Assessment of antibacterial drug utilization patterns and antibiogram in infectious diseases: a prospective cross-sectional study.

BACKGROUND: Antibacterial drugs are successful in combating most types of infections. Irrational use and higher consumption of these drugs can give rise to the antibiotic resistance globally. OBJECTIVE: To evaluate antibacterial drug prescribing patterns and antibiogram in infectious disease cases admitted to the hospital. METHODS: A cross-sectional, observational study was conducted from September 2019 to February 2020 among inpatients ward at the hospital after ethical approval. All the data was analysed by the mean and percentage values using Microsoft excel. RESULTS: Out of 250 admitted patients, males and females were 156 (62%) and 94 (38%) respectively. The majority of patients 79 (32%) belonged to the age group of 20-40 years. The majority of prescriptions reported were for viral fever 48 (19%), lower respiratory tract infections 40 (16%) and dengue 33 (13%). Antibacterial drugs administered through the intravenous route and the oral route were 301 (83%) and 63 (17%) respectively. The most frequently utilized antibacterial drugs were beta-lactam class ceftriaxone 149 (60%) and the fixed-dose combination, amoxicillin plus clavulanic acid 65 (26%). Further highly prescribed antibacterial drugs were metronidazole 52 (21%), azithromycin 36 (15%), and levofloxacin 24 (10%). In Gram-negative bacteria, Escherichia coli 6 (30%) contributed majorly, while in Gram-positive coagulase-negative, Staphylococci 6 (30%) contributed the highest growth of bacteria for the specific infections in the admitted cases. CONCLUSION: Ceftriaxone and the amoxicillin-clavulanic acid combination were highly prescribed among all antibacterial drugs, followed by metronidazole and azithromycin. The current study showed that in the antibiogram pattern, Escherichia coli and coagulase-negative Staphylococci contributed significantly as causative organisms for infectious disease cases. The present study highlighted demographic distribution, infectious diseases with their antibacterial drug utilization patterns and antibiogram assessment in the admitted patients.

A real-world cost-effectiveness study of vancomycin versus linezolid for the treatment of late-onset neonatal sepsis in the NICU in China.

BACKGROUND AND OBJECTIVE: Currently, the detection rates of methicillin-resistant S. aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MRCoNS) in the blood cultures of neonates with sepsis exceed the national average drug resistance level, and vancomycin and linezolid are the primary antibacterial drugs used for these resistant bacteria according to the results of etiological examinations. However, a comprehensive evaluation of their costs and benefits in late-onset neonatal sepsis in a neonatal intensive care unit (NICU) has not been conducted. This study aimed to compare the cost and effectiveness of vancomycin and linezolid in treating neonatal sepsis in the NICU. METHODS: A cost-effectiveness analysis of real-world data was carried out by retrospective study in our hospital, and the cost and effectiveness of vancomycin and linezolid were compared by establishing a decision tree model. The drug doses in the model were 0.6 g for linezolid and 0.5 g for vancomycin. The cost break down included cost of medical ward, NICU stay, intravenous infusion of vancomycin or linezolid, all monitoring tests, culture tests and drugs. The unit costs were sourced from hospital information systems. The effectiveness rates were obtained by cumulative probability analysis. One-way sensitivity analysis was used to analyze uncertain influencing factors. RESULTS: The effectiveness rates of vancomycin and linezolid in treating neonatal sepsis in the NICU were 89.74% and 90.14%, respectively, with no significant difference. The average cost in the vancomycin group was ¥12261.43, and the average cost in the linezolid group was ¥17227.96. The incremental cost effectiveness was ¥12416.33 cost per additional neonate with treatment success in the linezolid group compared to vancomycin group at discharge. Factors that had the greatest influence on the sensitivity of the incremental cost-effectiveness ratio were the price of linezolid and the effectiveness rates. CONCLUSIONS: The cost for treatment success of one neonate in linezolid group was ¥5449.17 more than that in vancomycin group, indicating that vancomycin was more cost-effective. Therefore, these results can provide a reference for a cost effectiveness treatment scheme for neonatal sepsis in the NICU.

