Clinical insights and epidemiology of central nervous system infection due to Cryptococcus neoformans/gattii species complexes: A prospective study from South India.

In the last two decades, central nervous system (CNS) cryptococcosis (CNSc) has emerged as a major opportunistic infection in the immunocompromised population of India. We have analyzed the clinical features of CNSc and epidemiology of Cryptococcus neoformans and Cryptococcus gattii. A total of 160 clinical isolates of C. neoformans/gattii recovered from CNSc patients were analyzed. The origin, clinical parameters, and imaging features of the patients were recorded, and clinical parameters were analyzed based on their human immunodeficiency virus (HIV) status and infecting species, namely, C. neoformans or C. gattii. Serotypes and mating types of the isolates were determined. Molecular typing was performed by polymerase chain reaction (PCR) fingerprinting using M13 microsatellite primer (GTG)5, and multilocus sequence typing (MLST). Majority of the patients were from Bangalore Urban, Karnataka. Among 160 cases 128 (80%) were HIV seropositive, and 32 (20%) were HIV negative. Middle-aged males (36-55 years) were highly affected. There were statistically significant differences in the clinical manifestations, imaging and CSF parameters of HIV coinfected and noninfected cases, whereas limited differences were observed in these parameters in the cases infected with C. neoformans and C. gattii. We identified 80% C. neoformans VNI, 8.75% VNII and 22.5% C. gattii (VGI), 8.75% C. tetragattii (VGIV) among clinical strains. This comprehensive study will contribute toward a better prognosis of CNS cryptococcosis patients during the hospital stay, treatment strategies for HIV coinfected and noninfected cases and will provide the molecular epidemiology of these two pathogenic fungal species in south India, which was unclear in this part of the country.

Intracranial fungal granuloma: a single-institute study of 90 cases over 18 years.

OBJECTIVE: Intracranial fungal granuloma (IFG) remains an uncommon entity. The authors report a single-institute study of 90 cases of IFG, which is the largest study until now. METHODS: In this retrospective study, all cases of IFG surgically treated in the years 2001-2018 were included. Data were obtained from the medical records and the pathology, microbiology, and radiology departments. All relevant clinical data, imaging characteristics, surgical procedure performed, perioperative findings, and follow-up data were recorded from the case files. Telephonic follow-up was also performed for a few patients to find out their current status. RESULTS: A total of 90 cases consisting of 64 males (71.1%) and 26 (28.9%) females were evaluated. The mean patient age was 40.2 years (range 1-79 years). Headache (54 patients) was the most common presenting complaint, followed by visual symptoms (35 patients), fever (21 patients), and others such as limb weakness (13 patients) or seizure (9 patients). Cranial nerve involvement was the most common sign (47 patients), followed by motor deficit (22 patients) and papilledema (7 patients). The mean duration of symptoms before presentation was 6.4 months (range 0.06-48 months). Thirty patients (33.3%) had predisposing factors like diabetes mellitus, tuberculosis, or other immunocompromised status. A pure intracranial location of the IFG was seen in 49 cases (54.4%), whereas rhinocerebral or paranasal sinus involvement was seen in 41 cases (45.6%). Open surgery, that is, craniotomy and decompression, was performed in 55 cases, endoscopic biopsy was done in 30 cases, and stereotactic biopsy was performed in 5 cases. Aspergilloma (43 patients) was the most common fungal mass, followed by zygomycosis (13 patients), chromomycosis (9 patients), cryptococcoma (7 patients), mucormycosis (5 patients), and candida infection (1 patient). In 12 cases, the exact fungal phenotype could not be identified. Follow-up was available for 69/90 patients (76.7%). The mean duration of the follow-up was 37.97 months (range 3-144 months). The mortality rate was 52.2% (36/69 patients) among the patients with available follow-up. CONCLUSIONS: A high index of suspicion for IFG should exist for patients with an immunocompromised status and diabetic patients with rhinocerebral mass lesions. Early diagnosis, aggressive surgical decompression, and a course of promptly initiated antifungal therapy are associated with a better prognosis.

Central nervous system infection due to Cryptococcus gattii sensu lato in India: Analysis of clinical features, molecular profile and antifungal susceptibility.

