Aspergillus Hyphae Research Articles

Characterisation of the spatio-temporal localisation of a pan-Mucorales specific antigen during germination and immunohistochemistry.

Mucormycosis is an aggressive, invasive fungal infection caused by moulds in the order Mucorales. Early diagnosis is key to improving patient prognosis, yet relies on insensitive culture or non-specific histopathology. A pan-Mucorales specific monoclonal antibody (mAb), TG11, was recently developed. Here, we investigate the spatio-temporal localisation of the antigen and specificity of the mAb for immunohistochemistry. We use immunofluorescence (IF) microscopy to assess antigen localisation in eleven Mucorales species of clinical importance and live imaging of Rhizopus arrhizus germination. Immunogold transmission electron microscopy (immunoTEM) reveals the sub-cellular location of mAb TG11 binding. Finally, we perform immunohistochemistry of R. arrhizus in an ex vivo murine lung infection model alongside lung infection by Aspergillus fumigatus. IF revealed TG11 antigen production at the emerging hyphal tip and along the length of growing hyphae in all Mucorales except Sakasenea. Timelapse imaging revealed early antigen exposure during spore germination and along the growing hypha. ImmunoTEM confirmed mAb TG11 binding to the hyphal cell wall only. The TG11 mAb specifically stained Mucorales but not Aspergillus hyphae in infected murine lung tissue. TG11 detects early hyphal growth and has valuable potential for diagnosing mucormycosis by enhancing discriminatory detection of Mucorales in tissue.

BTK drives neutrophil activation for sterilizing antifungal immunity.

We describe a previously-unappreciated role for Bruton's tyrosine kinase (BTK) in fungal immune surveillance against aspergillosis, an unforeseen complication of BTK inhibitors (BTKi) used for treating B-cell lymphoid malignancies. We studied BTK-dependent fungal responses in neutrophils from diverse populations, including healthy donors, BTKi-treated patients, and X-linked agammaglobulinemia patients. Upon fungal exposure, BTK was activated in human neutrophils in a TLR2-, Dectin-1-, and FcγR-dependent manner, triggering the oxidative burst. BTK inhibition selectively impeded neutrophil-mediated damage to Aspergillus hyphae, primary granule release, and the fungus-induced oxidative burst by abrogating NADPH oxidase subunit p40phox and GTPase RAC2 activation. Moreover, neutrophil-specific Btk deletion in mice enhanced aspergillosis susceptibility by impairing neutrophil function, not recruitment or lifespan. Conversely, GM-CSF partially mitigated these deficits by enhancing p47phox activation. Our findings underline the crucial role of BTK signaling in neutrophils for antifungal immunity and provide a rationale for GM-CSF use to offset these deficits in susceptible patients.

A rare case of meningoencephalitis due to <i>Aspergillus fumigatus</i>

Cerebral Aspergillosis (CA) is an opportunistic fungal infection that usually affects immunocompromised hosts. A man was admitted for loss of consciousness and aphasia. Computed Tomography (CT) scan showed a left cerebral frontal lesion. Nasal swab showed hyphae of Aspergillus spp.; isavuconazole was started. Lumbar Puncture (LP) tested positive for galactomannan (index 1.9), and Aspergillus fumigatus grew on the Cerebrospinal Fluid (CSF) culture. Unfortunately, a new episode of fever and a general deterioration then ensued up to the death. In immunocompetent patients as well, chronic fungal rhinosinusitis deserves attention due to its potential for rapid evolution to Central Nervous System (CNS) involvement.

Determination of aspergillosis of the female genital organs in women with immunodeficiency

