Fungal Stains Research Articles

Varied presentations of complicated rhinosinusitis in COVID era: a rational approach to management

AimTo study the various presentations and manifestations of complicated rhinosinusitis in COVID era- ranging from bacterial rhino sinusitis to invasive fungal rhino sinusitis.MethodsDesign-A retrospective observational study was carried out from March 2020 to May 2021. Setting-Tertiary care hospital subjects—all COVID-positive patients who had paranasal sinus involvement. Methods-Patients were evaluated based on their symptomatology profile. Fungal stains and culture were carried out for all. They underwent Magnetic resonance Imaging and Computed Tomography scan on case-to-case basis, apart from routine nasal endoscopy. All were managed both medically and surgically depending upon their diagnosis. The natural course including outcomes, was studied, documented and analyzed.ResultsOut of 496 patients presenting with sinonasal disease, 126 were COVID-positive, 16 patients had complicated rhino sinusitis, of which 4 patients had complicated rhinosinusitis with intraorbital, intracranial or combined complications. All patients were managed successfully with combined medical and surgical approach. Twelve patients had invasive mucormycosis with overall mortality rate of 37%.ConclusionComplicated sinusitis was encountered in COVID-positive patients either when they were being actively treated for COVID-19 or as part of post-COVID sequalae. Though rhino-orbito-cerebral mucormycosis constituted the major disease burden in such patients but the possibility of bacterial rhino sinusitis with or without complications must also be kept in mind while evaluating such patients. We must remember every complicated rhinosinusitis in COVID-positive patient may not be mucor and manage appropriately.

Studded Brainstem Tuberculoma Presenting with Bilateral Inter-Nuclear Ophthalmoplegia.

Sir, A 17-year-old male with a history of low-grade fever for 2 months, headache for 15 days, and diplopia and drooping of the right upper eyelid for 10 days, was admitted to our hospital. There was no history of altered sensorium, convulsion, weakness, paresthesia, weight loss, or cough. There was no history of trauma, joint pain, rash, and high-risk behavior. His general examination was normal. Nervous system examination revealed a Glasgow Coma Scale of 15, normal visual acuity with bilateral Grade 2 papilledema, bilateral restriction of adduction movement of the eyeball with ptosis of the right eye, and dilated fixed right pupil [Figure 1a]. The rest of cranial nerve examination was unremarkable. He had bilateral upper motor neuron quadriparesis with power of 4+/5 with bilateral extensor plantars. Sensory and cerebellar functions were intact. Meningeal signs were present. Rest of the systemic examination was unremarkable.Figure 1: (a) On presentation, clinical image showing right-sided ptosis with restriction of bilateral adduction. MRI brain (i-iv) showing multiple T2 hypointense/FLAIR hyperintense signal changes in midbrain with ring enhancement in gadolinium contrast along with hydrocephalus. (b) At 1 month of follow-up, clinical image showing improvement in right-sided ptosis with reduced restriction of bilateral adduction. MRI brain (i-iv) showing reduction of midbrain lesions and resolving hydrocephalus. MRI: Magnetic resonance imaging, FLAIR: Fluid-attenuated inversion recoveryHis laboratory blood parameters showed normal hemogram with normal renal, liver, and thyroid functions with normal glucose and electrolytes. Serological examination was negative for HIV, hepatitis B and C, Leptospira, and Listeria. His cerebrospinal fluid examination revealed lymphocytic pleocytosis with elevated protein level, CBNAAT negative state, negative for Gram stain, fungal stain, and culture sensitivity. Magnetic resonance imaging (MRI) brain revealed multiple coalesced ring enhancing lesions of the midbrain with extension to upper pons with hydrocephalus [Figure 1a]. He was started on standard antitubercular treatment and corticosteroid. He showed improvement clinically within 10 days in the form of partial recovery in ptosis and adduction movements of the eyeball [Figure 1b]. Headache and fever resolved within 2 weeks. MRI brain was repeated at 1 month, which showed resolving hydrocephalus and reduction in size of the brainstem tuberculomas [Figure 1b]. The entity of the brainstem tuberculoma is a rare phenomenon in tuberculosis of central nervous system with incidence of 2.5%–8%.[1] The clinical presentation of the brainstem tuberculoma is pleomorphic. The site, size, and morphology of the lesions determine this versatility of manifestation.[2] Nuclear gaze palsy, ocular motor palsy, facial palsy, bulbar palsy, ataxia, and long tract signs are some important features of brainstem involvement.[3] A multitude of etiology can have similar presentation of brainstem encephalitis including infections, autoimmune disorders, neoplastic (gliomas), and vascular. Brainstem lesions are notorious for complications in the form of hydrocephalus, respiratory insufficiency, fatal arrhythmias, and brain herniation.[4] A thorough workup and aggressive management is prudent for correct diagnosis and better outcome. Declaration of patient consent The authors certify that they have obtained all appropriate patient consent forms. In the form, the legal guardian has given his consent for images and other clinical information to be reported in the journal. The guardian understands that names and initials will not be published and due efforts will be made to conceal identity, but anonymity cannot be guaranteed. Financial support and sponsorship Nil. Conflicts of interest There are no conflicts of interest.

