In-Vitro Assessment Of The Therapeutic Potential Of Polymyxins And Tigecycline Against Multidrugresistant Acinetobacter Isolates From Infected Wounds.

BACKGROUND: The incidence of multidrug-resistant (MDR), extreme drug resistant (XDR), and pan drug-resistant (PDR) Acinetobacter are increasing throughout the world. The therapeutic management and control of Acinetobacter are difficult due to the emergence of drug resistance and its enduring capacity to survive in the environment. The present study was designed to appraise the efficacy of Polymyxins and Tigecycline against multidrugresistant Acinetobacter isolates from surgical and burn wounds. METHODS: During the study, the specimens were collected from various types of wounds from inpatients and outpatients of the tertiary care hospitals of Lahore, Pakistan in 2017 and 2018. The bacterial pathogens were isolated and identified using standard microbiological procedures and molecular confirmation of Acinetobacter species was examined by PCR using specific primers. The antibiotic susceptibility profiling of Acinetobacter isolates was studied against 18 antibiotics as per Clinical and Laboratory Standards Institute (CLSI) guidelines. RESULTS: The Acinetobacter isolates demonstrated extreme resistance especially to ampicillin/sulbactam, piperacillin/tazobactam, cephalosporins, carbapenems, fluoroquinolones, and aminoglycosides. However, the colistin, polymyxin, and tigecycline remained the most effective antimicrobial agents against Acinetobacter isolates. CONCLUSIONS: The results highlight the extent of drug resistance and therapeutic potential of Polymyxins and Tigecycline for wound infections caused by MDR and XDR Acinetobacter species. The wiser use of antimicrobials, incessant surveillance of antimicrobial resistance, and stringent adherence to infection control guidelines are critical to reducing major outbreaks in the future.

Individual and combined mechanistic toxicity of sulfonamides and their implications for ecological risk assessment in the Three Gorges Reservoir Area (TGRA), China.

Sulfonamides (SAs) are conventional veterinary antibiotics that pose ecological risks in the aquatic environment. This study aims to evaluate the environmental concerns of SAs in the Three Gorges Reservoir Area (TGRA) and their toxicogenetic implications. Here, we employed various in vitro and in vivo bioassays to determine the combine toxicogenetic effects of SAs, which were further confirmed through applying Combination Index (CI) and Independent Action (IA) models. Among the investigated SAs, sulfamethoxazole (SMX) appeared as the individual chemical with relatively high environmental effects and elevated in vitro and in vivo toxicity. Importantly, exposure to the binary mixtures of SAs induced higher developmental toxicity and significantly perturbed the detoxification pathway in zebrafish, compared to that of individual compound exposure. Moreover, the CI and IA models indicated greater synergistic effects of SAs binary mixtures as SMX-SMR, SMX-ST, and SPY-ST on the Acinetobacter sp. Tox2 at Fa = 0.5. Contrarily, IA model predicted the additive, antagonistic and synergistic effects of SAs mixtures on the transcriptional responses of detoxification pathways in zebrafish, implying the different mode of actions (MoAs) for SAs to induce mixture toxicity in vivo. Thus, the nature of toxicological interactions of SAs should be considered while performing their ecological risk assessment.

Acinetobacter spp in a Third World Country with Socio-economic and Immigrants Challenges.

INTRODUCTION: In the last decade, Acinetobacter species have taken a major public health concern. This is mainly due the increased resistance to a wide range of antibiotics causing treatment challenges. In view of the constant population mobilization and the economic crisis that Lebanon is currently facing, it becomes a necessity to re-evaluate the real threat of Acinetobacter spp and its implication in the one health. METHODOLOGY: This review was conducted through the analysis of 45 research papers and reports pertaining to Acinetobacter spp performed in Lebanon. More than 82% of the papers consulted were published in international journals and more than 70 percent of them had received impact factor. RESULTS: An in depth description of the involvement of this organism in human infection and its role as potential pathogen or simple colonizer was performed. In addition, the different aspects of resistance, mostly to carbapenems and colistin was studied and summarized. While in animals and environment, susceptible strains were mostly isolated, OXA-23/OXA-24 were predominant in humans. Recently, NDM-1 producing Acinetobacter spp was detected in a Syrian refugee which then was reported in Lebanese patients. The bacterial identification procedures are non-systematic and not always reliable in the Lebanese studies presenting sometimes discrepancies an inconsistency. CONCLUSION: Acinetobacter is commonly isolated Lebanon. In view of the spread of resistance among these isolated and their dissemination, Infection control measures attempting to control the spread of this genus in and outside hospitals are lacking and thus require more attention and stewardship activities.

