Characteristics, outcome and risk factors for mortality of paediatric patients with ICU-acquired candidemia in India: A multicentre prospective study.

BACKGROUND: The epidemiology, clinical profile and outcome of paediatric candidemia vary considerably by age, healthcare settings and prevalent Candida species. Despite these differences, few comprehensive studies are undertaken. This nationwide study addresses this knowledge gap. METHODS: 487 children who contracted ICU-acquired candidemia at 23 Indian tertiary care centres were assessed for 398 variables spanning demography, clinical characteristics, microbiology, treatment and outcome. RESULTS: Both neonates (5.0 days; range = 3.0-9.5) and non-neonatal children (7.0 days; range = 3.0-13.0) developed candidemia early after ICU admission. Majority of neonates were premature (63.7%) with low birthweight (57.1%). Perinatal asphyxia (7.3%), pneumonia (8.2%), congenital heart disease (8.4%) and invasive procedures were common comorbidities, and antibiotic use (94.1%) was widespread. C tropicalis (24.7%) and C albicans (20.7%) dominated both age groups. Antifungal treatment (66.5%) and removal of central catheters (44.8%) lagged behind. Overall resistance was low; however, emergence of resistant C krusei and C auris needs attention. The 30-day crude mortality was 27.8% (neonates) and 29.4% (non-neonates). Logistic regression identified admission to public sector ICUs (OR = 5.64), mechanical ventilation (OR = 2.82), corticosteroid therapy (OR = 8.89) and antifungal therapy (OR = 0.22) as independent predictors of 30-day crude mortality in neonates. Similarly, admission to public sector ICUs (OR = 3.62), mechanical ventilation (OR = 3.13), exposure to carbapenems (OR = 2.18) and azole antifungal therapy (OR = 0.48) were independent predictors for non-neonates. CONCLUSIONS: Our findings reveal a distinct epidemiology, including early infection with a different spectrum of Candida species, calling for appropriate intervention strategies to reduce candidemia morbidity and mortality. Independent factors identified in our regression models can help tackle these challenges.

Antimicrobial resistance and molecular subtypes of Salmonella enterica serovar Typhi isolates from Kolkata, India over a 15 years period 1998-2012.

Typhoid fever, caused by Salmonella enterica serovar Typhi (S. Typhi), remains an unresolved public health problem in India. Emergence of antimicrobial resistant strains poses a great concern for typhoid treatment and influences reshaping of current S. Typhi population. We included representative S. Typhi strains (n=164) from retrospective studies, both community and hospital based, conducted at National Institute of Cholera and Enteric Diseases, Kolkata during 15 years period (1998-2012) to analyze their antimicrobial resistance (AMR) profiles, mechanism of AMR and molecular subtypes of the strains. More than 60% of the S. Typhi isolates were obtained from community based studies. During the study period, steady decline (46.4%-15.6%) in isolation of multidrug-resistant (MDR, resistant to ampicillin, chloramphenicol and co-trimoxazole) S. Typhi was noticed with parallel increase of nalidixic acid-resistant (NALR) strains (60.7%-93.8%) and ciprofloxacin resistant (CIPR) strains (0%-25%). Of 53 MDR strains, 46 (86.8%) were NALR showing decreased ciprofloxacin susceptible (DCS) (MIC for ciprofloxacin 0.12-0.5µg/ml) phenotype. Conjugative IncHI1 (230kb) and non-conjugative non-IncHI1 (180kb) plasmids were found in 23 (43.4%) and 14 (26.4%) MDR strains respectively, plasmid was absent in 16 (30.2%) MDR strains. MDR strains with or without plasmid shared the same set of resistance genes (blaTEM-1, catA1, sul1, sul2, strA and strB) and class 1 integron possessing dfrA7 gene cassette. Two S. Typhi strains harbored 50kb transferrable plasmids carrying dfrA15 and aadA1 gene cassettes in class 1 integron. The majority of the strains (135/164, 82.3%) belonged to H58 haplotype. Among the MDR isolates, fluoroquinolone resistant or combined resistant isolates (n=147), 127 (86.4%) were H58 and 20 (13.6%) belonged to non-H58. NALRS. Typhi strains with decreased susceptibility or resistance to ciprofloxacin had point mutation(s) in quinolone resistance-determining region of gyrA and parC genes. Pulsed-field gel electrophoresis showed more diversity among NALRS. Typhi than MDR strains. Results of this study generated information useful for better understanding of the disease epidemiology and its control in endemic settings.