Background. Morphological diagnosis of aspergillosis sometimes poses great difficulties due to the fact that molds of the genus Aspergillus are very sensitive to the use of antimicrobial therapy and change under its influence. In this regard, we provide information about aspergillosis of the female genital organs, discovered accidentally during an autopsy. The purpose of the study is to conduct a complex pathomorphological analysis of biopsy, operative and sectional gynecological material of women with secondary immunodeficiency conditions, for early diagnosis of aspergillosis (which is the key to a favorable outcome of the disease), differential diagnosis and detection of invasive forms of aspergillosis. Materials & Methods. We analyzed the autopsies of 4,796 women who died in Odesa region from 2009 to 2022. Autopsies revealed 7 cases of aspergillosis with damage to the female reproductive system. All of them were found in women who had confirmed HIV infection. In 5 cases, women additionally suffered from chronic alcoholism. Results. Based on the results of these cases, it can be stated that the detection of characteristic mycelium in the preparations is a reliable sign of aspergillosis, but their absence does not deny this disease. Aspergillus hyphae change as the process develops: they can be weakly stained, and with dystrophic phenomena and the presence of necrotic fragments of mycelium, it can be almost colorless when using hematoxylin and eosin. While necrosis, tissue detritus is observed in the center of the granuloma, fibrous structures, fungal hyphae are formed perifocally. To confirm the diagnosis, it is important to detect granulomatous productive inflammation, conidial heads, inflammatory infiltration, microabscesses, foci of calcifications. However, against the background of immunodeficiency, it is not always possible to see the "classic morphological picture" of aspergillosis. In addition, detection of characteristic mycelium is possible only in a proportion of patients. Conclusions. When examining the autopsy material of deceased women with immunodeficiency states, attention should be paid to the specific etiology of inflammatory processes of the reproductive system and the nature of the inflammatory process, the presence of Aspergillus hyphae and mycelia. The presence of morphological changes characteristic of aspergillosis allows to conduct a diagnostic search in the right direction. Detection of an invasive form of aspergillosis may indicate the presence of an immunodeficient state. Keywords: aspergillosis, female reproductive system, immunodeficiency state, HIV infection, uterus, mammary gland, drug addiction.

Secondary cutaneous aspergillosis in a child with Behçet’s disease: a case-based update

BackgroundInvasive aspergillosis (IA) is one of the rarest opportunistic fungal infections and has increased in frequency worldwide in recent years. It is a life-threatening infection associated with high mortality rates. Invasive pulmonary aspergillosis (IPA) is the most severe form of the disease. Extrapulmonary forms can develop as a primary infection or occur as part of a disseminated infection from the lung in severely immunocompromised patients. The major limitation in the management of these infections is the challenge of early diagnosis.Case presentationHere we report a case of secondary cutaneous aspergillosis that developed from extensive pulmonary aspergillosis in a 3-year-old female who underwent immunosuppressive therapy for a diagnosed Behçet disease (BD). Aspergillus hyphae were identified on skin biopsies. Cultures grew Aspergillus fumigatus. The diagnosis of cutaneous aspergillosis enabled us to diagnose IPA, although there was no mycopathological proof of lung infection. The patient was successfully treated with voriconazole (8 mg/kg/day) and surgical debridement of the skin lesion.ConclusionsAlthough cutaneous involvement in aspergillosis is extremely uncommon, it may be the presenting feature in some cases, allowing for an accurate and timely diagnosis of deeply infected sites. Accordingly, when evaluating skin lesions in immunocompromised individuals, especially debilitated children with underlying diseases requiring long-term immunosuppressive agents, cutaneous aspergillosis should be vigilantly considered.

Innate and Adaptive Immune Responses Induced by Aspergillus fumigatus Conidia and Hyphae.

Previous research indicated that hyphae of Aspergillus fumigatus (A. fumigatus) rather than conidia could successfully build a pulmonary aspergillosis model in immunocompetent mice. In this study, we compared the immune responses induced by hyphae and conidia to explore the possible mechanism of this striking phenomenon. Herein, a novel method was designed and adopted to quantify hyphal fragments. Murine macrophages RAW264.7 and human peripheral blood mononuclear cells were stimulated by A. fumigatus hyphae and conidia in vitro, respectively, and then immunological reactions were measured. Male C57BL/6 mice were challenged with conidia and hyphae through intratracheal inoculation. Dynamic conditions of mice were recorded, and RNA-seq measured corresponding immune responses. The results of the study confirmed that hyphae could induce more intensive inflammation than conidia in vitro and in vivo. However, macrophages revealed a higher production of ROS and M1 polarisation in response to conidia stimuli. Additionally, conidia could promote Th1 cell differentiation, while hyphae could increase the CD4/CD8 ratio. RNA-seq validated the fact that those multiple immunologically relevant pathways were more strongly activated by hyphae than conidia, which also promoted Th2 cell differentiation and suppressed Th1 signalling. Both hyphae and conidia could activate Th17 signalling. In general, conidia and hyphae induced distinctly different host immune responses, and the immune responses induced by conidia played a better protective effect. Therefore, the unique function of hyphae in the spread and infection of Aspergillus should be emphasised, and more research is required to clarify the underlying mechanisms for better understanding and management of aspergillosis.