Microbiological monitoring of coagulase-negative Staphylococcus in public drinking water fountains: Pathogenicity factors, antimicrobial resistance and potential health risks.

The presence of opportunistic bacteria such as coagulase-negative Staphylococcus (CoNS) in drinking water poses public health concerns because of its potential to cause human infection and due to its antimicrobial resistance (AMR) diversity. This study evaluated the occurrence, virulence markers and AMR of CoNS in 468 drinking water samples from 15 public fountains located in four urban parks of São Paulo city (Brazil). Out of 104 samples positive for the presence of Staphylococcus genus, we detected CoNS in 75 of them (16%), which did not meet the Brazilian sanitary standards for residual chlorine. All isolates were of concern to public health for being responsible for infection in humans from low to high severity, nine of them are considered the most of concern due to 63.6% being multiresistant to antimicrobials. The results demonstrated that CoNS in drinking water must not be neglected. It is concluded that the presence of resistant staphylococci in drinking water is a potential health risk, which urges feasible and quick control measures to protect human health, especially in crowded public places.

Assessment of Coagulase-Negative Staphylococci and Lactic Acid Bacteria Isolated from Portuguese Dry Fermented Sausages as Potential Starters Based on Their Biogenic Amine Profile.

The aim of this study was to evaluate the decarboxylase activity of coagulase-negative staphylococci (CNS) and lactic acid bacteria (LAB) involved in meat products fermentation, in order to characterize and select the strains most suitable to be used as safe starter cultures. Isolates were obtained from traditional Portuguese dry fermented meat sausages, identified by PCR and characterized according to their technological properties. Lactobacilli and enterococci were assessed for their bacteriocinogenic potential. Biogenic amines (BA) were screened by culture method and analyzed by RP-HPLC/UV. The screening method, compared with chromatographic analysis, was not reliable for CNS and LAB strains selection. Tyramine decarboxylase activity was present in CNS strains, with a slight production of amines. No other hazardous BA were produced. Among lactobacilli, moderate production of tyramine was related only to Lactobacillus curvatus, with some strains producing putrescine or 2-phenylethylamine. Enterococci were high and moderate producers of tyramine and 2-phenylethylamine, respectively. Staphylococcus xylosus, Staphylococcus equorum, and Staphylococcus carnosus, independent of their genetic and technological profiles and BA production, were adequate for use in meat products, according to the data. Lactobacillus plantarum and Lactobacillus sakei strains could also be selected for starters. PRACTICAL APPLICATION: The selection of coagulase-negative staphylococci and lactic acid bacteria (LAB) isolates were based on their production of biogenic amines in order to avoid this potential hazard production in meat products. The most suitable isolates could be used as safe starter cultures in meat products industry. The staphylococci and LAB selected will achieve particular organoleptic characteristics in meat products and bioprotection from pathogens.

Assessment of the Dual Role of Clumping Factor A in S. Aureus Adhesion to Endothelium in Absence and Presence of Plasma.

Adhesion of Staphylococcus aureus to endothelial cells (ECs) is paramount in infective endocarditis. Bacterial proteins such as clumping factor A (ClfA) and fibronectin binding protein A (FnbpA) mediate adhesion to EC surface molecules and (sub)endothelial matrix proteins including fibrinogen (Fg), fibrin, fibronectin (Fn) and von Willebrand factor (vWF). We studied the influence of shear flow and plasma on the binding of ClfA and FnbpA (including its sub-domains A, A16+, ABC, CD) to coverslip-coated vWF, Fg/fibrin, Fn or confluent ECs, making use of Lactococcus lactis, expressing these adhesins heterologously. Global adherence profiles were similar in static and flow conditions. In the absence of plasma, L. lactis-clfA binding to Fg increased with shear forces, whereas binding to fibrin did not. The degree of adhesion of L. lactis-fnbpA to EC-bound Fn and of L. lactis-clfA to EC-bound Fg, furthermore, was similar to that of L. lactis-clfA to coated vWF domain A1, in the presence of vWF-binding protein (vWbp). Yet, in plasma, L. lactis-clfA adherence to activated EC-vWF/vWbp dropped over 10 minutes by 80% due to vWF-hydrolysis by a disintegrin and metalloproteinase with thrombospondin type 1 motif, member 13 and that of L. lactis-fnbpA likewise by > 70% compared to the adhesion in absence of plasma. In contrast, plasma Fg supported high L. lactis-clfA binding to resting and activated ECs. Or, in plasma S. aureus adhesion to active endothelium occurs mainly via two complementary pathways: a rapid but short-lived vWF/vWbp pathway and a stable integrin-coupled Fg-pathway. Hence, the pharmacological inhibition of ClfA-Fg interactions may constitute a valuable additive treatment in infective endocarditis.

