with otitis externa (OE dogs) and 28 (68.3%) of 41 dogs without OE (non-OE dogs). Twenty-two OE isolates were identified as belonging to 12 species, and 42 non-OE isolates were identified as belonging to 13 species. The predominant species found in both OE and non-OE isolates were S. intermedius, and S. epidermidis. Thirty-eight (59.4%) of 64 isolates were resistant to one or more of the 17 antimicrobial agents tested. Resistance to PCG and ABPC was most frequent. S. schleiferi subsp. coagulans, a recent etiologic agent of canine OE, was isolated from OE and non-OE dogs. All of the 5 S. schleiferi subsp. coagulans isolates showed typical characteristics. No clear difference in the extracellular enzyme or toxin profiles, nor in the PFGE patterns, was demonstrated between the OE and non-OE isolates of S. schleiferi subsp. coagulans. A new PCR primer set specific for 16S rDNA was designed to identify strains of S. schleiferi subsp. coagulans. The amplified fragment was detected in all of the 5 isolates as well as the type strain GA 211 (=JCM 7470) and a reference strain GA 11, but was not detected in any strains of the related species, S. aureus, S. intermedius and S. hyicus. The PCR may allow a simple, rapid and precise identification of S. schleiferi subsp. coagulans, in addition to the standard tube test for free coagulase.
Full Text Available 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.
Etinosa O. Igbinosa
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.
Marcela Vyletělová; Oto Hanuš; Renáta Karpíšková; Zora Šťástková
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...
Groves, David J.; Short, H.; Thewaini, A. J.; Young, Frank E.
Staphylococci were isolated from rural and urban populations in Iraq, which were not known to be exposed to either heavy metals or antibiotics. The antibiotic and heavy metal resistance patterns of these strains were analyzed in both mannitol-fermenting and nonfermenting strains. Over 90% of the strains were resistant to at least one of the following antibiotics: penicillin, chloramphenicol, erythromycin, tetracycline, cephalothin, lincomycin, or methicillin. In general, mannitol-fermenting strains were resistant to penicillin and cupric ions. Mannitol-negative strains were more frequently associated with mercuric ion and tetracycline resistance. Although resistance to penicillin and tetracycline can coexist, the combination of penicillin resistance and tetracycline resistance usually occurred in mannitol-negative strains. The possibility of selection of heavy metal-resistant strains due to exposure to toxic levels of methylmercury was examined. No significant increase in mercuric ion-resistant strains of staphylococci or Escherichia coli were detected in exposed populations as compared to control groups. The possible reasons for this result are discussed. PMID:1147593
Carson, Domonique A; Barkema, Herman W; Naushad, Sohail; De Buck, Jeroen
Non-aureus staphylococci (NAS), the bacteria most commonly isolated from the bovine udder, potentially protect the udder against infection by major mastitis pathogens due to bacteriocin production. In this study, we determined the inhibitory capability of 441 bovine NAS isolates (comprising 26 species) against bovine Staphylococcus aureus Furthermore, inhibiting isolates were tested against a human methicillin-resistant S. aureus (MRSA) isolate using a cross-streaking method. We determined the presence of bacteriocin clusters in NAS whole genomes using genome mining tools, BLAST, and comparison of genomes of closely related inhibiting and noninhibiting isolates and determined the genetic organization of any identified bacteriocin biosynthetic gene clusters. Forty isolates from 9 species (S. capitis, S. chromogenes, S. epidermidis, S. pasteuri, S. saprophyticus, S. sciuri, S. simulans, S. warneri, and S. xylosus) inhibited growth of S. aureus in vitro, 23 isolates of which, from S. capitis, S. chromogenes, S. epidermidis, S. pasteuri, S. simulans, and S. xylosus, also inhibited MRSA. One hundred five putative bacteriocin gene clusters encompassing 6 different classes (lanthipeptides, sactipeptides, lasso peptides, class IIa, class IIc, and class IId) in 95 whole genomes from 16 species were identified. A total of 25 novel bacteriocin precursors were described. In conclusion, NAS from bovine mammary glands are a source of potential bacteriocins, with >21% being possible producers, representing potential for future characterization and prospective clinical applications.IMPORTANCE Mastitis (particularly infections caused by Staphylococcus aureus) costs Canadian dairy producers $400 million/year and is the leading cause of antibiotic use on dairy farms. With increasing antibiotic resistance and regulations regarding use, there is impetus to explore bacteriocins (bacterially produced antimicrobial peptides) for treatment and prevention of bacterial infections. We
Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis - doi: 10.4025/actascibiolsci.v32i4.6276 Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis - doi: 10.4025/actascibiolsci.v32i4.6276
Ulisses de Pádua Pereira
Full Text Available Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis. Three hundred fourty four strains of coagulase-positive Staphylococcus (CPS, isolated from mastitis cases, underwent phenotypic and genotypic tests to evaluate the efficiency of a simplified key, based on phenotypic tests for the discrimination of these microorganisms. The tests consisted of amplification of the femA gene and hemolysis in blood agar, production of acetoin and fermentation of maltose, mannitol and trehalose. Strains that showed negative results in the amplification test of the femA gene or that were not identified as Staphylococcus aureus (S. aureus by phenotypic tests were tested with the APISTAPH kit (Biomériux-France, for precise identification of species. Phenotypic tests revealed 338 strains (98.25% as S. aureus, three strains (0.86% as Staphylococcus hyicus, and three microorganisms (0.86% as Staphylococcus intermedius. PCR demonstrated that 338 (98.25% strains belonged to the S. aureus species, confirming the results for 336 strains from 338 identified, through a simplified phenotypic key. A high rate of correlation (98.83% was verifeid between the results of genotypic and phenotypic tests for the identification of S. aureus, demonstrating the applicability of the proposed key, for the discrimination of this microorganism in CPS isolated from bovine mastitis.Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis. Three hundred fourty four strains of coagulase-positive Staphylococcus (CPS, isolated from mastitis cases, underwent phenotypic and genotypic tests to evaluate the efficiency of a simplified key, based on phenotypic tests for the discrimination of these microorganisms. The tests consisted of amplification of the femA gene and hemolysis in blood agar, production of acetoin and fermentation of maltose, mannitol and trehalose
Terezinha Feitosa Machado; Maria de Fátima Borges; Bruna de Castro Porto; Cívita Teixeira de Sousa; Francisco Edílson Moreno de Oliveira
Níveis elevados de contaminantes microbiológicos, comumente encontrados em alimentos de origem animal e ambientes de processamento de alimentos, são aptos de impedir o crescimento de patógenos nestes produtos. Em alguns contextos onde bactérias ácido láticas (BAL) constituem a microbiota dominante, como nos produtos lácteos, Staphylococcus coagulase positiva (SCP) colonizam, persistem e produzem intoxicação alimentar. Com o objetivo de verificar a possível interferência da microbiota encontra...
Kateete, David P; Kimani, Cyrus N; Katabazi, Fred A; Okeng, Alfred; Okee, Moses S; Nanteza, Ann; Joloba, Moses L; Najjuka, Florence C
Abstract Background The ideal identification of Staphylococcus aureus clinical isolates requires a battery of tests and this is costly in resource limited settings. In many developing countries, the tube coagulase test is usually confirmatory for S. aureus and is routinely done using either human or sheep plasma. This study evaluated Mannitol salt agar and the deoxyribonuclease (DNase) test for improving the efficiency of the tube coagulase test in resource limited settings. The efficiency of...
Mohammad Mehdi Soltan Dallal
Full Text Available Coagulase is considered as a major determinant factor for the identification of Staphylococcus aureus strains. The 3′-end coding region of the coagulase (coa gene contains a series of 81-bp tandem repeats, which differ in the number and location of enzymatic restriction sites among different isolates. coa PCR-RFLP has been used widely to type S. aureus isolates in epidemiological studies. The current study was conducted to investigate the coagulase gene polymorphisms in S. aureus isolated from various food samples using an in house PCR-RFLP method. A total of 100 strains of S. aureus were isolated from food samples. Isolates were typed by PCR-RFLP analysis using NdeI restriction digestion of the coagulase gene PCR products. Results showed that amplification of coagulase genes from S. aureus produced different PCR products. The isolates were grouped into 18 genotypes using RFLP analysis results of the genes. In this study, the S. aureus isolates have been shown to include more than one coagulase genotype, but only had a few coa genotypes predominated.
Langlois, B E; Dawson, K.; Akers, K
Staphylococci which were considered to be lysostaphin susceptible on P agar containing Bacto-Agar showed different degrees of resistance to lysostaphin when tested on P agar made with BiTek agar. As a result, lysostaphin-susceptible strains were misidentified as lysostaphin-resistant strains.
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
Hend M. Abdulghany
Full Text Available The current study aimed to use Coagulase gene polymorphism to identify methicillin-resistant Staphylococcus aureus (MRSA subtypes isolated from nasal carriers in Minia governorate, Egypt, evaluate the efficiency of these methods in discriminating variable strains, and compare these subtypes with antibiotypes. A total of 400 specimens were collected from nasal carriers in Minia governorate, Egypt, between March 2012 and April 2013. Fifty-eight strains (14.5% were isolated and identified by standard microbiological methods as MRSA. The identified isolates were tested by Coagulase gene RFLP typing. Out of 58 MRSA isolates 15 coa types were classified, and the amplification products showed multiple bands (1, 2, 3, 4, 5, and 8 bands. Coagulase gene PCR-RFLPs exhibited 10 patterns that ranged from 1 to 8 fragments with AluI digestion. Antimicrobial susceptibility testing with a panel of 8 antimicrobial agents showed 6 different antibiotypes. Antibiotype 1 was the most common phenotype with 82.7%. The results have demonstrated that many new variants of the coa gene are present in Minia, Egypt, different from those reported in the previous studies. So surveillance of MRSA should be continued.
Full Text Available Background: Staphylococcus aureus is a Gram-positive, non-sporulated, lack of capsule, immovable, aerobic and anaerobic and able to tolerate high levels of salt (up to 15 percent. Therefore, foods containing salt provides an ideal environment for bacterial growth. It is also one of the most important bacterial born of spoilable food products, especially in dairy products. Lighvan traditional cheese is consumed in many regions of Iran, especially consumption of these cheeses is very high in Tehran which in the manufacture of such cheeses, the unpasteurized and raw milk is used and the sanitary conditions during processing and subsequent storage are low. So, considering the importance of this issue in the release of poisoning, the coagulase-positive Staphylococcus aureus contamination in Lighvan cheese on Retail Stores was evaluated. Materials and Methods: In order to perform this study, 22 samples of traditional Lighvan cheese were collected randomly from Tehran stores and according to Iran national standards and by using of bird-parker, Cook Meat cultures and coagulase test were evaluated. Results: Total of 22 samples, 14 samples (63/6% were contaminated by S. aureus over standard and 5 (22/7% were contaminated with Staphylococcus aureus coagulase positive. Conclusion: This study demonstrated that sanitary conditions should be observed during the production and Use of pasteurized milk in cheese production.
Riber, U; Espersen, F; Kharazmi, A
The ability to consume complement and activate neutrophils was investigated for staphylococci adherent to silicone surfaces and non-adherent staphylococci. Staphylococcus epidermidis strain ATCC 14990 and Staphylococcus aureus strain E 2371 were used in this study. The bacteria were allowed...... to adhere to silicone catheter segments for 2 h at 37 degrees C. Complement consumption was measured by reduction in serum haemolytic activity against sheep red blood cells. The induction of chemiluminescence was measured after opsonization of the staphylococci in 20% AB-positive human serum for 60 min...... with phagocytosis and elimination of S. epidermidis during the early establishment of a foreign body infection....
Suarez, María José; Arias, María Laura; del Mar Gamboa, María
Staphylococcus aureus is a pathogenic bacterium, widely distributed on nature and associated to general infection and food borne outbreaks. The relationship between this bacterium and food borne outbreaks has been done, historically, using several tests, including coagulase, thermonuclease and actually, PCR for the genes codifying for the enterotoxin responsible of clinical symptoms. The objective of this work is to detect enterotoxin A codifying gene through PCR in a group of S. aureus strains isolated from food samples, and also to correlate the presence of this gene with the production of coagulase and thermonuclease enzymes. A total of 69 staphylococcal strains were analyzed, 58 obtained from non pasteurized milk samples from the Estación Experimental Alfredo Volio Mata and 11 from the Food and Water Microbiology Laboratory collection, Universidad de Costa Rica. Coagulase, thermonuclease and enterotoxin A were analyzed in all the strains, and a statistical correlation was performed in order to verify possible associations. Results show that there is no correlation between the three variables, nevertheless, all coagulase positive strains were thermonuclease positive, and all enterotoxin positive strains were coagulase and thermonuclease positive, but not inversely. These results show that the use of presumptive or indirect tests for establishing entorotoxigenity of S. aureus strains is not truthful, more sensible and specific analysis, as PCR, shall be performed.
Full Text Available θ-Defensins are cyclic antimicrobial peptides expressed in leukocytes of Old world monkeys. To get insight into their antibacterial mode of action, we studied the activity of RTDs (rhesus macaque θ-defensins against staphylococci. We found that in contrast to other defensins, RTDs do not interfere with peptidoglycan biosynthesis, but rather induce bacterial lysis in staphylococci by interaction with the bacterial membrane and/or release of cell wall lytic enzymes. Potassium efflux experiments and membrane potential measurements revealed that the membrane impairment by RTDs strongly depends on the energization of the membrane. In addition, RTD treatment caused the release of Atl-derived cell wall lytic enzymes probably by interaction with membrane-bound lipoteichoic acid. Thus, the premature and uncontrolled activity of these enzymes contributes strongly to the overall killing by θ-defensins. Interestingly, a similar mode of action has been described for Pep5, an antimicrobial peptide of bacterial origin.
Zulfiqar A Mirani, Afsheen Aqeel, Shagufta Naz
Full Text Available Objectives: This study was designed to determine the prevalence of Staphylococci, Staphylococcus aureus (S. aureus, and Methicillin Resistant S. aureus (MRSA in commercially prepared food items that involve human handling, in Karachi-Pakistan. Methods: In a cross-sectional survey approach, in total 1012 food samples were analyzed. These samples were provided by different food processing industries during 2013 to 2016. Barid-Parker agar plates with egg yolk tellurite (Oxoid along with Mannitol Salt Agar (BioM, Staph-chromo agar (Merck, Staphylococcus 110 Agar (BioM, and Blood Agar (Oxoid were used for isolation and identification of Staphylococci. Polymerase chain reaction (PCR was used for amplification of mecA gene and specie identification via 16s RNA. Results: Total 723 samples (71.4% showed the presence of Staphylococci. Out of 723 staphylococcal isolates, 367 (36.2% were S. aureus and 85 (8.3% isolates were confirmed as MRSA. Molecular studies for MRSA typing showed that most of the isolates (74.1% belong to SCCmecA IV and 20% MRSA isolates were SCCmecA type II and 5.8% isolates carried SCCmecA type III. The MRSA isolates of SCCmecA type II belong to agr type I, while majority (67% of the isolates carry agr type II. Conclusion: This study suggested that majority of MRSA isolates recovered from commercial food items belongs to SCCmecA IV and human handling is a major factor for introduction of staphylococci in processed food items. J Microbiol Infect Dis 2017; 7(2: 83-87
Cardoso, Rosemari Laura; Maboni, Franciele; Machado, Gustavo; Alves, Sydney Hartz; de Vargas, Agueda Castagna
The aims of this study were to evaluate the antimicrobial potential of propolis extract by determining the minimum bactericidal concentration (MBC) for coagulase-positive Staphylococcus isolates (Staphylococcus aureus and Staphylococcus intermedius) and the minimum fungicidal concentration (MFC) for Malassezia pachydermatis isolates. The microorganisms were assayed using broth microdilution techniques. The MBC(90) was 21 mg mL(-1), and the MFC(90) was 5.3 mg mL(-1). The propolis extract was found to exhibit antimicrobial activity against both pathogens. Copyright 2009 Elsevier B.V. All rights reserved.
