Cladosporium halotolerans: Exploring an Unheeded Human Pathogen.

BACKGROUND: Cladosporium halotolerans is a saprobic fungus, rarely implicated in human infections. The identification is challenging due to non-specific phenotypic features. OBJECTIVE: To decipher clinical spectrum, microbiological and susceptibility profile of clinical and environmental isolates of Cladosporium halotolerans. METHOD: All the isolates identified as Cladosporium halotolerans deposited in National Culture Collection for Pathogenic Fungi (NCCPF), Postgraduate Institute of Medical Education and Research, Chandigarh, India were revived. Phenotypic and molecular characterization targeting internal transcribed spacer (ITS) region of ribosomal DNA, large subunit of ribosomal DNA (LSU; NL1 and NL4), actin (ACT) and beta-tubulin (TUB) was done. Scanning electron microscopy (SEM) was performed to determine any phenotypic variations. Antifungal susceptibility testing (AFST) was carried out for eight antifungal agents as per CLSI M38 Ed3 guidelines. We also performed systematic literature review of all the cases of Cladosporium halotolerans reported till date. RESULTS: A total of four isolates (clinical, n = 3; soil, n = 1) identified as Cladosporium halotolerans were included in the study. The clinical sites were skin, maxillary tissue and nail. All patients were apparently immunocompetent, and history of trauma was recorded in one patient. All patients improved on antifungal therapy. The cultures revealed growth of black mycelial fungus and microscopic examination demonstrated dematiaceous septate hyphae with erect conidiophores and conidia in branched acropetal chains. Based on molecular methods, all the four isolates were identified as C. halotolerans. SEM revealed no variation in length and width of the conidia, conidiophores, ramoconidium and hyphae among the isolates. All molecular targets, such as ITS region, LSU (partially sequenced), ACT and TUB were able to differentiate the isolates. Minimum inhibitory concentrations for antifungals were: triazoles (0.12-2 µg/ml), amphotericin B (4 µg/ml) and echinocandins (2-8 µg/ml). CONCLUSION: We report role of the rarely isolated dematiaceous fungus, C. halotolerans, in causing human infections. The study emphasizes the role of molecular methods in precisely identifying these species. Triazoles are more active against these black fungi compared to polyenes or echinocandins.

Changing Trends in Antimicrobial Susceptibility Patterns of Bloodstream Infection (BSI) in Secondary Care Hospitals of India.

Introduction Bloodstream infection (BSI) and subsequent sepsis are life-threatening medical conditions. The onset of antimicrobial resistance and subsequent multi-drug resistant organisms (MDRO) significantly increase healthcare-associated expenditure with adverse clinical outcomes. The present study was undertaken to identify the trends of BSI in community settings in secondary care hospitals (smaller private hospitals and district hospitals) in the state of Madhya Pradesh in Central India with the support of the Indian Council of Medical Research (ICMR) and National Health Mission, Madhya Pradesh. Methodology The present study was a prospective, longitudinal observational chart review type of study. The study was carried out at 10 secondary care hospitals (eight smaller private hospitals and two government district hospitals) nominated by the State Government as part of the ICMR Antimicrobial Resistance Surveillance and Research Network (AMRSN). The hospitals were nominated depending on the availability of a microbiology laboratory and a full-time microbiologist. Result A total of 6202 blood samples were received from patients with suspected BSI, out of which 693 samples were positive for aerobic culture. Among these, 621 (89.6%) showed bacterial growth and 72 (10.3%) grew Candida species (spp). Out of the 621 bacterial growth samples, Gram-negative bacteria were 406 (65.3%) and Gram-positive bacteria were 215 (34.6%). Among the Gram-negative isolates (406), the predominant isolate was Escherichia coli (115; 28.3%) followed by Klebsiella pneumoniae (109; 26.8%), Pseudomonas aeruginosa (61; 15%), Salmonella spp. (52; 12.8%), Acinetobacter spp. (47; 11.6%) and the other Enterobacter spp. (22; 5.4%). Among the Gram-positive isolates (215), the predominant isolate was Staphylococcus aureus (178; 82.8%) followed by Enterococcus spp. (37; 17.2%). Among the Escherichia coli, third-generation cephalosporin resistance was identified in 77.6%, piperacillin-tazobactam resistance in 45.2%, carbapenem resistance in 23.5% and colistin resistance in 16.5% of cases. Among the Klebsiella pneumoniae, third-generation cephalosporin resistance was identified in 80.7%, piperacillin-tazobactam resistance in 72.8%, carbapenem resistance in 63.3% and colistin resistance in 14% of cases. Among the Pseudomonas aeruginosa, ceftazidime resistance was identified in 61.2%, piperacillin-tazobactam resistance in 55%, carbapenem resistance in 32.8%, and colistin resistance in 38.3% of cases. Among the Acinetobacter spp., piperacillin-tazobactam resistance was identified in 72.7%, carbapenem resistance in 72.3%, and colistin resistance in 9.3% cases. While analyzing the antibiogram for Staphylococcus aureus isolates, methicillin resistance (MRSA) was seen in 70.3% of cases, followed by vancomycin resistance (VRSA) in 8% of cases and linezolid resistance in 8.1%. Among the Enterococcus spp. isolates, linezolid resistance was found in 13.5%, vancomycin resistance (VRE) in 21.6%, and teicoplanin resistance in 29.7% of cases. Conclusion In conclusion, the first-ever study to identify the risk of high-end antibiotics causing significant drug resistance in secondary and tertiary care settings has highlighted the urgent need for more randomized control studies and proactive measures from healthcare authorities and serves as a beacon for future research efforts and underscores the importance of implementing antibiograms to combat the growing threat of antibiotic resistance.