Improvement of gram-negative susceptibility to fluoroquinolones after implementation of a pre-authorization policy for fluoroquinolone use: A decade-long experience.

OBJECTIVE: Due to concerns over increasing fluoroquinolone (FQ) resistance among gram-negative organisms, our stewardship program implemented a preauthorization use policy. The goal of this study was to assess the relationship between hospital FQ use and antibiotic resistance. DESIGN: Retrospective cohort. SETTING: Large academic medical center. METHODS: We performed a retrospective analysis of FQ susceptibility of hospital isolates for 5 common gram-negative bacteria: Acinetobacter spp., Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Primary endpoint was the change of FQ susceptibility. A Poisson regression model was used to calculate the rate of change between the preintervention period (1998-2005) and the postimplementation period (2006-2016). RESULTS: Large rates of decline of FQ susceptibility began in 1998, particularly among P. aeruginosa, Acinetobacter spp., and E. cloacae. Our FQ restriction policy improved FQ use from 173 days of therapy (DOT) per 1,000 patient days to <60 DOT per 1,000 patient days. Fluoroquinolone susceptibility increased for Acinetobacter spp. (rate ratio [RR], 1.038; 95% confidence interval [CI], 1.005-1.072), E. cloacae (RR, 1.028; 95% CI, 1.013-1.044), and P. aeruginosa (RR, 1.013; 95% CI, 1.006-1.020). No significant change in susceptibility was detected for K. pneumoniae (RR, 1.002; 95% CI, 0.996-1.008), and the susceptibility for E. coli continued to decline, although the decline was not as steep (RR, 0.981; 95% CI, 0.975-0.987). CONCLUSIONS: A stewardship-driven FQ restriction program stopped overall declining FQ susceptibility rates for all species except E. coli. For 3 species (ie, Acinetobacter spp, E. cloacae, and P. aeruginosa), susceptibility rates improved after implementation, and this improvement has been sustained over a 10-year period.

Trends and factors associated with antimicrobial resistance of Acinetobacter spp. invasive isolates in Europe: A country-level analysis.

OBJECTIVES: This study investigated trends and factors associated with antimicrobial resistance (AMR) in Acinetobacter spp. in Europe. METHODS: Using data from EARS-Net, population-weighted multilevel logistic regression models with random intercepts for each participating country were performed to assess trends in Acinetobacter AMR. Countries were divided into two groups (Northern versus Southern-Eastern) using a convenient US$35000 cut-off of the 2016 gross domestic product per capita (GDPPC). RESULTS: In most countries, there were no ascending or descending trends over time. The models showed a consistent higher prevalence of AMR to aminoglycosides, carbapenems and fluoroquinolones in countries with GDPPC US$35000 and

A novel GoldNano Carb test for rapid phenotypic detection of carbapenemases, particularly OXA type, in Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter spp.