Dan, M; Marien, G J; Goldsand, G
Endocarditis caused by Staphylococcus warneri and necessitating valve replacement occurred in a previously healthy 32-year-old patient following vasectomy. No sign of an underlying valvular defect was noted during the operation. S. warneri is a recently identified species of coagulase-negative staphylococci. Endocarditis caused by coagulase-negative staphylococci is uncommon in young, healthy patients with normal heart valves and has not previously been described as a complication of vasectomy. Similarly, infections caused by S. warneri have not previously been described in humans.
Abdulaziz Ahmed Hashi
Full Text Available Staphylococcus pettenkoferi is a relatively recently described coagulase-negative staphylococci species first described in 2002. Since then, nine additional cases of infection caused by this species have been reported in various countries around the world, including Germany, Belgium, France, South Korea, Italy, Brazil and Mexico. The present report describes a case of S pettenkoferi peripheral line-associated bacteremia. To our knowledge, the present report is the first description of human infection caused by S pettenkoferi in Canada. The present report also provides an overview of the laboratory detection of uncommon coagulase-negative staphylococci.
Diversity of Staphylococcus Species Strains Based on Partial kat (Catalase) Gene Sequences and Design of a PCR-Restriction Fragment Length Polymorphism Assay for Identification and Differentiation of Coagulase-Positive Species (S. aureus, S. delphini, S. hyicus, S. intermedius, S. pseudintermedius, and S. schleiferi subsp. coagulans)▿
Blaiotta, Giuseppe; Fusco, Vincenzina; Ercolini, Danilo; Pepe, Olimpia; Coppola, Salvatore
A set of degenerate PCR primers was designed and used to amplify and sequence about 75% of the catalase (kat) gene from each of 49 staphylococcal strains. In some strains of Staphylococcus xylosus, S. saprophyticus, and S. equorum, two catalase genes, katA and katB, were found. A phylogenetic tree was generated and showed diversities among 66 partial (about 900-bp) staphylococcal kat nucleotide sequences (including 17 sequences found in GenBank) representing 26 different species. The topology of this tree showed a distribution of staphylococcal species similar, but not identical, to those reported previously based on 16S rRNA, hsp60, sodA, rpoB, tuf, and gap genes. The kat gene sequences were less conserved than those of 16S rRNA, rpoB, hsp60, and tuf genes and slightly more conserved than those of the gap gene. Therefore, kat gene sequence analysis may provide an additional marker for inferring phylogenetic relationships of staphylococci. Moreover, the discrete nucleotide polymorphism revealed in this gene could be exploited for rapid, low-cost identification of staphylococcal species through PCR-restriction fragment length polymorphism (RFLP) analysis. In this study, a PCR-RFLP assay performed by using only the TaqI restriction enzyme was successfully developed for rapid unequivocal identification/differentiation, at species and subspecies levels, of coagulase-positive staphylococci (CPS). The assay was validated by testing the DNA from 100 staphylococcal strains, including reference and wild CPS strains isolated from different environments. This reliable, rapid, and low-cost approach (requiring about 6 h from DNA isolation to the achievement of results and <5 Euros for each strain tested) allowed unambiguous identification of all the strains assayed, including the newly described S. delphini and S. pseudintermedius CPS species. PMID:19889901
Kamelia Mahmoud Osman; Jihan Badr; Khalid Al-Maary; Ihab Moussa; Ashgan Hessain; Zeinab Girh; Usama Abo-shama; Ahmed Orabi; Aalaa Saad
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 resista...
Coton, Emmanuel; Mulder, Niels; Coton, Monika; Pochet, Sylvie; Trip, Hein; Lolkema, Juke S.
A multiplex PCR method, aimed at the detection of genes associated with biogenic amine production, identified the odc gene encoding ornithine decarboxylase in 1 of 15 strains of Staphylococcus epidermidis. The ability of the positive strain, S. epidermidis 2015B, to produce putrescine in vitro was
Full Text Available Resumo: A mastite é a principal afecção do gado destinado à produção leiteira, que impacta significativamente a cadeia produtiva do leite, com reflexos ainda para a saúde pública. Estudou-se aspectos relacionados à etiologia, celularidade e de contagem bacteriana em 10 propriedades leiteiras, localizadas no Estado de São Paulo. Foram examinadas 1148 vacas em lactação, totalizando 4584 glândulas mamárias. Foram considerados os casos, em que houve isolamento de estafilococos coagulase positiva (SCP e estafilococos coagulase negativa (SCN. Os resultados revelaram microbiota com vários patógenos e diferentes espécies de SCN (128 casos e SCP (45, Staphylococcus aureus(90, Streptococcus agalactiae(70, Streptococcus dysgalactiae (69, Streptococcus uberis(29, Corynebacteriumspp. (230, Klebsiella pneumoniae(28, Klebsiella oxytoca(2, Escherichia coli(15, Enterobactersp. (3. Os resultados de contagem de células somáticas (CCS relacionados aos SCP e SCN não mostraram diferenças entre as propriedades avaliadas, entretanto com diferenças significantes ao se avaliar a CCS entre os dois grupos de estafilococos, como pode ser evidenciado ao comparar SCN Discreto e SCP exuberante (P<0,01, SCP Discreto e SCP exuberante (P<0,001 e SCN moderado e SCP exuberante (P<0,01. A avaliação da CCS relacionada à intensidade da infecção, considerando-se como crescimento discreto o isolamento de até nove colônias, moderado de dez a 29 colônias e exuberante, com 30 ou mais colônias, revelou para ambos os grupos de estafilococos que quanto maior o número de unidades formadoras de colônias (UFC, a CCS é mais elevada, sendo sempre maior nos casos de SCP. Conclui-se que quando há maior número de UFC, há concomitantemente maior CCS/mL de leite, no caso dos SCP e SCN, o que mostra relação direta da intensidade do processo infeccioso com a resposta da celularidade do leite, bem como pela relevância desses na etiologia das mastites e dos aspectos
Terezinha Feitosa Machado
Full Text Available Níveis elevados de contaminantes microbiológicos, comumente encontrados em alimentos de origem animal e ambientes de processamento de alimentos, são aptos de impedir o crescimento de patógenos nestes produtos. Em alguns contextos onde bactérias ácido láticas (BAL constituem a microbiota dominante, como nos produtos lácteos, Staphylococcus coagulase positiva (SCP colonizam, persistem e produzem intoxicação alimentar. Com o objetivo de verificar a possível interferência da microbiota encontrada no queijo Coalho sobre a presença de SCP, 64 amostras provenientes de 16 laticínios foram submetidas a análises microbiológicas para determinar os níveis de microrganismos aeróbios mesófilos (MAM, BAL e SCP. Os resultados obtidos mostraram que a microbiota autóctone das amostras analisadas não gerou condições inadequadas ao crescimento, desenvolvimento e isolamento de SCP, uma vez que este patógeno foi detectado mesmo nas amostras que apresentaram altos níveis de contagens de MAM e BAL.High levels of microbial contamination, commonly found in animal origin foods and food processing environments, are able to hinder the growth in these products. In some contexts where lactic acid bacteria (LAB are the normal dominant microbiota, such as in fermented food, Staphylococcus coagulase positive (SCP colonises, persists and produces food poisoning. With the aim of verifying the interference of autochthonous microbiota encountered in Curd cheese over the presence of Staphylococcus aureus, 64 samples from 16 dairy were submitted to microbiological analysis to determine the levels of Microorganisms aerobes mesophilics (MAM, LAB and SCP. The results showed that the indigenous microbiota of the samples did not lead to inadequate growth, development and isolation of SCP, since this pathogen was detected even in samples with high levels of counts of AM and BAL.
Mar 3, 2005 ... Escherichia Coli. Total gram negative. CONS. Enrerococci. Alkaligens Fecali. Staph. aureus. Strept. Viridan. GBS 1. Total gram positive. CONS= Coagulase negative staphylococci, GBS= Lancefield group B beta haemolytic streptococci. Cephalosporins (third generation) and clavulanic acid potentiated ...
de Mello, Jozi Fagundes; da Rocha, Laura Braga; Lopes, Ester Souza; Frazzon, Jeverson; da Costa, Marisa
Sanitary conditions are essential for the production of meals and control of the presence of pathogensis important to guarantee the health of customers. The aim of this study was to evaluate the sanitary quality of food services by checking the presence of thermotolerant coliforms, Staphylococcus sp. and evaluate the toxigenic potential from the latter. The analysis was performed on water, surfaces, equipment, ready-to-eat foods, hands and nasal cavity of handlers in seven food services. The water used in food services proved to be suitable for the production of meals. Most food, equipment and surfaces showed poor sanitary conditions due to the presence of thermotolerant coliforms (60.6%). Twenty-six Staphylococcus species were identified from the 121 Staphylococcus isolates tested. Staphylococci coagulase-negative species were predominant in the foods, equipment and surfaces. In food handlers and foods, the predominant species was Staphylococcus epidermidis. Twelve different genotypes were found after PCR for the classical enterotoxin genes. The seb gene (19.8%) was the most prevalent among all Staphylococcus sp. Both coagulase-positive and coagulase-negative Staphylococci showed some of the genes of the enterotoxins tested. We conclude that there are hygienic and sanitary deficiencies in the food services analyzed. Although coagulase-positive Staphylococci have not been present in foods there is a wide dispersion of enterotoxigenic coagulase-negative Staphylococci in the environment and in the foods analyzed, indicating a risk to consumer health.
Jozi Fagundes de Mello
Full Text Available Sanitary conditions are essential for the production of meals and control of the presence of pathogensis important to guarantee the health of customers. The aim of this study was to evaluate the sanitary quality of food services by checking the presence of thermotolerant coliforms, Staphylococcus sp. and evaluate the toxigenic potential from the latter. The analysis was performed on water, surfaces, equipment, ready-to-eat foods, hands and nasal cavity of handlers in seven food services. The water used in food services proved to be suitable for the production of meals. Most food, equipment and surfaces showed poor sanitary conditions due to the presence of thermotolerant coliforms (60.6%. Twenty-six Staphylococcus species were identified from the 121 Staphylococcus isolates tested. Staphylococci coagulase-negative species were predominant in the foods, equipment and surfaces. In food handlers and foods, the predominant species was Staphylococcus epidermidis. Twelve different genotypes were found after PCR for the classical enterotoxin genes. The seb gene (19.8% was the most prevalent among all Staphylococcus sp. Both coagulase-positive and coagulase-negative Staphylococci showed some of the genes of the enterotoxins tested. We conclude that there are hygienic and sanitary deficiencies in the food services analyzed. Although coagulase-positive Staphylococci have not been present in foods there is a wide dispersion of enterotoxigenic coagulase-negative Staphylococci in the environment and in the foods analyzed, indicating a risk to consumer health.
de Mello, Jozi Fagundes; da Rocha, Laura Braga; Lopes, Ester Souza; Frazzon, Jeverson; da Costa, Marisa
Sanitary conditions are essential for the production of meals and control of the presence of pathogensis important to guarantee the health of customers. The aim of this study was to evaluate the sanitary quality of food services by checking the presence of thermotolerant coliforms, Staphylococcus sp. and evaluate the toxigenic potential from the latter. The analysis was performed on water, surfaces, equipment, ready-to-eat foods, hands and nasal cavity of handlers in seven food services. The water used in food services proved to be suitable for the production of meals. Most food, equipment and surfaces showed poor sanitary conditions due to the presence of thermotolerant coliforms (60.6%). Twenty-six Staphylococcus species were identified from the 121 Staphylococcus isolates tested. Staphylococci coagulase-negative species were predominant in the foods, equipment and surfaces. In food handlers and foods, the predominant species was Staphylococcus epidermidis. Twelve different genotypes were found after PCR for the classical enterotoxin genes. The seb gene (19.8%) was the most prevalent among all Staphylococcus sp. Both coagulase-positive and coagulase-negative Staphylococci showed some of the genes of the enterotoxins tested. We conclude that there are hygienic and sanitary deficiencies in the food services analyzed. Although coagulase-positive Staphylococci have not been present in foods there is a wide dispersion of enterotoxigenic coagulase-negative Staphylococci in the environment and in the foods analyzed, indicating a risk to consumer health. PMID:25477940
Alessandra Siddi; Stefania Mannelli; Simona Roveta
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...
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. ...
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.
Sherman, Erica; Bayles, Kenneth; Moormeier, Derek; Wei, Timothy
Staphylococci bacteria are recognized as the most frequent cause of biofilm-associated infections. Although humans are regularly exposed to staphylococcus bacteria without consequence, a localized staph infection has the potential to enter the bloodstream and lead to serious infections such as endocarditis, pneumonia, or toxic shock syndrome. The mechanics of staphylococci biofilm formation and dispersion through the bloodstream are not well known. It has recently been observed that under certain flow conditions, bacteria grow in stable bio-films. Under other conditions, they organize in tower-like structures which break and are transported downstream by the flow. The fundamental questions addressed in this study are i) whether or not fluid mechanics plays a role in differentiating between film or tower formation and ii) whether or not the faulty towers are a bio-film propagation mechanism. This talk focuses on the application of micro-PIV to study this problem. Bacteria were cultured in a glass microchannel and subjected to a range of steady shear rates. Micro-PIV measurements were made to map the flow over and around different types of bio-film structures. Measurements and control volume analysis will be presented quantifying forces acting on these structures.
M. N. Boukhatem
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.
M. N. Boukhatem
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.
Lin, J L; Peng, Y; Ou, Q T; Lin, D X; Li, Y; Ye, X H; Zhou, J L; Yao, Z J
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
Third generation cephalosporin resistant Enterobacteriaceae and multidrug resistant gram-negative bacteria causing bacteremia in febrile neutropenia adult cancer patients in Lebanon, broad spectrum antibiotics use as a major risk factor, and correlation with poor prognosis.