Evaluation of fliC-d based direct blood PCR assays for typhoid diagnosis.

BACKGROUND: Typhoid cases need to be diagnosed accurately for early antibiotic therapy and reducing mortality. Identification of Salmonella Typhi (S. Typhi) in blood culture is conclusive, but has poor sensitivity. Detection of S. Typhi by PCR from blood sample has shown promise. Real-time quantitative PCR (Q-PCR) has been widely used in diagnostics for its rapidity and reliability. In the present study, the performance of molecular methods like conventional PCR (C-PCR), nested PCR (N-PCR) and Q-PCR were investigated and compared by targeting S. Typhi specific flagellar fliC-d gene directly in blood samples for typhoid diagnosis. RESULTS: Analytical sensitivities and specificities of the PCR assays were determined under laboratory condition followed by diagnostic performances were demonstrated in 110 clinically diagnosed typhoid fever (CDTF) cases included as study subjects. The DNA detection limit of C-PCR was observed 3 × 10(4) copies/reaction; those of N-PCR and Q-PCR (cutoff Ct value, ≤37) were 3 copies/reaction. The C-PCR was not further evaluated since it showed negative results with all clinical samples due to low sensitivity. Low isolation rate (21.8 %, 24/110) of S. Typhi by blood culture did not reflect the true burden of typhoid fever among the study subjects. Hence diagnostic performances of N-PCR and Q-PCR were determined considering CDTF cases positive by any of the diagnostic assay methods (n = 81) as true positives. Laboratory confirmed non-typhoidal cases (n = 29) were included as true negatives. On comparison, although both the assays were 100 % specific; sensitivity (91.4 % vs. 81.5 %) and efficiency (93.6 % vs. 86.4 %) of Q-PCR were better, but statistically not significant (p > 0.1) than N-PCR. The positive and negative likelihood ratios of Q-PCR were ∞ and 0.09 which indicated the potential clinical utility of Q-PCR for typhoid diagnosis. Q-PCR was more rapid than N-PCR (2 h vs. 6 h) in obtaining test results. CONCLUSIONS: This study demonstrates for the first time that TaqMan-based Q-PCR assay performs more favorably than N-PCR for direct detection of S. Typhi DNA in blood samples. Direct and quantitative blood Q-PCR is a rapid and reliable method for diagnosis of typhoid fever.

Brain abscess due to Cladophialophora bantiana: a review of 124 cases.

Brain abscess caused by Cladophialophora bantiana is a rare disease associated with high mortality due to delay in diagnosis and absence of standardized therapy. We reviewed 124 culture proven C. bantiana brain abscess cases; 103 cases published in English literature during 1952 through 2014 and 21 unpublished cases from our reference center. The majority (57.3%) of the patients was from Asian countries especially from India (62/124, 50%). The diagnosis of the cases was delayed with mean duration 115 days after developing symptoms. The disease was nearly equally distributed in immunocompetent and immunosuppressed hosts but associated with significantly higher mortality (77.1%) in later group. Complete excision of brain lesion in immunocompetent host led to significantly better survival (43.7%). Though all commercially available antifungal drugs have been used in these patients, amphotericin B deoxycholate or lipid preparations were most commonly (62.83%) prescribed agent. None of the drugs used was found to be independently associated with improved outcome. In vitro antifungal susceptibility testing of 13 isolates of our center, demonstrated good activity to voriconazole, posaconazole, and itraconazole, but these triazoles were prescribed in only 29.2% patients. Increased awareness with early suspicion of the disease, and aggressive medical and surgical approach in treating these patients may improve the outcome.

A CLADOPHIALOPHORA BRAIN ABSCESS IN A RENAL TRANSPLANT RECIPIENT.

