Automated Blood Culture Research Articles

Bacterial Profile and Antimicrobial Susceptibility Pattern in Septicemia Suspected Children and Available Treatment Options

OBJECTIVES Septicaemia (blood poisoning) is one of the chief sources of global morbidity and mortality in pediatric patients and presents with multidrug and extensive drug resistance. This study aims to detect the major causative agents, antimicrobial susceptibility patterns and associated factors of bacteraemia among pediatric patients.METHODOLOGY This retrospective cross-sectional study was done at National Medical Center, Karachi microbiology laboratory. Clinical specimens consist of blood. Blood samples were processed in BACTEC's automated blood culture system, and positive samples were sub-cultured on blood, Mac-Conkey. Final identification was done by API 20E and API 20NE (Biomerieux) and confirmed by MicroScan (Beckman coulter)®. The antimicrobial susceptibility was performed by using Bauer– Kirby disk diffusion method.RESULTS A total of 395 pediatric patients were taken in the study. Out of these patients, 226 (57.2 %) were female. The children with age 1-4 years were highly affected. Almost 50% were handled in emergency and transferred for admission. 36.2% of patients were exposed to intravenous devices. 89 (22.5%) patients had a history of prolonged use of antibiotics. Most patients with pneumonia presented with septicemia as a complication, and were detected by Klebsiella pneumonia (35.4%), Acinetobacter bauminii (25.3%) and Pseudomonas aeruginosa (20.3%). CONCLUSION The study revealed that gram-negative organisms are the predominant causative organisms of septicaemia. Antibiotic resistance to Carbapenem is gradually increased in the case of Acinetobacter bauminii and Escherichia coli. These issues can be overcome by early detection of microorganisms and establishing antibiotic stewardship.

A Comparative Study of Conventional and Automated Blood Culture System in Adult Patients

Introduction: Blood culture is considered as the gold standard for the diagnosis of bloodstream infection. Conventional blood culture system is less sensitive and takes longer duration for the detection of bloodstream infections whereas automated blood culture system is more sensitive and rapid in detecting causative organisms of bloodstream infections. This prospective study was undertaken to compare the automated blood culture system with the conventional blood culture system for the identification of microbial pathogens in bloodstream infections. Method: This prospective study was done in Department of Microbiology, Silchar Medical College & Hospital, Silchar for a period of 7 months from November 2021 to May 2022. Blood samples from the patients were inoculated into BD BACTEC Plus Aerobic culture vials for automated blood culture system and Brain Heart Infusion broth for conventional blood culture system. Positive bottles flagged by the automated machine were isolated and identified by doing routine subcultures on Blood agar and MacConkey agar and necessary biochemical tests. The conventional blood culture bottles were processed as per standard protocols. Result: Out of 123 samples, 21(17.09%) showed culture positivity by automated blood culture method and 15(12.19%) showed culture positivity by conventional blood culture method. The most common isolate was Staphylococcus aureus followed by Escherichia coli in automated method whereas the most common isolate in conventional method was Staphylococcus aureus followed by Klebsiella pneumoniae. Conclusion: This study concluded that automated blood culture method is more sensitive than conventional method and detects the presence of microorganisms rapidly causing bloodstream infections. Keywords: Bloodstream infection, Automated blood culture, Conventional blood culture

Diagnosis of Enteric Fever Cases: Current Challenges of Azithromycin Resistant R717Q Gene in Rajshahi, Bangladesh

Salmonella enterica serovars typhi is a contributing typhoid agent, leading to untreatable infections based upon the rising prevalence of antimicrobial resistance in the said agent. Consequently, the main concern of the health service providers to treat expansively drug-resistant (XDR) enteric fever has been dependent on the only remaining oral drug, Azithromycin, for creating emerging resistance against it. Therefore, a cross-sectional descriptive study was conducted in the Microbiology Department of Rajshahi Medical College, Rajshahi, to identify Salmonella species by Fastidious Antibiotic Neutralization (FAN) method in suspected enteric fever cases along with its antimicrobial susceptibility pattern and molecular detection of Azithromycin resistant R717Q gene. Samples were collected purposively. Regarding the population's age distribution, out of 127 specimens, the predominant 37(29.1%) were found within the age group of 35-44 years. Automated blood culture was found positive in 64(50.39%) population; out of them, Salmonella spp were 38(59.37%) of the population. Among them 29(76.31%) have travel history, took street food 25(65.7%), used tap water for drinking 21(55.2%) and had kancha latrine 24(63.1%). It was found that 18.43% were sensitive and 81.57% resistant to Azithromycin. Azithromycin-resistant Salmonella spp. were detected in 31(81.57%), and among Azithromycin resistant R717Q gene was identified in 9(29.03%). The azithromycin-resistant R717Q gene was identified by conventional PCR. Because of its oral course of administration, Azithromycin is considered the widely used drug in Bangladesh, which is used once on a dose basis a day. In order to contribute to the development of this drug resistance, there could be many reasons, and among them there are the availability over the counter and the imperfectly done treatment. Thus, this study would be helpful to formulate a regional hospital empirical therapy by antimicrobial sensitivity and resistance pattern of the gene for Azithromycin resistance in Rajshahi Medical College Hospital.
 TAJ 2022; 35: No-2: 67-73