A Novel Monoclonal Antibody 1D2 That Broadly Inhibits Clinically Important Aspergillus Species.

Aspergillus fumigatus is a ubiquitous airborne fungus, is the predominant cause (>90%) of invasive aspergillosis (IA) in immunosuppressed patients and has a high mortality. New approaches to prevention and treatment are needed because of the poor efficacy, toxicity and side effects of the current anti-Aspergillus drugs on patients. Thus, we aim to explore a new avenue to combat Aspergillus infection by using a novel monoclonal antibody (mAb) 1D2 against a glycoprotein on the cell wall of Aspergillus. The ability of this mAb to inhibit attachment, germination, and growth of Aspergillus conidia and hyphae in vitro were examined. A dose-dependent growth inhibition of Aspergillus conidia in the presence of mAb 1D2 was found. The mAb 1D2 inhibited attachment of Aspergillus conidia to an untreated slide surface and fibronectin-treated surface compared to an unrelated mAb 6B10. When conidia were exposed to 1D2 concomitantly with inoculation into culture media, the mAb prevented the swelling and germination of conidia. This inhibitory ability of 1D2 was less apparent if it was added two hours after inoculation. Damage to hyphae was also observed when 1D2 was added to Aspergillus hyphae that had been incubated in media overnight. These in vitro results indicate that mAb 1D2 broadly inhibits clinically important Aspergillus species and has a promising therapeutic effect both as prophylaxis to inhibit an Aspergillus infection as well as a treatment.

Subcutaneous canine fossa aspergilloma to intervene or not - case report

<p>The term “Aspergillosis” refers to an illness due to allergy, airway or lung invasion, cutaneous infection, or extrapulmonary dissemination caused by species of <em>Aspergillus</em>. A 51-year-old male with a history of swelling in the right maxillary region is presented. The diagnosis was subcutaneous aspergillosis in an immunocompromised patient, which was successfully treated with voriconazole and surgical excision of the mass. Possible clinical manifestations, diagnostic imaging techniques and treatment used are highlighted upon. Patient was taken up for premaxillary mass excision by sublabial approach. Sublabial incision was given in the upper gingivo-buccal sulcus from right canine upto the first molar, the canine fossa and a subcutaneous mass was dissected from anterior wall of maxilla Histopathological report was consistent with fungal granuloma with predominant hyphae of aspergillus. Patient was prescribed voriconazole 200 mg twice a day with weekly monitoring of liver functions.</p>

Lung epithelial and myeloid innate immunity in influenza-associated or COVID-19-associated pulmonary aspergillosis: an observational study