Foldscope: Diagnostic Accuracy and Feasibility of its Use in National Malaria Control Program.

Background: Malaria has been an important public health all over the globe. Although conventional light microscopy is the gold standard of diagnosis, light microscopes are heavy, fragile, costly, and electricity dependent. Rapid diagnostic tests (RDTs) have become more popular but perform badly in temperate climate. This is because the RDT kits require maintenance of cold chain for its optimal use. In this regard, there is a recent interest in handheld malaria microscopy at the point of care in the field setting. Foldscopes are cheap, handy, nonfragile, and use mobile camera for illumination. The purpose of the study was to find whether foldscope can be used in the national vector borne disease control program (NVBDCP) in India. Methods: Ten laboratory technicians were trained in identifying malaria parasites using foldscope and their mobiles. Later, they were provided with unassembled foldscope to document their test results for the preidentified malaria slides. The blood smears were stained as per the protocol of NVBDCP. The report of the index test (foldscope microscopy) was compared with the reference test (conventional microscopy). Results: The sensitivity and specificity of the index test was found to be 13.3% (6.257-26.18), specificity of 97.78% (88.43-99.61), positive predictive value 85.71% (48.69-97.43), and negative predictive value 53.01% (42.38-63.38). The devise failure rate and test failure rate were 20% and 11.7%. The kappa agreement between the index and reference microscopy was only 11% and the McNemar P < 0.01. Conclusion: The ×400 foldscope at its present magnification and illumination cannot be utilized in the field under NVBDCP.

Preliminary Experience in Post-COVID-19 Mycoses: A Pathologist's Perspective.

Background Coronavirus disease is caused by the severe acute respiratory syndrome coronavirus-19. Because of co-morbidities and indiscriminate use of steroids and antibiotics, the incidence of opportunistic fungal infections has increased in COVID-affected individuals. Aims and objectives The aim of the study is to analyze the various tissue reaction patterns of COVID-19-associated mucormycosis in the surgical debridement specimens using routine hematoxylin and eosin (H&E) stain and special stains like periodic acid-Schiff (PAS), Grocott-Gomori's methenamine silver (GMS), Masson trichrome (MT) and Prussian blue (PB), and to understand the pathogenesis of COVID-19 sequelae. Materials and methods This retrospective observational study was conducted after the approval from the Institute Human Ethical Committee (IHEC) on 45 tissue samples of COVID-associated mucormycosis using routine H&E and histochemical stains such as PAS, GMS, MT, and PB. Detailed demographic profiles, clinical information, radiological findings, and relevant microbiological data in available cases, like reports on potassium hydroxide (KOH) mount preparation, and fungal culture reports on Saboraud's Dextrose Agar (SDA) medium were collected. The different histomorphological tissue reaction patterns were observed and analyzed. Results All the surgical debridement specimens from post-COVID cases had histomorphology of mucormycosis displaying broad, aseptate, ribbon-like fungal hyphae with right-angle branching (45/45). Six of the 45 cases also reveal thin, narrow septate, acute angle branching hyphae, indicating co-existing Aspergillosis (6/45). The histological tissue reaction patterns observed were categorized as extensive tissue necrosis (100%), vascular proliferation (82%), angioinvasion (58%), giant cell reaction (53%), fibrin thrombi (47%), septic thrombi and angiodestruction (40%), fungal osteomyelitis (33%), necrotizing granulomas (31%). Conclusion This study infers that post-COVID-19 associated mucormycosis, alterations in the local tissue microenvironment are found to have a favorable effect on colonizing fungi and result in destructive tissue reactions such as angioinvasion, angiodestruction, necrosis, necrotizing granulomas, suppurative inflammation, and iron pigment deposition. The spectrum of morphological changes reflects the host's immune status.