Control of Candida auris Septicemia in Patients of Trauma Intensive Care Unit in a Tertiary Care Hospital

Introduction: Candida auris is associated with nosocomial outbreaks in intensive care settings, and transmission despite the implementation of enhanced infection prevention and control (IPC) measures is a particular concern. Objective: Isolation and identification of C. auris from suspected cases of fungal septicemia and its surveillance. Materials and Methods: Blood culture samples of all suspected cases of fungal septicemia were taken from the trauma intensive care unit (TICU) of a tertiary care hospital in New Delhi between September 2019 and November 2019 as part of surveillance of device-associated infections. All the samples were obtained by central venous catheter line. Samples were processed in BacT/Alert automated blood culture system. Positive flagged bottles were further subcultured unto blood agar, MacConkey agar, and Sabouraud Dextrose Agar with and without cycloheximide and incubated at 37°C. All suspected Candida isolates were identified by conventional methods and the suspected C. auris isolates were further confirmed by matrix-assisted laser desorption time of flight. Results: Out of 24 suspected fungal septicemia cases from TICU, three patients had C. auris septicemia and all three isolates were resistant to fluconazole and voriconazole. Action Taken: IPC practices were reinforced as per the Centers for Disease Control and Prevention guidelines by the infection control team. Conclusion: This article depicts the emergence of multidrug-resistant C. auris and controlling its spread by appropriate infection control measures.

1541. Impact on Reduction of Blood Culture Incubation Period from Five to Four Days from Large Tertitary Care Center

Abstract Background Hospitals have been experiencing an increasing number of acutely ill patients as well as those requiring prolonged hospitalizations, further worsened by the COVID-19 pandemic. This has resulted in increased orders for blood cultures. Once collected, the culture bottles require an incubation period, typically 5 days. The BD BACTEC Fx, a blood culture incubation automated system, holds blood culture bottles until growth is demonstrated or for 5 days, whichever occurs first. New bottles are placed in the apparatus along with older bottles awaiting growth. Capacity is reached when the machine cannot accommodate new bottles due to limited space. When this occurs, manual plating of blood cultures on agar plates is required. Manual plating adds additional responsibility to already overwhelmed microbiology labs. Methods Following the institutional review board (IRB) approval, data were collected retrospectively at a 765-bed tertiary care center. Data was obtained from positive blood cultures from July 2017 to June 2020. We stratified positive blood cultures based on the day of positivity, from day one to day five. The charts were reviewed for positive blood culture results on day four and day five to evaluate for clinical significance. Clinical significance was determined by an Infectious Disease subspecialty team. Blood culture bottles that were positive on day 4 and day 5 in 1 bottle out of 2 bottles with coagulase-negative species of Staphylococcus, Corynebacterium spp, or Propionibacterium acnes were considered a contaminant and did not warrant further chart review. Results On a retrospective review of data from July 2017 to June 2020, the total number of blood cultures obtained was 120,320. Among them, 7,558 blood cultures were positive. 94% were positive on day 1, 4% positive on day 2, 2% positive on day 3, 1% positive on day 4 and < 1% positive on day 5. Positive blood cultures on day 5 did not have any clinical significance, according to the chart review. Conclusion Our study demonstrated that four days of incubation is sufficient for the BD BACTEC Fx automated blood culture incubation system without compromising clinical significance. Implementation of a 4-day incubation period will help with reducing saturation of slots and will improve readiness to accommodate more blood culture bottles. Disclosures All Authors: No reported disclosures.

900. Evaluation of automated blood culture results and subsequent pharmacist intervention on time to targeted therapy in patients with bacteremia