BackgroundInfluenza-associated pulmonary aspergillosis (IAPA) and COVID-19-associated pulmonary aspergillosis (CAPA) affect about 15% of critically ill patients with influenza or COVID-19, respectively. These viral–fungal coinfections are difficult to diagnose and are associated with increased mortality, but data on their pathophysiology are scarce. We aimed to explore the role of lung epithelial and myeloid innate immunity in patients with IAPA or CAPA.MethodsIn this observational study, we retrospectively recruited patients who had been admitted to the intensive care unit (ICU) of University Hospitals Leuven, Belgium, requiring non-invasive or invasive ventilation because of severe influenza or COVID-19, with or without aspergillosis, between Jan 1, 2011, and March 31, 2021, whose bronchoalveolar lavage samples were available at the hospital biobank. Additionally, biobanked in vivo tracheobronchial biopsy samples from patients with IAPA or CAPA and invasive Aspergillus tracheobronchitis admitted to ICUs requiring invasive ventilation between the same dates were collected from University Hospitals Leuven, Hospital Network Antwerp (Belgium), and Amiens-Picardie University Hospital (France). We did nCounter gene expression analysis of 755 genes linked to myeloid innate immunity and protein analysis of 47 cytokines, chemokines, and growth factors on the bronchoalveolar lavage samples. Gene expression data were used to infer cell fractions by use of CIBERSORTx, to perform hypergeometric enrichment pathway analysis and gene set enrichment analysis, and to calculate pathway module scores for the IL-1β, TNF-α, type I IFN, and type II IFN (IFNγ) pathways. We did RNAScope targeting influenza virus or SARS-CoV-2 RNA and GeoMx spatial transcriptomics on the tracheobronchial biopsy samples.FindingsBiobanked bronchoalveolar lavage samples were retrieved from 166 eligible patients, of whom 40 had IAPA, 52 had influenza without aspergillosis, 33 had CAPA, and 41 had COVID-19 without aspergillosis. We did nCounter gene expression analysis on bronchoalveolar lavage samples from 134 patients, protein analysis on samples from 162 patients, and both types of analysis on samples from 130 patients. We performed RNAScope and spatial transcriptomics on the tracheobronchial biopsy samples from two patients with IAPA plus invasive Aspergillus tracheobronchitis and two patients with CAPA plus invasive Aspergillus tracheobronchitis. We observed a downregulation of genes associated with antifungal effector functions in patients with IAPA and, to a lesser extent, in patients with CAPA. We found a downregulated expression of several genes encoding proteins with functions in the opsonisation, recognition, and killing of conidia in patients with IAPA versus influenza only and in patients with CAPA versus COVID-19 only. Several genes related to LC3-associated phagocytosis, autophagy, or both were differentially expressed. Patients with CAPA had significantly lower neutrophil cell fractions than did patients with COVID-19 only. Patients with IAPA or CAPA had downregulated IFNγ signalling compared with patients with influenza only or COVID-19 only, respectively. The concentrations of several fibrosis-related growth factors were significantly elevated in the bronchoalveolar lavage fluid from patients with IAPA versus influenza only and from patients with CAPA versus COVID-19 only. In one patient with CAPA, we visualised an active or very recent SARS-CoV-2 infection disrupting the epithelial barrier, facilitating tissue-invasive aspergillosis.InterpretationOur results reveal a three-level breach in antifungal immunity in IAPA and CAPA, affecting the integrity of the epithelial barrier, the capacity to phagocytise and kill Aspergillus spores, and the ability to destroy Aspergillus hyphae, which is mainly mediated by neutrophils. The potential of adjuvant IFNγ in the treatment of IAPA and CAPA should be investigated.FundingResearch Foundation Flanders, Coronafonds, the Max Planck Society, the Fundação para a Ciência e a Tecnologia, the European Regional Development Fund, “la Caixa” Foundation, and Horizon 2020.

Pulmonary and central nervous system aspergillosis in a patient with COVID-19 infection

Introduction. Patients with COVID-19 infection are vulnerable to a variety of serious complications, including invasive fungal infections such as aspergillosis. Because pulmonary aspergillosis is difficult to confirm with perfect confidence, it has been classified as ?proven,??probable,? and ?possible.? We present a patient with COVID-19 infection in whom a ?probable? pulmonary aspergillosis was complicated by hematogenous spread into brain with formation of multiple abscesses. Case outline. A 67-year-old female was diagnosed with COVID-19 infection using polymerase chain reaction (PCR) from a nasopharyngeal swab. The patient had never been vaccinated before. Despite standard therapy and noninvasive oxygen support, the patient?s health deteriorated one month following the onset of the disease, with chest discomfort, cough, and hemoptysis. Thoracic computed tomography (CT) revealed bilateral infiltrative lesions with varying diameters of cavities, primarily in the left lung, as well as modest effusions in both pleural spaces. Aspergillus hyphae were isolated from tracheobronchial aspirates. Despite therapy with Amphotericin B, which was only available antifungal medication at the time, the patient fell into a coma. A CT scan of the skull revealed several infiltrative lesions inside the brain, some with cavities suggestive to metastatic abscesses, most likely of fungal etiology (Aspergillus) as a result of hematogenous spread of pulmonary aspergillosis. Despite therapy and all other precautions, the patient died. The autopsy was not carried out. Conclusion. In addition to other complications, COVID-19 patients may develop pulmonary aspergillosis, which can be fatal because of the possibility of hematogenous spread to the brain.

