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Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM Objective The study aims to generate preliminary data about post-COVID pulmonary fungal infections in the Himalayas and analyze patients’ micro-radio-clinical profiles and outcomes. Methodology We conducted a retrospective study at a tertiary care teaching hospital in the Himalayas to generate preliminary post-COVID pulmonary fungal infection data. Sputum, Endotracheal Tube (ET), and Bronchoalveolar lavage (BAL) samples of patients sent to the Mycology laboratory were subjected to KOH mount and aerobic inoculation on Sabouraud dextrose agar plates at 37°C. The patients’ symptoms, diagnosis, clinical-radiological profile, and outcome were collected from the hospital database. Results Among n = 16 cases of post-COVID pulmonary fungal infections aged 53 +/- 13.38 years, n = 7 (43.75%) had Pulmonary Aspergillosis (n = 5 A. fumigatus, n = 1 A. flavus, n = 1 A. niger), n = 5 (31.25%) had Pulmonary Mucormycosis (Rhizopus arrhizus), and n = 4 (25%) had mixed infection. In 2 of 4 mixed infection patients, R. arrhizus was identified on KOH microscopy and A. fumigatus on SDA Agar. Both A. fumigatus and R. arrhizus were identified on KOH Microscopy of the third patient, while only A. fumigatus was cultivated on his SDA Agar. Aspergillus flavus and R. arrhizus were isolated simultaneously from the sample of the last patient, but only R. arrhizus was identified on KOH Microscopy. Clinical symptoms were similar among Pulmonary Aspergillosis and Mucormycosis patients, but hemoptysis was reported only among Pulmonary Aspergillosis patients. Pre-existing co-morbid end-organ damage, AKI, CKD, CLD, COPD, and CAD was more common among Pulmonary Mucormycosis patients and rare among Pulmonary Aspergillosis patients. Treatment requirements and clinical outcomes of patients infected with either mold were similar. The clinical profile of mixed infection patients was notably different from the others. All the patients were males, none complained of chest pain or expectoration, and none had a history of PTB, AKI, CKD, CLD, COPD, or CAD. Only 2 (50%) mixed infection patients needed supplemental high flow oxygen, unlike all (100%) patients diagnosed with single mold infection. None of the mixed infection patients required steroids. Moreover, none of the mixed infection patients died, unlike 60% mortality in cases of single-species infections. On radiological investigation, n = 6 had typical thick-walled cavitary lesions with air-fluid levels and multiple centrilobular nodules giving a tree in bud appearance, of which n = 4 had bilateral lung involvement, and n = 2 had only one lung involved. n = 1 patient had a well-circumscribed lung abscess. Conclusion COVID patients from the Himalayas had a higher prevalence of invasive pulmonary fungal infections, probably due to the dense surrounding vegetation. The immuno-compromised state following COVID-19 infection/treatment might be responsible for the progression of regular exposure to invasive pulmonary infection.

Introduction: The overall changing epidemiology of fungal infections in the current scenario isbecause of an increase in immunocompromised population including cancer patients, Humanimmunodeficiency virus (HIV)-infected patients, transplant receipts, and prolonged hospitalizationwith overuse of antimicrobial agents. These infections are challenging to diagnose and subsequentlymanage as their clinical symptomatology often mimics other common diseases like tuberculosis.Rapid diagnosis is limited and culture is often delayed due to slow growth rates of the causativeagents. Objective: This is a retrospective study to know the spectrum and burden of mycoticinfections in a tertiary care hospital. Methods: All samples collected from clinically suspected casesof fungal infections were sent to the Microbiology department over one year. The common specimensreceived were respiratory samples, scrapings from cornea, skin, and nail. All samples were firstobserved under direct microscopy using Potassium hydroxide (KOH) examination for the presence offungal elements and Gram stain for yeasts. India Ink examination was performed for sterile fluids.Fungal culture was done on Sabouraud's dextrose agar. Result: A total of 900 samples from variousdepartments were included, KOH examination was positive for 380 samples (42%) and fungalgrowth was obtained in 144 samples (16%). Rare fungi like Trichosporon dohaense (blood culture),Cladophialophora bantiana (brain abscess), Scedosporium apiospermum and Candida auris (bloodculture) were also isolated. Conclusion: Similar studies are needed to estimate the actual burden ofthe fungal infections in tertiary care health facilities, to help decrease the morbidity and mortalityassociated with underdiagnosed mycotic infections.