Simple and economical method for identification and speciation of Staphylococcus epidermidis and other coagulase negative Staphylococci and its validation by molecular methods.

Coagulase-negative staphylococci (CoNS) have been increasingly recognized as a clinically important group of species that can cause several opportunistic nosocomial infections. There are at least 47 known species of Staphylococci and to differentiate all these species >40 biochemical tests need to be performed. The present study was able to refine the CoNS identification process by using only five tests to identify S. epidermidis from the rest and used six other tests to identify eleven other clinically significant CoNS species. A total of 242 CoNS isolates were collected from tertiary care hospitals and included in the study. The five-biochemical test scheme devised based on mathematical probability derived from a computer algorithm included fermentation of mannitol, maltose, mannose, trehalose and novobiocin susceptibility to differentiate S. epidermidis from other CoNS species. The remaining CoNS isolates other than S. epidermidis were further characterized with the help of six additional tests, which identified another eleven species. Species-specific PCR and 16SrDNA sequencing were used to confirm and validate the identification scheme. Species-specific PCR and 16SrDNA sequencing showed 100% agreement with non-divergent phenotypic test results, indicating that the five selected assays are highly specific for identifying S. epidermidis. In conclusion, this study used only 11 tests to identify most of the clinically significant CoNS that can reduce cost and time. This scheme is easy to perform in any laboratory with basic resources, the results of this study were validated using more accurate molecular methods such as PCR and 16S rDNA typing to confirm the utility of the proposed scheme.

La resistencia a los antibióticos y la falta de interés de la industria farmacéutica / Antibiotic resistance and the lack of interest by the pharmaceutical industry

La resistencia a los antibióticos aparece como consecuencia de mutaciones y de presiones de selección por el uso indiscriminado de estos medicamentos y se disemina rápidamente con alcance global. Esta diseminación se facilita por la poca higiene en los hospitales y por la mayor frecuencia en los viajes, el comercio y la trasmisión de las enfermedades. En Colombia, los primeros informes sobre resistencia aparecieron en 2001. El porcentaje de resistencia variaba según el microorganismo y el antibiótico. Por ejemplo, para ciprofloxacina en unidades de cuidados intensivos en Bogotá para Staphilococcus aureus (S. aureus) era del 60,0- 63,0%; para Staphilococcus coagulasa negativa (S. coagulasa negativa) del 47,0-58,0% y para Enterococcus sp. del 25,0- 44,0%. En hospitales de tercer nivel, se encontró resistencia del 52,0% para S. aureus y del 73,0% para S. aureus coagulasa negativa.

Resistance to antibiotics appears as a consequence of mutations and selection pressures due to the indiscriminate use of these drugs and spreads rapidly with global reach. This dissemination is facilitated by poor hygiene in hospitals and by the increased frequency of travel, trade and disease transmission. In Colombia, the first reports of resistance appeared in 2001. The percentage of resistance varied according to the microorganism and the antibiotic. For example, for ciprofloxacin in intensive care units in Bogota for Staphylococcus aureus (S. aureus) was 60.0- 63.0%; for coagulase-negative Staphylococcus (S. coagulase negative) 47.0-58.0% and for Enterococcus sp. 25.0- 44.0%. In tertiary hospitals, resistance was found to be 52.0% for S. aureus and 73.0% for S. aureus coagulase negative.

