Comparison of ciprofloxacin, cotrimoxazole, and doxycycline on Klebsiella pneumoniae: Time-kill curve analysis.

Background: Antibiotic resistance is a significant problem in the world, so optimization of antibiotic use is needed. Klebsiella pneumoniae is a Gram-negative bacterium that causes bacteremia, sepsis, UTIs, pneumonia, nosocomial infections and ESBL-producing bacterium. ciprofloxacin, cotrimoxazole, and doxycycline are broad-spectrum antibiotics, including in WHO essential drugs. Objective: The study tested antibiotics that most effectively inhibited Klebsiella pneumoniae non-ESBL, Klebsiella pneumoniae ESBL invitro with time-kill curve analysis. Method: This experiment used Klebsiella pneumoniae ATCC isolates, stored clinical isolates of Klebsiella pneumoniae non-ESBL, Klebsiella pneumoniae ESBL, and the control group. Isolates other than control were challenged with ciprofloxacin, cotrimoxazole, and doxycycline oral preparations with concentrations of 1, 2, 4 MIC at 0, 2, 4, 6, 8, 24 h. At each hour, the bacteria were cultured, incubated, calculated the number of colonies. The results were analyzed with time-kill curve and tested statistics. Statistical analysis used included ANOVA, post-Hoc, Mann Whitney, and Kruskal Willis tests with p < 0.05. Results: Ciprofloxacin, cotrimoxazole, and doxycycline in this study had inhibition effects on Klebsiella pneumoniae non-ESBL and Klebsiella pneumoniae ESBL. Ciprofloxacin had the best inhibitory effect. Statistically, the most meaningful differences of antibiotics in ciprofloxacin and cotrimoxazole at four and 24 h (p < 0.001), in concentrations of 1 MIC and 4 MIC at 2 h (p < 0.001), and in Klebsiella pneumoniae ESBL and Klebsiella pneumoniae ATCC at 8 h (p = 0.024). Conclusion: Ciprofloxacin is the best antibiotic to inhibit the growth of Klebsiella pneumoniae non-ESBL and Klebsiella pneumoniae ESBL compared to cotrimoxazole and doxycycline. The inhibitory effect increases with an increase in concentration.

Evaluation of fluconazole, itraconazole, and voriconazole activity on Candida albicans: A case control study.

Background: Azole antifungals are the most commonly used antifungals. The high use of azoles for long-term therapy and prophylaxis is prone to cause resistance. Thus, it is necessary to evaluate the antifungal activity against Candida albicans. Objectives: Analyzing the comparison of antifungal exposure on the time-kill curve to Candida albicans. Method: A case-control study was conducted with a posttest control group design. This study used Candida albicans clinical and ATCC isolates exposed to antifungal solutions with 1 ×, 4 ×, and 16 × minimum inhibitory concentrations (MIC). Antibiotics used included fluconazole, itraconazole, and voriconazole. Candida albicans isolates were incubated with MIC, and the number of colonies was counted at 0, 2, 4, 8, 12, 24, and 48 h. The number of colonies that grew every hour of observation was included in the time-kill curve. The data were then analyzed using an ANOVA test with p <0.05. Results: The antifungals (fluconazole, itraconazole, and voriconazole) showed fungistatic activity against Candida albicans clinical and ATCC isolates. There was a significant comparison between the antifungal group and the control group at 12, 24, and 48 h. The most significant difference between antifungal and control group was found at 24 h where fluconazole had 95% CI = 0.807-2.061 (p <0.001), itraconazole 95% CI = 0.722-1.976 (p <0.001), and voriconazole CI 95% = 0.807-2.062 (p <0.001). Conclusion: Fluconazole, itraconazole, and voriconazole were effective in inhibiting the growth of Candida albicans. Maximum inhibition in vitro occurs after 12 h of antifungal exposure.

Growth kinetics of multiple Acinetobacter baumannii resistotype after meropenem-based antibiotic combination exposure.