Mold and Stain Resistance of Bamboo Treated with Pyraclostrobin Fungicide.

Bamboo is rich in starch and sugars and can be infected by mold and stain fungi, degrading its performance, shortening its service life, and reducing its utilization value. It is crucial to investigate how to protect bamboo against mold and stain fungi. The zone of inhibition test was used to evaluate the antifungal activity of azoxystrobin, kresoxim-methyl, pyraclostrobin and 3-iodo-2-propynyl-butylcarbamate (IPBC) against stain fungi (Botryodiplodia theobromae, Fusarium moniliforme, and Alternaria alternate) and mold fungi (Aspergillus niger, Penicillium citrinum, and Trichoderma viride) to develop new chemicals to protect bamboo against stain fungi and molds. The inhibitory activity of the composite pyraclostrobin and IPBC with different ratios was evaluated. Water-based formulations of the fungi were used to treat the bamboo, and the mold and stain resistance of the bamboo was investigated at different chemical retention rates. The results showed that the antifungal activity of pyraclostrobin was significantly higher than that of azoxystrobin and kresoxim-methyl. Different degrees of inhibitory activities against the stain and mold fungi were observed, and the inhibitory activity was higher against stain fungi than against molds. The three stain fungi were completely inhibited at a 7:3 ratio of pyraclostrobin to IPBC and 0.1% concentration. As the ratio increased, the inhibitory effect against mixed mold strains improved. The control efficacy of the pyraclostrobin formulations Str-1 and Str-2 at 0.1% concentration was 100% against Alternaria alternate and 70.8% against Fusarium moniliforme. The control efficacy of the composite formulations SI-1 and SI-2 at 0.1% concentration was 100% against all three stain fungi and greater than 91.8% against the mixed mold strains. This study provides new insights into the utilization of pyraclostrobin and its composite formulations as new bamboo antifungal agents.

Application of two fungal strains Aspergillus niger and Candida albicans in wastewater quality improvement

The nutrients Phosphorous, Nitrogen and Carbon are essential for aquatic life. However, in excess they also cause serious problems. For this reason, wastewater treatment must meet nutrient effluent limits. A wastewater sample was taken near Dhahibah village from the Erbil wastewater channel in the north of Iraq. In this experiment, pure cultures of Aspergillus niger and Candida albicans were used to treat wastewater. Samples were measured for physicochemical parameters like EC, pH, Phosphate, Nitrate, Nitrite, and BOD5 using standard methods every third day during 21 days of experiment. The results revealed that Aspergillus niger had the maximum efficiency in removing BOD5, NH4, NO3, and EC (87.27, 89.57, 83.52, and 78.49%) respectively. On the other hand, during the experimental period, Candida albicans had the maximum efficiency in decreasing PO4, and NO2, were (91.58% and 88.89%), respectively. Statistically, there were differences (P≤0.05) between the control sample and the treated wastewater sample for both fungal stains for all parameters during the experiment.

Current perspective of adrenal histoplasmosis in India: A prospective study in a tertiary care hospital, Eastern India