Abstract Background Bloodstream infections (BSIs) are associated with a high mortality rate and timely administration of empiric broad-spectrum antibiotics is critical. However, prolonged exposure increases the risk of colonization and subsequent infection with resistant bacteria. The use of rapid diagnostics and prospective audit and feedback allows for decreased time to targeted therapy and development of resistance. Methods This quasi-experimental study aimed to evaluate changes in time to targeted antibiotic therapy before and after implementation of a real-time pharmacist notification for bacteremic patients. The pre- and post- intervention periods were 9/1/19–12/31/19 and 9/1/21– 12/31/21, respectively. Bacteremic patients that were identified via automated Bruker’s MBT Sepsityper IVD kit reports and admitted for ≥48 hours were included in the study. Patients were excluded if they had ≥ 1 BSI or polymicrobial BSI during hospitalization, treatment of a concurrent infection, or transitioned to hospice. The primary outcome was change in time to targeted antibiotic therapy. Secondary outcomes included change in broad-spectrum antibiotic days of therapy, total duration therapy, hospital length of stay, and in-hospital all-cause mortality. Results A total of 132 charts were reviewed, 83 of which were included, with 47 in the pre- and 36 in the post-intervention group. The most common isolates in the study across both groups was E. coli (36.1%), Klebsiella spp. (15.7%), MSSA (15.7%) and MRSA (8.4%). A trend towards decreased time to targeted therapy was seen in the post-intervention group (5.4 hours vs 33.0 hours; p = 0.150). Median duration of anti-MRSA therapy was reduced from 49.4 hours in the pre-intervention group to 27.92 hours in the post-intervention group (p=0.035). There was no significant difference in total duration of therapy, days of anti-pseudomonal therapy, length of stay, or in-hospital all-cause mortality. Of the 22 pharmacist interventions, 18 (81.8%) were accepted. The most common pharmacist intervention was therapy de-escalation (72.7%). Conclusion Automated rapid blood culture diagnostic results used in conjunction with real-time notifications to pharmacists, further decreased time to targeted therapy and significantly reduced duration of anti-MRSA therapy. Disclosures All Authors: No reported disclosures.

The Gram stain: Implication in blood culture reporting.

Bloodstream infections are life-threatening. They are responsible for prolonged hospital stays and high healthcare costs. Clinical microbiology has an essential role to play in its management. Direct Gram-stain reporting from positive blood culture bottles has been found helpful. We aimed to evaluate the use of the automated blood culture system in adjunct to the conventional Gram-stain technique, with a direct antimicrobial susceptibility test on the second day. Concordance was highly satisfactory.

Granulicatella elegans infective endocarditis: A case report

Granulicatella elegans is a rare cause of infective endocarditis, accounting for 1–2% of all cases. It is well recognized that this pathogen can present in association with negative blood cultures. There are higher rates of both relapse and mortality compared with endocarditis caused by other bacteria. Microbiological diagnosis can be especially challenging because many conventional blood culture media lack pyridoxal, which can be found in automated blood culture broths like BACTEC™ or BACT/ALERT® and thus they may require ‘helper’ bacteria to be culture the organism. This current case report describes a 66-year-old male patient with a 10-year history of post-inflammatory combined aortic valve disease (moderate aortic stenosis and mild aortic regurgitation). He presented with a 3-month history of recurrent fever and general deterioration. Despite targeted, prolonged, combined antibiotic treatment with intravenous penicillin and gentamicin, surgery was eventually required. An aortic prosthetic valve implantation was performed with good results. In case of subacute endocarditis, especially when a causative organism proves difficult to detect, G. elegans should be considered. Identification is greatly enhanced by using polymerase chain reaction methods and this test should be considered in all cases of culture negative endocarditis.

Evaluation of the relevance of use of the BD-BACTEC®MycosisIC/F, BD-BACTEC®PlusAerobic/F, BD-BACTEC®Lytic/10 anaerobic/F and BD-BACTEC®PedsPlus/F culture bottle system for fungemia detection: A 4-year retrospective study at the Dijon university hospital, France.

Fungemia is a severe invasive fungal infection that combines rapid progression and a high mortality rate. This type of infection is a vital emergency, and early diagnosis is crucial. Currently, only the BD-BACTEC® Automated Blood Culture System (Becton Dickinson, New Jersey, USA) has a medium specifically dedicated to the detection of fungal agents: the BD-BACTEC®MycosisIC/F bottle. Thus, it is important to assess the performance of the different bottles offered by the BD-BACTEC® Automated Blood Culture System for the diagnosis of fungemia. The aim of this study was to evaluate the performance of the BD-BACTEC® MycosisIC/F culture medium in comparison to bacteriologic culture bottle media for the detection of fungemia in different clinical situations. This retrospective study was conducted over a period of 4 years at the Dijon University Hospital. Three hundred and thirty-one pairs of blood cultures (i.e. a BD-BACTEC® MycosisIC/F culture bottle associated with at least one bacteriologic culture media bottle) were included in this study. We showed that the BD-BACTEC® MycosisIC/F culture bottles performed significantly better (i.e. diagnostic advantage either because it was the only positive bottle of the pair or time to positive result was shorter) than the bacteriological culture bottles in 57.7% (191/331) of cases (p <0.01). Multivariate analysis revealed that BD-BACTEC®MycosisIC/F bottles had better diagnostic performance than BD-BACTEC®Bacteriologic bottles in the context of: (i) the initial versus follow-up diagnostic subgroup, (ii) venipuncture or arterial sampling versus other sampling methods, and (iii) detection of filamentous versus yeast fungi. We concluded that the use of BD-BACTEC® MycosisIC/F culture bottles is a relevant addition to media optimized for routine bacterial detection.