Assessment of the suitability of mannitol salt agar for growing bovine-associated coagulase-negative staphylococci and its use under field conditions.

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.

Assessment of the efficacy of polyclonal intravenous immunoglobulin G (IVIG) against the infectivity of clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) in vitro and in vivo.

The response to treatment of severe methicillin-resistant Staphylococcus aureus (MRSA) infections with the traditional antibiotics is sometimes unsatisfactory and multiple antibiotic resistance is common. Adjuvant therapy such as intravenous immunoglobulin G (IVIG) could possibly be helpful in the treatment of such infections. The effect of IVIG on the capacity of human neutrophils to phagocytose and kill MRSA was investigated in vitro using the MTT assay and measuring the production of reactive oxygen species (ROS) and nitric oxide (NO). The efficiency of IVIG in neutralizing α-hemolysin and coagulase of MRSA was also assessed. The capability of IVIG in the treatment and prevention of MRSA infections was also evaluated in a murine peritonitis model. IVIG significantly enhanced (p < 0.01) the killing of MRSA by neutrophils at all concentrations tested (0.1-5 mg/ml) by 30-80 % of control values. It significantly (p < 0.01) increased the level of NO production in a dose-dependent manner, giving up to 60 µM at 5 mg/ml. The ROS level significantly increased (p < 0.01) in the presence of IVIG. In addition, IVIG significantly reduced the hemolytic activity of MRSA 10-fold and its coagulation capabilities by 50 %. When tested in vivo, groups receiving IVIG via tail vein infusion showed no significant improvement in their survival. Only when delivered to the same site of infection did IVIG show an improvement in the survival of mice (n = 80). These results could pave the way for a better understanding of the mechanism of action of IVIG and suggest its clinical potential as an adjuvant preventive and therapeutic agent against life-threatening infections caused by MRSA and other bacteria.

Prevalence of antibiotic resistance in coagulase-negative staphylococci from spontaneously fermented meat products and safety assessment for new starters.

To provide new meat starter strains lacking antibiotic (AB) resistances, we explored the AB susceptibility in 116 coagulase-negative Staphylococcus (CNS) isolates from traditionally fermented sausages (n=40) manufactured with meat from conventional animal breeding, and from meat products (n=76) made from meat of animals raised in natural habitats under low- or no-antibiotic pressure. Less than 50% of these CNS isolates showed phenotypic resistances to at least one antibiotic (AB) by using microdilution assay. Resistances to penicillins and tetracycline were most often observed and could be traced back to blaZ and tet(K) genes. Prevalence of AB resistances was species-dependent and mainly found in isolates of Staphylococcus warneri (78%), Staphylococcus capitis (75%) and Staphylococcus epidermidis (67%), but only sporadically detected in Staphylococcus carnosus (27%) and Staphylococcus equorum (18%). AB resistances were more often observed in S. xylosus isolates originating from natural habitats compared to traditionally fermented sausages made from conventional meat. A selection of 101 isolates belonging to S. xylosus (n=63), S. carnosus (n=21) and S. equorum (n=17) were subsequently grouped by pulsed-field gel electrophoresis (PFGE) into strain clusters. No S. carnosus and only five S. xylosus strains were lacking AB resistances and exhibited a PFGE genotype different from commercial starters. These strains, together with 17 S. equorum strains, were further studied for safety and technological characteristics. The ability to produce biogenic amines was not detected in any strain. PCR amplifications for enterotoxin encoding genes seg-sej were detected in one, and for δ-hemolysin encoding gene hld in four S. equorum strains, but phenotypic hemolytic activity was visible for three S. xylosus and 15 S. equorum strains. Catalase and nitrate reductase activity was observed in all isolates tested; particularly S. equorum showed high nitrate reduction. In conclusion, we were able to select four new meat starter strains (two S. xylosus and two S. equorum strains) out of 116 investigated CNS, fulfilling all safety criteria including the absence of AB resistances, production of biogenic amines and genes encoding virulence factors but exhibiting high nitrate reductase and catalase activity as suitable technological characteristics. Thus, S. equorum isolates, often the dominant species in spontaneously fermented meat products, provided a prospective meat starter species exhibiting high nitrate reduction and low prevalence of AB resistances.