PurposeAdrenal histoplasmosis (AH) is an uncommon form of disseminated histoplasmosis caused by the dimorphic fungus Histoplasma capsulatum. Though, India is considered to be a non-endemic area for histoplasmosis, a high rise of AH cases is reported currently from various parts of India. Our study aimed to evaluate the current perspective of adrenal histoplasmosis in India by reviewing its clinical course, differential diagnosis, treatment, and mortality of our eleven confirmed cases of AH along with the review of authentic reported AH cases from India. Material &methodClinical materials were taken from radiologically suspected all 15 cases either with unilateral or bilateral adrenal enlargement, referred between 2018 and 2020 for microbiological investigations. Fungal stain and fungal culture along with other tests for possible differential diagnosis with AH were conducted. ResultOut of fifteen incidentaloma detected by radio-imaging, eleven cases of AH had been diagnosed in our hospital with yield of Rhodotorula spp. in one mimicking case. Nine of them were male (82%) and all were HIV nonreactive, which corroborates with the literature review. All of them had nonspecific clinical presentation of chronic abdominal pain, fever, weight loss, and anorexia. Four developed primary adrenal insufficiency, which are similar to the literature review (41%). On treatment with itraconazole and/or amphotericin B, all patients survived except one lost in follow-up. ConclusionMale preponderance and non-compromised immune status are two special characteristics of most AH though reasons are ill understood. So, mycological investigations are to be done for every such case.

Selected physical parameters of spruce chips vs. alkali consumptions in the impregnation step of kraft pulping

The uniformity of pulp and the amount of rejects are influenced by many factors, but impregnation can be the most important. The opportunities for improving impregnation of kraft white liquor start in description of the parameters affecting the process. The first parameter is the chip itself (its basic density, moisture, and pH) and the second is the process parameters set by the technological possibilities of the digester. During outside storage, the chips deteriorate via the fungal stains, brittleness, and high acidity. The moisture content also decreases. Understanding the relationships of chip parameters in the impregnation step and amount of rejects requires the assignment of effective alkali consumption. This study shows the effects of selected physical parameters, such as bulk density, moisture, and pH, of spruce chips on alkali consumption during impregnation step of a Super Batch process.

ROBOTIC-ASSISTED BRONCHOSCOPY: A POTENTIAL NEW TOOL FOR CAVITARY PULMONARY LESIONS

ROBOTIC-ASSISTED BRONCHOSCOPY: A POTENTIAL NEW TOOL FOR CAVITARY PULMONARY LESIONS

S2312 Esophageal Stricture Secondary to Mycophenolate Mofetil-Induced Injury

Introduction: Immunosuppressant medications like mycophenolate mofetil (MMF) have recently garnered more attention in the literature due to gastrointestinal toxicity. There are increasing case reports of transplant patients on MMF who develop inflammation or ulcers throughout the GI tract. It is theorized that the inflammation is due to MMF toxicity from metabolite formation. Although there has been increasing evidence of MMF induced injury to the GI tract, there are few reports of esophageal involvement, which is presented below. Case Description/Methods: This case involves a 40 year-old patient with Type I diabetes mellitus complicated by gastroparesis, renal and pancreatic transplants on immunosuppression with MMF, who developed progressively worsening dysphagia, weight loss, nausea, and vomiting. The patient initially presented to an outside gastroenterologist due to complaints of nausea and hematemesis. An EGD was completed with evidence of ulcerative esophagitis with biopsies negative for bacterial, fungal and viral stains. He was started on a proton pump inhibitor (PPI) for treatment of presumed reflux esophagitis. After no improvement with several months of therapy, he presented to our gastroenterology clinic for evaluation. Upon questioning, he specifically complained of progressive difficulty with swallowing after starting MMF. The patient underwent EGD and showed a severe distal esophageal stricture, which was not able to be traversed with the regular upper endoscope (Figure). Biopsies were taken and revealed acute erosive esophagitis. Immunohistochemical staining was negative for HSV and CMV. Given our high suspicion for MMF-induced esophageal injury, his medication was changed to liquid formulation. The patient underwent repeat serial endoscopies with dilation every 2 weeks for the next 2 months. After transitioning to liquid formulation, each subsequent endoscopy showed improvement in the ulcerated stricture. The patient was last dilated to 45 French in which he reported marked improvement of his dysphagia. Discussion: This case demonstrates a patient who developed a severe esophageal stricture from MMF induced injury with resolution of symptoms after multiple dilations and transition to a liquid formulation of MMF. Though the exact physiologic mechanisms are not well understood, gastroenterologists should be suspicious in patients on mycophenolate who present with dysphagia and stricture that is not explained by infection, malignancy, reflux, or other medications.Figure 1.: A. Initial non-traversable esophageal stricture B. Esophageal stricture status post multiple dilations C. Improving ulcerative stricture after switching to liquid formulation of MMF and multiple dilations.