Background: Carbapenems are the treatment of choice for multidrug-resistant (MDR) and extensively drug-resistant (XDR)  Acinetobacter baumannii infections, but the emergence of carbapenem-resistant  A. baumannii (CRAB) has rendered it ineffective in the vast majority of cases. Combination therapy has grown in popularity over the last decade; this study aims to analyze  A.baumannii growth kinetics after exposure to meropenem and ampicillin-sulbactam compared with meropenem and amikacin antibiotic combinations in clinically relevant concentrations.  Methods: This experimental laboratory study was conducted on the  A.baumannii ATCC 19606 isolate and three clinical isolates that were intermediate or resistant to tested antibiotics. Meropenem and ampicillin-sulbactam, as well as meropenem and amikacin, were tested at four different concentrations against isolates. Turbidity measurements were taken at predetermined time points of 0, 1, 2, 4, 6, 8, and 24 hours following exposure; bacterial concentration was enumerated using the agar plate method, with the results plotted in a time-kill curve.   Results: A bactericidal effect was achieved in isolates that were intermediate to ampicillin sulbactam and resistant to meropenem after the administration of meropenem and ampicillin-sulbactam combination with a concentration of 4 µg/ml and 16/8 µg/ml, respectively. The combination of meropenem and ampicillin-sulbactam demonstrated bacteriostatic activity against isolates that were resistant to both antibiotics. Isolates treated with resistant antibiotics showed an increased growth rate compared to the growth control.  Conclusion: The combination of meropenem and ampicillin-sulbactam could be a promising combination therapy in treating CRAB infections. The mechanism and degree of antibiotic resistance in the isolates affect the efficacy of antibiotic combinations; further research is needed to corroborate the findings of this study.

Effect of Ciprofloxacin, Levofloxacin, and Ofloxacin on Pseudomonas aeruginosa: A case control study with time kill curve analysis.

Background: Antibiotic resistance is closely related to therapy failure. Most antibiotic resistance is caused by delays in determining antibiotic agents, low administration doses, long periods between doses (inadequate pharmacokinetics) and single drug administration in infections caused by more than one pathogen. Treatment of Pseudomonas aeruginosa (P. aeruginosa) with ciprofloxacin, levofloxacin, and ofloxacin as monotherapy can lead to drug resistance, although combination therapy also does not provide a better outcome. Objective: To analyze the time-kill curve for P. aeruginosa and Multidrug resistance (MDR) P. aeruginosa. Methods: This research is a case control study using isolates of P. aeruginosa ATCC 27853, clinical isolates of P. aeruginosa and MDR P. aeruginosa. Exposure of ciprofloxacin, levofloxacin, and ofloxacin to isolates with 1MIC, 2MIC, and 4MIC were then cultured at 0, 2, 4, 6, 8, 24 h of testing, then counting the number of colonies that grew and then analyzed by time-kill curve and statistical tests. The statistical test used in this study was the ANOVA and Mann-Whitney test with p < 0.05. Results: Ciprofloxacin and ofloxacin achieved bactericidal activity, especially at a concentration of 4MIC. Levofloxacin ultimately achieved bactericidal activity at all concentrations. Statistical analysis showed there were significant differences in the number of colonies p < 0.001 in the second, fourth, sixth, and eighth hour between the three isolates, p < 0.001 in the sixth and second 4 h between 1MIC and 4MIC, p = 0.012 in the second 4 h between levofloxacin and ofloxacin antibiotics. Conclusion: Levofloxacin has shown to have better bactericidal activity than ciprofloxacin, and ciprofloxacin has almost the same bactericidal activity as ofloxacin in vitro tests seen from the time-kill curve.

Multicenter Study of the Risk Factors and Outcomes of Bloodstream Infections Caused by Carbapenem-Non-Susceptible Acinetobacter baumannii in Indonesia.