Moghnieh, Rima; Estaitieh, Nour; Mugharbil, Anas; Jisr, Tamima; Abdallah, Dania I; Ziade, Fouad; Sinno, Loubna; Ibrahim, Ahmad
Bacteremia remains a major cause of life-threatening complications in patients receiving anticancer chemotherapy. The spectrum and susceptibility profiles of causative microorganisms differ with time and place. Data from Lebanon are scarce. We aim at evaluating the epidemiology of bacteremia in cancer patients in a university hospital in Lebanon, emphasizing antibiotic resistance and risk factors of multi-drug resistant organism (MDRO)-associated bacteremia. This is a retrospective study of 75 episodes of bacteremia occurring in febrile neutropenic patients admitted to the hematology-oncology unit at Makassed General Hospital, Lebanon, from October 2009-January 2012. It corresponds to epidemiological data on bacteremia episodes in febrile neutropenic cancer patients including antimicrobial resistance and identification of risk factors associated with third generation cephalosporin resistance (3GCR) and MDRO-associated bacteremia. Out of 75 bacteremias, 42.7% were gram-positive (GP), and 57.3% were gram-negative (GN). GP bacteremias were mostly due to methicillin-resistant coagulase negative staphylococci (28% of total bacteremias and 66% of GP bacteremias). Among the GN bacteremias, Escherichia coli (22.7% of total, 39.5% of GN organisms) and Klebsiella pneumoniae(13.3% of total, 23.3% of GN organisms) were the most important causative agents. GN bacteremia due to 3GC sensitive (3GCS) bacteria represented 28% of total bacteremias, while 29% were due to 3GCR bacteria and 9% were due to carbapenem-resistant organisms. There was a significant correlation between bacteremia with MDRO and subsequent intubation, sepsis and mortality. Among potential risk factors, only broad spectrum antibiotic intake >4 days before bacteremia was found to be statistically significant for acquisition of 3GCR bacteria. Using carbapenems or piperacillin/tazobactam>4 days before bacteremia was significantly associated with the emergence of MDRO (p < 0.05). Our findings have major
The main claim of this paper is that a general theory of negative concord (NC) should allow for the possibility of NC involving scoping of a universal quantifier above negation. I propose that Greek NC instantiates this option. Greek n-words will be analyzed as polarity sensitive universal
Colacito, A.; de Jongh, D.; Vargas Sandoval, A.L.
Minimal logic, i.e., intuitionistic logic without the ex falso principle, is investigated in its original form with a negation symbol instead of a symbol denoting the contradiction. A Kripke semantics is developed for minimal logic and its sublogics with a still weaker negation by introducing a
Negative Leadership by Colonel David M. Oberlander United States Army United States Army War...SUBTITLE Negative Leadership 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) Colonel David M...Dr. Richard C. Bullis Department of Command Leadership , and Management 8. PERFORMING ORGANIZATION REPORT NUMBER 9. SPONSORING/MONITORING
Hemels, Marieke A. C.; van den Hoogen, Agnes; Verboon-Maciolek, Malgorzata A.; Fleer, Andre; Krediet, Tannette G.
Objectives: Indwelling central venous catheters are the most important risk factors for the development of sepsis attributable to coagulase-negative staphylococci among preterm infants admitted to neonatal intensive care units. In addition, removal of a central venous catheter also may cause
Overall, 38% of the cultures yielded growth, while 52% of the nurses yielded growth as against 75% of the ward-maids. The bacterial isolates are Staphylococcus aureus (27%), Coagulase negative Staphylococci (20%), Bacillus alvei (17%), and Pseudomonas aeruginosa (4%). Out of the S. aureus isolates, 92.6% of them ...
Jiang, Saiping; Zheng, Beiwen; Ding, Wenchao; Lv, Longxian; Ji, Jinru; Zhang, Hua; Xiao, Yonghong; Li, Lanjuan
Staphylococcus hominis is a commensal coagulase-negative species of staphylococci. It has been considered a presumptive and opportunistic pathogen that causes nosocomial infections in humans. Here we present the draft genome sequence of S. hominis ZBW5, a multidrug-resistant strain isolated from a human skin sample, which provides opportunities to understand the mechanism and genetic basis of its pathogenesis.
Mar 3, 2005 ... Staph. aureus. Strept. Viridan. GBS 1. Total gram positive. CONS= Coagulase negative staphylococci, GBS= Lancefield group B beta haemolytic streptococci. Cephalosporins (third generation) and clavulanic acid potentiated amoxycillin offered the least rates of resistance while ampicillin had the highest ...
Sørensen, L P; Mark, T; Sørensen, M K
to â‚¬570 per mastitis case and were highest for contagious pathogens such as Staph. aureus and coagulase-negative staphylococci and lowest for Strep. dysgalactiae and Strep. uberis. The value for unspecific mastitis was â‚¬231 per case. Selection differentials (in â‚¬) were estimated for 4 different...
Jarløv, J O; Hansen, J E; Rosdahl, V T
A new typing method for Staphylococcus epidermidis was developed. Four biotinylated lectins--wheat germ agglutinin (WGA), soy bean agglutinin (SBA), lentil agglutinin (LCA) and Concanavalin A (ConA)--were added to immobilised whole cells of coagulase-negative staphylococci (CNS) in microtitration...
Lomholt, Jeanet Andersen; Kilian, Mogens
from patients with keratitis caused by P. aeruginosa, Streptococcus group G, Moraxella catarrhalis, Staphylococcus aureus, coagulase-negative staphylococci, and the IgA1 protease-producing Streptococcus pneumoniae were compared with S-IgA in tear fluid, colostrum, and saliva from healthy individuals...
Nielsen, Stig Lønberg; Pedersen, C; Jensen, T G
) and the incidence rate of nosocomial bacteremia decreased by 28.9% from 82.2 to 56.0 (4.2% annually, p Staphylococcus aureus (12.3%), coagulase-negative staphylococci (10.......0%) and Streptococcus pneumoniae (9.1%). Regardless of place of acquisition, the proportion of bacteremias caused by enterococci increased (p
The screening of multi-drug resistance (MDR) susceptibilities of Staphylococcus aureus and Staphylococcus epidermidis to methicillin and vancomycin in teaching ... β-lactam antibiotics that predispose patients to acquisition of methicillin -resistant Staph. aureus (MRSA) and coagulase negative resistant staphylococci.
van der Zwet, Wil C.; Debets-Ossenkopp, Yvette J.; Reinders, Erik; Kapi, Maria; Savelkoul, Paul H. M.; van Elburg, Ruurd M.; Hiramatsu, Keiichi; Vandenbroucke-Grauls, Christina M. J. E.
A premature infant in a neonatal intensive care unit (NICU) developed a bloodstream infection caused by coagulase-negative staphylococci (CoNS) sensitive to vancomycin. The infection persisted for 3 weeks, despite therapy with vancomycin and replacement of all intravenous catheters. The neonate died
region or upon the device2. The agent is usually one of the various microorganisms settled on the skin flora of which Staphylococcus aureus and coagulase negative staphylococci are the most probable candidates3. In this case an ICD pocket infection caused by Klebsiella pneumonia, which is a rare cause, is presented.
Oct 22, 2014 ... results show that lactic acid bacteria (LAB) and coagulase negative staphylococci (CNS) represent the two predominant microflora in all ... and it contains all of the essential amino acids for human consumption (Varnam and ... fermented sausages are produced with starter cultures combination of lactic acid ...
Nielsen, Xiaohui Chen; Chen, Ming; Hellesøe, Anne-Marie Blok
We conducted a retrospective epidemiologic study of catheter related bloodstream infections (CRBSI) in patients receiving long-term home parenteral nutrition (HPN) from January 2002 to December 2005. Our results showed that coagulase negative staphylococci (CoNS) were the most prevalent pathogens...
Oct 22, 2014 ... The aim of this work was to study the effect of Staphylococcus xylosus and Lactobacillus plantarum on microbial and physicochemical characteristics of a dry fermented poultry meat sausage. The microbial results show that lactic acid bacteria (LAB) and coagulase negative staphylococci (CNS) represent ...
McDivitt, Maxine E.; Topp, Eleanor B.
Six coagulase-positive strains of Staphylococcus aureus which had been cultivated in Brain Heart Infusion broth, milk, and brine were plated on seven isolation media. A significant difference in the growth patterns of the individual strains was found as well as a significant effect resulting from the previous cultivation history before plating. Brine and, to a lesser extent, milk were found to reduce maximal cell concentrations attained, but strains grown in brine and milk showed greater ability to withstand the selective action of the isolation media. Fibrinogen applied to the surface of five of the media allowed the formation of characteristic halos by coagulase-positive strains of S. aureus. Only half of the strains studied produced a zone of precipitation on SM110-Egg Yolk agar. The isolation medium containing cycloheximide and a high level of polymxin B was most inhibitory to the organisms. PMID:14131367
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
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 Â© 2012 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.
Penna, Bruno; Varges, Renato; Medeiros, Luciana; Martins, Gabriel M; Martins, Rodrigo R; Lilenbaum, Walter
The diversity of species of the genus Staphylococcus sp. and the antimicrobial resistance of isolates from 151 unmedicated dogs of both sexes with a clinical diagnosis of otitis were recorded. Ninety-one isolates of Staphylococcus spp. were identified by biochemical reactions and tested for susceptibility to 15 antimicrobials. Coagulase-positive species were most common; S. pseudintermedius (38.4%), S. schleiferi schleiferi (15.4%), S. aureus (14.3%), S. epidermidis (11%), S. simulans (11%), S. schleiferi coagulans (8.8%) and S. saprophyticus (1.1%). All the isolates showed resistance to at least one drug and 89% were multiresistant. Amoxicillin combined with clavulanic acid and oxacillin were the most effective, while resistance was widely observed for neomycin and erythromycin. The results highlight the recognition and the potential need for bacterial culture with species identification and antimicrobial susceptibility tests for appropriate antimicrobial therapy.
Sukhanova, S M; Zakharova, N E; Golubenko, I A
Comparative trials were carried out new commercial nutrient media made by Russian (Mirogen Research-and-Production Association, Ministry of Health of the Russian Federation; State Research Center for Applied Microbiology and Biotechnology, Obolensk, Serpukhov District, Moscow Region; and I. I. Mechnikov Biomed, Moscow Region; Research Center of Pharmacotherapy, Saint Petersburg) and foreign (NovaMed Ltd, Israel) manufacturers to isolate staphylococci. Their quality was assessed in terms of physicochemical and biological parameters. The test nutrient media provided a good growth of specifically important test staphylococcal strains and inhibited Enterobacteriaceae. The technique of using the traditional inoculum to Petri dish ensures a higher sensitivity and inhibition of nutrient media as compared to that of applying the inoculum to NovaStreak.
Michelle M. S. Chen
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.
Namvar, Amirmorteza Ebrahimzadeh; Bastarahang, Sara; Abbasi, Niloufar; Ghehi, Ghazaleh Sheikhi; Farhadbakhtiarian, Sara; Arezi, Parastoo; Hosseini, Mahsa; Baravati, Sholeh Zaeemi; Jokar, Zahra; Chermahin, Sara Ganji
Staphylococci are known as clustering Gram-positive cocci, nonmotile, non-spore forming facultatively anaerobic that classified in two main groups, coagulase-positive and coagulase-negative. Staphylococcus epidermidis with the highest percentage has the prominent role among coagulase-negative Staphylococci that is the most important reason of clinical infections. Due to various virulence factors and unique features, this microorganism is respected as a common cause of nosocomial infections. Because of potential ability in biofilm formation and colonization in different surfaces, also using of medical implant devices in immunocompromised and hospitalized patients the related infections have been increased. In recent decades the clinical importance and the emergence of methicillin-resistant Staphylococcus epidermidis strains have created many challenges in the treatment process. PMID:25285267
Riber, U; Espersen, F; Skinhøj, P
The ability of staphylococci adherent to silicone surfaces to induce superoxide anion (O2-) production by polymorphonuclear leukocytes (PMNs) was investigated and compared with the same activity induced by planktonic bacteria. The responses to Staphylococcus aureus strain E 2371 and Staphylococcus...... epidermidis strain ATCC 14990 were compared. The staphylococci were allowed to adhere to silicone catheters for 2 h at 37 degrees C. After opsonization of adherent bacteria in 30% human AB-positive serum, the induction of superoxide anion production by PMNs was measured in a cytochrome C reduction assay. Both...... bacterial strains, when adhered to the surfaces, were able to induce superoxide anion production by PMNs to about the same extent. Comparing adherent and planktonic bacteria with these two bacterial strains, it was found that planktonic S. epidermidis induced one to three times higher superoxide anion...
Watts, J L; Pankey, J W; Nickerson, S C
The Staph-Ident and STAPHase systems (Analytab Products, Plainview, N.Y.) were compared with conventional methods for identification of staphylococci isolated from bovine intramammary infections. Adjunct testing by colony morphology, pigmentation, and biochemical tests was conducted to resolve discrepant identifications. The initial accuracies of the conventional scheme and Staph-Ident were 92.1 and 89.2%, respectively. Staphylococcus hyicus subsp. chromogenes could not be identified by means...
Oliveira Lis Christina de; Leite Carlos Artur Lopes; Brilhante Raimunda Sâmia N.; Carvalho Cibele Barreto M.
This study evaluated the middle ear microbiota and antimicrobial susceptibility patterns from strains isolated from dogs with otitis media. A total of 62 dogs obtained from Zoonoses Control Center of Fortaleza City Ceará State / Brazil were studied over a 10-month period (August/2003 to June/2004). Of the total, 46.8% (n=30) of the animals were positive for otitis media and the infection was monomicrobial in 76.6% of them. The most frequent isolated agents were coagulase-positive Staphylococc...
Joyce Bruna Paiva Silva
Full Text Available Goat’s milk is an excellent food that can guarantee beneficial effects to human health, but its characteristics are easily altered by the action of microorganisms and by manipulation. The present study aimed to evaluate the microbiological quality of goat milk in 11 farms in the region of Macaíba/RN. Samples of milk and swabs from the hands of milkers, utensils and goat teats were collected and transported in a thermal box (10 ºC – 12 ºC to the Laboratory of Food Engineering of UFRN. The microorganisms studied were the aerobic mesophilic bacteria, psychrotrophic, coagulase positive and negative staphylococci, coliforms at 35 ºC and 45 ºC. Regarding the hands of milkers, 56% presented contamination by aerobic mesophilic bacteria. In the analyzed utensils were found, in 18.2%, coliforms at 45 ºC and coagulase positive staphylococci, as well as coagulase negative staphylococci (63.6% and coliforms at 35 ºC in 27.3% of the samples. For cold raw goat milk, 10% of the samples presented contamination with psychrotrophic and 17% with coliforms at 45 ºC. Regarding goat teats, 52% of the samples had aerobic mesophilic bacteria counts above 103 CFU/ceiling, 61% presented coagulase negative staphylococci and 10% coagulase positive staphylococci, this one of the main agents causing mastitis. Thus, these values indicate that hygienic procedures need to be adopted appropriately in the milking of raw goat milk, requiring guidelines that enable producers to obtain higher quality milk.