Cladophialophora bantiana is a dematiceous fungus with neurotrophic propensity for which therapy is not standardized and mortality is high. We report here a 49 year old non-diabetic renal transplant patient on triple immunosuppressant therapy who presented with a history of falls, weakness and headaches. A computed tomography scan of the brain revealed a 30 mm diameter abscess in the brain which was removed surgically and diagnosed on culture as C. bantiana. The patient was successfully treated medically post-operatively with chronic posaconazole. We review the literature regarding central nervous system infections due to C. bantiana.

Melioidosis: Series of Eight Cases.

OBJECTIVE: Melioidosis caused by the Gram-negative bacterium Burkholderia pseudomallei is a very serious infection and has been sporadically reported from the Indian subcontinent. This disease entity can have acute and chronic presentations involving different organ systems. The purpose of this study is to analyze the risk factors, clinical presentations, therapy and outcome of culture proven cases of melioidosis. METHODS: We carried out a retrospective study of eight culture proven cases of melioidosis at a tertiary care hospital in West Bengal. RESULTS: In this series we have found that melioidosis presents with a variety of symptoms ranging from acute presentations in the form of fulminant septicaemia, multiple abscesses in internal organs, osteomyelitis to more chronic form of the infection masquerading as tuberculosis. Bone and joint involvement are particularly common. Diabetes mellitus and chronic alcoholism are significant risk factors. CONCLUSIONS: India or parts of India are possibly endemic areas for melioidosis. Lack of awareness and the diversity of its presentation are probably responsible for underdiagnosis and under reporting.

Paratracheal abscess by plant fungus Chondrostereum purpureum- first case report of human infection.

Chondrostereum purpureum, is a plant fungus causing silver leaf disease of plants, particularly of the rose family. Here we report a case of paratracheal abscess caused by C. purpureum. This is a first of its kind of a case wherein this plant fungus caused disease in a human. Conventional techniques (microscopy and culture) failed to identify the fungus. Only by sequencing, the identity of this unusual pathogen could be revealed. This case highlights the potential of environmental plant fungi to cause disease in humans and stresses the importance of molecular techniques to identify the causative fungal species.

Cytomegalovirus (CMV) as a cause of exacerbation of ulcerative colitis (UC). An approach to diagnosis and management.

Cytomegalovirus (CMV) reactivation may be a cause of acute exacerbations in patients with severe ulcerative colitis (UC). Patients with exacerbation of symptoms by CMV have greater morbidity and mortality rate. The present study was conducted to evaluate the prevalence of CMV infection in UC patients by endoscopic examination, histopathological examination and CMV DNA PCR of colonic mucosal biopsy tissue and also to identify the associated risk factors and clinical outcomes.

A Case of Pacemaker Associated Aspergillus fumigatus Endocarditis.

The use of cardiovascular implantable electronic devices (CIED) is associated with improved quality of life and decreased fatal outcomes in patients with cardiac dysfunctions. As with all foreign devices that are inserted or implanted in the body, CIED also carries the risk of device-related infections. Infections account for <2% of the complications associated with CIED, and only about 2% of these are secondary to a fungal pathogen. The first case of Aspergillus endocarditis secondary to a transvenous pacing lead was reported in the 1980s, and a limited number of cases have been documented in the literature since then. Aspergillus endocarditis is a highly fatal disease and establishing the diagnosis sufficiently early is challenging. We here report a case of Aspergillus endocarditis secondary to permanent pacemaker insertion which was successfully treated following the establishment of the diagnosis using imaging studies and galactomannan assay.

Changes in antimicrobial resistance and molecular attributes of Salmonellae causing enteric fever in Kolkata, India, 2014-2018.