Cryptococcus gattii species complex has evolved as a pathogen in the last two decades causing infection among both immunocompetent and immunocompromised hosts. We aimed to analyse the clinical features of CNS infection caused by C. gattii sensu lato, molecular and antifungal susceptibility profile of this pathogen. Cases diagnosed to have CNS cryptococcosis were included in the study. Cryptococcus recovered from patient's specimen was identified by standard protocol. Species confirmation, mating type and molecular type determination were performed by PCR based methods. Antifungal susceptibility was tested in VITEK2C to amphotericin B, 5-flucytosine, fluconazole and voriconazole. Among 199 cases, 20 (10%) were due to C. gattii, comprising of 75% cryptococcal meningitis and 25% cryptococcoma cases. Young adult males were commonly affected. Headache and vomiting were prominent symptoms and 50% were immunocompromised. Among the isolates, 75%, 20% and 5% were C. tetragattii, C. gattii sensu stricto and C. bacillisporus respectively and all had mating type α. Four (20%) isolates of C. tetragattii and the only isolate of C. bacillisporus were resistant to fluconazole. The most common species isolated from south India is C. tetragattii. The study contributes to the epidemiology of C. gattii and reiterates the need for genotyping and antifungal susceptibility testing.

Significance of immune response to Mycobacterium tuberculosis culture filtrate protein antigens in cerebrospinal fluid of tuberculous meningitis patients: A search for diagnostic marker.

Mycobacterium tuberculosis (H37Ra) culture filtrate proteins (CFP) are explored as a diagnostic marker for tuberculous meningitis (TBM). Cerebrospinal fluid (CSF) samples from patients were categorized as confirmed (n = 47), suspected (n = 20), and non-TBM (n = 25) cases. Immune response by Western blot revealed TBM CSF samples are having heterogeneous response to CFP. CFP ELISA was 92% sensitive and 38.30% specific. ODs of confirmed TBM and non-TBM cases were significantly different (P < 0.0001) and also the suspected TBM and non-TBM cases (P = 0.0001). No significant difference noticed in TBM and suspected TBM (P = 0.90). Thus, CFP can be a better biomarker for the diagnosis of TBM.

Trends of CNS Cryptococcosis during Pre- and Post-HIV era: A 38 years' retrospective cohort analysis from south India.

OBJECTIVE: Central nervous system cryptococcosis (CNSc) is an AIDS defining opportunistic infection. This retrospective study aimed to analyze the changing epidemiology of CNSc cases from the period of pre- to post-emergence of HIV epidemic in south India. METHODS: Confirmed cases of CNSc from 1978 to 2015 were analyzed for demographic and clinical details with special reference to the cases diagnosed in south India during the period 1952-1977. Geographical distribution, affected age groups, clinical aspects, and comorbidities in relation to immune status were analysed RESULTS: The highest number of CNSc cases (n = 125) were recorded in 2006, with 89.6% HIV positivity. The highest HIV-positivity (93.6%) was documented in the years 2002 and 2009. CNSc cases have majorly changed after the introduction and spread of HIV in terms of predisposing factors, comorbidities, severity, affected age groups and treatment. Notably, an overall rise was observed in non-HIV associated CNSc cases from 1997 (8.1%) to 2015 (16.9%). CONCLUSION: The peak of CNSc had already reached in south India during 2005-2006. However, the number of new infections has slowly decreased in last ten years. Progressive awareness and, early diagnosis of HIV and cryptococcosis, adequate availability of HAART and potential antifungal therapy has played crucial roles in changing epidemiology of the CNSc and its associated mortality.

Cryptococcus gattii serotype-C strains isolated in Bangalore, Karnataka, India.

During a retrospective study on cryptococcosis carried out in Bangalore, Karnataka, India, four Cryptococcus gattii strains were isolated from one HIV-positive and three HIV-negative patients, two of which had unknown predisposing conditions. Serotyping and genotyping showed that the isolates were C. gattii serotype C, mating-type α and genotype VGIV. All the isolates were identical by multilocus sequence typing, but presented a low similarity compared with a set of 17 C. gattii global control strains. The comparison with a larger number of previously reported C. gattii strains, including African isolates, revealed a close relationship between Indian and African serotype-C isolates.

Advanced approach for antifungal susceptibility and characterization of resistance properties in clinical and environmental isolates of Cryptococcus species complex.