Background: Microbial keratitis is one of the most significant causes of blindness in our country.Knowledge regarding the aetiology and antibiotic sensitivity pattern in a specific region is crucial forthe ideal management of these infections. Materials and Methods: A prospective study wherecorneal scrapings from 161 suspected cases of microbial keratitis were collected and were subjectedto direct microscopy by gram stain and bacterial culture and identification as well as antibioticsensitivity testing. Results: A total of 57 samples turned out to be positive out of the total 161 andwere shared equally by both gram-positive and gram-negative bacteria. All these culture-positivebacteria were also identified by MALDI TOF-MS and were speciated. Few rare organisms which couldnot be identified by conventional means were also recovered using the same. Most of the gram-positive isolates showed good sensitivity to vancomycin and ciprofloxacin whereas Pseudomonas sppwas found to be resistant to the aminoglycosides. Conclusion: Right knowledge about the localprofile of bacterial causes of keratitis along with its antibiotic resistance pattern will help theclinicians immensely and help them to initiate the correct empirical therapy bases on the smearresults without wasting crucial time.

Background and Purpose: Herein, we describe a rare case of fatal cerebral phaeohyphomycosis by Cladophialophora bantiana in an immunocompetent individual without any underlying risk factors.Case report: A 55-year-old female presented with a short history of fever for 1 month, as well as headache, sudden onset of right-sided upper and lower limb weakness, and loss of speech for 10 days. Contrast-enhanced magnetic resonance imaging scan revealed large, peripherally enhancing, well-defined, cystic, space-occupying, axial lesion in the left parietal lobe with a mass effect. The patient was subjected to craniotomy, and the drained pus revealed pigmented septate hyphae in potassium hydroxide mount examination, which was identified as Cladophialophora bantiana. The patient was started on amphotericin B and voriconazole. However, she developed acute respiratory infection leading to multi-organ failure and death on day 27 post-operation. Conclusion: In the absence of comparative trials owing to the rarity of the disease, the radical resection of lesion, drainage of pus, and prolonged targeted antifungal therapy with close postoperative radiological surveillance are the therapeutic measures of choice for patients with brain abscess caused by phaeoid fungi.

Perinephric and renal abscess are very rare entities with insidious presentation and pose a great diagnostic challenge, common etiology being bacterial. Only less than 30 cases of fungal etiology have been reported in literature. Herein, we report first case of fungal perinephric abscess caused by Candida tropicalis in a young diabetic female who presented with right flank pain and vomiting. Diagnosis made by CT imaging and culture of USG guided aspiration of perinephric abscess revealed growth of Candida tropicalis. Candida tropicalis perinephric abscess requires a very high index of suspicion for diagnosis. Associated high morbidity and mortality rates are likely due to misdiagnosis, therefore fungal perinephric and renal abscess must be kept as differential diagnosis in cases of fever with abdominal pain. Prompt diagnosis and early treatment are important for better outcome. Imaging and microbiological investigations are required for diagnosis, and drainage is indicated for successful therapy.

Background: Hepatitis B and C cause serious infectious disease of the liver which affects millions of people worldwide. More than 2 billion people living today have been infected with Hepatitis B virus (HBV) at some time in their lives and about 350 million people are carriers of the virus. Aim: To detect HBV and HCV infection and assess HBV immunity status amongst healthcare workers, medical students and nursing students at AIIMS Rishikesh. Methods: 135 participants (Medical faculty, Nursing Faculty, Resident Doctors, laboratory technicians, laboratory attendants, medical and nursing students) were enrolled for the study. Serological tests HBsAg, anti-hepatitis C virus antibodies (anti-HCV), anti-HBs antibodies, were performed. None were positive for HBsAg 1.5% were positive for Anti HCV antibodies and 61.75% had protective levels of antiHBs. Conclusion: HBV vaccination and screening for HBV, HCV, and anti-HBs of all HCW and students at time of entry should be done. Yearly screening thereafter should be done to check as follow up. Regular training on HBV vaccination and safe practices should be emphasized at all levels of HCW.