Full Text Available The objective of this study was to determine the distribution of genes encoding aminoglycoside-modifying enzymes (AMEs and staphylococcal cassette chromosome mec (SCCmec elements among clinical isolates of methicillin-resistant staphylococci (MRS. Antibiotic susceptibility test was done using Kirby-Bauer disk diffusion method. The presence of SCCmec types and AME genes, namely, aac (6′-Ie-aph (2′′, aph (3′-IIIa and ant (4′-Ia was determined using two different multiplex polymerase chain reaction. The most encountered AME genes were aac (6′-Ie-aph (2′′ (55.4% followed by aph (3′-IIIa (32.3% and ant (4′-Ia gene (9%. SCCmec type I (34% was predominant in this study. In conclusion, the aac (6′-Ie-aph (2′′ was the most common AME gene and SCCmec type I was most predominant among the MRS isolates.
Sherman, Erica; Moormeier, Derek; Bayles, Kenneth; Wei, Timothy
Staphylococci bacteria are recognized as the most frequent cause of biofilm-associated infections. A localized staph infection has the potential to enter the bloodstream and lead to serious infections such as endocarditis, pneumonia, or toxic shock syndrome. Changes in flow conditions, such as shear stress, can lead to stable biofilm growth or the dispersion of portions of the biofilm downstream. Exploration of biofilm physiology indicates a link between production of a specific enzyme called nuclease and biofilm architecture -; however the physical impact of this enzyme in directing the location and behavior of biofilm growth remains unclear. This talk investigates the link between sites of nuclease production and the development of biofilm tower structures using the application of micro-PIV and fluorescently labeled bacterial cells producing nuclease. Staphylococcus aureus bacteria were cultured in a BioFlux1000 square microchannel of a 65 by 65 um cross section, and subjected to a steady shear rate of 0.6 dynes. Micro-PIV and nuclease production measurements were taken to quantify the flow over a biofilm tower structure prior and during development. Data were recorded around the structure at a series of two dimensional planes, which when stacked vertically show a two dimensional flow field as a function of tower height.
Sienkiewicz, Monika; Głowacka, Anna; Poznańska-Kurowska, Katarzyna; Kaszuba, Andrzej; Urbaniak, Anna; Kowalczyk, Edward
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.
Full Text Available Objective(s: Staphylococcus aureus is an important bacterial pathogen responsible for a variety numbers of nosocomial and community acquired infections. Biofilm formation is regarded as an important factor in the establishment of S. aureus infection. The contribution of the genetic background of S. aureus to biofilm formation is poorly understood. The aim of the present work was to genotype S. aureus strains associated to biofilm based on the coagulase and protein A genes and to evaluate the association between the genetic background and the biofilm forming ability of clinical S. aureus isolates. Materials and Methods: A total number of 100 S. aureus were isolated from nosocomial infections and biofilm formation capability was investigated using phenotypic assay and molecular detection of biofilm associated genes. The strains were genotyped based on coagulase (coa and protein A (spa gene polymorphisms using restriction fragments length polymorphism-polymerase chain reaction (RFLP-PCR. Results: RFLP-PCR of coa gene generated two types and three subtypes. Amplification of spa gene resulted in two banding patterns and their restriction digestion generated three subtypes. The combined coa and spa RFLP patterns generated nine genotypes (G1-G9. The genotypes G4 and G1 were the most prevalent (32.1% and 24.3%, respectively. Conclusion: High clonal diversity of S. aureus strains able to produce biofilm was observed. Biofilm formation correlates with the spa and coa clonal lineage in our population and testing for multiple gene polymorphisms could be employed for local epidemiologic purposes.
Zapotoczna, Marta; McCarthy, Hannah; Rudkin, Justine K; O'Gara, James P; O'Neill, Eoghan
High-level resistance to antimicrobial drugs is a major factor in the pathogenesis of chronic Staphylococcus aureus biofilm-associated, medical device-related infections. Antimicrobial susceptibility analysis revealed that biofilms grown for ≤ 24 hours on biomaterials conditioned with human plasma under venous shear in iron-free cell culture medium were significantly more susceptible to antistaphylococcal antibiotics. Biofilms formed under these physiologically relevant conditions were regulated by SaeRS and dependent on coagulase-catalyzed conversion of fibrinogen into fibrin. In contrast, SarA-regulated biofilms formed on uncoated polystyrene in nutrient-rich bacteriological medium were mediated by the previously characterized biofilm factors poly-N-acetyl glucosamine, fibronectin-binding proteins, or autolytic activity and were antibiotic resistant. Coagulase-mediated biofilms exhibited increased antimicrobial resistance over time (>48 hours) but were always susceptible to dispersal by the fibrinolytic enzymes plasmin or nattokinase. Biofilms recovered from infected central venous catheters in a rat model of device-related infection were dispersed by nattokinase, supporting the important role of the biofilm phenotype and identifying a potentially new therapeutic approach with antimicrobials and fibrinolytic drugs, particularly during the early stages of device-related infection. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: email@example.com.
A multiplex PCR method was developed for simultaneous detection of the genes encoding methicillin resistance (mecA), staphylococcal enterotoxins A, B and C (sea, seb and sec), coagulase (coa) and 16S rRNA. The primers for amplification of the 16S rRNA gene were specific for Staphylococcus spp., and ...
Sturm, P.D.J.; Kwa, D.; Vos, F.J.; Bartels, C.J.; Schulin, T.
Test parameters and clinical impact of the direct tube coagulase test (DTCT) for rapid identification of Staphylococcus aureus from blood culture were investigated. The sensitivity of the DTCT at 4 h using saline dilution was 96%, compared with 93% using serum separator tubes; specificity was 100%
R. P. Ratti
Full Text Available Endophytic microorganisms are relatively unstudied as potential sources of novel natural products for medical and commercial exploitation. The aim of this work was to investigate some Brazilian tropical savannah trees Cassia leptophylla and Prunus spp. in order to isolate the endophytic microorganisms associated with these plants. The samples were disinfected to eliminate the epiphytic population. Colonies were diluted and displayed as drops in media and growing colonies were inactivated. Staphylococcus coagulase-positive strain was used as indicator microorganism and subjected to the antibioses test. Data showed that the microorganisms isolated from Cassia leptophylla had no inhibition against Staphylococcus. On the other hand, microorganisms isolated from Prunus spp. leaves showed antibacterial activity and inhibited Staphylococcus when cultivated in peptone agar as well as in yeast extract agar. Investigation proceeds in order to classify the isolated microorganisms presenting bioactive substance and exploit the potential of the compounds produced to inhibit the indicator bacteria. Other bioactive properties will be investigated.
Hamiroune, M; Berber, A; Boubekeur, S
The authors describe a survey and screening programme for staphylococcus. The study covers 14 dairy farms in the Algiers region, from which 203 samples of cows' milk were taken for bacteriological testing. The survey results show that poor husbandry conditions are the main cause of staphylococcus in cows' milk. Staphylococcus was found in the milk of 30% of the cows sampled. These results were influenced by a variety of factors, in that: the contamination rate rose with the number of pregnancies, age, and volume of milk output of the cow, as well as the bedding thickness; the milk contamination rate was greater when milking occurred outside a milking parlour and when it was performed by machine; higher rates of staphylococcus infection were found in the milk of cows at the end of lactation, in red and white breeds, and in those with cylindricalteats. Identification of the bacteria found (staphylococcus) showed that coagulase- negative staphylococci were present in 67.21% of samples, whereas coagulase- positive staphylococci were present in only 32.79%. The average count for the latter was equal to 0.54 x 10(4) colony-forming units per ml of Staphylococcus aureus. Seventy percent of the milk analysed was free from staphylococci and most of the bacteria identified were not pathogenic to consumers (coagulase- negative staphylococci); nevertheless, consuming fresh milk still presents a degree of risk.
Andre Ricardo Araujo da Silva
Full Text Available OBJETIVO: Avaliar as infecções relacionadas à assistência à saúde, em unidade de terapia intensiva neonatal, causadas pelo Staphylococcus coagulase negativa, verificando o perfil de sensibilidade antimicrobiana e possíveis esquemas antibióticos eficazes. MÉTODOS: Estudo descritivo retrospectivo de uma série de casos de infecções relacionadas à assistência à saúde tardias de origem hospitalar atribuída ao Staphylococcus coagulase negativa, avaliando o perfil de sensibilidade antimicrobiana. Foram estudados os recém-nascidos internados entre 1º de janeiro de 2010 a 30 de junho de 2012 em uma unidade de terapia intensiva neonatal da cidade do Rio de Janeiro, sendo todos os pacientes oriundos de outras unidades. RESULTADOS: Foram admitidos 765 pacientes, totalizando 3.051 pacientes-dia e uma densidade de incidência de infecção geral de 18,9 por 1.000 pacientes-dia. A taxa de utilização de cateteres venosos centrais foi de 71,6% e a positividade das culturas de todos os sítios para todas as infecções relacionadas à assistência à saúde foi de 68,4%. O Staphylococcus coagulase negativa foi implicado em 11 (19,2% das 57 infecções relacionadas à assistência à saúde e Klebsiela pneumoniae produtor de betalactamase de espectro estendido e Candida sp em 5 ocasiões cada. Das 11 infecções, 10 (90,9% foram atribuídas a infecções primárias de corrente sanguínea. A sensibilidade dos isolados de Staphylococcus coagulase negativa em relação à vancomicina, clindamicina, ciprofloxacin, oxacilina e gentamicina foi de 100%, 81,8%, 72,7%, 27,2%, 22,2%, respectivamente. Não houve óbito atribuído diretamente à infecção por Staphylococcus coagulase negativa. CONCLUSÃO: O Staphylococcus coagulase negativa foi o principal agente encontrado nas infecções relacionadas à assistência à saúde tardias de origem hospitalar, sendo baixas as taxas de infecções relacionadas a cateter venoso central. Em hospitais
E. L. Pizzolitto
Full Text Available
A produção de slime é um importante fator de virulência dos estafilococos coagulase-negativa, permitindo-lhes aderir sobre as superfícies lisas de biomateriais, e por isso, é associada aos processos de infecção de implantes. No presente estudo a produção de slime em 27 cepas de estafilococos coagulase-negativa foi investigada por cultura em ágar vermelho Congo (77,7% de positividade, método espectrofotométrico ou da microplaca (81,4% de positividade e microscopia eletrônica de varredura (88,9% de positividade. Foi também avaliada a resistência de estafilococos coagulasenegativa a vários antimicrobianos usando a técnica do disco difusão. A porcentagem de resistência à penicilina G, oxacilina, eritromicina, clindamicina e gentamicina em estafilococos produtores de slime foi respectivamente de 88,9%; 70,4%; 81,5%; 66,7% e 59,2%; todos os estafilococos coagulase-negativa foram vancomicina sensíveis. As cepas isoladas de cateter venoso central foram identificadas por método convencional e sistema API Staph. Os 27 estafilococos coagulase-negativa foram identificados como: S. saprophyticus (3,7%, S. xylosus (7,4%, S. haemolyticus (14,8%, S. epidermidis (37,0%, S. warneri (14,8%, S. lugdunensis (7,4%, S. hominis (7,4%, S. schleiferi (3,7% e S. chromogenes (3,7%. Pode-se concluir que entre a maioria das espécies Staphylococcus coagulase-negativa houve associação entre a produção de slime, origem nosocomial das cepas e reduzida sensibilidade aos antimicrobianos, sugerindo potencial patogênico no ambiente hospitalar. Palavras-chave: estafilococos coagulase-negativa; slime; fatores de virulência; polissacarídeo extracelular; biofilme; sensibilidade aos antimicrobianos.
Desenvolvimento de material de referência para microbiologia de alimentos contendo estafilococos coagulase positiva em matriz queijo Development of reference material for the microbiology of foods containing coagulase-positive Staphylococcus in a cheese matrix
Marcelo Luiz Lima Brandão
Full Text Available O uso de materiais de referência (MR é uma das principais ferramentas utilizadas para garantia e controle da qualidade de laboratórios de microbiologia de alimentos. No Brasil, a RDC n.º 12/01 da Anvisa prevê como um dos parâmetros para a avaliação da qualidade de queijos a enumeração de estafilococos coagulase positiva (ECP. O grande desafio na produção de MR destinados a ensaios microbiológicos é a instabilidade natural dos micro-organismos, o que dificulta o desenvolvimento e a manutenção desses MR. O objetivo deste estudo foi produzir um MR quantitativo destinado ao ensaio de enumeração de ECP em matriz queijo. Uma amostra de queijo ultrafiltrado com contagem de ECP The use of reference materials (RM is one of the principal tools used for assurance and quality control in food microbiology laboratories. In Brazil, Anvisa RDC nº 12/01 specifies the enumeration of coagulase-positive staphylococcus (CPS as one of the parameters for evaluating cheese quality. The main challenge in the production of RM destined for microbiological assays is the natural instability of the microorganisms, which makes it difficult to develop and maintain them. This study aimed to produce a quantitative RM for use in enumeration assays of CPS in cheese matrixes. A sample of an ultra-filtered cheese with a CPS count of <10 CFU/g and a total nº of viable aerobes of 1.2 × 10³ CFU/g was used as the matrix to produce the RM. The matrix was distributed in flasks, contaminated with specific concentrations of the target bacteria and submitted to freeze-drying. Sucrose was used as the cryo-protector. The RM produced was considered homogeneous and stable at < -70ºC during the entire study period (10 months. The material was shown to be stable at 4, 25, 30 and 35 ºC for 4 days, although the results indicated a decrease in cell concentration at 35 ºC. At -20 ºC the RM was stable for 48 days. It was concluded that the material showed all the necessary
Anita Dwi Anggraini
Full Text Available Abstract: Analysis of the polymorphism of a gene is important to obtain early information in identifying genetic markers related to the characteristics to be seen. The RFLP method becomes one of the chosen methods because it can see polymorphism that can be detected by using the different fragments of DNA that have been cut by using certain endonuclease enzyme so that it is possible to describe the polymorphism of a gene. Application of RFLP in this study was devoted to see the coagulase gene polymorphism of methichillin-resistant S. aureus. Based on the description above, researcher was interested in conducting the analysis of CoA (coagulase gene polymorphism with AluI restriction site of methicillin-resistant S. aureus isolates. The results of this study are aimed to be scientific information about the genetic variation of the coagulase gene of methicillin-resistant S. aureus, and as information in the management of diseases related to antibiotic resistance. This was a descriptive study intended to discover the coagulase gene polymorphism of methichillin-resistant S. aureus. The research was conducted in Microbiology Laboratory of Health Analyst Department, Surabaya, and Human Genetic Laboratory of Institute of Tropical Disease of Airlangga University Surabaya. Samples from this study was a collection of isolates of MRSA in Microbiology Laboratory of Dr. Soetomo Hospital Surabaya selected by using consecutive sampling. Based on the bacteriological examination, all of the samples were S. aureus and the results of amplification showed that PCR product (amplicon of mecA and CoA gene from specific primers, that were 304 bp and 756 bp, had a positivity of 100%. The results of PCR-RFLP of CoA gene showed that all 25 samples underwent polymorphism which was divided into four RFLP patterns and the biggest one was RFLP I pattern (the one which was not digested by AluI restriction enzyme with a proportion of 64%.