Brucellosis in pyrexia of unknown origin: Reminiscing the forgotten entity

PurposeBrucellosis is one of the substantial yet forgotten entity of pyrexia of unknown origin, transmitted via contact with animals or animal products. Despite being an important etiological agent of PUO, it is under-reported owing to its non-specific and protean clinical presentations. Thereby, the present study intended to describe clinical profile and outcome of patients of brucellosis from a tertiary care center in North India. MethodsThis was a retrospective study and included all the patients that were reported positive for Brucella in automated blood culture system (BACTEC) and identified using MALDI-TOF MS over a period of 6 years (2014–2020). ResultsA total of nine cases were reported positive for Brucella during the study period. Fever was noted to be the most common symptom present in all the patients (PUO), followed by fatigue. The patients were managed with monotherapy using doxycycline or combination of cotrimoxazole, rifampicin and doxycycline. All the patients responded well to treatment. ConclusionThe present study highlights the significance of automated blood culture system in conjunction with MALDI-TOF MS for early and reliable identification of Brucella. In the present study, all cultures were reported positive within five days of incubation in automated blood culture system, underlining the significance of using the same which lessens the turnaround time and increases the sensitivity. Owing to the protean manifestations and low sensitivity of the available tests, the clinician needs to have a high level of suspicion to precisely diagnose the illness to prevent undue delay in management.

Molecular and Serological Diagnostic Approach to Define the Microbiological Origin of Blood Culture-Negative Infective Endocarditis.

Blood culture-negative infective endocarditis (BCNIE) poses a significant challenge in determining the best antibiotic regimen for this life-threatening infection, which should be treated with as specific and effective a regimen as feasible. The goal of this study was to determine the prevalence of BCNIE among definite infective endocarditis (IE) cases and to study the impact of a molecular and serological diagnostic approach in defining the microbiological origin of BCNIE. This study included 94 definite IE cases. Serum and blood samples from BCNIE patients were tested using serological, broad-range PCR, and sequencing assays. Valve tissue sections obtained from 42 operated patients were subjected to culture and molecular studies. BCNIE accounted for 63 (67%) of the cases. Of these cases, blood PCR followed by sequencing could diagnose 11 cases. Zoonotic infective endocarditis was detected in 7 (11%) patients by serology and PCR (four Brucella, two Bartonella, and one Coxiella). Sequencing of valve PCR bands revealed 30 positive cases. Therefore, the percentage of BCNIE with unidentified etiology was reduced from 67% to 27.7% through a combination of all diagnostic procedures utilized in our study. Blood and valve PCR and sequencing assays are valuable techniques for the etiological diagnosis of BCNIE, especially in cases with previous antibiotic therapy. However, these tests should be used as part of a larger diagnostic strategy that includes serology, microscopy, and valve culture. The use of an automated blood culture system, and proper blood culture collection before ordering antibiotics, will guide IE etiological diagnosis.

Evaluation of Microbiologically Documented Bloodstream Infections in Pediatric Hematology/Oncology Patients: Results of Ten Years

Inpediatric hematology/oncology patients, infections are the main cause of prolonged hospital stay, increased mortality and high cost following relapse or progression. In this patient group, infections caused by multi-drug resistant bacteria are common and affect morbidity and mortality rates. We aimed to determine the frequency and antibiotic susceptibility of bacteria isolated from blood cultures of the patients with malignant and non-malignant diseases in our hospital over a ten-year period. patients admitted to the Pediatric Hematology/Oncology Service between January 2011- June 2021 were evaluated. The most common disease was acute lymphoblastic leukemia (27%). The first isolated bacteria of same species for each patient were included, contaminated cultures were not included. Blood cultures incubated in the Bactec FX automated blood culture system for five days. Bacteria were identified by conventional methods or automated systems. Antibiotic susceptibility tests were performed by disc diffusion or gradient test and were evaluated according to guidelines. A total of 4631 blood culture samples from 296 patients were analyzed. Positive signal was seen in 620 samples. Blood culture posivity was 13.4%. Total 298 blood culture samples were evaluated. Gram positive bacteria rate were 59% and 41% gram negative. The most frequently (58.7%) isolated gram positive bacteria were methicillin-resistant coagulase negative staphylococci and gram negative bacteria were Klebsiella pneumoniae (28,5%). The rate of bacteria producing extended spectrum beta lactamase (ESBL) was detected as 74% for Escherichia coli and 69% for Klebsiella pneumoniae . It is important for each center to determine its own causative agents and their resistance patterns in bloodstream infections. Gram positive bacteria were found dominantly in our study. The high ESBL rate in E.coli and K.pneumoniae isolates is remarkable. Early detection of the causative agents in bloodstream infections of the pediatric hematology/oncology patients and initiation of prompt treatment are important to reduce mortality.