Cryptococcosis is a fungal disease with worldwide distribution and wide array of clinical manifestations, caused by encapsulated basidiomycetous yeasts called Cryptococcus spp. It has traditionally been considered an opportunistic infection known to occur in immunocompromised hosts, particularly those who are infected with human immunodeficiency virus. However, this infection has also been reported in phenotypically 'normal' or otherwise clinically non-immunocompromised patients. The seemingly mysterious nature of this potentially fatal illness has always kept clinicians and diagnosticians in a dilemma. This case series reiterates this perspective.

Background: Infectious Keratitis (Ik) Is A Potential Vision-Threatening Ocular Infection Caused By A Variety Of Microorganisms. Aim: To Explore Risk Factors And Etiological Agents Associated With Ik. Design: A Prospective Cross-Sectional Study In Which Corneal Scrapings From 120 Suspected Cases Were Evaluated At A Tertiary Health Care Institute From January To December 2019. Methods: Scrapings Were Subjected To Direct Microscopy, Culture, And Identification By Both Conventional Methods And Maldi-Tof-Ms. The Patient's Demographic Data And Predisposing Factors, If Any Were Recorded. Results: The Mean Age Of Patients Was 48.9 Years And Predisposing Factors Were Documented In 46% (55/120) Of Cases. Overall, Infective Etiology Could Be Established In 51% (N = 61/120) Of Cases. Fungal Growth In 26% (N = 31/120) Of Cases And Bacterial Growth In 22% (N = 27/120) Of Cases Was Obtained. Growth Of More Than One Species Of Fungi Or Growth Of Bacteria Along With Fungus Were Observed In 2% (N = 3/120) Of Cases. Of All The Fungal Isolates Obtained (N = 34), The Most Common Isolate Was Fusarium (18/34) Followed By Aspergillus (8/34), Curvularia (4/34), Pseudallescheria Boydii (3/34), And Geotrichum (1/34). Among The Gram-Positive Bacterial Isolates (N = 16), Staphyloccus Species (15/16) Were Isolated In Maximum Number Followed By Streptococcus Pneumoniae (1/16). Among The Gram-Negative Isolates (N = 13), Pseudomonas Species (8/13) Were Isolated In Maximum Number Of Cases, Followed By Acinetobacter (3/13), Klebsiella Pneumoniae (1/13), And Escherichia Coli (1/13). Conclusion: For Initiating Appropriate Empirical Therapy, The Knowledge Of The Epidemiological Pattern Of Infectious Keratitis Of A Particular Geographical Region Is Crucial.

Introduction Invasive fungal infections have always been a major cause of mortality and morbidity and are especially prevalent in the immunosuppressed groups of patients. Members of the Mucoracea family have an increasing incidence and prevalence. It has always been difficult to diagnose this condition due to various reasons. Materials and Methods This was an observational study carried out jointly by the Department of Microbiology and the Department of Pathology for a duration of one year. All patients who presented in various clinical departments with a high index of clinical suspicion for mucormycosis were included in this study. A total of 186 samples were received from suspected cases of mucormycosis and were all subjected to direct microscopy by potassium hydroxide (KOH), fungal culture, and histopathological examination. Results Mucormycosis was documented in 33 out of 186 cases on direct microscopy, whereas 21 were positive on fungal culture. Histopathological positivity was reduced with only 11 cases showing aseptate hyphae suggestive of mucormycosis. Conclusion As these organisms generally do not grow well on routine culture media and with the histopathological results also being not suggestive clearly of mucormycosis, direct microscopy thus becomes more important and essential in the rapid diagnosis of this deadly condition.