S3034 Histoplasmosis Infection Presented as Cholestatic Liver Disease

Introduction: Histoplasmosis is the most common mycosis in the United States with peak incidence in the Midwest region. In ∼21% of patients with systemic histoplasmosis hyperbilirubinemia can be detected. However, cholestatic presentation of histoplasmosis infection is very rare. Here we present a case of immunocompromised patient with predominant cholestatic liver disease caused by Histoplasma Capsulatum. Case Description/Methods: This is the case of 56- years- old female who presented with– sided abdominal pain, shortness of breath, chills, hot flushes, nausea, vomiting and diarrhea. She is suffering from rheumatoid arthritis. Her medications are adalimumab, celecoxib, hydroxychloroquine. She denied alcohol intake. Her initial workup including liver function tests (LFTs) was positive for a total bilirubin 3.5 mg/dl, conjugated bilirubin 2.7 mg/dl, alkaline phosphatase – 547 U/L, AST – 152 U/L, ALT-142 U/L, and platelet count – 55 k/ul. During hospitalization her abdominal pain was getting progressively worse, and her LFTs continued to trend up. Eventually liver biopsy was performed. The results showed multiple granulomas with central necrosis, and associated portal and lobular inflammation. The fungal stain highlighted yeasts suggestive of histoplasma. Her urinary histoplasma Ag Quantitative EIA eventually came back positive -8.87 ng/ml. Viewing the normal chest x-ray and ongoing hypoxia, chest CT scan with IV contrast was performed showing 1.1 cm lung nodule in the left lower lobe. A subsequent bronchoscopy revealed budding yeasts. Patient was started on liposomal amphotericin B, and her LFTs trended down. The patient was treated with amphotericin B for 21days subsequently changed to twice-daily oral Itraconazole 200mg. At the outpatient follow up visit, patient admitted a complete resolution of her abdominal pain and improvement of her breathing status. Discussion: Liver histoplasmosis is a rare cause for cholestatic liver disease. Over the past two decades, a limited number of cases with similar clinical scenario were described. Despite of the rarity of this condition, hepatic histoplasmosis should be considered in a selected population of immunocompromised patients living in an endemic area (ex: Ohio). Inclusion of urine antigen testing for histoplasma in early workup can potentially prevent diagnostic delay.

S2014 Disseminated Histoplasmosis Presenting as Right Colon Lesions in HIV-Negative Patients: Report of Two Cases

Introduction: Histoplasma capsulatum is the most common endemic mycosis in the gastrointestinal tract of immunocompromised hosts. However, it can also present in immunocompetent patients. Terminal ileum and cecum are most commonly affected, recognized on endoscopic examination as inflammation, ulcerated lesions, or polypoid masses. In most patients, gastrointestinal manifestation takes a subclinical disease course. We present 2 cases of immunocompetent patients with colon lesions attributed to disseminated histoplasmosis. Case Description/Methods: First case is a 62-year-old African American woman admitted to the hospital due to abdominal pain, weight loss and poor oral intake. She was found to be anemic. CT revealed a large retroperitoneal mass with central necrosis suspicious for malignancy. Colonoscopy revealed a cecal ulcer (Figure A) with pathology reporting mucosal ulceration associated with granulomatous inflammation. GMS and PAS fungal stain showed small ovoid yeast forms, morphologically compatible with histoplasmosis (Figure B).Biopsy of the retroperitoneal mass was also positive for histoplasma. Second case is a 54-year-old Hispanic man admitted to the hospital with fluid overload secondary to acute renal failure attributed to post-infectious glomerulonephritis. He was also found to have anemia. Colonoscopy showed a 6 mm polyp in the ascending colon removed by cold snare technique (Figure C). Pathology reported polyp fragment with macrophages and numerous 2-4 micron non-budding yeast forms best seen on GMS stain, most consistent with histoplasmosis. CT scan showed prominent retroperitoneal lymphadenopathy. Both our patients were started in itraconazole and are being closely followed by the ID service. Discussion: Disseminated histoplasmosis is usually seen in immunocompromised patients such as those with HIV and medication-induced immunosuppression. GI involvement and symptoms can be diverse. Our first patient presented with a large retroperitoneal mass and cecal ulcer causing her abdominal discomfort. Second patient was much more subtle as histoplasmosis presented as a polypoid lesion. Both patients presented with significant anemia, which prompted endoscopic evaluation. Although the diagnosis of histoplasmosis in immunocompetent patients usually requires a high index of suspicion, gastroenterologists should be familiar with its variable presentations in the GI tract.Figure 1.: a) Histoplasma presenting as cecal ulcer. b) GMS stain showing Histoplasma. c) Polypoid lesion positive for Histoplasma.