Comparative evaluation of two matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems, Vitek MS and Microflex LT, for the identification of Gram-positive cocci routinely isolated in clinical microbiology laboratories.
Lee, Miae; Chung, Hae-Sun; Moon, Hee-Won; Lee, Sun Hwa; Lee, Kyungwon
We evaluated the performance of two MALDI-TOF MS systems for the identification of clinically important Gram-positive cocci. Vitek MS and Microflex LT correctly identified 97.2% and 94.7%, respectively. Both systems offer reliable and rapid identification of clinically important Gram-positive cocci isolated in clinical laboratories, including staphylococci, streptococci, and enterococci. Expanding the databases, especially of coagulase-negative staphylococci and viridans streptococci, would enhance performance. Copyright © 2015 Elsevier B.V. All rights reserved.
Morris, Daniel O; Boston, Raymond C; O'Shea, Kathleen; Rankin, Shelley C
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.
Swart, Henriëtte de; Sag, I.A.
This paper addresses the two interpretations a combination of negative indefinites can get in concord languages like French, namely a concord reading which amounts to a single negation, or a double negation reading. We develop an analysis in a polyadic framework, in which a sequence of
Møller, Maria E.; Alanin, Mikkel C.; Grønhøj, Christian
, and microbiology analyses from the nose or paranasal sinuses. Results: We included 46 studies (1823 patients) from 16 countries. Staphylococcus aureus was found in 30% of the noses and sinuses of patients with CF. Other common bacteria found included Pseudomonas aeruginosa, coagulase negative staphylococci......, and Haemophilus influenzae. In PCD, H. influenzae was the most common bacteria (28%), followed by Streptococcus pneumoniae and P. aeruginosa. If studies that included nonsurgical swab and blowing samples were excluded, then P. aeruginosa was the most common bacterium in patients with CF (34%) and in patients...... with PCD (50%), followed by S. aureus and H. influenza. Conclusion: S. aureus, P. aeruginosa, coagulase negative staphylococci, and H. influenzae dominated in the upper airways of patients with CF. In patients with PCD, H. influenzae, S. pneumoniae, and P. aeruginosa dominated. When studies that included...
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
Zhao, Yanqing; Liu, Hui; Zhao, Xuanduo; Gao, Yang; Zhang, Miaotao; Chen, Dekun
Subclinical mastitis, a costly disease for the dairy industry, is usually caused by intramammary bacterial infection. The aim of this study was to investigate the prevalence of and pathogens involved in subclinical mastitis in dairy goats in China. A total of 683 dairy goats in the main breeding areas of China were selected, and milk samples were collected. Out of these, 313 (45.82 %) goats were detected distinct or strong positive for subclinical mastitis by using California mastitis test. Among these positive goats, 209 milk samples were used to identify the causing agents by a multiplex PCR assay, and results were listed as follows: coagulase-negative staphylococci (59.52 %), Staphylococcus aureus (15.24 %), Escherichia coli (11.43 %), and Streptococcus spp. (10.95 %). In conclusion, subclinical mastitis is a highly prevalent disease in dairy goats in China, and coagulase-negative staphylococci are the predominant pathogens.
Full Text Available Coagulase-positive Staphylococcus aureus is introduced as the third major source of animal origin food borne disease. This bacterium can proliferate and produce toxin in protein and carbohydrate foods leading to food-poisoning. The aim of this study was isolation, identification and enumerating of S. aureus in meat as well as intestinal contents of native duck around Tabriz area. For this purpose, a total number of 35 native ducks randomly was purchased in the nearby villages of Tabriz. Fifty g of thigh meat together with 10 g of intestinal content of each carcass were sampled and analyzed according to the ISIRI protocols. The isolates were confirmed by PCR technique. Results showed that 17.14% of meat and fecal samples were found contaminated with S. aureus. Mean values of S. aureus load in meat and feces were estimated at 87 CFU/g and 64 CFU/g, respectively. It was assumed that contamination of duck meat with S. aureus can be occurred through non-hygienic practices during slaughter as well as contamination with intestinal contents and/or skin of the carcass.
Koçoğlu, Esra; Karabay, Oğuz
The coagulase-negative bacterial species Staphylococcus sciuri is widely distributed in the natural environment. Although principally found in animals, S. sciuri is occasionally isolated from human samples. In this paper, S. sciuri bacteremia which was associated with an indwelling catheter of a patient with acute myeloid leukemia (AML) and neutropenia was presented. An empirical intravenous antibiotic therapy (meropenem, vancomycin) was initiated with the preliminary diagnosis of febrile neutropenia and catheter infection. The catalase and oxidase positive, tube coagulase negative strain isolated from three of the concurrent blood cultures and intravenous catheter culture has been identified as S. sciuri. The isolate was found resistant to penicilin and oxacilline. This case has emphasized the importance of identification of coagulase-negative staphylococci isolated from the cultures of patients with haematological malignancy.
A new agglutination test (Monostaph +; Bionor, Skien, Norway) has been developed. This new agglutination test has been compared with two other agglutination tests for the identification of 128 isolates of Staphylococcus aureus and 82 coagulase-negative staphylococci. The sensitivities of both Monostaph + and Pastorex Staph-Plus were excellent (98.7 and 97.4%, respectively) in detection of oxacillin-resistant Staphylococcus aureus. The specificity was 96.4% (two Staphylococcus epidermidis isol...
Gregson, D B; Low, D E; Skulnick, M; Simor, A E
Six rapid agglutination tests for identification of Staphylococcus aureus were evaluated by using 62 strains of S. aureus, 63 strains of S. saprophyticus, and 67 strains of other coagulase-negative staphylococci. S. saprophyticus was responsible for 19 of 26 false-positive results and 20 uninterpretable reactions. Thus, urinary staphylococcal isolates that are positive by rapid agglutination tests may require other confirmatory tests for the identification of possible S. saprophyticus.
Doyle, P W; Woodham, J D
In a prospective study, patients with the diagnosis of chronic ethmoid sinusitis were evaluated microbiologically by using biopsy specimens of the ethmoid sinus mucosa. Microbiology cultures were performed on 94 specimens from 59 patients. Staphylococcus aureus and members of the family Enterobacteriaceae were the most frequent classical pathogenic bacteria isolated. Coagulase-negative staphylococci were the most common overall isolates. Streptococcus pneumoniae and Haemophilus influenzae wer...
Jana Pukáčová; Lucia Poľaková; Eva Dudriková
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 %) wer...
Marull, Jorge; Casares, Pablo A
Staphylococcus aureus, Coagulase-negative staphylococci, and Enterococci sp. are the usual pathogens involved in nosocomial bacterial endocarditis. Corynebacterium species isolation in blood specimens is usually considered to be a contaminant. We present an interesting case of native mitral valve endocarditis in a 73 year old African American female that was diagnosed days after she was discharged from our institution. The infection was cleared with medical therapy alone.
Marull, Jorge; Casares, Pablo A
Staphylococcus aureus, Coagulase-negative staphylococci, and Enterococci sp. are the usual pathogens involved in nosocomial bacterial endocarditis. Corynebacterium species isolation in blood specimens is usually considered to be a contaminant. We present an interesting case of native mitral valve endocarditis in a 73 year old African American female that was diagnosed days after she was discharged from our institution. The infection was cleared with medical therapy alone.
Fomsgaard, A; Høiby, Niels; Friis, H M
in the other countries. Staphylococcus aureus was the most frequent wound isolate in Denmark, while Enterobacteriaceae other than Escherichia coli dominated in other European countries. Thus, in Denmark Escherichia coli and Staphylococcus aureus, followed by Pseudomonas aeruginosa and Klebsiella spp. (from...... ICUs) or Enterococcus spp. and Klebsiella spp. (from hematology/oncology units), are the most prominent pathogens in these units today. Indicator organisms of antibiotic consumption (Pseudomonas aeruginosa and methicillin-resistant coagulase-negative staphylococci and Staphylococcus aureus) were more...
The distribution of various pathogens causing wound infection was evaluated in Bharatpur, Chitwan, Nepal. A total of 200 pus specimens were cultured, of which 75% showed bacterial growth. Ten different species of bacteria were isolated. Staphylococcus aureus was the most common organism [65.3%] followed by Klebsiella pneumoniae [8%], Escherichia coli [7.3%], Coagulase negative staphylococci [6%], Pseudomonas aeruginosa [5.3%], Enterococcus species [3.3%], Enterobacter species [2%], Acinetob...
Horslund, Camilla Søballe
to semantics and pragmatics, negations can be used in three different ways, which gives rise to a typology of three different types of negations: 1) the descriptive negation, 2) the polemic negation, and 3) the meta-linguistic negation (Nølke 1999, 4). This typology illuminates the fact that the negation...... as such may be more or less central to the meaning of the utterance. The present paper investigates the role of morphosyntactic and prosodic prominence as well as register and social setting on the interpretation of negations. It seems plausible to expect that if the negation as such is central to the meaning...... of the utterance (as in polemic negations), the negation will be articulated prominently in order to emphasise this importance. Likewise, if the negation is not central to the meaning of the utterance, it should not be articulated prominently. Moreover, it is plausible to expect descriptive negations to be more...
Fahmi Yousef Khan
Full Text Available Background and Objectives. Bacterial meningitis is a common medical condition in Qatar. The aim of this study was to describe the clinical characteristics of bacterial meningitis, the frequency of each pathogen, and its sensitivity to antibiotics and risk factors for death. Patients and Methods. This retrospective study was conducted at Hamad General Hospital between January 1, 2009, and December 31, 2013. Results. We identified 117 episodes of acute bacterial meningitis in 110 patients. Their mean age was 26.4±22.3 years (range: 2–74 and 81 (69.2% of them were male patients. Fifty-nine episodes (50.4% were community-acquired infection and fever was the most frequent symptom (94%, whereas neurosurgery is the most common underlying condition. Coagulase-negative staphylococci were the most common causative agent, of which 95% were oxacillin-resistant, while 63.3% of Acinetobacter spp. showed resistance to meropenem. The in-hospital mortality was 14 (12%. Only the presence of underlying diseases, hypotension, and inappropriate treatment were found to be independent predictors of mortality. Conclusion. Acute bacterial meningitis predominantly affected adults and coagulase-negative staphylococci species were the common causative agent in Qatar with majority of infections occurring nosocomially. More than 90% of all implicated coagulase-negative staphylococci strains were oxacillin-resistant.
Soares, Andréa Ferreira; Aquino, Ana Rafaela Luz de; Carvalho, Cyntia Helena Pereira de; Nonaka, Cassiano Francisco Weege; Almeida, Dulce; Pinto, Leão Pereira
In view of the morbidity potential of oral complications in patients with leukemia, this study evaluated the clinical and microbiological alterations that occur in the oral mucosa of children with acute lymphoblastic leukemia (ALL) undergoing antineoplastic chemotherapy and prophylactic administration of 0.12% chlorhexidine gluconate. The sample consisted of 17 children aged 2 to 12 years that underwent clinical examination of the oral mucosa for the detection of oral lesions. In addition, biological material was collected from labial and buccal mucosa for microbiological analysis. Oral mucositis was observed in only 5 (29.4%) patients. Microbiological analysis revealed a reduced number of potentially pathogenic microorganisms, such as coagulase-negative staphylococci (47%), Candida albicans (35.3%), Klebsiella pneumoniae (5.9%), enteropathogenic Escherichia coli (5.9%), and Stenotrophomonas maltophilia (5.9%). Patients with oral mucositis showed a higher frequency of coagulase-negative staphylococci (80%) when compared with patients with normal oral mucosa (33.3%). In conclusion, the results of the present study suggest that the prophylactic use of 0.12% chlorhexidine gluconate reduces the frequency of oral mucositis and oral pathogens in children with ALL. In addition, the present findings suggest a possible relationship between coagulase-negative staphylococci and the development of oral mucositis.
Rosanna ePapa; Laura eSelan; Ermenegilda eParrilli; Marco eTilotta; Filomena eSannino; Georges eFeller; Maria Luisa eTutino; Marco eArtini
Microbial biofilms have great negative impacts on the world’s economy and pose serious problems to industry, public health and medicine. The interest in the development of new approaches for the prevention and treatment of bacterial adhesion and biofilm formation has increased. Since, bacterial pathogens living in biofilm induce persistent chronic infections due to the resistance to antibiotics and host immune system. A viable approach should target adhesive properties without affecting bacte...
A. L. L. CORTEZ
Full Text Available
Lingüiça frescal faz parte dos alimentos expostos à contaminações e constitue um excelente meio para desenvolvimento e multiplicação de microrganismos. Assim, a presente pesquisa teve como objetivo verificar a qualidade microbiológica de 106 amostras de lingüiça frescal de carne de frango, mista (carne bovina e suína e de carne suína, fabricadas artesanalmente e sob inspeção, no município de Jaboticabal – SP, Brasil. Determinou-se o número mais provável de coliformes fecais por grama (NMP/g do produto e a ocorrência de Escherichia coli, contagem de estafilococos coagulase-positiva (ECP, pesquisa de Salmonella spp. e Campylobacter spp.. Foram utilizados métodos microbiológicos convencionais a fim de detectar, isolar e identificar estas bactérias, comparando os resultados obtidos com os limites estabelecidos pela legislação brasileira. Das 106 amostras analisadas, 78 (73,6% demonstraram a presença de coliformes fecais, 41 (38,7% de Escherichia coli, 18 (17% de ECP e 8 (7,5% de Salmonella spp.. Nenhuma amostra foi positiva para Campylobacter spp. Os testes da PCR multiplex rfb, stx1, stx2, eae O157, O113, O111 das amostras de E. coli foram negativos. Em decorrência dos resultados obtidos pode-se concluir que se faz necessária melhoria nas condições higiênico-sanitárias desse tipo de produto.
Enumeration of coagulase and thermonuclease-positive Staphylococcus spp. in raw milk and fresh soft cheese: an evaluation of Baird-Parker agar, Rabbit Plasma Fibrinogen agar and the Petrifilm Staph Express count system.