Background and Purpose: We report a case of disseminated cryptococcosis in a treatment-naïve patient, incidentally diagnosed with hepatitis C virus (HCV) infection and renal parenchymal disease. The patient succumbed to death given the very late diagnosis of the disease. Case report: A 54-year-old male presented with the chief complaints of abdominal pain, chest pain, and phlegmy cough for a month. There was a past history of decreased urine output, lower limb swelling, and fever lasting for 15-20 days. After a general physical examination, the differential diagnosis of hepatitis C-related liver disease with hepatic encephalopathy, disseminated tuberculosis, and septic shock was made. Radiological examination revealed renal parenchymal disease on ultrasound abdomen and opacity with reticulonodular opacity in the bilateral lung zones. In laboratory investigations, serum reactive sample was tested for anti-HCV antibodies. In addition, Cryptococcus var grubii was identified in blood culture using the matrix-assisted laser desorption ionization-time of flight mass spectrometry (Bruker Daltonics, Germany). The patient succumbed to death before the initiation of any specific antifungal therapy. Conclusion: Cryptococcosis-HCV co-infection is a fatal condition with a fulminant course that might be difficult to treat.

Background and Purpose: More than 300 Fusarium species are grouped into approximately 23 species complexes out of which around 70 are involved in human infections. The nomenclature of these species has undergone considerable changes in recent years. These species cause localized infections in individuals while inducing systemic infections mainly in immunocompromised patients. The present study was conducted to identify Fusarium species in clinical isolates by molecular methods and determine their in vitro minimum inhibitory concentration (MIC) patterns to address the lack of data in this domain in Northern India. Materials and Methods: For the purpose of the study, Fusarium isolates obtained from various clinical samples were sent to the Westerdijk Fungal Biodiversity Institute, Utrecht, the Netherlands, for molecular identification. The MIC testing was performed using the microbroth dilution method as per the Clinical and Laboratory Standards Institute reference method (M38-A2). Results: Fusarium was isolated from 33 patients (i.e., 1, 1, 2, 14, and 15 cases with endophthalmitis, sinusitis, pulmonary involvement, onychomycosis, and keratitis, respectively). These 33 isolates belonged to three species complexes, namely F. solani species complex (FSSC; n=13), F. fujikuroi species complex (FFSC; n=13), and F. incarnatum equiseti species complex (FIESC; n=7). The species identified within FSSC, FFSC, and FIESC included F. keratoplasticum (n=6)/F. falciforme (n=6)/F. solani (n=1), F. proliferatum (n=7)/F. sacchari (n=5)/F. anthophilum (n=1), and F. incarnatum SC species (n=6)/F. equiseti SC species (n=1), respectively. The MIC results showed that all isolates had a lower MIC against amphotericin B than against the other antifungal agents. Conclusion: Timely diagnosis and appropriate treatment will facilitate the improvement of patient outcomes.

Background: Candida species are generally identified by conventional methods such as germ tube or morphological appearance on cornmeal agar (CMA), biochemical methods using API kits, and molecular biological techniques. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) has revolutionized the identification of fungi reducing the turnaround time of days to minutes. Purpose: To compare the performance of MALDI-TOF MS and conventional methods in the identification of clinically relevant yeasts. Materials and methods: In this study, Candida identifications on CMA are compared with the results obtained on MALDI-TOF MS (Bruker Daltonics, Bremen, Germany). Discrepant results were confirmed by sequencing internal transcribed spacer (ITS) regions of rDNA. Results: A total of 114 clinical Candida species isolated from blood cultures were isolated and identified with conventional methods as well as with the MALDI-TOF-MS system. The agreement between the two test results were analyzed using Inter-rater reliability analysis (Cohen's Kappa) in SPSS Software Version 24 (IBM Corp., Armonk, NY). Overall, there was substantial agreement (Cohen's kappa=0.763) between the two methods. A value between 0.61 and 0.80 is classified under substantial. The most frequently isolated bloodstream Candida species included Candida albicans, C. tropicalis, C. parapsilosis, C.lusitaniae, C. glabrata which were accurately identified by MALDI-TOF-MS. When compared with conventional identification methods, MALDI-TOF-MS results are more reliable and rapid for Candida identification.