P416 Efficient and accurate diagnosis of otomycosis using an ensemble deep learning model

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PMObjectivesOtomycosis accounts for ˃15% of the cases with external otitis worldwide. And otomycosis is more frequently observed in humid regions and people enjoying the culture of ear cleaning in China. Aspergillus and Candida are the major pathogens that could cause long-term infection. Early endoscopic and microbiological examinations are important for appropriate medical treatment to otomycosis. However, accurate diagnosis always needs experts such as otologist and microbiologist. Deep learning model is a novel efficient method to provide quick diagnosis which is an automatically diagnostic program using a large database of images acquired in the cliniC. This paper puts forward a mechanic learning model to address the diagnosis of otomycosis caused by Aspergillus and Candida accurately and quickly.MethodsWe proposed a computer-aided decision system that is based on a deep learning model consisting of two subsystems, a java-based web application, and picture classification. The web application subsystem mainly provides a user-friendly page for collecting consulted pictures as well as displaying the calculation results. The picture classification subsystem mainly uses trained neural network models for end-to-end data inference. The end user only needs to upload a few pictures of the ear endoscope, and the system will return the classification results to the user in the form of category probability value.In order to accurately diagnose otomycosis, we generally kept endoscopic images and took the secretion for fungal culture for further identification. Positive fluorescence fungal staining, culture, and further DNA sequencing were taken to confirm the pathogens, Aspergillus or Candida sp. In addition, impacted cerumen, external otitis, and normal external auditory canal endoscopic images are retained for reference. We merged these four types of images into an endoscopic images gallery.ResultsIn order to achieve better accuracy and generalization ability after model training, we selected 2750 samples from nearly 4000 ear endoscopic images as training samples and 454 as validation samples. On the selection of deep neural network models, we tested the resnet, senet, and efficientnet neural network models with different numbers of layers. Considering the accuracy and operation speed, we finally chose the efficientnet-b6 model and output the probability values of the four categories of otomycosis, external otitis, impacted cerumen, and normal cases. After multiple iterative sample training, the overall validation sample accuracy reached 94.71%, and the average cross-validation accuracy of the 4 classifications reached 94.3%.ConclusionThe results suggest that the system can be used as a reference for general practitioners to make better decisions in the diagnosis of otomycosis.

P491 Scedosporium Apiospermum brain abscess in an immunocompetent host: Rare case from Southern India

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PMObjectives/Introduction Scedosporium apiospermum is a filamentous fungus that causes a broad spectrum of diseases in an immunocompromised host involving the lungs, skin, bones, eyes, joints, and the central nervous system. It is a rare cause of fungal brain abscess, more so in an immunocompetent individual. Here, we report a case of brain abscess in an immunocompetent host caused by S. apiospermum.MethodsA 78-year-old retired railway officer from Chennai, presented to our hospital on May 4, 2022, with a 3-month history of weakness, gait instability followed by difficulty in walking, and left hemiparesis. In all, 20 days before the presentation, he had an episode of generalized tonic-clonic seizure with worsening of his neurological state resulting in bed-bound status. His medical history included Pemphigus vulgaris for which he was on topical treatment, well-controlled diabetes mellitus, hypertension, and coronary artery disease.He underwent an MRI brain which revealed a T2 heterointense intra-axial right parietal lesion with significant perilesional edema. A provisional diagnosis of a cerebral abscess and malignancy was entertained. He was subjected to craniotomy and the surgical findings were consistent with a brain abscess and the pus was evacuated and sent for microbiological analysis. The pus fungal stain was consistent with septate hyphae and the cultures grew S. apiospermum. The histopathological findings were also consistent with a brain abscess caused by septate fungi (Aspergillus like fungi).Results/TreatmentHe was started on Liposomal Amphotericin B 5 mg/kg dose IV OD and injectable voriconazole. There was an initial clinical improvement with respect to sensorium and neurological status. He subsequently developed bradycardia with worsening sensorium to which he finally succumbed.Conclusion Scedosporium apiospermum is an asexual form of Pseudoallescheria boydii, a fungus found in soil, contaminated water, and sewage. It is a rare cause of brain abscess in immunocompromised individuals. Near drowning or trauma may be the causative factors for immunocompetent individuals. Our patient was a well-controlled diabetic host with no apparent immunosuppression. Scedosporium apiospermum is diagnosed on the basis of culture and microbiological examination. Due to the similarities in the clinical and histopathological presentation of Scedosporium with other similar fungi, culture becomes the gold standard tool for diagnosis.Treatment includes surgical drainage of the abscess along with intravenous voriconazole for at least 8-12 weeks. The prognosis depends upon the immune status, surgical intervention, and medical antifungal therapy.