The prevalence of bacteremia caused by carbapenem-non-susceptible Acinetobacter baumannii (CNSAB) continues to increase, and it is associated with a high mortality rate. Early recognition of infection and mortality determinants risk factors is necessary for adequate antibiotic administration. We aimed to determine the risk factors and outcomes of CNSAB bacteremia in Indonesia. A multicenter case-control study was conducted in three referral hospitals in Indonesia. Data were collected retrospectively from January 2019 to December 2021. Cases were defined as patients with bacteremia where CNSAB was isolated from the blood, while the controls were patients with bacteremia caused by carbapenem-susceptible A. baumannii (CSAB). Risk factors for bacteremia and mortality associated with CNSAB bacteremia were determined using univariates analysis (chi-squared and Student's t-test or Mann-Whitney test) and multivariate logistic regression analysis. A total of 144 bacteremia patients were included, of whom 72 patients were for each case and control group. The final model of multivariate regression analysis revealed that bacteremia source from the lower respiratory tract (adjusted odds ratio (aOR): 3.24; 95% CI: 1.58-6.63, p = 0.001) and the use of central venous catheter (aOR: 2.56; 95% CI: 1.27-5.18; p = 0.009) were independent risk factors for CNSAB bacteremia. Charlson Comorbidity Index ≥ 4 (aOR: 28.56; 95% CI: 3.06-265.90, p = 0.003) and Pitt Bacteremia Score ≥ 4 (aOR: 6.44; 95% CI: 1.17-35.38; p = 0.032) were independent risk factors for mortality due to CNSAB bacteremia. Only high Pitt Bacteremia Score was an independent risk factor for mortality of CSAB bacteremia. In conclusion, we identified the risk factors for CNSAB-associated bacteremia and the risk factors for death, which are relevant for empiric therapy and infection control prevention, as well as prognosis evaluation of patients with bloodstream infections.

Distribution of Carbapenemase Genes among Carbapenem-Non-Susceptible Acinetobacter baumanii Blood Isolates in Indonesia: A Multicenter Study.

Carbapenem non-susceptible Acinetobacter baumannii (CNSAB) is an important pathogen that causes nosocomial bacteremia among critically ill patients worldwide. The magnitude of antibiotic resistance of A. baumanii in Indonesia is expected to be significant; however, the data available are limited. The aim of this study was to analyze the genetic profiles of CNSAB isolates from patients with bacteremia in Indonesia. CNSAB isolates from blood cultures of bacteremia patients in 12 hospitals in Indonesia were included. The blood cultures were conducted using the BacT/Alert or BACTEC automated system. The CNSAB were identified with either Vitek 2 system or Phoenix platform followed by a confirmation test using a multiplex polymerase chain reaction (PCR) assay, targeting the specific gyrB gene. The carbapenemase genes were detected by multiplex PCR. In total, 110 CNSAB isolates were collected and were mostly resistant to nearly all antibiotic classes. The majority of CNSAB isolates were susceptible to tigecycline and trimethoprim-sulfamethoxazole (TMP-SMX), 45.5% and 38.2%, respectively. The blaOXA-51-like gene was identified in all CNSAB isolates. Out of the total, 83.6% of CNSAB isolates had blaOXA-23-like gene, 37.3% blaOXA-24-like gene, 4.5% blaNDM-1 gene, 0.9% blaIMP-1 gene, and 0.9% blaVIM gene. No blaOXA-48-like gene was identified. The blaOXA-23-like gene was the predominant gene in all except two hospitals. The presence of the blaOXA-24-like gene was associated with resistance to tigecycline, amikacin, TMP-SMX and cefoperazone-sulbactam, while blaOXA-23-like gene was associated with resistance to TMP-SMX and cefoperazone-sulbactam. In conclusion, the blaOXA-23-like gene was the predominant gene among CNSAB isolates throughout Indonesia. A continuous national surveillance system needs to be established to further monitor the genetic profiles of CNSAB in Indonesia.

Challenge of Ziehl-Neelsen stain for Basidiobolomycosis diagnosis in Indonesia: A unique case report.

BACKGROUND: Basidiobolomycosis is a rare fungal infection in Indonesia, so alternative investigations are needed to make a diagnosis. CASE PRESENTATION: A 24-year-old male patient, Javanese, presented with Basidiobolomycosis. The patient had a lump on the right arm and face 2 years ago, and the tumor progressively grew up and extended to the right upper arm, neck, and face since 1 month ago. This paper discussed the challenge of Ziehl-Neelsen (ZN) stain as a diagnostic tool for Basidiobolomycosis that is a rare case in the world, especially in Indonesia. DISCUSSION: Acid-fast stain has never been used to diagnose fungal cases in general, but it could describe the hyphae and Splendore-Hoeppli very clearly compared to the standard hematoxylin and eosin (HE) stains. CONCLUSION: ZN can be used as an alternative diagnosis of Basidiobolomycosis in a low-resource setting.

The clinical impact of bacterial co-infection among moderate, severe and critically ill COVID-19 patients in the second referral hospital in Surabaya.