Fungal infections are life threatening especially in presence of immunosuppression or uncontrolled diabetes mellitus mainly due to their invasive potential. Mucormycosis of the oculo-rhino-cerebral region is an opportunistic, aggressive, fatal and rapidly spreading infection caused by organisms belonging to Mucorales order and class Zygomycetes. The organisms associated are ubiquitous. Aspergillosis is a common clinical condition caused by the Aspergillus species, most often by Aspergillus fumigatus (A. fumigatus). Both fungi have a predilection for the immunosuppressive conditions, with uncontrolled diabetes and malignancy being the most common among them. Mucormycosis is caused by environmental spores which get access into the body through the lungs and cause various systemic manifestations like rhino-cerebral mucormycosis. Here, a case series of such concomitant infections of Aspergillus and Mucor spp from Rishikesh, Uttarakhand, India is reported.

Background Cerebral aspergillosis usually affects immunocompromised hosts and may rarely occur in immunocompetent individuals. Due to its angio-invasive nature, Aspergillus may cause various vascular complications, particularly mycotic aneurysms and infarcts. Case presentation A 22-year-old immunocompetent male with diagnosed case of sino-cerebral aspergillosis was taking voriconazole for two months. His headache worsened and repeat imaging showed an increase in the size of the lesion. The patient was managed with right frontal craniotomy and surgical debridement, and voriconazole was continued. After ten days of uneventful post-operative course, the patient developed left-sided hemispheric infarct. The patient is doing well at nine months' follow-up, and he is off voriconazole for three months after the follow-up imaging showed complete resolution of disease. Conclusion Treatment of choice for cerebral aspergillosis is voriconazole. Surgical debridement may be a useful adjunct in patients not responding to voriconazole alone.

Although Candida albicans remains the predominant species causing Blood Stream Infection (BSI), recent studies have demonstrated an emergence of Non-albicans Candida spp (NAC), such as C. glabrata, C. papapsilosis and C. krusei. Candida krusei are yeast-like microorganisms which may colonise the human skin, respiratory or gastrointestinal tract but can cause lethal infections especially in immunocompromised patients. Hereby, author’s report a case of 10-year-old male with T-cell Acute Lymphocytic Leukaemia (ALL) who reported with fever and non-productive cough caused by Candida krusei. Direct gram stain from blood culture and germ tube test was performed. Further, the isolate was initially misidentified by Matrix Assisted Laser Desorption/Ionisation Time of Flight Mass Spectrometry (MALDI-TOF MS) as Trichosporon ovoides which was later identified by molecular sequencing as Candida krusei.

Nocardiosis is a rare bacterial infection that may lead to a severe disease. These infections are rare among normal population and are showing an increasing trend worldwide attributable to the increase in the immunosuppressed population. Most of these patients present with nonspecific clinical features such as fever, productive cough, and exertional dyspnoea as seen in our series of patients which makes it difficult to be diagnosed. Pulmonary nocardiosis is rarely clinically suspected and often diagnosed very late in the course of disease resulting in high mortality. A similar observation was made in one of our cases where the patient was being treated on the lines of pneumonia, and in the end she was diagnosed with pulmonary nocardiosis. In view of the limited literature available, we report here a case series of pulmonary nocardiosis in immunosuppressed patients diagnosed incidentally by fungal KOH mount. The most common conditions causing immunosuppression were type II DM, COPD, and pulmonary tuberculosis.