Background: Meningitis due to Cryptococcus neoformans/gattii is a fatal infection affecting immunocompromised population worldwide. Amphotericin B (AmB), fluconazole (FLC) and 5-flucytosine are the drugs of choice to treat the infection. We studied antifungal susceptibility pattern of clinical and environmental cryptococcal species using newer approach and analyze their resistant characteristics. Methods: Eighty clinical (54 C. neoformans and 26 C. gattii) and 18 environmental (14 C. neoformans and 4 C. gattii) isolates were subjected to antifungal susceptibility testing by automated (VITEK2C) method. Minimum inhibitory concentrations (MIC) were analyzed statistically. Genomic DNA of FLC resistant isolates was extracted and amplified to detect presence of CnAFR1 gene. Results: C. neoformans showed 1.85% and 21.4% AmB resistance, and 1.85% and 28.5% FLC- resistance, whereas C. gattii showed 25% and 50% FLC-resistance among clinical and environmental isolates respectively. MIC values were significantly (p < 0.05) different for the isolates from 2 sources. CnAFR1 gene sequence analysis revealed phylogenetic relationship among the resistant isolates. Conclusions: This pioneering study provides an insight into the sensitivity patterns of clinical and environmental cryptococcal isolates from south India. The recent emergence of AmB-resistance may transpire as a challenge for the clinicians. As the clinical and environmental isolates are phylogenetically evolved from CnAFR1 gene of Filobasidiella neoformans, the resistance is most probably an inherent attribute. This study emphasizes the need for speciation and antifungal susceptibility testing of cryptococcal isolates from clinical sources to institute appropriate antifungal therapy and to reduce the mortality and morbidity.

Cryptococcus gattii: A Poseur to Behold!

Cryptococcosis is an invasive systemic mycosis caused by Cryptococcus, a genus of yeast. Causative organisms for human cryptococcosis include Cryptococcus neoformans and Cryptococcus gattii. Disease due to C.neoformans is conventionally seen in patients with underlying immunosuppression, whereas C.gattii-related infection is usually seen in immunocompetent people. The fact that the infection can occur among otherwise healthy individuals underscores the importance of having a necessary understanding of the pathophysiology and clinical and radiological presentations of the disease. We report a case of disseminated pulmonary and central nervous system (CNS) cryptococcosis in an apparently immunocompetent individual with unusual radiological findings necessitating probing for alternative diagnoses. We have attempted to supplement and revise the existing data on the radiological manifestations of C.gattii.

Clinicomycological overview of brain abscess in a tertiary care center: A 38 year retrospection: Fungal brain abscess.

INTRODUCTION: Brain abscesses (BA) form approximately 8% of intracranial masses in developing and 1-2% in western countries. Fungal BA (FBA) are aggressive and represent a catastrophic manifestation compared to protozoan and bacterial BA. Diagnosis of FBA is rare and usually done postmortem. OBJECTIVES: The present retrospective study analyses the clinico-mycological aspects of FBA presented to our neurosurgical services over a period of 38 years, from January 1979 to April 2017. MATERIALS AND METHODS: Patients diagnosed as definitive cases of FBA were included in the study. Clinico- demographic and microbiological data were collected from medical records. BA pus was examined for fungal etiology using standard microbiological procedures. RESULTS: During the period of 38 years out of total 2,916 brain abscesses, 29 cases of FBA were diagnosed with an overall incidence rate of 0.99% per year. Cladophialophora bantiana (44%) was the most predominant isolate followed by Aspergillus spp and others. Male preponderance was seen with a male:female ratio of 4.8:1. There was no predilection for any age group. Headache, limb weakness and fever were the most common presentations. Amphotericin B was given in 44.8% of cases. Craniotomy with excision (48.2%) was the predominant surgical management. Outcome was fatal in 62% of the cases. CONCLUSION: Neurotropic C. bantiana is the predominant isolate causing fungal brain abscess. The incidence and trends of fungi causing brain abscess do not show significant change. Young immunocompetent outdoor working males were predominantly susceptible to fungal infection. Advance in the diagnostic modalities show promising in diagnosis of FBA. High index of suspicion with early diagnosis, prompt antifungal therapy and aggressive surgical management is required as FBA are associated with high mortality rate.

Neurobrucellosis: A neglected entity? An update from tertiary care Neurocentre of South East Asia.