Globally, enteric fever caused by Salmonella Typhi (S. Typhi, ST) and S. Paratyphi A (SPA) remain one of the major diseases of public health importance. In this study, a total of 457 (380 ST, 77 SPA) blood isolates were collected from three tertiary care hospitals in Kolkata during 2014-18. Additionally, 66 (3.4%) ST and 5 (0.25%) SPA were recovered from blood culture of 1962 patients attending OPD of one pediatric hospital during 2016-18. The study isolates were tested for antimicrobial resistance (AMR) profiles; AMR genes; molecular sub-types by PFGE, MLVA and CRISPR. Among the total 446 ST and 82 SPA isolates, fluoroquinolone (FQ) resistance was very common in both serovars. Ciprofloxacin resistance of 24.9% and 9.8% & ofloxacin resistance of 20.9% and 87.8% were found in ST and SPA respectively. Majority (>70%) of the isolates showed decreased susceptibility to ciprofloxacin (DCS). A single point mutation in gyrA gene (S83F) was responsible for causing DCS in 37.5% (n = 42/112) ST and 63% (n = 46/73) SPA isolates. Multidrug resistance (MDR) was found only in 3.4% ST isolates and encoded the genes blaTEM-1, catA, sul, strA-strB, class 1 integron with dfrA7. All MDR ST (n = 15) possessed non-conjugative non-IncHI1 (180 kb) plasmid except one having conjugative IncHI1 (230 kb) plasmid and one without plasmid. The MDR genes were integrated near chromosomal cyaA gene site in ST with/without the presence of plasmid (nonIncH1). Almost 65.7% resistant ST belonged to H58 haplotype. PFGE showed clonally related isolates with 81% similarity in ST and 87% in SPA. Similarly, CRISPR typing showed less diversity among the isolates. However, the isolates (ST and SPA) were found to be more diverse by MLVA typing (D value 0.987 and 0.938). The study reports decrease in MDR and increase in FQ resistance among typhoidal Salmonella isolates over the years giving interesting information for enteric fever treatment.

Septicaemic melioidosis.

Melioidosis is an emerging infectious disease in our country. It is an important cause of community-acquired 'sepsis syndrome' particularly in patients with underlying immunosuppression which often goes undetected due to lack of awareness resulting in high fatalities. Here we report a case of septicaemic melioidosis in a diabetic patient.

Report of Relapse Typhoid Fever Cases from Kolkata, India: Recrudescence or Reinfection?

Three relapse cases were reported out of 107 hospital-attending typhoid cases within a period of 2 years (2014-2016) from Apollo Gleneagles Hospital, Kolkata, India. During the first episode of typhoid fever, 2 of the 3 cases were treated with ceftriaxone (CRO) for 7 days, and 1 was treated for 14 days. Six Salmonella Typhi (S. Typhi) isolates, obtained from the 3 patients during both typhoid episodes, were subjected to antimicrobial susceptibility testing, detection of quinolone resistance-determining region (QRDR) mutation and molecular subtyping by pulsed-field gel electrophoresis (PFGE), multiple-locus variable number tandem repeat analysis (MLVA), multilocus sequence typing (MLST), clustered regularly interspaced short palindromic repeats (CRISPR), and H58 haplotyping. Pairs of the S. Typhi strains isolated from two of the patients during the 1st and 2nd episodes were similar with respect to the antimicrobial resistance (AMR) profiles, QRDR mutations, and molecular subtypes; whereas, the S. Typhi strain pair isolated from the 3rd patient were different in their AMR profiles, QRDR mutations, and MLVA profiles. From these observations, it may be concluded that in spite of treating typhoid cases with CRO for 7-14 days, relapse of typhoid fever might occur. The article also showed the advantage of MLVA typing over PFGE, MLST, and CRISPR typing for the discrimination of strains isolated from the same patient in case of relapse of typhoid fever.

Severe dengue due to secondary hemophagocytic lymphohistiocytosis: A case study.

Dengue, transmitted by the mosquito Aedes aegypti affects millions of people worldwide every year. Dengue induced hemophagocytic lymphohistiocytosis (HLH) is a serious condition and may prove fatal if not detected early and treated appropriately. Diagnosis of HLH is challenging and usually missed as clinical and laboratory findings are nonspecific. Moreover, the pathophysiology of the systemic inflammatory response syndrome and/or sepsis is remarkably similar to HLH. Secondary HLH following infection by the dengue virus is now being increasingly recognized as a cause of severe form of the disease. We report a case of dengue associated HLH in an otherwise healthy person who deteriorated during the course of hospitalization. A disproportionately high ferritin level and persistent bicytopenia prompted investigations for HLH. Diagnosis of dengue fever with virus-associated hemophagocytic syndrome was established according to the diagnostic criteria laid down by the Histiocyte Society. We discuss the diagnosis and management of this complex case and try to generate awareness about dengue induced HLH as one of the possible causes for severe manifestations of this infection.