Background and Purpose: More than 300 Fusarium species are grouped into approximately 23 species complexes out of which around 70 are involved in human infections. The nomenclature of these species has undergone considerable changes in recent years. These species cause localized infections in individuals while inducing systemic infections mainly in immunocompromised patients. The present study was conducted to identify Fusarium species in clinical isolates by molecular methods and determine their in vitro minimum inhibitory concentration (MIC) patterns to address the lack of data in this domain in Northern India. Materials and Methods: For the purpose of the study, Fusarium isolates obtained from various clinical samples were sent to the Westerdijk Fungal Biodiversity Institute, Utrecht, the Netherlands, for molecular identification. The MIC testing was performed using the microbroth dilution method as per the Clinical and Laboratory Standards Institute reference method (M38-A2). Results: Fusarium was isolated from 33 patients (i.e., 1, 1, 2, 14, and 15 cases with endophthalmitis, sinusitis, pulmonary involvement, onychomycosis, and keratitis, respectively). These 33 isolates belonged to three species complexes, namely F. solani species complex (FSSC; n=13), F. fujikuroi species complex (FFSC; n=13), and F. incarnatum equiseti species complex (FIESC; n=7). The species identified within FSSC, FFSC, and FIESC included F. keratoplasticum (n=6)/F. falciforme (n=6)/F. solani (n=1), F. proliferatum (n=7)/F. sacchari (n=5)/F. anthophilum (n=1), and F. incarnatum SC species (n=6)/F. equiseti SC species (n=1), respectively. The MIC results showed that all isolates had a lower MIC against amphotericin B than against the other antifungal agents.

The two most common filamentous fungi causing mycotic keratitis are Aspergillus and Fusarium spp. Around 70 Fusarium spp. are involved in causing human infections. In this study, four cases of keratitis in sugarcane farmers in India are being reported, caused by the sugar cane pathogen Fusarium sacchari, a species of the Fusarium fujikuroi species complex. Fusarial keratitis was established by potassium hydroxide/Calcofluor white wet mounts and fungal culture of corneal scrapings on conventional media. Final identification was done by genetic sequencing at CBS-KNAW, Utrecht, The Netherlands. The antifungal susceptibility testing was done using broth microdilution method as per CLSI document M38-A2. Four cases of F. sacchari keratitis were identified. Three of them had trauma with sugarcane leaves, whereas one sugarcane farmer reported trauma by vegetative matter. The morphological similarities among various Fusarium species warrant use of molecular methods for identification of cryptic species. A wide distribution of sugarcane farming could be the possible explanation for emergence of F. sacchari keratitis in India.

Background: Cryptococcal meningitis is a fatal opportunistic neuroinfection and an AIDS defining illness. It can also occur in non-HIV patients who are immunodefecient due to chronic glucocorticoid use, organ transplantation, malignancy and sarcodiosis. Materials and Methods: A cross-sectional study was conducted in a tertiary care hospital from July to December 2018. CSF samples of 364 patients were received by Microbiology laboratory during this period for the purpose of aerobic bacterial, fungal and TB culture, respectively. All samples were subjected to examination by direct wet mount, Gram stain and India ink preparation. Ziehl Neelsen staining, solid culture for Mycobacterium tuberculosis on Lowenstein Jensen medium and Gene Xpert was also performed on all CSF samples. These samples were further subjected to fungal culture on Sabouraud's dextrose agar. Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) was used for identifying all bacterial (except M. tuberculosis) and fungal isolates. Results: Out of 364 CSF samples received, 288 were sterile after 48 hours of aerobic incubation. Bacterial isolates, M. tuberculosis and Cryptococcus spp. were obtained in culture from 51, 21 and 4 samples, respectively. The prevalence of cryptococcal meningitis in our study was 1.09% (4/364). Cryptococcus neoformans var grubii was the most common isolate (2/4; 50%) followed by Cryptococcus neoformans var neoformans (1/4; 25%) and Cryptococcus neoformans var gattii (1/4; 25%), respectively. Conclusion: Cryptococcal meningitis is a rapidly fatal condition which requires a high index of suspicion and calls for a collective effort from family physicians and diagnosticians alike. This disease is under-reported from Uttarakhand and therefore calls for further research from this region.