A diagnostic test of real-time PCR detection in the diagnosis of clinical bloodstream infection

Blood culture remains the standard for diagnosing bloodstream infections, but it is difficult to identify bacteria directly and timeliness. The real-time polymerase chain reaction (PCR) has the potential to fill this diagnostic gap. This study intends to explore the sensitivity and specificity of PCR in detecting bloodstream infection pathogens and to compare it with routine blood culture to explore its clinical application value. A total of 126 patients with bloodstream infections collected from various clinical departments of The First Hospital of Hebei Medical University. The patient's sample was divided into two parts. The one for multiplex PCR detection was performed using the Pathogeno Elite Multiplex PCR kit. Another blood culture was a fully automatic blood culture system from Autobio company. Among the 126 patients, a total of 17 pathogens were detected by PCR and blood culture both methods. PCR detected a total of 43 positive samples and 83 negative samples. Five samples were positive with blood culture, and 81 were negative. The negative predictive value of PCR was 0.98, with a sensitivity of 0.71 and a specificity of 0.68. A total of 38 specimens were positive for PCR but negative for blood culture, and 2 samples were positive for blood culture but negative for PCR. The top 5 pathogens with PCR detection were Epstein-Barr virus (27 cases), Human herpes virus 5 (9 cases), Klebsiella pneumoniae (5 cases), Staphylococcus (5 cases), and Stenotrophomonas maltophilia (4 cases). PCR detection can rapidly identify more pathogens and even multi-pathogen infections. Therefore, PCR testing may improve pathogen detection in patients with suspected bloodstream infections, enabling targeted treatment of patients.

Clinical utility of time to positivity of blood cultures in cases of fungaemia: A prospective study

PurposeFungaemia due to yeast is a major cause of morbidity and mortality in critically ill patients. Although, automated blood cultures have improved the time to diagnosis, very few studies have systematically evaluated the utility of blood culture time to positivity (TTP) of fungaemia in the clinical scenario. In this study, we evaluated the TTP for different yeast species to determine its clinical utility. Material and methodsA prospective study including 244 consecutive patients admitted to the adult (n ​= ​76) and paediatric (n ​= ​168) intensive care units (ICUs) was conducted between December 2017 through March 2019. The clinical and demographic characteristics, BACTEC blood culture results and TTP for yeast positive blood cultures were recorded for analysis. ResultsA total of 244 patients with 357 episodes of candidaemia were enrolled during the study period. The TTP (mean ​± ​SD) for all yeast species was 26.8 ​± ​23.6 ​h while it was significantly longer in paediatric than adult patients (30.5 ± 24.7 vs. 25.2 ​± ​22.9 ​h; p ​= ​<0.0001). Wickerhamomyces anomalus and Cyberlindnera jadinii (previously C. utilis) were exclusively isolated from paediatric population where W. anomalus demonstrated significantly longer TTP than C. jadinii. Among adult cases, C. albicans exhibited significantly longer TTP than C. tropicalis. In paediatric cases, >80% of C. tropicalis and C. utilis flagged positive in blood culture before 24 ​h while majority (65.9%) of W. anomalus isolates flagged positive later than 24 ​h. Similarly in adult samples, 63% of C. tropicalis isolates beeped positive before 24 ​h. ConclusionTTP for yeast may provide insight regarding the responsible yeast species before final identification among critical patients with candidaemia. Larger studies are warranted for evaluating clinical utility of TTP considering other complex factors like yeast burden, generation time, virulence and host factors, which may affect TTP. Keywords: fungaemia; time to positivity; candidaemia; paediatric; adult.

P406 Rise in HIV negative Cryptococcal infection in liver disease patient: epidemiology, risk factor, antifungal susceptibility profile from tertiary care hepatobiliary center