Rare presentation of Aspergillus as cystic swelling in pre-maxillary region

<p class="abstract">Subcutaneous fungal infections are caused by penetration of the causative fungi into the subcutaneous layer and usually present as single, localised non-tender nodular swelling. The diagnostic process is a vital dynamic process that requires effective communication and efficient collaboration. Aspergillus hyphae invade host tissues through release of various toxins like proteases, phospholipases, hemolysins, gliotoxin, aflatoxin, phthioic acid and other toxins. Under general anesthesia and proper aseptic conditions using Moure incision surgical debulking of the pre-maxillary mass was done and post-operatively oral antifungal medication was started. Multiple cystic swelling was sent for histopathological examination and found to be non-invasive Aspergillus fungal infection. Subcutaneous form of aspergillosis manifest as subcutaneous fungal infection. We presented an unusual case report of 45 years old immunocompetent female with cystic presentation of aspergillosis involving premaxillary region.</p>

Selection and characterization of two monoclonal antibodies specific for the Aspergillus flavus major antigenic cell wall protein Aflmp1

Aspergillus flavus is a major fungal pathogen of plants and an opportunistic pathogen of humans. In addition to the direct impact of infection, it produces immunosuppressive and carcinogenic aflatoxins. The early detection of A. flavus is therefore necessary to diagnose and monitor fungal infection, to prevent aflatoxin contamination of food and feed, and for effective antifungal therapy. Aspergillus-specific monoclonal antibodies (mAbs) are promising as diagnostic and therapeutic reagents for the tracking and treatment of Aspergillus infections, respectively. However, A. flavus has a complex cell wall composition and dynamic morphology, hindering the discovery of mAbs with well-characterized targets. Here we describe the generation and detailed characterization of mAb5.52 (IgG2aκ) and mAb17.15 (IgG1κ), which bind specifically to the highly immunogenic cell wall antigen A. flavus mannoprotein 1 (Aflmp1). Both mAbs were generated using hybridoma technology following the immunization of mice with a recombinant truncated version of Aflmp1 (ExD, including the homologous CR4 domain) produced in bacteria. We show that mAb5.52 and mAb17.15 bind specifically to A. flavus and A. parasiticus cell wall fragments (CWFs), with no cross-reaction to CWFs from other fungal pathogens. Immunofluorescence microscopy revealed that both mAbs bind to the surface of Aspergillus hyphae and that mAb17.15 also binds to spores. The epitope for both mAbs is localized within the CR4 region of the Aflmp1 protein. These Aspergillus-specific mAbs may be useful for the early detection of fungal infection in food/feed crops, for serodiagnosis in patients with invasive aspergillosis caused by A. flavus infection and for the development of antibody-expressing disease-resistant crops.

A highly sensitive and specific method to evaluate viable fungal burden of Aspergillus fumigatus in mice by RT-qPCR for 18S ribosomal RNA

Potent fungicidal activity is one of the key factors of antifungals to overcome invasive pulmonary aspergillosis (IPA). To date, quantification of Aspergillus DNA in the lungs and galactomannan (GM) in serum or bronchoalveolar lavage fluid have been developed as general methods for measuring fungal burden in IPA animal models. However, GM quantification is not supposed to be a suitable method for precise evaluation of the fungicidal effects of antifungals, because killed Aspergillus hyphae can release GM for a certain period until they are eliminated by the host. Therefore, in terms of detecting viable fungal burden of Aspergillus, quantification of Aspergillus DNA has been thought to be a suitable method. Here, to obtain a method with much higher sensitivity, we applied reverse transcription quantitative PCR (RT-qPCR) for A. fumigatus 18S ribosomal RNA to measure the viable fungal burden in murine IPA models. Prior to in vivo tests, we confirmed that the sensitivity of 18S rRNA was nearly 50-fold higher than that of 18S ribosomal DNA in vitro. This highly sensitive method made it possible to evaluate the fungicidal effects of antifungals in a low-inoculation murine IPA model. In this model, single administrations of higher doses of voriconazole and posaconazole, which have fungicidal activity, were able to display fungicidal effects with ≥1 log10 reductions by 18S rRNA quantification, whereas significant reductions in serum GM were not observed. These results suggest that 18S rRNA quantification is a powerful tool for screening novel antifungals with potent fungicidal activity only after a single administration.

Effectiveness of factitious foods and artificial substrate in mass rearing and conservation of Neoseiulus californicus (Acari: Phytoseiidae)