Clinico-Epidemio-Microbiological Exploratory Review Among COVID-19 Patients with Secondary Infection in Central India.

Purpose: Secondary infections (SI) in COVID-19 have been documented from 3.6% to 72% in various studies with mortality ranging from 8.1% to 57.6%. There is a gap in knowledge for clinico-epidemio-microbilogical association among COVID-19 patients with concomitant SI. Patients and Methods: This is a retrospective chart review, in central India. The study was undertaken for hospitalized adult patients during 1st June 2020 to 30th November 2020, with laboratory proven COVID-19 infection and secondary infection. Results: Out of the total 2338 number of patients, only 265 (11.3%) patients were investigated for microbiological identification of SI. Male gender was predominant (76.8%) and the mean age was 53.7 ± 17.8 years. Only 3.5% (82/2338) of patients were having microbiologically confirmed (bacterial or fungal) SI. The overall mortality was 50.9% (54/82) with a differential mortality of 88.8% (48/54) in high-priority areas and 21.4% (6/28) in low-priority areas. Blood was the most commonly investigated sample (56%) followed by urine (20.7%) and respiratory secretion (15.8%). A. baumanii complex (20/82, 24.3%) was the most common bacteria isolated followed by K. pneumonia (12/82, 14.6%) and E. coli (11/82, 13.4%). Candida spp. (20/82, 24.3%) was the most common fungal pathogen isolated. Sixty percent (12/20) of Acinetobacter spp. were carbapenam-resistant and 70.3% of Enterobacterales were carbapenam-resistant. Fluconazole resistant Candid a spp. was isolated only in 10% (2/20) of cases. Diabetes was the most common co-morbidity 54.8% (45/82) followed by hypertension (41.4%) and chronic heart disease (13.4%). The negative predictors of secondary infections are urinary catheterization, placement of central line and mechanical ventilation (invasive and non-invasive). Conclusion: There is an urgent need of better anti-microbial stewardship practices in India (institutional and extra institutional) for curtailment of secondary infection rates particularly among COVID-19 patients.

Impact of an Anti-Microbial Stewardship Program on Targeted Antimicrobial Therapy in a Tertiary Care Health Care Institute in Central India.

Introduction Antimicrobial resistance (AMR) has become a global pandemic. In order to identify this menace, World Health Organisation (WHO) has developed the Global Action Plan on AMR (GAP AMR). Antimicrobial stewardship programs (AMSP) have been identified as a decisive tool for combating AMR. One of the most efficient measures of these programs has been the implementation of point prevalence surveys (PPS) of antibiotic usage and subsequent audit feedback. The present study was undertaken to identify the impact of AMSP on curtailing of empirical usage of antibiotics and the augmentation of targeted therapy.  Methods It is an observational, cross-sectional study comprising 1396 patients. The microbiology culture details and anti-microbial-sensitivity results were recorded. Antibiotic prescriptions were recorded in each patient during their hospital stay. Result Out of 1396 patients treated over four quarters (Q1-Q4), 711 (50.9%) patients were on antibiotics, and among them, only 415 patients were subjected to any microbiological cultures with an overall bacterial culture rate (BCR) of 58.3%, and 296 patients (41.6%) were treated with antibiotics empirically without sending any samples for bacterial culture. There was a statistically significant rise in BCR from 47.3% in the first quarter to 77.6% in the fourth quarter. Sending specimens for blood culture increased significantly from 29.2% in Q1 to 37.6% in Q4. After receiving culture reports, 72.3% of cases continued with the same antibiotic, the antibiotic was changed in 19.9% of cases, and the antibiotic was stopped in 7.8% of cases. Conclusion There was a strong positive impact of AMSP in curtailment of empirical usage of antibiotics and augmenting targeted therapy as evidenced by the significant rise in BCR over Q1-Q4 PPS as well as a significant rise in ordering for blood culture over the same time period.