Objectives: To develop a simple gold nanoparticle (AuNP)-based colorimetric test, GoldNano Carb (GoldC), for detecting carbapenemase production in Gram-negative bacteria, compared with updated Carba NP (CNP) and CarbAcineto NP (CAcNP) tests by using PCR methods as gold standard. Methods: Ninety-nine carbapenemase-producing Enterobacteriaceae (CPE), Pseudomonas spp. and Acinetobacter spp. isolates and 89 non-CPE isolates were tested by the GoldC and CNP. Additionally, the CAcNP was performed in the Acinetobacter spp. isolates. The final imipenem (imipenem/cilastatin form) concentration was 5 mg/mL for all three tests. For the GoldC, the imipenem powder was added directly to bacterial cell suspension in distilled water prior to detection of acid product by the citrate-capped AuNP solution. An AuNP change from red to purple, blue or green indicates carbapenemase activity. Results: The GoldC detected all carbapenemase producers except one OXA-23-like producer (99.0% sensitivity), whereas 11 carbapenemase producers (10 Acinetobacter and 1 P. aeruginosa) were CNP negative (88.9% sensitivity). However, the GoldC and CNP provided 100% and 98.6% sensitivity, respectively, for the CPE and Pseudomonas spp. Both tests gave one false positive from CTX-M-1-like-producing Enterobacter spp. (98.9% specificity). The GoldC and CAcNP detected 96.7% and 93.3% of the Acinetobacter spp. isolates, respectively. Interestingly, times to positivity by the GoldC were markedly shorter than those by the CNP (76.8% versus 36.2% positive at 5 min) and CAcNP (43.3% at 5 min versus 20% within 30 min). Conclusions: The GoldC is fast, easy, highly sensitive and inexpensive (∼$0.25 per test), suggesting that it may be suitable for routine carbapenemase detection in low-resource settings for infection control or epidemiological purposes.

Costs and Mortality Associated With Multidrug-Resistant Healthcare-Associated Acinetobacter Infections.

BACKGROUND Our objective was to estimate the per-infection and cumulative mortality and cost burden of multidrug-resistant (MDR) Acinetobacter healthcare-associated infections (HAIs) in the United States using data from published studies. METHODS We identified studies that estimated the excess cost, length of stay (LOS), or mortality attributable to MDR Acinetobacter HAIs. We generated estimates of the cost per HAI using 3 methods: (1) overall cost estimates, (2) multiplying LOS estimates by a cost per inpatient-day ($4,350) from the payer perspective, and (3) multiplying LOS estimates by a cost per inpatient-day from the hospital ($2,030) perspective. We deflated our estimates for time-dependent bias using an adjustment factor derived from studies that estimated attributable LOS using both time-fixed methods and either multistate models (70.4% decrease) or matching patients with and without HAIs using the timing of infection (47.4% decrease). Finally, we used the incidence rate of MDR Acinetobacter HAIs to generate cumulative incidence, cost, and mortality associated with these infections. RESULTS Our estimates of the cost per infection were $129,917 (method 1), $72,025 (method 2), and $33,510 (method 3). The pooled relative risk of mortality was 4.51 (95% CI, 1.10-32.65), which yielded a mortality rate of 10.6% (95% CI, 2.5%-29.4%). With an incidence rate of 0.141 (95% CI, 0.136-0.161) per 1,000 patient-days at risk, we estimated an annual cumulative incidence of 12,524 (95% CI, 11,509-13,625) in the United States. CONCLUSION The estimates presented here are relevant to understanding the expenditures and lives that could be saved by preventing MDR Acinetobacter HAIs. Infect Control Hosp Epidemiol 2016;1-7.

Initial Assessment of the Molecular Epidemiology of blaNDM-1 in Colombia.

We report complete genome sequences of four blaNDM-1-harboring Gram-negative multidrug-resistant (MDR) isolates from Colombia. The blaNDM-1 genes were located on 193-kb Inc FIA, 178-kb Inc A/C2, and 47-kb (unknown Inc type) plasmids. Multilocus sequence typing (MLST) revealed that these isolates belong to sequence type 10 (ST10) (Escherichia coli), ST392 (Klebsiella pneumoniae), and ST322 and ST464 (Acinetobacter baumannii and Acinetobacter nosocomialis, respectively). Our analysis identified that the Inc A/C2 plasmid in E. coli contained a novel complex transposon (Tn125 and Tn5393 with three copies of blaNDM-1) and a recombination "hot spot" for the acquisition of new resistance determinants.

The Cost of Responding to an Acinetobacter Outbreak in Critically Ill Surgical Patients.