Viçosa, Gabriela Nogueira; Moraes, Paula Mendonça; Yamazi, Anderson Keizo; Nero, Luís Augusto
Staphylococcus spp. are microorganisms that are naturally present in milk and dairy products and are often associated with food-borne diseases outbreaks due to the ability of some strains to produce thermostable enterotoxins. This ability is usually associated with coagulase and thermonuclease production, characteristics that are considered in the microbiological analyses for the control of such microorganisms. The objective of this study was to evaluate the culture media and the methodologies used for the enumeration of coagulase and thermonuclease-positive Staphylococcus spp. in raw milk and fresh soft cheese. Samples of artificially contaminated milk (with coagulase-positive Staphylococcus reference strains) and samples of naturally contaminated raw milk and cheese were submitted for enumeration in Baird-Parker agar (BP), Rabbit Plasma Fibrinogen agar (RPFA) and in the Petrifilm Staph Express count system (STX). No significant differences (P > 0.05) were observed between the mean counts obtained in all of the evaluated culture media. RPFA and STX had good correlation indices between the total and typical colony counts as well as with coagulase and the thermonuclease-positive colony counts. Thus, there is a better association between coagulase and thermonuclease production to typical colony morphology developed on these culture media, leading to more accurate and reliable results than with BP, which demonstrated lower correlation indices between these counts. 2009 Elsevier Ltd. All rights reserved.
This dissertation is concerned with the syntactic and semantic status of negative indefinites in Afrikaans. The problem posed by negative indefinites is that their interpretation appears to differ across double negation (DN) and negative concord (NC) languages. With respect to negative indefinites, Afrikaans displays features that distinguish it from both typical NC and typical DN languages. Contrary to most NC languages, and similarly to DN languages, standard Afrikaans does not allow negati...
Rosemari Laura Cardoso
O objetivo do presente estudo foi avaliar o potencial antimicrobiano do extrato de própolis, substância natural conhecida pelas suas propriedades terapêuticas, frente a 67 isolados microbianos da otite canina. A Concentração Bactericida Mínima (CBM), do extrato de própolis, frente a isolados de Staphylococcus coagulase positiva e a Concentração Fungicida Mínima (CFM) para isolados de Malassezia pachydermatis foram determinadas utilizando-se a técnica de microdiluição em caldo. A CBM foi...
Matsunaga, T; Yoshida, T; Kamata, S; Uchida, K
Strains of Staphylococcus species isolated from bovine mastitic milk at 66 dairy farms in Japan during the period from November 1988 to May 1989 were identified, and examined for their drug susceptibility and beta-lactamase production in order to clarify an epidemiological aspect of bovine mastitis caused by staphylococci. The results of bacteriological identification showed that the most predominant species was S. xylosus. Other major species isolated were S. aureus, S. sciuri and S. hyicus. Thirty-eight (71.7%) isolates of S. xylosus, 21 (45.7%) of S. aureus and 5 (71.4%) of S. epidermidis were positive for beta-lactamase production. Most of the beta-lactamase-producers of S. aureus were classified as high producers, although all of the beta-lactamase-positive S. xylosus isolates remained to be low producers. All isolates of S. aureus were sensitive to methicillin and cloxacillin at 6.25 micrograms/ml and 1.56 micrograms/ml, respectively, and none of methicillin-resistant S. aureus were detected. Isolates of other species were considered to be susceptible to 6 beta-lactams, in contrast to human isolates, but antibacterial activities of penicillin G and ampicillin were affected more strongly by beta-lactamase than those of methicillin, cloxacillin, cefazolin and cefoperazone.
Investigations on the efficacy of routinely used phenotypic methods compared to genotypic approaches for the identification of staphylococcal species isolated from companion animals in Irish veterinary hospitals
Background Identification of Staphylococci to species level in veterinary microbiology is important to inform therapeutic intervention and management. We report on the efficacy of three routinely used commercial phenotypic methods for staphylococcal species identification, namely API Staph 32 (bioMérieux), RapID (Remel) and Staph-Zym (Rosco Diagnostica) compared to genotyping as a reference method to identify 52 staphylococcal clinical isolates (23 coagulase positive; 29 coagulase negative) from companion animals in Irish veterinary hospitals. Results Genotyping of a 412 bp fragment of the staphylococcal tuf gene and coagulase testing were carried out on all 52 veterinary samples along with 7 reference strains. In addition, genotyping of the staphylococcal rpoB gene, as well as PCR-RFLP of the pta gene, were performed to definitively identify members of the Staphylococcus intermedius group (SIG). The API Staph 32 correctly identified all S. aureus isolates (11/11), 83% (10/12) of the SIG species, and 66% (19/29) of the coagulase negative species. RapID and Staph-Zym correctly identified 61% (14/23) and 0% (0/23) respectively of the coagulase-positives, and 10% (3/29) and 3% (1/29) respectively of the coagulase-negative species. Conclusions Commercially available phenotypic species identification tests are inadequate for the correct identification of both coagulase negative and coagulase positive staphylococcal species from companion animals. Genotyping using the tuf gene sequence is superior to phenotyping for identification of staphylococcal species of animal origin. However, use of PCR-RFLP of pta gene or rpoB sequencing is recommended as a confirmatory method for discriminating between SIG isolates. PMID:23635328
da Silva, Sabina Dos Santos Paulino; Cidral, Thiago André; Soares, Maria José dos Santos; de Melo, Maria Celeste Nunes
Food handlers carrying enterotoxin-producing Staphylococcus are a potential source of food poisoning. The aim of this study was to analyze genes encoding enterotoxins in coagulase-positive Staphylococcus (CoPS) and coagulase-negative Staphylococcus (CoNS) isolated from the anterior nostrils and hands of food handlers at a university restaurant in the city of Natal, Northeast Brazil. Thirty food handlers were screened for the study. The isolates were subjected to Gram staining, a bacitracin sensitivity test, mannitol fermentation, and catalase and coagulase tests. CoNS and CoPS strains were subsequently identified by a Vitek 2 System (BioMerieux, France) and various biochemical tests. Polymerase chain reaction was used to detect genes for enterotoxins A, B, C, D, E, G, H, and I (sea, seb, sec, sed, see, seg, seh, and sei) and a disc-diffusion method was used to determine susceptibility to several classes of antimicrobials. All food handlers presented staphylococci on their hands and/or noses. The study found 58 Staphylococcus spp., of which 20.7% were CoPS and 79.3% were CoNS. S. epidermidis was the most prevalent species. Twenty-nine staphylococci (50%) were positive for one or more enterotoxin genes, and the most prevalent genes were seg and sei, each with a frequency of 29.3%. Indeed, CoNS encoded a high percentage of enterotoxin genes (43.5%). However, S. aureus encoded even more enterotoxin genes (75%). Most isolates showed sensitivity to the antibiotics used for testing, except for penicillin (only 35% sensitive). The results from this study reinforce that coagulase-negative as well as coagulase-positive staphylococci isolated from food handlers are capable of genotypic enterotoxigenicity.
Ford, L. [Tufts Univ., Medford, MA (United States); Roman, Th. [Connecticut Univ., Farmington, CT (United States)
We could travel at light velocity or more if we mastered a new form of energy called negative energy. This energy is predicted by the quantum theory and follows from the Heisenberg principle applied to the void. In the void the density of energy is null in average but in fact fluctuates around zero according to the quantum theory, so we can find places with negative energy. This concept has nothing to do with antimatter or with the energy introduced in the recent theories of anti-gravity. According to the theory of general relativity the presence of matter and energy distorts the space-time geometry. In the presence of negative energy strange phenomena might happen. A tunnel could bridge 2 distant points in the universe, and inside it travelling at light velocity or more would be possible. (A.C.)
Negative Polarity Items (NPIs) are words or expressions that exhibit a restricted distribution to certain negative contexts only. For example, yet is an NPI and must appear in the scope of a negation: Mary has *(not) finished yet. The existence of NPIs such as yet gives rise to a learnability
Carricajo, Anne; Treny, Axel; Fonsale, Nathalie; Bes, Michele; Reverdy, Marie Elisabeth; Gille, Yves; Aubert, Gerald; Freydiere, Anne Marie
CHROMagar Staph aureus (CSAM) (CHROMagar Microbiology, Paris, France) is a new chromogenic medium designed to enable detection of colonies of Staphylococcus aureus by their pink color. A total of 775 specimens were cultured in parallel on CHROMagar Staph aureus and conventional media. Among the 267 S. aureus strains recovered on at least one medium, 263 were isolated on CSAM medium (sensitivity, 98.5%), and 245 (sensitivity, 91.8%) were isolated on conventional media. The specificity of presumptive identification of S. aureus on the basis of pink colony color on CSAM medium was 97% (493 of 508). This specificity increased to 100% when coagulase detection with the Staphychrom coagulase test was added and to 98.8% when S. aureus surface components were detected by agglutination in the Pastorex Staph Plus test. Susceptibility testing of 67 S. aureus strains, performed in parallel on pink CSAM colonies and on colonies grown on blood agar, gave similar results. Thus, rapid and accurate recognition and identification of S. aureus isolates were achieved with CSAM as the primary isolation medium, followed by the staphylocoagulase Staphychrom test. Antimicrobial susceptibility testing (disk-diffusion method or ATB STAPH System) can be performed directly on pink CSAM colonies. PMID:11427572
Papa, Rosanna; Selan, Laura; Parrilli, Ermenegilda; Tilotta, Marco; Sannino, Filomena; Feller, Georges; Tutino, Maria L; Artini, Marco
Microbial biofilms have great negative impacts on the world's economy and pose serious problems to industry, public health and medicine. The interest in the development of new approaches for the prevention and treatment of bacterial adhesion and biofilm formation has increased. Since, bacterial pathogens living in biofilm induce persistent chronic infections due to the resistance to antibiotics and host immune system. A viable approach should target adhesive properties without affecting bacterial vitality in order to avoid the appearance of resistant mutants. Many bacteria secrete anti-biofilm molecules that function in regulating biofilm architecture or mediating the release of cells from it during the dispersal stage of biofilm life cycle. Cold-adapted marine bacteria represent an untapped reservoir of biodiversity able to synthesize a broad range of bioactive compounds, including anti-biofilm molecules. The anti-biofilm activity of cell-free supernatants derived from sessile and planktonic cultures of cold-adapted bacteria belonging to Pseudoalteromonas, Psychrobacter, and Psychromonas species were tested against Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa strains. Reported results demonstrate that we have selected supernatants, from cold-adapted marine bacteria, containing non-biocidal agents able to destabilize biofilm matrix of all tested pathogens without killing cells. A preliminary physico-chemical characterization of supernatants was also performed, and these analyses highlighted the presence of molecules of different nature that act by inhibiting biofilm formation. Some of them are also able to impair the initial attachment of the bacterial cells to the surface, thus likely containing molecules acting as anti-biofilm surfactant molecules. The described ability of cold-adapted bacteria to produce effective anti-biofilm molecules paves the way to further characterization of the most promising molecules and to test their
Ding, Qun; Li, Dai-qing; Wang, Peng-hua; Chu, Yue-jie; Meng, Shu-you; Sun, Qian
To examine the distribution patterns of pathogens isolated from the patients with diabetic foot ulcers and explore the risk factors for infections of methicillin-resistant S. aureus (MRSA) or methicillin-resistant S. epidermidis (MRSE). A total of 388 diabetic-foot patients hospitalized at Tianjin Metabolic Diseases Hospital between January 2008 and June 2010 were recruited. The distribution profiles of pathogens isolated from diabetic foot ulcers were summarized. The patients with S. aureus infections were divided into MRSA and MSSA groups while those with S. epidermidis infections into MRSE and MSSE groups. The clinical features of these patients were compared between all groups. Logistic regression was employed to identify the risk factors for the MRSA/MRSE infections. A total of 362 pathogens were isolated from them. And the Gram-positive bacteria were the most predominant (57.2%, 207/362), followed by Gram-negative bacilli (39.2%, 142/362) and true fungi (3.6%, 13/362). The three most frequently isolated pathogens were S. aureus (27.1%), S. epidermidis (18.8%) and Pseudomonas aeruginosa (15.5%). Statistically significant differences existed in antibiotic usage in 6 months prior to hospitalization, course of ulcer, ulcer size, deep ulcer, osteomyelitis, hypertension, anemia, hypoproteinemia and erythrocyte sedimentation rate between the patients infected with MRSA and MSSA (P infection was correlated with recurrent ulcer, osteomyelitis, hypoproteinemia, HbA1c and lower total serum protein (P risk factors for the MRSA infection. And HbA1c was a risk factor for the MRSE infection. In the present study, the Gram-positive cocci are the main pathogens isolated from diabetic foot ulcers. And S. aureus and S. epidermidis are the most frequently isolated pathogens. Antibiotic usage in 6 months prior to hospitalization, long course of ulcer, osteomyelitis, hypertension and hypoproteinemia are risk factors for the MRSA infection. And HbA1c is a risk factor for the MRSE
Full Text Available Microbial biofilms have great negative impacts on the world’s economy and pose serious problems to industry, public health and medicine. The interest in the development of new approaches for the prevention and treatment of bacterial adhesion and biofilm formation has increased. Since, bacterial pathogens living in biofilm induce persistent chronic infections due to the resistance to antibiotics and host immune system. A viable approach should target adhesive properties without affecting bacterial vitality in order to avoid the appearance of resistant mutants. Many bacteria secrete anti-biofilm molecules that function in regulating biofilm architecture or mediating the release of cells from it during the dispersal stage of biofilm life cycle. Cold-adapted marine bacteria represent an untapped reservoir of biodiversity able to synthesize a broad range of bioactive compounds, including anti-biofilm molecules.The anti-biofilm activity of cell-free supernatants derived from sessile and planktonic cultures of cold-adapted bacteria belonging to Pseudoalteromonas, Psychrobacter and Psychromonas species were tested against Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa strains. Reported results demonstrate that we have selected supernatants, from cold-adapted marine bacteria, containing non-biocidal agents able to destabilize biofilm matrix of all tested pathogens without killing cells. A preliminary physico-chemical characterization of supernatants was also performed, and these analyses highlighted the presence of molecules of different nature that act by inhibiting biofilm formation. Some of them are also able to impair the initial attachment of the bacterial cells to the surface, thus likely containing molecules acting as anti-biofilm surfactant molecules.The described ability of cold-adapted bacteria to produce effective anti-biofilm molecules paves the way to further characterization of the most promising molecules
Munier, Anne-Lise; de Lastours, Victoire; Barbier, François; Chau, Françoise; Fantin, Bruno; Ruimy, Raymond
Fluoroquinolone-resistant staphylococci (FQRS) are primarily selected in the nasal microbiota during fluoroquinolone (FQ) treatment. To gain insight into the dynamics of the emergence of FQRS, 49 hospitalised patients (HPs) and 62 community patients (CPs) treated with FQs were studied. Nasal swabs were collected before (T0), at the end of (T1) and 1 month after (T2) FQ treatment. FQRS were identified by mass spectrometry. Antibiotic resistance was determined. Pre- and post-exposure staphylococci populations were compared phenotypically and by MLST to determine the origin of FQRS. At T0, 33/49 HPs (67%) and 24/62 CPs (39%) carried FQRS (OR=3.3, 95% CI: 1.4-7.9; P<0.001). Among patients with no FQRS at T0, 15/16 HPs (94%) and 16/38 CPs (42%) had FQRS detected at T1 and/or T2 (OR=19.6, 95% CI: 2.5-902; P<0.001). Among FQRS having emerged, co-resistance to meticillin was detected in 87% and 82% of HPs and CPs, respectively. No selection of resistance emerging from the initial microbiota was evidenced. FQRS showed decreased species diversity in favour of Staphylococcus haemolyticus and Staphylococcus epidermidis. As a consequence of FQ treatment, acquisition of FQRS in the nasal microbiota is frequent in the community and almost inevitable in hospitals. Acquisition from extranasal sites prevails. A restriction in species diversity in favour of more pathogenic and resistant species occurs. This highlights the major impact of FQ treatment on nasal microbiota, the role of the ecological environment in the emergence of FQRS, and the high-risk of dissemination of resistant staphylococci. Copyright © 2015 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.