A Pilot Study for the Evaluation of PCR as a Diagnostic Tool in Patients with Suspected Dermatophytoses.

CONTEXT: The conventional diagnostic tools for dermatophytoses suffer from several limitations including low sensitivity, specificity, and long turn-around-time. AIMS: The present study was, therefore, performed to evaluate the performance of a polymerase chain reaction (PCR)-based method for the diagnosis of this condition. SETTINGS AND DESIGN: The study was conducted in the Dermatology outpatient department of a tertiary care teaching hospital in central India over a period of 3 months from July to September 2015. MATERIALS AND METHODS: Forty participants, including 25 cases and 15 controls, were recruited in this observational study. Direct microscopy and fungal culture were performed from skin scrapings and nail clippings collected from the participants. PCR was also performed to amplify the chitin synthase 1 and internal transcribed spacer 2 genes from DNA samples extracted from the same clinical materials, using the method reported by Brillowska-Dabrowska et al. The diagnostic performance of fungal culture and PCR was compared using OpenEpi software. RESULTS: We observed a significant male predominance among patients with dermatophytoses. The sensitivity of fungal culture and dermatophyte PCR to diagnose dermatophytoses was 24% and 48%, respectively, whereas the specificity of the two assays was 100% and 93.3%, respectively. The likelihood ratio of a positive PCR assay was 7.2 and the negative likelihood ratio was 0.5. PCR assay also delivered a significant shortening of the time-to-diagnosis, with the mean turn-around-time being 8 hours and 14 days for PCR and culture, respectively. CONCLUSION: This study, thus, highlights the potential merits of the dermatophyte PCR assay in achieving a rapid diagnosis of dermatophytoses and underscores its utility as a complementary test to improve the sensitivity of the conventional diagnostic tools for this condition, as well as to reliably differentiate this condition from other similar skin conditions.

A rare case of urinary balantidiasis in an elderly renal failure patient.

Balantidium coli is the largest ciliated protozoa infecting humans by the feco-oral transmission from pigs. Large gut is the most common site of involvement. Symptomatology varies from asymptomatic carrier to invasive dysentery. Extra-intestinal infections can occur in liver, lung and urogenital tract. There are very few case reports of urinary balantidiasis. We present a case of urinary balantidiasis in an elderly farmer having diabetes and chronic kidney disease. This case is reported for its rarity and future references.

Severe Falciparum Malaria-Difference in Mortality among Male and Nonpregnant Females.

BACKGROUND: There are many studies regarding the gender difference in severe falciparum malaria. But most of these studies indeed refer to pregnant females only. There are scanty and conflicting reports regarding the mortality trend in nonpregnant females. MATERIALS AND METHODS: This is a case control study which is open, single centre, single blind, prospective and interventional study. SPSS 19 was used at the end of the study for all statistical analysis. Student's t-test was used for nominal variables and chi-squared test was used for categorical variables. RESULTS: The mean pre hospitalization illness period is significantly lower in females. Incidence of convulsion is significantly higher in females. The mean haemoglobin concentration in females is significantly lower. The mortality in females is significantly higher. Females over 40 years have higher mortality than females less than 40 years and also males over 40 years. Shock in female was associated with significantly high mortality. CONCLUSION: The mortality in females is significantly higher in males. The females had lower haemoglobin level and increased incidence of convulsion than their male counterparts. Shock in females has significantly higher mortality. So prompt treatment can save many lives. The increased mortality in females cannot be explained by relative inaccessibility of females to health care system as previously explained, because they have reached hospital earlier than males. Causes of shock and their association with increased mortality in females need further studies.