Comparison of bacterial identification by MALDI-TOF mass spectrometry and conventional diagnostic microbiology methods: agreement, speed and cost implications.

Identification of microbial pathogens still relies primarily on culture and phenotypic methods, which is labour-intensive and time-consuming. In this study, identification of bacteria with valid standard identification using BD Phoenix, API panels and other recommended procedures is compared to identification with matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF) mass spectrometry using the MALDI Biotyper (Bruker Daltonics) in the setting of a routine NHS diagnostic microbiology laboratory. In total, 928 bacterial isolates obtained from blood (n=463), wounds and pus (n=208), respiratory tract (n=100), faeces (n=86) and urines (n=71) were analysed. There were 721 (77.7%) isolates with a MALDI Biotyper score > or =2.0, indicating secure genus and probable species identification; and 149 (16.1%) isolates with a score > or =1.7 and <2.0 indicating probable genus identification. The isolates with scores of > or =2.0 and > or =1.7 comprised 31 and 33 genera and 65 and 67 species, respectively. Overall, 99.4% and 99.1% of organism identifications were in agreement between the MALDI Biotyper and conventional identification at the genus level, and 89.3% and 87.8% at species level when analysing organisms with MALDI Biotyper scores > or =2.0 and > or =1.7, respectively. With many but not all organisms, identification at the genus level is sufficient; however, MALDI Biotyper separation of 208 staphylococci into Staphylococcus aureus and coagulase-negative staphylococci was always correct when scores were > or =1.7. First results were obtained after 5-10 min and analysis of a full 96-well target plate was completed in approximately 90 min. Substantial savings of between pounds 1.79 and pounds 2.56 per isolate, depending on the cost model of acquisition of the MALDI Biotyper system and number of isolates tested, would be realised when all 928 isolates were identified using the MALDI Biotyper and disk-susceptibility testing when compared to the cost for 618 Phoenix ID panels and 158 API panels and disk-susceptibility tests only (i.e., not taking into account costs incurred for identification of the remaining 152 mixed isolates). Microbial identification by MALDI Biotyper offers a rare opportunity for significant cost-neutral or even cost-saving quality improvements in medical diagnostics.

Assessment of impact of peptide nucleic acid fluorescence in situ hybridization for rapid identification of coagulase-negative staphylococci in the absence of antimicrobial stewardship intervention.

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.

Pharmacoeconomic analysis of microbiologic techniques for differentiating staphylococci directly from blood culture bottles.

Differentiating staphylococci in blood cultures is a critical issue, particularly when only one of two cultures is positive by Gram staining for staphylococci. New tests for the identification of Staphylococcus aureus allow faster results and definitive treatment compared to the tube coagulase test interpreted at 24 h (TCT24). These newer tests, peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) and real-time PCR (RT-PCR), offer improved sensitivity at higher cost. Data suggest that the tube coagulase test may be interpreted at 4 h (TCT4) with little loss of sensitivity. The impact of variability in turnaround time, sensitivity, specificity, and cost on comparative cost-effectiveness is unknown. Our aim was to establish the cost-effectiveness of TCT24, PNA-FISH, RT-PCR, and TCT4 for direct identification of staphylococci in blood cultures. Decision analysis comparing these strategies was done from the institutional perspective. Besides test variables, other variables included patient risk factors, empirical treatment, and follow-up cultures. Probability and cost estimates came from the literature and institutional data. Base case estimates were derived from institutional rates of 73% contamination when coagulase-negative staphylococci were identified, 67.6% prevalence of risk factors, and 12.4% prevalence of S. aureus when one of two cultures yielded staphylococci. Sensitivity analysis was done across a range of probabilities and costs. In the base case, TCT4 and TCT24 were more cost-effective than RT-PCR and PNA-FISH ($78 versus $120 versus $165 per patient, respectively). The advantage of TCT4 and TCT24 remained robust upon sensitivity analysis. TCT4 should be further evaluated as a rapid, cost-effective means for identification of S. aureus in blood cultures.