Background: Data on the prevalence of bacterial co-infections among COVID-19 patients are limited, especially in our country, Indonesia. We aimed to assess the rate of bacterial co-infections in hospitalized COVID-19 patients and report the most common microorganisms involved and the antibiotic use in these patients. Methods: This study is a retrospective cohort study, among COVID-19 adult patients admitted to Universitas Airlangga Hospital Surabaya from 14 March-30 September 2020. The bacterial infection is defined based on clinical assessment, laboratory parameters, and microbiology results. Results: A total of 218 patients with moderate to critical illness and confirmed COVID-19 were included in this study. Bacterial infection was confirmed in 43 patients (19.7%). COVID-19 patients with bacterial infections had longer hospital length of stay (17.6 ± 6.62 vs 13.31±7.12), a higher proportion of respiratory failure, intensive care treatment, and ventilator use. COVID-19 patients with bacterial infection had a worse prognosis than those without bacterial infection (p<0.04). The empirical antibiotic was given to 75.2% of the patients. Gram-negative bacteria were commonly found as causative agents in this study (n = 39; 70.37%). Conclusion: COVID-19 patients with bacterial infection have a longer length of stay and worse outcomes. Healthcare-associated infections during intensive care treatment for COVID-19 patients must be carefully prevented.

A Randomized, Double-Blind, Multicenter Clinical Study Comparing the Efficacy and Safety of a Drug Combination of Lopinavir/Ritonavir-Azithromycin, Lopinavir/Ritonavir-Doxycycline, and Azithromycin-Hydroxychloroquine for Patients Diagnosed with Mild to Moderate COVID-19 Infections.

BACKGROUND: At the present time, COVID-19 vaccines are at the testing stage, and an effective treatment for COVID-19 incorporating appropriate safety measures remains the most significant obstacle to be overcome. A strategic countermeasure is, therefore, urgently required. AIM: This study aims to evaluate the efficacy and safety of a combination of lopinavir/ritonavir-azithromycin, lopinavir/ritonavir-doxycycline, and azithromycin-hydroxychloroquine used to treat patients with mild to moderate COVID-19 infections. Setting and Design. This study was conducted at four different clinical study sites in Indonesia. The subjects gave informed consent for their participation and were confirmed as being COVID-19-positive by means of an RT-PCR test. The present study constituted a randomized, double-blind, and multicenter clinical study of patients diagnosed with mild to moderate COVID-19 infection. MATERIALS AND METHODS: Six treatment groups participated in this study: a Control group administered with a 500 mg dose of azithromycin; Group A which received a 200/50 mg dose of lopinavir/ritonavir and 500 mg of azithromycin; Group B treated with a 200/50 mg dose of lopinavir/ritonavir and 200 mg of doxycycline; Group C administered with 200 mg of hydroxychloroquine and 500 mg of azithromycin; Group D which received a 400/100 mg dose of lopinavir/ritonavir and 500 mg of azithromycin; and Group E treated with a 400/100 mg dose of lopinavir/ritonavir and 200 mg of doxycycline. RESULTS: 754 subjects participated in this study: 694 patients (92.4%) who presented mild symptoms and 57 patients (7.6%) classified as suffering from a moderate case of COVID-19. On the third day after treatment, 91.7%-99.2% of the subjects in Groups A-E were confirmed negative by a PCR swab test compared to 26.9% in the Control group. Observation of all groups which experienced a significant decrease in virus load between day 1 and day 7 was undertaken. Other markers, such as CRP and IL-6, were significantly lower in all treatment groups (p < 0.05 and p < 0.0001) than in the Control group. Furthermore, IL-10 and TNF-α levels were significantly elevated in all treatment groups (p < 0.0001). The administration of azithromycin to the Control group increased CRP and IL-6 levels, while reduced IL-10 and TNF-α on day 7 (p < 0.0001) compared with day 1. Decreases in ALT and AST levels were observed in all groups (p < 0.0001). There was an increase in creatinine in the serum level of the Control, C, D, and E groups (p < 0.05), whereas the BUN level was elevated in all groups (p < 0.0001). CONCLUSIONS: The study findings suggest that the administration of lopinavir/ritonavir-doxycycline, lopinavir/ritonavir-azithromycin, and azithromycin-hydroxychloroquine as a dual drug combination produced a significantly rapid PCR conversion rate to negative in three-day treatment of mild to moderate COVID-19 cases. Further studies should involve observation of older patients with severe clinical symptoms in order to collate significant amounts of demographic data.