Negative ions tend to stratify in electronegative plasmas with hot electrons (electron temperature Te much larger than ion temperature Ti, Te > Ti ). The boundary separating a plasma containing negative ions, and a plasma, without negative ions, is usually thin, so that the negative ion density falls rapidly to zero-forming a negative ion density front. We review theoretical, experimental and numerical results giving the spatio-temporal evolution of negative ion density fronts during plasma ignition, the steady state, and extinction (afterglow). During plasma ignition, negative ion fronts are the result of the break of smooth plasma density profiles during nonlinear convection. In a steady-state plasma, the fronts are boundary layers with steepening of ion density profiles due to nonlinear convection also. But during plasma extinction, the ion fronts are of a completely different nature. Negative ions diffuse freely in the plasma core (no convection), whereas the negative ion front propagates towards the chamber walls with a nearly constant velocity. The concept of fronts turns out to be very effective in analysis of plasma density profile evolution in strongly non-isothermal plasmas.
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
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.
SUSCEPTIBILITIES/RESISTANCE OF Staphylococcus coagulase POSITIVE AND Streptococcus sp. ISOLATED FROM THE MILK OF COWS PRESENTING CLINICAL MASTITIS IN MILK BASIN OF GOIÂNIA RESISTÊNCIA DE Staphylococcus coagulase POSITIVA E Streptococcus sp. ISOLADOS DO LEITE DE VACAS COM MASTITE CLÍNICA NA BACIA LEITEIRA DE GOIÂNIA
Luiz Antônio Franco da Silva
Full Text Available
It was verified the spectrum of susceptibilities/resistance of 76 samples of Staphylococcus coagulase positive and 51 samples of Streptococcus sp., isolated from milk of 231 cows presenting clinical mastitis. The diffusion method was used in plate of Kirby-Bauer, being tested ten active principles used in cases of clinical mastitis. Natural penicillin, chloranphenicol, tetraciclyne, kanamicyn, gentamicyn, nitrofurantoin, trimetopryn + sulfametoxazol, enrofloxacyn, perlimicyn and ceftiofur were tested. The samples of Staphylococcus coagulase positive presented a spectrum of larger resistance for the penicillin (78.9%, followed by trimetoprim + sulfametoxazol (59.2% and nitrofurantoin (57.8%. The largest susceptibility frequency was found in enrofloxacyn (96%, in perlimicyn (94% and in ceftiofur (94%. For the samples of Streptococcus sp. there was a profile resistance for penicillin (92%, chloranphenicol (74.5% and trimetoprim + sulfametoxazol (52%. A sensibility profile was also verified for enrofloxacyn (96%, ceftiofur (92% and perlimicyn (92%.
KEY-WORDS: Bovine mastitis; resistance; susceptibilities.
Verificou-se o espectro de sensibilidade/resistência de 76 cepas de Staphylococcus coagulase positiva e 51 cepas de Streptococcus sp., isolados do leite proveniente de 231 vacas que apresentaram mastite clínica. Utilizou-se o método de difusão em placa, segundo Kirby-Bauer, testando-se 10 princípios ativos: penicilina natural, cloranfenicol, tetraciclina, kanamicina, gentamicina, nitrofurantoína, trimetoprim + sulfametoxazol, enrofloxacina, perlimicina e ceftiofur. As cepas de Staphylococcus coagulase positiva apresentaram um espectro de resistência maior para a penicilina (78,9%, vindo a seguir trimetoprim + sulfametoxazol (59,2% e
Miwa, N; Kawamura, A; Masuda, T; Akiyama, M
An outbreak of staphylococcal food poisoning due to an egg yolk (EY) reaction-negative strain occurred in Japan. Twenty-one of 53 dam construction workers who ate boxed lunches prepared at their company cafeteria became ill, and eight required hospital treatment. The outbreak showed a typical incubation time (1.5-4 h with a median time of 2.7 h) and symptoms (vomiting and diarrhea) of staphylococcal food poisoning. Staphylococcus aureus, which produces staphylococcal enterotoxin (SE) A, was isolated from four fecal specimens of eight patients tested. Scrambled egg in the boxed lunches contained 20-40 ng/g of SEA, and 3.0 x 10(9)/g of viable S. aureus cells that produced this toxin. All isolates from patients and the food were EY reaction-negative, coagulase type II, and showed the same restriction fragment length polymorphism (RFLP) pattern. We concluded that the outbreak was caused by scrambled egg contaminated with EY reaction-negative S. aureus. In Japan, outbreaks of staphylococcal food poisoning are mainly caused by EY reaction-positive S. aureus, and EY reaction-negative colonies grown on agar plates containing EY are usually not analyzed further for detection of S. aureus. The present outbreak suggested that EY reaction-negative isolates should be subjected to further analysis to detect the causative agents of staphylococcal food poisoning.
Rosangela E. Ziech
Full Text Available O objetivo do presente trabalho foi investigar o potencial antimicrobiano do oleorresina de Copaifera reticulata Ducke em isolados de Staphylococcus coagulase positiva (SCP provenientes de casos de otite externa em cães. O método de microdiluição em caldo foi utilizado para determinação da concentração inibitória mínima (CIM e concentração bactericida mínima (CBM de oleorresina de copaíba. Em adição, foi determinado o perfil de suscetibilidade aos antimicrobianos dos isolados de SCP pelo método de difusão em ágar. Oito classes de antimicrobianos foram usadas para o cálculo de multirresistência antimicrobiana. A determinação da composição química do oleorresina de copaíba foi realizada por cromatografia em fase gasosa acoplada à espectrometria de massas (GC/MS, sendo que β-cariofileno, β-bisaboleno e (E-α-bergamoteno foram os compostos majoritários. O oleorresina de copaíba demonstrou CIM90 de 0,164mg/mL e CBM90 de 1,31mg/mL. A multirresistência foi verificada em 27% das cepas testadas. Os resultados sugerem que o oleorresina de copaíba exerceu atividade bacteriostática e bactericida mesmo em cepas multirresistentes de Staphylococcus coagulase-positiva.
Full Text Available Chronic bone and joint infections (BJI are devastating diseases. Relapses are frequently observed, as some pathogens, especially staphylococci, can persist intracellularly by expressing a particular phenotype called small colony variant (SCV. As natural killer (NK cells are lymphocytes specialized in the killing of host cells infected by intracellular pathogens, we studied NK cells of patients with chronic BJI due to staphylococci expressing or not SCVs (10 patients in both groups. Controls were patients infected with other bacteria without detectable expression of SCVs, and healthy volunteers. NK cell phenotype was evaluated from PBMCs by flow cytometry. Degranulation capacity was evaluated after stimulation with K562 cells in vitro. We found that NK cells were activated in terms of CD69 expression, loss of CD16 and perforin, in all infected patients in comparison with healthy volunteers, independently of the SCV phenotype. Peripheral NK cells in patients with chronic BJI display signs of recent activation and degranulation in vivo in response to CD16-mediated signals, regardless of the type of bacteria involved. This could involve a universal capacity of isolates responsible for chronic BJI to produce undetectable SCVs in vivo, which might be a target of future intervention.
Gu, Jingmin; Lu, Rong; Liu, Xiaohe; Han, Wenyu; Lei, Liancheng; Gao, Yu; Zhao, Honglei; Li, Yue; Diao, Yuwen
LysGH15, a phage endolysin, exhibits a particularly broad lytic spectrum against Staphylococcus aureus, especially methicillin-resistant S. aureus (MRSA). Sequence analysis reveals that this endolysin contains a C-terminal cell wall binding domain (SH3b), which causes the endolysin to bind to host strains. In this study, the substrate binding affinity of the SH3b domain (LysGH15B) was evaluated. A fusion protein of LysGH15B and green fluorescent protein (LysGH15B-GFP) were cloned and expressed in Escherichia coli. Laser scanning confocal microscopy was used to detect the fluorescence of the treated cells irradiated at different excitation wavelengths and to determine the binding activity of LysGH15B-GFP and GFP. We found that LysGH15B-GFP not only generated green fluorescence, but, more importantly, also displayed specific affinity to staphylococcal isolates, especially MRSA. In contrast, the single GFP did not display any binding activity. The high affinity was attributed to the portion of LysGH15B and the binding activity of the fusion protein was specific to staphylococci. This study provides an insight into the SH3b domain of LysGH15. The specific binding activity may cause LysGH15B to serve as an anchoring device, and offer an alternative approach for cell surface attachment onto staphylococci.
Full Text Available This paper presents the application of simple, and therefore cheap, planar resonators for building 3D isotropic metamaterials. These resonators are: a broadside-coupled split ring resonator with a magnetic response providing negative permeability; an electric dipole terminated by a loop inductor together with a double H-shaped resonator with an electric response providing negative permittivity. Two kinds of 3D isotropic single negative metamaterials are reported. The first material consists of unit cells in the form of a cube bearing on its faces six equal planar resonators with tetrahedral symmetry. In the second material, the planar resonators boxed into spherical plastic shells and randomly distributed in a hosting material compose a real 3D volumetric metamaterial with an isotropic response. In both cases the metamaterial shows negative permittivity or permeability, according to the type of resonators that are used. The experiments prove the isotropic behavior of the cells and of the metamaterial specimens.
... life-threatening illness. Prevention Prompt treatment of related infections may reduce the risk of meningitis. Alternative Names Gram-negative meningitis Images Central nervous system and peripheral nervous ...
Holt, MS; Andersen, SS; Andersen, Ove
patients with only local symptoms of infection. The microorganisms isolated from the tips of the epidural catheters were coagulase-negative staphylococci (41%), Staphylococcus aureus (35%), Gram-negative bacilli (14%) and others (10%). The Gram-negative bacilli and S. aureus caused serious infections more...... frequently than the others. We discuss the symptoms and diagnosis of spinal epidural abscess and suggest a proposal for prophylactic and diagnostic guidelines for epidural catheter-related infections. Comment in: J Hosp Infect. 1997 Mar;35(3):245....
Holt, H M; Gahrn-Hansen, B; Andersen, S S
coagulase- negative staphylococci (41%), Staphylococcus aureus (35%), Gram-negative bacilli (14%) and other bacteria (10%). The Gram-negative bacilli and S. aureus caused serious infections more frequently than the others. We discuss the symptoms and diagnosis of spinal epidural abscess and propose......Seventy-eight patients with culture-positive epidural catheters were studied. Fifty-nine had symptoms of exit site infection and 11 patients had clinical meningitis, two of whom also had en epidural abscess. This corresponds to a local infection incidence of at least 4.3% and an incidence...
VOGES-PROSKAUER AND COAGULASE TESTS TO THE LABORATORIAL DIAGNOSTIC OF Staphylococcus aureus INCLUDED IN THE EPIDEMIOLOGY OF BOVINE MASTITIS UTILIZAÇÃO DO TESTE DE VOGES-PROSKAUER E DA COAGULASE PARA O DIAGNÓSTICO LABORATORIAL DE Staphylococcus aureus ENVOLVIDOS NA EPIDEMIOLOGIA DA MASTITE BOVINA
Luiz Francisco Zafalon
Full Text Available Diagnostic characteristics of acetoin production to the Staphylococcus aureus isolated from milk of mammary quarters of cows with mastitis, orifices of teats and mouthpieces of the liners were studied, in agreement to different coagulation grades presented by strains in the tube coagulase test with rabbit plasma and different rain levels in the environment of the herd. The production of acetoin and the coagulase test were performed after growth on blood agar base supplemented with sheep blood, Gram staining and catalase production. All strains were confirmed by molecular identification at species level by species-specific PCR. Among 329 samples identified, 167 were isolated from milk, 117 from teat orifices and 45 from mouthpieces of the liners. Among these samples, 85.0%, 71.1% and 69.2% strains originated from the milk, mouthpieces and orifices, respectively, presented whole coagulation in the coagulase test. Reduced specificities were obtained to the acetoin production to different coagulation grades on the few rains period as well as on the many rains period. Sensitivities to the acetoin production were higher on the few rains period to all sources and coagulation grades, except to strains from orifices with incomplete coagulation and strains from mouthpieces with complete coagulation. It is recommended the acetoin production test how a good diagnostic strategy to the S. aureus identification, combined with other tests how coagulase, mainly in the milk samples originated from mastitic mammary quarters and on periods with little occurrence of rains.
Enterotoxin Genes 148 B. Membrane -Bound Toxin Precursor 148 C. Plasmid Involvement in Enterotoxin B Production 149 D. Genetic Analysis of Enterotoxin A...1985) extended this observation to SEB. They were able to completely carboxymethylate its six histidine residues and to demonstrate a loss of...B. Membrane -Bound Toxin Precursor Blobel and Dobberstein (1975) proposed a model system for protein secre- tion involving an additional sequence of
Dedy Subandowo -
Full Text Available Abstract: This research entitled "Negation Affixes in English". This study is aimed to describe the various negation affixes in English, morphological process, morphophonemic and meaning. The research data were taken from various sources of English grammar book, morphology, research journal and the book which relatees to the research. English grammar books used in this study are written by Otto Jesperson, Marcella Frank, Greenbaum and Geoffrey Leech. The method used in this research is the descriptive-qualitative method. While the data collection techniques are performed by using jot-down method. And the results of analysis are presented in tabular form and descriptive method. The result of the research shows that English has six types of negative affixes which are categorized by the intensity of its appearance, such as dis-, in-, non-, un-, anti- and -less. Based on the function, negation affixes are divided into several categories such as adjectives, nouns, verbs, and adverbs. The morphophonemic affix in- has four allomorphs, they are in-, im-, il- and ir- . While the analysis revealed that negation affixes have some basic meanings, such as ‘not’, ‘without’, and ‘anti’.