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PMObjective Cryptococcus neoformans is an encapsulated yeast found ubiquitously in the environment, considered important in immunocompromised individuals with HIV. Non-HIV susceptible groups include malignancies, long-term use of corticosteroids, solid organ transplantation, sarcoidosis, immunosuppressive therapy, and cirrhosis. Cryptococcal infections are rising in HIV-negative cirrhotic patients, accounting for 6-21% of the systemic infections. Chronic liver disease (CLD) is a neglected risk factor for fungal infections. To analyze epidemiology, risk factors, and antifungal susceptibility profile of HIV-negative liver disease patients.Material MethodsThe retrospective study was conducted in tertiary care hepatobiliary center in New Delhi, India from January 2017 to March 2022 after ethical approval. Demographic data, laboratory data, and clinical history of the patients admitted with liver disease and suspected cryptococcal infections were obtained from Hospital records for epidemiology and risk factor analysis. Blood, CSF, serum, urine, and body fluid (Ascitic fluid, peritoneal fluid, pleural fluid, MiniBAL) samples were processed for fungal culture on SDA and SDA with cyclohexamide. Blood and body fluid were also inoculated in an automated blood culture system (BacT/ALERT 3D bioMérieux). Samples were processed for gram stain, India ink, and Cryptococcal antigen. Identification was done by an automated identification system (VITEK 2 compact bioMérieux). The micro-broth dilution method (Sensititre YeastOne colorimetric plate, Thermo Fisher Scientific, MA, United States) was used to determine the susceptibility of all Cryptococcus strains to the six antifungal drugs, fluconazole, 5-fluorocytosine, amphotericin B, itraconazole, posaconazole, and voriconazole. The results were reported as wild-type (WT) or non-wild-type (non-WT) in accordance with the epidemiological cutoff value (ECV) set for Cryptococcus spp. (Espinel-Ingroff et al., 2012a, b; CLSI, 2018).ResultsWe analyzed 30 patients of suspected cryptococcal infection and obtained 40 C. neoformans isolates from different samples from these patients. Out of 40 samples, C neoformans was isolated from blood (25,62.5%), urine (6,15%), ascitic fluid (4,10%), mini-BAL (2,5%), bone marrow (1,2.5%), CSF (1,2.5%), pleural fluid (1,2.5%). All samples were positive for Cryptococcal antigen. India ink positivity was observed in 56%. Alcoholic liver disease was the most common risk predictive factor seen in 30% of patients. Hepatitis B and C-associated with CLD was seen in 20%. Other risk predictive factors were AKI (70%), diabetes mellitus (20%), TB (10%), autoimmune hepatitis (6.6%), autoimmune disease (autoimmune hemolytic anemia, Sjogren syndrome) (6.6%), Sarcoidosis (3.3%), Hepatocellular carcinoma (3.3%), HIV (3.3%). Hepatic encephalopathy was seen in 70% of patients which mimics the symptoms of C. meningitis.7.5% (3/40), 5% (2/40), 2.5% (1/40), 7.5% (3/40), and 2.5% (1/40) of C. neoformans strains were non-WT to fluconazole, 5-fluorocytosine, amphotericin B, posaconazole, and itraconazole, respectively, but all strains were WT to voriconazole. Overall mortality was 66.6%.ConclusionAbove study shows that liver diseases are an important risk factor for cryptococcal infection owing to immunosuppressed state, use of steroids, and associated comorbidities like DM, TB, AKI, autoimmune disease, Sarcoidosis, hepatocellular carcinoma. Therefore, it is necessary to investigate and be vigilant about the isolation of Cryptococcus. The use of appropriate antifungals can improve clinical outcomes. As there is increasing resistance to amphotericin B, 5-fluorocytosine, azoles; antifungal susceptibility testing should be done. Cryptococcal antigen detection can be useful in non-CSF samples as well.

P223 First case of Candida auris candidemia in Manipur, Northeast India

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PMObjective Candida auris is known as an emerging ‘superbug’ because of its intrinsic resistance to one or more, sometimes to all available antifungal drugs and spreading globally. It has the ability to cause devastating nosocomial infections. In India, C. auris infection is on the rise with reports from north, south, central and eastern India. Here we present the first case of C. auris fungemia from a tertiary care hospital of Manipur in Northeast India.MethodsA 15-year-old Muslim girl was referred from a private hospital to Regional Institute of Medical Sciences (RIMS) hospital on November 19, 2021 with a history of burning epigastrium, headache, loss of appetite, shortness of breath, dry cough, fever, and generalized weakness for last 3 days. At the time of admission she was cyanotic. Family gave history of congenital heart disease and frequent visits to hospital. Echocardiogram revealed congenital cyanotic heart disease (Tetralogy of Fallot) showing large perimembranous VSD with bidirectional shunt. A complete hemogram showed neutrophilic leukocytosis with shift to left with band form, absolute monocytosis, and increased RBC count with mild anisocytosis. On November 24, 2021, 5 days after admission, her condition deteriorated and she was shifted to ICU. However, the condition of the patient deteriorated and she died on November 29, 2021 due to acute decompensated heart failure. Follow-up of other patients admitted in the same ward revealed no candidemia in next the few weeks.ResultsA single blood culture sent on November 29, 2021 was incubated in an automated blood culture system, BacT Alert and showed growth of budding yeast cells. Growth in SDA revealed it to be Candida sps. and Gram-stained smear examination revealed presence of budding yeast cells but no pseudohyphae. Germ tube test was negative. On CHROM agar, it produces pale yellow colonies at 24 h which progresses to light purple colonies around the rim at 48 h. Further processing in VITEK 2 (Biomerieux) identified it as C. auris. The isolate was sent to National Culture Collection of Pathogenic Fungi, WHO collaborating center, PGIMER and the isolate was confirmed as Candida auris by MALDI-TOF assay.Conclusion Candida auris is spreading irrespective of the level of health care. Blood culture before administration of antibiotics and in febrile sick patients cannot be underestimated. Rapid and accurate identification methods for timely diagnosis and stringent infection control measures with an emphasis on hand hygiene are important to prevent and control C. auris outbreaks.