ABSTRACT The use of factitious food instead of natural prey, and artificial substrates instead of leaf-based, could be important steps towards reducing natural enemies̓ rearing costs. Nutritional values of four factitious foods comprising Ephestia eggs, decapsulated Artemia cysts, mould mite immature individuals, and Aspergillus hyphae as alternative food sources for Neoseiulus californicus (McGregor) were determined under laboratory conditions. In addition, suitability of artificial (plastic sheet) and leaf-based substrates for rearing of the predator was assessed. The duration of egg to adult developmental time was shorter when the predator fed on the mould mite compared with the other diets. Artemia cysts and Aspergillus hyphae were not suitable diets. The individuals reared on the mould mite had the highest fecundity and intrinsic rate of increase (r). Due to weak mouthpart of the immature predator in feeding of factitious foods, most of them died in immature stages, or their fecundity in adult stage was low. In a separate experiment, almond pollen was also used in addition to factitious foods for rearing of immature stages of the predator. The adult stage of these individuals had the ability to feed and oviposit on the factitious foods, and female longevity and fecundity of these individuals were higher on E. kuehniella egg (and after that on mould mite) than the others. In addition, the results indicated that the rearing performance of the predator fed on the same diet was higher on the artificial substrate than the leaf-based one. The potential of alternative foods and artificial substrates for mass production and conservation of N. californicus is a suitable and cheap method.

Sinobronchial allergic mycosis syndrome in an elderly male

BackgroundAllergic bronchopulmonary aspergillosis (ABPA) and allergic fungal rhinosinusitis (AFRS) are characterized by hyper-responsiveness of the respiratory tract and the nasal cavity and paranasal sinuses, respectively to Aspergillus species and AFRS causes chronic rhinosinusitis. Herein, we report the first case of sinobronchial allergic mycosis (SAM) syndrome, defined as ABPA with concomitant AFRS, caused by Aspergillus fumigatus patient > 80 years.Case presentationAn 82-year-old male with interstitial pneumonia who returned for follow-up exhibited high-attenuation mucus plug in the right intermediate bronchial trunk, infiltration in the right lung field, and right pleural effusion on regular chest computed tomography (CT). We found unilateral central bronchiectasis in the right upper lobe. Similarly, CT scan of the paranasal sinuses revealed high-attenuation mucus plugs in left ethmoid sinuses. Biopsy specimens from the plugs in the right intermediate bronchial trunk and the left ethmoid sinuses revealed allergic mucin with layers of mucus eosinophils, eosinophil-predominant mixed inflammatory cell infiltrate and Aspergillus hyphae. The patient fulfilled all the major criteria for ABPA and AFRS, and was diagnosed with SAM syndrome. CT scan of the lung and paranasal sinuses revealed apparent amelioration after oral steroid therapy.ConclusionDespite mostly reported in relatively young patients, SAM syndrome can occur in elderly individuals as well.

Challenging Alveolar Hemorrhage Complicating Pneumonia After Liver Transplantation: A Case Report

Alveolar hemorrhage is a life-threatening clinical syndrome often initially thought to be atypical pneumonia. Association with hematopoietic stem cell transplantation is well studied, but not with solid organ transplantation. We report a case of a 54-year-old woman presented with fever and shortness of breath on the third posttransplant day after deceased donor liver transplantation. Imaging studies showed diffuse bilateral pulmonary infiltrates and a positive sequential bronchoalveolar lavage test was revealed during bronchoscopy. Cytomegalovirus antigenemia was present in 8/200,000 white blood cells; Aspergillus galactomannan and Pneumocystis jirovecii were also present. However, only Aspergillus hyphae were found in the sputum culture. Management strategy aimed to treat underlying infections, provide adequate respiratory support, and control inflammation. We proposed that diffuse alveolar hemorrhage should be considered as differential diagnosis in early pulmonary complications after liver transplantation. Early diagnosis and aggressive treatment protocol is the key for a good outcome.

Outbreak of aspergillosis in a flock of geese in Zaria, Nigeria

A goose from a flock of twenty five geese, with history of gaping, sternal recumbency, greenish watery diarrhea and inappetence was presented to the Avian and Poultry Health Unit of Veterinary Teaching Hospital, Ahmadu Bello University, Zaria. Ruffled feathers, drooping of the wings, rales, greenish vent and clear greenish diarrhea were observed on physical examination. At necropsy, congested carcass, enlarged and congested liver and spleen, severely hemorrhagic, mucoid and congested trachea, severely congested lungs with multiple and diffused nodular growth all over the lungs were observed. There was a velvety greenish area in the lungs with black spots at the center and nodular growths on the intercostal muscles. Microscopically, the portions of lungs with nodules were composed of necrotic center with intralesional hyphae and conidia typical of Aspergillus spp., a peripheral inflammatory cell response composed of mononuclear cells infiltration and obliteration of alveolar cells. The mycologic culture allowed the isolation and identification of Aspergillus flavus (A. flavus) from lung samples. The gross and microscopic lesions, in combination with the mycologic identification, provided the diagnosis of pulmonary aspergillosis due to A. flavus infection. CuSO4 at 1 g per 5 liters of drinking water was used for a period of 7 days with no signs of the infection.Keywords: Aspergillus spp., Geese, Gaping, Intralesional hyphae, Zaria