A case of Hymenolepis diminuta in a young male from Odisha.

Hymenolepis diminuta also known as rat tapeworm rarely causes hymenolepiasis in humans. We report a case of H. diminuta infection in an 18-year-old male farmer who presented with intermittent pruritic maculopaular rashes and dull aching left iliac fossa pain for 6 months. Patient was cured with 2 doses of praziquantel 20 mg/kg on day 0 and 7.

Onyx 18 embolisation of dural arteriovenous fistula via arterial and venous pathways: preliminary experience and evaluation of the short-term outcomes.

OBJECTIVE: This paper mainly focuses on our preliminary experience and short-term outcome evaluation of embolisation of non-cavernous dural arteriovenous fistulas (ncsDAVFs) and cavernous sinus dural arteriovenous fistulas (csDAVFs) using Onyx 18 (ev3, Plymouth, MN), and in combination with coils, via arterial and venous approaches, respectively. METHODS: Between August 2008 and March 2010, 21 DAVFs (11 ncsDAVFs and 10 csDAVFs; age range: 28-68 years; 12 females and 9 males) were undertaken. Borden classification showed Type III in 1 and Type II in 10 ncsDAVFs, and Type II in 4 and Type I in 6 csDAVFs. Onyx 18 was used in 11 ncsDAVFs (10 via single feeder and 1 via 2 feeders). Onyx 18 or in combination with coils was used in 10 csDAVFs (9 via the inferior petrosal sinus and 1 via the superior ophthalmic vein). RESULTS: Total occlusion in immediate angiography was achieved in 18 cases (85.7%; 10 ncsDAVFs and 8 csDAVFs), and near-total occlusion in 1 ncsDAVF and 2 csDAVFs. Onyx 18 was migrated into normal vasculature in two ncsDAVFs without any sequelae. One csDAVF had VI cranial nerve palsy post-operatively, which completely recovered 2 weeks post-embolisation. Follow-up angiography at 3-12 months showed complete occlusion in 20 cases (95.2%; 10 ncsDAVFs and 10 csDAVFs). One ncsDAVF (4.8%) recurred after 3 months and was successfully re-embolised. CONCLUSION: Preliminary results achieved after embolising 11 ncsDAVFs and 10 csDAVFs using Onyx 18 and in combination with coils via arterial and venous pathways, respectively, appeared to be safe, feasible and effective, as 95.2% of cases were totally occluded without any clinical sequelae.

Prevalence of methicillin-resistant Staphylococcus aureus and its antibiotic susceptibility pattern in a tertiary care hospital.

Methicillin-resistant Staphylococcus aureus (MRSA) emerged as a nosocomial pathogen in early 1960s, causing Increasing number of outbreaks in 19708, first reported in a teaching hospital in Malaysia in 1972, causing increased mortality, morbidity, and healthcare costs. Aim of this study is to screen out MRSA from various clinical samples and to see their antibiotic susceptibility pattern. From May 2008 to May 2009, 204 S aureus strains were isolated, out of which 114 (55.8%) were MRSA, and rest methicillin-sensitive Staphylococcus aureus (MSSA). Most of the MRSA strains were obtained from pus (45%) followed by urine (20.5%). Frequency of isolating MRSA were maximum in catheter tip (80%) followed by blood (66.7%) and pus (58.7%). MRSA strains were showing 100% sensitivity to vancomycin and Iinezolid, whereas 92.9% to teicoplanin. Therefore it is concluded that antibiotics other than vancomycin can be used as anti-MRSA agents after sensitivity test, as well as irrational and indiscriminate use of antibiotics can be avoided.

Management of annuloaortic ectasia in association with aortic regurgitation.

Annuloaortic ectasia associated with severe aortic regurgitation is usually managed by implanting a composite graft that comprises both valve and conduit. Synthetic valved conduits have been used by most cardiovascular surgeons, with various modifications in the technique. Yet prosthetic valves carry the risks associated with anticoagulation therapy, together with an increased risk of infective endocarditis. We report a case in which a cryopreserved aortic homograft was used for a Bentall procedure in a patient who had annuloaortic ectasia with severe aortic regurgitation. To the best of our knowledge, there are no prior published reports of homograft insertion in treatment of annuloaortic ectasia.