P049 Candidemia: Isolate profiling and antifungal susceptibility testing experience from Jodhpur, Western India

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM ObjectivesThe study was undertaken over a 9-month study period at a tertiary care and super specialty hospital situated in Jodhpur, Western Rajasthan, India, with the following objectives:To determine the prevalence of Candidemia among all blood culture positivesIsolate profiling or speciation of Candida spp.Antifungal susceptibility testing of the Candida isolatesMethodsAutomated blood culture bottles (BD BACTEC 960) that flagged positive were taken up for gram staining. Those bottles which showed gram-positive budding yeast with or without pseudohyphae were selected as the study isolates for candidemia. All such bottles were subcultures on Sabouraud's dextrose agar and incubates aerobically at 37°C for 2-5 days. Creamy, pasty, off-white colonies of Candida were further taken up for identification by germ tube testing, CHROM agar, and VITEK-MS.Antifungal susceptibility testing was performed for all isolates by VITEK 2 against fluconazole, caspofungin, voriconazole, micafungin, flucytosine, and amphotericin B.ResultsDuring the study period May 1, 2021-January 31, 2022, the microbiology laboratory received a total of 10 841 automated blood culture bottles, of which, overall, 1051 flagged positive. Budding yeasts were seen in 92 bottles.The prevalence of candidemia was found to be 1.49%. Budding yeasts made up 8.75% of all positive blood cultures.Conventional and automated identification methods showed the non-albicans Candida made up the majority (85.87%) of isolates. Candida tropicalis (43.47%) was the most common species overall, followed by C. parapsilosis (17.37%), C. albicans (14.13%), C. guilliermondi (5.43%), C. glabrata (5.43%), and C. auris (4.34%). Two isolates each of C. kruzei, C. utilis, C. rugosa, and Trichosporon spp. were also obtained.The antifungal susceptibility testing results for the commonest species C. tropicalis showed susceptibility of 90% against caspofungin and micafungin, 82% against fluconazole and voriconazole, 45% for flucytosine, and 47.5% against amphotericin B. C. albicans showed 100% susceptibility to fluconazole, and caspofungin, while C. parapsilosis showed a lower susceptibility percentage against all drugs in the panel. The two strains of C. auris were solely susceptible to caspofungin.Demography of the patients showed a male preponderance (M:F ratio was 2:1). The mean age of patients was 44 years.ConclusionThe prevalence of candidemia in Jodhpur, Western India was found to be 1.45%, a figure much less than that reported from most other tertiary care centers of the country. The commonest isolate was C. tropicalis (43.47%), same as that reported from most Indian studies. Our isolates were largely (>90%) susceptible to the drug of choice caspofungin, including the multidrug-resistant C. auris strains. The study findings reflect a low prevalence of candidemia, indicating adequate antibiotic and antifungal stewardship practices at Jodhpur.

Impact of Automated Blood Culture Systems on the Management of Bloodstream Infections: Results from a Crossover Diagnostic Clinical Trial.

ABSTRACTBloodstream infections are associated with high rates of morbidity and mortality. Blood culture remains the gold standard for the diagnosis of BSIs. We report a prospective crossover diagnostic clinical trial comparing the performances of two blood culture incubation systems: Virtuo and Bactec FX. The primary outcome was the time to detection (TTD) (from the loading of the sample into the incubator to the positivity signal). Patients over 16 years old suspected of having bacteremia/fungemia were included. They were divided into two strata with a total of 9,957 blood extractions. Initially, each stratum was randomly assigned to one of the incubators and then alternated every 2 weeks for 6 months. Each sample was inoculated into an aerobic bottle and an anaerobic bottle. All bottles were processed equally according to the laboratory’s standard procedures after they were flagged positive. We analyzed 4,797 samples in the Virtuo system and 5,160 in the Bactec FX system. The median TTD was significantly lower for the Virtuo group (Virtuo, 15.2 h; Bactec FX, 16.3 h [P < 0.0001]). The turnaround time (TAT) (from sample loading to the Gram stain report) was also reduced with Virtuo (Virtuo, 26.2 h; Bactec FX, 28.3 h [P < 0.004]). When considering only samples from patients with antimicrobial treatment prior to blood culture extraction, the TTD was shorter for Virtuo (median differences in the TTD of 4.5 h for all bottles and 8.7 h for aerobic bottles only [P = 0.0001]). In conclusion, virtuo provided shorter TTD and TAT than Bactec FX. The difference in the median TTD was increased when considering samples incubated in aerobic bottles from patients with antimicrobial treatment. This could have an important effect on the faster diagnosis of BSIs.IMPORTANCE Bloodstream infections are associated with high rates of morbidity and mortality. Blood culture remains the gold standard for its diagnosis. While the identification of the pathogen and its antibiotic susceptibility is required to confirm the optimal antimicrobial regimen, reductions in the times to the detection of positivity and reporting of Gram stain results may be important and time-saving to reduce inappropriate antimicrobial use, improve patient outcomes, and decrease health care costs. We report the first clinical diagnostic study of this scale in a “real-world” setting with a crossover design, comparing two automatic blood culture incubators using samples from patients with a suspected diagnosis of bacteremia/sepsis, as opposed to spiked vials. Our study design mimics that of clinical trials performed for drug marketing authorization, but patient randomization was replaced with the crossover design. A shorter time to detection could have an important effect on the faster identification of causative microorganisms of BSIs and antimicrobial stewardship.