Neurobrucellosis is the most serious complication of brucellosis with neither a typical clinical manifestations nor a specific cerebrospinal fluid (CSF) picture and mimics other neurological disorders leading to diagnostic dilemma. The prevalence of Neurobrucellosis ranges from 1.7 to 10% of brucellosis worldwide. This present study highlights the integrated diagnostic and clinical approaches in the diagnosis of neurobrucellosis. Cases with neurological abnormalities associated with abnormal CSF findings were included in the study. Serum and CSF samples were subjected to Rose Bengal Plate Test (RBPT), standard tube agglutination test (STAT), indirect Enzyme linked immunosorbent assay (iELISA) for IgM & IgG antibodies and polymerase chain reaction (PCR) to detect BCSP31 gene. Out of 473 cases, 278 (58.8%) were positive in serum and/or CSF by any of the methods. Out of 278, Only IgM anti-brucella antibody was positive in 105 (22.19%) cases. 122 (25.79%) cases were positive by any of the diagnostic methods in serum and not in CSF whereas 51(10.78%) cases were positive in serum and CSF and these 51 cases were considered as Neurobrucellosis among patients presenting with neurological illness. Chronic meningitis was the most common form of presentation. Multimodal differential diagnostic approaches are crucial for accurate diagnosis, effective treatment and to prevent morbidity and mortality associated with neurobrucellosis.

IgG response to Mycobacterium tuberculosis non-polar lipids and sonicated extracts among tuberculous meningitis patients.

INTRODUCTION: The diagnosis of tuberculous meningitis (TBM) is a major global health concern due to its protean nature. There is a need to identify better biomarkers for the rapid and definitive diagnosis of TBM. Lipids have been poorly explored as diagnostic markers in TBM. AIM: Non-polar lipids (NPL) and mycobacterial sonicate extract (MTSE) antigens were assessed for diagnosis of Mycobacterium tuberculosis . METHODOLOGY: A total of 110 cerebrospinal fluid samples were categorized as confirmed, suspected and non-TBM cases according to clinical presentation and laboratory investigations, which were further analysed by NPL and MTSE ELISA. RESULTS: The sensitivity and specificity of the NPL ELISA were 39.6 and 96 %, respectively, whereas the MTSE ELISA was 17 % sensitive and 92 % specific. The combination of the NPL and MTSE ELISA test was superior to these tests alone, with sensitivity and specificity of 43 and 88 %, respectively. CONCLUSION: This combination may be useful as an adjunct in the laboratory diagnosis of TBM. However, future studies in different settings among different populations, such as those with human immunodeficiency virus co-infection, are desirable to explore the full potential of biomarkers.

Case Report: Chronic Fungal Meningitis Masquerading as Tubercular Meningitis.

Phaeohyphomycosis causes a wide spectrum of systemic manifestations and can affect even the immunocompetent hosts. Involvement of the central nervous system is rare. A 48-year-old farmer presented with chronic headache, fever, and impaired vision and hearing. Serial MRIs of the brain showed enhancing exudates in the basal cisterns, and lesions in the sella and perichiasmatic and cerebellopontine angle regions along with enhancement of the cranial nerves and leptomeninges. Cerebrospinal fluid (CSF) showed lymphocytic pleocytosis with elevated protein and decreased glucose on multiple occasions. Clinical, imaging, and CSF abnormalities persisted despite treatment with antitubercular drugs and steroids for 2 years. Biopsy of the dura mater at the cervicomedullary junction revealed necrotizing granulomatous lesions, neutrophilic abscesses, and giant cells containing slender, pauci-septate, pigmented fungal hyphae. Fungal culture showed growth of Fonsecaea pedrosoi, which is classically known to cause brain abscesses. Here, we report the diagnostic odyssey in a patient with chronic meningitis from a region endemic for tuberculosis and describe the challenges in establishing the accurate diagnosis. Lack of therapeutic response to an adequate trial of empirical antitubercular therapy warrants search for alternative causes, including fungal meningitis. We highlight the uncommon manifestation of F. pedrosoi with chronic meningitis as well as the protracted clinical course despite not receiving antifungal therapy.

Case Report: Neurobrucellosis with Plastered Spinal Arachnoiditis: A Magnetic Resonance Imaging-Based Report.