Full Text Available Introduction. One of the main ways to control microorganisms’ resistance to antibiotics is to find substances that are able to overcome it and potentiate antibiotics action, in particular to neutralize the antibiotic-inactivating enzymes or block the active efflux of antibiotic from microbial cells. Every year there is a growing interest in the therapeutic potential of herbal active compounds as modifiers of antibiotic resistance including MLS-resistance (macrolide-lincosamide-streptoramin B. It should be emphasized that a number of biologically active substances of plant origin can potentiate antimicrobial activity of erythromycin (ERY against MLS-resistant staphylococci. The present study was designed to investigate the antibacterial and synergistic effects of eight Ukrainian ethanol medicinal plant extracts with erythromycin against skin strains of staphylococci with inducible phenotype of MLS-resistance. Material & methods. S. aureus and S. epidermidis strains were tested for susceptibility to antibiotics of MLS-group by disk diffusion test. Effective antimicrobial concentrations of plant extracts and erythromycin were determined by two-fold serial dilution in nutrient agar and broth. Combinatory effects between organic extracts and ERY were assessed using the checkerboard assay against tested strains to evaluate culture growth in the presence of two antimicrobials with different concentrations. Results & discussion. The Alnus incana L. fruits extract was the most potent inhibitor against tested strains (MIC 40.625-162.5 µg/mL; while Geranium pratense L. rhizomes extract exhibited the least antimicrobial activity (MIC 650-2,600 µg/mL. The Alnus incana L. fruits extract and the Geranium pratense L. rhizomes extract showed synergistic effect with erythromycin against 100% strains of staphylococci (average FICI 0.028 – 0.057; p<0.001. In the presence of 1/4 MIC of ERY Alnus incana L. fruits extract antimicrobial concentration was
Skovsgaard, Morten; Søberg, Pernille Frantz
I adskillige år er det blevet debatteret, hvorvidt nyhedernes negative fokus har konsekvenser for borgerne, og om det i sid-ste ende får flere til at vende ryggen til nyhederne. Vores viden om effekterne af positive og negative nyheder er dog begrænset, og derfor undersøges det i denne artikel......, hvordan henholdsvis positive og negative tv-nyheder påvirker seernes humør, hukom-melse af information fra indslaget og lyst til at se yderligere tv-nyheder. Det gør vi i et survey-eksperiment (N=204), hvor tre grupper så enten et originalt indslag eller det samme indslag klippet med henholdsvis et...
Mulligan, Neil W.; Peterson, Daniel J.
A fundamental property of human memory is that repetition enhances memory. Peterson and Mulligan (2012) recently documented a surprising "negative repetition effect," in which participants who studied a list of cue-target pairs twice recalled fewer targets than a group who studied the pairs only once. Words within a pair rhymed, and…
Artiklen formidler resultater fra en longitudinel undersøgelse af det selvforstærkende, negative samspil imellem udvikling og vedligeholdelse af alderdomsdepression via primære miljøbelastninger og via den deprimerede ældre som belastning for miljøet, som i sin tur "svarer negativt" på lidelsen og...
Gelasakis, A I; Angelidis, A S; Giannakou, R; Filioussis, G; Kalamaki, M S; Arsenos, G
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
Full Text Available Staphylococcus schleiferi subsp. schleiferi is a coagulase-negative Staphylococcus which has been described as a pathogen responsible for various nosocomial infections including bacteremia, brain abscess, and infection of intravenous pacemakers. Recently, such bacteria have been described to be found typically on skin and mucosal surfaces. It is also believed to be a part of the preaxillary human flora and more frequently found in men. It is very similar in its pathogenicity with Staphylococcus aureus group and expresses a fibronectin binding protein. Literature on this pathogen reveals that it commonly causes otitis among dogs because of its location in the auditory meatus of canines. Also, it has strong association with pyoderma in dogs. The prime concern with this organism is the antibiotic resistance and relapse even after appropriate treatment. Very rarely, if any, cases have been reported about prostatic abscess (PA with this microbe. Our patient had a history of recurrent UTIs and subsequent PA resulting in S. schleiferi bacteremia in contrast to gram negative bacteremia commonly associated with UTI. This organism was found to be resistant to methicillin, in spite of being negative for PBP2, which is a rare phenomenon and needs further studies.
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.
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.
Suriyasathaporn, W.; Chupia, V.; Sing-Lah, T.; Wongsawan, K.; Mektrirat, R.; Chaisri, W.
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. PMID:25049697
Full Text Available Os estafilococos coagulase-negativos (ECN são importantes agentes etiológicos das bacteremias hospitalares e freqüentemente considerados como contaminantes de hemoculturas. No período de outubro de 1990 a setembro de 1992, foram estudadas 300 hemoculturas positivas para ECN no Hospital São Paulo, sendo 141 bacteremias consideradas de origem hospitalar. Com o objetivo de diferenciar as bacteremias hospitalares verdadeiras das contaminantes por ECN, foram definidos critérios clínicos e microbiológicos. Apenas 20,6% das bacteremias hospitalares por ECN foram consideradas como verdadeiras. A maior freqüência de recém-nascidos internados na unidade de terapia intensiva neonatal, a presença de cateter intravascular e a utilização de nutrição parenteral foram achados significativos. Não houve diferença significante quanto a resistência a oxacilina e produção de SLIME entre os ECN isolados das bacteremias verdadeiras e contaminantes. O critério clínico e a positividade da hemocultura até 48 horas após a incubação, utilizados em nossa definição, foram úteis para caracterizar as bacteremias verdadeiras por ECN.
Human Resources Department
Members of the personnel entitled to annual leave and, where appropriate, saved leave and/or compensatory leave are requested to take note of the new arrangements described below, which were recommended by the Standing Concertation Committee (SCC) at its meeting on 1Â September 2005 and subsequently approved by the Director-General. The changes do not apply to members of the personnel participating in the Progressive Retirement Programme (PRP) or the Part-time Work as a pre-retirement measure, for whom the specific provisions communicated at the time of joining will continue to apply. Â Negative balances in annual leave, saved leave and/or compensatory leave accounts at the end of the leave year (30th September) and on the date on which bonuses are credited to the saved leave account (31st December): Where members of the personnel have a leave account with a negative balance on 30Â September and/or 31Â December, leave will automatically be transferred from one account to another on the relevant dates i...
Human Resources Department
Members of the personnel entitled to annual leave and, where appropriate, saved leave and/or compensatory leave are requested to take note of the new arrangements described below, which were recommended by the Standing Concertation Committee (SCC) at its meeting on 1 September 2005 and subsequently approved by the Director-General. The changes do not apply to members of the personnel participating in the Progressive Retirement Programme (PRP) or the Part-time Work as a pre-retirement measure, for whom the specific provisions communicated at the time of joining will continue to apply. Negative balances in annual leave, saved leave and/or compensatory leave accounts at the end of the leave year (30th September) and on the date on which bonuses are credited to the saved leave account (31st December): Where members of the personnel have a leave account with a negative balance on 30 September and/or 31 December, leave will automatically be transferred from one account to another on the relevant dates in or...
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.
Lavenda, B. H.
A modification of the second law is required for a system with a bounded density of states and not the introduction of a `negative' temperature scale. The ascending and descending branches of the entropy versus energy curve describe particle and hole states, having thermal equations of state that are given by the Fermi and logistic distributions, respectively. Conservation of energy requires isentropic states to be isothermal. The effect of adiabatically reversing the field is entirely mechanical because the only difference between the two states is their energies. The laws of large and small numbers, leading to the normal and Poisson approximations, characterize statistically the states of infinite and zero temperatures, respectively. Since the heat capacity also vanishes in the state of maximum disorder, the third law can be generalized in systems with a bounded density of states: the entropy tends to a constant as the temperature tends to either zero or infinity.
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
Full Text Available Obesity promotes excessive inflammation, which is associated with senescence-like changes in visceral adipose tissue (VAT and the development of type 2 diabetes (T2DM and cardiovascular diseases. We have reported that a unique population of CD44hi CD62Llo CD4+ T cells that constitutively express PD-1 and CD153 exhibit cellular senescence and cause VAT inflammation by producing large amounts of osteopontin. Weight loss improves glycemic control and reduces cardiovascular disease risk factors, but its long-term effects on cardiovascular events and longevity in obese individuals with T2DM are somewhat disappointing and not well understood. High-fat diet (HFD-fed obese mice were subjected to weight reduction through a switch to a control diet. They lost body weight and visceral fat mass, reaching the same levels as lean mice fed a control diet. However, the VAT of weight reduction mice exhibited denser infiltration of macrophages, which formed more crown-like structures compared to the VAT of obese mice kept on the HFD. Mechanistically, CD153+ PD-1+ CD4+ T cells are long-lived and not easily eliminated, even after weight reduction. Their continued presence maintains a self-sustaining chronic inflammatory loop via production of large amounts of osteopontin. Thus, we concluded that T-cell senescence is essentially a negative legacy effect of obesity.
Full Text Available Abstract Background Bloodstream infections are associated with significant patient morbidity and mortality. Antimicrobial susceptibility patterns should guide the choice of empiric antimicrobial regimens for patients with bacteremia. Methods From January to December of 2002, 82,569 bacterial blood culture isolates were reported to The Surveillance Network (TSN Database-USA by 268 laboratories. Susceptibility to relevant antibiotic compounds was analyzed using National Committee for Clinical Laboratory Standards guidelines. Results Coagulase-negative staphylococci (42.0%, Staphylococcus aureus (16.5%, Enterococcus faecalis (8.3%, Escherichia coli (7.2%, Klebsiella pneumoniae (3.6%, and Enterococcus faecium (3.5% were the most frequently isolated bacteria from blood cultures, collectively accounting for >80% of isolates. In vitro susceptibility to expanded-spectrum β-lactams such as ceftriaxone were high for oxacillin-susceptible coagulase-negative staphylococci (98.7%, oxacillin-susceptible S. aureus (99.8%, E. coli (97.3%, K. pneumoniae (93.3%, and Streptococcus pneumoniae (97.2%. Susceptibilities to fluoroquinolones were variable for K. pneumoniae (90.3–91.4%, E. coli (86.0–86.7%, oxacillin-susceptible S. aureus (84.0–89.4%, oxacillin-susceptible coagulase-negative staphylococci (72.7–82.7%, E. faecalis (52.1%, and E. faecium (11.3%. Combinations of antimicrobials are often prescribed as empiric therapy for bacteremia. Susceptibilities of all blood culture isolates to one or both agents in combinations of ceftriaxone, ceftazdime, cefepime, piperacillin-tazobactam or ciprofloxacin plus gentamicin were consistent (range, 74.8–76.3% but lower than similar β-lactam or ciprofloxacin combinations with vancomycin (range, 93.5–96.6%. Conclusion Ongoing surveillance for antimicrobial susceptibility remains essential, and will enhance efforts to identify resistance and attempt to limit its spread.
Mário Sérgio Azevedo Resta
Full Text Available A multiplicacão de Staphylococcus aureus e a produção de enterotoxinas estafilocócicas dependem de fatores intrínsecos e da temperatura de armazenamento dos alimentos. O objetivo deste trabalho foi discutir o padrão estafilococos coagulase positiva (ECP para massas alimentícias, estabelecido pela RDC n.º 12 de 2001 da ANVISA, baseando-se nos resultados de atividade de água (a w, potencial hidrogeniônico (pH e contagem de ECP. Cinquenta amostras de 25 tipos de massas de treze marcas diferentes foram analisadas. A contagem de ECP de todas as amostras foi menor que 10 Unidades Formadoras de Colônia por grama (UFC/g, inferior a 5,0 × 10³ UFC/g, estabelecido como limite máximo pela legislação. As médias de a w das massas secas com e sem recheio variaram entre 0,70 e 0,61, respectivamente, o que impossibilita a multiplicação de ECP e produção de toxina. Por outro lado, a média de a w das massas frescas foi de 0,96, o que propicia o desenvolvimento desse padrão microbiológico. Os resultados obtidos neste trabalho indicam que a a w poderia ser utilizada como parâmetro para determinar em quais tipos de massas o padrão ECP deveria ser pesquisado, ou seja, para massas alimentícias cruas que possibilitassem a multiplicação de Staphylococcus spp. (a w > 0,86.
ANA P. BASSO
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.
Gartmeier, Martin; Lehtinen, Erno; Gruber, Hans; Heid, Helmut
Negative expertise is conceptualised as the professional's ability to avoid errors during practice due to certain cognitive agencies. In this study, negative knowledge (i.e. knowledge about what is wrong in a certain context and situation) is conceptualised as one such agency. This study compares and investigates the negative knowledge of elder…
Millar, Thomas J; Walsh, Catherine; Field, Thomas A
Until a decade ago, the only anion observed to play a prominent role in astrophysics was H - . The bound-free transitions in H - dominate the visible opacity in stars with photospheric temperatures less than 7000 K, including the Sun. The H - anion is also believed to have been critical to the formation of molecular hydrogen in the very early evolution of the Universe. Once H 2 formed, about 500 000 years after the Big Bang, the expanding gas was able to lose internal gravitational energy and collapse to form stellar objects and "protogalaxies", allowing the creation of heavier elements such as C, N, and O through nucleosynthesis. Although astronomers had considered some processes through which anions might form in interstellar clouds and circumstellar envelopes, including the important role that polycyclic aromatic hydrocarbons might play in this, it was the detection in 2006 of rotational line emission from C 6 H - that galvanized a systematic study of the abundance, distribution, and chemistry of anions in the interstellar medium. In 2007, the Cassini mission reported the unexpected detection of anions with mass-to-charge ratios of up to ∼10 000 in the upper atmosphere of Titan; this observation likewise instigated the study of fundamental chemical processes involving negative ions among planetary scientists. In this article, we review the observations of anions in interstellar clouds, circumstellar envelopes, Titan, and cometary comae. We then discuss a number of processes by which anions can be created and destroyed in these environments. The derivation of accurate rate coefficients for these processes is an essential input for the chemical kinetic modeling that is necessary to fully extract physics from the observational data. We discuss such models, along with their successes and failings, and finish with an outlook on the future.
Evaluation of a simplified key for the identification of coagulasepositive Staphylococcus isolated from bovine mastitis = Avaliação de uma chave de identificação simplificada para Staphylococcus coagulase-positivos isolados de mastite bovina
Ulisses de Pádua Pereira
Full Text Available Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis. Three hundred fourty four strains of coagulasepositive Staphylococcus (CPS, isolated from mastitis cases, underwent phenotypic and genotypictests to evaluate the efficiency of a simplified key, based on phenotypic tests for the discrimination of these microorganisms. The tests consisted of amplification of the femA gene and hemolysis in blood agar, production of acetoin and fermentation of maltose, mannitol and trehalose. Strains that showed negative results in the amplification test of the femA gene or that were not identified as Staphylococcus aureus (S. aureus by phenotypic tests were tested with the APISTAPH kit (Biomériux-France, for precise identification of species. Phenotypic tests revealed 338 strains (98.25% as S. aureus, three strains (0.86% as Staphylococcus hyicus, and threemicroorganisms (0.86% as Staphylococcus intermedius. PCR demonstrated that 338 (98.25% strains belonged to the S. aureus species, confirming the results for 336 strains from 338 identified, through a simplified phenotypic key. A high rate of correlation (98.83% was verifeid between the results of genotypic and phenotypic tests for the identification of S. aureus, demonstrating the applicabil