BACKGROUND: Our institution had an outbreak of multi-drug-resistant Acinetobacter (MDRA) in 2011. We analyzed the costs of responding to this outbreak from the hospital's perspective. METHODS: We estimated retrospectively the excess costs associated with an MDRA outbreak response at a major academic medical center, including the costs of staffing, supplies, administrative time, deep cleaning, and environmental testing. Differences in mean costs before and during the 2011 MDRA outbreak were analyzed using the Student t-test. RESULTS: The overall excess cost incurred during the outbreak response was $371,079 in 2011 U.S. dollars. The largest contributors were the extra resources needed to staff and clean the two intensive care units (ICUs) (78%). In the general surgery ICU, the mean weekly cost of nursing during the outbreak was $13,276 more for regular hours (+15%; p < 0.01) than in the pre-outbreak period and $2,682 more for overtime hours (+86%; p = 0.02). In the trauma ICU, the cost was $20,746 more for regular hours (+24%; p < 0.01) and $3,445 more for overtime hours (+124%; p < 0.01). The costs of supplies ($13,036; +30%; p = 0.03) and gloves ($2,572; +48%; p = 0.01) also were greater during the outbreak. Administrative time, consumables, use of a surge pod, and environmental testing accounted for the remainder of the extra costs. CONCLUSIONS: Our institution incurred $371,079 in excess costs as a result of an MDRA outbreak. This figure does not include the costs related to treatment of the infections, loss of reimbursement because of hospital-acquired infection, legal services, or changes in staff morale, patient satisfaction, or hospital reputation. Strategies to prevent and control such outbreaks better have substantial value.

Population pharmacokinetics and dosing simulations of imipenem in serious bacteraemia in immunocompromised patients with febrile neutropenia.

The aims of this study were to i) reveal the population pharmacokinetics; and ii) assess the probability of target attainment (PTA) and cumulative fraction of response (CFR) (defined as the expected population PTA for a specific drug dose and a specific population of microorganisms) of imipenem in febrile neutropenic patients with bacteraemia. Ten patients were randomised into two groups: Group I received a 0.5-h infusion of 0.5 g of imipenem every 6 h (q6h) for 8 doses; and Group II received a 4-h infusion of 0.5 g q6h for 8 doses. A Monte Carlo simulation was performed to determine the PTA. The volume of distribution and total clearance of imipenem were 20.78 ± 1.35 l and 23.19 ± 1.34 l/h, respectively. Only a 4-h infusion of 1 g q6h regimen achieved a PTA >93% for 80% T>MIC for a MIC of 2 µg/ml. A 4-h infusion of all simulated regimens and a 0.5-h infusion of 0.5 g q6h and 1 g q6h achieved targets (CFR ≥ 90%) against Escherichia coli and Klebsiella spp. However, against Pseudomonas aeruginosa and Acinetobacter spp., no regimens achieved their targets. In conclusion, the results indicate that a higher than manufacturer's dosage recommendation is required to maximize the activity of imipenem.

Toxicity assessment and modelling of Moringa oleifera seeds in water purification by whole cell bioreporter.

Moringa oleifera has been used as a coagulation reagent for drinking water purification, especially in developing countries such as Malawi. This research revealed the cytoxicity and genotoxicity of M. oleifera by Acinetobacter bioreporter. The results indicated that significant cytoxicity effects were observed when the powdered M. oleifera seeds concentration is from 1 to 50 mg/L. Through direct contact, ethanolic-water extraction and hexane extraction, the toxic effects of hydrophobic and hydrophilic components in M. oleifera seeds were distinguished. It suggested that the hydrophobic lipids contributed to the dominant cytoxicity, consequently resulting in the dominant genotoxicity in the water-soluble fraction due to limited dissolution when the M. oleifera seeds granule concentration was from 10 to 1000 mg/L. Based on cytoxicity and genotoxicity model, the LC50 and LC90 of M. oleifera seeds were 8.5 mg/L and 300 mg/L respectively and their genotoxicity was equivalent to 8.3 mg mitomycin C per 1.0 g dry M. oleifera seed. The toxicity of M. oleifera has also remarkable synergistic effects, suggesting whole cell bioreporter as an appropriate and complementary tool to chemical analysis for environmental toxicity assessment.

The impact of a nationwide antibiotic restriction program on antibiotic usage and resistance against nosocomial pathogens in Turkey.