POS-034 MICROBIOLOGICAL PROFILE OF CYST INFECTION IN AUTOSOMAL DOMINANT POLYCYSTIC KIDNEY DISEASE IN NORTH INDIA

Autosomal dominant polycystic kidney disease (ADPKD) is most common inherited kidney disorder. Cyst infection is common in ADPKD and could be fatal in some cases. There is limited evidence on management of these case and most cases are treated with empirical antibiotics. We conducted this study to identify the causative microorganisms and determine antibiotic susceptibility to these organisms in our region. This study was conducted between May 2015 to January 2022 at a tertiary care center in Rajasthan. Suwabe criteria was applied to diagnose cyst infection. Abdominal computed tomography (CT) was done in all ADPKD patients with clinical and laboratory features suggestive of cyst infection. Non contrast CT was performed in cases with renal dysfunction. Ultrasound guided percutaneous needle aspiration of infected renal cyst was performed in cases with persistent fever for more than 1 week despite empirical antibiotic therapy. Cyst aspirate was subjected to culture and sensitivity to conventional lipid soluble antibiotics (Fluoroquinolones/ trimethoprim-sulfamethoxazole) using BD BACTEC™ automated blood culture systems (Figure 1). Twenty-seven patients with features suggestive of cyst infection on CT abdomen were subjected to ultrasound guided cyst aspiration. Clinical characteristics of the patients has been depicted in Figure 2. Simultaneous pyuria was present in five (18.5%) cases while urine culture was positive only in two (7.4%) cases. Cyst aspirate culture was positive in eighteen (66.7%) cases. Microbiological profile has been depicted in Figure 3. Escherichia coli was the most common isolated organism. There was low sensitivity to conventional lipid soluble antibiotics with 38.9% (7/18) patients sensitive to fluoroquinolones (Ciprofloxacin/ Levofloxacin) and 50% (9/18) patients sensitive to trimethoprim-sulfamethoxazole. Gram negative bacteria are the most common cause of cyst infection in ADPKD patients. There is high resistance to conventional lipid soluble antibiotics in our region and we should reconsider their empirical use in these patients. We also want to reiterate that cyst aspiration should be considered early as diagnostic modality to find out the causative organisms and their drug sensitivity.

Blood Culture Headspace Gas Analysis Enables Early Detection of Escherichia coli Bacteremia in an Animal Model of Sepsis.

(1) Background: Automated blood culture headspace analysis for the detection of volatile organic compounds of microbial origin (mVOC) could be a non-invasive method for bedside rapid pathogen identification. We investigated whether analyzing the gaseous headspace of blood culture (BC) bottles through gas chromatography-ion mobility spectrometry (GC-IMS) enables differentiation of infected and non-infected; (2) Methods: BC were gained out of a rabbit model, with sepsis induced by intravenous administration of E. coli (EC group; n = 6) and control group (n = 6) receiving sterile LB medium intravenously. After 10 h, a pair of blood cultures was obtained and incubated for 36 h. The headspace from aerobic and anaerobic BC was sampled every two hours using an autosampler and analyzed using a GC-IMS device. MALDI-TOF MS was performed to confirm or exclude microbial growth in BCs; (3) Results: Signal intensities (SI) of 113 mVOC peak regions were statistically analyzed. In 24 regions, the SI trends differed between the groups and were considered to be useful for differentiation. The principal component analysis showed differentiation between EC and control group after 6 h, with 62.2% of the data variance described by the principal components 1 and 2. Single peak regions, for example peak region P_15, show significant SI differences after 6 h in the anaerobic environment (p < 0.001) and after 8 h in the aerobic environment (p < 0.001); (4) Conclusions: The results are promising and warrant further evaluation in studies with an extended microbial panel and indications concerning its transferability to human samples.