Pharmacodynamic comparison of linezolid, teicoplanin and vancomycin against clinical isolates of Staphylococcus aureus and coagulase-negative staphylococci collected from hospitals in Brazil.

Pharmacodynamic exposures, measured as the ratio of steady-state total drug area under the curve to MIC (AUC/MIC), were modelled using a 5000-patient Monte-Carlo simulation against 119 non-duplicate clinical isolates of Staphylococcus aureus and 82 coagulase-negative staphylococci (CNS) collected from hospitals in Brazil between 2003 and 2005. Pharmacodynamic targets included an AUC/MIC >82.9 for linezolid and >345 for teicoplanin and vancomycin, as well as a free drug AUC/MIC >180 for vancomycin. The cumulative fractions of response (CFRs) against all S. aureus isolates were 96.0%, 30.1%, 71.6%, 48.0% and 65.1% for linezolid 600 mg every 12 h, teicoplanin 400 mg every 24 h and 800 mg every 24 h, and vancomycin 1000 mg every 12 h and every 8 h, respectively. Using a free drug target for vancomycin improved the CFR to 94.6% for the high-dose regimen, but did not substantially alter results for the lower dose. CFRs against all CNS isolates were 97.8%, 13.4%, 34.6%, 10.9% and 31.3%, respectively, for the same antibiotic regimens. The CFR was reduced for all compounds among the methicillin-resistant isolates, except for linezolid against methicillin-resistant CNS. Sensitivity analyses did not alter the final order of pharmacodynamic potency against these isolates. Although higher doses of vancomycin and teicoplanin increased the CFR, the likelihood of achieving bactericidal targets was still lower than with linezolid. The results for the high-dose vancomycin regimen were highly dependent on the pharmacodynamic target utilised. These data suggest that linezolid has a greater probability of attaining its requisite pharmacodynamic target than teicoplanin and vancomycin against these staphylococci.

Safety assessment of dairy microorganisms: coagulase-negative staphylococci.

The genus Staphylococcus is made up of 36 validated species which contain strains that are pathogenic, saprophytic, or used as starter cultures for the food industry. Staphylococci species used in cheese-making are novobiocin-resistant, coagulase-negative and are not usually identified at species level by routine laboratories. A bibliographic survey was conducted to assess safety status of CNS used in fermented dairy foods. Commercial kits based on phenotypic discrimination can't reliably identify these species because of the variable expression of some phenotypic traits. Several molecular targets, including 16S rRNA, hsp60, tuf, SodA and rpoB genes can be used for identifying species of the Staphylococcus genus. No coagulase-negative staphylococci isolated from milk or dairy products has ever been involved in a case of food poisoning or human pathology following ingestion of dairy products. Nevertheless, a few case of nosocomial infection caused by some species (S. caprae, S. capitis, S. sciuri) have been reported in patients with depressed immune systems or who have undergone long severe hospital treatments, or in the presence of an indwelling catheter or foreign materials such as cardiac prostheses. They may therefore be regarded as exceptional opportunistic pathogens in certain clinical situations.

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

OBJECTIVES: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Detection of slime production by coagulase-negative Staphylococcus: an assessment of two methods.

The detection of slime can be helpful in determining the diagnostic value of coagulase-negative Staphylococcus (CNS). Here, we have made a comparative study of two phenotypic methods of slime production test from 28 clinically significant blood culture isolates of CNS. A total 16 out of 28 isolates (57.1%) were positive for slime by either of the tests, whereas only 5 out of 28 isolates (17.9%) were positive for slime by both the tests. The tube method detected slime in significantly more number of isolates than the spectrophotometric method (14 vs. 7; p<0.0483279). A weak correlation was found between the tube method and the spectrophotometric method (rs=0.2474). The tube test was found to be superior to the spectrophotometric test in terms of sensitivity, ease of performance and interpretation, and cost effectiveness.