PURPOSE: Antimicrobial resistance among microorganisms is a global concern. In 2003, a nationwide antibiotic restriction program (NARP) was released in Turkey. In this study we evaluated the effect of NARP on antibiotic consumption, antimicrobial resistance, and cost. MATERIALS AND METHODS: The data obtained from all of the four university hospitals, and one referral tertiary-care educational state hospital in Ankara. Antimicrobial resistance profiles of 14,233 selected microorganisms all grown in blood cultures and antibiotic consumption from 2001 to 2005 were analyzed retrospectively. RESULTS: A negative correlation was observed between the ceftriaxone consumption and the prevalence of ceftriaxone resistant E.coli and Klebsiella spp. (rho:-0.395, p:0.332 and rho:-0.627, p:0.037, respectively). The decreased usage of carbapenems was correlated with decreased carbapenems-resistant Pseudomonas spp. and Acinetobacter spp (rho:0.155, p:0.712 and rho:0.180, p:0.668, respectively for imipenem). Methicillin resistance rates of S.aureus were decreased from 44% to 41%. After two years of NARP 5,389,155.82 USD saving occurred. CONCLUSION: NARP is effective in lowering the costs and antibiotic resistance.

Comprehensive assessment of tigecycline activity tested against a worldwide collection of Acinetobacter spp. (2005-2009).

A total of 5127 Acinetobacter spp. were collected from 140 hospitals in 32 countries in North America (17.1%), Europe (22.9%), Latin America (25.2%), and the Asia-Pacific (APAC) region (34.8%). Tigecycline MIC distributions were bimodal against isolates from North America and APAC region, while a unimodal pattern was noted for strains from Latin America. A variable MIC distribution was noted in Europe. Only tigecycline (MIC(50/90), 0.5/2 µg/mL) and polymyxin B (MIC(50/90), 0.5/1 µg/mL; 98.6% susceptible) exhibited high activity against Acinetobacter spp. Overall, tigecycline inhibited at least 90.0% of Acinetobacter spp. isolates from all countries evaluated at ≤2 µg/mL, as well as 95.0% of those displaying multidrug resistance. Other tested agents showed limited activity and a significant (P < 0.001) trend toward decreased susceptibility during the study period.

Evaluating empiric treatment options for secondary peritonitis using pharmacodynamic profiling.

BACKGROUND: Selecting an appropriate agent for empiric antibiotic therapy for secondary peritonitis is challenging. The pathogens responsible, aerobic gram-negative bacilli in particular, are becoming more resistant to antibiotics. The purpose of this study was to predict the ability of common antimicrobial regimens to achieve optimal pharmacodynamic exposure against aerobic bacteria implicated in secondary peritonitis, while considering current national resistance trends. METHODS: Monte Carlo simulation was used to model pharmacodynamic endpoints and compare the cumulative fraction of response (CFR) for imipenem-cilastatin, meropenem, ertapenem, piperacillin/tazobactam, ceftazidime, ceftriaxone, ciprofloxacin, and levofloxacin against isolates of species associated with secondary peritonitis. Minimum inhibitory concentration (MIC) distributions for isolates collected in North America were obtained from the 2004 MYSTIC database. Pharmacokinetic parameters were derived from the literature; the endpoints evaluated included free drug time above the MIC (fT(>MIC)) and the area under the concentration-time curve to MIC ratio (AUC:MIC). RESULTS: The simulation predicted that several compounds would have a superior probability of providing appropriate coverage of aerobic bacteria: Imipenem-cilastatin (98.6% CFR at 1 g q8h), meropenem (98.2% CFR at 1 g q8h), ertapenem (91.7% CFR at 1 g q24h), piperacillin/ tazobactam (93.7% CFR at 3.375 g q6h), ceftazidime (91.1% CFR at 2 g q8h), and cefepime (92.9% CFR at 1 g q12h and 95.8% CFR at 2 g q12h). Ceftriaxone, ciprofloxacin, and levofloxacin exhibited CFRs < 82%. CONCLUSIONS: Considering contemporary susceptibility data for aerobic bacteria, monotherapy with any of the three carbapenems or piperacillin/tazobactam 3.375 g q6h would provide optimal exposure for the pathogens commonly encountered in secondary peritonitis. Cefepime (in combination with metronidazole to provide anti-anaerobic coverage) also would be an acceptable choice, as would ceftazidime given at 2 g q8h (again in combination with metronidazole). Despite the popularity of combination therapy based on ciprofloxacin, levofloxacin, or ceftriaxone with metronidazole, these choices appear to be inferior to the other options because of emerging antibiotic resistance, particularly in E. coli.