Diffuse spinal arachnoiditis in neurobrucellosis is a rare manifestation. We report a boy aged 17, presenting with hearing impairment and recurrent vomiting for 18 months, weight loss for 12 months, dysphagia, dysarthria, hypophonia for 6 months, and gait unsteadiness for 5 months. He had bilateral 5th (motor) to 12th cranial nerve palsy, wasting and weakness of limbs, fasciculations, absent tendon reflexes, and positive Babinski's sign. Cerebrospinal fluid (CSF) showed raised protein and pleocytosis. Magnetic resonance imaging (MRI) showed extensive enhancing exudates in cisterns and post-contrast enhancement of bilateral 5th, 6th, 7th, and 8th nerves. Spine showed clumping with contrast enhancement of the cauda equina roots and encasement of the cord with exudates. Serum and CSF were positive for anti-Brucella antibodies. He showed significant improvement with antibiotics. At 4 months follow-up, MRI demonstrated near complete resolution of cranial and spinal arachnoiditis. It is important to recognize such rare atypical presentations of neurobrucellosis.

Intracranial Cryptococcoma-Clinicopathologic Correlation and Surgical Outcome: A Single-Institution Experience.

OBJECTIVE: To describe clinical characteristics of patients with intracranial cryptococcoma, clinicopathologic findings, and outcomes after surgery. MATERIALS AND METHODS: Clinical data were collected from hospital case records, and pathologic confirmation was done by a neuropathologist. Clinical details, imaging features, and treatment modalities were evaluated and correlated with outcomes based on regular follow-up. RESULTS: This case series included 5 cases of histology- and culture-proven intracranial cryptococcoma. Three patients (60%) were aged between the 3rd and 5th decades. Three cases were supratentorial in location, and 2 were in the posterior fossa. One patient presented with seizures. Three patients (60%) presented with fever. Three patients (60%) had papilledema, and 4 (80%) had signs of meningeal irritation. Four patients (80%) were immunocompetent. Only 1 patient was immunocompromised. All lesions were peripherally enhancing on computed tomography. Four of 5 patients underwent surgical decompression without any residue. Only 1 patient underwent stereotactic biopsy. All patients received antifungal therapy. One patient who was immunocompromised developed multiorgan failure and died after 2 months after surgery (20%). Of 5 cases, 4 (80%) had a good outcome with a mean follow-up of 5 years. CONCLUSIONS: Cryptococcus gattii is a rarer species implicated in intracranial cryptococcoma that is seen along with the more common Cryptococcus neoformans. Early diagnosis and surgical decompression followed by intravenous amphotericin B therapy for at least 6 weeks and concomitant therapy with fluconazole for prolonged periods may reduce morbidity and mortality.

Prevalence and risk factors of brucellosis among veterinary health care professionals.

The study describes prevalence, clinical symptoms and risk factors for brucellosis in personnel engaged in veterinary health care in Karnataka, India. A total of 1050 sera samples were collected from animal handlers, veterinarians, veterinary students, para-veterinarians and persons engaged in artificial insemination of animals. The sera samples were tested for brucellosis by Rose Bengal plate test (RBPT), serum agglutination test (SAT), IgG and IgM indirect ELISA and PCR. Age, sex, clinical symptoms and risk factors were recorded in structured questionnaire. Of the 1050 samples tested, 6.76, 6.38, 3.90, 2.67 and 2.0% were positive by IgG ELISA, RBPT, SAT, IgM ELISA and PCR, respectively and overall prevalence recorded was 7.04%. The prominent clinical symptoms observed were intermittent fever (71.62%) followed by joint pain and body aches. A high degree of suspicion, awareness and multimodal diagnostic approach is suggested for early diagnosis, treatment and disease follow up.

Correlation of clinical, laboratory and drug susceptibility profiles in 176 patients with culture positive TBM in a tertiary neurocare centre.

Drug resistance has increased the difficulties in control of tuberculosis infection. The present study evaluated the clinical and laboratory features among tuberculous meningitis (TBM) patients and the drug susceptibility of Mycobacterium tuberculosis (M.tb) isolated from CSF. Out of 698 CSF samples, 176 (25.21%) were M.tb culture positive. Among the clinical signs and symptoms, fever, headache and altered sensorium were found to be statistically significant (P<0.05). ELISA was a better predictor of disease and found to be statistically significant (P<0.001) in culture-proven TBM cases. Totally, 57 (32.4%) isolates were resistant to one or more drugs that include 5 (2.8%) multidrug-resistant isolates. In conclusion, the search for antibody in CSF and also CSF chloride can represent as an adjunct in the diagnosis of TBM. Screening of drug susceptibility is a very important factor and would help in better management of the disease.