Aspergillosis of Gastrointestinal Tract Mimicking Malignancy: A Case Report

A variety of infectious diseases can have clinical, radiological and histopathological findings that mimic various malignancies. We report a case of a 53 year old immunocompetent man admitted initially as a case of perforation peritonitis with strong clinical suspicion of a gastric malignancy. Histologically, the biopsy from the stomach perforation site showed necrotic gastric wall with scattered atypical cells in muscularis. Initially suspected to be neoplastic, these cells could not be characterized definitively by an immunohistochemistry panel. However, special stains for fungi done on account of necrosis, demonstrated presence of numerous fungal hyphae of Aspergillus in the gastric ulcer slough. Systemic antifungal treatment was instituted after which the patient showed significant clinical improvement and was discharged from the hospital following an uneventful recovery. To the best of the author’s knowledge, this is the first report of a case of Aspergillus infection mimicking gastric malignancy in a non-immunocompromised individual from India.

Cerebral aspergillosis: a retrospective analysis of eight cases

Purpose: Aspergillosis of the central nervous system is very rare. However with recent increases in the use of immunosuppressive agents and antibiotics, its incidence is increasing. We evaluated the demographics, clinical manifestations, laboratory findings, diagnosis, underlying conditions, treatment regimens and outcomes of patients with cerebral aspergillosis (CA). Methods: We retrospectively reviewed data from eight patients with CA hospitalized at a Chinese general hospital from 1 January 2005 to 30 September 2015. Results: Common clinical manifestations included headache and cranial nerve involvement. Four patients underwent biopsy and were pathologically diagnosed with Aspergillus hyphae. One patient was proved to have Aspergillus infection via autopsy. One patient had positive cerebrospinal fluid fungal cultures. The lesion locations were: the cavernous sinus (n = 5, 62.5%), frontal lobe (n = 1, 12.5%), temporosphenoid lobe (n = 1, 12.5%) and cerebellum (n = 1, 12.5%). At the end of follow-up, three patients were cured and five patients had died (mortality rate, 62.5%). Conclusions: Most patients with CA had no significant immunosuppression-related conditions in our study. Aspergillus spp. can infect the central nervous system through several pathways and CA has an atypical clinical manifestation. The use of local tissue puncture, surgery or other invasive means to obtain diseased tissue containing higher levels of Aspergillus, followed by culture or histological examination, can contribute to an early diagnosis of CA and timely therapeutic intervention. The prognosis of CA is poor, but early and adequate use of antifungal drugs with high transfer across the blood–brain barrier and radical surgery to remove lesions can improve the survival rate.

Human Neutrophils Use Different Mechanisms To Kill Aspergillus fumigatus Conidia and Hyphae: Evidence from Phagocyte Defects.

Neutrophils are known to play a pivotal role in the host defense against Aspergillus infections. This is illustrated by the prevalence of Aspergillus infections in patients with neutropenia or phagocyte functional defects, such as chronic granulomatous disease. However, the mechanisms by which human neutrophils recognize and kill Aspergillus are poorly understood. In this work, we have studied in detail which neutrophil functions, including neutrophil extracellular trap (NET) formation, are involved in the killing of Aspergillus fumigatus conidia and hyphae, using neutrophils from patients with well-defined genetic immunodeficiencies. Recognition of conidia involves integrin CD11b/CD18 (and not dectin-1), which triggers a PI3K-dependent nonoxidative intracellular mechanism of killing. When the conidia escape from early killing and germinate, the extracellular destruction of the Aspergillus hyphae needs opsonization by Abs and involves predominantly recognition via Fcγ receptors, signaling via Syk, PI3K, and protein kinase C to trigger the production of toxic reactive oxygen metabolites by the NADPH oxidase and myeloperoxidase. A. fumigatus induces NET formation; however, NETs did not contribute to A. fumigatus killing. Thus, our findings reveal distinct killing mechanisms of Aspergillus conidia and hyphae by human neutrophils, leading to a comprehensive insight in the innate antifungal response.