Assessment of biofilm cell removal and killing and biocide efficacy using the microtiter plate test.

Biofilm formation on surfaces has serious economic and environmental implications. Growth of biofilm within a water distribution system can lead to problems such as biocorrosion and biofouling accumulation. To prevent and control these occurrences, it is necessary to use suitable biocides to remove the biofilm and kill biofilm cells. In this study, the genera Actinobacillus, Branhamella, Bacillus, Micrococcus and Acinetobacter were isolated from biofilms formed on brass coupons exposed to a cooling water system. It was shown by the microtiter plate test that a mixed culture of the isolates and a single culture of Acinetobacter sp(2) produced high levels of biofilm formation. A microwell plate technique was applied for assessment of the ability of various biocides to remove and kill mixed-culture biofilm cells and Acinetobacter sp(2), the latter as a single-species biofilm with a high rate of biofilm production. The results showed that the mixed-culture biofilm cells had more resistance to removal and killing by some biocides, such as hydrogen peroxide and sulfathiazole, than the single-species biofilm cells (Acinetobacter sp(2)). Oxidising biocides, such as sodium hypochlorite and hydrogen peroxide, demonstrated a higher potential for biofilm removal and killing compared with non-oxidising biocides (sulfathiazole and glutaraldehyde).

The cost of antibiotic resistance: effect of resistance among Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudmonas aeruginosa on length of hospital stay.

To assess the effect of antimicrobial resistance on length of hospital stay, a case-control study compared infections due to four nosocomial pathogens. Significantly increased lengths of stay were associated with infections due to methicillin-resistant Staphylococcus aureus, extended-spectrum beta-lactamase-producing Klebsiella pneumoniae, and carbapenem-resistant Acinetobacter baumannii or Pseudomonas aeruginosa. Infections with resistant pathogens are associated with prolonged hospitalization.

Antibiotic resistance in Acinetobacter spp. isolated from sewers receiving waste effluent from a hospital and a pharmaceutical plant.

The possible increase of antibiotic-resistant bacteria in sewage associated with the discharge of wastewater from a hospital and a pharmaceutical plant was investigated by using Acinetobacter species as environmental bacterial indicators. The level of susceptibility to six antimicrobial agents was determined in 385 Acinetobacter strains isolated from samples collected upstream and downstream from the discharge points of the hospital and the pharmaceutical plant. Results indicated that while the hospital waste effluent affected only the prevalence of oxytetracycline resistance, the discharge of wastewater from the pharmaceutical plant was associated with an increase in the prevalence of both single- and multiple-antibiotic resistance among Acinetobacter species in the sewers.

Effects of requiring prior authorization for selected antimicrobials: expenditures, susceptibilities, and clinical outcomes.

Antimicrobial control programs are widely used to decrease drug expenditures, but effects on antimicrobial resistance and outcomes for patients are unknown. When a requirement for prior authorization for selected parenteral antimicrobial agents was initiated at our urban, county teaching hospital, total parenteral antimicrobial expenditures decreased by 32%. Susceptibilities to all beta-lactam and quinolone antibiotics increased, with dramatic increased susceptibilities in isolates recovered in intensive care units, increased susceptibilities in isolates recovered in other inpatient sites, and little change in susceptibilities in isolates recovered in outpatient sites despite no change in infection control practices. For patients with bacteremia due to gram-negative organisms, overall survival did not change with restrictions. No differences occurred in the median time from initial positive blood culture to receipt of an appropriate antibiotic or in the median time from positive blood culture to discharge from the hospital. Thus, requiring preapproval for selected parenteral agents can decrease antimicrobial expenditures and improve susceptibilities to antibiotics without compromising patient outcomes or length of hospital stay.