Predictive value of blood cultures positive for coagulase-negative staphylococci: implications for patient care and health care quality assurance.

Interpretation of blood cultures that are positive for coagulase-negative staphylococci (CoNS) is often difficult. Predictive values for blood cultures positive for CoNS in patients with a central vascular line in place were calculated using the following rates: true bacteremia, 3%; blood culture contamination, 2%; detection of bacteremia, 80%; and catheter colonization, 2% (for blood samples obtained through a central vascular line). Positive predictive values were 55% for 1 positive culture result of 1 culture performed, 20% for 1 positive result of 2 performed, and only 5% for 1 positive result of 3 performed. For 2 positive culture results of 2 cultures performed, the positive predictive value was 98% if both samples were obtained through the vein, 96% if one sample was obtained through a catheter and the other was obtained by vein, and only 50% if both samples were obtained through a catheter. Use of this model with institution-specific values for input parameters would assist in clinical decision-making as well as hospital quality assurance.

Evaluation of three rapid methods for the direct identification of Staphylococcus aureus from positive blood cultures.

Staphylococci represent the most commonly encountered blood culture isolates. Differentiating Staphylococcus aureus from coagulase-negative staphylococci (CoNS) is important in guiding empirical therapy, especially since the majority of CoNS are contaminants. This study evaluated three rapid methods for the direct identification of S. aureus from blood cultures. A total of 157 patient blood cultures with gram stains showing gram-positive cocci in clusters were included. The following assays were evaluated: API RAPIDEC staph (API) (bioMerieux, Durham, N.C.), the tube coagulase test (TCT) read at 4 h, and peptide nucleic acid (PNA) fluorescence in situ hybridization (FISH) (AdvanDx, Woburn, Mass.). All assays yielded results of S. aureus or non-S. aureus. The direct rapid results were compared to results obtained with isolated colonies using the AccuProbe Staphylococcus aureus Culture Identification Test (Gen-Probe, San Diego, Calif.). API, TCT, and PNA FISH exhibited sensitivities of 96, 84, and 99% and specificities of 99, 100, and 100%, respectively. Direct identification testing by any of these three assays yielded acceptable performance and timely results. This ability to accurately detect S. aureus in blood cultures gives the physician information with which to initiate or tailor antimicrobial therapy. Coupled with direct susceptibility testing of positive blood culture broths, the patient and institution may experience improved outcomes.

Assessment of the aerobic faecal microflora in mink (Mustela vison Schreiber) with emphasis on Escherichia coli and Staphylococcus intermedius.

The present study was undertaken to investigate the culturable aerobic faecal microflora of mink from newborn until adulthood with emphasis on the potential pathogens Escherichia coli and beta-haemolytic coagulase positive staphylococci. Rectal swabs were taken from 10 healthy dams and their offspring on seven mink farms throughout the production season and a semi-quantitative enumeration of total E. coli and haemolytic E. coli, beta-haemolytic streptococci, beta-haemolytic coagulase positive staphylococci, total lactic acid bacteria, and enterococci was carried out in all samples using selective and non-selective media. Aerobic bacteria were cultured from close to 100% of the samples throughout the survey. Prevalence of E. coli isolates varied between 70 and 90% of the samples throughout the survey with a small decline at the end of the study period. The highest bacterial counts were found among recently weaned kits or kits in the early growth period (P<0.0012). Lactic acid bacteria and enterococci were isolated from more than 90% of all samples, while beta-haemolytic staphylococci were isolated from 20 to 70% of the samples. While beta-haemolytic staphylococci were dominant from birth and during the nursing period, counts of staphylococci gradually decreased during the nursing period and were outnumbered by E. coli during the growth season.