P16.27: A Case of Cytomegalovirus (CMV) Esophagitis Without CMV Viremia After SARS-CoV-2 Infection in a Kidney Transplant Recipient While on CMV Prophylaxis

Introduction: CMV prophylaxis significantly reduces CMV infection after solid organ transplants, but patient may still present with CMV primary disease or reactivation during the first six months post-transplant. Although high CMV viral load is usually seen in tissue invasive disease, blood CMV PCR my not be able to diagnose gastrointestinal CMV disease as sensitivity of blood CMV PCR is lower in seropositive patients. CMV and SARS-CoV-2 co-infection has been reported particularly in elderly and immunocompromised patients. We report a unique case of CMV esophagitis without viremia diagnosed after recent SARS-CoV-2 infection in a kidney transplant recipient. Case: Twenty-eight year old Caucasian female with end stage kidney disease due to unknown etiology received negative crossmatch living related renal transplant from her sister. She received Basiliximab for induction and Tacrolimus and mycophenolic acid as maintenance immunosuppression. Patient had intermediate risk for CMV (Donor CMV IgG negative and recipient IgG positive) and was on Valganciclovir for prophylaxis as per the protocol. She had a stable allograft function with serum creatinine around 1-1.2 mg/dl. She developed SARS-CoV-2 infection with mild respiratory symptoms after 2 months of kidney transplant. Mycophenolic acid was held very briefly, and Prednisone 10 mg daily was added to the regimen. She reported chest pain, nausea and vomiting after resuming mycophenolic acid. Blood CMV PCR was undetectable and other laboratory studies were significant for anemia of 9.3 g/dl (baseline 10 g/dl), with stable renal and hepatic functions, without leucopenia or thrombocytopenia. Upper Gastrointestinal endoscopy revealed a large 4-5 cm circumferential ulceration in the mid esophagus with biopsy features suggesting viral cytopathic effect and rare CMV infected cells. Stomach and duodenum were normal. HSV and fungal stains were negative. Patient was started on oral valganciclovir induction dose and showed symptomatic recovery in a few weeks. Conclusion: High index of suspicion is necessary to diagnose CMV co-infection and/or reactivation in the settings of SARS-CoV-2 infection and the tissue biopsy is the key for diagnosis of the tissue invasive gastrointestinal disease.

Antimicrobial activities of polyethylene glycol and citric acid coated graphene oxide-NPs synthesized via Hummer’s method

Graphene oxide (GO)-NPs possess excellent physicochemical and biological properties and could have prominent antimicrobial activities. We used solution-based Hummer’s method to synthesize pure, CA coated and PEG coated GO-NPs and the XRD revealed hexagonal lattice structure and the crystallite size of pure and coated GO-NPs calculated in the range of 7–16 nm. In addition, a spongy like surface in pure GO-NPs and randomly crumpled like surface by using the CA and PEG coating agents was also identified and SEM images irregular, non-uniform and rod shape morphology of the graphite powder. The GO in the UV region and the band gap decreased from 2.30 to 2.03 eV while different modes (C C and C O) on the surface of pure and coated GO-NPs were evident as indicated by Raman spectra. Finally, in-vitro antimicrobial activity was assessed against two-gram negative bacterial strains viz. A. hydrophila and E . coli and one fungal stain F. avenaceum at 10 and 20 mg/mL concentrations. Our results indicated enhancement in the inhibition zones of pure GO-NPs from 8 to 17 mm, PEG coated GO-NPs from 8 to 20 mm and CA coated GO-NPs from 8 to 17 mm respectively. Overall the PEG coated GO-NPs had prominent antimicrobial activities and we recommend application of polymer coated GO-NPs for wound healing process.