The presence of Candida in urine presents a therapeutic challenge for the physician as it is often asymptomatic, and management guidelines have not been clearly laid down on this issue. The presence of Candida in urine may represent contamination of clinical sample, actual colonization of the lower urinary tract or may be a true indicator of invasive infection of lower and/or upper urinary tract. In a clinical setting like the ICU, multiple risk factors for Candida colonization may be present in the same patient, thereby increasing the chances of candiduria, manifold. In the present study on 80 patients in ICU, high rate of Candida colonization (57.5%) was found in urine samples of ICU patients with C. tropicalis (57.3%) being the predominant species. We also isolated 8 strains of Trichosporon species, all of these presented as a mixed infection along with Candida species. Among the various risk factors studied, urinary catheterization and previous antibiotic therapy were identified as statistically significant (P value

Salmonella are most commonly associated with gastroenteritis and enteric fever in humans. Occasionally, dissemination of bacilli throughout the body results in establishment of localized foci of persistent infection especially in patients with debilitating diseases and immunosuppressive states. Infection at various aberrant sites due to Salmonella has been reported relatively seldom. It has perfected the art of intracellular survival in niches from where they can cause myriad of effects. Six cases with Salmonella infection at unusual sites without any preexisting or underlying disease diagnosed over a period of two years are presented here. Salmonella etiology was not suspected in these patients and the diagnosis was made microbiologically only after culture isolation.

Dear Editor Now-a-days, anaerobic infections are showing evidence of increasing virulence, rising incidence, unresponsiveness to metronidazole therapy and worse outcomes. Some of these infections are serious and have high mortality rate and can no longer be overlooked as in the past and need to be properly identified [1]. Anaerobic microbiology has fallen out of the spotlight of infectious disease, due to extraordinary efforts required to recognize these infections as also the availability of generally effective antimicrobials against these organisms [2]. Anaerobic culturing can be made cost efficient by strict adherences to several principles, including the selective culturing of only appropriate general specimens that are uncontaminated by normal flora; Rapid transport of specimens and use of appropriate transport system; use of a system of rejection when inappropriate and multiple samples have been received [3]. This was a prospective study conducted at a Department of Microbiology, Government Medical College and Hospital Chandigarh, India over a period of three years from June 2009 to June 2012. The study comprised analysis of 100 pus samples from patients suspected to have anaerobic infections from Diabetic foot ulcers, Head and neck, Breast, Brain, Psoas abscesses, Peritoneal aspirations, Vaginal swabs received from different Departments of hospital. These included 5 samples in 2009, 14 samples in 2010, 40 samples in 2011 and 36 samples up to June 2012. The specimens were collected by aspiration in a sterile syringe or in the form of swab which were further transferred to transport media, Robertson cooked meat media (RCM). These samples were sent to the laboratory and processed immediately within 1-2 h of collection. Cultures were put up on brain heart infusion agar supplemented with haemin and vitamin K, L-cysteine, yeast extract with preliminary disks like metronidazole (5 μg), vancomycin (5 μg) and colistin (10 μg) sodiumpolyanethol sulphonate (SPS) discs for anaerobic incubation, Blood agar and MacConkey agar were put up for aerobic incubation. Incubation of brain heart infusion agar was done at 37°C for 48-72 h in anaerobic jar. Anaerobiasis was created by automated anaerobic system (Anoxomat). Aerobic plates were examined after 24 h. Anaerobic plates were examined after 48-72 h and observed for any growth. Colony characteristics were noted and staining was done and those organisms which were sensitive to metronidazole were considered as anaerobes. However some colonies also showed resistance to metronidazole. These individual colonies were inspected and aerotolerance test was done for each of them. Those organisms which failed to grow aerobically after 24 h on blood agar are considered as anaerobes. Pure culture isolates were identified by standard biochemical methods [3]. Antimicrobial susceptibility testing was done with various commonly used antimicrobial agents that are recommended by CLSI for anaerobes by the disc diffusion method [4]. Approval for study has been taken from ethical committee of the department. Isolation rate of anaerobic bacteria was 19%. The predominant anaerobic bacteria were Peptostreptococcus species. (11%), followed by Eubacterium species (3%), Clostridium species (2%), Veillonella species (1%), Bacteroides fragilis (1%), Prevotella species (1%). Anaerobic isolates are shown in [Table/Fig-1]. [Table/Fig-1]: Anaerobic isolates from samples The susceptibility of these bacteria was as follows: all the isolates were found to be sensitive to penicillin, clindamycin, imipenem, amoxycillin – sulbactam, piperacillin – tazobactum. Sensitivity to metronidazole was 84% (n-16/19). All the three isolates resistant to metronidazole were found to be Eubacterium species. The importance of isolation and characterization of anaerobes has long been overlooked. Certain reasons being the difficulty in isolation, time required and low levels of drug resistance. In our own institution we initially found great reluctance in sending samples but over a span of 3 y the number of samples increased from a mere 5 to a maximum of 40 in 2011 and until June 2012 already 36 samples had been received. This has given a new impetus to our anaerobe section and has been indeed encouraging. Additionally, good isolation results have been achieved. The prevalence of anaerobes in this study was 19%. The most commonly isolated anaerobe was Peptostreptococcus 11% followed by Eubacterium species 3%, Clostridium species 2% and Veillonella species, Bacteroides species, Prevotella species making up 1% each. The studies of Eslami et al., Colaycoet et al., reported Peptostreptococcus as the most frequently isolated anaerobe [5,6]. Most of the anaerobes were 100% sensitive to majority of the drugs; however, sensitivity to metronidazole was 84%. The resistance to metronidazole was shown by all the three Eubacterium species which are usually resistant to this drug despite the generally excellent activity of this drug [3]. The most frequently isolated antibiotic resistant anaerobe is Bacteroides fragilis due to beta-lactamase production [7]. The another study showed, metronidazole had the highest rate of resistance among anaerobes at a high 48% [8].This poses a difficulty since metronidazole is a frequent choice for empirical anaerobic coverage over the other antibiotics. Another cause of concern is the high resistance rate to clindamycin. But in our present study, no resistance to clindamycin was noted by any of the isolates. There is necessity for precautions to be taken for proper collection and transport of specimens where ever anaerobic bacteria are suspected to be the cause. For this trained paramedical staff is required. Therefore, the greatest challenge is to sensitize clinicians to send samples followed by the second hurdle of isolation, characterization and identification and routine sensitivity testing. Hence, clinicians and microbiologists working in tandem can go a long way in decreasing mortality due to anaerobic infections.