A prospective study of ventilator-associated tracheobronchitis: Incidence and etiology in intensive care unit of a tertiary care hospital.

CONTEXT: Ventilator-associated tracheobronchitis (VAT) is an infective complication of mechanical ventilation and is a part of the spectrum of ventilator-associated respiratory infections. In the Intensive Care Units (ICUs), VAT is a relatively common problem but in comparison to ventilator-associated pneumonia (VAP), much less data are available on VAT and its management. MATERIALS AND METHODS: Patients ventilated for more than 48 hours were screened daily for the development of VAT. Patients were followed up daily until they were extubated, died or discharged from the hospital. The patient demographics, underlying condition, causative organism and resistance patterns were observed. RESULTS: 13.2% of patients developed VAT. The majority patients who developed VAT had underlying neurological problems. The mean time to develop VAT from the time of mechanical ventilation was 7.3 days and from time of ICU admission was 10 days, respectively. Multidrug-resistant (MDR) Acinetobacter sp. and Pseudomonas aeruginosa were the most frequently isolated organisms. CONCLUSIONS: VAT is a common healthcare-associated infection caused mostly by MDR Gram-negative bacteria. Monitoring and active surveillance are required to detect VAT at the earliest to institute appropriate isolation measures and therapy.

Invasive infection due to Moraxella lacunata, an unusual pathogen.

Moraxella lacunata, a commensal bacterium, is associated with serious invasive disease. We describe a patient with diabetic nephropathy who developed septicemia with metastatic abscesses in the liver and spleen due to Moraxella lacunata. The patient also had multiple ring enhancing lesions in both the cerebral hemispheres, possibly due to the same organism.

An Unusual Case of Septic Arthritis of the Hip.

Non-typhoidal Salmonella, particularly Salmonella enterica serovar typhimurium is food borne pathogen causing mild self-limiting diarrhoea in healthy adults. It can occasionally cause extraintestinal focal infection in susceptible patients. Salmonella, as the aetiological agent of osteomyelitis and septic arthritis is rare and has been mostly reported in patients with sickle cell disease or thalassaemia. We report a case of septic arthritis by Salmonellatyphimurium in an immunocompromised patient who was successfully treated following timely isolation and identification of the aetiological agent.

Revisit of fluoroquinolone and azithromycin susceptibility breakpoints for Salmonella enterica serovar Typhi.

In recent years, increase in occurrence of fluoroquinolone (FQ)-resistant S almonella Typhi isolates has caused considerable inconvenience in selecting appropriate antimicrobials for treatment of typhoid. The World Health Organization (WHO) recommends azithromycin for the empirical treatment option of uncomplicated typhoid. The CLSI updated the breakpoints of disc diffusion (DD) and MIC results of FQs and azithromycin for Salmonella Typhi in 2015, but DD breakpoints of ofloxacin and levofloxacin were not included. In this study, the inhibition zone diameters and MICs of nalidixic acid, ciprofloxacin, ofloxacin, levofloxacin and azithromycin were determined in Salmonella Typhi Kolkata isolates (n =146) over a 16-year period (1998 to 2013) and the data were compared with the available CLSI breakpoints. Very major error and major error (ME) of FQs were not observed in the study isolates, but the minor error of ciprofloxacin (15.8 %) and ME of azithromycin (3.5 %) exceeded the acceptable limit. A positive correlation between MICs of FQ and mutations in the quinolone-resistance-determining region (QRDR) showed the reliability of MIC results to determine FQ susceptibility of Salmonella Typhi (n =74). Isolates showing decreased ciprofloxacin susceptibility (MIC 0.125-0.5 µg ml-1) were likely to have at least one mutation in the QRDR region. The results on DD breakpoints of ofloxacin (resistant, ≤15 mm; intermediate, 16-24 mm, and susceptible, ≥25 mm) and levofloxacin (resistant, ≤18 mm; intermediate, 19-27 mm, and susceptible, ≥28 mm) corroborated those of earlier studies. In view of the emerging FQ- and azithromycin-resistant Salmonella Typhi isolates, DD and MIC breakpoints of those antimicrobials should be revisited routinely.