Pan-cancer analyses reveal cancer-type-specific fungal ecologies and bacteriome interactions

SummaryCancer-microbe associations have been explored for centuries, but cancer-associated fungi have rarely been examined. Here, we comprehensively characterize the cancer mycobiome within 17,401 patient tissue, blood, and plasma samples across 35 cancer types in four independent cohorts. We report fungal DNA and cells at low abundances across many major human cancers, with differences in community compositions that differ among cancer types, even when accounting for technical background. Fungal histological staining of tissue microarrays supported intratumoral presence and frequent spatial association with cancer cells and macrophages. Comparing intratumoral fungal communities with matched bacteriomes and immunomes revealed co-occurring bi-domain ecologies, often with permissive, rather than competitive, microenvironments and distinct immune responses. Clinically focused assessments suggested prognostic and diagnostic capacities of the tissue and plasma mycobiomes, even in stage I cancers, and synergistic predictive performance with bacteriomes.

Rapid Detection of Candida tropicalis in Clinical Samples From Different Sources Using RPA-LFS.

Candida tropicalis is one of the few Candida species besides Candida albicans that is able to produce true hyphae. At present, the commonly used clinical methods for the identification of this organism are traditional fungal culture, CTB staining, and color development. Polymerase chain reaction (PCR) and real-time quantitative PCR (qPCR) are also used to identify this fungus. Since the course of C. tropicalis infection progresses rapidly, there is an urgent need for rapid, sensitive, real-time field assays to meet the needs of clinical diagnosis. Recombinase polymerase amplification (RPA) combined with lateral flow strip (LFS) can rapidly amplify and visualize target genes within 20 min, and by pre-processing samples from different sources, the entire process can be controlled within 30 min. In this study, RPA-LFS was used to amplify the internal transcribed spacer-2 (ITS2) gene of C. tropicalis, and primer-probe design was optimized by introducing base mismatches to obtain a specific and sensitive primer-probe combination for clinical sample detection. LFS assay for 37 common clinical pathogens was performed, sensitivity and specificity of the detection system was determined, reaction temperature and time were optimized, and 191 actual clinical samples collected from different sources were tested to evaluate the detection performance of the established RPA-LFS system to provide a reliable molecular diagnostic method for the detection of C. tropicalis, the results show that the RPA-LFS system can specifically detect C. tropicalis without cross-reacting with other fungi or bacterial, with a sensitivity of 9.94 CFU/µL, without interference from genomic DNA of other species, at an optimal reaction temperature of 39°C, and the whole reaction process can be controlled within 20 min, and to meet the clinical need for rapid, sensitive, real-time, and portable field testing.

CLINICO-MICROBIOLOGICAL STUDY OF PATIENTS WITH ACUTE EXACERBATION OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE

Background: Chronic obstructive pulmonary disease is the third leading cause of death worldwide. COPD patients are frequently colonised with potential respiratory pathogens. Use of empirical antibiotics without proper investigations in acute exacerbation leads to emerging resistant strains. Aim: This study is aimed to record the clinical profile of patients with acute exacerbation of COPD with special reference to microbiological profile so that early adequate treatment can be given. Material and Methods: This observational cross sectional study was conducted on 104 patients with acute exacerbation of COPD admitted in Medicine department of Jorhat Medical college and hospital from 1st July 2020 to 30th June 2021.After written informed consent detailed history,spirometry, sputum culture and sensitivity with AFB, Gram and fungal staining, chest X- ray, blood parameters and ECG were done. Result: COPD is common above 40 years of age with male predominance (63.46%).36.54% were active smoker,25% were ex smoker and 38.46% were non smoker. Positive bacteriological culture was obtained in 32.69% of cases, commonest organism being Klebsiella pneumoniae (24%)followed by Pseudomonas aeruginosa (1.92%), Escherichia coli.(0.96%)and Citrobacter freundii(0.96%).32% of Klebsiella pneumoniae,40% of Klebsiella oxytoca and 100% of Citrobacter were found to be multidrug resistant strain. Conclusion : Gram negative bacteria were more frequently isolated in our study. Early antimicrobial treatment reduces the increasing burden of COPD.