The incidence of multidrug resistant enteric fever is increasing alarmingly. This study was planned to determine the rate of isolation of Salmonella spp. and to compare the isolates for their epidemiological parameters and antimicrobial susceptibility patterns at our center.The study was conducted over a span of three years with a total of 8142, 8134, and 8114 blood culture samples processed for the years 2008, 2009, and 2010 respectively. The minimum inhibitory concentration (MIC) for ciprofloxacin and chloramphenicol was determined using an agar dilution method. The MIC for ciprofloxacin was also confirmed by Epsilon-test (E -test) strips.Of the total 302 Salmonella spp. isolated, 257 were Salmonella enterica serotype Typhi (85.1%) and 45 (14.9%) were S. enterica serotype Paratyphi A. The majority of the isolates recovered were from the pediatric age group (54.6%) and males (60.6%). Complete susceptibility was observed to chloramphenicol, cefotaxime, ceftriaxone, and azithromycin over the last two years (2009 and 2010), with an increase in resistance to nalidixic acid (100%) and ciprofloxacin (13.6%).In our study, we found augmentation of resistance to nalidixic acid and fluoroquinolones and complete sensitivity to ceftriaxone along with reemergence of chloramphenicol sensitivity for Salmonella isolates. This report emphasises the necessity of continuous surveillance of antibiograms of enteric fever isolates in an area.

Author(s): Dhingra, M; Kaistha, N; Bansal, N; Solanki, LS; Chander, J; Thami, GP; Sande, WW van de