Histomorphological features of mucormycosis with rise and fall of COVID-19 pandemic

IntroductionDue to Corona Virus disease -19, India saw a surge of mucormycosis cases, associated with high death rate. India, during the month of May to July 2021 saw a surge of mucormycosis from all states, with close to 50,000 cases just in a span of 3 months. ObjectiveTo examine the histopathological appearances of rhino-orbital/rhino-maxillary/sino-nasal mucormycosis in the backdrop of the ongoing COVID 19 pandemic. Material and methodsThe study involved analysis of 60 biopsy samples of suspected rhino-maxillary /rhino-orbital mucormycosis received from post-COVID-19 patients. A preliminary review of the slides showing hyphal forms of fungal organisms with un-doubtful tissue / mucosal invasion was included. All samples were examined under Hematoxylin and Eosin stains along with special fungal stains. Data thus obtained were analyzed statistically. Special stains for fungus namely Periodic Acidic Schiff (PAS) and Gomori Methenamine silver (GMS) were utilized to confirm and/or to differentiate the fungal organisms and to highlight the cell wall of the fungus. ResultsThe mean age of the patients with mucormycosis was 51.68 years and 72 (83.33%) of them were males. Acute type of inflammation was noted in 44 (73.33%), granulomatous inflammation in 14 (23.33%) of cases. Bony invasion and perineural invasion was observed in 5 (8.33%) and 55 (91.67%) cases, respectively. The dominant fungus were mucorales in 58 (96.67%), aspergillous, along with mucorales in 12 (20%) and combination of mucorales and candida identified in 8 (13.33%) cases. ConclusionBesides all the histological appearance of angioinvasion, bone, and soft tissue invasion, a notable aspect was the shift in inflammatory pattern, which was more granulomatous in nature, with a decrease in fungal load correlating with the drop of COVID second wave. This proves that as immunity develops, the host's response to secondary opportunistic infections changes.

Pewarnaan Alternatif dengan Menggunakan Filtrat Kulit Kenari pada Uji Mikroskopik Jamur Candida Albicans dan Aspergillus Niger

Abstract. Walnut is a native plant of Indonesia that grows a lot in eastern Indonesia, such as North Sulawesi, Maluku and the island of Seram. Walnut shells contain phenolic compounds which are natural antioxidants. Phenolic compounds are used as preservatives because phenol can scavenge free radicals. The color contained by walnut shells may be an alternative to fungal microscopic test staining. This study aims to see the fungus on alternative coloring of walnut shells by using a microscopic test. The method used in this research is descriptive with an experimental approach. The results of the study identified as many as 3 samples (100%) Candida albicans and 3 samples (100%) Aspergillus niger in the filtrate by heating treatment, as well as 3 samples (100%) Candida albicans and 3 samples (100%) Aspergillus niger in the filtrate with treatment without heating. Microscopic observations of Candida albicans and Aspergillus niger fungi, appeared more clearly and stained with the use of alternative coloring filtrate of walnut shell filtrate with the heating process. Conclusion : Alternative staining with walnut shell filtrate can be used to color Candida albicans and Aspergillus niger fungi. The filtrate by heating is more effective in the microscopic examination of fungi. Keywords: walnut shell, alternative coloring, candida albicans, aspergillus niger

Comparison of Fungal Fluorescent Staining and ITS rDNA PCR-based Sequencing with Conventional Methods for Diagnosis of Onychomycosis

Accurate identification of fungal causes for onychomycosis is essential for proper treatment. Presently available laboratory methods show unreliable sensitivity; so there is a requirement for innovative detection techniques. The aim for this work was to assess the efficiencies of fluorescent staining and internal transcribed spacer (ITS) ribosomal DNA (rDNA) polymerase chain reaction (PCR)-based sequencing in comparison to conventional techniques for diagnosis of onychomycosis. Nail specimens obtained from 100 patients with clinically- diagnosed onychomycosis were analyzed. Nail scrapings or clippings were subjected to direct microscopic examination by KOH mount, culture by using Sabouraud’s dextrose agar and histopathological examination with periodic-acid Schiff (PAS). Collected specimens were subsequently examined by fluorescent staining and PCR-based sequencing (30 specimens only) to compare the feasibility, sensitivity and diagnostic accuracy for these two methods. The most frequently isolated fungi were yeasts (39/76: 51.3%), dermatophytes (24/76; 31.6%) and non-dermatophyte molds (NDMs) (13/76; 17.1%). Mixed mycotic infections were recovered from 6% of the collected nail specimens. The positive detection rates were significantly different between KOH examinations (52%), nail plate histology (55%), fungal culture (70%) and fluorescent staining (80%). Considering fungal culture as the gold standard, the most sensitive technique was PCR (100%) followed by fluorescent staining (89%), PAS staining (69%) while the least sensitive technique was KOH mount (53%). Fluorescence staining can be used as a rapid and high-yield technique for identification of fungi in the specimens. PCR-based sequencing was highly sensitive and faster compared to culture. Whenever possible, it enables species identification with higher adequacy.